Influence of Pyrolysis

Conditions on
Char Properties
Bob Hawkins
Managing Research Chemist
Eprida
Athens, GA
 Intro

Char’s ability to provide nutrients

Char’s ability to facilitate nutrient
retention/uptake
Low temperature char –
temperature range 375-500o
Sample Preparation
Batch unit
Sample Preparation
Pilot Plant
 Volatile Content
VM%

28.50%
PN
27.50%
no steam
26.50% pine
25.50% higher temp
no vent
24.50%
vent
23.50%
0 2 4 6 8
psig
 Fixed Carbon
FC%

70.00%
69.00% press
no steam
68.00%
pine
67.00%
higher temp
66.00%
no vent
65.00% vent
64.00%
0 2 4 6 8
psig
 % C

55.0
60.0
65.0
70.0
75.0
80.0
PN
37
1
PN
40
2
PN
42
6
SD
37
8
SD
39
9
SD
41
8
PC
37
9
PC
40
1
PC
42
6
Carbon content

PB
38
2
PB
Charcoal sample ID
39
9
PB
42
Total Nutrients

HW 6
38
HW 2
40
HW 0
42
HR 6
38
HR 7
42
8
 % N

0.00
0.50
1.00
1.50
2.00
2.50
PN
37
PN 1
40
PN 2
42
SD 6
37
SD 8
39
SD 9
41
PC 8
37
PC 9
40
PC 1
42
PB 6
Nitrogen content

38
2
PB
Charcoal sample ID
39
9
PB
42
Total Nutrients

HW 6
38
HW 2
40
HW 0
42
HR 6
38
HR 7
42
8
Total Nutrients
Al B

Ca Cu

Fe K

Mg Mn

Mo Na

P Pb

Zn
 Available Nutrients
Available Potassium

1400 9000
1200 8000
7000 Pine sawdust
1000
6000 Pine chips
mg/kg

800 5000 Pine bark

600 4000 Oak-sapwood
3000 Oak-heartwood
400
2000 Peanut hull pellets
200 1000
0 0
360.0 380.0 400.0 420.0 440.0
Charring temperature (deg C)
 Available Nutrients
Available Phosphorous

150.0 1000.0
Pine saw dust
800.0
100.0 600.0 Pine chips
m g /k g

50.0 400.0 Pine bark

200.0
Oak-sapw ood
0.0 0.0
360.0 370.0 380.0 390.0 400.0 410.0 420.0 430.0 440.0 Oak-heartw ood
Peanut hull pellets
Charring temperature (deg C)
 % of total P % of total K

0.00%
10.00%
20.00%
30.00%
40.00%
50.00%
0.00%
10.00%
20.00%
30.00%
40.00%
50.00%
60.00%
PN PN
37 37
1 1
PN PN
40 40
2 2
PN PN
42 42
6 6
SD SD
37 37
8 8
SD SD
39 39
9 9
SD SD
41 41
8 8
PC PC
37 37
9 9
PC PC
40 40
1 1
PC PC
42 42
6 6
PB PB
38 38
2
Available K as % of total

2
Available P as % of total

PB PB
39 39
9 9
PB PB
42 42
6 6
H
W HW
38 38
2 2
HW HW
40 40
0 0
Available Nutrients

H H
W W
42 42
6 6
H
R HR
38 38
7 7
HR H
42 R4
8 28
 Char pH
Char pH

12

11 Peanut hull pellet s

10
Pine sawdust

9
Pine chips
pH

8
Pine bark

7
Oak-sapwood

6
Oak-heart wood

5
360.00 370.00 380.00 390.00 400.00 410.00 420.00 430.00 440.00
Tem p
 CEC
CEC

19.0
18.0
17.0
PN
16.0
15.0 +250-420
14.0 +500-850
13.0 PN-pilot
12.0
No Steam
11.0
10.0
9.0
350 400 450 500
 CEC
CEC vs Pressure

14.0
12.0
10.0 PN
8.0 No Steam
6.0 Higher temp

4.0 Pine

2.0
0.0
0 2 4 6 8
 Surface Acid Concentration
Charcoal surface acids (Titration w ith NaOH)

2.5
Total amount of surface acids (meq/g)

pine chips
2 Pine saw dust
Pine Bark
Hardw ood (red)
1.5
Hardw ood (w hite)
Peanut hull pellets
1 No Steam
Pilot plant

0.5

0
300 350 400 450 500
Charcoal tem perature
 Char Production

Need for quick test for QA/QC

Use of steam as a sweep gas
shows increases in CEC and
surface acid formation
 Fuel Vol. 75, No. 9, pp. 1051-1059, 1996
Copyright 0 1996 Elsevier Science Ltd
Printed in Great Britain. All rights reserved
ELSEVIER PII:SOO16-2361(!Xi)OOO81-6 0016-2361/96 %15.00+0.00

Influence of temperature on the products

from the flash pyrolysis of biomass

Patrick A. Horne and Paul T. Williams

Department of Fuel and Energy, The University of Leeds, Leeds LS2 9JT, UK
(Received 12 May 1995; revised 27 March 1996)

Biomass in the form of mixed wood waste was pyrolysed in a fluidized bed reactor at 400, 450, 500 and
550°C. The char, liquid and gas products were analysed to determine their elemental composition
and calorific value. In particular, the liquid products were analysed in detail to determine the concentration
of environmentally hazardous polycyclic aromatic hydrocarbons (PAH) and potentially high-value
oxygenated aromatic compounds in relation to the process conditions. The gases evolved were COZ,CO and
C,-C4 hydrocarbons. The liquids were homogeneous, of low viscosity and highly oxygenated.
The molecular weight range of the liquids was 50-1300~. Chemical fractionation of the liquids showed
that only low quantities of hydrocarbons were present and the oxygenated and polar fractions were
dominant. PAH up to MW 252 were present in the liquids; some of the PAH identified have been shown to
be carcinogenic and/or mutagenic. The concentration of PAH in the liquids increased with pyrolysis
temperature, but even at the maximum pyrolysis temperature of 550°C the total concentration was
< 120ppmw. The liquids contained significant quantities of phenolic compounds and the yield of phenol
and its alkylated derivatives was highest at 500 and 550°C. Some of the oxygenated compounds identified
are of high value. Copyright 0 1996 Elsevier Science Ltd.

(Keywords: biomass; flash pyrolysis; products)

The energy potential of biomass and solid wastes has
become increasingly recognized as a means to help meet
world energy demand. The use of biomass has a
particularly important role as an energy source in
developing countries1’2. The utilization of biomass and
other alternative fuel sources rather than existing fossil
fuels could offer more environmentally acceptable
processes for energy production and will aid in conser-
ving the limited supplies of fossil fuels. The recovery
of energy from biomass and solid wastes has centred
on biochemical and thermochemical processes3. Of
the thermochemical processes, pyrolysis has received
increased interest, since the process conditions can be
optimized to maximize the production of chars, liquids
or gasesle3. In particular, the production of pyrolysis
liquids has been investigated with the aim of using the
liquid product directly in fuel applications or by
producing refined fuels and/or chemical
;;t;zEfj . The solid char can be used as a fuel in the
form of briquettes or as a char-oil/water slurry, or it can
be upgraded to activated carbon and used in purification
processes”*. The gases generated have a low to medium
heating value but may contain sufficient energy to supply
the energy requirements of a pyrolysis plant.
The physical conditions of the pyrolysis of biomass,
such as temperature, heating rate and residence time,
have been shown to have a profound effect on the
product ields and composition . High heating rates of
up to lc& s-1 at temperatures < 650°C and with rapid
quenching, fav&r the formation of liquid products and
minimize char and gas formation; these process conditions
are often referred to as ‘flash pyrolysis’. High heating
rates to temperatures > 650°C tend to favour the
formation of gaseous products at the expense of liquids.
Slow heating rates coupled with low maximum tempera-
tures maximize the yield of char.
In recent years, various flash pyrolysis processes have
been developed to maximize the formation of liquid
products for use as fuels or chemical feedstocks. A vortex
reactor process has been developed by Diebold”“. This
type of pyrolKsk process is an ablative technique. Scott
and Piskorz ’ have designed the Waterloo flash
pyrolysis process to maximize the liquid product yield.
They have used small biomass particles (< 1 mm) as the
feed and high heat transfer rates with a hot fluidized bed
of sand. Georgia Tech in the USA has used an entrained
flow pyrolysis reactor where the biomass particles are
rapidly heated by a flow of hot gases13. Vacuum pyrolysis
has also been used as a flash pyrolysis technique by Roy
et a1.14. The liquid product yields from all the above
processes were >50 wt%, with Dieboldgl” and Scott and
Piskorz11112reporting yields of >70 wt%.
The liquid products from flash pyrolysis processes
have been reported as being homogeneous and of
low viscosity, and are chemically extremely complex,
containing hundreds of different components’5-‘7. The
chemical composition of the liquid hydrocarbons and
the relation of composition to process conditions has
implications for end use as a fuel or chemical feedstock.
Many biomass-derived pyrolysis oils are known to
contain polycyclic aromatic hydrocarbons (PAH)18-20,
some of which have been shown to be carcinogenic and/

Fuel 1996 Volume 75 Number 9 1051

Influence of temperature on flash pyrolysis of biomass: P. A. Horne and P. T. Williams
n S7ktyWVe
II
4-
Figure 1 Schematic diagram of the fluidized bed pyrolysis reactor
or mutagenic*l’**, which may have consequences for
the handling of the fuel. Where the proposed end use of
the liquid hydrocarbons is as a chemical feedstock, again
the process conditions which optimize the formation of
high-value chemicals in the liquid have economic
benefits. In this context the oxygenated aromatic
hydrocarbons are of particular interest. Biomass-derived
oils have been shown to contain phenols’5’23 which have
extensive use in the production of resins. In addition,
phenolic derivatives have a high value, as they are used as
flavourings in the food industry. Syringol and guaiacol
are also found in significant concentrations in biomass-
derived pyrolysis oils and are used in the production of
biodegradable polyesters and polyethers.
In this work, a semi-continuous fluidized bed reactor
was used to flash-pyrolyse a waste wood feed. The
pyrolysis temperature range studied was 400-550°C as
this is known to give high yields of liquid products whilst
minimizing the formation of char and gases. Pyrolytic
liquids derived from biomass have been analysed by a
number of workers15-17, but there are limited data on the
individual yields of PAH and oxygenated aromatic
compounds in relation to process conditions, coupled
with analyses of the gas phase and char. Therefore in this
Table 1 Proximate and ultimate analyses (wt%) of the wood feed
Proximate analysis
Volatiles 91.0
Moisture 1.5
Ash 1.5
Ultimate analysis
C 45.9
H 5.12
0 46.6
1052 Fuel 1996 Volume 75 Number 9
work the pyrolysis liquids were characterized in detail in
relation to process conditions, with particular reference
to the contents of PAH and oxygenated aromatic
compounds.
EXPERIMENTAL
Biomass
The biomass used was a mixture of waste wood
shavings obtained from a woodworking company, and
therefore represented a mixture of different wood types.
Table 2 shows the proximate and ultimate analyses of the
biomass pyrolysed.
Pyrolysis reactor
The reactor system used was a fluidized bed pyrolysis
unit (Figure I). The reactor was 7.5 cm diameter x 50 cm
high, constructed from stainless steel. The fluidization
gas was nitrogen, preheated before entry into the reactor.
The flow-rate of nitrogen was sufficient to provide three
times the minimum fluidizing velocity (MFV) to the bed.
The bed material was quartz sand with a mean diameter
of 250 pm and a static bed depth of 8 cm. The biomass
was fed to the reactor via a screw feeder and nitrogen gas
stream to the top of the fluidized bed at a rate of 0.216-
0.228 kg h-l. The residence time of the pyrolytic vapours
in the hot reactor was N 2.5 s at a pyrolysis temperature
of 500°C. The pyrolysis vapours leaving the reactor were
passed through a series of condensers. The initial
condensation was provided by two stainless steel
condensers which were water-cooled both internally
and externally, with the catch pots for each condenser
ice-cooled. There followed a series of glass condensers
cooled using a mixture of solid carbon dioxide and
acetone, which were used to remove any residual vapours
from the gas stream. This condensation system was
found to be 97% efficient in trapping volatile aromatic
hydrocarbons such as toluene and 99% efficient for the
condensation of water. The pyrolysis condensate from all
the condensers was mixed and stored at -10°C. The
water fraction of the pyrolysis liquids was separated
from the organic fraction using the standard ASTM
D244 and IP 29.1 methods.
Gas analysis
The carrier gas stream was sampled after the
condensation system to allow for the analysis of any
non-condensable gases. Analysis was carried out using
gas chromatography. The gases determined were CO and
H2 using a molecular sieve SA 60-80 column and CO2
using a silica column, with argon as carrier gas and
thermal conductivity detection. For the determination of
hydrocarbon gases up to Cs a Porisil C 80-100 column
was used, with nitrogen as carrier gas and a flame
ionization detector.
Elemental analysis
The carbon and hydrogen contents of the pyrolysis
liquids and char were determined using an elemental
analyser. The oxygen content of the liquids and chars
was found by difference.
Calor@ic value
The calorific value of the pyrolysis liquid was
 Influence of temperature on flash pyrolysis of biomass: P. A. Horne and P. J. Williams

determined by bomb calorimetry. The values reported
are the gross heat of combustion at constant volume.
Oil analysis
Molecular weight range. The molecular weight (MW)
range of the oils was determined using a mini-column
size exclusion chromatography (s.e.c.) system which has
been described previouslg4. The system incorporated two
150 mm x 4.6 mm i.d. columns with Polymer Labora-
tories 5 pm RPSEC 100 A type packing. A third column
of the same material was placed in line between the pump
and the injection valve, to ensure pre-saturation of the
solvent with the column packing material and also to
avoid analytical column dissolution and hence loss of
performance. The solvent used for the mobile phase
was tetrahydrofuran (THF), which has been shown by
Johnson and Chum25 to be suitable for the analysis of
biomass pyrolysis oils. The calibration system used was
based on polystyrene samples of low polydispersity in
the MW range from 800 to 860000; also included was
benzene for low-MW calibration. Samples were intro-
duced through a 2 ,ul loop injection valve. Two detectors
were used: a U.V. detector sensitive to the aromatic
compounds, and a refractive index (RI) detector moni-
toring the elution of all compounds, enabling more infor-
mation to be obtained regarding the relative contribution
to the MW of the difference chemical class fractions24.
Ultraviolet scanning of the polystyrene MW fractions
in THF indicated that the maximum absorbance was at
262nm. Determination of the maximum efficiency for
the system showed that a mobile phase flow rate of
0.26 mL min-’ and a column temperature between 2
and 14°C were optimum24. For practical purposes the
column was maintained at 0°C. Column temperature is
usually maintained at room26 or elevated
temperature27-29. However, the optimum efficiency was
obtained at lower than ambient temperatures for the
system used in this work.
Evaluation of the system has shown that there are
systematic variations in the measured MW of n-alkanes,
n-alkenes, PAH and alkylated cyclic hydrocarbons
compared with the polystyrene standards24. Deviations
from polystyrene calibration curves have also been
shown previously for other model compounds26>30>31.
For biomass pyrolysis oils, Johnson and Chum25 used
polystyrene MW fractions and showed that aromatic
acids and naphthalenes deviated significantly from the
calibration curve. However, in this work the s.e.c. system
was used to compare oils derived from different process
conditions rather than to determine the absolute MW.
The fractions were analysed by Fourier transform
infrared spectroscopy (FT-i.r.) to determine the efficient
separation of the chemical classes. In addition, each
fraction was analysed by gas chromatography-mass
spectrometry using ion trap detection (g.c.-ITD) to
verify the fractionation scheme. The percentage mass in
each class fraction was determined by analysis of the
total gas chromatogram in relation to known masses of
sample and standards analysed by the system. This
avoided the necessity to nitrogen-evaporate (blow
down) the solvent eluant, which may result in losses of
light hydrocarbons.
Detailed characterization of the pyrolysis liquids. The
concentrations of monocyclic and polycyclic aromatic
hydrocarbons (PAH) in the benzene fractions and the
oxygenated aromatic species present in the ethyl acetate
fraction were determined using gas chromatography-
mass spectrometry (g.c.-m.s.) The g.c.-m.s. system
was a Carlo-Erba Vega HRGC with cold on-column
injections, coupled to a Finnigan Mat ion trap detector
(ITD). A DB-5 fused silica capillary column 25mx
0.3mm was used and the temperature programme was
60°C for 2min followed by 5 Kmin-’ to 270°C with a
dwell time of 25 min at 270°C. The ITD mass range
was set at 30-350~ with a scan time of 0.125-2 s.
The ITD was linked to a PC with a mass spectral library
facility. Identification of the individual constituents in
the chemical fractions was carried out using the g.c.-
m.s. and retention indices23,32Y33. Single-ion monitoring
(SIM) was also carried out to confirm the identification
of compounds and also to examine the samples for a
series of substituted compounds, for example, naphthalene
and methyl-, dimethyl- and trimethylnaphthalenes.
Quantification was determined by the use of extensive
external standards.
RESULTS AND DISCUSSION
The yields of the pyrolysis products are shown in Table 2.
Several repeat runs were carried out at 550°C under
identical conditions to test the repeatability of the
process. The differences observed between the yields of
the char, liquid and gas for these repeat runs were
negligible. The liquid yield for all the runs was >65 wt%,
with a maximum yield of 67.8 wt% at 550°C. At
pyrolysis temperatures of 500 and 550°C the pyrolysis
Table 2 Product yields (wt%) from the flash pyrolysis of wood

Reactor temperature (“C) Char Liquid Gases Total

Chemicalfractionation. The biomass-derived pyrolysis
oils were fractionated using mini-column liquid chroma- 400 24.1 65.5 10.2 99.8
450 21.4 65.7 11.1 98.2
tography. The mini-columns were conditioned by washing 500 18.9 66.0 14.6 99.5
with n-pentane. A 250 mg sample of the oil was placed on 550 17.3 67.0 14.9 99.2
the column. The samples were added by adsorption on to 550 16.7 67.8 15.7 99.2
inert Chromosorb G/AW/DMCS 60-80 support, mixed 550 17.1 66.2 15.2 98.5
and then packed above the silica section of the column.
This approach is necessary for polar oils, which may Table 3 Water and organic contents (wt%) of the liquid product from
produce a solid-phase precipitate with the n-pentane the flash pyrolysis of wood
solvent and block the column, and also to improve
solvent contact with the oil. The column was then eluted Reactor temperature (“C) Water Organic material
with n-pentane, benzene, ethyl acetate and methanol 400 28.0 72.0
(polarity relative to Al2O3, 0.00, 0.32, 0.58 and 0.95 450 27.6 72.4
respectively), to produce aliphatic, aromatic, oxygenated- 500 29.6 70.4
550 26.8 73.2
aromatic and polar chemical class fractions respectively.

Fuel 1996 Volume 75 Number 9 1053

Influence of temperature on flash pyrolysis of biomass: P. A. Horne and P. T. Williams

liquid was a homogeneous dark liquid of low viscosity. temperatures. However, the secondary decomposition
Similar observations were made by Scott and Piskorz ’ of the char at higher temperatures may also give
using a fluidized bed process with a pyrolysis tempera- non-condensable gaseous products. Beaumont and
ture of 500°C. The pyrolytic product from the 400 and Schwob34 and Samolada et aL3’ investigated the flash
450°C fluidized bed pyrolysis temperature experiments pyrolysis of wood and found that as the pyrolysis
was found to be of low viscosity but there were traces of temperature was raised, the gaseous yield was increased.
a black tar residue on the base of the storage vial. The The yield of CO and CO2 in general increased with
pyrolysis liquid product was a mixture of organic increasing pyrolysis temperature. However, the increase
material and water. The water content of the liquid in CO was far more pronounced than that of C02. The
was determined using the ASTM D244, IP29.1 method; yield of all the hydrocarbon gases increased with
Table 3 shows the results of this separation. The water temperature. Similar observations have been made by
content of all the pyrolytic liquids was approximately other workers11’35.
the same (26.8-29.6wt%). The initial wood feed
contained 7.5 wt% moisture, which would be released Elemental analysis of pyrolysis products
during pyrolysis and subsequently collected during The liquid and char products were analysed to
condensation. This contributed approximately half determine their elemental composition. Table 5 shows
the water present in the pyrolytic liquid. Therefore the elemental compositions of the wet pyrolytic liquid,
water formed by pyrolysis accounts for 14 wt% of the the oil after removal of water, and the chars. The
liquid condensate or lOwt% of the initial wood feed. elemental compositions of the pyrolytic liquids both
Table 3 also shows that over the temperature range before and after removal of water were similar for all the
studied, the yield of oil was not significantly affected by a pyrolysis temperatures investigated. The similarity was
change in temperature. probably due to the particular temperature range
The char yield was reduced as the pyrolysis temperature studied, the lowest temperature of 400°C being adequate
was increased, from 24.1 wt% at 400°C to 16.7 wt% at to decompose the wood feed, and the low residence
550°C. The char yields reported in this work were higher time limiting the amount of secondary reactions. The
than those found by other workers1”12. This difference pyrolysis liquids arising from the reactor before water
was probably due to the fact that a mixed wood waste removal had a carbon content lower than that of the
was used in this work rather than a single known biomass initial wood feed. This indicates that such pyrolytic
feedstock as used by Scott et al.11>‘2.The decrease in the liquids have a low CV, and the removal of the water is
char yield with increasing temperature could be due necessary to maximize their CV. The composition of the
either to greater primary decomposition of the wood at pyrolytic liquid produced in this work compares well
higher temperatures or to secondary decomposition of with the finding of Churin36.
the char residue.
The gaseous product yield increased with pyrolysis Characterization of the pyrolysis liquids
temperature. The individual yields of the major gaseous Molecular weight range. The pyrolytic liquid after
species are shown in Table 4. The increase in gaseous water removal was used for determination of the MW
products is thought to be predominantly due to range of the oils, because the presence of water caused
secondary cracking of the pyrolysis vapours at higher the RI detector to give inconsistent results. Figure 2
shows MW ranges of the pyrolysis oils for the RI and
U.V. detectors. Only the data for the 400 and 550°C
Table 4 Yields (wt%) of gases from the flash pyrolysis of wood pyrolysis oils are shown, for clarity. The oils all showed
Reactor teme.
similar MW ranges from 50 to 1300~ for both the RI
(“(7 L CO COP Hz CH4 C2H6 W4 C3h C3H6
and U.V. detectors. The similarity in MW among the
oils over the temperature range studied was due to the
400 3.75 6.02 0.018 0.21 0.05 0.05 0.03 0.05 fact that the lowest temperature, 4OO”C, was sufficient
450 4.20 6.32 0.022 0.35 0.05 0.08 0.02 0.08
500 6.76 6.61 0.022 0.58 0.09 0.26 0.05 0.19 to give almost complete decomposition of the wood
550 6.71 6.86 0.023 0.69 0.16 0.26 0.04 0.45 feed and the low residence times minimized secondary
reactions which might have occurred at higher tempera-
tures. The average MW was -220 and ~275 u for the RI
Table 5 Elemental composition (wt%) of the products from the flash and U.V.detectors respectively. Diebold et a1.37 obtained
nvrolvsis of wood
flash pyrolysis oils from a vortex reactor system at
Reactor temp. (C) C H 0 625°C. They carried out s.e.c. analysis of the oil and
found components present in the oil up to and including
Total liquid product
400 38.6 8.52 51.7
2000~. Evans and Milne2’ used molecular beam mass
450 39.9 8.61 50.4 spectrometry (MB-m.s.) to analyse the pyrolysis vapours
500 37.6 8.42 53.0 whilst in the pyrolysis reactor. They found molecular
550 38.1 8.46 52.8
Liquid product after water removal
400 58.1 6.10 34.6
450 58.0 6.24 35.0 Table 6 Chemical fractionation of the pyrolysis liquids (wt%)
500 57.2 6.12 35.2
Reactor Pentane Benzene Ethyl acetate Methanol
550 59.6 6.05 33.5
temp. (“C) eluate eluate eluate eluate Total
Char
400 68.1 3.23 28.2 400 <0.2 0.42 36.2 56.8 94.6
450 71.9 3.16 24.2 450 <0.2 0.51 37.4 57.6 95.7
500 73.0 3.17 22.9 so0 <0.2 0.54 39.3 56.7 96.7
550 71.6 2.65 24.4 550 <0.2 0.54 38.4 55.2 95.3

1054 Fuel 1996 Volume 75 Number 9

 influence of temperature on flash pyrolysis of biomass: P. A. Horne and P. T. Williams

Table 7 Concentrations (ppmw) of the aromatic compounds present

in the pyrolysis oils

Reactor temperature (“C)

400 450 500 550

Benzene 7 14 55 97
Toluene 5 12 35 6-l
Dimethylbenzene 1 1 2 I
Ethylbenzene 2 2 7 24
Dimethylbenzene 1 1 3 12
Trimethylbenzene 5 5 7 9
Dihydroindene <l <l <l 4
Indene 2 3 6 2
Benzofuran 1 2 4 10
0 Methylbenzofuran 3 2 5 13
1.6 1.8 2.0 2.2 2.4 2.6 2.8 3.0 3.2 Tetramethylbenzene <l <l <l <l
Log Molecular Mass Methylindene <l <l <l 2
Naphthalene 3 5 4 11
(b) Methylnaphthalene 3 4 9 14
Biphenyl 1 1 1 2
Acenaphthene <l 1 1 3
Dimethylnaphthalene 3 5 9 15
Trimethylnaphthalene <l 3 5 7
Tetramethylnaphthalene <l <l <l <l
Fluorene <l <l 1 1
Methylfluorene <l <l 1 <l
Phenanthrene 3 5 6 11
Anthracene 2 2 4 6
Dimethylfluorene <l <l <l <l
Methylphenanthrene I I 10 16
Dimethylphenanthrene 3 I 8 12
Trimethylphenanthrene <l 1 1 2
b Tetramethylphenanthrene <l <l <l 2
oi,4,,,,
16 1.6 2.0
, ,
2.2 2.4
‘, ,

2.6
” ,

2.8
,

3.0
), I
3.2
Pyrene
Methylpyrene
<l
<l
<l
<l
<l
<l <l
1

Log MolecularMass Dimethylpyrene <l <l <l <l

Chrysene <l <l <l <l
Figure 2 Molecular mass range of the pyrolysis liquid at 400 and Methylchrysene <l <l <l <l
550°C after removal of water, using refractive index and ultraviolet Benzopyrene <l <l <l <l
detection

The results of the chemical fractionation are shown in

weights predominantly between 100 and 150 u. The dif-
Table 6, corrected to account for the removal of water.
ference between the findings of Evans and Milne and
They show that the liquids are almost exclusively made
the present results is due to the fact that the condensed
up of oxygenated compounds. The ethyl acetate fraction,
pyrolysis liquids were analysed in this work and that of
which contains the phenolic and aromatic oxygenated
Diebold et al.37. It is thought that some of the smaller
compounds, shows a small but significant increase in
species present in the pyrolysis vapours are highly reac-
concentration from 36.2 to 39.3 wt% of the oil as the
tive and on condensation can polymerize to form
pyrolysis temperature increases from 400 to 550°C. The
higher-molecular-weight material’.
polar material present in the methanol fraction of the
Johnson and Chum25 have suggested that the high
liquids shows no significant trend with change in
apparent MW of biomass pyrolysis oils may be due to
temperature, varying from 55.2 to 57.6 wt%. The hydro-
solute-solute or solute-solvent association, producing
carbons, eluted in the pentane and benzene fractions,
high-molecular-weight complexes. In this work, the
account for < 1 wt% of the pyrolytic liquid. The benzene
samples were prepared with THF immediately prior to
fraction, containing aromatic species, shows a small
analysis, since previous work has shown that storage of
increase with increasing pyrolysis temperature. The
the sample in THF for extended periods did result in an
similarity of the results for the chemical fractionation
increase in apparent MW due to interaction of the
of the oils over the temperature range studied is again
sample with the solvent3*.
due to the fact that the fluidized bed process minimized
The U.V.detector was used to obtain detail on the MW
any secondary reactions and the lowest temperature,
range of the aromatic fraction of the pyrolysis oils. It
400°C was sufficient to give almost complete decomposi-
showed that there was some high-molecular-weight
tion of the wood feed.
material present in the oils which was aromatic in
nature. This could be formed from the decomposition of
the lignin fraction of the wood. Characterization of the chemical fractions of the pyrolysis
liquids
Chemical fractionation. The pyrolytic liquids after PAH content. The major aromatic hydrocarbon
removal of water were separated into four chemical compounds present in the pyrolysis oils are shown in
classes by sequential elution of the column with pentane, Table 7, with PAH compounds up to molecular mass
benzene, ethyl acetate and methanol to give aliphatics, 252 identified and quantified. The major aromatic hydro-
aromatics including PAH, phenolic and neutral oxyge- carbons present in the pyrolysis liquids were the
nated compounds, and polar compounds respectively. monocyclic compounds such as benzene, toluene and

Fuel 1996 Volume 75 Number 9 1055

influence of temperature on flash pyrolysis of biomass: P. A. Horne and P. T. Williams
dimethyl- and ethylbenzenes. The PAH found were
naphthalene and phenanthrene and their alkylated
derivatives and in minor concentrations other compounds
such as pyrene, chrysene and benzopyrenes. The concen-
tration of PAH increased with increasing temperature.
The PAH found have been shown to be carcinogenic
and/or mutagenic2113g@. However, even at the highest
temperature of 550°C the total concentration of PAH
quantified was < 120 ppmw.
PAH have been detected by other workers in biomass
pyrolysis oils. For example, Pakdel and Roy41 analysed
oil from the pyrolysis of Aspen poplar wood chips in a
vacuum pyrolysis unit. They found a wide range of PAH
including naphthalene, phenanthrene and fluorene and
their alkylated substituents, in addition to benzene and
its alkylated substituents. In addition, they quantified
certain PAH, some of which were biologically active,
such as benzo[a]pyrene, chrysene and benzo[k]fluor-
anthene, but were present in very low concentration,
<5ppmw. However, the oil was collected at a biomass
pyrolysis temperature of only 263°C. Gasification tars,
produced at higher temperatures (XSO’C) than the
pyrolysis oils, were found to contain much higher
concentrations of PAH. However, the yield of gasification
tar was small relative to that of the biomass pyrolysis
oils. Elliottlg has also confirmed that PAH are not
present in pyrolysis oils produced at temperatures
<5OO”C but markedly increase in concentration in
gasification tars produced >7OO”C. Desbene et ~21.‘~
analysed pyrolysis oils from the slow pyrolysis of
hornbeam biomass. The PAH detected included
alkylated naphthalenes, biphenyls, fluorene, anthracene,
pyrene and benzofluorene. The composition of the oil
depended on whether the pyrolysis was slow or fast. The
low concentrations of PAH in the pyrolysis oils in
the present work are due to the process conditions used.
The relatively low temperatures and residence times
restrict the formation of PAH by reducing the amount of
possible secondary reactions. For significant quantities
of PAH to form during the pyrolysis of biomass, long
residence times are needed at temperatures >700”C20.
Table 7 shows that as the temperature of pyrolysis
was increased, the concentration of PAH in the oils also
increased. For each individual PAH there was an increase
with increasing pyrolysis temperature. The reactions of
the pyrolysis vapours at increased temperatures result in
the formation of PAH. The formation of aromatic and
polyaromatic hydrocarbons by secondary reactions
during pyrolysis has been attributed to at least two
mechanisms: a Diels-Alder type reaction, and deoxy-
genation of oxygenated aromatic compounds42.
The calorific values of the pyrolysis liquid after
removal of water, the char and the ases from the
550°C run were found to be 22MJ kg- g , 25.9 MJ kg-’
and 15.7 MJ me3 respectively. The original wood feed
had a CV of 17.7 MJ kg-‘. Therefore the pyrolysis liquid
after removal of water contained -63% of the potential
energy in the wood feed. The density of the pyrolytic
liquid (1.2 g cme3) was also much greater than that of the
initial feed (0.4 gcmp3), giving a much higher energy
density. There was a significant amount of water present
in the pyrolytic liquid which would significantly reduce
its CV. Indeed, the elemental analysis shows that the
pyrolysis liquid before removal of water contains
significantly less carbon than the original biomass.
However, removal of the water may not be beneficial,
1056 Fuel 1996 Volume 75 Number 9
as the liquid product from the flash pyrolysis of wood has
been shown to be unstable at elevated temperatures and
polymerizes when exposed to air37. The viscosity of the
pyrolytic liquid may also be increased by removing the
water, which could affect its use as a fuel.
There are conflicting opinions as to the potential uses
of pyrolytic liquids derived from biomass. Maggi et aI.43
state that flash pyrolysis bio-oils are corrosive, are not
completely volatile, have a high oxygen content and do
not mix readily with conventional fuels. They state
that bio-oils used in direct combustion processes rarely
meet the standards required for fuels. Rapper questions
whether pyrolysis liquids meet the requirements of a
storable liquid fuel, and states that pyrolysis oils are
much lower in quality than even a heavy fuel oil,
which itself has a rapidly shrinking market. However,
Bridgwater45 has suggested various processes where flash
pyrolysis liquids could be used both now and in the
future. Many of the processes that he suggests require the
pyrolysis liquids to be refined or upgraded before they
are suitable for use. Solantausta et a1.46 used flash
pyrolysis oils as a fuel in a diesel power plant. They
concluded that the preliminary results were positive but
that further research was necessary on the storage of the
oils and that the oils themselves required more detailed
characterization. More detailed characterization of the
pyrolysis oils is necessary to optimize their potential.
The PAH content of any fuel is of great importance, as
these compounds are environmentally hazardous and
therefore their concentration in a fuel should be
minimized. The process conditions used in this work
show that flash pyrolysis gives only low levels of PAH
formation from biomass feedstocks.
Ethyl acetate fraction. The major compounds present
in the ethyl acetate eluates of the pyrolysis liquids were
identified and quantified; the results are shown in Table
8. The major constituents of the eluate appear to be light
organic oxygenates and phenolic material. The major
phenolic compounds present in the ethyl acetate eluate
of the pyrolysis liquid were phenols, benzenediols, meth-
oxyphenols and dimethoxyphenols. The concentration
of phenol and its alkylated derivatives increased as the
pyrolysis temperature was increased, but 500 and
550°C gave similar yields. The formation of methoxyphe-
no1 and its mono- and dimethyl derivatives was greatest
at the lower pyrolysis temperatures. This was also true
for the overall concentration of dimethoxyphenol and
its derivatives. The more severe pyrolysis temperatures
of 500 and 550°C would increase the possibility of
secondary reactions that could be responsible for the
thermal breakdown of the larger phenolic compounds
such as methoxy- and dimethoxyphenols to phenol,
which would then undergo alkylation, thus giving the
increase in alkylated phenols observed at 500 and 550°C.
The ethyl acetate fraction also contained large
amounts of methylmethoxy-, dimethoxy-, hydroxy-
methoxy- and dihydroxymethoxyphenylethanones. The
concentration of these compounds in the pyrolysis
oils varied, depending on the pyrolysis temperature.
The other major group of compounds present in this
fraction comprised cyclopentanone, cyclopentenone and
the methyl and hydroxymethyl derivatives of cyclopen-
tenone. The concentration of these compounds in the
pyrolysis oils increased as the pyrolysis temperature was
increased. There were large amounts of light organic
 Influence of temperature on flash pyrolysis of biomass: P. A. Horne and P. T. Williams

compounds such as glycol aldehyde and the propyl ester
of acetic acid, which were greatest in the pyrolysis oil
produced at 450°C. Other light organics present in the
oils were furanmethanol, furanone and methylfurfural.
The concentration of these compounds in general
increased with pyrolysis temperature.
In the work the biomass pyrolysis liquids have
been shown to contain significant quantities of phenolic
compounds. These phenolic compounds could be
removed from the pyrolysis liquids prior to their
combustion, as they have a significant commercial
value47148. The use of the pyrolysis liquids for the
production of phenolic chemical feedstocks as well as
for the production of liquid fuels would increase their
potential commercial exploitation. In maximizing the
formation of individual phenolic compounds, it must be
taken into account that an increase in pyrolysis
temperature in this work tended to increase the forma-
tion of phenol and its alkylated derivatives whilst
reducing the formation of the larger phenolic
compounds. Stoikos48 has reviewed the upgrading of
biomass oils to high-value chemicals and premium-grade
fuels. He reports that oxygenated compounds such as
methylphenols (cresols), methyoxyphenol (guaiacol),

Table 8 Concentrations (ppmw) of the major constituents in the ethyl acetate eluate of the flash pyrolysis oils

Reactor temperature (“C)

400 450 500 550

MW 60 (acetic acid methyl ester or glycol aldehyde) 11876 13211 12874 12000
Acetic acid propyl ester 13214 17656 14901 15710
Cyclopentanone 2231 2310 2415 2380
Cyclopentenone 4645 4650 5315 5580
Furanmethanol 2641 2460 2412 2467
Methylcyclopentenone 610 742 898 845
Furanone 7178 8141 9542 10024
Methylfuraldehyde 0 0 0 2308
Methylcyclopentenone 0 0 0 681
Phenol 937 1073 1992 1839
Methylhydroxycyclopentenone 1987 2143 2101 2470
Methylphenol 779 1213 2215 2261
Methoxyphenol 3373 3657 2892 2930
Dimethylphenol 116 149 430 436
Methylmethoxyphenol 206 177 143 1412
Benzenediol 3432 3771 3730 4028
Methylbenzenediol 715 1313 2379 2675
Trimethylphenol 0 0 408 229
Tetramethylphenol 0 0 0 0
Dimethylmethoxyphenol 1317 1021 954 750
Hydroxymethylphenylethanone 3003 2974 2854 3097
Dimethoxyphenol 3756 3720 4120 4261
Ethenylbenzenediol n/a nla nla 1414
Methoxypropenylphenol 2711 2881 3214 3542
MW 168 (unidentified) 4271 3297 3365 3252
Methoxypropenylphenol 6645 6525 4974 4511
Hydroxymethoxyphenylethanone 1316 1723 1721 1775
Dihydroxymethoxyphenylethanone 1190 1303 1021 807
Hydroxymethoxyphenylpropanone 895 1074 876 913
MW 180 (unidentified) 3134 3017 4521 5246
Dimethoxypropenylphenol 2381 2751 2552 2257
MW 180 (unidentified) 597 703 515 465
Hydroxydimethoxybenzaldehyde 1695 1723 2195 1669
Hydroxymethoxypropenylphenol 7660 8305 8778 9203
Dimethoxypropenylphenol 5834 5520 4874 4948
Hydroxydimethoxyphenylethanone 1019 1071 1066 1026
MW210 (unidentified) 641 607 688 906
MW210 (unidentified) 1444 1861 2154 3146
Naphthol 33 40 65 54
Methylnaphthol 66 70 99 116
Dimethylnaphthol 0 0 0 0
Trimethylnaphthol 0 0 0 0

Fuel 1996 Volume 75 Number 9 1057

Influence of temperature on flash pyrolysis of biomass: P. A. Horne and P. T. Williams
2-furaldehyde (furfural) and methoxypropenylphenol
(isoeugenol), which have been shown to be in high
concentration in the oils (Table 8), have a considerable
economic potential. Such chemicals have applications in
the pharmaceutical, food and paint industries.
CONCLUSIONS
The formation of pyrolytic liquid products derived
from biomass can be maximized using a fluidized bed
reactor coupled with moderate temperatures of 400-
550°C and short residence times.
The pyrolysis temperatures of 500 and 550°C gave a
pyrolytic liquid product which was homogeneous and
of low viscosity. The pyrolytic oil was found to
contain material with a molecular mass of up
to 1300 u. Chemical fractionation showed that the
pyrolytic liquids were composed almost entirely of
oxygenated components, with only low quantities
of hydrocarbons present.
The analysis of the ethyl acetate eluate from the
chemical fractionation of the pyrolytic liquids showed
that the oils contained significant quantities of
oxygenated aromatics, mainly phenol and its deriva-
tives. The concentration of phenol and its alkylated
derivatives was greatest at 500 and 550°C whereas the
concentration of the larger phenolic compounds was
greatest at lower temperatures. Some of the phenolic
compounds present have been shown to have a
significant commercial value.
PAH up to molecular mass 252 were found to be
present in the pyrolytic liquids. The concentration of
PAH was increased with temperature, but the overall
PAH concentration in all the pyrolytic liquids was
low.
The pyrolytic liquids were found to have a relatively
low CV, but they contained -63% of the potential
energy in the initial biomass feed and had a much
greater density than the original biomass.
ACKNOWLEDGEMENTS
This work was supported by the UK Science and
Engineering Research Council under grant numbers
GR/F/06074 and GR/F/87837, whose support the
authors gratefully acknowledge.
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Fuel 1996 Volume 75 Number 9 1059
 UTILIZATION OF BIOMASS PYROLYSIS FOR ENERGY PRODUCTION,
SOIL FERTILITY AND CARBON SEQUESTRATION
ROBERT HAWKINS, JON NILSSON AND REBECCA OGLESBY
(for authors correspondence, E-mail: jnilsson@c6scientific.com)

Abstract. New pyrolysis technologies have been developed that allow for carbon sequestration through
the production of sustainable energy from biomass (bioenergy). These systems produce charcoal (biochar)
and energy in the form of heat, steam, electricity, or liquid fuels. Purified hydrogen can also be produced,
allowing production of ammonia and future electric systems that utilize hydrogen (such as hydrogen fuel
cells). Pyrolysis energy systems produce more power than they consume, and can supply their own power
utilizing waste heat from the system. Therefore, this technology could be deployed without the need for
existing energy infrastructure. The biochar is a carbon-based co-product that has value as a soil
amendment, containing nutrients such as potassium (K), phosphorous (P), magnesium (Mg) and calcium
(Ca). When placed in the soil, an increase in soil organic matter (SOM) is observed, along with increases
in crop productivity, water retention, and soil biological activity as well as a decreased fertilizer
requirement. Pyrolysis technology can be deployed on a large industrial scale, or on small farm or
community scales. In these applications it can produce fuel, heat, electricity and fertilizer from crop
residues and wastes. The deployment of new biochar and bio-energy systems creates economic
opportunities for local communities through the creation of new businesses that develop to support its
infrastructure (suppliers of bio-wastes, manufacturer and distribution of co-products, and related
agricultural application services etc.). Due to its adaptability to a wide range of feedstocks, over 60
organizations are now involved in biochar research worldwide.

Biomass Transport
- manure Energy
Biofuel
- organic wastes Coproducts
bio-oil
- crop residues Industry
hydrogen
- wood waste

Pyrolysis

Residual Heat

Optionally, N2, NOX,

SOX, CO2 can be added
to increase C sink and
nutrient content

Returned to
soil as bio-char

Figure 1. Concept of low-temperature pyrolysis bio-energy with biochar sequestration.

Typically, about 50% of the pyrolyzed biomass is converted into biochar and can be returned to soil.

(Adapted from: Lehmann, J. 2007, Bioenergy in the black. Front Ecol Environ 2007; 5(7): 381–387)
Biomass Pyrolysis Technology
At the 2007 United Nations Commission on Sustainable Development, a new system of converting biomass to
energy was presented which can reduce dependence on oil. This technology is called biomass pyrolysis. In
this system, biomass is exposed to high temperatures in the absence of oxygen, producing energy and co-
products. Although pyrolysis biofuel production represents only a small portion of energy production
worldwide (UNDP 2004), it has the potential to generate electricity at a cost lower than any other biomass-to-
electricity technology available (Bridgewater et. al. 2002). A main advantage to implementing this technology
is that a pyrolysis system can supply its own power and heat by utilizing waste heat from the system, so there
is no need to supply power or heat from outside sources (Iwasaki, 2003). Therefore, these systems can be
deployed without the need for existing energy infrastructure. With these new advances, well over 15 countries
are now involved in commercializing biomass pyrolysis systems (http://terrapreta.bioenergylists.org/company).
A number of companies are working on making this technolgy more scaleable to agricultural industries with
various sizes of pyrolysis units. With these new designs, it is estimated that a 1-ton of biomass per hour unit
can produce 1 mw of electricity, 1 megawatt of usable heat and 600 pounds of charcoal per hour. A unit
capable of processing 25kg of biomass is estimated to produce 25 kw of heat and electricity and 20 pounds of
charcoal per hour.

The EPRIDA pyrolysis plant that operated at the Biomass Conversion Center in Athens, GA. until spring of 2009.

Biomass Pyrolysis vs. Conventional Biomass to Energy Systems

In conventional use of biomass for fuel, biomass is harvested and burned, and like fossil fuels, releases
compounds back to the atmosphere. This contributes to increased greenhouse gases. In order for the energy
cycle to be truly carbon neutral, an amount of biomass equal to that which was harvested must be re-grown so
that the plants can absorb an equivalent amount of CO2. To be a steady fuel supply, biomass crops require an
increase in agricultural production, which further depletes soil nutrients and minerals. This reduces the ability
to grow biomass in the future. Therefore, although biomass crops are a renewable source of energy, they are
not necessarily sustainable. With biomass pyrolysis, what was previously considered agricultural waste (crop
residues, wood wastes, manures) can create energy and nutrient enhancing soil supplements. The energy
created can be converted into several forms including hydrogen and electricity, which can be used to power
small farms or fed back onto the energy grid.

Examples of feedstocks include: coconut husks, corn stover, bean stubble, tobacco stalks, wastes from
agricultural processing, wood wastes from manufacturing and lumber industries, demolition wood wastes,
short-rotation energy crops, municipal solid waste, manure and sewage (Antal 1982). By using agricultural
wastes, biomass pyrolysis does not compete with food production.

The Products of Biomass Pyrolysis

The main products generated by biomass pyrolysis are pyrolysis vapors, heat and charcoal (biochar). These
outputs can be used in a wide range of applications.

1. Pyrolysis vapors can be condensed to form bio-oil

Bio-oil is a complex mixture of oxygenated hydrocarbons and water that can be used as low grade heating fuel.
Due to its high density, bio-oil is much more economical to transport than either biomass or hydrogen (Czernik
et al., 2007). The heating value of bio-oil is about 40% to 50% of that for petroleum-based fuels (Yaman,
2004) and about 60% of ethanol (Raveendran et al., 1996). Bio-oil can be refined to be used as a source of
chemical feedstock for gasoline, can be added to petroleum refinery feedstock or combusted in raw form
(Samolada et al., 1998). Biomass pyrolysis allows biomass to be processed at dispersed locations where wastes
are generated and bio-oil can be transported to a central refinery or power plant. Cost benefits are significant
due to the high price of transporting biomass feedstock over large distances (>30 km). Decentralized
production of bio-oil also makes sense since biomass is often generated in rural areas where bio-oil can be
processed for use in agricultural machinery.

2. Pyrolysis vapors can be used directly for energy

In this scenario it is not necessary to condense pyrolysis vapors into bio-oil to extract the energy. Pyrolysis
vapors can be burned directly as fuel for integrated heat and power production, or refined to produce fuels and
chemicals such as gasoline, diesel, alcohols, olefins, oxychemicals, synthetic natural gas and high purity
hydrogen (Magrini-Bair and others, 2007). If the energy is needed for local use, such as on a small farm, it is
better to work with the pyrolysis vapors in this form.

3. Pyrolysis vapors can be treated to produce synthetic gas (syngas)

Utilizing steam reforming, pyrolysis vapors can produce a syngas consisting of over 50% hydrogen, plus CO,
CO2, and small amounts of methane (Czernik et al., 2007). Since these gases are comprised of hydrocarbons,
they should not be emitted into the atmosphere in an unaltered state. Instead they can be converted into a clean
burning, mid BTU fuel, similar to natural gas. This can be combusted in existing engines, generators, boilers,
and turbines to produce heat, steam and electricity. Syngas is also suitable as a cooking fuel and can substitute
for propane or natural gas in uses such as home heating. High purity hydrogen from syngas can be suitable for
use in hydrogen engines, fuel cells (Czernik et al., 2007) and for production of ammonia fertilizers. The
current largest use of hydrogen in the world today is for the production of ammonia. Utilizing pyrolysis to
generate hydrogen could replace natural gas as the primary feedstock required to manufacture ammonia based
fertilizers. The production of ammonia using natural gas emits carbon dioxide into the atmosphere and fixes
the price of fertilizer to the price of natural gas. Production of ammonia from syngas could change this,
allowing the price of fertilizer to become influenced by the lower price of biomass wastes.

4. Pyrolysis syngas can create synthetic liquid fuels

Pyrolysis of biomass is one of the leading near-term options for renewable production of hydrogen and has the
potential to provide a significant fraction of transportation fuel required in the future (Czernik et al., 2007).
This can be achieved by use of hydrogen fuel cell vehicles or hydrogen powered combustion engines.
Pyrolysis syngas can be used to produce transportation fuels that work with current infrastructures and
technologies. Hydrogen and carbon monoxide, main components of the pyrolysis syngas, are the reactants
necessary to produce liquid fuels (methanol, ethanol, gasoline, aviation fuel and diesel fuel) via Fischer-
Tropsch (F-T) synthesis. F-T synthesis is regarded as the key technological component for converting syngas
to transportation fuels and other liquid products (Wilhelm and others, 2001). F-T diesel is not bio-diesel. FT
diesel is a clear liquid that gives complete combustion with no particulate emissions and has a higher energy
density that petroleum diesel and biodiesel. F-T diesel can be used in all existing diesel engines and can be
mixed without a maximum mixture level with petroleum diesel (Wilhelm and others 2001). For instance Audi
won the “24 Hours at LeMans” sports car race with F-T diesel. Currently, F-T fuels are produced from syngas
originating from natural gas and coal. Biomass syngas can replace fossil fuels as the primary feedstock.

5. Pyrolysis vapors can produce non-energy products

Pyrolysis vapors can also be used to produce a number of co-products such as wood preservative, meat
browning, food flavorings, adhesives, or specific chemical compounds (Czernik, 2004). Liquid smoke, the
chemical used to add smoke flavor to foods, is currently produced by pyrolysis of mesquite and other
hardwoods. In local agricultural applications these vapors can be condensed and used as insecticides,
herbicides, and fungicides (Steiner, 2007). The bio-oil by product from these processes can be refined as a
source of chemical feedstocks to yield products such as acetic acid (vinegar).

Figure 2. Co-products from Biomass Pyrolysis (Olglesby, Hawkins, 2007)

6. Biomass pyrolysis can create valuable soil amendments

One of the most exciting new benefits of biomass pyrolysis is its ability to produce valuable soil amendments
in the form of charcoal (biochar). Biochar is currently used in Japan and in other parts of the world by
indigenous tribes. Recent archeological exploration has found that indigenous peoples of the Amazon used
charcoal to enrich their soil over 1,000 years ago. This was due to the discovery of a black colored soil in the
Amazon basin of Brazil termed Terra Preta. It is believed that prior to the arrival of Europeans, the charcoal in
these soils was added by native Amazonians to create arable farmland (Lehmann et al., 2006). Phosphorus (P)
and calcium (Ca) are normally scarce in the very acidic Oxisols and Utisols that are predominant in this region.
In contrast, Terra Preta soils contain higher levels of P and Ca with a higher, almost neutral pH (Glaser et al.,
1998). Another distinctive feature of Terra Preta soil is the high stability of its soil organic matter (SOM), and
high cation exchange capacity (Sombroek, 2003), all factors that improve soil fertility.
 The use of charcoal as a soil amendment is not
limited to ancient civilizations such as the ones that
created Terra Preta. New research has shown that
biochar is more efficient at increasing soil fertility
and nutrient retention than un-charred organic
matter (Lehmann et al., 2006). Carbon enhanced
SOM offers direct value through improved water
infiltration, water holding capacity, structural
stability, cation exchange capacity, soil biological
activity and as a CO2 sink (Lehmann, 2007).
Charcoal can also reduce fertilizer runoff and
adsorb ammonium ions.
The use of biochar has recently been authorized for
use as a soil amendment in Japan. Of all of the
charcoal used in Japan in 1999, the highest
percentage of use was in agricultural land as a soil
Figure 3: LETS FIND A NEW PHOTO
amendment.
The second highest use was in the livestock industry where it used for animal feed and deodorization (Okimori
et al.,2003). In the U.S. a system has been developed where biochar can be amended with ammonium
bicarbonate producing a valuable carbon based fertilizer called ECOSS (Day and others, 2005).
Other benefits of biochar include its ability to: adsorb soil-damaging pesticides and neutralize natural toxins in
decomposing organic materials (Yelverton and others, 1996), and increase soil organic content (Blanco-Canqui
et al., 2004). On farm trials in the U.S., a 20% increase in corn yield and a 520% in mycorrhizal populations
(beneficial soil fungi that plants depend on) was observed where carbon based soil amendments were applied at
7-9 pounds per acre. In two years of trials at the Virginia Polytechnic Institute, a similar product achieved a 10%
increase in sweet corn yield, a 30-pound per acre savings in nitrogen for Irish potatoes and a 47% increase in
tomato yield (Morse, R and P. Stevens, 2006,-2008). Observations in the field also verified reduced need for
irrigation where carbon based amendments were applied ( http://www.carbonchar.com/). Under proper
conditions, scientists have also shown that when added to soil, biochar has the potential to increase soil carbon
sequestration by as much as 400%. This is due to its beneficial effects on soil microorganisms, which convert
soluble organic matter into stable organic compounds (Day, Reicosky, Nichols 2005).

7. Biomass pyrolysis can be used to sequester atmospheric carbon dioxide

Charcoal is commonly used for heating and cooking, and in many developing countries is the only available fuel.
In traditional methods of charcoal manufacturing all the valuable chemicals (tars, oils and smoke) and heat
escape into the atmosphere. While biomass pyrolysis can provide fuel for heating and cooking, it is vastly
different than the smoking kilns and barrels that are currently used throughout the world. Pyrolysis systems that
produce biochar and energy do not produce pollution, contaminate water supplies, or create waste disposal
problems. To ensure that systems producing biochar are clean and do not contribute to green house gas (GHG)
pollution, an organization called the International Biochar Initiative has formed and is setting standards for this
product (http://www.biochar-international.org/home.html). Biomass pyrolysis can sequester up to 50% of the
initial carbon (C) input and return it to the soil. The initial loss of C can be used for energy production and can
offset fuel use (Figure 1.). This contrasts greatly with burning of biomass, which sequesters 3% of the initial C
as charcoal, with the rest being emitted to the atmosphere, or biological decomposition which retains only 10 –
20% of initial C after 5 – 10 years (Lehmann et al., 2006). Therefore, with its ability to capture and store carbon
in the soil, biomass pyrolysis can deliver tradable carbon emission reductions (Lehmann, et.al. 2006).
Controlled pyrolysis has recently been approved by the United Nations Framework Convention on Climate
Change as a Clean Development Mechanism (CDM) for avoidance of methane production from biomass decay.
(http://cdm.unfccc.int/UserManagement/FileStorage/CDMWF_AM_C7UWTIEMRJ05M3D02XWDW80JN989IP).
In a CDM feasibility study on pyrolysis at an industrial tree plantation, it was calculated that annual processing
of 368,000 tons of biomass would provide emissions reductions of 230,000 tons of CO2 per year and could
provide jobs for approximately 2,600 people (Okimori, 2003). The latest figures published by the World Bank
indicate that the carbon market grew in value to an estimated US$30 billion in 2006 (€23 billion), three times
greater than the previous year. As of November 2007, over 850 carbon-offset projects have been registered
worldwide with about seven percent of them in the area of biomass fuels (UNEP 2008 Yearbook).
One of the first machines for offsetting CO2 emission has been in use in Senegal in the Saint-Louis region since
the end of 2007 . In partnership with Areva, the technology was transferred to South Africa to the Necsa
Company who has a production license for the southern cone of Africa. The calculation evaluating carbon credits
generated by this machine result in: 11.6 tons of CO2-equivalent per ton of green charcoal. Air France, through
the intermediary Action Carbone of GoodPlanet, is now giving its passengers the option to compensate their
CO2 emissions with carbon credits generated primarily by the Pro-Natura green charcoal project in Senegal.
(http://www.pronatura.org/index.php?lang=en&page=greenchar#greenchar2)

7. Carbon from biomass pyrolysis can be used in a wide range of non-soil applications
In addition to use for soil applications, biomass carbon can also be considered for more traditional applications.
The applications for carbon and its compounds are so widespread that it would be impossible to adequately
describe them in a single article. Some common examples include: carbon black as a pigment for printing ink,
carbon paper, printer toner, a filler in plastics and rubber, graphite as a lubricant and molding material in glass
manufacture, in electrodes for dry batteries, in electroplating, in brushes for electrical motors and as a neutron
moderator in nuclear reactors. Activated carbon is used in medicine to absorb toxins, poisons or gases from the
digestive system and in air and water purification. Due to the fact the fact that carbon can form alloys with iron,
it can also be used in steel production, in chemical reduction, case hardening, and in carbides for cutting and
grinding tools. One company that has been evaluating biochar for both soil and conventional use has found that it
can become a precursor for a wide range of carbon based products (http://c6scientific.com). Another use for
biochar that can achieve carbon emission reduction credits under the Kyoto protocol is its use as a replacement
for coal in power generation. Biochar can be used as a direct replacement for coal without the need to modify
the existing powerplant. All of the CO2 that is released as a result of replacing fossil fuels with biochar counts
toward carbon emission reductions and can be traded in the carbon exchange markets. As pyrolysis systems
become more readily available, it is important that system managers take advantage of the full range of
applications and markets that may be achieved.

Conclusions
Biomass fuels such as wood, herbaceous materials and agricultural by-products currently form the world’s third
largest primary energy resource, behind coal and oil. At best, conventional biomass to energy is considered to
be carbon neutral. Harvesting biomass to produce energy may not be sustainable because it can result in reduced
soil productivity by depletion of carbon and nutrients. Biomass pyrolysis addresses this dilemma, because it can
utilize waste products and about half of the original carbon can be returned to the soil (Lehmann, 2007).
Utilizing biomass pyrolysis for the production of fuels also has significant advantages when compared to coal
fuels because it can eliminate the need for post combustion scrubbing and can reduce nitric oxide (NOx)
formation (Bisio et al., 1995). In fact such energy is actually CARBON NEGATIVE, because for each carbon
molecule recycled back to the atmosphere, one is buried in the soil, so the net effect is to reduce atmospheric
CO2!

The deployment of biomass pyrolysis systems can create new local businesses, job opportunities and raise the
income of people in rural communities (Okimori et al., 2003). Farming communities can benefit most from this
system because the biochar co-product can reduce or eliminate purchased fertilizers while sequestering
atmospheric CO2 (Glaser and others., 2002). This can create new profit centers for landowners by creating
carbon credits and energy, which farmers can use or sell. This can decentralize fertilizer and energy distribution,
making resources more available to farmers. It can reduce agricultural dependence on petroleum and natural gas
based products by allowing regional energy production that is cost competitive with fossil fuels.

Although biomass pyrolysis represents only a small portion of energy production worldwide it has the potential
to generate energy at a lower cost than other energy systems. With its carbon negative footprint, biomass
pyrolysis has the ability to do this in a way that can contribute to reduction in greenhouse gas emissions. Given
that 1) soil organic carbon is one of the largest reservoirs in interaction with the atmosphere and 2) enhancing
natural processes is thought to be the most cost-effective means of reducing atmospheric CO2; biomass
pyrolysis provides a way forward toward overcoming the obstacles that are facing biofuels production today. In
the words of USDA Soil Scientist, David Laird, we now have “A Win–Win–Win Scenario for Simultaneously
Producing Bioenergy, Permanently Sequestering Carbon, while Improving Soil and Water Quality” (Laird,
2008).

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ORIGINAL ARTICLE
Blackwell Publishing, Ltd.

Effect of charcoal
ORIGINAL ARTICLE
on N2O emissions

Effects of charcoal addition on N2O emissions from soil resulting

from rewetting air-dried soil in short-term laboratory experiments
Yosuke YANAI1, Koki TOYOTA2 and Masanori OKAZAKI2
1
Graduate School of Bio-Applications and Systems Engineering and 2Institute of Symbiotic Science and Technology,
Tokyo University of Agriculture and Technology, Tokyo 184-8588, Japan

Abstract
Laboratory experiments were conducted to examine the effect of charcoal addition on N2O emissions
resulting from rewetting of air-dried soil. Rewetting the soil at 73% and 83% of the water-filled pore space
(WFPS) caused a N2O emission peak 6 h after the rewetting, and the cumulative N2O emissions throughout
the 120-h incubation period were 11 ± 1 and 13 ± 1 mg N m−2, respectively. However, rewetting at 64%
WFPS did not cause detectable N2O emissions (−0.016 ± 0.082 mg N m−2), suggesting a severe sensitivity to
soil moisture. When the soils were rewetted at 73% and 78% WFPS, the addition of charcoal to soil at
10 wt% supressed the N2O emissions by 89% . In contrast, the addition of the ash from the charcoal did not
suppress the N2O emissions from soil rewetted at 73% WFPS. The addition of charcoal also significantly
stimulated the N2O emissions from soil rewetted at 83% WFPS compared with the soil without charcoal
addition (P < 0.01). Moreover, the addition of KCl and K2SO4 did not show a clear difference in the N2O
emission pattern, although Cl− and SO4 , which were the major anions in the charcoal, had different
2−

effects on N2O-reducing activity. These results indicate that the suppression of N2O emissions by the
addition of charcoal may not result in stimulation of the N2O-reducing activity in the soil because of
changes in soil chemical properties.
Key words: denitrification, K fertilization, liming, N2O-reducing activity, rewetting effect.

INTRODUCTION communities and that liming and anions affected these

N2O is an important greenhouse gas produced in soil
(Bouwman 1990). It has a destructive potential in the
stratospheric ozone layer (Crutzen 1981). Nitrification
and denitrification are the most important biological
processes in the production of N2O in soil. Denitrification
is identically the sole process associated with N2O
reduction (Zumft 1997).
In a previous study, we examined the effects of soil
amendments (liming material, inorganic salts and
charcoal) on the N2O-reducing activity of denitrifying
communities (Yanai et al.). We found that charcoal and
its ash, which had a high content of alkali and inorganic
salts (Nerome et al. 2005), seemed to promote the growth
activity and N2O-reducing activity of denitrifying
Correspondence: Y. YANAI, BASE 415, 2-24-16, Nakacho,
Koganeishi, Tokyo 184-8588, Japan. Email: yosukey@
yahoo.co.jp
Received 31 July 2006.
Accepted for publication 15 December 2006.
activities more than cations. These results suggest
that N2O emissions from soil can be affected by certain
soil amendments because of the modifying activity of
N2O reduction, assuming that N2O emissions from soil
through denitrification depend on the balance of the
N2O-producing and N2O-reducing activity of denitrify-
ing communities (Cavigelli and Robertson 2001). In
fact, Inubushi et al. (1999) and Azam and Müller
(2003) observed stimulation of N2O emissions from soil
by the addition of NaCl in a laboratory incubation
study, and this result can be explained by the suppression
of N2O-reducing activity by Cl− or Na+ (Yanai et al.). In
contrast, the effects of liming on N2O emissions from
soil are inconsistent in field studies (Borken and Brumme
1997; Butterbach-Bahl et al. 1997; Klemedtsson et al.
1997; Mosier et al. 1998; Tokuda and Hayatsu 2004;
Wang et al. 1997) and in laboratory incubation studies
(Borken et al. 2000; Clough et al. 2003; Clough et al.
2004; Khalil et al. 2003).
Pulses of N2O emission have been observed in field
studies following irrigation and precipitation events

© 2007 Japanese Society of Soil Science and Plant Nutrition

182 Y. Yanai et al.

(e.g. Kusa et al. 2006; Ruser et al. 2001) and have been
reproduced in laboratory incubation experiments as a
rewetting of dry soil (Rudaz et al. 1991). Rudaz et al.
(1991) and Ruser et al. (2006) investigated the contri-
bution of nitrification and denitrification to the produc-
tion of N2O emitted after rewetting using the C2H2
addition method and 15N tracing technique, respectively,
and concluded that the N2O was mainly produced
through denitrification. In the present study, to examine
the relationship between enhancing the N2O-reducing
activity of denitrifying communities and N2O emissions
from soil, we examined the effects of charcoal and
anion species on N2O emissions caused by the rewetting
of air-dried soil in the laboratory.
MATERIALS AND METHODS
Soil samples and charcoal
The soil samples examined were the same as those used
in our previous study (Yanai et al.). Soil sampling was
conducted at the Field Museum Tsukui, the Field
Science Center of Tokyo University of Agriculture and
Technology, Tokyo, Japan. Soil samples were collected
from a grassland field in which one side was planted
with Sorghum bicolor (L.) Moench and the other with
Sorghum sudanense (Piper) on April 2004 and March
2005, respectively. The soil is classified as Typic
Hapludand, and the soil texture is loam to clay loam at
a depth of 0–40 cm, with a granular structure (Kurokawa,
pers. comm.). After collection, moist soil samples were
passed through a 2-mm mesh sieve, and part of the
sample was then air-dried. Selected physico-chemical
properties of the soil samples are listed in Table 1. Soil
pH (H2O) value was determined in a 1:2.5 air-dried soil
(weight) to deionized water (volume) ratio. Total carbon
and nitrogen contents of the soils were determined
using the dry combustion method using a CN CORDER
MT-700 (Yanaco, Kyoto, Japan). Water soluble organic C
−
and NO3-N contents were determined using a TOC
meter (TOC-VCSH, SHIMADZU Co. Ltd., Kyoto, Japan)
and an ion chromatograph (LC-20AT, SHIMADZU Co.
Ltd., Kyoto, Japan), respectively, in 1:10 extracts (air-dried
soil to deionized water w/v) at 240 rpm for 30 min. The
population density of denitrifiers in the air-dried soil
samples was determined using the most probable number
method in five replicates of 10-fold serial dilution (Tiedje
1994). Maximum water-holding capacity (MWHC) was
determined using the Hilgard method. Particle density
was determined using the pycnometer method (Blake
and Hartge 1986). Water-filled pore space (WFPS) was
calculated as follows:
WFPS = (Gravimetric water content/ρH2O) ·
(Bulk density/Porosity)
where Porosity = 1 − (Bulk density/Particle density). In the
present study, we set the density of water (ρH2O) at 1 g cm−3.
The charcoal, which was made from municipal biowaste,
was provided by JFE Holdings. The physico-chemical
properties of the charcoal are listed in Yanai et al. and
its potential usefulness for cultivation was demonstrated
by Nerome et al. (2005). Some selected physico-chemical
properties of the charcoal and its ash, which was obtained
by heating at 700°C for 4 h (as the test of weight loss-on-
ignition; LOI), are listed in Table 2. Charcoal and ash
pH (H2O) values were determined in a 1:5 air-dried
material to deionized water ratio (w/v). The MWHC and
particle density were determined as described above. Anion

Table 1 Selected physico-chemical properties of the air-dried soils examined in this study (oven-dry basis)

Water Bulk Specific

−
Total C Total N WSOC NO3 -N Denitrifiers content MWHC density gravity Porosity
Abbreviation Date of pH C/N
of soil name sampling (H2O) (mg C g−1) (mg N g−1) ratio (µg g )−1
(log MPN g )−1 −1
(g H2O g ) −3
(g cm ) (cm3 cm−3)

TG2004 Apr. 2004 6.0 69.6 5.62 12.4 155 45.8 6.5 0.15 1.21 0.58 2.05 0.72
TG2005 Mar. 2005 5.4 70.3 5.45 12.9 74 6.4 6.2 0.13 1.11 0.59 2.03 0.71

MPN, most probable number; MWHC, maximum water-holding capacity; WSOC, water soluble organic carbon.

Table 2 Selected physico-chemical properties of charcoal and its ash examined in this study

Anion content
Water Bulk Particle
(µmol g−1)‡
pH LOI content MWHC density density
−
(H2O)† (%) (g H2O g−1) (g H2O g−1) (g cm−3) (g cm−3) Cl− NO3 SO4
2−

Charcoal 9.3 38 0.14 1.38 0.50 1.64 510 0 9

Ash 11.6 – 0.03 – 0.46 – 1240 2 80
†
1:5 ratio. ‡Air-dried material basis in Yanai et al. LOI, weight loss-on-ignition; MWHC, maximum water-holding capacity; –, not determined.

© 2007 Japanese Society of Soil Science and Plant Nutrition


−
contents (Cl−, NO3 and SO4 ) in these materials were
2−
determined using an ion chromatograph (LC-20AT,
SHIMADZU Co. Ltd. Kyoto, Japan) in 1:20 extracts
(air-dried material to deionized water w/v) at 240 rpm
for 30 min.
Measurement of N2O emissions from soil
after rewetting
To simulate the thin surface layer in arable fields, where
the soil could be subjected to air-drying following
continuous clear weather, 30 g of sieved air-dried soil
was placed in a Petri dish (1.3 cm height and 8.5 cm
diameter) without compaction. As a result, the thickness
of the soil was approximately 0.8 cm. To simulate the
condition of the soil during or immediately after precip-
itation, distilled water was added into the soil samples
in the Petri dishes to more than 70% of their MWHC
(equivalent to 64% of the water-filled pore space [WFPS]).
Immediately after rewetting the soil sample, N2O
emissions were periodically measured using the closed-
chamber method (Hutchinson and Mosier 1981). A
clear glass bell-jar (14 cm width, 26 cm height, 2.32 L)
was used as a gas-tight chamber to monitor the concen-
tration change in the headspace gas. The inlet and outlet
of the bell-jar were sealed with a rubber stopper and a
rubber septum, respectively, and the bottom part of the
bell-jar was tightly attached with a ground glass-plate
using a high vacuum-sealing compound (HIVAC-G,
Shin-Etsu Chemical Co. Ltd., Tokyo, Japan). A pressure-
controlling bent (Hutchinson and Mosier 1981) was
installed at the rubber stopper in the inlet and gas samples
were collected through the rubber septum installed at the
outlet. After placing the chamber onto the soil sample,
headspace gas was withdrawn five times at 2 or 8 min
intervals, depending on the rate of concentration change.
N2O concentration in the collected gas sample was analyzed
using a gas chromatograph (GC-14A, SHIMADZU,
Kyoto, Japan) equipped with an electron capture detector
and a stainless steel column packed with Porapak-Q (80/
100 mesh, 3 mm diameter, 2 m length). The column and
detector temperatures were kept at 90°C and 330°C,
respectively. Argon containing 5% CH4 was used as a
carrier gas at a flow rate of 23 mL min−1. The N2O emis-
sion rate was calculated using the linear regression method
(Hutchinson and Mosier 1981). After measurement of
the N2O emission rate, the chamber was removed and the
soil sample was left at room temperature (approximately
20–28°C) without a lid on the Petri dish. The water
content was maintained during the incubation period
by adding distilled water. N2O emissions were monitored
until the first peak of N2O emissions disappeared.
As a preliminary experiment, we examined the effect
of moisture content after rewetting on the N2O emissions
from soil (TG2005). Distilled water was added into the
© 2007 Japanese Society of Soil Science and Plant Nutrition
Effect of charcoal on N2O emissions 183
soil samples at 17.0, 19.9 and 23.1 mL to adjust the
rewetted condition to 70, 80 and 91% MWHC, equivalent
to 64, 73 and 83% WFPS, respectively. This experiment
was conducted in triplicate.
Effect of soil amendments on N2O emissions
from soil resulting from rewetting
Based on the result of the preliminary experiment (Fig. 1),
the moisture content after rewetting was adjusted to
more than 73% WFPS in this study. First, to simulate
precipitation in grassland amended with charcoal in the
surface layer, 2 mm-sieved charcoal was mixed with
soil (TG2004) in three of six Petri dishes at 10 wt%
(equivalent to 13 vol%) before rewetting, and the N2O
emissions were compared with the remaining three Petri
dishes as the non-added control. The soil samples were
added with distilled water to moisten the soil of the
non-added control at 78% WFPS. Second, to test whether
the effect of the charcoal addition on the N2O emission
results from the stimulation of N2O-reducing activity by
pH increase, the charcoal or its ash was mixed with soil
(TG2005) in three of nine Petri dishes before rewetting,
and three Petri dishes as the non-added control. The
amounts of added charcoal and its ash were determined
in order to set soil pH (H2O) at 6.0, and the rate of
addition was 8.2 and 1.6 wt%, equivalent to 9.7 and
2.0 vol%, respectively. The soil samples were rewetted
by adding distilled water, which was necessary to
Figure 1 Effect of rewetting on N2O emissions from soil
(TG2005). An air-dried soil sample was rewetted using
distilled water at 64 (×), 73 (
) and 83% () of the water-filled
pore space (WFPS) and incubated at room temperature. The
values shown are the mean ± standard deviation of three replicates.
The cumulative N2O emissions during the 120-h incubation
period at a rewetting level of 64, 73 and 83% WFPS were
−0.016 ± 0.082, 11 ± 1 and 13 ± 1 mg N m−2 (equivalent to
−0.003 ± 0.03, 2.3 ± 0.3 and 2.8 ± 0.4 µg N g−1soil), respectively.
184 Y. Yanai et al.

moisten the soil of the non-added control at 73% WFPS.
Third, to estimate the interaction between the rate of
charcoal addition and the moisture content after rewet-
ting, N2O emissions were compared with three levels of
charcoal additions (0, 2 and 8.2 wt%, equivalent to 0,
2.4 and 9.7 vol%, respectively) in triplicate. Distilled
water was added to the soil samples to moisten the soil
of the non-added control (0% charcoal) at 83% WFPS.
Finally, as Cl− and SO2− 4
were not only the major anion
species of the charcoal (Table 2), but also were applied
into arable fields through fertilization, we examined the
effect of anion species of K solution on N2O emissions
after rewetting. Of the nine Petri dishes containing the
soil samples (TG2005), distilled water, 10 mmol L−1 KCl
and 5 mmol L−1 K2SO4 solution were each added to three
dishes to adjust to 73% WFPS of the soil, and the N2O
emissions were compared. The concentration of K solution
was decided based on the concentration of K in a commer-
cial liquid fertilizer (Otsuka Chemical Co. Ltd., Osaka,
Japan), and the estimated load of Cl− and SO4 added with
2−
83% WFPS, and the cumulative N2O emissions throughout
the 120-h incubation period at room temperature
were −0.016 ± 0.082, 11 ± 1 and 13 ± 1 mg N2O-N m−2
(−0.003 ± 0.03, 2.3 ± 0.3 and 2.8 ± 0.4 µg N2O-N g−1soil),
respectively (Fig. 1). Rewetting over 73% WFPS triggered
N2O emissions, but there were no significant differences
in the cumulative N2O emissions between soils rewetted
at 73% and 83% WFPS (P = 0.180).
Effects of charcoal addition on N2O emissions
after rewetting at 73% WFPS for TG2004
The highest N2O emission rate was observed 30 h
after rewetting, and the values were 2620 ± 460 and
383 ± 74 µg N m−2 h−1 in the treatments without and with
charcoal addition, respectively (Fig. 2). The addition of

the charcoal, ash and K solution is listed in Table 3.

Calculation of the cumulative N2O emission

and statistical analysis
The cumulative N2O emissions were estimated using the
linear trapezoidal method, and the value was expressed
as an arithmetic mean and standard deviation (SD). The
level of significance of the treatments was examined
using an unpaired t-test for TG2004 and by anova
followed by Tukey’s multiple comparison tests for
TG2005 (P < 0.05). If one of the mean values of the tri-
plicates appeared to lose normality (mean − 2SD < 0),
the original data were log-transformed before compari-
son (Bland and Peacock 2002).
Figure 2 Effect of charcoal addition on N2O emissions from
soil (TG2004) rewetted at 78% of the water-filled pore space
RESULTS of the soil. The values shown are the mean ± standard
deviation of three replicates. The cumulative N 2O emis-
N2O emissions from soil after rewetting sions during the 168-h incubation period for the non-added
control and the 10 wt% charcoal addition were 105 ± 14
N2O emissions were not detected after rewetting at 64% and 11.1 ± 2.4 mg N m−2 (equivalent to 19.9 ± 2.7 and
WFPS, but were detected after rewetting at 73% and 2.1 ± 0.5 µg N g−1 soil), respectively.

Table 3 Estimation of Cl− and SO4 load onto soil (TG2005) by the addition of charcoal, ash and K solution
2−

Concentration in soil solution (mmol L−1) at

Added into soil
(µmol g−1 soil) 73% WFPS 83% WFPS
Application rate or
– 2− − 2− − 2−
concentration Cl SO 4 Cl SO 4 Cl SO4
Charcoal 2 wt% 10 0.2 NA NA 10 0.2
Charcoal 8.2 wt% 42 0.7 46 0.8 40 0.7
Ash 1.6 wt% 20 1.3 23 1.5 NA NA
KCl 10 mmol L−1 7.5 0 8.5 0 NA NA
K2SO4 5 mmol L−1 0 3.7 0 4.3 NA NA

NA, not applicable with respect to the objectives of this study; WFPS, water-filled pore space.

© 2007 Japanese Society of Soil Science and Plant Nutrition


charcoal decreased the N2O emission peak by 85% of
that of the control without charcoal. The cumulative
N2O emissions were 105 ± 14 and 11.1 ± 2.4 mg N m−2
(19.9 ± 2.7 and 2.1 ± 0.5 µg N g−1 soil) in the treatments
without and with charcoal addition, respectively. The
charcoal addition significantly decreased N2O emissions
by 89% of the control value without charcoal (P < 0.01).
Effects of liming (pH 6.0) with charcoal and its
ash on N2O emissions after rewetting at 73%
WFPS for TG2005
The highest N2O emission rate was observed at 12 h
after rewetting in the non-added control and the ash-
added soil (Fig. 3), but N2O emissions were kept at a
low level in the charcoal-added soil throughout the
observation period (72 h). The cumulative N2O
emissions throughout the 72-h incubation period in the
non-added control, ash-added and charcoal-added soils
were 4.1 ± 1.9, 4.3 ± 1.2 and 0.8 ± 0.7 mg N m−2 (0.9 ± 0.4,
1.0 ± 0.3 and 0.2 ± 0.2 µg N g−1 soil), respectively.
Charcoal addition decreased N2O emissions by 80% of
the value of the non-added control (P < 0.05), whereas
ash addition did not.
Effects of charcoal addition on N2O emissions
after rewetting at 83% WFPS for TG2005
The N2O emission rate at 6 h after rewetting was lower
in the 2 and 8.2 wt% charcoal added-soils than in the
Figure 3 Effect of liming by using charcoal and its ash on N 2O
emissions from soil (TG2005) rewetted at 73% of its water-
filled pore space. The values shown are the mean ± standard
deviation of three replicates. The cumulative N 2O emissions
during the 72-h incubation period for the non-added control,
ash-amended soil and charcoal-amended soil were 4.1 ± 1.9,
4.3 ± 1.2 and 0.8 ± 0.7 mg N m−2 (equivalent to 0.9 ± 0.4,
1.0 ± 0.3 and 0.2 ± 0.2 µg N g−1soil), respectively.
© 2007 Japanese Society of Soil Science and Plant Nutrition
Effect of charcoal on N2O emissions 185
Figure 4 Effect of charcoal addition on N2O emissions from
soil (TG2005) rewetted at 83% of its water-filled pore space.
The values shown are the mean ± standard deviation of
three replicates. The cumulative N 2O emissions during the
72-h incubation period for the non-added control and the 2
and 8.2 wt% charcoal additions were 6.8 ± 0.9, 10.0 ± 0.8
and 10.3 ± 0.6 mg N m−2 (equivalent to 1.5 ± 0.2, 2.2 ± 0.2 and
2.4 ± 0.1 µg N g−1soil), respectively.
non-added control, while the N2O emission rate more
than 12 h after the rewetting was higher in the 2 and
8.2 wt% charcoal added-soil than in the non-added
control (Fig. 4). The cumulative N2O emissions throughout
the 72-h incubation period in the non-added control,
and in the 2 and 8.2 wt% charcoal-added soil were
6.8 ± 0.9, 10.0 ± 0.8 and 10.3 ± 0.6 mg N m−2 (1.5 ± 0.2,
2.2 ± 0.2 and 2.4 ± 0.1 µg N g−1soil), respectively. The
addition of charcoal at 2 and 8.2 wt% significantly
increased N2O emissions by 47% and 51% of the values
of the non-added control, respectively (P < 0.01).
Effects of KCl and K2SO4 on N2O emissions
after rewetting at 73% WFPS for TG2005
The highest N2O emission rate was observed 12 h after
rewetting (Fig. 5). The mean N2O emission rate was
higher in 10 mmol L−1 KCl than in 5 mmol L−1 K2SO4
and the non-added control, but considerable variability
was observed in the KCl-added soil. The cumulative
N2O emissions throughout the 72 h incubation in the
non-added control, 10 mmol L−1 KCl and 5 mmol L−1
K2SO4 were 2.9 ± 0.6, 5.3 ± 6.1 and 4.4 ± 1.5 mg N m−2
(0.6 ± 0.1, 1.2 ± 1.3, 1.0 ± 0.3 µg N g−1 soil) with CV
values of 21, 115 and 34% (17, 108 and 30%), respectively.
There were no significant differences between the
control and the 10 mmol L−1 KCl (P = 0.9986) or 5 mmol L−1
K2SO4 additions (P = 0.8559), or between the 10 mmol L−1
KCl and 5 mmol L−1 K2SO4 additions (P = 0.8794).
186 Y. Yanai et al.
Figure 5 Effect of Cl− and SO4 of K salts on N2O emissions
2−
from soil (TG2005) rewetted at 73% of its water-filled pore
space. An air-dried soil sample was rewetted using distilled
water (control) or a K solution and incubated at room
temperature. The values shown are the mean ± standard
deviation of three replicates. The cumulative N 2O emissions
during the 72-h incubation period for the non-added control,
the 10 mmol L−1 KCl-added soil and the 5 mmol L−1 K2SO4
added soil were 2.9 ± 0.6, 5.3 ± 6.1 and 4.4 ± 1.5 mg N m−2
(equivalent to 0.6 ± 0.1, 1.2 ± 1.3 and 1.0 ± 0.3 µg N g−1 soil),
respectively.
DISCUSSION
The present study demonstrated that rewetting of
air-dried soil at 73% WFPS caused significant N2O
emissions (Fig. 1), and the N2O emissions were sup-
pressed by the addition of charcoal (Figs 2,3). This
suppression of the N2O emissions was first considered
to be a liming effect because charcoal has alkali (Table 2)
and it had the potential to increase the N2O-reducing
activity of denitrifying communities (Cavigelli and
Robertson 2000), which might cause a decrease in N2O
emissions (Cavigelli and Robertson 2001). Therefore,
liming resulting from the ash was expected to have a
similar potential for promoting N2O-reducing activity
to the charcoal itself. We checked the soil pH (1:2.5
ratio) after the observation of N2O emission from soils
to which charcoal and its ash had been added and there
were no significant differences between these amendments
(5.7 ± 0.03 and 5.6 ± 0.01, respectively, P = 0.08), but
these treatments were significantly different from the
non-added control (4.9 ± 0.2, P < 0.01, n = 3). However,
the addition of ash did not suppress N2O emissions (Fig. 4).
Moreover, the suppressive effects of charcoal addition
on N2O emissions were not observed when the soils
were rewetted at 83% WFPS (Fig. 4). These results
indicate that soil pH amendments, which are intended
to stimulate the N2O-reducing activity, may not explain
the suppression of the N2O emissions from soil rewet-
ted at 73% WFPS (Figs 2,3). In addition, irrespective of
the inhibitory effects of Cl− and the stimulatory effects
2−
of SO4 on N2O-reducing activity of denitrifying com-
munities (Yanai et al.), there were no clear differences in
the N2O emissions when KCl and K2SO4 were added
to the soils (Fig. 5). This finding could result from the
use of concentrations (5 and 10 mmol L−1) that were
too low to affect the denitrifying communities (Table 3)
because the effects of Cl− and were detected at more
than 40 mmol L−1 in the liquid medium in our previous
study (Yanai et al.). Nevertheless, these results sug-
gested that amelioration of the chemical properties of
soil in order to stimulate the N2O-reducing activity may
not be related to the suppression of the N2O emissions
from soil rewetted at 73% WFPS (Figs 2,3).
Increases in N2O emission rates with increasing soil
water contents have been reported from laboratory and
field studies and have been attributed to increasing
denitrifying activity induced by decreased O2 diffusion
into the soil (Ruser et al. 2006 and references therein).
In the present study, we observed a similar trend, namely,
that N2O emissions increased with increases in the water
content of soil by rewetting at 73% and 83% WFPS,
whereas significant N2O emissions were not detected by
rewetting at 64% WFPS (Fig. 1). This result suggests that
a decrease in the moisture conditions from 83% to 73%
WFPS did not affect the denitrifying communities, while
a decrease from 73% to 64% WFPS may result in a sig-
nificant decrease in the anoxic microsites, which results
in the suppression of denitrification. Thus, undetectable
N2O emissions from soil rewetted at 64% WFPS may not
be the result of complete denitrification, including N2O
reduction to N2, but, rather, to insufficient development
of anoxic microsites in the soil to trigger denitrification.
Possibly, this was caused by the soil sample TG2005,
which had less denitrification activity because of a lower
population density of denitrifiers, soil pH, and the amount
of substrate compared with the soil sample TG2004
(Table 1). In addition, the decay of the N2O emission rate
in the later incubation period after rewetting may be the
completion of N2O production (stepwise reductions of
− −
NO3, NO2, and NO) rather than the kinetic equilibra-
tion of N2O production and reduction followed by N2
production. Ruser et al. (2006) observed few N2 emissions
after rewetting, indicating a low or undetectable contri-
bution of N2O-reducing activity in the later incubation
period after rewetting. Therefore, the N2O-reducing activity
of denitrifying communities may not significantly affect
N2O emissions after rewetting of air-dried soil, suggesting
that the suppressive effect of the charcoal addition on
N2O emissions (Figs 2,3) might result from inhibition of
N2O-producing activity of denitrifying communities.
© 2007 Japanese Society of Soil Science and Plant Nutrition

Although there was no direct evidence to show a
linkage between the addition of charcoal and the
suppression of N2O emissions from soil (Figs 2,3), the
added charcoal itself probably absorbed water and
improved the aeration of the soil, leading to a suppression
− −
of N2O production (stepwise reduction of NO3 , NO2
and NO) similar to the soil rewetted at 64% WFPS (Fig. 1).
In fact, the charcoal examined was made up of porous
particles, whereas the ash was nearly pulverized. Such
differences in the size and structure possibly affect the water
absorption capacity of these materials, and may conse-
quently cause differences in the soil aeration, the denitrifi-
cation process, and N2O emissions from soil, although
the charcoal addition did not significantly affect the
MWHC or the particle density (data not shown). Hence,
the significant increases in N2O emissions by the addi-
tion of charcoal to soil rewetted at 83% WFPS (Fig. 4)
can be interpreted as an interaction between the insig-
nificant improvement of the aeration of the soil and the
stimulation of the N2O-producing activity resulting
from neutralization (e.g. Cavigelli and Robertson 2000).
Charcoal was examined in this study because of its
potential use for soil amendments in temperate regions
(Nerome et al. 2005) and in the tropics (Glaser et al. 2002;
Yamato et al. 2006). Although any extrapolation of the
findings from this short-term laboratory study to a long-
term field scale should be conducted with caution, field
applications of charcoal possibly suppress N2O emissions
from arable soil, depending on the moisture or aeration
conditions of the soil. In contrast, our understanding of the
process of suppressing N2O emissions from soil by charcoal is
still preliminary. Therefore, further studies are necessary to
understand both the mechanisms and possible side-effects
of charcoal addition to soil on the suppression of N2O
−
emissions from soil, such as the activity of NO3 assimila-
− Delwiche, pp. 17– 44. John Wiley, New York.
tion, NO2 accumulation in soil or NOx emissions from soil.
ACKNOWLEDGMENTS
The authors thank Dr Yuzo Kurokawa (Tokyo University
of Agriculture and Technology) for providing soil samples
and Mr Sumio Yamada (JFE Holdings) for providing the
charcoal samples. The work described in this report was
financially supported by a Sasakawa Scientific Research
Grant from The Japan Science Society (16-315), the TUA&T
21 Century COE program (Evolution and Survival of
Technology-based Civilization: Professor Masayuki Horio)
and by the Japan Society for the Promotion of Science
Research Fellowships for Young Scientists (17-6518).
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© 2007 Japanese Society of Soil Science and Plant Nutrition
 US Offset Market
•  Trends
– Fossil fuels will likely be capped
– Favor domestic projects over international projects

•  Destroying methane emissions

•  Sequestration
–  Forestry
–  Soil Management
–  Biochar?
 Sequestration Projects
•  Clean Development Mechanism
–  Reforestation/afforestation only
–  Temporary credits
Expected CERs Until 2012 (%) in each category

Afforestation &
Demand-side EE Reforestation
1% 0.4%

Fuel switch Transport

7% 0.2%

Supply-side EE HFCs, PFCs & N2O

10% reduction
27%

CH4 reduction & Cement &

Coal mine/bed
20%
Renewables
35%
 Outline

•  Theory of offsets
•  How offsets could support biochar projects
•  Methodology issues
•  Current funding from The Climate Trust
$ = CO2 Sequestered x Price
• Low Quality US Projects
•  Chicago Climate Exchange: $2-$4/mt CO2
•  High Quality US Projects
•  Voluntary Carbon Standard: $4-$9/mt CO2
•  California Climate Action Registry: $5-$11/mt CO2
•  Mature International Markets
•  EU Emissions Trading Scheme: $10-$40/mt CO2
•  Projections for Early US Market
•  Markey-Waxman Bill : $10-$15/mt CO2
Biochar GHG Reductions
GHG Reduction Reduction Size Qualify for Carbon
Finance?
Sequestration Large Yes

Fuel switch Medium Likely to be capped

Less fertilizer  fewer Medium/Large Hard to measure

soil emissions
(N2O and CH4)
Less fertilizer  less Small/Medium Indirect, hard to
fertilizer production measure, likely to be
capped
 Currently Methodologies

Avoidance of methane production from

biomass decay through controlled
pyrolysis

•  Small scale
•  No credit for carbon sequestration, but…
•  Char must be “biologically inert”
•  Volatile C/Fixed C ratio lower than 50%
Next Step: Sequestered Carbon

Carbon Gold methodology for proposed to

the Voluntary Carbon Standard
 Outline

•  Theory of offsets
•  How offsets could support biochar projects
•  Methodology issues
•  Current funding from The Climate Trust
 Unresolved Methodology
Issues
•  Recalcitrance
•  Guarantee 100 years of permanent
sequestration
•  Carbon Gold: Volatile C/Fixed C ratio
lower than 50%
•  Soil monitoring
•  What happens to char that erodes out of
the soils?
 Unresolved Methodology
Issues
•  Ownership – Three entities, same reduction
1.  Feedstock owner
2.  Pyrolysis plant
3.  Land owner

•  Carbon Gold: Credits pyrolysis plant

•  Sequestered carbon can only be claimed once
 Unresolved Methodology
Issues
•  Environmental impact
–  Heavy metals
–  Criteria air pollutants
–  Microbe health
–  Carbon already in soil
 Resolved Issue: Waste
Feedstocks
•  Leakage
–  Changes in emissions outside the project
itself
•  Direct: Biomass fuel unavailable
•  Indirect: displace current farm land for biochar
feedstock plantations  land use change
•  Carbon Gold: “biomass that would otherwise
have been left to decay or been burned in an
uncontrolled manner”
Carbon Markets and
Biochar: An Offset
Buyers Perspective
North American Biochar Conference
August 10th, 2009

Peter Weisberg
Offset Project Analyst
pweisberg@climatetrust.org
503-238-1915
 Outline

•  Theory of offsets
•  How offsets could support biochar projects
•  Methodology issues
•  Current funding from The Climate Trust
Source: The McKinsey Quarterly. 2007. “A cost curve for greenhouse gas reductions.”
 US Offset Market
•  Trends
– Fossil fuels will likely be capped
– Favor domestic projects over international projects

•  Destroying methane emissions

•  Sequestration
–  Forestry
–  Soil Management
–  Biochar?
 Sequestration Projects
•  Clean Development Mechanism
–  Reforestation/afforestation only
–  Temporary credits
Expected CERs Until 2012 (%) in each category

Afforestation &
Demand-side EE Reforestation
1% 0.4%

Fuel switch Transport

7% 0.2%

Supply-side EE HFCs, PFCs & N2O

10% reduction
27%

CH4 reduction & Cement &

Coal mine/bed
20%
Renewables
35%
 Outline

•  Theory of offsets
•  How offsets could support biochar projects
•  Methodology issues
•  Current funding from The Climate Trust
$ = CO2 Sequestered x Price
• Low Quality US Projects
•  Chicago Climate Exchange: $2-$4/mt CO2
•  High Quality US Projects
•  Voluntary Carbon Standard: $4-$9/mt CO2
•  California Climate Action Registry: $5-$11/mt CO2
•  Mature International Markets
•  EU Emissions Trading Scheme: $10-$40/mt CO2
•  Projections for Early US Market
•  Markey-Waxman Bill : $10-$15/mt CO2
Biochar GHG Reductions
GHG Reduction Reduction Size Qualify for Carbon
Finance?
Sequestration Large Yes

Fuel switch Medium Likely to be capped

Less fertilizer  fewer Medium/Large Hard to measure

soil emissions
(N2O and CH4)
Less fertilizer  less Small/Medium Indirect, hard to
fertilizer production measure, likely to be
capped
 Currently Methodologies

Avoidance of methane production from

biomass decay through controlled
pyrolysis

•  Small scale
•  No credit for carbon sequestration, but…
•  Char must be “biologically inert”
•  Volatile C/Fixed C ratio lower than 50%
Next Step: Sequestered Carbon

Carbon Gold methodology for proposed to

the Voluntary Carbon Standard
 Outline

•  Theory of offsets
•  How offsets could support biochar projects
•  Methodology issues
•  Current funding from The Climate Trust
 Unresolved Methodology
Issues
•  Recalcitrance
•  Guarantee 100 years of permanent
sequestration
•  Carbon Gold: Volatile C/Fixed C ratio
lower than 50%
•  Soil monitoring
•  What happens to char that erodes out of
the soils?
 Unresolved Methodology
Issues
•  Ownership – Three entities, same reduction
1.  Feedstock owner
2.  Pyrolysis plant
3.  Land owner

•  Carbon Gold: Credits pyrolysis plant

•  Sequestered carbon can only be claimed once
 Unresolved Methodology
Issues
•  Environmental impact
–  Heavy metals
–  Criteria air pollutants
–  Microbe health
–  Carbon already in soil
 Resolved Issue: Waste
Feedstocks
•  Leakage
–  Changes in emissions outside the project
itself
•  Direct: Biomass fuel unavailable
•  Indirect: displace current farm land for biochar
feedstock plantations  land use change
•  Carbon Gold: “biomass that would otherwise
have been left to decay or been burned in an
uncontrolled manner”
 Outline

•  Theory of offsets
•  How offsets could support biochar projects
•  Methodology issues
•  Current funding from The Climate Trust
•  3 Programs
•  Oregon Program
•  Smart Energy
•  Colorado Carbon Fund

•  16 projects, $8.8 million in

funding, 2.6 million tons of CO2
offset
•  Non-profit
- Laboratory for innovative
offset projects
 Project Development Timeline

Upfront Payment

Annual
Contract
Proposal Due Commercial Monitoring
Negotiation
Diligence Operation

Annual payment
“upon delivery”
Peter Weisberg
Offset Project Analyst
pweisberg@climatetrust.org
503-238-1915 x 207
 Chemosphere 67 (2007) 1033–1042
www.elsevier.com/locate/chemosphere

Differential sorption behaviour of aromatic hydrocarbons on

charcoals prepared at different temperatures from grass and wood
a,b,*,1
Ludger C. Bornemann , Rai S. Kookana a, Gerhard Welp b

a
CSIRO Land and Water, Adelaide Laboratory, Waite Road, Urrbrae SA, Australia
b
Institute of Crop Science and Resource Conservation of the University of Bonn, Division of Soil Science, Nussallee 13, 53115 Bonn, Germany

Received 27 June 2006; received in revised form 13 October 2006; accepted 19 October 2006
Available online 8 December 2006

In memory of the 100th anniversary of the birth of Prof. Dr. H.C. Eduard Mückenhausen (February 17th 1907)

Abstract

Naturally occurring charcoals are increasingly being recognized as effective sorbents for organic compounds. In this study we inves-
tigated the sorption of benzene and toluene in single-sorbate and bi-sorbate systems on different types of charcoals produced in labora-
tory, employing the batch sorption technique. Air dried plant materials from Phalaris grass (Phalaris tuberosa) and Red Gum wood
(Eucalyptus camadulensis) were combusted under limited oxygen supply at 250 C, 450 C, and 850 C. The resulting charcoals were char-
acterized for their specific surface areas, total cation content, and pore size distributions (pore size distribution only for wood combusted
at 450 C and 850 C). For the materials treated at 850 C not only the surface area, microporosity, and total amount of sorbed sorbate
increased markedly, but also the non-linearity of the sorption isotherm. The pore size distributions and surface areas as well as an indif-
ferent sorption behaviour and competition effects for both sorbates indicated that pore filling mechanisms were the dominating processes
governing the sorption on these microporous, high temperature treated materials. For the materials treated at lower temperatures the
affinity of toluene was higher compared to that of benzene. In the bi-sorbate system the overall uptake of benzene increased. These phe-
nomena might be due to the stronger hydrophobicity of toluene, and to a varying potential for swelling of the matrix and pore defor-
mation by the two sorbates. The significantly lower sorption capacity of the Phalaris-derived material compared to the Red Gum
charcoal correlated with its smaller surface area and higher cation content.
 2006 Elsevier Ltd. All rights reserved.

Keywords: Charcoal; Aromatic hydrocarbons; Pore filling; Hydrophobic partitioning

1. Introduction ment, and oxidation of charcoal, but there are indications

Charcoal, the solid residue of partially combusted bio-
mass, is a major constituent of the non-living organic mat-
ter of many soils and sediments (Schmidt and Noack,
2000). Together with other species of pyrolytic carbon it
comprises a ubiquitous group of materials in our environ-
ment, commonly named black carbon (BC) (Goldberg,
1985). Currently, little is known about formation, move-
*
Corresponding author. Tel.: +49 228 73 9368; fax: +49 228 73 2782.
E-mail address: Ludger.Bornemann@uni-bonn.de (L.C. Bornemann).
1
PMB 2, Glen Osmond, SA 5064, Australia.
that frequent vegetation fires and alluvial transport are
playing a major role (Skjemstad et al., 1997). Charcoal
withstands biological and chemical degradation to a con-
siderable degree (Goldberg, 1985) and plays an important
role in the global carbon cycle and carbon sequestration
due to accumulation processes (Jenkinson, 1990).
Several studies revealed the potential of charcoal to
serve as a strong sorbent for environmental pollutants,
thereby playing a crucial role in governing their environ-
mental fate and risks to human and ecosystem health. Poly-
cyclic aromatic hydrocarbons (PAH), polychlorinated
biphenyls (PCB), and polychlorinated dibenzo-dioxins

0045-6535/$ - see front matter  2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.chemosphere.2006.10.052
1034 L.C. Bornemann et al. / Chemosphere 67 (2007) 1033–1042
(PCDD) have been detected on carbonaceous geosorbents
and are contaminants of environmental concern (e.g.
Accardi-Dey and Gschwend, 2003; Abelmann et al.,
2005; Lohmann et al., 2005). However, the majority of
studies dealing with sorption on activated carbon, soot,
coal, charcoal, graphite, opaque particles and other forms
of BC has been conducted utilizing rather simple aromatic
compounds as model substances like benzene, toluene, or
phenanthrene (e.g. Gustafsson et al., 1997; Kleineidam
et al., 2002; Zhu and Pignatello, 2005). The sorption of
these model substances on BC species has been shown to
be strong and highly non-linear (e.g. Accardi-Dey and
Gschwend, 2003; Braida et al., 2003; Nguyen et al., 2004;
James et al., 2005). Several attempts to explain the sorption
behaviour of organic materials with varying physicochem-
ical properties have been made during the last decade.
Chiou et al. (2000) introduced the term of ‘‘high surface
area carbonaceous material’’ (HSACM). Small amounts
of charred material like soot or charcoal are considered
to be responsible for the non-linear sorption of polar and
apolar components. The involved mechanisms are assumed
to be hydrophobic partitioning and pore filling, according
to the Polanyi–Manes (PM) model (Manes, 1998). The the-
ory postulates the existence of a nonpolar, microporous
adsorption site with a characteristic adsorption potential,
which is influenced by the nature of the sorbent, and the
distance of the sorption site from the sorbent surface. Also
interactions of water and adsorbent surface are taken into
account. The fundamental idea is a condensation of the
adsorbate in (micro-) pores while exhibiting the same phys-
ical properties as the unconfined bulk organic liquid or
solid. The sorption capacity is believed to be restricted by
the pore volume occupied by the mixed phase of condensed
liquids or solids. Consequently, the sorption of hydropho-
bic organic contaminants (HOC) on BC materials would be
mainly governed by its microporosity and its hydrophobic
character.
The physical properties of charcoals from burned bio-
mass are strongly dependent on the conditions during the
combustion process (Shafizadeh, 1984). According to
Schmidt and Noack (2000), charcoals produced by vegeta-
tion wildfires consist mainly of several randomly orientated
stacks of graphitic sheets. Still, the structure is influenced
crucially by a number of combustion parameters. The main
factors are the fuel type, fuel load, fuel condition, weather
condition, substrate heterogeneity, fire intensity, and dura-
tion (Patterson et al., 1987). Karapanagioti et al. (2004)
stated that ‘‘. . .the charcoal particle subgroup of organic
matter is heterogeneous in nature’’, and therefore con-
cluded that different subgroups of charcoal have to be dis-
tinguished in order to optimize the predictions for the bulk
sorption behaviour.
So far, little attention has been directed to the heteroge-
neity within the subgroup of charcoals. From work of Rai-
son (1979) and Scott (1989) we know that elevated ground
temperatures during most wildfires may vary between
200 C and 500 C, with highest temperatures for scrub-
land wildfires ranging up to levels between 700 C and
1000 C. Despite these extreme temperature ranges, artifi-
cial charcoals as used in most sorption studies consisted
either of charcoals produced from one fixed, or only a
small range of temperatures (Sander and Pignatello,
2005; Zhu and Pignatello, 2005; Zhu et al., 2005). A study
by James et al. (2005), however, employed a range of differ-
ent combustion temperatures for three species of softwood.
Still, the sorptive behaviour of the range of natural char-
coals remains poorly understood and data is lacking for
charcoals produced from different parent materials at var-
ious temperatures.
The objectives of this study were (i) investigate the sorp-
tive properties of charcoals produced from hardwood and
herbaceous material at various temperatures representative
of wildfires, and (ii) compare the sorption behaviour of two
simple aromatic model compounds on the range of char-
coals in single and bi-sorbate systems.
2. Experimental section
2.1. Wood and grass charcoal production
Red Gum (Eucalyptus camadulensis) chips, as used for
gardening purposes, were purchased from a landscape sup-
plier, and a pure stand of Phalaris pasture grass (Phalaris
tuberosa) was cut from a meadow. Both materials were
air dried at 40 C. Thin pieces of Red Gum wood were
hand picked, the Phalaris grass was milled into coarse seg-
ments and the dust was separated by sieving (2 mm grid).
To allow a uniform combustion process, the materials were
stacked uncongested in porcelain crucibles with lids. The
filled crucibles were weighed and subsequently placed in a
muffle furnace. For the charcoals combusted at 250 C
and 450 C, the furnace was ramped from room tempera-
ture to the final temperature in 1 h. As a second stage,
the final temperature was held for 2 h for the Red Gum
wood. Due to the more combustible nature of the Phalaris
grass, the final temperature was held only for 1 h in this
case. The furnace was then switched off to allow the cruci-
bles to cool down to room temperature. The Red Gum
wood treated at 850 C was combusted for only 1 h after
reaching the final temperature, for the Phalaris grass the
time was reduced to half an hour. The charred material
was weighed and grinded to powder on a N.V. THEMA
disc-rotating mill for 3 min. In the following, the abbrevia-
tions R250, R450, and R850 are used for the Red Gum
charcoals, P250, P450 and P850 for the Phalaris charcoals.
2.2. Determination of charcoal properties
The continuous flow method at 77 K was employed for
quantification of adsorbed and desorbed N2, using a
QUANTACHROME QUANTASORB QS-13 Surface-
Area Particle-Size Analyzer and ultra high purity gaseous
nitrogen (99.999%, from BOC Gases). Surface areas for
all six utilized charcoals were calculated from three-point
 L.C. Bornemann et al. / Chemosphere 67 (2007) 1033–1042 1035

regression, based on the BET equation after Brunauer et al.

(1938). Additionally, the pore size distribution of R450 and

N2 BET-SSAe
R850 was evaluated using BET nitrogen adsorption tech-
nique at 77 K. The details of the method and uncertainties

(m2 g 1)
associated with the measurement have been published

605

371
34

20
8

4
elsewhere (Badalyan and Pendleton, 2003). Outgassing of
the charcoal was carried out at 300 C for 8 h at a back-
ground vacuum of 1 · 10 4 Pa, similar to that used for
charcoal samples by Braida et al. (2003). For the pore vol-

(mlliq g 1)

0.02219
ume evaluation, we employed aS-plot analysis, where the

0.2757
Vtotald
adsorption properties of a porous solid are compared with

–
–
–
those of a non-porous standard material exhibiting surface
chemistry similar to the test sample (Badalyan and Pendl-

(mlliq g 1)
eton, 2003). Moisture content was determined by compar-

0.00375
Vmicroc

0.2469
ison of sample mass before and after evacuation at 300 C.
We classified pore width (dp), according to IUPAC recom-

–
–
–
mendations as follows: micropores: dp < 2 nm, mesopores:
2 < dp < 50 nm, and macropores: dp > 50 nm. Pores, which

Sum DMb
were smaller than 2 nm were analyzed using the Horvath–
Kawazoe (H–K) method (Horvath and Kawazoe, 1983).

(%)
0.4
0.4
0.3
1.9
1.8
1.6
This method calculates the effective micropore size distri-
bution of slit-shape pores using the data from adsorption
isotherms. Using the H–K equation we calculated effective

Sum CMa
mean width of slit pores (dp), and correlated it with the
value of DV ads
Drslit
, where DVads is the incremental amount of

(%)
0.6
0.9
1.3
2.7
4.4
5.8
nitrogen adsorbed (converted to condensed liquid volume
of nitrogen) and Drslit is the corresponding incremental
width of slit pores. The individual and cumulative frequen-
(mg kg 1)

cies were calculated for a range of dp data. Further infor- 6 110

8 690
13 150
27 400
44 070
58 200
Sum

mation on the characteristics of these materials has

recently been published by Smernik et al. (2006) and Yu
et al. (2006).
For the measurement of the total cation contents by
(mg kg 1)

atomic absorption spectroscopy (AAS), 0.2 g of the

840
1 280
2 580
17 640
29 240
36 080

charred material was weighed into microwave vessels and

K

2 ml H2O2 (35%) and 5 ml HNO3 (70%) were added. The

samples were then digested, using a MILESTONE mls
1200 Digestion Unit (300 W, 5 min, 600 W, 5 min, 500 W,
(mg kg 1)

3 min). The samples were converted into 50 ml volumetric

140
320
660
5 380
7 660
12 660

flasks and made up to volume in Milli-Q water. The liquid

Na

was filtered through 0.45 lm filters and 0.5 ml of the sample

was diluted 20 times prior to AAS analysis. Charcoal prop-
Specific surface area after Brunauer et al. (1938).

erties and composition details are presented in Table 1.

(mg kg 1)
Selected char properties and composition details

280
390
510
2 360
3 680
4 720
Mg

Sum of dry matter (precursor material).

Sum of combusted matter (Charcoal).

2.3. Batch sorption

All experiments were conducted in 32 ml KIMAX

(mg kg 1)

Micropore-volume (<2 nm).

glass culture-tubes (Kimble Glass Inc. USA) with PTFE-

4 850
6 700
9 400
2 020
3 490
4 740

lined screw lids to minimize sorption effects on the con-

Ca

tainer. Aliquots of 32 mg of charcoal were weighed in the

Total pore volume.

reaction vials and hydrated with 20 ml Milli-Q water for

Red Gum 250 C
Red Gum 450 C
Red Gum 850 C
Red Gum 250 C
Red Gum 450 C
Red Gum 850 C

42 h in an end-over-end shaker. A pre-hydrating period

Sample name

of at least 20–24 h is required due to the hydrophobic nat-

ure of the charcoal (Zhu and Pignatello, 2005; Zhu et al.,
Table 1

2005). After hydrating, the test sorbate was added as a

b
c
d
e
a

stock solution in a methanol-carrier. Benzene, toluene, pro-

1036 L.C. Bornemann et al. / Chemosphere 67 (2007) 1033–1042
pyl-benzene, methanol, and dichloromethane with HPLC-
grade purity (>99.9%) were used in the experiments. For
practical reasons the sorbate concentrations were prepared
as ll l 1and converted into lg l 1 in the calculations. In the
single-sorbate system, for the materials treated at 250 C
and 450 C, the initial sorbate concentrations were cover-
ing 2.20–87.90 lg ml 1 for benzene, and 2.17–86.6 lg ml 1
for toluene. Due to the considerably higher sorption capac-
ity of the 850 C materials the concentration range was
expanded to 351.6 lg ml 1 for benzene and 346.4 lg ml 1
for toluene. In the bi-sorbate systems both sorbates were
added in equal amounts, the total sorbate concentration
therefore being doubled, compared to the single-sorbate
systems. In order to prevent varying solvent effects, metha-
nol was always kept at 1% by volume throughout all
experiments. Specifications regarding the maximum con-
centration of carrier-solvent avoiding interference with
the sorption process are inconsistent (e.g. Kleineidam
et al., 2002; Karapanagioti et al., 2004; Sander and Pigna-
tello, 2005). However, we found in preliminary experiments
that keeping the solvent concentration constant at 1% pro-
vided reproducible results. To avoid headspace (in order to
minimize volatilization losses), the vials were filled up to
their capacity with Milli-Q water. As charcoal in such small
concentrations suspended in water would hardly settle
down during centrifugation, 0.32 ml of 1 M CaCl2-solution
was added in order to support the flocculation. A final con-
centration of 0.01 M CaCl2 was also chosen by other
authors (Ahmad et al., 2001; Sander and Pignatello,
2005). The vials were placed in the end-over-end shaker
again to equilibrate for 24 h. According to Braida et al.
(2003), at low initial concentrations (8 lg ml 1), benzene
sorption reached an equilibrium after eight days, but
the vast majority of benzene (>95%) was already sorbed
within 24 h. For higher initial benzene concentrations
(1600 lg ml 1) the apparent equilibrium was shown to
be reached after 24 h. Kwon and Pignatello (2005) reported
that maximum sorption of benzene on the charcoal also
utilized by Braida et al. (2003) was achieved within 20 h.
From these findings, together with observations that diesel
soot uptake of PAHs occurs within hours to one or two
days (Bucheli and Gustafsson, 2000) we concluded that
equilibration times of 24 h would be appropriate for our
purposes. The samples were then centrifuged at 650 g for
a duration of 10 min. Subsequently, 5 ml of the superna-
tant were transferred to PTFE-lined extraction vials with
15 ml capacity and extracted with 2 ml dichloromethane.
Prior to extraction, propyl-benzene was added as an inter-
nal standard. The extraction tubes were then shaken by
hand vigorously and an aliquot of the solvent was trans-
ferred into PTFE-lined GC-vials. Analysis of the extracts
was performed on a PERKIN ELMER Auto System Gas
Chromatograph equipped with a flame ionization detector
(FID). The column was a DB-5 column with a length of
30 m, an internal diameter of 0.25 mm and a film thickness
of 0.25 lm. The injection temperature was 200 C with 1 ll
of sample injected. Helium was used as the carrier gas with
a constant pressure of 22 psi. The detector was heated to
250 C. The initial temperature of the oven was set to
35 C, held for 3 min, and then ramped to 80 C with a rate
of 5 C min 1.
3. Results
3.1. Charcoal properties
Charcoals combusted at 250 C and 450 C exhibited
relatively low surface areas as revealed by N2-BET surface
area measurement, whereas the surface areas of the high
temperature treated charcoals (850 C) were very high
(Table 1). The Red Gum charcoals exhibited a much bigger
surface area than the Phalaris charcoals at corresponding
temperatures. The ratio of the surface areas of the Red
Gum charcoals to the ones from Phalaris drops from 2 at
250 C to 1.7 and 1.6 at the higher combustion tempera-
tures. For both materials the outstanding increase of the
surface areas (by almost 20 times) occurred between
450 C and 850 C. This critical change was also docu-
mented by the pore size distribution of R450 and R850.
The total pore volume of the 850 C material was higher
by more than an order of magnitude than that for the
450 C material (Table 1). Furthermore, the fraction of the
micropores (pores <2 nm), vmicro [mlliq g 1], revealed a
microporosity of 90% for the 850 C material, whereas
the 450 C material did not exceed a microporosity of
about 17%. The diagram displaying the integral pore size
distribution indicated a relatively wide range of pore
widths, i.e. 0.5–300 nm, the steep slope at about about
1.2 nm pore diameter indicating the large contribution of
this pore size for the 450 C material (Fig. 1). On the con-
trary, the range of pore size distribution for the 850 C
material was very narrow (Yu et al., 2006). Here, the over-
all dominating pore sizes ranged from only 0.4–1 nm, with
insignificant participation of pores with a diameter >10 nm
(Fig. 1). Corresponding to the observed N2-BET surface
areas, the pore size distribution illustrates the extraordi-
nary increase of the inner surface area with decreasing
diameter of the pores.
The total cation content of the Phalaris charcoals
exceeds the one of the Red Gum charcoals by a factor
greater than four (Table 1). Differences in the cation con-
tents of grass and wood are also documented in literature.
Harmand et al. (2004) found a value of about 0.4% for the
sum of calcium, magnesium and potassium in Red Gum
wood (Eucalyptus camadulensis) from Cameroon. Consid-
ering the low proportion of sodium contributing to the
total cation content of our Red Gum charcoal (2–5%),
the findings of Harmand et al. (2004) are in line with our
data (Table 1). For a Phalaris species harvested between
July and October in Sweden, the sum of the four cations
also determined in our study was reported to be about
1.8% of the dry matter (Burval, 1997), and thus again in
good agreement with our data set (Table 1).
 L.C. Bornemann et al. / Chemosphere 67 (2007) 1033–1042 1037

0.30 0.03

0.20 0.02

Vtotal [mlliq g ] R450

Vtotal [mlliq g-1] R850

-1
0.10 0.01

R850
R450

0.00 0.00
0.1 1.0 10.0 100.0 1000.0
d p [nm]

Fig. 1. Integral pore size distribution of the charred material. Total pore volumes (vtotal) of R850 (ordinate on the left) and R450 (ordinate on the right),
plotted against the diameter of the pores (dp).

3.2. Influence of the combustion temperature on the and found Langmuir-type sorption isotherms for R850,
sorption process after the classification by Giles et al. (1960). Both, benzene
and toluene were sorbed by R850 to a much higher degree
Fundamental differences in the sorptive behaviour of than by the other materials. Comparing the respective coef-
benzene and toluene were found for the Red Gum materi- ficients of determination (R2) and especially the standard
als treated at different temperatures. In Fig. 2, we plotted errors of estimate (SEE), the isotherms from single and
the sorbed concentration of sorbate q [mg g 1] against bi-sorbate experiments on R850 showed considerably bet-
the final concentration in aqueous solution Cf [mg ml 1] ter fits to the Langmuir than to the Freundlich equation
(Table 2). The calculated maximum uptake (Smax) of each
sorbate in the bi-sorbate system was about half of the max-
1000 imum uptake in the single-sorbate systems, where benzene
a: Benzene and toluene were sorbed in comparable amounts (Table 2).
Sorption isotherms of R250 and R450 could successfully
100 be described with the model after Freundlich. For benzene,
R250 and R450 exhibited a comparable sorption, whereas
10 toluene expressed a slightly higher affinity to R450 com-
R250
R450 pared to R250 (Fig. 2 a and b). In contrast to R850, the
sorption of toluene on R450 and R250 was significantly
q [mg g ]

R850
-1

1 higher than that of benzene (Fig. 3 a and b; Table 2).

1000
b: Toluene Whereas toluene was hardly affected by competition from
benzene, the benzene uptake even increased in combination
100 with toluene on both materials.

10 R250
3.3. Influence of the precursor material on the sorption
R450
R850 process
1
0.00 0.02 0.04 0.06 0.08 0.10 0.12 0.14 The apparent difference between the Phalaris- and Red
-1
C f [mg ml ] Gum-derived materials was the lower sorption capacity
of P850 as compared to R850 (Table 2). For P850, the cal-
Fig. 2. Sorbed amount of sorbate q plotted against the final sorbate
concentration Cf. Single-sorbate sorption isotherms for (a) benzene and culated maximum uptake after the Langmuir equation
(b) toluene on R250, R450, and R850. Error bars represent standard resulted in a value of Smax = 153 mg g 1, whereas the cor-
deviation of duplicate samples (n = 2). Symbols may cover error bars. responding value for the Red Gum charcoal was as large as
 1038
Table 2
Results of model fits to single and bi-sorbate isotherms
Char type Temperature (C) Single-/bi-solute Sorbate Smaxa (mg g 1) KLb (ml mg 1) R2c (–) SEE d
(–) ne (–) KFf (mln mg1 n
g 1) R2g (–) SEE h
(–)

L.C. Bornemann et al. / Chemosphere 67 (2007) 1033–1042

Phalaris 850 Single Benzene 153 ± 5.6 245 ± 91 0.99 0.05 0.41 ± 0.05 452.5 ± 1.5 0.86 0.24
Redgum 850 Single Benzene 222 ± 3.0 491 ± 99 1.00 0.05 0.45 ± 0.06 981.7 ± 1.7 0.83 0.29
Redgum 850 Single Toluene 236 ± 13.4 326 ± 121 0.97 0.03 0.39 ± 0.10 689.6 ± 2.2 0.60 0.38
Redgum 850 Bi Benzene 114 ± 8.3 1367 ± 373 0.96 0.01 0.44 ± 0.07 979.0 ± 1.8 0.87 0.22
Redgum 850 Bi Toluene 113 ± 1.2 8872 ± 2895 1.00 0.01 0.50 ± 0.18 7206.1 ± 7.7 0.51 0.37
Redgum 450 Single Benzene – – – 0.51 ± 0.4 60.9 ± 1.2 0.96 0.07
Redgum 450 Single Toluene – – – 0.50 ± 0.01 150.4 ± 1.1 1.00 0.03
Redgum 450 Bi Benzene – – – 0.48 ± 0.04 69.2 ± 1.2 0.96 0.07
Redgum 450 Bi Toluene – – – 0.47 ± 0.01 126.7 ± 1.1 0.99 0.04
Phalaris 250 Single Benzene – – – 0.56 ± 0.03 56.6 ± 1.1 0.98 0.05
Redgum 250 Single Benzene – – – 0.55 ± 0.04 63.1 ± 1.2 0.96 0.07
Redgum 250 Single Toluene – – – 0.58 ± 0.03 126.3 ± 1.1 0.98 0.05
Redgum 250 Bi Benzene – – – 0.56 ± 0.06 83.5 ± 1.3 0.92 0.10
Redgum 250 Bi Toluene – – – 0.52 ± 0.04 110.3 ± 1.2 0.95 0.08
Number of observations is 22 for benzene and toluene on materials treated at 850 C in single sorbate systems, 18 for all other systems.
a
Maximum sorbate uptake (Langmuir model).
b
Langmuir affinity coefficient.
c
Coefficient of determination (Langmuir model).
d
Standard error of estimate (Langmuir model).
e
Freundlich exponent.
f
Freundlich affinity coefficient.
g
Coefficient of determination (Freundlich model).
h
Standard error of estimate (Freundlich model).
 L.C. Bornemann et al. / Chemosphere 67 (2007) 1033–1042
100
a: R250
10
q [mg g-1]
1 (Fig. 1) is displaying the beginning of micropore formation.
100
b: R450
10
1 (2000) and indicate the pore filling mechanism. According
0.0001 0.001 0.01 0.1
C f [mg ml-1]
Benzene single Toluene single
Benzene bi Toluene bi
Fig. 3. Sorbed amount of sorbate q plotted against the final sorbate
concentration Cf. Sorption isotherms for (a) single- and bi-sorbate systems
of benzene and toluene on R250, and (b) single- and bi-sorbate systems of
benzene and toluene on R450. Error bars represent standard deviation of
duplicate samples (n = 2). Symbols may cover error bars (Freundlich-
linearization).
Smax = 222 mg g 1. Similar observations were made for the
250 C materials. The Freundlich sorption coefficient (KF)
for P250 showed reduced uptake of benzene compared to
R250 (Table 2), however, the slopes indicated by the Fre-
undlich exponent (n) were similar for both materials.
4. Discussion
4.1. Effect of the combustion temperature on sorption
properties of charcoal
Except for R850, surface areas of laboratory-produced
charcoals in published studies are generally comparable
with our data. The surface area of 600 m2 g 1and a
microporosity of 90% as observed for R850 exceeds the
maximum values found in the literature. According to San-
der and Pignatello (2005), N2-BET surface areas for typical
charcoals ranged from 200–500 m2 g 1. However, litera-
ture data (Nguyen et al., 2004; James et al., 2005) and
our own results suggest that surface areas of charcoals
may change considerably with varying combustion temper-
atures and precursor materials. Surface areas and the pore
volumes appear to escalate, and the pore size distribution
shifts to a mostly microporous pattern above a critical tem-
perature. According to Hellwig (1985), combustion temper-
atures of 250 C are only sufficient to cause the evolution of
volatiles, whereas at temperatures as high as 450 C the
softening of the woody material is already initiated. Results
of Haghseresht et al. (1999), who examined carbonaceous
1039
adsorbents using 13C NMR, indicated that increased heat
treatment results in enhanced contribution of aromatic
structures and an increase of amorphous carbon and
molecular disorder, thereby having impact on amount
and size of micropores. Although the amount of microp-
ores is still low, the maximum peak for the pore widths
of about 1.1 nm for the material combusted at 450 C
It is noteworthy that despite a significantly higher hydro-
phobicity of toluene (benzene: KOW = 134.9; toluene:
KOW = 498.8), the calculated maximum uptake (Smax) on
R850 was in about the same order of magnitude for both
sorbates in the single-sorbate systems, as well as in the bi-
sorbate system. Especially the results of the bi-sorbate
experiments are consistent with the theory of Chiou et al.
to the assumption of an universal adsorbate for the pore fill-
ing process, the amount of adsorbed benzene and toluene
should be about the same when identical volumes of the sor-
bates are added. Furthermore, the suppression of the
sorbed amount of benzene and toluene from the respective
competitor in the bi-sorbate system compared to the single-
sorbate system should be about 50% and almost identical
for both sorbates, since the densities of benzene and toluene
are differing only slightly (benzene: 0.879 g ml 1; toluene:
0.866 g ml 1). As seen in Table 2, the calculated maximum
uptake for benzene and toluene in the bi-sorbate experiment
was indeed identical for each sorbate and about the half of
the calculated capacities in the single-sorbate experiments.
Considering that in the bi-sorbate system the two sorbates
were added in identical amounts, condensation of the sor-
bates in the pores of this highly microporous material
(Table 1) could explain the observed phenomenon. This is
also supported by studies of Zhu and Pignatello (2005),
addressing pore filling as the most likely dominant process
for the sorption of aromatic compounds on such materials.
In their study, the sorptive abilities of a charcoal from
maple wood shavings were compared with that of (non-por-
ous) graphite as a model. The experiments displayed size
exclusion effects on the charcoal for compounds exceeding
a certain molecular size, and enhanced sorption on the char-
coal compared to the graphite, despite a charcoal graphite
partition coefficient >1. By investigating the effect on pore
blocking by lipids on the same material as used by Zhu
and Pignatello (2005), Kwon and Pignatello (2005) showed
that benzene sorption is largely located in the interior pore
work. Still, essential differences are apparent between the
material used by Zhu and Pignatello (2005) and the char-
coals used in our study. The surface area of the maple wood
charcoal (400 m2 g 1) examined by Zhu and Pignatello
(2005) was considerably lower than the one of our R850
material (605 m2 g 1). Also the microporosity of our mate-
rial (90%, Table 1) exceeds the one of the char from maple
wood shavings (80%). The degree of non-linearity of the
sorption process increases with increasing microporosity
(after the Polanyi–Manes model), and might thus explain
that the isotherms for our R850 could be described by the
1040 L.C. Bornemann et al. / Chemosphere 67 (2007) 1033–1042
Langmuir model, whereas the isotherms for the maple
wood char showed better fit to a Freundlich sorption iso-
therm. The sorption on charcoals produced at lower tem-
perature in our study also fitted better to Freundlich model.
Kleineidam et al. (2002) investigated the sorption of dif-
ferent low-polarity organic compounds on a series of carbo-
naceous adsorbents, covering humic soil organic matter,
thermally altered carbon materials, and purely engineered
microporous sorbents. Employing the PM-model combined
with linear partitioning for sorbents containing humic
material, they were able to predict unique sorption iso-
therms by employment of solubility normalized aqueous
concentrations. Their results suggest that the PM-model is
the most appropriate model for sorption processes on
highly porous materials. However, some concerns about
potential artefacts resulting from enhanced water adsorp-
tion to functional groups of sorbents with varying hydro-
philicity have been expressed (Li et al., 2005). Also recent
results by Yang et al. (2006) showed that gaps in the under-
standing of the PM-model still exist. Their study revealed,
that the molar volume alone (as employed e.g. by Sander
and Pignatello, 2005) in some cases may fail to be an appro-
priate scaling factor in order to obtain matching isotherms
of hydrophobic compounds with varying molecular sizes.
Although the PM-model is increasingly used for modelling
of sorption processes, our results among with those of other
researchers (e.g. Xia and Ball, 2000) show that the classical
approaches like the Freundlich model and the Langmuir
model may still be adequate in some cases. These models
are appropriate here, as the aim of our investigations was
to detect general differences in the sorptive properties of
various charcoals, rather than seeking a specific mechanism
or an improvement in modelling of sorption isotherms.
The higher sorption of toluene in comparison with ben-
zene on R250 and R450 (Fig. 3a and b; Table 2) is possibly
due to its higher hydrophobicity. However, toluene sorption
in single and bi-sorbate system was the same, showing no
competitive effect. Sander and Pignatello (2005) examined
the competitive sorption of benzene, toluene, and nitroben-
zene on the charcoal mentioned above, which was also uti-
lized by Zhu and Pignatello (2005). In their bi-sorbate
experiments, the amount of competitive sorbate was kept
constant while the concentration of the test sorbate chan-
ged. From their resulting competitive sorption isotherms it
can be seen that competition effects were most pronounced
when the competitor solutes were added at high concentra-
tions together with a low concentration of test sorbate.
Competition effects diminished as the concentrations of
the two competing sorbates converged. This explains the
lack of significant competition effect from either sorbate in
our bi-sorbate systems, as in our experiments both sorbates
were always added in equal concentrations.
In our bi-sorbate experiments, the sorption of benzene on
R250 and R450 was even enhanced compared to the single-
sorbate system (Fig. 3a and b; Table 2). The reasons for this
are unclear, but may be related to the ability of aromatic
compounds like benzene and toluene to cause the swelling
of charcoal particles. Jonker and Koelmans (2002) found
in their experiments that different solvents exhibited varying
abilities to extract PAHs from soot and sediments and pro-
posed a two-step mechanism. According to them, the PAHs
are being extracted by an initial swelling of the sorbent
matrix with a subsequent displacement of sorbates by sol-
vent molecules. Among a selection of seven popular sol-
vents, also comprising two mixtures of benzene and other
solvents, toluene was proven to be the best suited solvent
for the extraction of PAHs from charcoal. Therefore, the
enhanced sorption of benzene in the bi-sorbate systems
may result from a toluene-induced swelling of the matrix,
thereby opening up additional sorption sites for the smaller
benzene molecules and toluene itself being sterically hin-
dered to penetrate by its larger molecular size. Braida
et al. (2003) proved that certain benzene concentrations
were able to induce the swelling of charcoal matrices,
thereby increasing the sorption capacity and inducing sorp-
tion hysteresis by pore deformation and solvent entrapment.
4.2. Influence of the precursor material on the sorption
process
The lower (33%) sorption capacity of P850 compared
to R850 (Table 2) is ascribed to the physicochemical differ-
ences between the respective materials. Although the expo-
sure time to the heat treatment was already reduced due to
the more combustible nature of the grass, the charred grass
appeared to be much more disaggregated compared to the
wood charcoal. A more extensive combustion, resulting in
the loss of sorption active constituents may have resulted in
the reduced sorption capacity. James et al. (2005) found
that surface area, pore volume, and microporosity are
not increasing endlessly with increasing combustion tem-
peratures, and that softening temperatures are also depen-
dent on the precursor material. In their study, a heating
temperature of 820 C resulted in drastically reduced sur-
face areas and microporosity for a charcoal from Betula
pendula, compared to charcoal produced at a temperature
of 700 C. For Auraucaria araucana, the softening temper-
ature was already reached at 450 C. Another explanation
for the reduced sorption capacities could be the overall
higher cation content in Phalaris grass compared to Red
Gum wood (Table 1). As the charcoal was weighed in as
bulk, and was not corrected for its content of mineral con-
stituents, a higher cation content reduces not only the pro-
portion of highly sorptive BC in the bulk, but also increases
the hydrophilic character of the whole sample. The latter is
true as these cations are present as salts, their hydrophilic
character hindering the sorption of hydrophobic compo-
nents. Haghseresht et al. (1999) found that the content of
aromatic carbon in the precursor correlated with the con-
tribution of amorphous carbon in the corresponding char-
coal. A high amount of amorphous carbon results in an
increased abundance of sorption-active edge sites as well
as in an increased disorder, thereby supporting the forma-
tion of micropores and increasing the sorption capacity
 L.C. Bornemann et al. / Chemosphere 67 (2007) 1033–1042
(Haghseresht et al., 1999). The content of aromatic carbon
in plant tissue is mainly governed by its lignin content. The
lignin content of hardwood (35%) (Anderson and Till-
mann, 1977) is considerably higher than the one for
grasses, with values ranking between 3% and 10% (But-
ler and Bailey, 1973). We suggest that also the slightly
lower sorption capacity of Phalaris charcoals is likely to
be the combined effect of their smaller surface area, stron-
ger hydrophilic effects from mineral salts, and lower lignin
content.
5. Conclusions
The results from the present studies demonstrated that
charcoals prepared under different combustion tempera-
tures exhibit substantially different physicochemical and
thus also sorptive properties, especially if a critical ‘‘soften-
ing temperature’’ is exceeded during combustion. For the
observed precursor materials, this critical temperature
was within the scope of temperatures observed for wildfires
(200–1000 C) (Raison, 1979; Scott, 1989). The charcoals
derived from herbaceous material expressed basically com-
parable sorptive patterns to wood charcoals. However,
their mineral contents were higher, whereas the N2-surface
areas and sorption capacities were lower than for the wood
charcoals. The observed highly non-linear sorption behav-
iour of R850 and P850 showed better fits to Langmuir iso-
therms than to Freundlich isotherms and was most likely
indicative of pore filling processes on the highly micropo-
rous charcoal studied here. The surface area varied orderly
with increasing combustion temperature among the
observed charcoals and gave a good indication for the
sorptive abilities of the charcoals. Also other parameters
such as hydrophobicity and swelling of the sorbing matrix
seemed to be important for sorption of aromatic hydrocar-
bons, as indicated by the enhanced sorption of benzene on
R250, R450, and P250 in the presence of toluene. More
work is needed to establish the effect of combustion condi-
tions on the nature of charcoals and their subsequent
impact on sorption and desorption behaviour of organic
contaminants.
Acknowledgements
The measurement of pore size distributions was con-
ducted by Dr. Alexander Badalyan (University of South
Australia), N2 BET-SSA and total cation contents were
determined by Mr Lester Smith (CSIRO Australia). Ms
Natasha Waller (CSIRO) provided valuable technical assis-
tance during these studies. The authors would like to thank
Dr. Sonja Brodowski (University of Bonn) and Dr. G.-G.
Ying (CSIRO) for their comments on the manuscript. Lud-
ger Bornemann was partially funded by the ‘‘Studienstif-
tung des deutschen Volkes’’ while working on this project
at CSIRO laboratories in Adelaide.
1041
Appendix A
Table of symbols
Cf final sorbate concentration (mg ml 1)
Kd soil water partition coefficient (l kg 1)
KF Freundlich affinity coefficient (mln
mg1 n g 1)
KL Langmuir affinity coefficient
n Freundlich exponent
q sorbed amount of sorbate per mass of
sorbent (mg g 1)
Smax maximum sorbate uptake (mg kg 1)
(Langmuir model)
SW water solubility (mg l 1)
V total volume (ml)
vmicro volume of micropores (mlliq g 1)
VP vapour pressure (mm HG at 20 C)
vtotal total pore volume (mlliq g 1)
dp pore diameter (nm)
DVads incremental amount of nitrogen absorbed
(mlliq g 1)
Drslit incremental width of slit pores (nm)
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Sorption and Desorption Behaviors of Diuron in Soils Amended

with Charcoal
XIANG-YANG YU,†,‡ GUANG-GUO YING,†,§ AND RAI S. KOOKANA*,†
CSIRO Land and Water, Adelaide Laboratory, PMB 2, Glen Osmond 5064, South Australia,
Australia; Food Safety Research Institute, Jiangsu Academy of Agricultural Sciences,
Nanjing 210014, China; and Guangzhou Institute of Geochemistry, Chinese Academy of Sciences,
Guangzhou 510640, China

Charcoal derived from the partial combustion of vegetation is ubiquitous in soils and sediments and
can potentially sequester organic contaminants. To examine the role of charcoal in the sorption and
desorption behaviors of diuron pesticide in soil, synthetic charcoals were produced through
Published on October 11, 2006 on http://pubs.acs.org | doi: 10.1021/jf061354y

carbonization of red gum (Eucalyptus spp.) wood chips at 450 and 850 °C (referred to as charcoals
BC450 and BC850, respectively, in this paper). Pore size distribution analyses revealed that BC850
contained mainly micropores (pores ≈ 0.49 nm mean width), whereas BC450 was essentially not a
Downloaded by UNIV OF GEORGIA on August 4, 2009

microporous material. Short-term equilibration (<24 h) tests were conducted to measure sorption
and desorption of diuron in a soil amended with various amounts of charcoals of both types. The
sorption coefficients, isotherm nonlinearity, and apparent sorption-desorption hysteresis markedly
increased with increasing content of charcoal in the soil, more prominently in the case of BC850,
presumably due to the presence of micropores and its relatively higher specific surface area. The
degree of apparent sorption-desorption hystersis (hysteresis index) showed a good correlation with
the micropore volume of the charcoal-amended soils. This study indicates that the presence of small
amounts of charcoal produced at high temperatures (e.g., interior of wood logs during a fire) in soil
can have a marked effect on the release behavior of organic compounds. Mechanisms of this apparent
hysteretic behavior need to be further investigated.

KEYWORDS: Diuron; sorption; desorption; soil; charcoal; carbonization; hysteresis

INTRODUCTION
Black carbon (e.g., soot and charcoal) produced from the
incomplete combustion of vegetation and fossil fuel is ubiquitous
in terrestrial and aquatic environments (1, 2). In addition to
dispersal through biomass and fossil fuel combustion in the
environment, some agricultural practices may also contribute
to the increasing amount of black carbon in agricultural soils.
For example, it is an old practice in the eastern and southern
parts of China to mix firewood ashes with soils and livestock
dung followed by heating and aging for several months, before
the mixture is added directly into the field as a fertilizer. Direct
burning of plant residues in the field after harvest for land
clearing is common all over the world. Such agricultural
practices will also provide direct input of black carbon into
agricultural soil (3-6). Terrestrial black carbon, being erosion-
prone, is readily transported by wind and water and is often
deposited into aquatic ecosystems. Black carbon has been found
to make a significant proportion of soils and sediments: 15-
* Corresponding author (e-mail rai.kookana@csiro.au; fax +61 8
83038565).
† CSIRO Land and Water.
‡ Jiangsu Academy of Agricultural Sciences.
§ Chinese Academy of Sciences.
10.1021/jf061354y CCC: $33.50 © 2006 American Chemical Society
Published on Web 10/11/2006
30% of total organic matter in marine sediments (7) and 12-
31% of deep-sea sediments (8), 18-41% of the soils and
sediments collected from the suburban area of Guangzhou,
China (9), up to 30% of soils collected around Australia (4,
10), and up to 45% of total organic carbon (TOC) in soils
collected from Germany (11).
It has been well recognized that the presence of charcoal in
soil could not only enhance the sorption of organic contaminants
such as pesticides (12) but also influence the nature of the
sorption mechanism (13). Research in recent years (14) has
shown that the residues produced from burning wheat and rice
were 400-2500 times more effective than soil in sorbing diuron
over the concentration range of 0-6 mg/L in water. However,
the effect of these amendments on the desorption behavior of
diuron was not investigated in that study.
The nature and properties of black carbon are strongly
affected by the nature of parent materials (wood, grass) and
the temperature experienced during combustion (2, 15). Char-
coals formed from lignocellulosic materials under conditions
of pyrolysis become increasingly carbonized as temperatures
rise above 500 °C and are completely carbonized as temperatures
approach 1000 °C (15). Under relatively high pyrolysis tem-
peratures (500-700 °C), the charcoal derived from wheat was
 8546 J. Agric. Food Chem., Vol. 54, No. 22, 2006
found to be well carbonized and had a relatively high surface
area and low oxygen content (16). Such chars have been reported
to show high affinity for organic compounds (16, 17), especially
for polar solutes (16).
Despite the fact that the environmental fate and impact of
contaminants (bioaccessibility, bioavailability, and toxicological
impact) are strongly influenced by their desorption behavior
(18-21), the effect of black carbon on the desorption behavior
of compounds has so far received limited attention. Braida et
al. (22) observed pronounced hysteresis in the sorption of
benzene in water to pure-form maple-wood charcoal prepared
by oxygen-limited pyrolysis. Here they found that the sorption
of wood charcoal was highly irreversible and proposed that
hysteresis was due to pore deformation of charcoal during
desorption of the sorbate, leading to entrapment of molecules
as the polyaromatic scaffolding collapsed during desorption.
The objective of this study was to assess the effect of the
presence of small amounts of charcoal materials (prepared from
eucalyptus wood at two different temperatures of 450 and
850 °C) in soil on the sorption-desorption behaviors of pesti-
cides in soil. Sorption and desorption behaviors were investi-
Published on October 11, 2006 on http://pubs.acs.org | doi: 10.1021/jf061354y
gated to assess the role of the types and amounts of charcoal as
sorbent for pesticides (represented by diuron as a model nonionic
pesticide) in soils amended with various amounts of charcoals.
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EXPERIMENTAL PROCEDURES
Chemicals. Diuron [N-(3,4-dichlorophenyl)-N,N-dimethyl urea, with
purity >99%] was obtained from Sigma-Aldrich (Sydney, Australia).
Diuron is a nonionic urea herbicide with a very wide use globally.
The herbicide is nonvolatile (vapor pressure is 1.1 × 10-3 mPa at
25 °C), has a water solubility of 36.4 mg/L, and is stable in neutral
media. Its reported average soil half-life is in the range of 90-180
days (23). Sodium azide was obtained from Fluka (Sydney, Australia).
Calcium chloride of analytical grade and all solvents of HPLC grade
were obtained from Merck Pty. Ltd. (Victoria, Australia). A stock
solution of 100 mg/L of diuron was prepared in methanol.
Charcoal, Soil, and Sorbent. Charcoals were prepared from red
gum wood (Eucalyptus spp.), at two different temperatures, namely,
450 and 850 °C (BC450 and BC850, respectively). Red gum woodchips
were air-dried at 40 °C, and pieces of <5 mm were hand picked for
the carbonization process. The woodchips were placed in porcelain
crucibles with lids in a temperature-programmable muffle furnace
(S.E.M., Australia). The temperature of the furnace was ramped to the
defined temperature (450 or 850 °C) and held for 2 h for BC450 and
for only 1 h for BC850. The prepared black carbon materials were
ground to a fine powder on a disk-rotating mill for 3 min.
The specific surface areas (SSA) of the two charcoals were evaluated
using the Brunauer, Emmett, and Teller (BET) nitrogen adsorption tech-
nique (24) at 77 K, using an automated manometric gas adsorption
apparatus (25) and ultrahigh-purity gaseous nitrogen (99.999%, from
BOC Gases). The details of the method and uncertainties associated
with the measurement have been published elsewhere (25). Outgassing
of the charcoal was carried out at 300 °C for 8 h at a background
vacuum of 1 × 10-4 Pa, similar to that used for charcoal samples by
Braida et al. (22). For the pore volume evaluation, we employed RS-
plot analysis, where the adsorption properties of a porous solid are
compared with those of a nonporous standard material exhibiting surface
chemistry similar to that of the test sample (25). Moisture content was
determined by comparison of sample mass before and after evacuation
at 300 °C.
We classified pore width (dp) according to the International Union
of Pure and Applied Chemistry (IUPAC) recommendations as fol-
lows: micropores, dp < 2 nm; mesopores, 2 < dp < 50 nm; and macro-
pores, dp > 50 nm. Pores smaller than 2 nm were analyzed using the
Horvath-Kawazoe (H-K) method (26). This method calculates the
effective micropore size distribution of slit-shape pores using the data
from adsorption isotherms. Using the H-K equation we calculated the
effective mean width of slit pores (dp) and correlated it with the
Yu et al.
value of ∆Vads/∆rslit, where ∆Vads is the incremental amount of nitrogen
adsorbed (converted to condensed liquid volume of nitrogen) and ∆rslit
is the corresponding incremental width of slit pores. The individual
and cumulative frequencies were calculated for a range of dp data. Other
properties of these chars as characterized using NMR techniques have
been published elsewhere (27).
The soil used in this experiment was collected from the Roseworthy
Campus, University of Adelaide, and was amended with charcoal. After
air-drying, the soil was passed through a 2 mm sieve. This soil contained
87.8% sand, 1.3% silt, 8.3% clay, and 1.4% organic matter and is of
sandy loam texture. The soil pH value was 6.8 in a 1:5 (soil/water)
soil suspension and had a maximum water-holding capacity of 34.2%
(v/v) and a cation exchange capacity of 9.3 cmol(+)/kg. The soil was
sterilized by autoclaving at 120 °C under 300 kPa chamber pressure
for 30 min three times within 3 days.
Charcoal-amended soils used in the experiment were prepared by
mixing the above soil and the two types of charcoal at different ratios.
The percentages of charcoal materials in the amended soils were 0,
0.1, 0.5, 1.0, 2.0, and 5.0% (w/w) for BC450 and 0, 0.1, 0.2, 0.5, 0.8,
and 1.0% (w/w) for BC850. The charcoal-amended soils were thor-
oughly mixed on a rotary shaker for 7 days before their use as sorbents
for sorption and desorption experiments.
Sorption and Desorption Isotherms. Diuron sorption by the
sorbents was measured by the batch equilibration technique. Preliminary
kinetic experiments showed sorption and desorption of diuron in
charcoal-amended soils reached an apparent equilibrium within 24 h.
Although sorption is known to continue for days at a very slow rate,
for the comparative assessment between different chars under similar
conditions, the 24 h shaking time was deemed to be adequate for the
purposes of this study. The sorbents were suspended in 10 mL of 0.01
M CaCl2 solutions (containing 0.5% NaN3 to inhibit microbial activity)
spiked at concentrations from 1 to 27 mg/L of diuron. The amounts of
charcoal-amended soil used in the experiments were adjusted to allow
for 30-80% of the added chemical to be sorbed at equilibrium. On
the basis of our preliminary experiments, for BC450-amended soil an
aliquot of 1.0 g of soil was used for 0, 0.1, 0.5, 1.0, and 2.0% and only
0.2 g of soil for 5.0%. For BC850-amended soils, an aliquot of 0.5 g
of soil was used for 0.1 and 0.2% and only 0.2 g for 0.5, 0.8, and
1.0% amendment. The suspensions were shaken on a rotary shaker at
room temperature (22 ( 2 °C) at 120 rpm for 24 h and then centrifuged
at 4000 rpm for 60 min. After centrifugation, an aliquot of the super-
natant in each tube was taken out and analyzed directly by high-
performance liquid chromatography (HPLC).
Each sorption test was carried out in triplicate. Losses during the
test were monitored by including two blank controls in each test: one
tube that had only a chemical solution without any sorbent and the
other control tube that had only the sorbent and CaCl2 solution without
chemical. Tests showed losses due to adsorption to glassware and
degradation were negligible, and no interferences were found during
the analysis of solutions. This is consistent with previous studies and
the nature of the chemical (persistent and nonvolatile).
Desorption experiments were conducted for those samples with the
highest sorption loading. After 24 h of equilibration, the tubes were
centrifuged and 5 mL of the supernatant in each tube was taken out
for analysis. Another 5 mL of 0.01 M CaCl2 (including 0.5% NaN3)
was added into each tube, and the tubes were shaken for 24 h again.
The desorption process was repeated three more times for each tube.
All tests were performed in triplicate.
Pesticide Analysis. Analysis of diuron in the supernatant fraction
from sorption and desorption experiments was carried out on an Agilent
1100 series high-performance liquid chromatograph (HPLC) fitted with
a diode array detector and an SGE C18 RS column (250 × 4.6 mm,
5 µm). Acetonitrile (ACN) and water were used as the mobile phase,
which was programmed from 36% ACN at 0 min to 80% ACN at 6 min
at a flow rate of 1 mL/min. The UV wavelength for detection of diuron
was 248 nm. The detection limit for diuron was 0.03 mg/L.
Data Analysis. The sorption and desorption isotherms were fitted
with the linear form of the Freundlich equation
log Cs ) log Kf + N log Cw
 Sorption/Desorption of Diuron in Charcoal-Amended Soils J. Agric. Food Chem., Vol. 54, No. 22, 2006 8547

where Cw is the concentration of the chemical in solution, Cs is the

concentration of the chemical in the sorbent, Kf is the Freundlich
sorption coefficient, and N is the exponent indicative of sorption
mechanism. The sorption-desorption apparent hysteresis index (H) was
determined by the equation (28)

H ) N/Nd

where N and Nd are the Freundlich exponents calculated from the

sorption and desorption isotherms, respectively.

RESULTS AND DISCUSSION

Characteristics of Charcoals. Upon carbonization at 450
and 850 °C, 100 g of red gum chips produced about 36.1 and
28.8 g of charred materials, respectively. These are relatively
higher yields of charcoal than those derived from burning crop
residues (29). The color was gray to black for the BC850,
whereas for BC450 it was a little brown to black. The relatively
lower mass that remained under higher burning temperature
indicates that BC850 is carbonized to a greater degree than
BC450. The higher mass recovery of red gum chip-derived
Published on October 11, 2006 on http://pubs.acs.org | doi: 10.1021/jf061354y

charcoal is believed to be due to the more condensed structure

of wood chips than that of crop residues.
For the charcoal BC450, the shape of the nitrogen adsorption
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isotherm was similar to that for nonporous materials (Figure

1A), suggesting a negligibly small micropore volume (30). In
contrast, the BC850 is a microporous material with a high
specific surface area. The initial steep portion of the adsorption
isotherm for BC850 is indicative of the presence of micropores
in the charcoal material (30).
The SSA for BC850 was much higher than that of charcoal
BC450 (Table 1) and also higher than wheat-residue-derived
chars, the highest SSA of which was reported to be 438 m2/g
(charred at 600 °C) (16). Chun et al. (16) also found char surface
area to increase with increasing charring temperatures (300-
600 °C); however, SSA measurement of char produced at
700 °C was lower than that of char produced at 600 °C, which
they speculated to be due to microporous structures which were
destroyed at 700 °C.
Charcoal BC450 has a very low level of microporosity (peak
maxima occurred at a pore width of about 1.1 nm) (Figure 2).
This may indicate the beginning of micropore formation at
450 °C. Increasing the preparation temperature, as in the case Figure 1. Nitrogen adsorption/desorption isotherm for (A) BC450 and
of charcoal BC850, clearly promoted the formation of micro- (B) BC850.
pores with the maximum peak occurring for pore widths of about
0.49 nm and essentially all pores <1 nm in pore width. Table 1. Characteristics of Black Carbon Materialsa
From the pore volume values listed in Table 1, we regard
charcoal BC450 as a non-microporous material. Therefore, the BC
lower temperature (450 °C) used for the preparation of charcoal burning
temp SSA
moisture
content
micro
V total pore
V total
materials did not lead to the formation of microporous structure. (°C) abbrev (m2/g) (%) [mL (liq)/g] [mL (liq)/g]
In contrast, charcoal BC850 is predominantly a microporous 450 BC450 27.330.035 11.7 0.00374 ± 0.00004 0.02219 ± 0.00004
material with about 89.6% of volume from micropores (in terms 850 BC850 566.390.31 9.9 0.2469 ± 0.0039 0.2757 ± 0.0015
of specific micropore volume). It is customary to express the
a V micro ) volume of total specific microspore; V pore ) volume of specific
total specific pore volume (V pore
total) of an adsorbent as the liquid total total
volume adsorbed at a certain value of relative pressure (P/P0 microspore and total specific pore.
) 0.95) (31). The gaseous nitrogen volume adsorbed at this
value of relative pressure was converted into a liquid volume lich equation (Table 2); however, at the highest level of charcoal
of nitrogen. For mixed-pore materials, the mesopore volume is amendment, the fit was noted to be relatively poor in both types
determined as the difference between the total specific pore of chars. Instead, the data showed a much better fit to the
volume and total specific micropore volume. Langmuir model in these cases. The increasing Freundlich
Sorption-Desorption Isotherms and Their Nonlinearity. sorption coefficient Kf values and decreasing N values with
The presence of charcoal in the soil caused the sorption iso- charcoal content in soil (Table 2) show that the sorption capacity
therms to change progressively into highly nonlinear, concave- of diuron on charcoal-amended soils gradually increased with
downward-shaped isotherms (Figures 3 and 4). Most of the data increasing content of charcoal in soil. The N values decreased
on sorption and desorption isotherms fitted well to the Freund- from 0.83 to 0.25 and 0.16, respectively, for BC450 and BC850,
 8548 J. Agric. Food Chem., Vol. 54, No. 22, 2006 Yu et al.

Figure 3. Influence of BC450 on sorption and desorption of diuron in

soil. (Inset) Sorption−desorption on unamended soil and on 0.1% BC450.
Published on October 11, 2006 on http://pubs.acs.org | doi: 10.1021/jf061354y
Downloaded by UNIV OF GEORGIA on August 4, 2009

Figure 4. Influence of BC850 on sorption and desorption of diuron in

soil. (Inset) Sorption−desorption on unamended soil and on 0.1% BC850.

Table 2. Freundlich Constants for the Sorption (Kf, N) and Desorption

(Kfd, Nd) of Diuron on Soil Amended with Charcoal and the Calculated
Hysteresis Index (H)
content of
charcoal in
charcoal soil (%) Kf N R2 Kfd Nd R2 H
original soil 4.08 0.83 0.9974 5.40 0.73 0.9975 1.14
BC450 0.1 4.47 0.82 0.9994 11.1 0.48 0.9944 1.69
Figure 2. Pore size distribution for (A) BC450 and (B) BC850. Using the
0.5 28.0 0.45 0.9983 47.5 0.23 0.9990 1.93
Horvath−Kawazoe (H−K) method (26), we calculated the effective mean 1.0 514 0.37 0.9979 78.6 0.21 0.9977 1.79
width of slit pores, dp, and plotted against the value of ∆Vads/∆rslit, where 2.0 109 0.34 0.9958 168 0.12 0.9772 2.87
∆Vads is the incremental amount of nitrogen adsorbed (converted to 5.0 314 0.25 0.9980 427 0.10 0.8784 2.37
condensed liquid volume) and ∆rslit is the corresponding incremental width BC850 0.1 25.3 0.37 0.9979 39.5 0.28 0.9958 1.32
of slit pores. 0.2 72.8 0.32 0.9911 138 0.09 0.9926 3.54
0.5 224 0.22 0.9875 371 0.04 0.9990 5.14
0.8 350 0.21 0.9928 576 0.02 0.9004 9.75
as the amount of charcoal in soil increased from 0.1 to 5.0% of 1.0 500 0.16 0.9249 727 0.01 0.8194 14.92
BC450 and to 1.0% of BC850 (Table 2).
Highly nonlinear sorption isotherms and high sorption capac-
ity for organic compounds on sorbents (e.g., soils and sediments) of 1 mg/L, where Kf ) Kd(Cs/Cw)]. On this basis the sorption
containing black carbon have been observed in a number of capacities of the charcoal amended soil were 7-80 times for
studies (e.g., refs 32-35). Soil organic carbon has commonly BC450 and 5-125 times for BC850 in comparison to that of
been hypothesized to consist of an amorphous phase and a charcoal-free soil (Table 2). The sorption contribution of even
condensed phase of organic carbon (36, 37), which are consid- small additions of charcoals to the soil (0.5% of BC450 and
ered to be responsible for partitioning and sorption, respectively. 0.1% of BC850) was very high indeed (>85%), assuming no
Depending upon the relative contents of the two phases of change in inherent sorption capacity of soil. This shows that
organic carbon, sorption by soils and sediments could range the presence of even small amounts of highly carbonaceous
from linear partitioning to highly nonlinear sorption (34, 38, black carbon (charcoal, soot, or other carbonaceous materials)
39). can dominate sorption of an organic compound in soils and
Due to the highly nonlinear nature of isotherms, a comparison sediments.
of sorption affinities among the charcoal-amended soils needs Desorption Behavior and the Role of Microporosity. Sorp-
to be calculated for a specific solution concentration [e.g., Cw tion and desorption isotherms were compared to assess the
 Sorption/Desorption of Diuron in Charcoal-Amended Soils J. Agric. Food Chem., Vol. 54, No. 22, 2006 8549

degree of reversibility of sorption reaction. Due to the short

equilibration time (24 h) employed here, it is not appropriate
to refer to the discrepancy between the sorption and desorption
flanks of the isotherms as true hysteresis. This is because the
sorption kinetics may have been slow and the sorption reaction
may have continued beyond the point in time when the desorp-
tion part of experiment was initiated. Nevertheless, a comparison
of these isotherms does provide an assessment of the impact of
the nature of sorbate on the desorption behavior of diuron or
lack of it. For the purposes of this study, by hysteresis we mean
“apparent hysteresis”.
The highly nonlinear sorption isotherm and relatively flat
desorption isotherm in Figures 3 and 4 provide visual evidence
of apparent sorption-desorption hysteresis on the charcoal-
amended soils. We calculated the hysteresis index (H), defined
as the N/Nd ratio (28), to quantify the degree of apparent hys-
teresis. The H values for all of the sorbents are listed in Table
2. For the original soil sample the H value was 1.14, indicating
minimal desorption hysteresis. As the content of charcoal in
the soil increased, the value of H also increased. For the soils
Published on October 11, 2006 on http://pubs.acs.org | doi: 10.1021/jf061354y

amended with charcoal BC850, the H values increased rapidly

from 1.32 for the soil with 0.1% of charcoal to 14.92 for 1.0%
of charcoal. For the soils amended with charcoal BC450 the
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H value also progressively increased but at a slower rate. The

results showed that besides the high sorption capacity, charcoal
produced under higher temperature either had stronger sorption
or provided domains for diffusion of sorbate into micropores.
In this way the more microporous structure may lead to effective
sequestration of a compound. This is consistent with the studies
reporting poor bioavailability and bioaccessibility of organic
compounds sorbed on black carbon materials (40, 41).
Comparison of the results for the two types of charcoal
materials also revealed that although the sorption capacity of a
soil could be enhanced to the same level by adding differential
amounts of chars (0.8% BC850 vs 5% BC450), the degree of
reversibility was quite different between the two charcoals. In
the present study, the discrepancy between sorption and de-
sorption isotherms at comparable levels of sorption (e.g., 0.8%
BC850 vs 5% BC450, Table 2) was clearly much more
prominent in the case of charcoal BC850.
To assess if a link between the microporosity and apparent
desorption hysteresis exists, we plotted the values of the total
pore volume for each sorbent calculated from the V pore total value
and the content of charcoal material in the soil against the
apparent hysteresis index (H) in Figure 5. Here, we assumed
that the soil free of charcoal is a nonporous material and that
the pore volume of the charcoal-amended soils was mainly
provided by the added charcoal. Figure 5 shows a good rela-
tionship between these two parameters; the H index increased
exponentially (y ) 1.582 e0.008x, R2 ) 0.8982) as the total pore
volume of the sorbent increased. A smaller increase in H value
at lower pore volume is essentially associated with the presence
of charcoal BC450 and only small amounts of BC850. This is
because charcoal BC450 had a negligible proportion of micro-
pores, whereas in BC850 essentially all pores existed as micro-
pores of <1 nm width. These micropores either entrapoed the
sorbed molecules of diuron or caused a slow and prolonged
sorption phase, which may have led to the apparent hysteresis
due to nonequilibrium processes. Braida et al. (22) noted swell-
ing of a sorbent during benzene sorption and suggested pore
deformation during desorption causing desorption hysteresis.
It has also been suggested in other studies that surface-specific
adsorption, entrapment into micropores, and partitioning into
Figure 5. Relationship between total pore volume and apparent hysteresis
index (H).
condensed structures of soil organic matter are among the main
causes of chemical sequestration (35-37, 42-45).
Conclusions. This study showed that the presence of small
amounts of black carbon in the form of charcoals in soil,
especially those produced at high temperatures (e.g., interior
of wood logs during a fire), can have a major effect on the
sorption and desorption behaviors of organic compounds such
as the diuron pesticide. The marked effect on the desorption of
highly carbonaceous materials such as charcoal produced at high
temperatures is expected to have strong implications for the
bioavailability of such compounds in terrestrial and aquatic
ecosystems. The role of such carbonaceous materials on
sorption-desorption kinetics and bioavailability needs to be
further investigated.
ACKNOWLEDGMENT
We acknowledge the contributions by Drs. Phillip Pendleton
and Alexander Badalyan (University of South Australia) in
measuring pore size distribution; Jan Skjemstad and Evelyn
Krull (CSIRO) for advice and comments on synthetic char
production; Tasha Waller and Sonia Grocke (CSIRO) for
technical support; and Dr. Ron Smernik (University of Adelaide)
for helpful comments on the manuscript.
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Received for review May 12, 2006. Revised manuscript received August
31, 2006. Accepted September 4, 2006.
JF061354Y
 Science of the Total Environment 354 (2006) 170 – 178
www.elsevier.com/locate/scitotenv

Bioavailability of diuron in soil containing

wheat-straw-derived char
Yaning Yanga,c, Guangyao Shenga,T, Minsheng Huangb
a
Department of Crop, Soil, and Environmental Sciences, University of Arkansas, Fayetteville, AR 72701, U.S.A.
b
Department of Environmental Science and Technology, East China Normal University, Shanghai 200062, P.R. China
c
Department of Civil and Environmental Engineering, University of Illinois, Urbana, IL 61801, U.S.A.
Received 19 October 2004; accepted 26 January 2005
Available online 23 March 2005

Abstract

This study evaluated the bioavailability of diuron in soil as influenced by char arising from the burning of wheat straw. The wheat
char was a highly effective sorbent for diuron. The presence of 1% wheat char in soil resulted in a 7–80 times higher diuron sorption.
A 10-week incubation resulted in b40% of 0.5 mg/kg diuron in 0.5% char-amended soil microbially degraded, as compared to 50%
in char-free soil under the same conditions. Over the experimental range of diuron application rates from 0 to 12 mg/kg and of char
contents from 0% to 1.0%, a 4-week bioassay indicated that both the barnyardgrass survival rating and the fresh weight of
aboveground biomass decreased with increasing diuron application at given char contents but increased with increasing char content
at potentially damaging diuron application rates. Residual analyses of bioassayed soils showed that the soils with char contents of
0.5% and higher and diuron application rates of 3.0 mg/kg and higher, as compared to those with no or low (0.05%) char and a diuron
application rate of 1.5 mg/kg, had higher residual diuron levels but higher barnyardgrass survival ratings and fresh weights. These
results suggest that enhanced sorption of diuron in soil in the presence of wheat char reduced the bioavailability of diuron, as
manifested by reduced microbial degradation of diuron and its herbicidal efficacy to barnyardgrass. This study may have greater
implication than for burning of wheat straw that field burning of vegetations may reduce bioavailability of pesticides.
D 2005 Elsevier B.V. All rights reserved.

Keywords: Bioavailability; Diuron; Microorganism; Barnyardgrass; Sorption; Wheat char

1. Introduction degradation (Ogram et al., 1985; Shimp and Young,

Many studies have suggested that soil-bound
organic contaminants are unavailable for microbial
T Corresponding author. Tel.: +1 479 575 6752; fax: +1 479 575
3975.
E-mail address: gsheng@uark.edu (G. Sheng).
0048-9697/$ - see front matter D 2005 Elsevier B.V. All rights reserved.
doi:10.1016/j.scitotenv.2005.01.026
1988; Steen et al., 1980). Although recent research
suggested that limited biodegradation of soil-sorbed
pesticides may occur (Feng et al., 2000; Park et al.,
2001, 2002, 2003), liquid-phase contaminants are
much more bioaccessible to soil microorganisms
(Ogram et al., 1985; Guerin and Boyd, 1992; Lahlou
and Ortega-Calvo, 1999). Other studies have estab-
 Y. Yang et al. / Science of the Total Environment 354 (2006) 170–178
lished that phytotoxicity of herbicides is directly
related to their concentrations in the soil solution
(e.g., Lambert, 1966; Pillay and Tchan, 1971). It is
generally true that only dissolved nutrients and
organic chemicals in the soil interstitial bulk-like
water (i.e., plant-available water above the wilting
point) are available for plant-root uptake. Sorption
largely controls concentrations of organic contami-
nants in the soil solution and thus is the major
determinant of the bioavailability of pesticides to both
microorganisms and plants. Enhancing sorption by
increasing soil organic matter content leading to a
reduction in the solution-phase concentrations results
in a decreased microbial degradation of organic
contaminants (e.g., Guerin and Boyd, 1993). Charcoal
dipping or banding effectively protects many plants
from herbicidal injury (Arle et al., 1948; Burr et al.,
1972; Chandler et al., 1978; Jordan and Smith, 1971;
William and Romanowski, 1972).
Sorptive properties of agricultural soils, and thus
bioavailability of pesticides, are often influenced by
agricultural practices. Field burning of crop residues
worldwide incorporates the resulting chars into soils.
Hilton and Yuen (1963) postulated that the retained
sorptivity of many Hawaiian soils for ureas and s-
triazines after oxidative removal of soil organic matter
by hydrogen peroxide was a result of the peroxide-
resistant soil chars arising from burning of sugarcane
trash. Toth et al. (1999) observed a reduction in the
phytotoxicity of diuron applied over the ash of
recently burned kangaroo grass, due primarily to the
diuron sorption by the ash. We confirmed their
postulation by measuring the sorption of pesticides
on soil-free chars from burning of crop residues.
Sorption of diuron by the chars from burning of wheat
straw and rice residue was 400–2500 times higher
than that by a soil with 2.1% organic matter (Yang and
Sheng, 2003a). Amendment of the soil with the wheat
char up to 1% (by weight) enhanced the sorptivity of
the soil for diuron in proportion to the char content.
The char aged in the soil for one month remained
highly effective for diuron sorption and dominated the
sorption, although a small reduction (b30%) in
sorptivity was observed (Yang and Sheng, 2003b).
Further aging of the char for up to 12 months did not
result in a further sorptivity reduction, indicating that
the char was resistant to degradation. One direct
consequence of the high sorptivity of crop-residue-
171
derived chars may be the reduced bioavailability of
pesticides in soils to both microbes and plants. The
reduction in microbial degradation of benzonitrile in a
soil in the presence of wheat char has been reported
(Zhang et al., 2004). Reduced microbial degradation
increases the persistence of pesticides and thus the
environmental risk associated with pesticide use. The
high sorptivity of crop-residue-derived chars may also
reduce herbicidal efficacy to weeds. Poor herbicidal
efficacy is a concern both economically and environ-
mentally due to additional application of herbicides
for weed control.
In this study, we determined the bioavailability of
diuron to soil microorganisms and to barnyardgrass
(Echinochloa crus-galli (L.) Beauv.) in soil in the
presence of wheat-straw-derived char. Use of bar-
nyardgrass, a common rice weed, has both agronomic
and environmental significance. Our objectives were
to determine the microbial degradation of diuron and
its efficacy to barnyardgrass in relation to soil
sorption in the presence of wheat char and to evaluate
the impact of crop-residue-derived chars on the
bioavailability of pesticides in soil. Results from
laboratory measurements and greenhouse bioassays
indicated that wheat char was a highly effective
sorbent for diuron and its presence in soil resulted in
enhanced diuron sorption and subsequently dimin-
ished bioavailability.
2. Materials and methods
2.1. Materials
Wheat char used in this study was from our
previous studies (Yang and Sheng, 2003a,b). Air-
dried wheat (Triticum aestivum L.) straw (10 kg) was
collected from the Arkansas Agricultural Research
and Extension Center in Fayetteville, Arkansas.
Wheat char was obtained by burning the straw on a
stainless steel plate (1 m  1 m) in an open field under
natural conditions in a July afternoon. The BET
surface area of the char was determined to be 10.1 m2/
g in a commercial service laboratory. Chemical
analysis showed that the char contained 13% elemen-
tal carbon and 42% silica.
Soil, classified as Stuttgart silt loam, was collected
at the Rice Research and Extension Center, Stuttgart,
172 Y. Yang et al. / Science of the Total Environment 354 (2006) 170–178
Arkansas. The soil had a mechanical composition of
17.1% sand, 60.4% silt, and 22.5% clay. The soil
contained 2.1% organic matter and had a cation-
exchange capacity of 8.5 cmolc/kg. The soil, without
records of crop residue burns, was presumed to contain
minimal levels of chars. The soil was air-dried, ground,
and passed through a 1-mm sieve. Char-amended soils
were prepared by thoroughly mixing soil with accu-
rately weighed char in exact char contents of 0.05%,
0.1%, 0.5%, and 1.0% (by weight).
Diuron with a purity of 99% was purchased from
ChemService (West Chester, PA) and used as received.
Diuron is an electroneutral molecule. The pesticide has
a water solubility of ~42 mg/l and a log K ow of 2.68 at
room temperature (Howard and Meylan, 1997). The
Henry’s Law constant of 2.710 6 atm m3/mol
indicates that the pesticide is non-volatile and therefore
suitable for prolonged laboratory and greenhouse
studies. Diuron is a urea compound primarily used as
a preemergence herbicide in soils to control germinat-
ing grasses and broad-leaved weeds. As a photosyn-
thesis inhibitor, diuron injures weeds with symptoms
of foliar chlorosis concentrated around veins (some-
times interveinal) followed by necrosis.
A mixed diuron-degrading culture was isolated
from soil collected in a cotton field where diuron has
been applied annually for over 15 years. One gram of
the soil was inoculated into 100 ml of medium
containing mineral salts (Stanier et al., 1966), 0.1 ml
of vitamin solution (Wolin et al., 1963), and saturated
diuron. The mixture was incubated for 1 week at 28
8C on a platform shaker at 150 rpm. After two serial
transfers to fresh medium, the culture enrichment was
obtained. Preliminary tests showed that the isolated
culture readily degraded diuron in solution.
Barnyardgrass seeds, collected from the plant
grown in a rice field in Stuttgart, AR in 1983, were
acid-scarified in 1 N nitric acid for 1 h at room
temperature. The seeds were washed with deionized
water, spread evenly on filter paper in petri dishes,
covered with another filter paper, watered with 3 ml of
deionized water, and incubated in the dark at room
temperature for 72 h for germination.
2.2. Sorption isotherms for diuron
Sorption of diuron by soil, wheat char, and 1%
char-amended soil was measured by the batch
equilibration technique. Various quantities of diuron
in 0.005 M CaCl2 solution were added into 25-ml
Corex glass centrifuge tubes containing sorbents with
a constant mass between 0.01 and 3.5 g. The mass of
sorbents was adjusted to allow for N40% of added
diuron to be adsorbed. Additional 0.005 M CaCl2
solution was added to bring the total liquid volume to
10 ml. Initial concentrations of diuron ranged from
1.25 to 12.5 mg/l. The tubes were closed with Teflon-
lined screw caps and rotated (40 rpm) at room
temperature (~25 8C) for 48 h. Other measurements
have shown that sorption of diuron by both the soil
and the char reached apparent equilibria within 24 h;
the sorption by char-amended soil was thus assumed
to also reach equilibrium within the same duration.
After the establishment of sorption equilibria,
sorbents and aqueous phases were separated by
centrifugation at 6000 rpm (RCF=5210 g) for 30
min. The diuron concentrations in supernatants were
analyzed by high-performance liquid chromatography
(HPLC). The amount of diuron sorbed was calculated
from the difference between the amount initially
added and that remaining in equilibrium solution.
All measurements were in duplicate with a variation
generally b5%, and the calculated average data were
reported. The measurements with blanks not contain-
ing sorbents found that glass tubes did not adsorb
diuron and no processes other than sorption con-
tributed to the loss of solution-phase diuron.
2.3. Microbial degradation of diuron
Sterilized soil and 0.5% char-amended soil (200 g
each) were placed in 1000-ml glass beakers, spiked
with 2 ml of 50 mg/l diuron stock solution in acetone,
thoroughly mixed, and allowed for acetone to
evaporate for 2 days. The soils were then watered
to near the field capacity and aged for another 2 days
to allow the diuron sorption to complete. Three 5-g
samples from each of the soils were extracted with 5
ml of H2O and 5 ml of water-saturated ethyl acetate
for 72 h. Following the phase separation, 2 ml of
ethyl acetate were dried under N2 gas and redissolved
in 1 ml of methanol. The diuron concentrations in
methanol were analyzed by HPLC. The recoveries
were 95.2% and 85.2% for soil and char-amended
soil, respectively. Each soil was inoculated with 5 ml
of the isolated culture enrichment. Preliminary tests
 Y. Yang et al. / Science of the Total Environment 354 (2006) 170–178
found that 5 ml of the enrichment in 200 g of soil
gave an appreciable degree of diuron degradation in
one week. The soils were thoroughly mixed again.
The beakers were then covered with aluminum foil
and placed in the dark at room temperature. Three 5-g
soil samples from each beaker were taken weekly for
10 weeks for diuron analysis. The soils were
extracted following the same extraction procedures
as those for the recoveries, except that extracting
water contained 0.5% Ag2SO4 to immediately termi-
nate biodegradation. The diuron concentrations were
adjusted for the recoveries. The soils, watered when
necessary, were maintained moist throughout the
degradation experiment.
2.4. Barnyardgrass bioassay for diuron
A series of 4 diuron solutions with concentrations
of 60, 120, 240, and 480 mg/l in acetone were
prepared. Five milliliters of each of the solutions,
along with acetone only, were added to 200 g of char-
free soil or char-amended soils in plastic bags,
resulting in the diuron levels of 0, 1.5, 3.0, 6.0, and
12 mg/kg in the soils, respectively. The five diuron
levels were referred to as 0X, 1X, 2X, 4X, and 8X,
where X=1.5 mg/kg and is within the range of
recommended field application rates. The five char
contents were 0%, 0.05%, 0.1%, 0.5%, and 1%. The
combination of the 5 diuron rates and the 5 char
contents resulted in a total of 25 treatments. Each
treatment was in triplicate. The soils were thoroughly
mixed in the bags, and transferred to plastic cups
following the evaporation of acetone from the opened
bags.
A small quantity of soil (~20 g) was first removed
from each cup. Ten pregerminated barnyardgrass
seeds with extended radicles and hypocotyls were
placed evenly on the soil surface in each cup and then
covered with the previously removed soil. Following
planting, the cups were placed in a completely
randomized block design in a greenhouse, watered
with 40 ml of deionized water, and maintained moist
throughout the experiment. The greenhouse was
maintained at 34/20 8C day/night temperatures with
a 14-h lighting cycle. Barnyardgrass seedling survival
was visually rated two weeks after planting as percent
of survival between 0 and 100, with 0% representing
no survival and 100% complete survival (no injury).
173
The survival rating was performed independently by
three individuals. Average survival ratings were
calculated using the data from the replicated samples
from all the individuals. Four weeks after planting,
plants were cut at the soil level and immediately
weighed to obtain the fresh weights of the above-
ground biomass. All the visual survival rating and
fresh weight data were statistically analyzed using the
SAS program.
Following the bioassay, the soil samples of selected
treatments were analyzed for residual diuron by
HPLC using the extraction procedures described
earlier. The average diuron concentration of the
replicate soils of the same treatment was reported.
The soils from the following 6 treatments were
selected based on the char contents, diuron rates,
and barnyardgrass aboveground fresh weights: S0-0,
S0-1.5, S0.05-1.5, S0.5-3.0, S1-3.0, and S1-6.0,
where the suffix C-D to S represents the char content
(C)-applied soil concentration of diuron (D).
2.5. Analysis of diuron
Diuron in the supernatants from sorption experi-
ments and in the extracts from microbial degradation
and greenhouse bioassay tests was analyzed on a
Hitachi reversed-phase high-performance liquid chro-
matograph (Hitachi High-Technologies Tokyo,
Japan) fitted with a UV-visible detector set at the
maximum absorption wavelength for diuron (252
nm). A Phenomenex Prodigy C18 column was used
(Alltech Assoc., Deerfield, IL). The mobile phase
was a mixture of acetonitrile and water (50:50, v/v)
at a flow rate of 1.0 ml/min. The injection volume
was 20 Al.
3. Results and discussion
Isotherms for the sorption of diuron by soil, wheat
char and 1% char-amended soil are presented in Fig.
1, in which the amount of diuron sorbed (mg/kg) is
plotted against the equilibrium concentration (mg/l) in
water. No single mathematical models provided
adequate fits for the sorption data. The curves were
drawn to assist in visualization and comparison of the
data. Soil effectively sorbed diuron, rather consistent
with the prediction from the log K ow value of diuron
174
100
Amount of Diuron Sorbed (mg/kg)
Soil
1% Wheat Char in Soil
80
60
40
20
0 simply measured the dissipation of diuron over
0 2
Equilibrium Concentration (mg/L)
Fig. 1. Isotherms for sorption of diuron from water by soil, wheat
char, and char-amended soil.
and the soil organic matter content. Wheat char had a
much higher sorptivity than soil for diuron (see the
inset in Fig. 1). From the curves, we estimated that the
char was 700–37,000 times more effective than the
soil in sorbing diuron over the experimental concen-
tration range (0–6 mg/l). These results are similar to
those obtained in our previous study (Yang and
Sheng, 2003a). It has also been reported in the
literature that chars (the term ash was used) derived
from burning of vegetation are effective adsorbents
for pesticides (Yang and Sheng, 2003b; Toth and
Milham, 1975).
Much higher sorptivity of wheat char than of soil
resulted in enhanced sorption of diuron by the soil in
the presence of the char, as indicated in Fig. 1.
Calculations show that 1% char-amended soil sorbed
about 7–80 times more diuron than char-free soil over
the experimental concentration range. Assuming that
the amendment of soil with 1% char did not change
the sorptivity of the soil for diuron, 1% char
contributed N86% to the total sorption of diuron,
indicating the predominance of the char for diuron
sorption. Normalization of the sorption to char content
resulted in a diuron isotherm slightly lower than that
for soil-free char, which may have resulted from the
competitive sorption of dissolved soil organic matter
on the char. While the diuron sorption by char-
amended soil was evaluated with only one char
content, enhanced sorption at other char contents is
expected. At recommended field application rates,
enhanced diuron sorption in the presence of wheat
Y. Yang et al. / Science of the Total Environment 354 (2006) 170–178
char may decrease the concentration of diuron in the
soil solution, leading to reduced biodegradation and
loss of herbicidal efficacy.
An evaluation of the bioavailability of diuron to
9000
soil microorganisms in the presence and absence of
6000
wheat char in soil was made by comparing the
3000 Wheat Char
degradation of diuron in char-free soil and 0.5%
0
0 2 4 6 char-amended soil, both inoculated with the same size
of an isolated diuron-degrading culture. The initial
diuron concentration in both soils was 0.5 mg/kg. We
4 6 8 10 incubation time and did not identify the degradation
products, thus offering no mechanistical information
on the diuron transformation reaction. In Fig. 2, the
degradation is expressed as the percent of total diuron
degraded at given time. Although diuron is a rather
persistent pesticide in field soils, it slowly degraded in
field soils (Hill et al., 1955). It has been reported that
mixed cultures from pond water and sediment aerobi-
cally degraded diuron to several identified products
and carbon dioxide (Ellis and Camper, 1982). While
we did not know the species and the population of the
isolated cultures, diuron in both soils was degraded
with time. The degradation was slower in char-
amended soil than in char-free soil. Over a 10-week
incubation, b40% of diuron in 0.5% char-amended
soil was degraded, in comparison to about 55% in
char-free soil. While direct degradation of soil-sorbed
organic compounds by bacteria may occur, we did not
know whether such a process was involved in diuron
degradation. However, our results clearly show that
60
50
Diuron Degraded (%)
40
30
20
Soil
10 0.5% Wheat Char in Soil
0
0 20 40 60 80
Incubation Time (days)
Fig. 2. Microbial degradation of diuron over time in sterilized soil
and char-amended soil inoculated with a mixed enrichment culture.
 Y. Yang et al. / Science of the Total Environment 354 (2006) 170–178
enhanced sorption in the presence of wheat char
reduced the bioavailability of diuron to soil micro-
organisms. Reduced biodegradation of benzonitrile by
a bacterium in soil in the presence of wheat char was
also due to enhanced sorption (Zhang et al., 2004). In
addition to elemental carbon and silica, wheat char
contained 21% potassium, 1.5% phosphorous, 0.64%
nitrogen, and other microelements (Yang and Sheng,
2003b). When in their available forms in soil, some of
these nutrients may stimulate microbial activity. We
have found that when benzonitrile was not limiting,
1% wheat char provided nutritional stimulation on
benzonitrile degradation (data not shown). Such a
stimulation on diuron degradation in 0.5% char-
amended soil was not obvious.
Fig. 3 is the photograph comparing the barnyard-
grass growth among the soil samples subjected to
diuron applications in the absence and presence of
wheat char just prior to cutting the plants for their
fresh weights. The samples consisted of 25 cups, each
representing one of the three replicates for each of all
the treatments, and were placed de-randomized to aid
visualization. In the absence of diuron application,
barnyardgrass showed a normal growth without
observable growth stimulation by wheat char
nutrients. One week after planting, the barnyardgrass
injury was obvious (photograph not shown). The
injury increased with increasing diuron application
0.0 1.5
Application Rate of Diuron (mg/kg)
Fig. 3. Photograph showing barnyardgrass growth in soils as a function of diuron application rate and wheat char content four weeks after
planting.
175
rate at a given char content and decreased with
increasing char content at a given diuron application
rate. The observation is consistent with the earlier
sorption measurements that the presence of wheat
char enhanced the diuron sorption by soil. By four
weeks after planting (prior to cutting), uninjured
barnyardgrass showed continued growth, whereas
injured ones did not recover (Fig. 3).
More quantitative barnyardgrass bioassay to eval-
uate the impact of wheat char on the bioavailability of
diuron was obtained by visually rating barnyardgrass
survival two weeks after planting and by weighing
aboveground fresh biomass four weeks after planting
(Fig. 4). Without application of diuron, barnyardgrass
was somehow injured in char-free soil. In the presence
of wheat char (z0.05%), the injury was eliminated.
This suggests that unknown herbicides likely phyto-
toxic to barnyardgrass may be present in char-free soil
but deactivated by the char. A full rate of diuron (1.5
mg/kg) in char-free soil showed almost complete
injury to barnyardgrass. However, both the barnyard-
grass survival rating and fresh weight increased with
increasing char content. A char content of 0.5% or
higher was sufficiently high that diuron completely
lost its efficacy to barnyardgrass. The application rates
of 3 mg/kg and higher resulted in complete-to-partial
losses of diuron efficacy to barnyardgrass. Similar to
the observations with the full rate of diuron, the
Wheat Char in Soil (%)
1.0
0.5
0.1
0.05
0.0
3.0 6.0 12
176 Y. Yang et al. / Science of the Total Environment 354 (2006) 170–178

(a) (b)
100 0.05
(%)

0.04

)
80

Fresh Weight (g
Survival Rating

60 0.03

40 0.02

20 1.0 0.01 1.0

)
0.5 0.5

(%

(%
0 0.1 0.00 0.1

nt

nt
0 0.05 0 0.05
nte

n te
1.5 3.0 0 Co 1.5 3.0 0

Co
Rate 6.0 Rate 6.0
of Diu 12 of Diu 12
ron (m
ar

ar
ron (m
g/kg)
Ch

Ch
g/kg)

Fig. 4. Growth of barnyardgrass in soils as a function of diuron application rate and wheat char content showing (a) survival rating of
barnyardgrass two weeks after planting, and (b) fresh weight of barnyardgrass four weeks after planting.

survival rating and fresh weight at higher application contained residual diuron with levels of about 40–
rates increased with increasing char content over the 49% of their respective application rates. The soils S0-
tested range of char contents in soil. The Student’s t 1.5 and S0.05-1.5 containing no or low char (0% and
test at the 95% level of confidence (a=0.05) was used 0.05%, respectively) had residual diuron concentra-
to compare the individual survival ratings and fresh tions of ~0.7 mg/kg and produced much lower
biomass weights at various char contents and diuron barnyardgrass fresh weights than the soil S0-0,
application rates. Survival ratings and fresh weights indicating the availability of diuron to barnyardgrass
were statistically different (a=0.05) for V0.05% char in these soils. Although the soils S0.5-3.0 and S1-3.0
with no diuron application, for V0.5% char with 1.5 containing 0.5% and 1% char, respectively, had
mg diuron/kg, for z0.1% char with 3.0 mg or 6.0 mg residual diuron concentrations almost twice those in
diuron/kg (fresh weights were significantly different the soils S0-1.5 and S0.05-1.5, the barnyardgrass
forz0.5% char with 6.0 mg diuron/kg), and for fresh weights associated with the former two soils
z0.5% char with 12 mg diuron/kg. These results were much higher than those with the latter two. In
suggest the decreased bioavailability of diuron to fact, the barnyardgrass fresh weights with the soils
barnyardgrass with increasing char content in soil, due
presumably to enhanced sorption of diuron in the Table 1
presence of the char. Relationship between measured residual concentrations of diuron
The sorptive role of wheat char in reducing diuron and barnyardgrass fresh weights in selected soil and char-amended
bioavailability (efficacy) to barnyardgrass is con- soil samples subjected to various treatments four weeks after
firmed by measuring residual concentrations of diuron planting as influenced by percent char content and application rate
of diuron
in soils after cutting barnyardgrass. Soils from 6
selected treatments, where barnyardgrass fresh Soil Char Rate of Barnyardgrass Residual
content diuron fresh fresh weight (g) concentration
weights differed significantly, were analyzed for (%) weight (g) (mg/kg)
residual diuron. The concentrations in the three
S0-0 0 0 0.0242 0.00
replicate soils of each treatment were highly invariant, S0-1.5 0 1.5 0.0007 0.73
with the difference b4.3%. The average concentra- S0.05-1.5 0.05 1.5 0.0030 0.69
tions were calculated and presented in Table 1. Char- S0.5-3.0 0.5 3.0 0.0224 1.28
free soil (S0-0) did not contain a measurable level of S1-3.0 1 3.0 0.0308 1.20
S1-6.0 1 6.0 0.0378 2.42
diuron. All other soils that had received diuron
 Y. Yang et al. / Science of the Total Environment 354 (2006) 170–178
S0.5-3.0 and S1-3.0 were similar to that with char-free
soil without receiving diuron application (i.e. the soil
S0-0). The barnyardgrass fresh weight with the soil
S1-6.0 containing an even higher level of residual
diuron remained high. These results indicate that
diuron in soils with high contents of wheat char was
largely unavailable to barnyardgrass.
Our measurements indicate that at typical applica-
tion rates of diuron, a char content of as low as 0.1%
may appreciably reduce the bioavailability of diuron
in soil. We reported that burning of wheat straw
produced wheat char at ~6% of the straw weight
(Yang and Sheng, 2003a). Using the average produc-
tion of wheat straw of ca. 6000 kg/ha, its burning
would generate ~360 kg/ha wheat char. If this wheat
char were mixed in soil with a density of 1.4 g/cm3 to
the depth of furrow slice (~0.15 m), a single burning
would result in a wheat char content of~0.02%. Crop-
residue-derived chars are expected to accumulate in
soils, as crop residues are repeatedly burned, and the
resulting chars are expected to remain to be highly
effective sorbents for pesticides (Yang and Sheng,
2003b). As such, field burning of crop residues may
effectively reduce the bioavailability of pesticides.
4. Conclusions
Wheat char is a highly effective sorbent for diuron.
Field burning of wheat straw incorporates the resulting
wheat char into soil and enhances the sorption of
diuron by the soil. A direct consequence of this
agricultural practice is the reduced bioavailability of
diuron in soil. We found that diuron was less
biodegradable in soil in the presence of wheat char.
Its herbicidal efficacy to barnyardgrass decreased with
increasing char content in soil and, at recommended
field application rates, could be completely lost when
the soil char content was 0.5% or higher. Reduced
bioavailability of diuron appeared to result from the
enhanced sorption in soil in the presence of wheat char.
Although only wheat char and diuron were tested in
this study, it is expected that chars arising from field
burning of other crop residues and vegetations also
effectively sorb other pesticides and reduce their
bioavailability. The presence of crop-residue-derived
chars in soil may increase the environmental risk of
pesticides and reduce their efficacy to pests.
177
Acknowledgments
This research was supported by USDA National
Research Initiative Competitive Grants Program
(Grant No. 2002-35107-12350).
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Note
Simultaneous biodegradation of chloro-
and methylthio-s-triazines using
charcoal enriched with a newly
developed bacterial consortium
Ken-ichi YAMAZAKI,† Kazuhiro TAKAGI*,†,††
Kunihiko FUJII,††† Akio IWASAKI,†††
Naoki HARADA†††† and Tai UCHIMURA†
†
Department of Applied Biology and Chemistry, Tokyo University of
Agriculture, Setagaya-ku, Tokyo 156–8502, Japan
††
Organochemicals Division, National Institute for
Agro-Environmental Sciences, Tsukuba, Ibaraki 305–8604, Japan
†††
Kowa Research Institute, Kowa Co., Ltd.,
Tsukuba, Ibaraki 305–0856, Japan
††††
Faculty of Agriculture, Niigata University,
Nishi-ku, Niigata 950–2181, Japan
(Received February 18, 2008; Accepted April 10, 2008)
A special type of charcoal, Charcoal A100, was enriched with a
newly developed bacterial consortium using a perfusion method.
The bacterial consortium consisted of a methylthio-s-triazine-de-
grading bacterium (Rhodococcus sp. FJ1117YT) and the chloro-
s-triazine-degrading bacterial consortium CD7 (containing
Bradyrhizobium japonicam CSB1, Arthrobacter sp. CD7w and
b -Proteobacteria CDB21). Enriched charcoal was capable
of degrading chloro-s-triazines (simazine and atrazine) and
methylthio-s-triazines (simetryn and dimethametryn) simultane-
ously in sulfur-free medium. Almost complete degradation was
observed after 4-day cultivation of chloro-s-triazines and 9-day
cultivation of methylthio-s-triazines. These triazines were miner-
alized via their 2-hydroxy analogues.
Keywords: biodegradation, methylthio-s-triazines, chloro-s-tri-
azines, bacterial consortium, Charcoal A100.
Introduction
The s-triazines are recognized as a major class of herbicides and
are widely used in agriculture for controlling various weeds, and
they have been detected in ground and surface water.1–3) Among
the removal technologies for such residual pesticides, bioremedi-
ation is considered to be the most cost-effective and safe technol-
ogy; therefore, many studies concerning s-triazine-degrading
* To whom correspondence should be addressed.
E-mail: ktakagi@niaes.affrc.go.jp
Published online July 14, 2008
© Pesticide Science Society of Japan
J. Pestic. Sci., 33(3), 266–270 (2008)
DOI: 10.1584/jpestics.G08-08
bacteria have been reported.
In our study, bacterial consortium CD74) was obtained which
can mineralize simazine, and a degrading bacterium b -Pro-
teobacteria CDB21 was isolated from CD7 by Iwasaki et al.5) It
was considered that CD7 was more effective for bioremediation
than strain CDB21 because strain CDB21 could not utilize
simazine for its growth in mineral salt medium without
cyanocobalamin, but CD7 could utilize simazine as sole carbon
and nitrogen sources in mineral salt medium.4,5) In addition, a
special type of charcoal, Charcoal A100, enriched with CD7 was
developed, and was placed under the subsoil of a golf course to
degrade simazine. The simazine-degrading capability of the en-
riched charcoal was maintained for nearly 2 years.4) Thus, Char-
coal A100 enriched with CD7 is considered to be an effective
material for bioremediation; however, CD7 and strain CDB21
could not degrade methylthio-s-triazines detected in river
water,3,6) a lake basin,7) and river sediments,8) while Rhodococcus
sp. FJ1117YT9) can transform methylthio-s-triazines to their hy-
droxyl analogues via sulfur oxidation, and accumulate hydroxy-s-
triazines. It is considered that simultaneous degradation of
chloro- and methylthio-s-triazines using a mixture of these bacte-
ria is meaningful because river water3,10) and river sediment3)
contamination with both triazines has been reported.
To take advantage of these useful properties of CD7 and Char-
coal A100, this study describes the development of Charcoal
A100 enriched with strain FJ1117YT and CD7, and demonstra-
tion of the simultaneous degradation of chloro- and methylthio-s-
triazines using this novel material.
Materials and Methods
1. Materials
Simazine, atrazine, simetryn, and dimethametryn were purchased
from Kanto Kagaku. Cyanuric acid and simazine-2-hydroxy were
purchased from Dr. Ehrenstorfer GmbH. The sulfur-free mineral
salt medium (MM) used in this study contained Na2HPO4 · 12H2O
1.2 g/l and KH2PO4 0.5 g/l. Prior to autoclaving, the medium was
supplemented with 10 ml/l of a sulfur-free solution of trace ele-
ments, which contained MgCl2 · 6H2O 2000 mg/l, FeCl2 · H2O 200
mg/l, ZnCl2 10 mg/l, MnCl2 5 mg/l, CoCl2 24 mg/l, CuCl2 5 mg/l,
NiCl2 · 6H2O 5 mg/l, Na2MoO4 5 mg/l, H3BO4 30 mg/l, Ca(OH)2
50 mg/l, and EDTA 500 mg/l. Mineral salt medium containing
sulfate (MM
S) was also used. MM
S contained MgSO4 20
mg/l and a solution of trace elements described by Yanze-
Kontchou and Gschwind.11) MM and MM
S were supplemented
with s-triazines before autoclaving. Vitamin mixture was used as
described by Fujii et al.9)
2. Analytical methods
The concentration of s-triazines in the medium was determined
using HPLC (Tosoh) using the following method. Separation of
s-triazines was achieved at 40°C on an ODS column (Capcell Pak
C18 UG120 3250 mm, 5 m m particle size; Shiseido) with a mo-
Vol. 33, No. 3, 266–270 (2008)
bile phase containing a gradient mixture of acetonitrile and water
(20%/80% 0 min–20%/80% 5 min, linear gradient–40%/60% 11
min, and linear gradient–20%/80% 15 min–20%/80% 20 min).
The injection volume was 50 m l and the eluted compounds were
detected by UV at 220 nm. Metabolites of s-triazines were deter-
mined using LC-ESI-MS. Separation was achieved using LC Al-
liance 2695 (Waters) system equipped with the same ODS col-
umn as above (40°C). A mixture of acetonitrile/0.2% acetic acid
was used as the eluent (5%/95% 0 min, linear gradient–50%/50%
10 min–80%/20%10.1 min–80%/20% 15 min, and linear gradi-
ent–5%/95% 20 min). The injection volume was 10 m l. The ESI-
MS system was Quattro micro API (Waters) with the flow rate of
nebulization set at 100 l/h and the flow rate for desolvation gas
(N2) set at 500 l/h. The respective temperatures of source and
desolvation were set at 100 and 350°C; and capillary voltage was
3.5 kV. Detection was performed by scanning between m/z 100
and m/z 300 under cone voltage of 35 V in positive ion mode.
Cyanuric acid was detected at m/z 128 [MH] under cone volt-
age of 23 V in negative ion mode, and simazine-2-hydroxy was
m/z 184 [M
H]
under cone voltage of 35 V in positive ion
mode.
3. Enrichment of Charcoal A100 with strain FJ1117YT and
CD7
Charcoal A100 (Toyo Denka Kogyo) was used as the enrichment
material. Enrichment of charcoal with a bacterial consortium was
performed using a charcoal perfusion method, which was modi-
fied from the soil perfusion method described by Audus.12) First,
washed Charcoal A100 (7.5 g, dry weight) was packed in perfu-
sion apparatus equipped with glass sintered filter, and then auto-
claved. Subsequently, three pieces of stab cultures of CD7 on a
MM agar plate containing 40 mg/l simazine were placed on the
charcoal, and the charcoal layer was covered with glass fiber filter
paper. Enrichment was performed in the dark at 25°C. MM (300
ml) containing 5 mg/l simazine was perfused with air lift using an
air pump for 14 days. The concentration of simazine in the perfu-
sion fluid was determined by HPLC, and the perfusion fluid was
replaced twice. After 14 days, the glass fiber filter paper was re-
placed. A phosphate buffer suspension of strain FJ1117YT was
placed on the filter paper. MM (300 ml) containing 5 mg/l
simazine, 5 mg/l simetryn was perfused under similar conditions
for 21 days, and the perfusion fluid was replaced once during the
perfusion process. Lastly, MM (300 ml) containing 5 mg/l sime-
tryn was used as perfusion fluid for 38 days, and was replaced
once. The vitamin mixture was added twice. Total enrichment
time was 73 days, and this material was used thereafter. Non-en-
riched Charcoal A100 was also perfused as a control under the
same conditions.
4. Preparation of DNA and analysis of 16S rRNA gene
Adherent bacteria on the surface of Charcoal A100 enriched with
strain FJ1117YT and CD7 were removed by sonication in phos-
phate buffer for 2 min. Charcoal A100 inhabited with strain
FJ1117YT and CD7 was pulverized, and total DNA was ex-
tracted with Fast DNA Kit for soil (Q-Bio gene). Total DNA of
Simultaneous biodegradation of chloro- and methylthio-s-triazines 267
strain FJ1117YT and each member of CD7 were extracted from
suspensions of bacterial colonies grown on R2A agar (Difco)
plates by a protocol for gram-positive bacteria of DNeasy Blood
& Tissue Kit (QIAGEN). Nucleotide sequences of 16S rRNA
genes of isolated bacteria were analyzed by direct sequencing of
the PCR products described previously.5)
5. Denaturing gradient gel electrophoresis (DGGE) analy-
sis
Touchdown PCR was performed with a TP600 thermal cycler
(Takara BIO) using GoTaq Green Master Mix (Promega). GM5F-
gc-clamp and R534,13) primers designed from a sequence of the
variable V3 region of 16S rRNA, were used for PCR. DGGE
analysis was performed using 6% polyacrylamide gel (ratio of
acrylamide to bisacrylamide 37.5 : 1) in 1TAE (40 mM Tris
base, 20 mM acetic acid, and 1 mM EDTA) with a 30% to 60%
denaturant gradient (100% denaturant containing 7 M urea and
40% formamide). Electrophoresis was performed at a constant
voltage of 150 V and at a temperature of 60°C for 3.5 h by using
the DcodeTM universal mutation detection system (Bio-Rad).
After electrophoresis was completed, the gel was stained with
SYBR gold (Invitrogen) for 20 min, rinsed, and the bands were
visualized with a Molecular Imager Pharos FX plus system (Bio-
Rad).
6. Simultaneous degradation of chloro- and methylthio-s-tri-
azines using Charcoal A100 enriched with strain
FJ1117YT and CD7
Charcoal A100 enriched with strain FJ1117YT and CD7 (0.4 g
dry weight) was inoculated in MM (30 ml) containing 5 mg/l
each of simazine, atrazine, simetryn, and dimethametryn or in
MM
S containing 5 mg/l of each herbicide in 50 ml flasks. The
flasks were shaken at 120 rpm at 25°C for 15 days. As controls,
sterile Charcoal A100 and/or Charcoal A100 enriched with only
CD7 were inoculated and shaken under the same conditions. The
concentration of s-triazines in the medium was determined peri-
odically by HPLC. Cyanuric acid, simazine-2-hydroxy, and hy-
droxy analogues of atrazine and dimethametryn in the medium
after 15 days were measured by LC-ESI-MS.
Results and Discussion
1. Enrichment of Charcoal A100 with a bacterial consor-
tium
The extent of enrichment of Charcoal A100 with strain
FJ1117YT and CD7 was determined from the change in the con-
centration of simazine and simetryn in the perfusion fluid. In the
first step of enrichment, CD7 was enriched in Charcoal A100.
Concentration of simazine in the CD7-inoculated charcoal de-
creased faster than the control, and the degradation rate of
simazine increased with every replacement of the perfusion fluid
(data not shown). The charcoal was subsequently further enriched
with strain FJ1117YT and perfused with MM containing
simazine and simetryn. Simazine was degraded almost immedi-
ately (Fig. 1-A until Day21), in contrast to the slow degradation
of simetryn (Fig. 1-B until Day21). The disappearance rate of
 268 K.-i. Yamazaki et al. Journal of Pesticide Science

Fig. 1. Enrichment of strain FJ1117YT and CD7 in Charcoal A100 using a charcoal perfusion method. Changes in the concentrations of
simazine (A) and simetryn (B) during enrichment are illustrated (both figures overlapped until 21 days). Charcoal A100 without enrichment was
used as a control. In order to enrich CD7, MM containing 5 mg/l simazine was perfused for 14 days before these data. The arrows indicate as fol-
lows: replacement with MM containing simazine and simetryn (black), replacement with MM containing simetryn (white), and addition of vita-
min mixture (striped).

simetryn was initialing almost the same as that of control, but in-
creased after the second perfusion (Fig. 1-B on Day8). The de-
crease of pesticides in sterile Charcoal A100 was considered to
occur because pesticides were adsorbed on Charcoal A100. In
order to aid the growth of strain FJ1117YT,9) vitamin mixture
was added but had little effect on the enrichment of FJ1117YT.
However, because the enrichment of CD7 was sufficient, MM
containing simetryn without simazine was further perfused (Fig.
1-B after Day21), and the degradation rate of simetryn was fur-
ther increased. This result indicates the enrichment of strain
FJ1117YT in Charcoal A100.
less than 0.1 mg/l and hydroxy analogues of atrazine and
dimethametryn were not detected (data not shown). This result
suggests that hydroxyl analogues of methylthio-s-triazines, which
can not be metabolized by strain FJ1117YT, were mineralized by
CD7 via N-ammelide analogue and cyanuric acid. These results
also indicate that methylthio-s-triazines could be mineralized by
the bacterial consortium that included strain FJ1117YT and CD7.
In conclusion, the respective degradation abilities of CD7 and
strain FJ1117YT were successfully maintained in Charcoal

2. Detection of bacterial community in Charcoal A100 en-

riched with strain FJ1117YT and CD7
Colony isolation from the bacterial consortium CD7 and subse-
quent analyses of 16S rRNA genes revealed that CD7 consisted
of Bradyrhizobium japonicam CSB1, Arthrobacter sp. CD7w and
strain CDB21. The presence of all members of CD7 and strain
FJ1117YT enriched in Charcoal A100 were confirmed by PCR-
DGGE analyses (Fig. 2).

3. Simultaneous degradation of chloro- and methylthio-s-tri-

azines, and detection of their metabolites
Charcoal A100 enriched with strain FJ1117YT and CD7 was ap-
plied to simultaneous degradation of chloro- and methylthio-s-tri-
azines. Simazine and atrazine were degraded to by 80–100% with
Charcoal A100 enriched with both strain FJ1117YT and CD7 or
CD7 alone after 9 days, and were completely degraded after 15
days (Fig. 3-A, B). On the other hand, simetryn and dimethame-
tryn were degraded by over 80% with strain FJ1117YT and CD7 Fig. 2. PCR-DGGE band profiles of bacterial strains inhabiting
enriched charcoal in sulfur-free medium, but they were not de- Charcoal A100. Each lane represents DNA samples extracted from
graded in the presence of sulfate (in MM
S) (Fig 3-C, D). In the the following specimens: Charcoal A-100 enriched with FJ1117YT
sulfur-free medium with strain FJ1117YT and CD7 after 15 days, and CD7 (A); FJ1117YT (B); CDB21 (C); CSB1 (D); and CD7w
the concentrations of simazine-2-hydroxy and cyanuric acid were (E).
Vol. 33, No. 3, 266–270 (2008) Simultaneous biodegradation of chloro- and methylthio-s-triazines 269

Fig. 3. Time course of simultaneous degradation of chloro- and methylthio-s-triazines with Charcoal A100 enriched with strain FJ1117YT
and CD7. Degradation of simazine (A), atrazine (B), simetryn (C), and dimethametryn (D) by strain FJ1117YT and CD7 with (
) or without
() sulfate, with CD7 alone (), and using non-enriched Charcoal A100 as a control () are shown.

A100. Chloro- and methylthio-s-triazines were degraded simulta-
neously and their metabolites were hardly detected. The expected
metabolic pathways of chloro- and methylthio-s-triazines in the
mixed culture are shown in Fig. 4. Degradation of methylthio-s-
triazines by the bacterial consortium, strain FJ1117YT and CD7,
was suppressed by the presence of sulfate (Fig. 3) as well as the
culture of strain FJ1117YT reported previously.9) However, Char-
coal A100 enriched with strain FJ1117YT and CD7 could be a
promising model to construct a multifunctional material enriched
with bacterial consortium for in situ bioremediation. On the basis
of this study, we will attempt to construct another charcoal mate-
rial, which will include methylthio-s-triazines-degrading bacteria
that are not suppressed by external sulfur sources.

Fig. 4. The expected metabolic pathways of chloro- and methylthio-s-triazines degraded by CD7 (strain CDB21) and strain FJ1117YT.
Simazine (R1, R2C2H5) and atrazine [R1C2H5, R2CH(CH3)2] were selected as chloro-s-triazines, and simetryn (R1, R2C2H5) and
dimethametryn [R1C2H5, R2CH(CH3)CH(CH3)2] were used as methylthio-s-triazines in this study.
 270 K.-i. Yamazaki et al. Journal of Pesticide Science

Acknowledgments 5) A. Iwasaki, K. Takagi, Y. Yoshioka, K. Fujii, Y. Kojima and N.

This work was supported in part by a Grant-in-aid (Hazardous Chem-
icals) from the Ministry of Agriculture, Forestry, and Fisheries of
Japan (HC-05-2441-1).
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 EFFECT OF LOW‐TEMPERATURE PYROLYSIS CONDITIONS
ON BIOCHAR FOR AGRICULTURAL USE

J. W. Gaskin, C. Steiner, K. Harris, K. C. Das, B. Bibens

ABSTRACT. The removal of crop residues for bio‐energy production reduces the formation of soil organic carbon (SOC) and
therefore can have negative impacts on soil fertility. Pyrolysis (thermoconversion of biomass under anaerobic conditions)
generates liquid or gaseous fuels and a char (biochar) recalcitrant against decomposition. Biochar can be used to increase
SOC and cycle nutrients back into agricultural fields. In this case, crop residues can be used as a potential energy source as
well as to sequester carbon (C) and improve soil quality. To evaluate the agronomic potential of biochar, we analyzed biochar
produced from poultry litter, peanut hulls, and pine chips produced at 400°C and 500°C with or without steam activation.
The C content of the biochar ranged from 40% in the poultry litter (PL) biochar to 78% in the pine chip (PC) biochar. The
total and Mehlich I extractable nutrient concentrations in the biochar were strongly influenced by feedstock. Feedstock
nutrients (P, K, Ca, Mg) were concentrated in the biochar and were significantly higher in the biochars produced at 500°C.
A large proportion of N was conserved in the biochar, ranging from 27.4% in the PL biochar to 89.6% in the PC biochar. The
amount of N conserved was inversely proportional to the feedstock N concentration. The cation exchange capacity was
significantly higher in biochar produced at lower temperature. The results indicate that, depending on feedstock, some
biochars have potential to serve as nutrient sources as well as sequester C.
Keywords. Agricultural residues, Biochar, Bioenergy, Black carbon, Carbon sequestration, Charcoal, Plant nutrition,
Pyrolysis, Soil fertility, Soil organic carbon.

P
yrolysis of crop residues to produce renewable ener‐
gy is one option to reduce the use of fossil fuels. Py‐
rolysis generates biochar, oil, and gas products that
can all be used as fuels (Ioannidou and Zabaniotou,
2007). Pyrolytic biochar can also potentially be used as a low‐
cost sorbent (Ioannidou and Zabaniotou, 2007) or as a soil
amendment to improve soil fertility and sequester carbon
(Lehmann et al., 2006; Steiner, 2007). Removal of crop resi‐
dues for energy production can have deleterious effects on
soil organic carbon (SOC) and consequently on soil fertility
(Lal, 2004). Pyrolysis of crop residues with C returned to the
soil in the form of biochar may help maintain or increase
stable SOC pools and cycle nutrients back into agricultural
fields. Pyrolysis with biochar C sequestration may offer an
option to reduce the conflict between cultivating crops for
different purposes, e.g., energy vs. C sequestration or food.
There are several lines of evidence that charcoal plays an
important role in soil fertility. Charcoal has been identified
as an important soil constituent in fertile Chernozems
(Schmitdt et al., 1999) and in anthropogenic enriched dark
Submitted for review in August 2008 as manuscript number SW 7634;
approved for publication by the Soil & Water Division of ASABE in
November 2008.
The authors are Julia W. Gaskin, Sustainable Agriculture Coordinator,
Christoph Steiner, Post‐Doctoral Associate, Keith Harris, Technician, K.
C. Das, Associate Professor, and Brian Bibens, Research Engineer,
Department of Biological and Agricultural Engineering, Driftmier
Engineering Center, University of Georgia, Athens, Georgia.
Corresponding author: Julia W. Gaskin, Department of Biological and
Agricultural Engineering, Driftmier Engineering Center, University of
Georgia, Athens, GA 30602; phone: 706‐542‐1401; fax: 706‐542‐1886;
e‐mail: jgaskin@engr.uga.edu.
soil (Terra Preta) found throughout the lowland portion of the
Amazon Basin (Glaser et al., 2000). Research on tropical
soils indicates that charcoal amendments can increase and
sustain soil fertility (Steiner et al., 2007). The beneficial ef‐
fects appear to be related to alterations in soil physical, chem‐
ical, and biological properties, such as reduced acidity
(Topoliantz et al., 2005), increased cation exchange capacity
(CEC) (Cheng et al., 2008; Liang et al., 2006), enhanced ni‐
trogen (N) retention (Lehmann et al., 2003; Steiner et al.,
2008b), increased microbiological activity (Steiner et al.,
2008a), and increased mycorrhizal associations (Warnock et
al., 2007). Research on the effect of wildfire charcoal in for‐
est ecosystems indicates that it stimulates microbial activity
(Pietikäinen et al., 2000) and influences nitrogen cycling
(Berglund et al., 2004; DeLuca et al., 2006; Wardle et al.,
1998). Research also indicates that charcoal is recalcitrant
(Seiler and Crutzen, 1980), and it may persist for hundreds or
thousands of years.
Charcoals produced from wildfire or traditional charcoal
production may have different chemical and physical charac‐
teristics from pyrolytic biochars created under specific con‐
ditions for energy production. Both feedstock and pyrolysis
conditions such as temperature and carrier gas affect the
chemical and physical characteristics of biochar (Antal and
Grønli, 2003; Bansal et al., 1988; Benaddi et al., 2000; Guo
and Rockstraw, 2007a; Strelko et al., 2002). Most of the liter‐
ature discusses high‐temperature biochars that are produced
at greater than 500°C or activated carbon typically produced
at 800°C. As pyrolysis temperatures increase, volatile com‐
pounds in the biochar matrix are lost, surface area and ash in‐
crease, but surface functional groups that can provide
exchange capacity decrease (Guo and Rockstraw, 2007a).

Transactions of the ASABE

Vol. 51(6): 2061-2069 E 2008 American Society of Agricultural and Biological Engineers ISSN 0001-2351 2061
 Pyrolysis of nutrient‐rich feedstock is likely to produce
nutrient‐rich biochar, but nutrient conservation and availabil‐
ity in biochars is not well understood. Nutrients susceptible
to volatilization such as N are almost completely lost after a
burn (Giardina et al., 2000). Whether elements are retained
during pyrolysis, the availability of nutrients for plants, and
the effect of pyrolysis conditions on these characteristics are
unclear. For biochar to be used in agriculture, a better under‐
standing of its properties and how it affects soil fertility is
needed. Therefore, our objectives were to determine the ef‐
fect of feedstock, temperature, and carrier gas on key charac‐
teristics of biochar for agricultural use. Specifically, we
wished to compare characteristics critical for agricultural use
including pH, CEC, total nutrient concentrations, and poten‐
tially available nutrient concentrations in biochars from three
feedstocks under two temperature regimes using two carrier
gases with and without secondary steam activation.
MATERIALS AND METHODS
BIOCHAR PRODUCTION
We selected three common feedstocks to represent a range
of physical properties and mineral content: raw poultry litter
from broiler houses (Gallus domesticus, PL), pelletized pea‐
nut hulls (Arachis hypogaea, PN), and raw pine chips (Pinus
taeda, PC). Biochars were produced in a batch pyrolysis unit
at two peak temperatures (400°C and 500°C) with either
steam or nitrogen (N2) as a carrier gas. The biochars produced
with N2 as a carrier gas were produced with or without steam
activation at the original pyrolysis temperatures (400°C and
500°C). Each of the production combinations (three feed‐
stocks, three pyrolysis types, two temperatures = 18) was rep‐
licated three times. The conversion efficiency was calculated
as the percentage of the feedstock input (dry weight, DW) and
biochar output (biochar DW / feedstock DW).
CHEMICAL ANALYSES
Biochars were ground in a ball mill to pass a 300 mm sieve
before nutrient analysis. Feedstock and the biochars were
analyzed for total C, N, and sulfur (S) by dry combustion
(CNS‐2000, Leco Corp., St. Joseph, Mich.). Total minerals
were extracted using a closed‐vessel microwave digestion
with HNO3 (USEPA method 3050; USEPA, 1994). A Meh‐
lich I extraction (0.05 M HCl + 0.0125 M H2SO4) (Mehlich,
1953) was also used on biochar samples as an index of poten‐
tially plant‐available nutrients. Aluminum, Cu, Ca, Fe, Mg,
Mn, P, K, Na, and Zn were measured by inductively coupled
plasma spectrometry (ICP, Thermo Jarrell‐Ash model 61E,
Thermo Fisher Scientific, Waltham, Mass.).
Biochar pH was measured in deionized water using a 1 to
5 wt/wt ratio. Samples were thoroughly mixed and allowed
to equilibrate for 1 h. The pH was measured with a digital pH
meter (AR15, Thermo Fisher Scientific, Waltham, Mass.).
Cation exchange capacity of the biochar was measured by
a modified ammonium‐acetate compulsory displacement
Table 1. Total element concentrations in the three agricultural feedstocks used for pyrolysis at 400°C and 500°C.
Values in g kg‐1
Feedstock C N P K S Ca Mg
Poultry litter (PL) 326 45.1 19.5 29.5 5.8 28.0 5.66
Peanut hulls (PN) 552 13.6 0.61 5.06 0.9 1.84 0.79
Pine chips (PC) 571 0.9 0.08 0.59 0 0.75 0.21
2062
(Sumner and Miller, 1996). Samples were leached with de‐
ionized water five times before starting the CEC extraction
to reduce interference from soluble salts. Twenty mL of de‐
ionized water was added to a 1 g sample of biochar in a dis‐
posable nalgene 0.45 mm cellulose nitrile filter flask. The
flask was placed on an orbital shaker and shaken at 180 rpm
for 5 minutes. The sample was vacuum filtered, and the lea‐
chate was saved for further analysis. After the fifth wash,
10mL of Na‐acetate (pH 7) was added to the sample, and the
mixture shaken for 10 min. This process was repeated three
times to ensure that exchange sites were saturated with Na
ions. Biochar samples were then washed three times with
ethanol to remove excess Na. Sodium ions were displaced
with NH4‐acetate (pH 7) three times and measured by atomic
adsorption (PE 4100ZL, Perkin Elmer, Waltham, Mass.).
The reserved leachate from the five washings (CEC proce‐
dure above) was composited and analyzed for dissolved car‐
bon (DC), dissolved inorganic carbon (DIC), ammonium‐
nitrogen (NH4-N), and nitrate‐nitrogen (NO3-N). Dissolved
carbon and DIC was measured by combustion (Shimadzu
TOC‐5050A, Shimadzu, Columbia, Md.). Dissolved organic
C (DOC) was calculated by difference (DOC = DC - DIC).
Nitrate‐nitrogen and NH4-N were analyzed on an
autoanalyzer using cadmium reduction and phenate
colorimetric methods (EnviroFlow 3000, Perstorp, Toledo,
Ohio).
STATISTICAL ANALYSES
Treatment effects were analyzed by general linear model
(GLM) univariate analysis of variance (ANOVA). The
detection limit was used for results below the detection limit,
if other results were above the limit. This allowed a
conservative estimate of the elemental concentration of the
biochar. If all results were below the detection limit, then no
statistical analysis was performed. Significant differences
(p< 0.05) between the feedstock and treatments were
separated by the Tukey test. Statistical analyses and plots
were performed using SPSS 12.0 and SigmaPlot 8.02 (SPSS,
Inc., Chicago, Ill.).
RESULTS AND DISCUSSION
Steam pyrolysis of the peanut hull pellets in the batch reactor
presented difficulties due to excessive swelling by the peanut
hull feedstock that clogged the reactor. Low‐temperature steam
pyrolysis in a batch reactor may not be appropriate for this
feedstock. Analysis of PC and PL biochars revealed no
difference in total nutrients, Mehlich I extractable nutrients,
CEC, or pH between steam and N2 as carrier gas; consequently,
we report on the results from pyrolysis with the N2 carrier gas
with or without subsequent steam activation.
INFLUENCE OF FEEDSTOCK
The total element concentrations in the feedstock had the
strongest influence on the chemical composition of the
Values in mg kg‐1
Al Fe Na Cd Cr Cu Mn B Mo Ni Zn
6.32 3.91 9.27 1.4 7.3 381 377.0 49.9 3.5 8.0 414
0.92 0.42 0.04 <1 2.0 36.5 44.0 15.2 15.6 1.5 20.2
0.03 0.13 0.04 <1 2.1 1.7 13.8 2.1 <1.0 <2.0 47.8
TRANSACTIONS OF THE ASABE
 Table 2a. Means and standard errors for pH, CEC, and total macronutrient concentrations in poultry litter, peanut
hull, and pine chip biochars. Feedstock and temperature columns indicate significant differences (p < 0.05, n = 3).[a]
Poultry Litter (PL) Peanut Hulls (PN) Pine Chips (PC)
400°C 500°C 400°C 500°C 400°C 500°C
Biochar SA Biochar SA Biochar SA Biochar SA Biochar SA Biochar SA Feedstock
pH 10.1 10.1 9.74 9.88 10.5 10.5 10.1 9.96 7.55 7.99 8.30 8.10
(S.U.) ±0.04 ±0.07 ±0.05 ±0.09 ±0.05 ±0.10 ±0.02 ±0.01 ±0.09 ±0.09 ±0.15 ±0.60 PC<PL<PN Temp.
CEC 61.1 57.4 38.3 37.0 14.2 11.7 4.63 4.46 7.27 6.00 5.03 6.02
(cmol kg‐1) ±0.73 ±1.4 ±1.7 ±1.6 ±0.46 ±2.04 ±0.10 ±0.13 ±0.54 ±0.11 ±0.85 ±3.12 PC<PN<PL 500<400
C 392 399 392 421 732 762 804 806 739 761 817 820
(g kg‐1) ±3.8 ±7.4 ±8.6 ±23 ±14 ±3.4 ±1.7 ±5.8 ±17 ±3.6 ±1.9 ±17 PL<PN, PC 400<500
N 34.7 34.7 30.9 32.3 24.3 24.0 24.8 24.8 2.55 1.95 2.23 2.20
(g kg‐1) ±0.79 ±0.77 ±0.89 ±1.6 ±0.18 ±0.37 ±0.89 ±0.34 ±0.40 ±0.06 ±0.09 ±0.12 PC<PN<PL 500<400
P 30.1 32.2 35.9 34.8 1.83 1.70 1.97 2.06 0.15 0.14 0.14 0.20
(g kg‐1) ±0.16 ±2.3 ±1.6 ±2.6 ±0.11 ±0.12 ±0.03 ±0.11 ±0.004 ±0.004 ±0.02 ±0.02 PC<PN<PL 400<500
K 51.1 52.6 58.6 54.7 15.2 14.40 16.4 16.5 1.45 1.51 1.45 2.25
(g kg‐1) ±1.3 ±4.9 ±2.9 ±1.5 ±0.58 ±1.40 ±0.19 ±0.79 ±0.06 ±0.07 ±0.18 ±0.25 PC<PN<PL 400<500
Ca 42.7 45.7 50.4 49.1 4.62 4.46 5.12 5.21 1.71 1.69 1.85 2.17
(g kg‐1) ±0.30 ±3.0 ±2.2 ±3.7 ±0.06 ±0.29 ±0.12 ±0.20 ±0.11 ±0.02 ±0.14 ±0.04 PC<PN<PL 400<500
Mg 10.7 11.4 12.9 12.4 2.19 2.17 2.50 2.59 0.60 0.58 0.59 0.76
(g kg‐1) ±0.23 ±0.91 ±0.50 ±1.0 ±0.06 ±0.16 ±0.05 ±0.11 ±0.04 ±0.03 ±0.06 ±0.01 PC<PN<PL 400<500
S 13.67 12.3 13.93 13.9 0.56 0.51 0.55 0.37 0.01 0.16 0.06 0.08
(g kg‐1) ±0.39 ±0.09 ±1.1 ±0.37 ±0.02 ±0.03 ±0.09 ±0.09 ±0.04 ±0.05 ±0.01 ±0.04 PC, PN<PL
[a] SA = steam activation.

Table 2b. Means and standard errors for total micronutrient and selected element concentrations in poultry litter, peanut
hull, and pine chip biochars. Feedstock and temperature columns indicate significant differences (p < 0.05, n = 3).[a]
Poultry Litter (PL) Peanut Hulls (PN) Pine Chips (PC) Feedstock Temp.
400°C 500°C 400°C 500°C 400°C 500°C
Biochar SA Biochar SA Biochar SA Biochar SA Biochar SA Biochar SA
Al 9.87 8.12 13.02 14.25 2.40 2.33 2.73 2.81 0.07 0.05 0.07 0.06
(g kg‐1) ±1.36 ±1.59 ±0.36 ±1.97 ±0.07 ±0.16 ±0.05 ±0.13 ±0.01 ±0.005 ±0.01 ±0.008 PC, PN<PL 400<500
Fe 6.06 5.55 8.03 8.89 1.00 0.97 1.15 1.20 0.15 0.04 0.05 0.20
(g kg‐1) ±0.52 ±0.42 ±0.55 ±1.27 ±0.02 ±0.07 ±0.02 ±0.06 ±0.11 ±0.007 ±0.01 ±0.07 PC, PN<PL 400<500
Na 15.1 15.8 17.2 16.6 0.026 0.028 0.035 0.044 <0.014 0.053 0.013 0.075
(g kg‐1) ±0.31 ±1.37 ±1.02 ±1.12 ±0.001 ±0.006 ±0.004 ±0.005 ±0.004 ±0.032 ±0.002 ±0.054 PN, PC<PL
B 91.5 96.0 100 93.0 32.5 29.9 33.7 34.1 5.69 6.69 4.21 6.94
(mg kg‐1) ±3.16 ±8.25 ±0.31 ±3.98 ±1.57 ±2.87 ±0.27 ±1.15 ±0.30 ±0.21 ±0.62 ±1.33 PC<PN<PL
Cd 2.75 <2.65 <1.10 <1.35 <1.35
(mg kg‐1) ±0.73 ±0.83 <1 ±0.10 ±0.35 ±0.35 <1 <1 <1 <1 <1 <1
Cr 28.0 28.8 59.4 56.1 3.95 3.00 3.63 3.94 1.23 3.43 17.7
(mg kg‐1) ±4.1 ±5.2 ±3.3 ±4.5 ±0.31 ±0.57 ±0.31 ±0.16 <1 ±0.09 ±0.88 ±5.9 PN, PC<PL 400<500
Cu 805 880 1034 943 16 13 19 19 25 10 9 13
(mg kg‐1) ±23 ±49 ±68 ±81 ±0.60 ±1.27 ±0.50 ±1.84 ±7.03 ±6.18 ±2.34 ±5.57 PC, PN<PL 400<500
Mn 596 637 725 697 116 116 131 136 274 269 258 350
(mg kg‐1) ±5.6 ±37 ±29 ±46 ±2.3 ±8.0 ±2.3 ±5.7 ±9.3 ±7.8 ±30 ±4.0 PN<PC<PL 400<500
Mo 17.1 12.1 14.2 13.8 4.78 <4.11
(mg kg‐1) ±5.3 ±0.41 ±1.1 ±1.2 ±3.6 <1 <1 <1 <1 <1 <1 ±3.11 PC, PN<PL
Ni 13.6 19.5 20.3 29.1 <2.29 <2 <2 <10.4 <2 <2 <2.91 17.5
(mg kg‐1) ±0.00 ±3.7 ±1.1 ±8.4 ±0.29 ±0 ±0 ±8.0 ±0 ±0 ±0.55 ±14.7 PC, PN<PL
Zn 628 680 752 728 35 31 37 36 15 16 18 20
(mg kg‐1) ±12 ±41 ±28 ±50 ±2.2 ±2.9 ±2.1 ±0.00 ±1.1 ±0.7 ±0.6 ±2.4 PC, PN<PL 400<500
[a] SA = steam activation; < indicates mean contains results below the detection limit; ±0.00 indicates all results were near instrument detection limit.

biochar. Concentrations of plant nutrients in the feedstocks
generally followed the pattern of PC < PN < PL. Feedstock
carbon concentrations had the opposite pattern, with PL < PN
< PC (table 1).
There were significant differences in C concentrations in
the biochar, with PL containing less C than the PN or PC
biochar (table 2a). The nutrient‐rich poultry litter contains
relatively more minerals than the other feedstocks, which
decreases the C content. Nitrogen, P, K, Ca, and Mg
concentrations in the biochar were significantly different,
with PC < PN < PL (table2a). The concentration of the
micronutrients B, Cu, Fe, Mn, Na, and Zn were significantly
higher in PL biochar (p < 0.05), but there were no differences
detected between the PN and PC biochars except for Mn
(table 2b). Concentrations of metals such as Al, Cr, Ni, and
Mo were low. The PL biochar contained the highest

Vol. 51(6): 2061-2069 2063

Figure 1. Percentages with standard errors of feedstock nutrients conserved in the biochar and percentages of total nutrients that were Mehlich I
extractable at two pyrolysis temperatures and in three biochars. Letters above the columns indicate significant difference of nutrients conserved
between biochar types (p < 0.05, n = 3). Letters within columns indicate significant difference in the percentage of total nutrients that were MehlichI
extractable (p < 0.05, n = 3).

concentrations of these metals, as would be expected from
the higher feedstock concentrations. Cadmium was below
the detection limit in PC biochars and near or at the detection
limits in PL and PN biochars (table 2b).
The amount of N conserved ranged from 27.4% in the PL
biochar to 89.6% in the PC biochar and was inversely
proportional to the feedstock N concentration (fig. 1b and
table 1). The higher N losses seen from the PL were likely due
to the volatilization of the poultry manure NH4-N and easily
decomposable N-containing organic compounds in the
manure, such as uric acid. In contrast, the low concentration
of N in the PC feedstock is likely to be incorporated into
complex structures that are not easily volatilized.
About 60% of the P in the PL and PC feedstock was
retained in the PL and PC biochar, while nearly 100% of the
P in the PN feedstock was retained in the PN biochar (fig. 1c).
In general, the PL biochar had a lower proportion of nutrients
retained than the PN or the PC biochar (figs. 1c through 1f).
This may be due to a higher proportion of some of these
elements retained in the aqueous/bio‐oil fraction in PL
biochar (K. C. Das, 2007, unpublished data, University of
Georgia, Athens, Ga.).
The pattern of Mehlich I extractable concentrations was
similar to that of the total nutrient concentrations (tables 3a
and 3b). There were significant differences in Mehlich I
extractable P, K, Ca, and Mg concentrations, with PC < PN
< PL. There were differences by feedstock in the percentage
of the total nutrients that were Mehlich I extractable (figs. 1a
through 1f). Only 19% of the PL biochar P was Mehlich I
extractable compared to over 40% in the PN biochar (400°C,
fig. 1c). About 90% of the PL biochar K was Mehlich I
extractable compared to only 45% in the PN biochar (400°C,

2064 TRANSACTIONS OF THE ASABE

 Table 3a. Means and standard errors of the Mehlich I macronutrient concentrations in poultry litter, peanut hull,
and pine chip biochars. Feedstock and temperature columns indicate significant differences (p < 0.05, n = 3).[a]
Poultry Litter (PL) Peanut Hulls (PN) Pine Chips (PC) Feedstock Temp.
400°C 500°C 400°C 500°C 400°C 500°C
Biochar SA Biochar SA Biochar SA Biochar SA Biochar SA Biochar SA
P 5.58 4.09 5.33 4.66 0.76 0.67 0.57 0.59 0.03 0.034 0.04 0.06
(g kg‐1) ±0.31 ±1.22 ±0.18 ±0.20 ±0.02 ±0.06 ±0.04 ±0.03 ±0.002 ±0.004 ±0.008 ±0.03 PC<PN<PL
K 46.2 34.1 38.1 40.0 6.84 6.28 5.91 6.76 0.30 0.38 0.41 0.97
(g kg‐1) ±0.96 ±8.40 ±2.68 ±2.81 ±0.16 ±0.67 ±0.28 ±0.30 ±0.009 ±0.02 ±0.06 ±0.32 PC<PN<PL
Ca 3.34 1.95 2.21 1.63 1.68 1.48 1.19 1.22 0.30 0.31 0.43 0.39
(g kg‐1) ±0.84 ±0.82 ±0.36 ±0.13 ±0.02 ±0.15 ±0.06 ±0.06 ±0.04 ±0.05 ±0.10 ±0.16 PC<PN<PL
Mg 3.09 2.19 3.03 2.92 0.80 0.62 0.37 0.39 0.05 0.06 0.06 0.08
(g kg‐1) ±0.28 ±0.68 ±0.13 ±0.05 ±0.03 ±0.09 ±0.02 ±0.01 ±0.008 ±0.009 ±0.01 ±0.04 PC<PN<PL
[a] SA = steam activation.

Table 3b. Means and standard errors of the Mehlich I micronutrient and selected element concentrations in poultry litter,
peanut hull, and pine chip biochars. Feedstock and temperature columns indicate significant differences (p < 0.05, n = 3).[a]
Poultry Litter (PL) Peanut Hulls (PN) Pine Hips (PC) Feedstock Temp.
400°C 500°C 400°C 500°C 400°C 500°C
Biochar SA Biochar SA Biochar SA Biochar SA Biochar SA Biochar SA
Al 0.47 11.3 1.43 1.52 330 585 1129 1360 6.17 6.35 7.53 12.12
(g kg‐1) ±0.11 ±2.24 ±0.31 ±0.24 ±24 ±213 ±32 ±51 ±0.59 ±0.53 ±1.12 ±4.76 PL, PC<PN 400<500
Fe 0.66 3.16 0.06 0.19 140 142 197 221 3.72 4.58 14.6 33.3
(g kg‐1) ±0.21 ±0.08 ±0.007 n=1 ±2.2 ±18 ±3.8 ±11 ±0.26 ±0.19 ±2.89 ±8.39 PL, PC<PN 400<500
Na 9.57 7.08 6.98 7.24 0.02 0.02 0.02 0.03 0.03 0.03 0.03 0.08
(g kg‐1) ±0.19 ±1.61 ±0.43 ±0.07 ±0.001 ±0.002 ±0.58 ±0.002 ±0.002 ±0.002 ±0.005 ±0.037 PN, PC<PL
B 16.7 18.8 18.67 20.4 4.20 4.96 3.97 5.84 0.45 0.41 0.52 1.15
(mg kg‐1) ±1.59 ±2.05 ±0.85 ±0.59 ±0.12 ±1.08 ±0.22 ±0.66 ±0.04 ±0.07 ±0.07 ±0.49 PC<PN<PL
Cr 0.19 0.19 0.14 0.11 0.41 0.52
(mg kg‐1) ±0.03 ±0.02 ±0.01 ±0.01 <0.04 <0.04 ±0.04 ±0.03 <0.06 <0.06 <0.06 <0.06
Cu 0.40 0.29 <0.08 <0.05 0.67 <0.59 <0.04 6.55 2.48 2.70 3.82
(mg kg‐1) ±0.06 ±0.08 ±0.02 ±0.005 ±0.04 ±0.32 ±0.001 <0.04 ±2.18 ±0.68 ±0.89 ±3.34 PL, PN<PC
Mn 7.69 8.64 6.75 5.17 24.7 21.2 14.4 16.3 22.6 25.2 24.1 36.2
(mg kg‐1) ±1.23 ±1.28 ±1.03 ±0.43 ±0.70 ±2.33 ±0.45 ±0.71 ±2.61 ±3.79 ±6.67 ±12.5 PL<PN<PC
Mo 0.87 1.11 1.42 1.94 0.15 0.25 0.11 0.65
(mg kg‐1) ±0.19 ±0.22 ±0.14 ±0.19 <0.04 <0.04 <0.04 <0.04 ±0.002 ±0.06 ±0.03 ±0.35
Ni
(mg kg‐1) <0.08 <0.08 <0.08 <0.08 <0.08 <0.08 <0.08 <0.08 <0.2 <0.2 <0.2 <0.2
Zn 0.06 0.30 0.05 10.51 7.36 5.58 6.30 2.20 2.31 1.36 3.66
(mg kg‐1) ±0.01 ±0.02 ±0.07 <0.04 ±1.22 ±0.59 ±0.26 ±0.45 ±0.17 ±0.21 ±0.26 ±1.17 PL<PC<PN
[a] SA = steam activation; < indicates mean contains results below the detection limit.

fig. 1d). Manganese and Zn concentrations were significantly
lower in the PL biochar than the PC or PN biochars. Copper,
Al, and Fe was also lower in the PL biochar compared to the
PN biochar. These patterns are the reverse of that seen in the
feedstock or the total element concentrations in the biochars.
If pyrolysis can reduce P and other metal availability in
poultry litter, it may reduce some of the environmental
concerns associated with land application of poultry litter.
These results should be interpreted with caution. The
Mehlich I extraction, which is a weak double acid extraction,
may not have been strong enough to remove these acid‐
soluble cations under the high pH conditions found in the PL
biochar.
The Mehlich I extraction was developed for acidic soils in
the southeastern U.S. with low CEC or base saturation (Kuo,
1996), and it is the standard extraction used for plant‐
available nutrients and fertilizer recommendations in
Alabama, Georgia, Florida, South Carolina, Tennessee, and
Virginia. In this study, Mehlich I extractable element
concentrations were used as an index to compare the
potential for different biomass sources and production
techniques to supply plant‐available nutrients. The
extraction has not been calibrated for biochar and may not
reflect actual plant‐available nutrient concentrations.
However, data from a greenhouse trial using pine chip and
peanut hull biochar amendment of three different Ultisols
(Speir, 2008) and from a field trial with the same biochars
(Gaskin et al., 2007) indicate an increase in Mehlich I K and
Mg in soils amended with peanut hull biochar. The increased
Mehlich I K in the soil was reflected in an increase of these
nutrients in corn tissue (Zea mays) in the field trial.
The pH and CEC of the biochars were also significantly
influenced by feedstock (table 2a). All the biochars were
basic, with the highest pH seen in the PN biochar. Tryon
(1948) reported increased soil pH with the addition of pine
and hardwood charcoal. He attributed the greater pH increase
seen in the hardwood charcoal treatment to the higher ash
content, in particular to the hydrolysis of salts of Ca, K, and
Mg in the presence of water. In this study, PC biochar had
both the lowest total concentrations of these cations and the

Vol. 51(6): 2061-2069 2065

 Table 4. Means and standard errors of dissolved carbon (DC), dissolved inorganic carbon (DIC), dissolved organic carbon
(DOC), ammonium‐nitrogen (NH4-N), and nitrate‐nitrogen (NO3-N) in leachate from poultry litter, peanut hull,
and pine chip biochars. Feedstock and temperature columns indicate significant differences (p < 0.05, n = 3).[a]
Poultry Litter (PL) Peanut Hulls (PN) Pine Chips (PC) Feedstock Temp.
400°C 500°C 400°C 500°C 400°C 500°C
Biochar SA Biochar SA Biochar SA Biochar SA Biochar SA Biochar SA
DC 2.20 1.85 0.85 0.75 0.51 0.40 0.52 0.41 0.13 0.13 0.12 0.19
(g kg‐1) ±0.31 ±0.25 ±0.01 ±0.02 ±0.09 ±0.02 ±0.02 ±0.10 ±0.10 ±0.10 ±0.05 ±0.07 PC<PN<PL 500<400
DIC 0.39 0.44 0.57 0.54 0.32 0.31 0.38 0.37 0.014 0.025 0.034 0.055
(g kg‐1) ±0.04 ±0.04 ±0.03 ±0.03 ±0.03 ±0.03 ±0.03 ±0.04 ±0.003 ±0.003 ±0.005 ±0.012 PC<PN<PL 400<500
DOC 1.81 1.46 0.28 0.21 0.20 0.10 0.14 0.10 0.12 0.10 0.09 0.10
(g kg‐1) ±0.34 ±0.29 ±0.04 ±0.04 ±0.06 ±0.03 ±0.03 ±0.03 ±0.01 ±0.01 ±0.003 ±0.01 PC, PN<PL 500<400
NH4‐N 8.5 6.69 11.3 3.49 2.86 1.94 2.12 2.28 1.75 7.93 2.41 2.37
(mg kg‐1) ±0.39 ±0.69 ±6.41 ±0.15 ±0.27 ±0.01 ±0.12 ±0.44 ±0.29 ±6.16 ±0.16 ±0.08 PN<PL
NO3‐N <0.4 <0.4 <0.4 <0.4 1.02 1.07 1.27 1.11 <0.4 <0.4 <0.4 <0.4
(mg kg‐1) ±0.01 ±0.02 ±0.06 ±0.02
[a] SA = steam activation; < indicates mean contains results below the detection limit.

lowest pH, which would support Tyron's hypothesis;
however, the pH of PL biochar was similar to the PN biochar
although it contained higher concentrations of total Ca, K,
and Mg than PN biochar (table 2).
Higher CEC was associated with higher concentrations of
minerals in the feedstock. Mészáros et al. (2007)
hypothesized that K, Mg, Na, and P in the biomass may
catalyze the formation of oxygen groups on the biochar
surface at low pyrolysis temperatures. Oxygen groups such
as carboxyls, lactones, and phenols could contribute to the
presence of negative surface charges (Boehm, 1994).
Dissolved C concentrations were very low (table 4).
Feedstock had a significant effect on DC in the biochar
leachate, with PC < PN < PL. Dissolved inorganic C was also
affected by feedstock, with PC < PN < PL. The PL biochar
had a higher proportion of DOC than PC or PN biochars. The
PL feedstock is a combination of wood chip (typically pine)
bedding and poultry manure. The manure may contribute to
higher DOC leached from the PL biochar.
DOC plays an important role in many soil processes,
including serving as an energy source for the microbial
community and reacting with other soil solution components
(Sposito, 1989). Biochars are known to contain condensed
volatile compounds. These compounds are either lost to the
gaseous or liquid phase or undergo further reactions to form
secondary char at high temperatures (Antal and Grønli,
2003). Garcia‐Perez et al. (2007) identified water‐soluble
compounds from pyrolysis of lignin materials to contain
mono‐ and oligo‐sugars, formic and aecetic acids, as well as
methanol, hydroxyl‐acetaaldehyde, and 1‐hydroxyl‐2‐
propanone. Schnitzer et al. (2007) identified numerous
organic compounds in the light and heavy fractions of poultry
litter pyrolyzed at 330°C, including N-heterocyclics,
substituted furans, phenol and substituted phenols, benzenes
and substituted benzenes, as well as aliphatic C chains. It is
likely that some of these compounds remain trapped in the
biochar pore structure, but few of these compounds appear to
be immediately water soluble.
Ammonium‐nitrogen in the biochar leachate was also
found in very low concentrations (table 4). No NO3-N was
detected in any of the leachates. The NH4-N concentrations
were highest in the leachate from the PL biochar. Fresh
poultry litter typically contains about 2.8 g NH4-N kg-1
(University of Georgia Agricultural and Environmental
Services Laboratory, unpublished data). Small amounts of
this NH4-N may remain trapped, or microbes may have
mineralized nitrogen‐containing organic compounds in the
biochar. Das et al. (2008) found that the liquid products
obtained from poultry litter pyrolysis enhanced microbial
growth in well water and concluded that N-heterocyclic
compounds derived from proteins were responsible for that
increase. A small fraction of these compounds may be
present in the biochars.
INFLUENCE OF PYROLYSIS TEMPERATURE
The average conversion ratio (biochar weight / feedstock
weight) was 33.2%. Biochar yield decreased with increased
pyrolysis temperature, and except for N, nutrient
concentrations were higher in the biochar produced at 500°C
(tables 2a andb). Due to the wide range of initial nutrient
concentrations in the feedstock, there were significant
interactions between feedstock and temperature for total N,
P, Mg, Mn, Cu, Fe, Zn, and Al (p < 0.05).
As noted earlier, N was conserved in the biochar (fig. 1b).
After forest fires, on average, only 3% of the N in the biomass
is found in ash, which contains black carbon or biochar
(Giardina et al., 2000). Almendros et al. (2003) found C and
N enrichment in charred residues during thermal
transformation of peat organic matter. Nitrogen was
incorporated into structures resistant to heating at moderate
thermal oxidation by aromatization and formation of
heterocyclic N (Almendros et al., 2003). Studies of wildfire
effects on biomass composition indicate that N begins to
volatilize at 200°C, and above 500°C half of the N in organic
matter is lost to the atmosphere. Our study indicated that a
relatively high proportion of the feedstock N was conserved
at low pyrolysis temperatures, and as expected more N was
retained in the biochar at 400°C compared to 500°C (fig. 1b).
Knicker et al. (2005) has shown that fire and carbonization
can increase the N content of SOC, but the alterations in
chemical structure have long‐term consequences for N
availability (Knicker and Skjemstad, 2000). Field trials of PN
and PC biochar as a soil amendment with corn (Zea mays)
indicate that PN biochar N is not plant available (Gaskin et al.,
2007). However, Tagoe et al. (2008) studied N recovery of
15N‐labeled chicken manure and did not find differences in N
availability between carbonized and dried chicken manure.

2066 TRANSACTIONS OF THE ASABE


Figure 2. Representative relationship of the ratio of nutrient (K) in the
biochar to feedstock and conversion efficiency for pine chip, peanut hull,
and poultry litter biochars. Solid circles represent means, and bars
indicate standard errors.
The total concentration of other elements (P, K, Ca, and
Mg) significantly increased with increasing volatization
losses of C, H, O, and N (tables 2a and 2b). Potassium is
representative of the nutrient concentration seen (fig. 2).
Potassium and P vaporize at temperatures above 760°C, S
and Na need temperatures above 800°C, and Mg and Ca are
lost only at temperatures above 1107°C and 1240°C,
respectively (Lide, 2004, reviewed by Knicker, 2007). There
was a significant interaction between temperature and
feedstock for Mehlich I extractable concentrations of these
elements (p = 0.05). At the low nutrient concentrations seen
in the PC biochar, temperature appeared to have little effect.
In the PN and PL biochars, Mehlich I extractable nutrients
tended to decrease with increasing temperature. Mehlich I
extractable Al and Fe were significantly increased in the
500°C biochar (table 3b).
Vol. 51(6): 2061-2069
The CEC of biochar produced at 500°C was significantly
less than that produced at 400°C (table 2a, p < 0.01). There
was a significant interaction between feedstock and
temperature. In general, the literature indicates the loss of
surface functional groups with the increase in pyrolysis
temperature. Guo and Rockstraw (2007b) showed that the
number of acidic functional groups decreased with
increasing temperature. The highest decrease occurred
between 300°C and 400°C, and the loss of these acidic
groups slowed after 400°C. This process may have
contributed to the lower CEC seen at higher temperatures.
Iyobe et al. (2004) indicated that lignin and cellulose undergo
thermolysis at 400°C to 500°C, which creates acidic
functional groups such as carboxyls and phenolic hydroxyls.
Chun et al. (2004) found decreasing acidity and increasing
basicity with increasing pyrolysis temperature.
Temperature influenced DC (table 4). The higher temper-
ature reduced the concentration of organic C but increased
inorganic C significantly.
INFLUENCE OF STEAM ACTIVATION
Steam activation had little effect on the studied
parameters (tables 2a, 2b, 3a, 3b, and 4). Production
technology is known to influence physical parameters, and
steam can improve the yield and surface characteristics at
elevated pressures and temperatures (Antal and Grønli,
2003). At the relatively low pyrolysis temperatures used in
this study, we only found significantly higher C and Mehlich
I extractable B concentrations in steam‐activated biochar
(p< 0.05).
CONCLUSIONS
Pyrolytic biochar has the potential to be used in
agricultural production to sequester carbon and serve as a
fertilizer. Although pyrolysis conditions are known to affect
the chemical and physical characteristics of biochar, at the
relatively low pyrolysis temperatures used in this study,
feedstock characteristics had the greatest influence on key
agricultural characteristics. Carbon concentrations in the
biochars decreased with increasing mineral content of the
feedstock. Little DC was leachable from the fresh biochar. A
high proportion of the feedstock N was conserved in the
biochar; however, the N may not be plant available. Nutrients
such as P, K, and Ca are extractable with a weak double acid
extractant and may be plant available.
The higher pyrolysis temperature increased nutrient
concentrations, except for N, but decreased CEC. Recent
literature has shown that natural long‐term oxidation of
biochar in the soil increases the amount of negative charges
on the biochar surface (Cheng et al., 2008). Development and
optimization of pyrolysis and post‐production treatments to
increase CEC or available nutrients is important in order to
increase the immediate benefits of biochar applications in
agriculture.
ACKNOWLEDGEMENTS
This work was conducted with funding from the State of
Georgia and the U.S. Department of Energy. We wish to
thank Dr. Jim Kastner for his helpful comments on various
ideas in the manuscript, and Mr. Roger Hilten for assistance
with this project.
2067
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Chapter _

CHARACTERIZATION OF CHAR FOR AGRICULTURAL USE IN THE SOILS

OF THE SOUTHEASTERN UNITED STATES
J. GASKIN, K. DAS, A. TASISTRO, L. SONON, K. HARRIS, B. HAWKINS
Dept. of Biological and Agricultural Engineering and Soil, Plant and Water Laboratory, University of Georgia, and EPRIDA., Athens, Georgia, US

Abstract: Char produced from the pyrolysis of biomass has potential as an agricultural amendment for increasing agricultural productivity in the
infertile, low C soils of the southeastern United States, but it is unclear if the chars produced by pyrolysis plants will function in soils
similar to the charcoal in Terra Preta. Recent work in characterizing pyrolysis char indicates that the feedstock, temperature, and
carrier gas has a strong influence on important characteristics for agricultural production, such as available nutrients, cation exchange
capacity, and mineralization rates. Preliminary data indicate pyrolysis char may increase CEC, sorbs P, and serves as a source of plant
available K. Nitrogen mineralization data and first growing season field trials with corn (Zea mays) indicate although some
mineralization may occur, N in high N char (2%) is not readily available. A better understanding of char effects on soil processes is
needed.

Keywords: char, Ultisols, agricultural amendment, nutrients, nitrogen, mineralization, cation exchange capacity, phosphorus sorption

1. INTRODUCTION
Char produced from the pyrolysis of biomass has potential as an agricultural amendment in low fertility soils. Much
of the interest in its potential use as an agricultural amendment has been stimulated by research discussed in this book and
the previous volumes on the role of charcoal in Terra Preta soils. Results from studies conducted in South American and
African tropics on acidic, highly-weathered Oxisols with low organic carbon, cation exchange capacity, and base
saturation indicates that addition of charcoal has significantly influenced nutrient cycling, soil biology, and crop
productivity (Glaser et al. 2002; Lehmann and Rondon 2006; Oguntunde et al. 2004). Increased yields and biomass have
been reported for various legumes (Iswaran et al. 1980; Lehman et al. 2003; Topoliantz et al. 2005) and for corn
(Lehmann and Rondon 2006; Oguntunde et al. 2004). Increased productivity may be related to available nutrients (Glaser
et al. 2002; Lehman et al. 2003; Steiner et al. 2007), or increases in pH (Topoliantz et al. 2005; Steiner et al. 2007), and
cation exchange capacity (CEC)(Steiner et al. 2007, Liang et al. 2006), as well as changes in water relations and soil
biology (Glaser et al. 2002; Steiner et al. 2004). Although most studies report increased plant productivity with charcoal
addition, plant biomass decreases have been observed, particularly at high application rates (Glaser et al. 2002). These
responses could be related to nitrogen immobilization through high C:N ratios and sorption of NH4-N and NO3-
N(Lehmann and Rondon 2006).
The southeastern United States is an important agricultural area. The state of Georgia alone has approximately 4.3
million hectares of corn (Zea mays), soybean (Glycine max), cotton (Gossypium hirsutum), and peanuts (Arachis
hypogaea) in production and 9.6 million hectares of forestland largely in loblolly pine (Pinus taeda) production (USDA
2002; Georgia Forestry Association 2007). The growing interest in biofuels is increasing demands on row crop
production and may also increase demand on forestlands. The Ultisols of the southeastern United States are similar to
tropical Oxisols with low organic carbon contents of less than 1%, low cation exchange capacities of approximately 5
cmol kg-1, and low base saturation of usually less than 30% (Perkins 1987). Char produced as a byproduct of energy
production through pyrolysis may provide an opportunity to increase the productivity of southeastern soils, similar to the
way charcoal functions in Terra Preta. However, because char characteristics vary with feedstock and pyrolysis
conditions (Harris et al. 2006; Antal and Gronli 2003), a better understanding of the influence of these factors on char
characterisitics and the effect of different chars on soil processes in the southeastern United States is needed.

2. PYROLYSIS CONDITIONS AND CHAR CHARACTERISTICS

Char or charcoal corresponds to black carbons that result from the incomplete combustion of biomass. Black carbon
consists of graphite-like planes (graphene layers) that show varying degrees of disorientation. The resulting spaces
between these planes constitute porosity.
The capacity of char to remove impurities from solutions and gases has been known for many centuries. This is due to
the porous nature of the material (Barkauskas 2002) and to the surface chemical properties including the type and number
of functional groups (Stoeckli et al. 2004). Charcoal has chemical reactivity due to the existence of unsaturated valence
(active sites) at the edges of the aromatic planes. The ratio of these active sites in relation to the inert carbon atoms within

1
the graphene layers increases as the surface area increases. Heteroatoms, such as oxygen, hydrogen or nitrogen also have a
strong influence on the mechanisms of the adsorption process. Oxygen is the most important heteroatom and is part of
chemical groups that have both Lewis base properties such as chromene and pyrone or acidic properties such as
anhydrides, lactones, lactols, phenols, carbonyls, and carboxyls. The presence and quantity of these groups affect the
capacity of char to add to the cation or anion exchange capacity of the soil and other soil productivity properties.
Charcoals are known to sorb cations (Lima and Marshall 2005a) and also anions when basic surface oxides are present
due to exposure to the atmosphere (Boehm, 1994). These sorption capacities may create the potential for char to retain
needed nutrients in the low exchange capacity soils of the southeastern United States.

2.1 Feedstocks
The mineral content of the char depends upon the nature of the feedstock and can influence the surface chemistry of
charcoal by interaction with electrons of the aromatic rings and through electron paramagnetism. (Benaddi et al. 2000).
Ash complicates interpretation of surface phenomena of carbon. Ash solubility in water is variable, making the analysis
of surface groups difficult at higher ash concentrations. Insoluble metal oxides involved in the charge development on the
carbon surface become charged in aqueous suspension, and are considered part of the active surface sites.
Feedstock particle size also affects char yield from pyrolysis. At low pyrolysis temperature, larger particle sizes favor
longer inter-pore residence time for volatiles increasing yield (Antal and Gronli, 2003).

2.2 Pyrolysis Conditions

Important parameters that determine quality and yield of the carbonized product are the rate of heating, final
temperature, and the gas environment (Bansal et al., 1988). A low heating rate during pyrolysis leads to lower
volatilization and higher char yields. This creates char with higher carbon contents but does not affect char microporosity.
In addition, chars developed at low heating rates are heavier and denser than those from high heating rates. This may be
an advantage for agricultural use in terms of handling properties.
The final temperature during pyrolysis typically ranges between 400 and 600 oC, and does not exceed 800 oC. Guo and
Rockstraw (2007) observed that surface area and porosity did not develop at temperatures < 300 oC and that from 300 oC
onwards, the concentration of acidic surface groups decreased with increasing temperature. The decrease occurred more
quickly between 300 and 400 oC, and slowed after 400 oC, probably due to an equilibrium between decomposition and
formation of strong acidic surface groups, or because most of the temperature-sensitive strong acidic groups had
disappeared. Iyobe, et al. (2004) reported that thermolysis of cellulose or lignin occurred actively at 400 to 500 oC, with
the formation of acidic functional groups, such as carboxyls and phenolic hydroxyls. The amount of acidic functional
groups continue to decrease with pyrolysis temperatures > 600 oC. Hydroxide (C-OH), C=O, and C-H groups are largely
lost at temperatures > 650°C, and most of the aromatic and C-H groups are decomposed above 750°C (Antal and Gronli,
2003). Above 950°C chars are almost like graphite with little active chemistry on its surfaces. The effect is decreasing
ability to sorb cations. Asada et al. (2002) reported that char obtained by carbonizing bamboo at 500 oC had the highest
removal effect for NH3 compared to carbonizing at 700 or 1000oC. In general, these data indicates chars produced at
lower temperatures (<500 oC) may hold the greatest promise for agricultural use in terms of nutrient holding capacity.
The gas environment during pyrolysis also exerts considerable influence on char properties. Lower carrier gas flow
rates result in longer residence time of vapors in the char matrix, which allows for char-catalyzed secondary reactions to
occur. Steam may increase the presence of oxygen in surface functional groups. Carbon-oxygen surface compounds are
by far the most important in influencing surface reactions, surface behavior, hydrophilicity, and electrical and catalytic
properties of carbons. Substantial quantities of oxygen can be introduced in the course of charcoal production by an
oxidating gas such as steam (Strelko et al. 2002).

2.3 Comparison between traditional two-step pyrolysis and activation with 1-step steam pyrolysis
Typically chars produced at temperatures around 400 to 600 oC do not have the well developed surface areas or
adsorbent properties of activated carbons because of tars deposited on the solid surface that restrict pore structures.
Steam activation at temperatures between 800 and 1,100 oC physically removes these residues and opens pores. After
activation, chars have higher surface area, adsorption capacity and pore size distribution (Gregova et al. 1994; Alaya et al.
2000).
The combination of this two-step process into a single step, which involves pyrolyzing under steam conditions, may
increase surface area and increase adsorbent properties, and requires less energy and less time. Steam pyrolysis at low
temperatures (600 oC) has been shown to increase micropores with the ratio of micropore volume to total pore volume
approaching 95% (Alaya et al. 2000). These authors suggest that steam enhances the evolution of volatile molecules at
2
 lower temperatures and prevents the cracking of volatiles. In addition, gasification (conversion of solids to gases) is
induced much earlier (600 oC compared to 800 to 900 oC in non-steam environments) and results a more porous carbon
skeleton. Chars produced at lower temperatures with steam may have a more active surface chemistry; however, the
literature is not clear about the impacts of one-step processing on nutrient properties and surface chemistry.
Based on available literature on char and activated carbons, it appears that chars produced at low pyrolysis
temperatures with steam may hold the greatest promise for agricultural use due to lower production cost, higher surface
functional groups for sorption, and a more dense char product.

3. AGRICULTURAL CHARACTERISTICS OF PYROLYSIS CHAR

3.1 Feedstock and Pyrolysis Condition Effects on Nutrient Status
Analyses of char from common feedstocks in the Southeastern US confirm the effect of feedstock and temperature on
char composition. Total nutrients were analyzed in chars produced from peanut hull (PN), pine chip (PC), and hardwood
(HW) feedstocks pyrolyzed at low temperatures (380, 400, and 420 oC) with steam in a small furnace, and poultry litter
(PL) at 400 oC in a batch reactor in a steam flow environment (Table 1). At these lower pyrolysis temperatures, the initial
nutrient content of the feedstock had a larger effect on char nutrient concentration than pyrolysis temperature (Table 1,
Figure 1). Small increases in pyrolysis temperature increased the total nutrient concentration in the char of most nutrients
(e.g. K, Figure 1).
Table 1. Total carbon and nutrient concentrations for feedstocks and chars (on an as is basis) produced from those feedstocks at 400 oC with steam.

Constituent Peanut hull Poultry Litter Pine chips Hardwood

Feedstock Char Feedstock* Char Feedstock Char Feedstock Char
C (%) 47.7 65.5 N/A 41.7 46.5 67.0 44.7 70.3
N (%) 1.44 2.00 3.80 3.70 0.05 0.14 0.20 0.30
C:N 33 33 N/A 11 949 543 224 234
S (%) 0.13 0.13 0.42 1.18 0.02 0.02 0.02 0.02
P mg kg -1 732 1,620 11,930 33,580 30.0 235 92.9 278
-1
K mg kg 6,340 15,372 19,339 45,593 436 1,973 937 2,409
Ca mg kg -1 1,880 4,420 17,900 46,760 418 1,686 794 2,709
* Average Georgia poultry litter concentrations analyzed by the University of Georgia Agricultural and Environmental Services Laboratory
N/A – not available

9000
8000
7000
6000
K mg kg-1

5000 PC
4000 HW

3000 PN
2000
1000
0
380 400 420
Tem perature

Figure 1. Total nutrients in char produced from pine chips (PC), hardwoods (HW), and peanut hulls (PN) pyrolized with steam at three different
temperatures.

At these low pyrolysis temperatures, the total N concentration in the char was similar to that of the initial feedstock
(Table 1). Total N concentration in the PL and PN char was high at 3.7% and 2%, respectively. The PL char
concentration was higher than that reported for active carbon produced from turkey litter (1.12%) although the N
concentration of the turkey litter feedstock was similar at 3.84%(Lima and Marshall 2005b). The turkey litter active
carbon was produced at 700 oC and activated with steam at 800 oC, which may have volatilized more N. The N
concentrations in PC and HW char were similar to those reported for pinewood (0.11%) and oak board (0.18%) char by
Antal and Gronli (2003).

3
 Although the N concnetration of the PN char is potentially high enough to offer a substantial nitrogen input and the
C:N ratio is relatively low (33, Table 1), the N does not appear to be readily available. Nitrogen mineralization was very
low in incubations (24-days at 25 o C, 55% water filled pore space) of infertile, low C Tifton soils (fine-loamy, siliceous,
thermic Plinthic Kandiudults) amended with PN and PC chars at 11 and 22 Mg ha-1 equivalent rate (Table 2). These PN
and PC chars were produced in a pilot scale pyrolysis unit at 400 oC with steam. Peanut hull char C and N concentrations
were 72.85and 1.90%, respectively, and PC char C and N were 76.99 and 0.17%, respectively. There was no statistical
difference in NH4-N concentrations between the control and char amended soils (p=0.05). There was a trend for higher
NO3-N concentrations in the PN amended soils, but only the PN 11 Mg ha-1 rate was statistically different from the
control.

Table 2. Mean change (final –initial) in ammonium- and nitrate-nitrogen concentrations with standard deviations in Tifton soils amended with peanut
hull (PN) and pine chip (PC) char at 11 and 22 Mg ha –1 and incubated for 24 days. Analysis of variance with mean separation by Tukey-Kramer
Multiple Comparison test. Letters within the same column indicate statistical difference at the p=0.05 level.

Feedstock n Δ NH4-N Δ NO3-N

---------------mg kg-1--------------
PN 11 4 1.49 +/- 0.24 5.53 +/- 0.65b
PN 22 4 0.94 +/- 0.78 5.08 +/- 0.62a
PC 11 4 1.19 +/- 0.36 3.62 +/- 0.51a
PC 22 4 1.44 +/- 0.26 4.41 +/- 0.16a
Control 4 1.37 +/- 0.38 3.26 +/- 1.66

We saw similar indications that N in the high N char was not plant available in the first year of field trials on similar
Tifton soils with irrigated corn (Zea mays, Gaskin et al. 2006). Peanut hull char was incorporated to a depth of 15 cm in
microplots (1.8 x 2.2 m) at rates of 0, 11 and 22 Mg ha-1 in a factorial combination with two rates of nitrogen fertilizer (0
and 213 kg N ha-1) surface applied as ammonium nitrate. Tissue samples at the earleaf stage during the first growing
season only showed a significant effect (p=<0.0001) due to N fertilizer. The mean N tissue concentration in the PN
char/no N fertilizer treatments were similar to the control (1.40%) and had average N concentrations averaging 1.13% for
the PN 11 and 1.52% for the PN 22 treatment. Nitrogen tissue concentrations in the N fertilized treatments averaged
2.91%. At the p=0.05 level, there was a significant effect due to N fertilizer for both grain yield and stover (p= <0.0001;
Figure 2), but no significant effect due to char rate for grain (p=0.7197). There was a significant effect of char rate for
stover (p=0.0241). The interaction between char and fertilizer was only significant for the stover (p=0.0145). These
preliminary data indicate, although some N may be mineralizing, the N in the PN char is not readily available to
microorganisms in the short–term (24-days) and is not highly plant available over a growing season (approximately 4
months).

Grain Stover
18
16
14
12
Mg ha-1

10
8
6
4
2
0
22

11

22

11
PN F

PN F

F
l

l
er

er
ro

ro
+

+
PN

PN
liz

liz

PN

PN
nt

nt
22

11

22

11
rti

rti
Co

Co
PN

PN
Fe

Fe

Treatm ent

Figure 2. Corn grain yield and stover (dry wt basis) from Tifton soil plots amended with peanut hull (PN) biochar. Fertilizer – N fertilizer check,
PN22+F- PN char at 22 Mg ha-1 + N fertilizer, PN11+F- PN char at 11 Mg ha-1 + N fertilizer, PN22- PN char at 22 Mg ha-1, PN11 char at 11 Mg ha-1,
Control- no amendment or N fertilizer.
4
 Although N in the char does not appear to be readily available, the char may serve as a source for other nutrients,
particularly K. Mehlich I K at the end of the first growing season was higher in the PN char amended plots than the
control (Table 3).

Table 3. Mehlich I K in Tifton soil amended with peanut hull (PN) char at 11 and 22 Mg ha-1. Analysis of variance with mean separation by Tukey-
Kramer Mulitple Comparision Test. Letters within rows indicate statistical difference at p=0.05 level.

Depth Control Fertilizer PN 11 PN 11 + F PN 22 PN 22 + F

cm --------------------------------------------------- mg kg-1 ---------------------------------------------------------
0-15 28.3 +/- 8.7a 33.2 +/- 12.6a 53.2 +/- 6.8b 49.2 +/- 9.9b 74.1 +/- 11.0c 65.9 +/- 14.3c
15-30 21.3 +/- 4.4a 24.0 +/- 8.4a 43.2 +/- 4.1b 38.4 +/- 7.4b 68.2 +/- 11.3c 51.0 +/- 7.0c

3.2 Feedstock and Pyrolysis Condition Effects on Cation Exchange Capacity

The PN, PC, and HW chars (Table 1) were analyzed for cation exchange capacity (CEC) using a modified Na-acetate
/ethanol/NH4-acetate compulsory replacement method (Sumner and Miller 1996) with sodium analyzed by atomic
absorption spectrophometry. Due to interference of the char ash, chars were leached with deionized water before analysis
to remove soluble salts. In this preliminary study, there was a trend for higher CEC at 400 oC (Table 4). The PN char had
the highest CEC perhaps due to its higher initial mineral concentrations.

Table 4. Mean cation exchange capacity and standard deviation in peanut hull(PN), pine chip (PC) and hardwood (HW) chars produced at three
pyrolysis temperatures with steam.

Temperature
Feedstock 380° C 400° C 420° C
---------------------------------cmol kg-1------------------------------------
PN 36.7+/-0.76 (n=4) 44.0+/-0.35 (n=2) 28.0+/-5.26 (n=4)
PC 18.6+/-1.34 (n=3) 27.0+/-0.60 (n=2) 16.5+/-2.42 (n=4)
HW 22.6+/-0.04 (n=2) 22.9+/-3.21 (n=2) 14.1+/-0.34 (n=2)

3.3 Feedstock and Pyrolysis Condition Effects on Sorption Properties

Subsamples of the PC and PN chars (Table 1) pyrolyzed at 420oC were ground to <420 µm and washed with deionized
water to remove soluble salts and air-dried. Chars were then added to Tifton soils at the rate of 0.05 g char g-1 soil and
phosphorus sorption isotherms were determined using batch techniques. Soil-char mixtures were equilibrated with five
concentrations of P (0, 5, 20, 50, and 100 mg P L-1) in 0.01M CaCl2 matrix. The capacity and intensity of sorption by soil
varied with the type of char added to soil. The amount of P sorbed was highest in soil amended with PN char, while the
lowest P sorption occurred in unamended soil (Figure 3). All systems showed a sharp increase in adsorption at low
equilibrium P concentrations but sorption eventually reached a plateau. This is a characteristic of an L-curve isotherm
where the adsorbate (P) has high affinity for the sorption sites but sorption diminishes regardless of the amount of
adsorbate as surface area decreases (Sposito 1989). Such a relationship suggests a strong interaction between the P and the
exchange surfaces and that the overall sorption was dependent on the properties of both components (Giles et al. 1960;
McBride 1994).

5
 160

140

120
P adsorbed (mg kg-1)

100

80

60
Soil + PC
40
Soil + PN
20
Soil alone
0
0 20 40 60 80 100 120

P concentration at equilibrium (mg L-1)

Figure 3. Phosphorus adsorption isotherm form a Tifton soil amended with pine chips (PC), and peanut hull (PN) chars pyrolyzed at 420 oC with
steam at 0.05 g char g-1 soil .

4. SUMMARY
Soils in the southeastern United States are typically have low C, CEC, and base saturation. Studies of Terra Preta soils
show charcoal has an important influence on these soils productivity and reviews of the activated carbon literature
illuminate some of the physical and chemical mechanisms that could influence soil productivity with char addition.
Pyrolysis chars may have the potential to supply nutrients, sorb cations and anions. The literature and our data indicate
pyrolysis conditions and the feedstock have considerable effects on char characteristics. Our studies on feedstocks
commonly available in the southeastern United States indicate had CECs that potentially increase the ability of low C
loamy sands to retain nutrients. Some chars also have the potential to increase P sorption. It is unknown at this point if
there would be subsequent desorption of P by char and what affect this may have on crops.
Preliminary studies indicate that N from chars with a relatively high N content such as peanut hulls was not readily
bioavailable during the first year of cropping, but some chars contain mineral nutrients such as K that are available to
crops. Our preliminary work indicates char addition may have potential agricultural benefits, but a better understanding
of how char from various feedstocks and produced under different pyrolysis conditions changes soil processes and crop
response is needed.

5. REFERENCES
Alaya, M.N., B.S. Girgi, and W.E. Mourad. (2000). Activated carbon from some agricultural wastes under action of one-
step steam pyrolysis. Journal of Porous Materials 7: 509-517.
Antal, M.J. and M. Gronli. (2003). The art, science, and technology of charcoal production. Industrial and Engineering
Chemistry Research 42:1619-1640.
Asada, T., Ishihara, S., Yamame, T., Toba, A., Yamada, A., and Oikawa, K. 2002. Science of bamboo charcoal: study on
carbonizing temperature of bamboo charcoal and removal capability of harmful gases. Journal of Health Science
48[6], 473-479.
Bansal, R.C., J. Donnet, F. Stoeckli. 1988. Active Carbon. Marcel Dekker, Inc. New York. 482 pp.
Barkauskas, J. (2002). Functional groups on the surface of activated carbons. Part A. Investigation by means of proton
affinity distribution. Chemine Technologia, 24 (3).
Benaddi, H., T. J. Bandosz, J. Jagiello, J. A. Schwarz, J. N. Rouzaud, D. Legras, and F. Beguin. 2000. Surface
functionality and porosity of activated carbons obtained from chemical activation of wood. Carbon 38(5):669-674.
Boehm, H.P. (1994). Some aspects of the surface chemistry of carbon balcks and other carbons. Carbon, 32 (5), pp. 759-
769.
Gaskin, J., L. Morris, D. Lee, R. Adolphson, K. Harris, and K.C. Das. (2006). Effect of pyrolysis char on corn growth and
loamy sand soil characteristics. Abstracts of American Society of Agronomy, Crop Science Society of America, and
Soil Science Society of America International Annual Meetings. Nov. 12-16, 2006. Indianapolis, IN.
6
Georgia Forestry Commission. (2007). Georgia Facts. http://www.gfagrow.org/facts.asp
Giles, C.H., MacEwan, T.H., Nakhwa, S.N., and Smith, D. 1960. Studies in adsorption. isotherms and its use in
diagnosis of adsorption mechanisms and in measurements of specific surface areas of solids. J. Chem.
Soc., London, 3973-3993.
Glaser, B., J. Lehmann, and W. Zech. (2002). Ameliorating physical and chemical properties of highly weathered soils in
the tropics with charcoal – a review. Biology and Fertility of Soils 35:219-230
Gregova, K., N. Petrov, and S. Eser. (1994). Adsorption properties and microstructure of activate carbons produced from
agricultural by-products by steam pyrolysis. Carbon 32(4):693-702.
Guo, Y. and D. A. Rockstraw. 2007. Physicochemical properties of carbons prepared from pecan shell by phosphoric acid
activation. Bioresource Technology, 98 (8), 1513-1521
Harris, K., J.W. Gaskin, L.S. Sonon, and K.C. Das. (2006). Characterization of pyrolysis char for use as an agricultural
soil amendment. American Society of Agronomy, Crop Science Society of America, and Soil Science Society of
America International Annual Meetings. Nov. 12-16, 2006. Indianapolis, IN.
Iswaran, V., K.S. Jauhri, and A. Sen. (1980). Effect of charcoal, coal and peat on the yield of moong, soybean and pea.
Soil Biol. Biochem. 12:191-192.
Iyobe, T., T. Asada, K. Kawata, and K. Oikawa. (2004). Comparison of removal efficiencies for ammonia and amine
gases between woody charcoal and activated carbon. Journal of Health Science 50[2], 148-153.
Liang, B., J. Lehman, D. Solomon, J. Kinyangi, J. Grossman, B. O’Neill, J.O. Skjemstad, J. Thies, F.J. Luizao, J.
Peterson, and E.G. Neves. (2006). Balck carbon increases cation exchange capacity on soils. Soil Sci. Soc. Am. J.
70:1719-1730.
Lehman, J. and M. Rondon. (2006). Bio-char soil management on highly weathered soils in the humid tropics. In:
Uphoff, N., A.S. Ball, E. Fernandes, H. Herren, O.Husson, M.Lang, C. Palm, J. Pretty, P. Sanchez, N. Sanginga,
and J. Theis (eds). Biological Approaches to Sustainable Soil Systems. CRC Taylor and Francis, Boca Raton, FL.
Lehman, J. J. Pereira da Silva, Jr., C. Steiner, T. Nehls, W. Zech, and B Glaser. (2003). Nutrient availability and leachng
in an archealogical Ahtrosol and a Ferrasol of the Central Amazon basin: fertilizer, manure and charcoal
amendments. Plant and Soil 249:343-357.
Lima, I. and W.E. Marshall. 2005a. Adsorption of selected environmentally important metals by poultry manure-based
granular activated carbons. Journal of Chem Technol Biotechnol 80:1054-1061.
Lima, I and W.E. Marshall. 2005. Utilization of turkey manure as granular activated carbon: physical, chemical and
adsorptive properties. Waste Management 25:726-732.
McBride, M.B. 1994. Environmental Chemistry of Soils. Oxford University Press, 406 pp.
Oguntunde, P.G., M. Fosu, A.E. Ajayi, N. van de Giesen. (2004) Effects of charcoal production on maize yields, chemical
properties and texture of soil. Biol. Fertil. Soils 39:295-299.
Perkins, H.,F. (1987). Characterization Data for Selected Georgia Soils. The Georgia Agricultural Experiments Stations,
College of Agriculture, The University of Georgia. Athens, GA. Special Publication 43.
Sonon, L. K. Harris, J. Gaskin, and K Das. (2006). Phosphorus sorption characteristics of Tifton soil amended with
pyrolysis-derived chars. Abstracts of American Society of Agronomy, Crop Science Society of America, and Soil
Science Society of America International Annual Meetings. Nov. 12-16, 2006. Indianapolis, IN.
Sposito, G. (1989). The Chemistry of Soils. Oxford University Press. NY, USA.
Stoeckli, F., A. Guillot, and A.M. Slasli. (2004). Specific and non-specific interactions between ammonia and activated
carbons. Carbon 42 (8-9): 1619-1624.
Strelko, V., D.J. Malik, and M. Streat. (2002). Characterisation of the surface of oxidized carbon adsorbents. Carbon
40(1):95-104.
Steiner, C., W.G. Teixeira, J. Lehman, T. Nehls, J.L. Vasconcelos de Macedo, W. E. H. Blum, W. Zech. (2007). Lang
term effects of manure, charcoal and mineral fertilization on crop production and fertility on a highly weathered
Central Amazonian upland soil. Plant Soil 291(1-2):275-290.
Steiner, C. W.G. Teixeira, J. Lehman, and W. Zech. (2004). Microbial response to charcoal amendments of highly
weathered soils and Amazonian dark earths in Central Amazonia – Preliminary results. In: B. Glaser and W.I.
Woods (eds). Amazonian Dark Earths: Explorations in Space and Time. Springer-Verlag, New York, NY. pp. 195-
212.
Sumner, M.E. and W.P. Miller. (1996). Cation exchange capacity and exchange coefficients. In: Methods of Soil
Analysis. Part 3. Chemical Methods. Soil Sci. Soc. Am. and Am. Soc. Agron. SSSA Book Series No. 5.
Topoliantz, S., J-F. Ponge, and S. Ballof. (2005). Manioc peel and charcoal: a potential organic amendment for
sustainable soil fertility in the tropics. Biol. Fertil. Soils 41:15-21.
USDA. (2002). Census of Agriculture. 29 May 2007. <http://www.nass.gov/Data_and_Statistics/Quick_Stats/>
7
Distributed Hydrogen Production with Profitable Carbon Sequestration: A Novel Integrated Sustainable System for Clean
Fossil Fuel Emissions and a Bridge to the New Hydrogen Economy and Global Socio-Economic Stability

Danny M. Day, Eprida, Inc., 6300 Powers Ferry, Suite 307, Atlanta, Georgia
danny.day AT eprida.com, 404-228-8687
Robert J. Evans, National Renewable Energy Laboratory, Golden, Co
James W. Lee, Oak Ridge National Laboratory, Oak Ridge, TN
Introduction and Abstract

Carbon dioxide (CO2) emissions from fossil fuel combustion directly contribute to ris ing atmospheric CO2 levels, which in
turn are likely linked to global climate change. The need for efficient and economical technologies to rapidly sequester point
source production of carbon dioxide has become both an urgent and widespread technical need. The significant adoption of
any mitigation technology requires a measurable return to end-users. An example is coupling enhanced oil recovery (EOR)
with deep well injection of CO2, though to be economically viable, the carbon dioxide source must be co-located with the oil
fields. We propose an integrated sequestration approach in which agricultural waste products are used to produce hydrogen, a
renewable fuel, and a carbon sequestering soil amendment (char) as a valuable co-product. The dual function char,
derivitized with ammonium bicarbonate (2NH4 HCO3 ) (“ABC”) obtained from treating power plant CO2 emissions, acts both
as an enriched carbon, organic slow release fertilizer and a long-lived carbon storage medium in soils or ECOSS™.

This project had the following objectives:

?To verify a hydrogen co-product could be produced that would offer sufficient value for high volume application
?Test a simply production process that would allow the co-product to be produced from the exhaust of fossil fuel
combustion,
?Analyze the material for characteristics needed of a large volume co-product.

The ammonium bicarbonate technology, developed through the collaboration of Oak Ridge National Laboratory (ORNL), the
National Renewable Energy Laboratory (NREL) and Eprida Scientific Carbons, Inc. (E-SCI), operates at ambient temperature
and pressure and does not require carbon dioxide separation or energy intensive compression. Chars generated from
agricultural waste pyrolysis have been derivitized with the ammonium bicarbonate. This research provides results that point
toward the utilization as a time-release fertilizer while concurrently sequestering stable charcoal in soils and producing an
excess of hydrogen. This hydrogen manufacturing and sequestration strategy utilizes the largest existing market for hydrogen
(i.e. the production of fertilizer) and leverages the existing farm fertilizer infrastructure to restore soil carbon lost by erosion
and extensive tillage. An ancillary benefit of this process is the accrual of carbon credits from capturing power plant
emissions, producing a long-lived carbon soil amendment, and enhancing plant growth. If this integrated strategy ultimately
proves successful, then the agricultural sector can play an inte gral role in developing the hydrogen economy and restore soil
carbon, sequestering vast amounts of carbon, while increasing available nitrogen and sustained natural sequestration will
occur through enhanced plant growth. Implementation could allow hydrogen production to work synergistically with fossil
fuel emission reductions. A global economic stimulus is possible through profitable sequestration, increased farm
productivity, local energy production, rural income opportunities, small business development and offer hope for a positive
and sustainable future.

1
Step 1. Hydrogen Production Leveraging Photosynthesis
Extract Hydrogen From Organic Matter Catalytic Steam
Reforming
60% H2
20% CO2
7% CO
3% CH4
100-Hour Field Demonstration of
Hydrogen From Biomass Catalytic Steam
Reforming (August 2002)

Plus 20% (by weight) Charcoal

But why produce charcoal?

2
Step 2: Evaluation of Charcoal as a Sequestration Media and
Carrier for Plant Nutrients
A Valuable Co-Product?
We began to investigate the use of the material as a soil amendment and nutrient The
carrier after employees mentioned that a mound of char, used to supply char for
start-up operations was covered in vegetation and more specifically turnips.
missing
Someone had tossed some turnip seeds, on the two year old, chest high, char pile. turnips
It was comprised only of char with no soil, yet on plants completely covered the
mound. The plants appeared healthy with roots that enveloped each char particle.
The turnips, unfortunately could not be inspected as they already had been eaten,
but it was reported they were “Good!”.

Charcoal Background
? Forest fires have been sequestering carbon in the
soil in as charcoal for billions of years
? The half-life of charcoal in the soil is measured in
1000’s of years (Skjemstad)
? Charcoal even in weathering environments can be
found as old as 11,000 years old (Gavin)

Charcoal is a natural part of all soil carbon content

Carbon in the Soil

?Char is a pyrogenic carbon, often lumped under the classification of black carbon
? Black carbon is a term widely used for soot, an amorphous residue of combustion and a
contributor to global warming. Char is not soot.
? Char found in soils from forest and range fires has a carbon framework remaining after
the pyrolysis of volatile organics.
? Charcoal has proven itself with over 2000 years of testing as a soil amendment in terra
preta soils.

The Terra Preta Soil Experiment,

2000 Years Old
Terra Preta refers to black high carbon (9%) earth-like anthropogenic soil with
enhanced fertility due to high levels of soil organic matter (SOM) and nutrients
such as nitrogen, phosphorus, potassium, and calcium. Terra Preta soils occur
in small patches averaging 20 ha. These man-made soils are found in the
Brazilian Amazon basin, also in Western Africa and in the savannas of South
Africa. C14 dating the sites back to between 800 BC and 500 AD. Terra Preta
soils are very popular with the local farmers and are used especially to produce
cash crops such as papaya and mango, which grow about three times as rapid
as on surrounding infertile soils.
(Map reprinted by permission: Steiner, 2002)

3
 Current Research

The images above were provided for this poster by Christoph
Steiner, who has been recreating Terra Preta soils in Brazil since
1999.
•Amount of applied organic matter (25% increase of Corg in
0-10 cm
• Increased the soil C content ~ 0.75%
•Applied Charcoal 11 t / ha
•Mineral fertilizer: N (30), P (35), K (50), lime (2100 kg/ha)
International Workshop on Anthropogenic Terra Preta Soils,
(July 2002 Brazil)
Pyrogenic Carbon Benefits
• Terra Preta soils contain 15-60 Mg/ha C in 0-0.3m but 1-3Mg/ha may be
sufficient to enhance plant growth (GLASER et al.)
• Surface oxidation increased cation exchange capacity (GLASER)
• Char decreased leaching significantly (LEHMANN)
• Char traps nutrients and supports microbial growth (Pietikainen)
• Char increased available water capacity by more than 18% than
surrounding soils (GLASER)
• Char experiments have shown up to 266% more biomass growth
(STEINER) and 324% (Kishimoto and Sugiura)

Characteristics of an Optimized Pyrogenic Fertilizer

1. A source for microbial Leaching Examination of Different Chars
energy
10
2. Low leaching rates 9.5 900 C
9
3. Internal pore structure 600 C
pH

8.5
for deposition 8 450 C
7.5 400 C
4. Good cation exchange 7
2

10

12
il
ita
Int
pH

100ml Rinse - Char Sample 20.0g

Intra-particle volatile fatty acids deposits (Runkle et al.) occurring inside exothermic zone of pyrolysis are an
excellent microbial energy source (Westjohn et al.) for processing nitrogen compounds. We speculated that near the
dew point temperature a suitable material might require no outside fuel.
Leaching experiments were conducted of materials from different temperatures. Crushed and sieved (US #30
mesh), 20g samples was soaked for 5 minutes in 48% NH 4 NO 3 solution and allowed to dry for 24 hours. Samples
were leached with 100 ml rinses ( 8.0 pH). The 400C sample displayed very low leaching rates and was chosen as a
starting material.

4
 Step
Step 3: 3: Process
Process MapMap
An additional benefit of the process is its ability
to combine with SOx and NOx from fossil fuel
exhaust. Both of the molecules are scrubbed
from the exhaust to form value added nutrients.

Process Flow : H2 Production w/ECOSS (Patent Pending)

Forest Residue / Energy Crops

H2 + CO2 CO2
Pressure Swing
Adsorption
H2 (73%) Use/Sell

H2 Purifier/Dryer N2
Steam (27%)
(or alternate H2 System)
Steam Reformer

Compressor

Pyrolysis Pyrolysis Pyrolysis

Reactor Reactor Reactor
Catalytic
#1 #2 (Opt) #3 (Opt) Heat Converter
Exchanger

Pyrolysis
Off-Gas Recycling Condenser
Water
Char Char Char Pump
Nutrient Mix/PK Clean
Ammonia Exhaust
Char
(Opt. Ammonia Purchased)
Metering Blending
Fluidized
Fossil Fuel Gases w/ CO 2/SO x/NOx Multistage Release
Cyclone ECOSS Fertilizer

5
 Step 4:
Step 4: Bench
Bench
Scale Reactor
50 kg of char was prepared at 400C.
After initial heat up, no additional heat
was added and the pyrolysis unit ran
exothermically. The high temperature
rotary value discharged the char into a
closed container where it cooled. The
granular material was ground and
sieved to a uniform particle range. (20-
30 US Mesh)

The reactor was fed powdered char, ammonia (saturated

with water), and CO2. A variable speed rotor suspended the
particles and as the ammonia and CO2 derivitized
ammonium bicarbonate, they would gain mass and move
down the cyclone until reaching the discharge cyclone. The
speed of the rotor controlled average residency time. The
sand like material formed within 5-15 minutes and the
larger granules (Nitrogen >10%) between 15-30 minutes.

Step 5: The exterior images show the visible build

up on the outside of the particles. After
Analysis crushing a particle, the formation of the
ammonium bicarbonate inside the fractures
and interior cavities can be clearly identified.
Exterior

Formation of ABC in
Fractures

Char Slow Release

Mechanisms
Exterior Buildup as Expected

Volcano like Sizable Interior Cavities

Sand like Structures
around pores

Granular Interior View 422x

6
The SEM to the left is of the sand like material
and was taken of the crushed interior. The
charred carbon framework of the plant cell Interior Image (Sand Like - 2000X)
structure is visible, appearing like plastic. The
volatile organics inside this char provide the
needed energy source for microbial action. The
ammonium bicarbonate appears as cotton-like
Charred
fibers.
Plant
The image below shows ammonium Framework
bicarbonate has filled the interior of the
large granular particles.

ABC Deposits

Granular Interior – 1000X

Step 6: Potential Impacts Evaluation

Carbon Dioxide per Million BTU

Bituminous

Diesel

Gasoline
Fuel

Propane
CO2 kg/MBTU
Liquified Petroleum Gases (LPG)

Natural Gas (Pipeline)

H2 & ECOSS

-100 -50 0 50 100

Current energy use ~400Ej/yr (Lysen) and CO2 is increasing by 6.1 Gt/yr
(IPCC). Each 1.0 MBTU H2/ECOSS represents 91kg (as measured, not
calculated) of sequestered CO2, then 6.1Gt/91kg equals 0.07Ej or 0.01.8% of
the current world consumption of energy.

7
 Conclusions
Material Balance and Production Limits (Energy is not the limiting factor) At theoretical
maximum H2 –CO2 conversion there would only be enough CO2 to convert 61% of H2 to
ABC and since our target nitrogen content for the pyrogenic carbon is 10%, (requiring 45%
carbon by weight), our limit becomes the 20% carbon char (wt. 12) vs the 56% of ABC
(mol.wt. 79). The limit is therefore the carbon char as a carrier utilizing only 27% of avail-
able hydrogen but sequestering 91kg of carbon dioxide (as measured experimentally) per
million BTU of hydrogen utilized for energy. In addition, there is more than 91kg when the
carbon sequestered in the form of additional plant growth and CO2 equivalents from reduced
greenhouse gas emissions from lower power plant and fertilizer NOx release.

? Hydrogen and carbon sequestration can be economically combined.

? Production of a valuable sequestering co-product during hydrogen production has a

potentially large volume application
? The material displays characteristics which will reduce nutrient leaching and loss

? Production of a nutrient inside carbon pores for physical slow release mechanism possi-
bly enhancing plant uptake and reducing fertilizer GHG emissions.
? Physical micro pore structure offers safe haven for enhanced microbial activity and in-

creased soil fertility

? Intra-particle deposition of volatile fatty acids offer microbial energy source and enhance-
ment of nitrogen compound processing.

? Increased cation exchange and water holding capacity provide better plant-soil efficien-
cies.
? Provides a stable sequestration method where existing fertilizer business infrastructure

can profit
? Offers forestry and agriculture a method to enhance carbon sequestration and fiber/crop
yields
Danny Day
danny.day AT eprida.com Eprida
404-228-8687 6300 Powers Ferry Rd, Suite 307
http://www.eprida.com 8 Atlanta, GA 30339
 The Potential for Biofertilisers
The growth in agricultural production during the last three decades has been accom-
panied by a sharp increase in the use of chemical fertilisers, causing serious concern
(Marothia, 1997). Foremost among these concerns is the effect of excessive fertiliser
(especially nitrogenous fertilisers) on the quality of soil and ground water.8

Biofertilisers are considered to be an important alternative source of plant nutrition.

They are biologically active products, including bacteria, algae or fungi, with the ability
to provide plants with nutrients. Most biofertilisers belong to one of two categories:
nitrogen fixing and phosphate solubilising. Nitrogen fixing biofertilisers fix atmos-
pheric nitrogen into forms which are readily useable by plants. These include rhizo-
bium, azatobacter, azospirillum, blue green algae (BGA) and azolla. While rhizobium
requires symbiotic association with the root nodules of legumes to fix nitrogen, others
can fix nitrogen independently. Phosphate solubilising micro-organisms (PSM) secrete
organic acids which enhance the uptake of phosphorus by plants by dissolving rock
phosphate and tricalcium phosphates. PSMs are particularly valuable as they are not
crop specific and can benefit all crops (Table 2).

Table 2. Major biofertilisers and target crops

Biofertiliser Target Crop

Rhizobium Leguminous crops
(Pulses, oilseeds, fodder)
A zatobacter W heat, rice, vegetables
A zospirillum rice, sugarcane
Blue green algae (BG A) rice
A zolla rice
Phosphate solubilising microorganisms (PSMs) all

Production of biofertilisers in India

The idea of using micro-organisms to improve land productivity has been around in
India for at least 70 years, but it was only in the 1990s that large scale production of
various biofertilisers commenced. Presently, a number of agricultural universities, state
agricultural departments and commercial enterprises produce various biofertilisers.

The promotion of biofertilisers is mainly carried out by the National Biofertiliser Devel-
opment Centre (Ghaziabad), which was set up in 1987. The main objectives of the
National Centre are to:

8 Water containing excess of nitrates can affect the blood’s ability to transport oxygen, with serious health implica-
tions (WHO, 1963).

8 G ATEKEEPER SERIES N O .SA93

• produce and market biofertilisers of required quality;
• isolate and maintain biofertiliser strains suitable to various agro-climatic regions;
• train agricultural extension workers;
• promote biofertilisers through field demonstrations;
• prepare quality parameters;
• test samples of biofertilisers produced by others;
• provide technical and financial assistance to units producing biofertilisers.

The National Centre has the capacity to produce 375 tons of biofertilisers per year. In
addition to this, 58 commercial production units have been set up with government
support. India’s total production in 1998-99 was reported to be 16,000 tons.9 Rhizo-
bium accounts for the largest proportion (40%) of the total production in India. This
is followed by azatobacter. With the increase in the price of phosphate fertilisers, the
potential for the use of PSM has also increased.

Effectiveness of biofertilisers
A considerable amount of research has been done to establish the effectiveness of biofer-
tilisers on various crops, in different agro-climatic regions. Most agricultural universi-
ties, the ICAR and the National Biofertiliser Development Centre have carried out a
number of field trials to document the effectiveness of these micro-organisms. These
programmes show that the use of biofertilisers can have a significant effect on the yield
of most crops. However, their effectiveness is found to vary greatly, depending largely
on soil condition, temperature and farming practices. As an example, Table 3 shows the
effect of azatobacter on yield.

Table 3. Effect of azatobacter on crop yield

Crop Increase in yield over Crop Increase in yield over

yields obtained with yields obtained with
chemical fertilisers (%) chemical fertilisers (%)
Food grains O ther
W heat 8-10 Potato 13
Rice 5 Carrot 16
M aize 15-20 Cauliflo w er 40
Sorghum 15-20 Tomato 2-24
Cotton 7-27
Sugarcane 9-24
Source: Das, 1991

9 Dr. T. Singh, pers. comm., Director, National Biofertiliser Development Centre.

G ATEKEEPER SERIES N O .SA93 9

 The Case Study:
Karnal and Bhiw ani Districts, Haryana
A field study was initiated in two districts in Haryana to try to understand why biofer-
tilisers and biopesticides were not being adopted on a wide scale. The study focused
on two districts: Karnal and Bhiwani. While Karnal represents intensive agriculture
with a high degree of irrigation, Bhiwani represents dryland farming, with low level
irrigation. The potential for biofertilisers and pesticides is described below, followed
by the results of our interviews with farmers to establish why biofertilisers and biopes-
ticdes have not been taken up more widely.

The Potential for Biopesticides and Biofertilisers in Haryana

The problems caused by excessive pesticide use are particularly serious in rice and
cotton in Haryana, both important crops in our study districts. The American boll-
worm, jassid and white fly in cotton; and stem borer in rice; have developed resistance
to chemicals in many parts of Haryana, including parts of the districts studied. In this
situation there is great potential for biopesticides.

The potential of biofertilisers is evidenced by the fact that about 90% of Haryana’s soil
is deficient in nitrogen, indicating severe nutrient deficiency (Dahiya et al., 1993). The
dryland districts such as Bhiwani, Mohindergarh, Sirsa and Faridabad are particularly
low in nitrogen, and soils are short of organic matter due to poor vegetative cover, high
temperatures and the light texture of soil. Biofertilisers could play a role in providing
the much needed nutrients and improving soil conditions in these dryland areas, which
account for about 28% of the state’s total cultivable area.

Biofertilisers can also reduce the intensity of chemical fertiliser consumption, especially
in irrigated areas. With the increase in cropping intensity (see below), the use of chem-
ical fertilisers has increased significantly in these areas. The use of organic manure, on
the other hand, has not increased. As a result, many parts of Haryana face deteriorat-
ing soil conditions and increasing ground water contamination.

The suitability of biofertilisers for various crops grown in Haryana has been shown
through demonstrations conducted by the Hissar Centre of the National Biofertiliser
Development Centre (NBDC). Azatobacter and azospirillum, which can be used for a
wide range of crops, are estimated to have particularly large potential. For example,
NBDC experiments showed an increase in yield between 3 and 25% with the applica-
tion of azotobacter in cotton. Similarly, in the case of wheat, use of azotobacter resulted
in yield increases between 2 and 20%.10

10 National project on use and development of biofertilisers. Biofertiliser News, 1(2), December 1993.

10 G ATEKEEPER SERIES N O .SA93

The potential for the use of rhizobium is largely confined to the dryland areas of
Haryana, where pulses are commonly grown, and where the soils show poor to moder-
ate nodulation.11

Karnal District
In Karnal the main nutrient and pest problems relate to the district’s high cropping
intensity. Wheat-rice rotation has been common for some time now, and cropping inten-
sity has recently increased further with the introduction of short duration rice varieties
such as Govinda (90 days). Planting these varieties allows farmers to take two rice crops
between April and October. The first paddy crop is planted in April-May, as soon as
wheat has been harvested. This rice crop is ready for harvest by mid July. This is
followed immediately by the transplanting of fresh paddy seedlings. The second rice
crop is harvested by the end of October, to be followed by wheat, which is sown by
early November.

This very high intensity of cropping has worsened both the soil quality and pest problem
in parts of Karnal. It is extremely damaging to the soil condition, as large amounts of
nutrients are used continuously without replenishment. As a new crop is planted as
soon as the older crop is harvested (sometimes both activities are done simultaneously),
there is no time for proper land preparation and for using organic manure. As the use
of chemical fertilisers has increased to provide the required nutrients, soil and water
conditions have deteriorated.

The problem of pests and pesticide use in Karnal is largely confined to rice. Growing
two rice crops without a break is one factor. The continuation of rice plants in the same
field, and the high degree of moisture, enable pests to multiply without a break, leading
to particularly intense pest attacks in the second crop. Another reason is the popular-
ity of certain basmati varieties which are highly susceptible to pests and diseases. This
is particularly serious in one of the most popular of these varieties, called duplicate
basmati. This variety was introduced about five years ago from West UP, and became
very popular because of its high yields,12 and totally replaced the desi (traditional)
basmati variety in many areas. After performing well for three years, the variety began
to be affected by pests (stem borer, leaf folder and white back plant hopper) and diseases
(sheath blight, blast and bacterial leaf blight) two years ago. The attack was particu-
larly severe last year, forcing many farmers to stop planting basmati in general, and
duplicate basmati in particular. In one of the villages we studied, Kuuchhpura, farmers
made up to 15 pesticide applications last year but could not save the crop. This year,
very few farmers have planted basmati in the village.

11 The assimilation of atmospheric nitrogen by the roots of pulse (and other leguminous) crops is carried out
by the nodules formed in the roots of these plants. The low degree of nodulation suggests that their ability to
assimilate atmospheric nitrogen is low and that they could benefit from the use of rhizobium.
12 Compared to the average yield of 10 quintal/acre from desi basmati, the yields of duplicate basmati were
about 20 quintal/acre.

G ATEKEEPER SERIES N O .SA93 11

The problem of pesticide resistance in stem borer is now common in Karnal, largely
caused by the indiscriminate and excessive use of pesticides. Another important reason
relates to the fact that farmers in Karnal do not spray their fields themselves. This is
done by professional workers who are sent by the shops selling pesticides. As these
workers are paid by the area covered, they do a rushed job. The spraying is not uniform:
while some parts of the fields get very little, others get excessive pesticides. Secondly,
in order to save time they use about one third of the water prescribed to make the pesti-
cide solution. The non-uniform spray of highly concentrated pesticide solution is
reported to be a major reason for the development of resistance in rice pests in many
parts of the district.13

Bhiw ani District

In Bhiwani, the focus of the study is on cotton and gram, which, along with wheat and
mustard are the main crops in the area. The use of chemical fertilisers is comparatively
low in Bhiwani, being used only on wheat and cotton, and not at all on gram and
mustard. But, as in Karnal, use of organic manure is not common and the condition of
the soil is poor. Three biofertilisers have potential for Bhiwani: rhizobium for gram and
azatobacter and PSM for wheat. The potential of rhizobium is reported to be particu-
larly high in Bhiwani because the level of nodulation is very low.

The District Agriculture Department is responsible for the sale of rhizobium in Bhiwani.
Being a major gram growing district, it was allocated the largest amount of rhizobium
last year and its sale to farmers is subsidised at the rate of 50%.

The main pest problem in Bhiwani concerns cotton, which is attacked by American,
pink and spotted bollworm, white fly and jassid. These pests have become resistant to
most pesticides. According to a study of pesticide use in Bhiwani in 1998, about 70%
of farmers reported that they were unable to control pests with pesticides (Saini and
Jaglan, 1998). Some of these farmers had used up to 11 pesticide applications.

The pest problem in Bhiwani is closely linked with the spread of irrigation facilities.
Large parts of Bhiwani district are irrigated by the Indira Gandhi Canal, which suffers
from large scale seepage. This has led to a rise in the groundwater level from 50-60 feet
below the surface in the past to 5-10 feet at present in many areas. As a result, water
logging and high humidity are serious problems in many areas of the district. Apart
from causing direct damage, this has also created favourable conditions for the growth
of pests. Consequently, the problem of pests has become extremely serious in the district
during the last five years.

The pest problem caused the area under cotton to decline in the second half of the
1990s from 57, 000 hectares in 1996-97 to 51,000 hectares in 1999-2000.14 As in rice

13 Information from the District Agricultural Officers.

14 Information provided by the State Agricultural Department, Bhiwani.

12 G ATEKEEPER SERIES N O .SA93

in Karnal, the pest problem in cotton in Bhiwani is closely related to the susceptibility
of the varieties being planted. The cultivation of cotton in this area is comparatively
new: it was introduced in the 1970s. Most of the varieties introduced since then are
long staple, American types. Although they fetch high prices, they are highly suscepti-
ble to various cotton pests. The problem of pests and the excessive use of pesticides in
Bhiwani is mostly confined to the American varieties. Desi varieties, which are compar-
atively resistant to pests and diseases, require less use of pesticides.

We observed a shift back in favour of desi cotton in one of the divisions of Bhiwani
following severe losses to pests to the American varieties. Whilst many more farmers
would prefer to shift to desi cotton varieties, as they provide stable yields, their whole-
sale prices are too low. In addition it appears that the state agriculture department
prefers to promote long staple American varieties, as they have large domestic and
export markets.

Farmers’ Perspectives
We selected 11 villages from the two districts, six villages from Karnal and five from
Bhiwani, in which to interview farmers to establish their awareness and use of biofer-
tilisers and biopesticides. A total of 74 farmers from these villages were interviewed, and
were mainly selected because they had attended one of the demonstration programmes
carried out by government agencies to promote IPM, biopesticides and biofertilisers
(58 of the farmers, or about 80% of the sample). However, some farmers who had not
been to the programmes were also interviewed (16 farmers). All the farmers were men;
only men had participated in the demonstration programmes, and we were told that
men take the decisions about the types of fertilisers and pesticides to be used.

The farmers included in the study represent the range of small, medium and large
farmers. While 29% of the farmers have less than 5 acres of land, 46% have between
5 and 20 acres. 25% of the farmers have landholdings larger than 20 acres. In terms
of education, about one third of the farmers have studied up to class 10 or more. A
semi-structured questionnaire was used for interviews, which were conducted in
farmers’ homes.

Findings: Biopesticides
Although biopesticides and bio-control agents are important components of IPM, the
IPM programmes being conducted by various agencies put very little emphasis on these
agents. None of the farmers we interviewed had ever used biopesticides, and few were
even aware they existed. We found that none of the farmers had even used neem, widely
believed to be commonly used by Indian farmers.

In fact, we found that IPM itself was not being practised by most farmers. While some
of the farmers were aware of IPM practices, such as the need for monitoring and

G ATEKEEPER SERIES N O .SA93 13

augmenting natural enemies of pests, very few farmers have adopted these practices.
None of the farmers practised IPM fully, but about 15% practised partial IPM. In most
cases this meant delaying the first spray of pesticides until damage by pests becomes
evident. The situation is particularly bad in Bhiwani, where 85% of farmers reported
not using IPM at all and only 9% reported using some aspects of IPM. Compared to
this, 20% of farmers reported using limited IPM in Karnal. This is despite the fact that
most of the farmers included in the study participated in the IPM demonstration
programmes conducted by the Central IPM.

W hy is IPM not practised?

We found four important reasons for the low acceptance of IPM:

1. Lack of awareness. This reason was found to be particularly important in Bhiwani,

where 77% of farmers were unaware of the concepts of IPM. Compared to this, fewer
than 3% of farmers in Karnal were unaware of IPM. Clearly, there is a big district-
wise difference in the success of IPM demonstration programmes and state agricultural
extension workers in familiarising farmers with IPM. In Karnal these agencies have
been successful in at least making farmers aware of the need to practise IPM.

2. Lack of skills. Almost all the farmers, including all of those who were aware of IPM,
reported that they lacked the skills necessary to practise IPM. Their ability to practise
IPM was, for example, severely constrained by the fact that most of them could not differ-
entiate between harmful and beneficial insects. In fact, many farmers thought that all
insects were a potential threat to their crop and had to be destroyed. They were also not
able to work out economic thresholds15 to determine the timings of pesticide application.

3. Lack of faith in IPM. This factor was found to be very important amongst 60% of
farmers in Karnal. Although almost all of them were aware of IPM, they felt that they
did not have sufficient faith in it to reduce the use of chemical pesticides. Many of
them felt that the support from the agricultural department was not adequate for them
to try IPM practices, which were considered risky. As the CIPM personnel do not keep
in touch with the farmers after IPM demonstrations, they felt that they would not get
adequate advice and support if things went wrong. Similarly, the local extension
workers (ADOs) are not sufficiently trained in IPM to instil confidence in the farmers.

The issue of skills and confidence is obviously linked to the intensity of training
provided by the IPM agencies to farmers and extension workers. We found that the
training is very basic and superficial, being conducted for three to four days in a
village, during which 30 villagers and five extension workers are trained. The farmers
felt that the training was not intensive and did not impart sufficient skills for them
to feel competent and confident in following IPM practices.

15 The economic threshold is the level of pest population at which the damage to crop justifies the use of pest
control methods.

14 G ATEKEEPER SERIES N O .SA93

4. IPM practices are difficult and cumbersome. About 70% of the farmers in both
districts who were aware of IPM, felt that the IPM practices were too cumbersome
and time consuming to be used regularly. Both the monitoring of pest populations
and the calculation of economic thresholds were considered by farmers to be imprac-
tical. In Karnal, where cropping intensity is high, the farmers felt that they did not
have time to keep a close watch on their fields to monitor pests and calculate
economic thresholds.

W hose advice is taken?

An important reason for the failure of the IPM programme, and the lack of biopesti-
cide use, is related to the fact that most farmers depend on shopkeepers for advice on
pesticide application. More than 80% of the farmers reported that shopkeepers, dealers
and representatives of pesticide manufacturers were their most important sources of
information about pest control methods (Table 4).

Table 4. Importance of source of advice for pesticide application

O ther Extension Shops/manufacturers/

Farmers w orkers and dealers
agricultural university
Karnal 12 (30) 18 (45) 33 (83)
Bhiw ani 10 (30) 9 (26) 27 (80)
Total 22 (30) 27 (36) 60 (81)
Number of responses=74. Note that some farmers listed more than one source of advice as being
equally important
Note: Figures in parenthesis indicate percentages.

Compared to the manufacturers and sellers of pesticides, agricultural extension workers

play a small role, especially in Bhiwani, where only 26% of farmers reported them to
be important.

The role of shopkeepers and dealers is particularly important because many farmers
(58%) purchase their pesticides on credit. This gives the shopkeepers strong control
over the amount and choice of pesticides used. It also makes it easier for the shop-
keepers to sell spurious pesticides.

Finally, we found that the knowledge of the State Agricultural Department about
biopesticides was extremely limited. This was particularly true of the village level
workers (ADOs), but also the case even in the Central IPM office in Faridabad. Consid-
ering the importance given to the use of biopesticides by national government agen-
cies, the neglect of biopesticides at the state and district level is difficult to understand.
The main emphasis is on the monitoring of pest populations and the use of economic
thresholds, which farmers find too difficult and time consuming. Further, the Haryana

G ATEKEEPER SERIES N O .SA93 15

Agricultural University recommends the use of only two biopesticides (Bt and neem) on
only one vegetable crop. On all other crops, including cotton and rice, the recommen-
dations include only chemical pesticides.

Findings: Biofertilisers
Few (19%) farmers have ever used biofertilisers in the two districts. Their number was
especially small in Karnal, where only three out of 40 farmers had used them. In
Bhiwani about 75% of farmers were not even aware of biofertilisers; the proportion of
such farmers was even higher in Karnal (85%).

The most important reason for this lack of awareness is the fact that agriculture exten-
sion workers do not promote biofertilisers. On the whole, only 15% of farmers had
been told about biofertilisers by the extension workers. The emphasis on biofertilisers
was particularly low in Karnal, where only one farmer out of 40 had heard about them
from extension workers.

The District Agricultural Departments do not have a positive attitude towards biofer-
tilisers. They feel that their quality is poor, and their performance totally unreliable.
Therefore, they are not prepared to risk their reputation and good will with the farmers
by recommending them. The extension services run by the Haryana Agriculture Univer-
sity (krishi vigyan kendra) feel that in areas of wheat/paddy rotation, such as Karnal,
the potential of biofertilisers is low. The farmers in these areas can get the same yield
by using the recommended dose of chemical fertilisers. As these fertilisers are easy to
use, the farmers prefer them. Biofertilisers have to be stored and applied in conditions
which are suitable for the multipication of micro-organisms. This requires special facil-
ities and care, which farmers are often unable to provide. Chemical fertilisers, on the
other hand, can be stored and applied without special care. In fact, the KVK in Karnal
does not recommend the use of biofertilisers at all.

Nevertheless, the extension workers in Bhiwani are required to sell a fixed number of
rhizobium packets. This explains the larger number of farmers who are aware of biofer-
tilisers, and have used them in this district. But we found that a large proportion of the
rhizobium allotted to the district is not sold to the farmers, and is allowed to go to
waste. The ADOs say that the quality and performance is so poor that the farmers are
not interested in buying it. The quota is shown as sold in official records, and the
payment is made by the ADOs from their salary.16 The fact that the rhizobium meant
for sale in various pulse growing districts is not being used by farmers is widely known
to both the State Agricultural Department and the National Biofertiliser Development

16 For example, one of the ADOs in Bhiwani was given a quota of 2000 packets of rhizobium this year. He
could manage to sell only about 200 packets, mostly to his contact farmers. But these have not actually been
used by the farmers. He feels that the contact farmers accept biofertilisers because they want to stay in the good
books of the ADOs, from whom they receive subsidised or free goods, such as seed kits. He will have to pay
to the Agricultural Department Rs. 8,000/- (@Rs. 4.00 a packet).

16 G ATEKEEPER SERIES N O .SA93

Centre. However, every year the practice of fixing quotas and reporting of sale is carried
out, as if rhizobium is actually being used.

Out of 19 farmers who were aware of biofertilisers, 14 had used them, showing that
farmers are prepared to try biofertilisers. However, most of these farmers stopped using
them after one crop - only three were still using them. Two reasons were reported to
be important for the discontinuation of use: lack of availability and poor performance.

Availability
Many farmers who stopped using biofertilisers reported that this was partly because the
supply was extremely unreliable. This was largely because biofertilisers were not being
sold by most shops. While none of the shops stock biofertilisers in Karnal, two shops
in Bhiwani do. The shopkeepers, in turn, say that they do not stock biofertilisers
because sales are poor. One of the Bhiwani shops, for example, has been stocking biofer-
tilisers from National Fertilisers Ltd. for the last four years but has sold only 30 packets.

Q uality
It is clear that the poor quality and performance of biofertilisers present serious prob-
lems. Most studies suggest that the biofertilisers being sold in the market are contam-
inated and have a low count of micro-organisms. It is therefore not surprising that their
performance is poor and uneven. For example, in a survey of rhizobium carried out by
ICRISAT, 90% of samples from India were found to have a rhizobia count lower than
that required for effective performance (Singleton et al., 1996). Incidentally, this
problem exists in most developing countries. For example, in a survey of 12 develop-
ing countries, only 19% of the samples met the standards prevalent in developed coun-
tries (Singleton et al., 1996).

The poor performance of biofertilisers in India is primarily linked to inappropriate

strains and inefficient production technology. Essentially, the production of bacterial
biofertilisers requires the selection of strains appropriate for a particular crop in a given
agro-climate. These strains are mass multiplied by adding bacterial culture to a suit-
able sterilised broth, either using the shake flask method (for small scale production)
or the fermenter method (for large scale production). When an adequate growth of
bacteria is achieved, the solution is mixed with a carrier such as lignite or charcoal and
is packaged.

As agro-climatic conditions and soil characteristics vary widely, a large range of strains
of each biofertiliser needs to be isolated for each area. The problem of identifying suit-
able strains is particularly serious in north India, as many of the strains do not survive
the very hot temperatures prevalent in these areas. Until strains which can tolerate wide
variations in temperature can be identified, the performance of biofertilisers will remain
uneven. The Haryana Agriculture University is reported to be working in this direction
and has developed improved strains for pearl millet, wheat, mustard, potatoes, and
flowering plants. These are, however, yet to be used in large scale production.

G ATEKEEPER SERIES N O .SA93 17

Furthermore, the production of biofertilisers is prone to contamination, which reduces
the effectiveness of micro-organisms. It is, therefore, vitally important that throughout
the process extreme care is taken to maintain sterile conditions. It is also important
that precautions are taken to avoid contamination during the packaging, storage and
application of biofertilisers.

The production technology employed in India is inefficient and is responsible for most
of the contamination common in Indian biofertilisers. Generally, production is under-
taken by the flask method, which is unsuitable for large scale production. Although
some firms use fermenters, they lack the sophisticated controls and monitoring facili-
ties necessary to regulate factors such as pH, temperature and aeration. As a result the
quality of the bacterial broth is often poor and uneven.

Another problem relates to the fact that Indian producers do not sterilise the carriers
used for mixing the bacterial solution. For example, both the producers in Haryana, the
Hissar Biofertiliser Centre and the HAU, use non-sterilised charcoal, as they do not
have facilities to sterilise large amounts of charcoal in a short time. This is an impor-
tant cause of the poor quality and short shelf life of these biofertilisers (Singh et al.,
1999). Although India has ISI (Indian Standards Institution) standards for some of the
biopesticides (rhizobium and azatobacter), they are not enforced. This is reportedly
because the ISI lacks facilities to test biofertilisers.

Conclusions
Biopesticides
The problem of pests, the development of pesticide resistance and the excessive use of
pesticides need to be seen in the totality of the agricultural system. Our study shows that
in Karnal and Bhiwani the problem is linked to the increase in cropping intensity (three
crops in Karnal), the expansion of irrigation facilities (Bhiwani), the release and adop-
tion of susceptible varieties (govinda and basmati rice in Karnal, and American cotton
varieties in Bhiwani), purchase of pesticides on credit (in both the districts) and inap-
propriate agricultural practices (the use of contract labour for pesticide application,
using power spraying machines in Karnal). In the circumstances, mere reliance on pest
control, without correcting the basic problem in the system, will not produce sustain-
able results.

The efforts of various government agencies to popularise integrated pest management

(IPM), and the use of biopesticides have had little impact. IPM departments have very
little knowledge and experience of biopesticides, and most state agricultural universi-
ties, on whose recommendations pest control methods are promoted, do not include the
use of biopesticides in their recommendations. In the absence of active promotion by
the agriculture department, the demand for these products has not developed. It is for
this reason that most private shops and dealers do not stock and sell biopesticides.

18
18 G ATEKEEPER SERIES N O .SA91
.SA93
 Gatekeeper Series no. 93

A Study of
Biopesticides and
Biofertilisers in
Haryana, India

Ghayur Alam

2000
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The Gatekeeper Series produced by IIED’s Sustainable Agriculture and Rural Liveli-
hoods Programme aims to highlight key topics in the field of sustainable agriculture
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Dr Ghayur Alam is Director of the Centre For Technology Studies in New Delhi. His
main areas of interest include technology policy, intellectual property rights,
agricultural biotechnology and the adoption of environmentally suitable technologies
in industry and agriculture. He can be contacted at: Centre For Technology Studies,
U 24/20, DLF III, Gurgaon, Haryana 122002, India. Email: alamcts@ndf.vsnl.net.in
Tel: +91 (0124) 6355942; Fax: +91 (0124) 6358322

2000

GATEKEEPER SERIES NO.SA93 1

 Executive Summary
The use of chemical pesticides and fertilisers in Indian agriculture has seen a
sharp increase in recent years, and in some areas has reached alarming levels
with grave implications for human health, the ecosystem and ground water. It is
therefore increasingly urgent that environmentally friendly methods of improv-
ing soil fertility and pests and disease control are used.

The potential of biopesticides and biofertilisers for promoting sustainable agri-

culture has been known for many years. A number of government agencies,
including the Ministry of Agriculture and the Department of Biotechnology, are
engaged in supporting research, production and application of these agents.
However, in spite of these efforts, their use in India is small. This paper investi-
gates the potential of and constraints in the use of biopesticides and biofertilis-
ers, taking the state of Haryana as a case study. It explores the factors responsi-
ble for the limited use of these agents, based on detailed discussions with a
large number of farmers, various agencies engaged in the promotion of biopes-
ticides and biofertilisers, State Agricultural Department officials, and shopkeep-
ers.

The study found that for the use of biopesticides, a key problem was that
departments promoting Integrated Pest Management (IPM) have very little
knowledge and experience of biopesticides, and most state agricultural universi-
ties, on whose recommendations pest control methods are promoted, do not
tend to recommend biopesticides. In the absence of active promotion by the
agriculture department, the demand for these products has not developed, and
most private shops and dealers do not stock and sell biopesticides. The paper
recommends that the agricultural departments and universities pay greater
attention to the promotion of biopesticides, that IPM training is improved, and
that there is a greater focus on cropping techniques and varieties which do not
require such a dependence on pesticides.

In the case of biofertilisers, their poor quality and performance is a major factor
in their limited uptake by farmers. This is primarily linked to inappropriate
strains and inefficient production technology. As a result it is recommended that
research and development to identify more suitable strains, to develop better
production technology and quality control methods is greatly increased, and
that in the meantime the various grants and subsidies on biofertilisers are divert-
ed to support these R&D programmes.

2 GATEKEEPER SERIES NO.SA93

A Study of Biopesticides and Biofertilisers in
Haryana, India
Ghayur Alam

Introduction
The use of chemical pesticides and fertilisers in Indian agriculture has seen a sharp
increase in recent years. In some areas, such as Haryana, Punjab and west Uttar
Pradesh, it has reached alarming levels. The heavy use of these chemicals has already
caused grave damage to health, ecosystems and ground water. It is therefore increasingly
urgent that environmentally friendly methods of improving soil fertility and pests and
disease control are used.

The potential of biopesticides and biofertilisers for promoting sustainable agriculture

has been known for many years. A number of government agencies, including the
Ministry of Agriculture and the Department of Biotechnology, are engaged in support-
ing research, production and application of these agents. However, in spite of these
efforts, their use in India is small. This paper investigates the potential of and constraints
in the use of biopesticides and biofertilisers, and explores the factors responsible for
the limited use of these agents. It is based on a study in the state of Haryana, a state
which represents the problem of excessive use of pesticides and fertilisers, common in
many parts of India. The paper also suggests policy measures for the promotion of
biopesticides and biofertilisers in the state. The study is based on detailed discussions
with a large number of farmers, various agencies engaged in the promotion of biopes-
ticides and biofertilisers, State Agricultural Department officials, and shopkeepers. The
study was carried out in 1999 as part of a research project on agricultural problems,
undertaken by the Agricultural Economics Research Centre, University of Delhi.

The Potential for Biopesticides

About 80,000 tons of pesticides are used in agriculture in India annually (Srinivasan,
1997), mostly in cotton and rice. While cotton is planted on about 5% of the total
cultivable area (on about 8 million hectares out of a total of 170 million), it accounts
for about 45% of pesticide application (Dhaliwal and Pathak, 1993). Rice accounts
for another 23%. Vegetables and fruit also account for a significant proportion (Table
1).

GATEKEEPER SERIES NO.SA93 3

 Table 1. Cropwise consumption of pesticide in India (%)

Cotton 44.5
Paddy 22.8
Jowar 8.9
Fruits and Vegetables 7.0
Wheat 6.4
Arhar 2.8
Other 7.6
Total 100.00
Source: Dudani and Sengupta, 1991

The intensive use of pesticides in agriculture is a cause of serious concern. The problem
is especially serious because of the development of resistance to pesticides in important
pests and the presence of pesticide residue in agricultural and dairy products.

Pesticide resistance in agriculture was first noticed in India in 1963 when a number of
serious pests were reported to have become resistant to DDT and HCH (two of the
most commonly used pesticides during the 1960s and 1970s). Since then the number
of pests with pesticide resistance has increased. The most serious problem of resistance
is witnessed in cotton, for which American bollworm is a serious pest. The bollworm
has developed resistance to almost all pesticides in a number of regions, and is partic-
ularly serious in parts of Punjab, Haryana, Andhra Pradesh, Karnataka and Maha-
rashtra. Other important pests of cotton, white fly and jassid, have also developed
pesticide resistance in some places.

Growing pesticide resistance has meant that a large proportion of agricultural produc-
tion is lost to pests. According to some estimates, these losses amount to between 20-
30% of total production (Mehrotra, 1989). The losses are particularly serious in cotton.
For example, cotton production in Punjab declined by about 50% during 1997 and
1998,1 causing a number of cotton farmers to commit suicide in the affected areas.2

Pesticide resistance has mainly been caused by excessive and indiscriminate use of pesti-
cides (Jayaraj, 1989). Pesticides of spurious quality, which are commonly sold in small
towns and villages, have also contributed to resistance in many areas. For example, in
Bidar (Karnataka) where the problem of pest resistance has become extremely serious,
more than 50 brands of pesticides were found to be sub-standard in 1998-99. In another
incident, the licenses of 115 pesticide producers were cancelled in Punjab for selling

1 “Another bad season for cotton farmers”, Hindustan Times, September 17, 1998.
2 For example, in Punjab 133 farmers were reported to have committed suicide in 1998 due to crop failure
caused by pest attack. “Another farmer in debt trap commits suicide”, Hindustan Times, October 4, 1998.

4 GATEKEEPER SERIES NO.SA93

sub-standard pesticide.3 Sub-standard pesticides contribute to resistance as the pests
are repeatedly exposed to a low concentration of pesticides. This contributes to the
build-up of resistance, without destroying the pests.

The other important problem caused by the excessive and inappropriate use of chem-
ical pesticides concerns the presence of pesticide residue in food. Many of the pesti-
cides currently being used have a tendency to survive in plants for a long time. They also
enter the food chain and are found in meat and dairy products. The problem of pesti-
cide residue is already a serious threat to health and environment in India. The inci-
dence of pesticide residue is much higher in India than in developed countries. For
example, according to one study, more than 80% of milk samples tested in India were
found to contain residues of DDT and HCH (Handa, undated). According to another
study, residue of DDT and benzene hexachloride, both suspected carcinogens, were
found in breast milk samples collected from mothers in Punjab. The amount of residue
was very high and babies were ingesting 21 times the amount of these chemicals consid-
ered acceptable through their mothers’ milk (Jumanah, 1994).

Compared to this, only 0.17% of samples tested in the US in 1990 were found to
contain residues over the acceptable limits.4 Similarly, in a study in the UK, only 1% of
the samples were found to contain residues above the prescribed limit.5

It is clear that the excessive use of chemical pesticides in agriculture is a serious cause
of concern. It is, therefore, important that alternative, environmentally friendly methods
of plant protection are adopted, such as integrated pest management (IPM) techniques,
including the use of biopesticides.

Biopesticides and Bio-control Agents

Biopesticides are derived from animals, plants and micro-organisms such as bacteria
and viruses. The advantages of biopesticides are:

• They are inherently less harmful than chemical pesticides;

• They are more target specific than chemical pesticides affecting only the target pests
and their close relatives. In contrast, chemical pesticides often destroy friendly insects,
birds and mammals.
• They are often effective in small quantities. Also, they decompose quickly and do not
leave problematic residues.

The most commonly used biopesticides include Bacillus thuringiensis (Bt), Baculoviruses
and neem. In addition to these, trichoderma, which is a fungicide, is also used. Bio-
control agents, such as Trichogramma, are parasites and predators of pests and their

3 “Another bad season for cotton farmers”, Hindustan Times, September 17, 1998.
4 “Current Pesticide Residue Levels in Food are Safe”, Pesticide Outlook (5)1, Feb 94. Cambridge.
5 “Latest UK Pesticide Residue Results Published”, Pesticide Outlook, Dec 1994, pp8. Cambridge.

GATEKEEPER SERIES NO.SA93 5

eggs. These biopesticides and bio-control agents are briefly described below:

• Bacillus thuringiensis (Bt). Bacillus thuringiensis is the most commonly used biopes-
ticide globally. It is primarily a pathogen of lepidopterous pests which are some of
the most damaging. These include American bollworm in cotton and stem borers in
rice. When ingested by pest larvae, Bt releases toxins which damage the mid gut of
the pest, eventually killing it. Bt based pesticides are being marketed by three compa-
nies in India. The total sale in 1999 was about 70 tons.6

• Baculoviruses. These are target specific viruses which can infect and destroy a number
of important plant pests. They are particularly effective against the lepidopterous
pests of cotton, rice and vegetables. Their large-scale production poses certain diffi-
culties, so their use has been limited to small areas. They are not available commer-
cially in India, but are being produced on a small scale by various IPM centres and
state agricultural departments.

• Neem. Derived from the neem tree (Azadirachta indica), this contains several chem-
icals, including ‘azadirachtin’, which affects the reproductive and digestive process
of a number of important pests. Recent research carried out in India and abroad has
led to the development of effective formulations of neem, which are being commer-
cially produced. As neem is non-toxic to birds and mammals and is non-carcinogenic,
its demand is likely to increase. However, the present demand is very small. Although
more than 100 firms are registered to produce neem-based pesticides in India, only a
handful are actually producing it. Furthermore, very little of the production is sold
locally, most being for export markets.

• Trichoderma. Trichoderma is a fungicide effective against soil born diseases such as

root rot. It is particularly relevant for dryland crops such as groundnut, black gram,
green gram and chickpea, which are susceptible to these diseases. Three companies
are marketing trichoderma in India.

• Trichogramma. Trichogramma are minute wasps which are exclusively egg-parasites.

They lay eggs in the eggs of various lepidopteran pests. After hatching, the
Trichogramma larvae feed on and destroy the host egg. Trichogramma is particularly
effective against lepidopteran pests like the sugarcane internode borer, pink bollworm
and sooted bollworms in cotton and stem borers in rice. They are also used against
vegetable and fruit pests. Trichogramma is the most popular bio-control agent in
India, mainly because it kills the pest in the egg stage, ensuring that the parasite is
destroyed before any damage is done to the crop. A number of countries produce
Trichogramma on a large scale. For example, in the former Soviet Union more than
10 biological factories were reported to produce about 50 billion Trichograma and
other parasites per season. Similarly, more than 50 commercial insectaries are reported

6 Dr MC Sharma, pers. comm. Director, Biotech International, New Delhi.

6 GATEKEEPER SERIES NO.SA93

 to be producing Trichogramma and other parasites in the USA and Canada. A number
of communes in China are also known to produce Trichogramma on a large scale.

Whilst India does not have technology to produce Trichogramma on a large scale, they
are being produced in small scale facilities for local use, mostly by sugar mills and coop-
eratives, state agricultural departments, IPM centres and agricultural universities.
Recently, some companies have started marketing Trichogramma through direct selling.
But the volume of sale is very small. Trichogramma eggs have to be used within a short
period (before the eggs hatch). This limits their production and marketing on a large
scale, and is also the reason why Trichogramma is not sold through dealers and shop-
keepers.

Promotion and effectiveness of Integrated Pest Management and

biopesticides
The Ministry of Agriculture and the Department of Biotechnology are responsible for
promoting biopesticides, the former via the Central IPM Centre (Faridabad), the
National Centre for IPM (NIPM) under the Indian Council For Agricultural Research
(ICAR) and the Directorate of Biological Control. As a part of the Department of
Biotechnology’s demonstration programme, biopesticides have been demonstrated on
about 55,000 hectares (Wahab, 1998). The Department has also supported a pilot plant
at the Tamil Nadu Agricultural University (Coimbatore) to develop and demonstrate
production and application technologies for baculoviruses, trichoderma and
Trichogramma.

Some Integrated Pest Management (IPM) demonstrations have shown success in

controlling pests without the use of chemical pesticides. For example, NIPM carried
out a demonstration in rice in a village in west UP on 100 acres in 1999. NIPM totally
substituted chemical pesticides with the bio-control agent Trichogramma, which is effec-
tive against both stem borer and leaf folder. According to NIPM, the control of pests
was complete and the yields were between 6-7 tons/hectare. Compared to this, the yield
in the area where chemical pesticides were applied was only 3.5 to 4 tons per hectare
(Damodran, 1999).

In another successful demonstration by NIPM on cotton in Maharashtra during 1998-

99, Baculoviruses, neem and Trichogramma were found to be more effective in control-
ling pests than chemical pesticides. The yield in these plots was about 1 ton/hectare,
compared to yields of only 300 to 500 kilograms/hectare in the fields where chemical
pesticides were used.7

7 “Integrated Pest Management Module for Dryland Regions - ICAR Trials in Cotton Fields Successful”.
Hindu Business Line, 4.1.1999.

GATEKEEPER SERIES NO.SA93 7

 The Potential for Biofertilisers
The growth in agricultural production during the last three decades has been accom-
panied by a sharp increase in the use of chemical fertilisers, causing serious concern
(Marothia, 1997). Foremost among these concerns is the effect of excessive fertiliser
(especially nitrogenous fertilisers) on the quality of soil and ground water.8

Biofertilisers are considered to be an important alternative source of plant nutrition.

They are biologically active products, including bacteria, algae or fungi, with the ability
to provide plants with nutrients. Most biofertilisers belong to one of two categories:
nitrogen fixing and phosphate solubilising. Nitrogen fixing biofertilisers fix atmos-
pheric nitrogen into forms which are readily useable by plants. These include rhizo-
bium, azatobacter, azospirillum, blue green algae (BGA) and azolla. While rhizobium
requires symbiotic association with the root nodules of legumes to fix nitrogen, others
can fix nitrogen independently. Phosphate solubilising micro-organisms (PSM) secrete
organic acids which enhance the uptake of phosphorus by plants by dissolving rock
phosphate and tricalcium phosphates. PSMs are particularly valuable as they are not
crop specific and can benefit all crops (Table 2).

Table 2. Major biofertilisers and target crops

Biofertiliser Target Crop

Rhizobium Leguminous crops
(Pulses, oilseeds, fodder)
Azatobacter Wheat, rice, vegetables
Azospirillum rice, sugarcane
Blue green algae (BGA) rice
Azolla rice
Phosphate solubilising microorganisms (PSMs) all

Production of biofertilisers in India

The idea of using micro-organisms to improve land productivity has been around in
India for at least 70 years, but it was only in the 1990s that large scale production of
various biofertilisers commenced. Presently, a number of agricultural universities, state
agricultural departments and commercial enterprises produce various biofertilisers.

The promotion of biofertilisers is mainly carried out by the National Biofertiliser Devel-
opment Centre (Ghaziabad), which was set up in 1987. The main objectives of the
National Centre are to:

8 Water containing excess of nitrates can affect the blood’s ability to transport oxygen, with serious health implica-
tions (WHO, 1963).

8 GATEKEEPER SERIES NO.SA93

• produce and market biofertilisers of required quality;
• isolate and maintain biofertiliser strains suitable to various agro-climatic regions;
• train agricultural extension workers;
• promote biofertilisers through field demonstrations;
• prepare quality parameters;
• test samples of biofertilisers produced by others;
• provide technical and financial assistance to units producing biofertilisers.

The National Centre has the capacity to produce 375 tons of biofertilisers per year. In
addition to this, 58 commercial production units have been set up with government
support. India’s total production in 1998-99 was reported to be 16,000 tons.9 Rhizo-
bium accounts for the largest proportion (40%) of the total production in India. This
is followed by azatobacter. With the increase in the price of phosphate fertilisers, the
potential for the use of PSM has also increased.

Effectiveness of biofertilisers
A considerable amount of research has been done to establish the effectiveness of biofer-
tilisers on various crops, in different agro-climatic regions. Most agricultural universi-
ties, the ICAR and the National Biofertiliser Development Centre have carried out a
number of field trials to document the effectiveness of these micro-organisms. These
programmes show that the use of biofertilisers can have a significant effect on the yield
of most crops. However, their effectiveness is found to vary greatly, depending largely
on soil condition, temperature and farming practices. As an example, Table 3 shows the
effect of azatobacter on yield.

Table 3. Effect of azatobacter on crop yield

Crop Increase in yield over Crop Increase in yield over

yields obtained with yields obtained with
chemical fertilisers (%) chemical fertilisers (%)
Food grains Other
Wheat 8-10 Potato 13
Rice 5 Carrot 16
Maize 15-20 Cauliflower 40
Sorghum 15-20 Tomato 2-24
Cotton 7-27
Sugarcane 9-24
Source: Das, 1991

9 Dr. T. Singh, pers. comm., Director, National Biofertiliser Development Centre.

GATEKEEPER SERIES NO.SA93 9

 The Case Study:
Karnal and Bhiwani Districts, Haryana
A field study was initiated in two districts in Haryana to try to understand why biofer-
tilisers and biopesticides were not being adopted on a wide scale. The study focused
on two districts: Karnal and Bhiwani. While Karnal represents intensive agriculture
with a high degree of irrigation, Bhiwani represents dryland farming, with low level
irrigation. The potential for biofertilisers and pesticides is described below, followed
by the results of our interviews with farmers to establish why biofertilisers and biopes-
ticdes have not been taken up more widely.

The Potential for Biopesticides and Biofertilisers in Haryana

The problems caused by excessive pesticide use are particularly serious in rice and
cotton in Haryana, both important crops in our study districts. The American boll-
worm, jassid and white fly in cotton; and stem borer in rice; have developed resistance
to chemicals in many parts of Haryana, including parts of the districts studied. In this
situation there is great potential for biopesticides.

The potential of biofertilisers is evidenced by the fact that about 90% of Haryana’s soil
is deficient in nitrogen, indicating severe nutrient deficiency (Dahiya et al., 1993). The
dryland districts such as Bhiwani, Mohindergarh, Sirsa and Faridabad are particularly
low in nitrogen, and soils are short of organic matter due to poor vegetative cover, high
temperatures and the light texture of soil. Biofertilisers could play a role in providing
the much needed nutrients and improving soil conditions in these dryland areas, which
account for about 28% of the state’s total cultivable area.

Biofertilisers can also reduce the intensity of chemical fertiliser consumption, especially
in irrigated areas. With the increase in cropping intensity (see below), the use of chem-
ical fertilisers has increased significantly in these areas. The use of organic manure, on
the other hand, has not increased. As a result, many parts of Haryana face deteriorat-
ing soil conditions and increasing ground water contamination.

The suitability of biofertilisers for various crops grown in Haryana has been shown
through demonstrations conducted by the Hissar Centre of the National Biofertiliser
Development Centre (NBDC). Azatobacter and azospirillum, which can be used for a
wide range of crops, are estimated to have particularly large potential. For example,
NBDC experiments showed an increase in yield between 3 and 25% with the applica-
tion of azotobacter in cotton. Similarly, in the case of wheat, use of azotobacter resulted
in yield increases between 2 and 20%.10

10 National project on use and development of biofertilisers. Biofertiliser News, 1(2), December 1993.

10 GATEKEEPER SERIES NO.SA93

The potential for the use of rhizobium is largely confined to the dryland areas of
Haryana, where pulses are commonly grown, and where the soils show poor to moder-
ate nodulation.11

Karnal District
In Karnal the main nutrient and pest problems relate to the district’s high cropping
intensity. Wheat-rice rotation has been common for some time now, and cropping inten-
sity has recently increased further with the introduction of short duration rice varieties
such as Govinda (90 days). Planting these varieties allows farmers to take two rice crops
between April and October. The first paddy crop is planted in April-May, as soon as
wheat has been harvested. This rice crop is ready for harvest by mid July. This is
followed immediately by the transplanting of fresh paddy seedlings. The second rice
crop is harvested by the end of October, to be followed by wheat, which is sown by
early November.

This very high intensity of cropping has worsened both the soil quality and pest problem
in parts of Karnal. It is extremely damaging to the soil condition, as large amounts of
nutrients are used continuously without replenishment. As a new crop is planted as
soon as the older crop is harvested (sometimes both activities are done simultaneously),
there is no time for proper land preparation and for using organic manure. As the use
of chemical fertilisers has increased to provide the required nutrients, soil and water
conditions have deteriorated.

The problem of pests and pesticide use in Karnal is largely confined to rice. Growing
two rice crops without a break is one factor. The continuation of rice plants in the same
field, and the high degree of moisture, enable pests to multiply without a break, leading
to particularly intense pest attacks in the second crop. Another reason is the popular-
ity of certain basmati varieties which are highly susceptible to pests and diseases. This
is particularly serious in one of the most popular of these varieties, called duplicate
basmati. This variety was introduced about five years ago from West UP, and became
very popular because of its high yields,12 and totally replaced the desi (traditional)
basmati variety in many areas. After performing well for three years, the variety began
to be affected by pests (stem borer, leaf folder and white back plant hopper) and diseases
(sheath blight, blast and bacterial leaf blight) two years ago. The attack was particu-
larly severe last year, forcing many farmers to stop planting basmati in general, and
duplicate basmati in particular. In one of the villages we studied, Kuuchhpura, farmers
made up to 15 pesticide applications last year but could not save the crop. This year,
very few farmers have planted basmati in the village.

11 The assimilation of atmospheric nitrogen by the roots of pulse (and other leguminous) crops is carried out
by the nodules formed in the roots of these plants. The low degree of nodulation suggests that their ability to
assimilate atmospheric nitrogen is low and that they could benefit from the use of rhizobium.
12 Compared to the average yield of 10 quintal/acre from desi basmati, the yields of duplicate basmati were
about 20 quintal/acre.

GATEKEEPER SERIES NO.SA93 11

The problem of pesticide resistance in stem borer is now common in Karnal, largely
caused by the indiscriminate and excessive use of pesticides. Another important reason
relates to the fact that farmers in Karnal do not spray their fields themselves. This is
done by professional workers who are sent by the shops selling pesticides. As these
workers are paid by the area covered, they do a rushed job. The spraying is not uniform:
while some parts of the fields get very little, others get excessive pesticides. Secondly,
in order to save time they use about one third of the water prescribed to make the pesti-
cide solution. The non-uniform spray of highly concentrated pesticide solution is
reported to be a major reason for the development of resistance in rice pests in many
parts of the district.13

Bhiwani District
In Bhiwani, the focus of the study is on cotton and gram, which, along with wheat and
mustard are the main crops in the area. The use of chemical fertilisers is comparatively
low in Bhiwani, being used only on wheat and cotton, and not at all on gram and
mustard. But, as in Karnal, use of organic manure is not common and the condition of
the soil is poor. Three biofertilisers have potential for Bhiwani: rhizobium for gram and
azatobacter and PSM for wheat. The potential of rhizobium is reported to be particu-
larly high in Bhiwani because the level of nodulation is very low.

The District Agriculture Department is responsible for the sale of rhizobium in Bhiwani.
Being a major gram growing district, it was allocated the largest amount of rhizobium
last year and its sale to farmers is subsidised at the rate of 50%.

The main pest problem in Bhiwani concerns cotton, which is attacked by American,
pink and spotted bollworm, white fly and jassid. These pests have become resistant to
most pesticides. According to a study of pesticide use in Bhiwani in 1998, about 70%
of farmers reported that they were unable to control pests with pesticides (Saini and
Jaglan, 1998). Some of these farmers had used up to 11 pesticide applications.

The pest problem in Bhiwani is closely linked with the spread of irrigation facilities.
Large parts of Bhiwani district are irrigated by the Indira Gandhi Canal, which suffers
from large scale seepage. This has led to a rise in the groundwater level from 50-60 feet
below the surface in the past to 5-10 feet at present in many areas. As a result, water
logging and high humidity are serious problems in many areas of the district. Apart
from causing direct damage, this has also created favourable conditions for the growth
of pests. Consequently, the problem of pests has become extremely serious in the district
during the last five years.

The pest problem caused the area under cotton to decline in the second half of the
1990s from 57, 000 hectares in 1996-97 to 51,000 hectares in 1999-2000.14 As in rice

13 Information from the District Agricultural Officers.

14 Information provided by the State Agricultural Department, Bhiwani.

12 GATEKEEPER SERIES NO.SA93

in Karnal, the pest problem in cotton in Bhiwani is closely related to the susceptibility
of the varieties being planted. The cultivation of cotton in this area is comparatively
new: it was introduced in the 1970s. Most of the varieties introduced since then are
long staple, American types. Although they fetch high prices, they are highly suscepti-
ble to various cotton pests. The problem of pests and the excessive use of pesticides in
Bhiwani is mostly confined to the American varieties. Desi varieties, which are compar-
atively resistant to pests and diseases, require less use of pesticides.

We observed a shift back in favour of desi cotton in one of the divisions of Bhiwani
following severe losses to pests to the American varieties. Whilst many more farmers
would prefer to shift to desi cotton varieties, as they provide stable yields, their whole-
sale prices are too low. In addition it appears that the state agriculture department
prefers to promote long staple American varieties, as they have large domestic and
export markets.

Farmers’ Perspectives
We selected 11 villages from the two districts, six villages from Karnal and five from
Bhiwani, in which to interview farmers to establish their awareness and use of biofer-
tilisers and biopesticides. A total of 74 farmers from these villages were interviewed, and
were mainly selected because they had attended one of the demonstration programmes
carried out by government agencies to promote IPM, biopesticides and biofertilisers
(58 of the farmers, or about 80% of the sample). However, some farmers who had not
been to the programmes were also interviewed (16 farmers). All the farmers were men;
only men had participated in the demonstration programmes, and we were told that
men take the decisions about the types of fertilisers and pesticides to be used.

The farmers included in the study represent the range of small, medium and large
farmers. While 29% of the farmers have less than 5 acres of land, 46% have between
5 and 20 acres. 25% of the farmers have landholdings larger than 20 acres. In terms
of education, about one third of the farmers have studied up to class 10 or more. A
semi-structured questionnaire was used for interviews, which were conducted in
farmers’ homes.

Findings: Biopesticides
Although biopesticides and bio-control agents are important components of IPM, the
IPM programmes being conducted by various agencies put very little emphasis on these
agents. None of the farmers we interviewed had ever used biopesticides, and few were
even aware they existed. We found that none of the farmers had even used neem, widely
believed to be commonly used by Indian farmers.

In fact, we found that IPM itself was not being practised by most farmers. While some
of the farmers were aware of IPM practices, such as the need for monitoring and

GATEKEEPER SERIES NO.SA93 13

augmenting natural enemies of pests, very few farmers have adopted these practices.
None of the farmers practised IPM fully, but about 15% practised partial IPM. In most
cases this meant delaying the first spray of pesticides until damage by pests becomes
evident. The situation is particularly bad in Bhiwani, where 85% of farmers reported
not using IPM at all and only 9% reported using some aspects of IPM. Compared to
this, 20% of farmers reported using limited IPM in Karnal. This is despite the fact that
most of the farmers included in the study participated in the IPM demonstration
programmes conducted by the Central IPM.

Why is IPM not practised?

We found four important reasons for the low acceptance of IPM:

1. Lack of awareness. This reason was found to be particularly important in Bhiwani,

where 77% of farmers were unaware of the concepts of IPM. Compared to this, fewer
than 3% of farmers in Karnal were unaware of IPM. Clearly, there is a big district-
wise difference in the success of IPM demonstration programmes and state agricultural
extension workers in familiarising farmers with IPM. In Karnal these agencies have
been successful in at least making farmers aware of the need to practise IPM.

2. Lack of skills. Almost all the farmers, including all of those who were aware of IPM,
reported that they lacked the skills necessary to practise IPM. Their ability to practise
IPM was, for example, severely constrained by the fact that most of them could not differ-
entiate between harmful and beneficial insects. In fact, many farmers thought that all
insects were a potential threat to their crop and had to be destroyed. They were also not
able to work out economic thresholds15 to determine the timings of pesticide application.

3. Lack of faith in IPM. This factor was found to be very important amongst 60% of
farmers in Karnal. Although almost all of them were aware of IPM, they felt that they
did not have sufficient faith in it to reduce the use of chemical pesticides. Many of
them felt that the support from the agricultural department was not adequate for them
to try IPM practices, which were considered risky. As the CIPM personnel do not keep
in touch with the farmers after IPM demonstrations, they felt that they would not get
adequate advice and support if things went wrong. Similarly, the local extension
workers (ADOs) are not sufficiently trained in IPM to instil confidence in the farmers.

The issue of skills and confidence is obviously linked to the intensity of training
provided by the IPM agencies to farmers and extension workers. We found that the
training is very basic and superficial, being conducted for three to four days in a
village, during which 30 villagers and five extension workers are trained. The farmers
felt that the training was not intensive and did not impart sufficient skills for them
to feel competent and confident in following IPM practices.

15 The economic threshold is the level of pest population at which the damage to crop justifies the use of pest
control methods.

14 GATEKEEPER SERIES NO.SA93

4. IPM practices are difficult and cumbersome. About 70% of the farmers in both
districts who were aware of IPM, felt that the IPM practices were too cumbersome
and time consuming to be used regularly. Both the monitoring of pest populations
and the calculation of economic thresholds were considered by farmers to be imprac-
tical. In Karnal, where cropping intensity is high, the farmers felt that they did not
have time to keep a close watch on their fields to monitor pests and calculate
economic thresholds.

Whose advice is taken?

An important reason for the failure of the IPM programme, and the lack of biopesti-
cide use, is related to the fact that most farmers depend on shopkeepers for advice on
pesticide application. More than 80% of the farmers reported that shopkeepers, dealers
and representatives of pesticide manufacturers were their most important sources of
information about pest control methods (Table 4).

Table 4. Importance of source of advice for pesticide application

Other Extension Shops/manufacturers/

Farmers workers and dealers
agricultural university
Karnal 12 (30) 18 (45) 33 (83)
Bhiwani 10 (30) 9 (26) 27 (80)
Total 22 (30) 27 (36) 60 (81)
Number of responses=74. Note that some farmers listed more than one source of advice as being
equally important
Note: Figures in parenthesis indicate percentages.

Compared to the manufacturers and sellers of pesticides, agricultural extension workers

play a small role, especially in Bhiwani, where only 26% of farmers reported them to
be important.

The role of shopkeepers and dealers is particularly important because many farmers
(58%) purchase their pesticides on credit. This gives the shopkeepers strong control
over the amount and choice of pesticides used. It also makes it easier for the shop-
keepers to sell spurious pesticides.

Finally, we found that the knowledge of the State Agricultural Department about
biopesticides was extremely limited. This was particularly true of the village level
workers (ADOs), but also the case even in the Central IPM office in Faridabad. Consid-
ering the importance given to the use of biopesticides by national government agen-
cies, the neglect of biopesticides at the state and district level is difficult to understand.
The main emphasis is on the monitoring of pest populations and the use of economic
thresholds, which farmers find too difficult and time consuming. Further, the Haryana

GATEKEEPER SERIES NO.SA93 15

Agricultural University recommends the use of only two biopesticides (Bt and neem) on
only one vegetable crop. On all other crops, including cotton and rice, the recommen-
dations include only chemical pesticides.

Findings: Biofertilisers
Few (19%) farmers have ever used biofertilisers in the two districts. Their number was
especially small in Karnal, where only three out of 40 farmers had used them. In
Bhiwani about 75% of farmers were not even aware of biofertilisers; the proportion of
such farmers was even higher in Karnal (85%).

The most important reason for this lack of awareness is the fact that agriculture exten-
sion workers do not promote biofertilisers. On the whole, only 15% of farmers had
been told about biofertilisers by the extension workers. The emphasis on biofertilisers
was particularly low in Karnal, where only one farmer out of 40 had heard about them
from extension workers.

The District Agricultural Departments do not have a positive attitude towards biofer-
tilisers. They feel that their quality is poor, and their performance totally unreliable.
Therefore, they are not prepared to risk their reputation and good will with the farmers
by recommending them. The extension services run by the Haryana Agriculture Univer-
sity (krishi vigyan kendra) feel that in areas of wheat/paddy rotation, such as Karnal,
the potential of biofertilisers is low. The farmers in these areas can get the same yield
by using the recommended dose of chemical fertilisers. As these fertilisers are easy to
use, the farmers prefer them. Biofertilisers have to be stored and applied in conditions
which are suitable for the multipication of micro-organisms. This requires special facil-
ities and care, which farmers are often unable to provide. Chemical fertilisers, on the
other hand, can be stored and applied without special care. In fact, the KVK in Karnal
does not recommend the use of biofertilisers at all.

Nevertheless, the extension workers in Bhiwani are required to sell a fixed number of
rhizobium packets. This explains the larger number of farmers who are aware of biofer-
tilisers, and have used them in this district. But we found that a large proportion of the
rhizobium allotted to the district is not sold to the farmers, and is allowed to go to
waste. The ADOs say that the quality and performance is so poor that the farmers are
not interested in buying it. The quota is shown as sold in official records, and the
payment is made by the ADOs from their salary.16 The fact that the rhizobium meant
for sale in various pulse growing districts is not being used by farmers is widely known
to both the State Agricultural Department and the National Biofertiliser Development

16 For example, one of the ADOs in Bhiwani was given a quota of 2000 packets of rhizobium this year. He
could manage to sell only about 200 packets, mostly to his contact farmers. But these have not actually been
used by the farmers. He feels that the contact farmers accept biofertilisers because they want to stay in the good
books of the ADOs, from whom they receive subsidised or free goods, such as seed kits. He will have to pay
to the Agricultural Department Rs. 8,000/- (@Rs. 4.00 a packet).

16 GATEKEEPER SERIES NO.SA93

Centre. However, every year the practice of fixing quotas and reporting of sale is carried
out, as if rhizobium is actually being used.

Out of 19 farmers who were aware of biofertilisers, 14 had used them, showing that
farmers are prepared to try biofertilisers. However, most of these farmers stopped using
them after one crop - only three were still using them. Two reasons were reported to
be important for the discontinuation of use: lack of availability and poor performance.

Availability
Many farmers who stopped using biofertilisers reported that this was partly because the
supply was extremely unreliable. This was largely because biofertilisers were not being
sold by most shops. While none of the shops stock biofertilisers in Karnal, two shops
in Bhiwani do. The shopkeepers, in turn, say that they do not stock biofertilisers
because sales are poor. One of the Bhiwani shops, for example, has been stocking biofer-
tilisers from National Fertilisers Ltd. for the last four years but has sold only 30 packets.

Quality
It is clear that the poor quality and performance of biofertilisers present serious prob-
lems. Most studies suggest that the biofertilisers being sold in the market are contam-
inated and have a low count of micro-organisms. It is therefore not surprising that their
performance is poor and uneven. For example, in a survey of rhizobium carried out by
ICRISAT, 90% of samples from India were found to have a rhizobia count lower than
that required for effective performance (Singleton et al., 1996). Incidentally, this
problem exists in most developing countries. For example, in a survey of 12 develop-
ing countries, only 19% of the samples met the standards prevalent in developed coun-
tries (Singleton et al., 1996).

The poor performance of biofertilisers in India is primarily linked to inappropriate

strains and inefficient production technology. Essentially, the production of bacterial
biofertilisers requires the selection of strains appropriate for a particular crop in a given
agro-climate. These strains are mass multiplied by adding bacterial culture to a suit-
able sterilised broth, either using the shake flask method (for small scale production)
or the fermenter method (for large scale production). When an adequate growth of
bacteria is achieved, the solution is mixed with a carrier such as lignite or charcoal and
is packaged.

As agro-climatic conditions and soil characteristics vary widely, a large range of strains
of each biofertiliser needs to be isolated for each area. The problem of identifying suit-
able strains is particularly serious in north India, as many of the strains do not survive
the very hot temperatures prevalent in these areas. Until strains which can tolerate wide
variations in temperature can be identified, the performance of biofertilisers will remain
uneven. The Haryana Agriculture University is reported to be working in this direction
and has developed improved strains for pearl millet, wheat, mustard, potatoes, and
flowering plants. These are, however, yet to be used in large scale production.

GATEKEEPER SERIES NO.SA93 17

Furthermore, the production of biofertilisers is prone to contamination, which reduces
the effectiveness of micro-organisms. It is, therefore, vitally important that throughout
the process extreme care is taken to maintain sterile conditions. It is also important
that precautions are taken to avoid contamination during the packaging, storage and
application of biofertilisers.

The production technology employed in India is inefficient and is responsible for most
of the contamination common in Indian biofertilisers. Generally, production is under-
taken by the flask method, which is unsuitable for large scale production. Although
some firms use fermenters, they lack the sophisticated controls and monitoring facili-
ties necessary to regulate factors such as pH, temperature and aeration. As a result the
quality of the bacterial broth is often poor and uneven.

Another problem relates to the fact that Indian producers do not sterilise the carriers
used for mixing the bacterial solution. For example, both the producers in Haryana, the
Hissar Biofertiliser Centre and the HAU, use non-sterilised charcoal, as they do not
have facilities to sterilise large amounts of charcoal in a short time. This is an impor-
tant cause of the poor quality and short shelf life of these biofertilisers (Singh et al.,
1999). Although India has ISI (Indian Standards Institution) standards for some of the
biopesticides (rhizobium and azatobacter), they are not enforced. This is reportedly
because the ISI lacks facilities to test biofertilisers.

Conclusions
Biopesticides
The problem of pests, the development of pesticide resistance and the excessive use of
pesticides need to be seen in the totality of the agricultural system. Our study shows that
in Karnal and Bhiwani the problem is linked to the increase in cropping intensity (three
crops in Karnal), the expansion of irrigation facilities (Bhiwani), the release and adop-
tion of susceptible varieties (govinda and basmati rice in Karnal, and American cotton
varieties in Bhiwani), purchase of pesticides on credit (in both the districts) and inap-
propriate agricultural practices (the use of contract labour for pesticide application,
using power spraying machines in Karnal). In the circumstances, mere reliance on pest
control, without correcting the basic problem in the system, will not produce sustain-
able results.

The efforts of various government agencies to popularise integrated pest management

(IPM), and the use of biopesticides have had little impact. IPM departments have very
little knowledge and experience of biopesticides, and most state agricultural universi-
ties, on whose recommendations pest control methods are promoted, do not include the
use of biopesticides in their recommendations. In the absence of active promotion by
the agriculture department, the demand for these products has not developed. It is for
this reason that most private shops and dealers do not stock and sell biopesticides.

18
18 GATEKEEPER SERIES NO.SA91
NO.SA93
The following policy measures need to be taken urgently in order to reduce excessive
use of chemical pesticides.

• Focus on sustainable agriculture by promoting: a) disease and pest resistant, and espe-
cially traditional, varieties; b) judicious inter-cropping and c) reduced crop intensity.
• Improvement in the intensity of training for IPM. The focus should be on the quality
of training and not on the number of farmers trained. The training should be followed
by regular contact with the trained farmers for providing continous support.
• Promotion of the use of biopesticides by the state agricultural departments and IPM
workers. The state agricultural universities, which have decisive influence over what
pest control methods are promoted by governmental agencies, should pay greater
attention to biopesticides.

Biofertilisers
Despite the Indian government’s efforts to promote the production and use of biofer-
tilisers, our study found that biofertilisers have found little acceptance among farmers
in Haryana. The problems of unavailability of biofertilisers and their poor quality are
linked. On the one hand, we find that both the State Agriculture Department and shop-
keepers are unwilling to stock and sell biofertilisers as they feel that their quality is
unreliable. On the other hand, the low demand for biofertilisers has prevented large
investment in advanced production and storage facilities, which are required for
improving the quality. It is, therefore, clear that the two problems have to be seen in
their totality, and a new policy is needed, some elements of which are as follows:

• The present policy of providing grants and low interest loans to biofertiliser produc-
ers should be abolished; this has resulted in the setting up of a large number of inef-
ficient plants, which cannot produce good quality biofertilisers.

• The policy of marketing biofertilisers at very low prices should also be stopped. These
prices are too low to attract adequate investment in modern manufacturing facilities.
Take the example of rhizobium. A 200 gram packet of biofertiliser, which is supposed
to replace about 30 kilograms of urea (a commonly used chemical fertiliser), is
purchased by the Agricultural Department for Rs. 8/ packet. This price is not enough
to justify investment in facilities such as charcoal sterilisation plants and cold chain
for storage and transportation. Rhizobium is sold to farmers at a subsidised rate of
Rs. 4/-. Our discussions with farmers suggest they feel that nothing so cheap can
provide much nutrition to the plants, and do not value it. The price of biofertilisers
should have some relationship with the price of the chemical fertiliser it replaces.
Only then will the producers, the shopkeepers and the farmers begin to take biofer-
tilisers seriously.

• The storage and application of biofertilisers require special facilities and skills, which
most producers, shopkeepers and farmers do not possess. It is important that greater

GATEKEEPER SERIES NO.SA93 19

 R&D efforts are focused on developing biofertilizers which are easier to store and
apply.

• Research and development to identify more suitable strains, and to develop better
production technology and quality control methods has to be increased greatly. The
money saved through the abolishment of various grants and subsidies should be
invested in these R&D programmes.

We recommend that the promotion and production of biofertilisers should be suspended

until these steps are taken. The present government policy of promoting biofertilisers
without ensuring good quality and performance has actually harmed their cause, creat-
ing a widespread feeling among farmers and extension workers that biofertilisers do
not work. It will be better if the production and promotion of biofertilisers is suspended
until biofertilisers of improved quality can be made available in adequate quantity. Until
then all efforts should be focused on developing technology, setting up modern produc-
tion facilities and developing infrastructure that will produce and deliver biofertilisers
of the required quality.

Acknowledgements
The study was funded by the Agricultural Economics Research Center, University of
Delhi. The author is grateful to Professor Prem Vashishtha, Director, Agricultural
Economics Research Center, University of Delhi for his support to the study. The views
expressed in the paper, however, are solely the responsibility of the author.

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Methods. 1993. G. Gill.
39. Homegarden Systems: Agricultural Char-
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40. Opportunities for Expanding Water Harv-
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41 Out of Print
42. Community First: Landcare in Australia.
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23
71. Propelling Change from the Bottom-Up: 85. Sharing the Last Drop: Water Scarcity,
Institutional Reform in Zimbabwe. 1997. J. Irrigation and Gendered Poverty Eradication.
Hagmann, E. Chuma, M. Connolly and K. 1999. Barbara van Koppen.
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24 GATEKEEPER SERIES NO.SA93


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ISSN 1357-9258
Nutr Cycl Agroecosyst (2009) 84:167–178
DOI 10.1007/s10705-008-9235-6
Dissolution of phosphorus from animal bone char in 12 soils
G. P. Warren Æ J. S. Robinson Æ E. Someus
Received: 1 May 2008 / Accepted: 28 November 2008 / Published online: 14 December 2008
Ó Springer Science+Business Media B.V. 2008
Abstract Heat-treated animal bone char (ABC) has
not previously been evaluated for its potential as a
phosphorus (P) fertilizer. ABC, Gafsa phosphate rock
(GPR) and triple superphosphate fertilizer (TSP)
were incubated in 12 soils. Dissolved-P was assessed
by extraction with NaOH and bioavailability with the
Olsen extractant. The rate of P dissolution from ABC
was described almost equally well by the Elovich and
Power equations. After 145 days, the fraction of P
dissolved ranged from 0 to 73% and to 56% for ABC
and GPR, respectively. The most important soil
properties determining P dissolution from ABC were
pH and P sorption. P dissolution was not significant at
soil pH [6.1 (ABC) and [5 (GPR) and the lower the
pH, the greater the Dissolved-P. Dissolved-P also
correlated positively and significantly with inorganic
P sorption, measured by the Freundlich isotherm and
the P sorption index of Bache and Williams (1971).
Soil pH and P sorption index could be combined in
multiple regression equations that use readily
G. P. Warren (&)
J. S. Robinson
Department of Soil Science, The University of Reading,
P.O. Box 233, Reading RG6 6DW, UK
e-mail: g.p.warren@reading.ac.uk
E. Someus
Terra Humana Ltd, Széchenyi u. 59,
1222 Budapest, Hungary
measured soil properties to predict the potential for
ABC dissolution in a soil. Dissolution of P from GPR
correlated with soil pH and exchangeable acidity. In
comparison with GPR, ABC was a better source of
available P, assessed by Olsen-P. In most soils, ABC
increased Olsen-P immediately after application,
including soils of relatively high pH in which GPR
was ineffective. ABC is a P fertilizer of solubility
intermediate between GPR and TSP.
Keywords Available P
Biochar

Fertilizer
Phosphorus
Phosphate rock
Introduction
It is probable that bones have always been regarded
as a valuable manure, and by about the end of the
eighteenth century, their manurial value was attrib-
uted to phosphates (Smith 1959). Bone products
remain advocated and widely used in domestic
gardening (Royal Horticultural Society 2002). How-
ever, by the 1950s, for most commercial farming,
bone and P fertilizers derived from it were sup-
planted by fertilizers made from mineral deposits
(Smith 1959). This lack of interest in bone as a
fertilizer for commercial purposes may explain the
observation that reliable data on the plant availabil-
ity of P from recycled products of P-containing
wastes such as sewage sludge, meat and bone are
123
168
rare (Romer 2006). Nevertheless, trials in arable
fields can show a slow release of P from bone,
comparable to that of phosphate rock (PR) directly
applied to soil (Bekele and Hofner 1993) or, in
certain soils, an effectiveness as good as superphos-
phate fertilizer (Ramos 1982). Reasons for looking
at alternative and renewable sources of P include
the future depletion of economically recoverable PR
reserves and the need to restrict introduction of
toxic metals to agricultural soil via fertilizer, for
which bone is a clean P source. For example,
reviews show that cadmium concentrations range up
to only 3.03 mg kg-1 in cattle and pig bone (Doyle
1979) but up to 556 mg kg-1 in PR (Van Kauwen-
bergh 1997). However, concerns have arisen about
the transmission of diseases through animal by-
products and in the European Union, regulations
control their disposal (EU 2002). Controlled thermal
treatment exceeding about 400°C can be used to
make a form of biochar that is free of infective
agents, rich in P, and that we call animal bone char
(ABC). ABC appears potentially an effective P
fertilizer, but it has not been tested for this purpose.
To assess its value and limitations as a P fertilizer,
information is required about its rate of dissolution
in soil and soil properties that may influence its
dissolution.
A substantial body of information exists on the
dissolution rate of P from PR directly applied to soil.
The requirements for P dissolution are a source of
acidity, and sinks for P and Ca (Rajan et al. 1996),
which must be provided by the soil/plant system.
Sedimentary carbonate fluorapatites are the PRs that
are most widely exploited for agriculture (Khasawneh
and Doll 1978). Chemical and X-ray diffraction
evidence show that the inorganic phases of bone
should also be classed as carbonate apatites (LeGeros
1994), so it appears that P dissolution from bone,
ABC and phosphate rock should be controlled by the
same soil factors. However, this hypothesis has not
been tested. The first aim of the work was therefore to
measure the dissolution of P from ABC, in compar-
ison with well-known soluble and slowly-soluble P
fertilizers, relate P dissolution from ABC to a wide
range of soil characteristics, and test the hypothesis
that P dissolution is controlled by the same soil
properties for ABC as for PR. The second aim was to
assess the ability of ABC to improve plant-available
P in soil.
123
Nutr Cycl Agroecosyst (2009) 84:167–178
Materials and methods
Soils
Soils were obtained from 12 locations, chosen to
provide a range of types and to include some with
severe P deficiency. Sampling was from the 0 to
20 cm horizon using multiple auger holes or pits.
Stones were removed with a coarse sieve (approx
2 cm aperture), the soil air-dried for 2 or 3 days at
room temperature or in a well-ventilated oven at
30°C and the soil ground to pass a sieve of 2 mm
mesh. Table 1 gives their main properties.
P sources
The ABC (batch SM7) was supplied by Terra
Humana Ltd. It was derived from cattle bone, and
prepared by heating at 400°C for 45 min. Gafsa
Phosphate Rock (GPR), from Tunisia, was a carbon-
ate fluorapatite containing some accessory calcitic
materials (Hammond et al. 1986). A fully soluble
fertilizer, Triple Superphosphate (TSP, containing
20% P), was used to provide a benchmark for
agronomic effectiveness, against which to compare
the slowly soluble sources, and to assess the potential
of soils for immobilization of free phosphate. To
minimise the potential effect of particle size, all P
sources were crushed gently in a pestle and mortar to
pass a 2 mm aperture sieve.
Analytical methods
For the ABC and PR, P extracted by neutral
ammonium citrate was measured by the method of
Mackay et al. (1990), P extracted by water was at a
solid:solution ratio of 1:50 for 1 h at 20°C, and
surface area was measured by gas adsorption. Mea-
surements of Olsen-P (0.5 M NaHCO3 adjusted to pH
8.5), pH in water and 0.01 M CaCl2, exchangeable
acidity, organic C and clay were made as described
by Rowell (1994). Total C was measured by
combustion (Leco model SC444 analyser) and total
P by dissolution in H2SO4/H2O2/Li2SO4/Se solution
at 360°C and colorimetric analysis (Anderson and
Ingram 1993). CEC and exchangeable cations were
measured by the unbuffered method of Sumner and
Miller (1996). pH in KCl and pH buffer capacity
Nutr Cycl Agroecosyst (2009) 84:167–178 169

Grass Fallow

Grass Fallow
were measured by the method of Robinson et al.

Veg/Grass
Vegetable

Vegetable
Vegetable

Vegetable
(1992). The P sorption index of Bache and Williams
Land use

Arable

Arable

Arable
Forest

Forest
(1971) was measured, being the quotient X/logC,
where X equals P sorbed (mg P 100 g-1 soil) and C
the P concentration in solution (lmol P l-1). Two
Ca (cmol kg-1)

indices were measured, one with P addition at the

Exchangeable

usual P addition rate of 1.5 g P kg-1 (Pindex-1500)

and also at a low rate of 300 mg P kg-1 (Pindex-300),
11.51

21.96
26.45

13.88
2.84

3.95
0.29
1.14
1.24
3.02
6.83

1.05
comparable to the actual rate of added P used in the
incubation experiment. To measure the P sorption
(cmol kg-1)

isotherm, samples of soil were equilibrated with end-

over-end shaking for 16 h at 20°C in 0.03 M KCl
3.72
4.25

7.91
1.23
1.92
3.13
5.37

5.41
35.80
37.19

12.53
21.89
CEC

(soil:solution = 1:20), with P additions to result in

final concentrations in the range 0 to 20 mg P l-1. In
many soils the amount of readily extractable P was
acidity (cmol kg-1)

high and so there was substantial desorption of soil P

Exchangeable

at low initial solution P concentrations, making it

difficult to fit equations to describe the P sorption
0.044
0.075
0.063
-0.022
0.044
0.615
0.144
0.298
0.282
0.081
-0.010
5.152

isotherm. Therefore the data for sorbed P were

adjusted by adding Olsen-P to estimate pre-existing
labile P, and then fitted to the Freundlich isotherm:
(mmol kg-1 pH-1)

P ¼ a Cb ð1Þ
pH buffering

The product of the Freundlich parameters, ab, was

also used in the regression analyses because at a P
12.68
20.40
72.50
32.73
12.59

12.84

18.02
10.73
24.33
37.03
7.51

8.85

concentration of 1, a equals the amount of P sorbed,

and ab equals the P buffer capacity at the same
pH, water

concentration. Phosphorus sorption properties of the

soils are given in Table 2.
7.9
6.1
6.8
8.3
7.4
5.0
5.5
5.0
5.4
5.1
8.8
3.4

Field capacity for water was estimated by flooding

soil with water and allowing it to drain freely for
Organic C

24 h. Experimentation showed that for most soils,

(g kg-1)

8.8
12.0
23.2

14.4
13.6

17.0
11.7
37.3
12.7
21.7
30.0
170.2

wetting air-dry soil to 60% of the field capacity gave

soil that was moist to touch and the texture in a
reasonable state if plants were to have been grown.
(mg kg-1)

Pwllpeiran, Neuhaus and Florasca required wetting to

Olsen P
Table 1 Characteristics of the soils as sampled

75% field capacity. Water contents were measured by

5

8
68
52
92
20
14

75

17
17
18

oven-drying overnight at 105°C. All results are

presented on an oven-dry basis.
Clay

8.3
2.4

4.7
18.8

78.7
26.4
25.9

46.1
40.2
31.6
53.0
16.1
(%)

Incubation procedure
Netherlands

Cameroon
Germany
Hungary

Hungary
Country

Zambia

Air dry soil (300 g) was mixed with the P amend-

Kenya
Israel
Italy

UK
UK

UK

ment, if used, water added to a water content of 60 or

70% field capacity (range 35–70% moisture content)
Whiteknights

and mixed again. The treatments were Control (no P),

Droevendaal

Rosemaund

Compost
Pwllpeiran

ABC, GPR and TSP, with additions at rates providing

Florascaa

Misamfu
Neuhaus
Albenga

Szentes
Kiboko

500 mg P kg-1 soil. There were three replicates of

Hula

Odja
Soil

each treatment. The soils were incubated at 20°C in

a

123
170 Nutr Cycl Agroecosyst (2009) 84:167–178

Table 2 Phosphorus
Soil P sorption index (cg kg-1) Freundlich isotherm parameters
sorption properties
-1
of the soils P addition rate (mg kg ) a (mg kg-1) b (l kg-1)
300 1500

Albenga 2.56 4.8 61.9 0.233

Droevendaal 8.29 10.9 125.3 0.325
Florasca 19.97 29.6 194.6 0.537
Hula 12.21 25.3 193.2 0.372
Kiboko 4.37 6.2 42.0 0.274
Misamfu 7.84 13.9 106.6 0.313
Neuhaus 2.99 5.2 67.9 0.237
Odja 8.68 12.6 97.4 0.326
Pwllpeiran 18.74 27.3 264.8 0.370
Rosemaund 5.82 9.1 66.6 0.352
Szentes 8.02 14.2 96.4 0.397
Whiteknights 14.64 23.6 205.4 0.352

the dark in polyethylene containers with ventilation
holes. Soil water content was maintained by occa-
sional addition of purified water to weight. At days 1,
5, 13, 34, 70 and 145, soil samples were taken from
each replicate for Olsen P and P dissolution analysis.
For some soils, there were minor deviations from this
sampling plan to fit with the calendar. At the end of
the incubation period, a portion of the remaining soil
was air-dried at 30°C and ground to pass a 2-mm
aperture sieve prior to storage for additional analysis.
Dissolved-P in soil
A two-step sequential extraction was performed
following the method of Mackay and Syers (1986).
Soil (containing 0.8 g oven-dry mass) was weighed
into a pre-weighed polymer centrifuge tube (50 ml).
NaCl solution (1.0 M, 40 ml) was added and the tube
shaken end-over-end for 30 min. Following centrifu-
gation (RCF = 500g), the supernatant was carefully
decanted and the tube with moist soil reweighed.
NaOH solution (0.5 M, 40 ml) was added and the
tube shaken with a vortex mixer to re-disperse the
soil. The sample was shaken end-over-end for 16 h,
centrifuged (RCF = 2,800g) and the supernatant
filtered (Whatman 42 filter).
P in the NaCl extract was measured colorimetri-
cally by the molybdenum blue and ascorbic acid
method (Kuo 1996). An aliquot of the NaOH extract
was neutralized with 1 M H2SO4, centrifuged
(RCF = 2,800g) to remove precipitated humic acid,
and the P concentration measured colorimetrically. In
calculation of the results for P extracted by NaOH,
allowance was made for P in the NaCl solution that
was entrained in the moist soil immediately before
addition of NaOH. Dissolved-P was defined as the
difference between treated soil and Control soil (the
mean value) for the P extracted by NaOH.
Description of dissolution rates for ABC
The following equations with two fitted parameters
are commonly used to describe the kinetics of soil
processes such as sorption and dissolution (Sparks
1989):


First order: Q ¼ a 1  ebT ð2Þ
Elovich: Q ¼ a þ b ln (T) ð3Þ
Powerequation: Q ¼ a Tb ð4Þ
where Q is the quantity of product at time T in days,
and a and b are fitted parameters. In presentation of
results, a and b are re-used in the different equations,
but this does not imply any relationship or connection
between the same parameter symbol used in different
equations. Dissolved-P was used for Q. The results
were fitted by linear regression for the Elovich
equation and by non-linear regression for First order
and Power equations.

123
Nutr Cycl Agroecosyst (2009) 84:167–178 171

Statistics Olsen-P in soil

All statistical calculations were made with Genstat The addition of ABC produced an immediate signif-
(Lawes Agricultural Trust 2006). icant increase in Olsen-P after one day of incubation
in all soils except Albenga and Szentes (Table 4).
The increase, relative to the control, ranged from
Results 6.1 mg kg-1 (Hula) to 44.9 mg kg-1 (Whiteknights).
In most soils where ABC caused immediate Olsen-
ABC and Gafsa PR P increase, Olsen-P increased further over the incu-
bation period, and by the end of incubation the value
The ABC and GPR contained similar concentrations was changing little. In Szentes soil with ABC, Olsen-
of total P (13.4 and 12.5%, respectively). However, P remained unchanged, while in Albenga, Florasca
ABC contained very much more P in all extractable and Hula, Olsen-P declined (Table 4). In Florasca
forms; for example, 244 mg kg-1 water extractable P and Hula soils, ABC initially gave an increase in
compared with 42 mg kg-1 in GPR (Table 3). Olsen-P, but not after 145 days.
Organic C was a minor component (10.4%) of ABC. GPR gave an immediate increase in Olsen-P only
in Odja, Neuhaus, Misamfu and Whiteknights soils.
In the same four soils plus Pwllpeiran, there was a
significant increase in Olsen-P during incubation. In
Table 3 Properties of ABC and Gafsa PR
Albenga and Florasca, there were decreases in Olsen-
ABC GPR P during incubation with GPR, in parallel with those
Total P (%) 13.4 12.5 seen in those soils incubated with ABC.
Neutral ammonium citrate extractable P (%) 11.5 4.1 TSP caused a significant initial increase in Olsen-P
Olsen P (mg kg ) -1
1498 116 for all soils (Table 4), but the size of the increase
Water extractable P (mg kg-1) 244 42
differed between soils and for all TSP treated soils,
Total C (%) 12.50 2.13
Olsen-P was much less than the Control value plus
Organic C (%) 10.4 0.38
500 mg P kg-1. During incubation, Olsen-P declined
in all soils except Szentes and Whiteknights. This
Surface area (m2 g-1) 82.2 16.5
suggests that a substantial and variable portion of the
pH (water) 7.6 6.4
soluble P was sorbed in all soils. The net sorption was

Table 4 Olsen-P (mg kg-1) in soils after incubation without P and with ABC, GPR and TSP
Soil Day 1 Day 145
Control ABC GPR TSP Control ABC GPR TSP

Albenga 54.6 55.6 53.2 429.0 46.6 44.7 41.7 240.1

Droevendaal 52.1 63.8 52.0 235.5 50.2 68.7 50.7 160.1
Florasca 55.2 62.3 58.6 234.7 39.2 41.2 38.9 122.7
Hula 14.2 20.3 15.1 303.3 13.4 14.9 13.3 73.7
Kiboko 13.8 20.5 13.8 432.9 13.9 25.6 12.7 239.0
Misamfu 5.1 48.2 15.0 323.6 5.3 70.8 36.6 167.3
Neuhaus 68.4 88.5 72.1 329.0 67.7 127.8 93.1 271.1
Odja 1.5 25.0 7.2 179.9 1.4 65.0 34.8 133.5
Pwllpeiran 14.2 30.9 16.0 193.8 12.1 47.1 26.5 66.5
Rosemaund 15.0 26.9 15.6 304.9 12.8 44.0 19.1 148.2
Szentes 13.9 16.9 13.8 285.5 11.0 13.1 11.4 287.8
Whiteknights 7.5 52.4 44.6 168.2 11.2 146.7 118.1 174.0
s.e. (df = 71) 1.223 1.223 1.223 62.7 1.611 1.611 1.611 22.77

123
172 Nutr Cycl Agroecosyst (2009) 84:167–178

Power
300
y = -0.6847x + 236.79

51.6
10.0
87.4
36.3
81.8
88.6
81.6
87.8
250 R 2 = 0.51

Table 5 Fitted parameters of the First order, Elovich and Power equations, used to describe Dissolved-P from ABC (mg kg-1) in relation to time (days) for eight soils
Olsen-P from TSP,

% Variance accounted for

200
mg/kg

Elovich
150

50.1
10.8
86.2
39.7
84.3
84.1
81.9
84.8
100

50

First order
0

29.0
23.6
42.7
39.3
73.3
80.5
60.0
55.0
0 100 200 300
Freundlich parameter a

0.0422
0.0577
0.0166
0.0616
0.0423
0.0332
0.0254
0.0214
Fig. 1 Olsen-P derived from TSP applied at 500 mg P kg-1, at

s.e.
Day 145 of incubation, in relation to Freundlich parameter a of
the soil P sorption isotherms

Power equation parameters

0.1702
0.0899
0.1660
0.1740
0.2973
0.3119
0.1963
0.2117
estimated by the difference between TSP and Control

b
treatments for Olsen-P. Hula and Pwllpeiran soils

5.12
4.70
5.65
9.56

7.92
3.37
10.7

10.3
showed the greatest TSP sorption. It was correlated

s.e.
significantly with P sorption, assessed by the two P
sorption indices, and highly significantly with Fre-

31.0
22.2
87.5
39.9
60.4
56.7
33.6
118.9
undlich sorption isotherm parameter a (Fig. 1).

1.90
1.53
2.19
3.47
4.56
4.42
1.29
4.54
ABC dissolution rate
s.e.

Data for Dissolved-P from ABC were successfully 8.19

2.63
22.70
12.10
43.77
42.16
11.40
44.42
fitted to the First order, Elovich and Power equations
b

for eight soils (Table 5). For Kiboko soil, there was
Elovich parameters

significant dissolution averaged over all times and

6.40
5.08
7.19

4.22
11.4
14.9
14.5

14.9
s.e.

thus in parameter a, but the change from Day 1 to 145

and thus the curvature of the relationship between
29.0
21.5
81.0
33.5
31.7
35.6
29.3
104.2
Dissolved-P and time was not significant. For the
remaining four soils, there was no significant ABC
a

dissolution. For most soils, the Elovich and Power

0.0746
0.0852
0.0189
0.0114
0.0505
0.0380

equation fitted almost equally well, judged by

0.186
0.414
s.e.

variance accounted for (Table 5), but since the Power

equation gave the best fit for more soils than any
0.403
0.782
0.261
0.197
0.072
0.054
0.192
0.144

other equation, it was used to describe the dissolution

First order parameters

rate in subsequent work.

b

4.73
2.77
9.89
7.87

4.35
16.5
15.1

18.8
s.e.

Proportion of dissolved-P and soil pH

Dissolved-P was expressed as a percentage of the

59.3
31.7

83.6

75.6
170.5

229.2
240.3

292.8

amount of P added, and abbreviated to PD34 and

a

PD145, for 34 and 145 days’ incubation respectively.

Whiteknights

In five soils, PD145 was not significantly different

Droevendaal

Rosemaund
Pwllpeiran

from zero (Table 6). The highest soil pH for signif-

Misamfu
Neuhaus
Kiboko

icant P dissolution from ABC was 6.14 (Droevendaal

Odja
Soil

soil).

123
Nutr Cycl Agroecosyst (2009) 84:167–178 173

Table 6 Dissolved-P from ABC, GPR and TSP after 145 days’ incubation, as a percentage of P applied (PD145)
Soil PD145 Effect of P source on soil pH (water)
ABC GPR TSP ABC GPR TSP

Florasca -2.8 -4.1 55.7 -0.06 0.06 -0.51

Albenga -1.9 -0.8 -0.4 0.00 0.09 -0.74
Hula -0.3 -0.0 5.9 -0.03 0.02 -0.50
Szentes 0.1 -0.1 56.1 -0.05 0.11 -0.86
Kiboko 8.0 0.6 60.2 0.07 0.11 -0.56
Droevendaal 16.5 1.3 89.8 0.24 0.19 -0.11
Neuhaus 16.9 6.5 56.5 0.43 0.23 -0.29
Rosemaund 19.4 4.6 85.1 0.59 0.34 -0.22
Misamfu 39.6 20.6 82.4 0.98 0.83 0.21
Odja 49.3 28.2 88.8 0.26 0.29 0.10
Pwllpeiran 54.1 23.3 93.7 1.10 0.96 0.19
Whiteknights 68.0 52.5 80.1 0.32 0.46 0.02
Soils are placed in rank order of increasing PD145 for ABC. The effect of P source on soil pH was measured on soils after incubation.
The SEM (df = 72) was 3.00 for PD145 and 0.081 for effect of P source on soil pH

For GPR, PD145 was significant in only four soils
(Table 6). Compared with ABC, a lower pH was
required for P dissolution, less than 5.03 (Misamfu
soil). Both ABC and GPR caused significant
increases in pH for seven of the soils (Table 6).
These soils were the same seven where PD145 for
ABC was significant. TSP normally lowered pH.
The relationships of PD34 and PD145 with pH
appeared curved because there are four soils with high
pH values and non-significant P dissolution, while at
soil pH less than about 7.0, the relationship between pH
and PD145 was much closer to linear (Fig. 2). It was
considered that if P dissolution did not occur above
about pH 7, then the weaker influence of other soil
80
70
60
50
PD145 (%)
properties could be masked for those soils of pH \7.
Therefore, the correlations to assess factors influencing
ABC dissolution were based only on the eight soils
with measurable P dissolution from ABC, comprising
the seven with significant PD145, plus Kiboko.
Although the PD145 for Kiboko soil was not signif-
icant, the fitted dissolution curves did demonstrate
significant P dissolution on average (rate parameter a).
Correlations between ABC dissolution and soil
properties
Significant inverse correlations were found between
PD34 and PD145, and soil pH (Table 7). Significant
positive correlations were found between PD34 and
PD145, and the following properties relating to P
sorption: Freundlich parameters a and ab, and P
sorption index at both rates of P addition. Fitted
parameter a of the Power equation for dissolution rate

40 correlated significantly with soil pH, but not P

30 pH KCl sorption properties, while parameter k showed no
20 pH water correlation with soil properties. However, dissolution
10 rate parameter a was significantly correlated with
0 exchangeable acidity and exchangeable Al.
-10 2 4 6 8 10 In most soils, PD145 was greater than PD34, but
soil pH the incidence and closeness of significant correlations
with soil parameters was the same for these two
Fig. 2 Relationships between ABC dissolved at 145 days
(PD145) and initial soil pH in 12 soils. Fitted quadratic lines assessments of P dissolution (Table 7). Therefore, it
are shown for pH in water (dashed) and KCl (solid) was decided to use PD145 as the principal assessment

123
174 Nutr Cycl Agroecosyst (2009) 84:167–178

Table 7 Extract from the matrix of correlation coefficients parameters a and k for dissolution of ABC, and soil properties,
between dissolved-P at 34 and 145 days (PD34 and PD145 for the eight soils with measurable P dissolution from ABC
respectively) for ABC, PD145 for GPR, power equation
GPR ABC ABC
PD145 PD34 PD145 Power Eq-a Power Eq-k

PD34 (ABC) 0.955*** 1

PD145 (ABC) 0.957*** 0.957*** 1
Power Eq-a 0.930*** 0.890** 0.866** 1
Power Eq-k 0.675 0.675 0.707* 0.309 1
Olsen P -0.508 -0.494 -0.536 -0.456 -0.263
Pindex-300 0.673 0.801* 0.816* 0.549 0.693
Pindex-1500 0.746* 0.864** 0.869** 0.657 0.664
Freundlich-a 0.638 0.776* 0.776* 0.544 0.656
Freundlich-b 0.484 0.583 0.626 0.389 0.612
Freundlich-ab 0.625 0.766* 0.770* 0.52 0.667
pH, water -0.835** -0.823* -0.801* -0.846** -0.510
pH, CaCl2 -0.807* -0.786* -0.759* -0.822* -0.501
pH, KCl -0.888** -0.872** -0.845** -0.924** -0.480
pH Buffer capacity 0.617 0.517 0.510 0.548 0.085
Exchangeable acidity 0.844** 0.704 0.685 0.835** 0.083
CEC -0.252 -0.286 -0.268 -0.359 -0.139
Exchangeable Al 0.818* 0.670 0.649 0.807* 0.055
Exchangeable Ca -0.523 -0.498 -0.476 -0.613 -0.145
Exchangeable K -0.382 -0.506 -0.494 -0.413 -0.641
Exchangeable Mg -0.355 -0.420 -0.378 -0.486 -0.261
Exchangeable Mn 0.182 0.239 0.302 -0.005 0.661
Exchangeable Na -0.262 -0.180 -0.136 -0.449 0.153
Clay 0.215 0.322 0.377 0.037 0.602
Organic C 0.408 0.507 0.503 0.262 0.463
Total C 0.420 0.526 0.524 0.269 0.498
Total P -0.192 0.016 0.043 -0.382 0.461
*, ** and *** denote correlations significant at the 5, 1 and 0.1% levels respectively

of P dissolution because it related to more soil

properties than dissolution rate parameter a, and
being a larger value than PD34 was considered to be
a more reliable measurement.
Multiple correlations of PD145 with soil properties
were investigated and the best was with pH (KCl) and
Pindex-300 (% variance accounted for = 91.9%;
Fig. 3):
PD145 ¼72:1  12:2  pHðKClÞ
ð5Þ
þ 2:22  Pindex-300
An alternative multiple regression with a good fit
existed with pH in water and Pindex-1500 (%
variance accounted for = 84.5%):
PD145 ¼57:6  8:62  pHðwaterÞ
ð6Þ
þ 1:70  Pindex-1500
Correlations between GPR dissolution and soil
properties
Significant correlations were found between PD145
for GPR and soil pH, exchangeable acidity and Al,
and Pindex-1500 (Table 7). PD145 for ABC
(PD145[ABC]) and for GPR (PD145[GPR]) were
very closely correlated (r = 0.957, Table 7),
although PD145[GPR] was consistently lower, as
shown by the regression equation:

123
Nutr Cycl Agroecosyst (2009) 84:167–178 175

80 exists as hydroxyapatite and that dissolution was to

70 H2PO4-, then the dissolution equation is:
60
Ca5 ðPO4 Þ3 OH þ 7Hþ ! 5Ca2þ þ 3H2 PO4  þ H2 O
PD145 (%)

50
40 ð8Þ
30 It shows that dissolution of 3 mol of P require 7
20 mol of acidity (H?); therefore 500 mg kg-1 of P
10 would require (7/3) 9 (500/30.97) = 37.7 mmol kg-1.
0 PD145 shows that 68% of ABC was dissolved
0 20 40 60 80
(Table 6), thus consuming 25.6 mmol kg-1 acidity,
Fitted Value
while the measured consumption was 32.2 mmol
Fig. 3 Multiple regression equation describing PD145 (ABC kg-1. Given that there is substitution of both cations
dissolved at 145 days) in relation to soil pH and soil P sorption and anions in the ABC, and other components such as
index, for eight soils with significant P dissolution. Fitted carbonate that may consume acidity, this is considered
value = 72.1 - 12.2 9 pH(KCl) ? 2.22 9 Pindex-300
to be reasonable agreement. Thus the acidity required
to dissolve ABC was provided largely by the inorganic
PD145½GPR ¼ 0:781  PD145½ABC  9:34 ð7Þ exchangeable acidity.

Exchangeable acidity in Whiteknights soil

Discussion
Exchangeable acidity was measured in the dried
Influence of soil properties on ABC dissolution
Whiteknights soil after incubation. This soil was
analysed because it gave the highest PD145 and
The data for dissolution rate of ABC were interpreted
should therefore demonstrate most clearly the
by fitting to three Eqs. 2–4 that have been commonly
demands made on soil acidity for P dissolution.
used to assess dissolution rates of materials added to
Dissolution of GPR and ABC consumed a large part
soil. For most soils, the Power Function (4) gave the
of the exchangeable acidity (Table 8). Estimates for
best fit, assessed by the % variance accounted for
the actual and theoretical consumption of acidity on
(Table 5). Equations 2 and 3 may be justified by
dissolution of ABC were compared. The difference in
theoretical mechanisms for dissolution reactions
total exchangeable acidity between ABC and control
(Chien et al. 1980a). However, although the relation-
treatments was 32.2 mmol kg-1. This difference was
ship is empirical, equations of the form of (4) in
caused by consumption of acidity in order to dissolve
respect of time describe well the time course of
ABC in Whiteknights soil. Whiteknights soil pH after
desorption of phosphate from soil and soil compo-
incubation with ABC was 3.83, and pKa values for
nents (Barrow 1979).
steps 1 and 2 of orthophosphoric acid are 2.12 and
Equation 4 fitted the data best for the eight soils
7.21, so it is expected that H2PO4- is the dissolution
in which significant P dissolution took place.
product at pH 3.83. If it is assumed that P in ABC
Therefore, its fitted parameters were the logical
ones to correlate with soil properties and parameter
Table 8 Exchangeable acidity (cmol kg-1) in Whiteknights a showed significant correlations with soil properties
soil after 145 days’ incubation
related to acidity (Table 7). However, for this
Treatment Al acidity H acidity Total acidity purpose of investigating the influence of soil prop-
erties on ABC dissolution, the simple percentages of
Control 4.34 0.70 5.05
Dissolved-P from ABC (PD34 or PD145) were
ABC 1.43 0.40 1.83
preferred, being correlated with more soil properties,
GPR 1.74 0.36 2.11
and would be much easier to measure in future
TSP 3.12 0.66 3.78
work, since one measurement only is needed instead
SED (df = 10) 0.089 0.063 0.073
of a time series.

123
176
Soil acidity was clearly a major constraint on P
dissolution from ABC. This is shown by (i) the
significant correlations of PD145 with soil pH
(Table 7), (ii) the increases in soil pH when P
dissolution occurred from ABC (Table 6), (iii) the
significant correlation of dissolution rate parameter a
with exchangeable acidity and Al, and (iv) the
consumption of exchangeable acidity in Whiteknights
soil (Table 8).
Dissolution of calcium phosphates such as ABC
releases Ca and H2PO4- into soil solution (Eq. 8),
and therefore the reaction will not proceed if soluble
Ca and H2PO4- concentrations rise too far. A sink for
P dissolved was also an important soil property that
influenced P dissolution from ABC. This was shown
by significant correlations of PD34 and PD145 with
the two P sorption indices and the ‘‘extensive’’
parameter a of the P sorption isotherms (Table 7).
The latter relates to the P sorption capacity of the soil
(Holford 1982), whereas the parameter b is an affinity
parameter (Holford 1982) for P sorption, and was not
correlated with the extent of ABC dissolution, again
suggesting that the capacity of the soil to sorb
released P is an important influence. Dissolution of
PR has been shown to be positively affected by the
size of Ca sink (Robinson et al. 1992), but there
was no significant correlation of PD145 with CEC
or exchangeable Ca, either alone or in multiple
regressions. This suggests that the sink for Ca was not
an important factor in assisting P dissolution from
ABC.
After addition of 500 mg kg-1 soluble P as TSP,
the P remaining extractable as Olsen-P was lower in
soils with higher P sorption as expected (Fig. 1). This
shows that soluble P released by dissolution from
ABC is sorbed in the soil, reducing its plant
availability, but providing a bigger sink and therefore
encouraging P dissolution. The complementary
nature of the P sink with acidity is illustrated by the
very close multiple correlation of the two with PD145
(Eqs. 5, 6).
The two multiple correlations of PD145 with pH
and P sorption (Eqs. 5, 6) suggest that relationships
could be obtained to predict the likely extent of ABC
dissolution in a soil from its known properties.
Equation 5 gave the closer correlation, but Eq. 6 uses
variants of the measurements that are more com-
monly in routine use for other purposes, and so are
better suited for predictive use.
123
Nutr Cycl Agroecosyst (2009) 84:167–178
Comparison of P dissolution from ABC and GPR
In general, initial soil pH is a good predictor of PR
dissolution (Rajan et al. 1996), and we found the
same applied to dissolution of ABC. An approximate
cut-off pH value could be identified above which P
dissolution was insignificant, for GPR ([5.03) and
ABC ([6.14). For both ABC and GPR, the closest
correlations with soil properties were with pH, while
the correlation of GPR dissolution was less close with
P sorption (Table 7). For the eight soils with signif-
icant ABC dissolution, the relationship between soil
pH and PD145 approximated to linear below pH 7
(Fig. 2), and other linear relationships between soil
pH and PR dissolution have been found, e.g., in acid
New Zealand soil (Bolan and Hedley 1990). Corre-
lation of P dissolution parameters for North Carolina
PR in 16 Colombian acid soils showed significant
effects of P sorption, pH and exchangeable Al (Chien
et al. 1980b). The same effects were found in this
work with GPR, but Chien et al. (1980b) found
additional correlations with clay and organic matter,
which were not found in our group of soils. However,
we can conclude that the generally established
controlling principles for P dissolution from PR also
apply to ABC.
Agronomic effectiveness of ABC
Recent data on the availability of P to plants from
modern recycling products are rare (Romer 2006).
Studies show that products such as MBM (meat and
bone meal), steamed bone meal and bone chips give P
responses to pot and field grown crops (Romer 2006;
Jeng et al. 2006; Klock and Taber 1996). ABC is
manufactured using much higher temperatures (ca.
400°C) than the previously mentioned materials and
contains less organic matter. It was therefore
expected to have different surface properties and
different dynamics of dissolution and availability
when compared with PR and the above-mentioned
materials.
Olsen-P is a widely accepted laboratory index of
the plant-available P in soil, and it is useful in both
acid and calcareous soils (Kuo 1996), a particular
advantage in the present work where a wide soil pH
range was intended. At the end of the incubation
period, the soils where significant P dissolution from
ABC occurred (Table 6) were the same ones that
Nutr Cycl Agroecosyst (2009) 84:167–178
gave significant increases in Olsen-P (Table 4),
showing that P dissolution is a guide to the P
availability to plants and that ABC will be an
effective P fertilizer in many soils.
When P dissolution occurred, the majority took
place in the first 20 days. Because the mobility of P in
soil is low, the early availability of P is essential for
proper crop development (Hedley et al. 1995). ABC
showed a clear benefit to Olsen-P over GPR for most
soils (Tables 4, 5). In two soils, Florasca and Hula,
ABC improved Olsen-P at Day 1 although not at Day
145, and these soils were high pH soils where GPR
was completely insoluble. Thus, for most soils, ABC
provides a limited but immediately available P pool
in soil, combined with further P release thereafter, a
pattern of release that appears well timed to suit crop
requirements. This pattern is different to the one for
TSP, where Olsen-P starts at a much higher concen-
tration than with ABC and then declines because of
the reaction of dissolved phosphate with soil
(Table 4). However, it should be noted that for
slowly-soluble fertilizers, Olsen-P gives a measure of
the availability of immediate reaction products only
and underestimates plant-available P (Menon and
Chien 1995; Saggar et al. 1992). Thus in comparison
with TSP, plant-available P from ABC may be better
than our data suggest. It is clear that ABC is a
fertilizer with P supply characteristics intermediate
between traditional slow-release (PR) and fully
soluble (TSP) fertilizers. The results call for pot and
field trials to test this conclusion.
Conclusions
The soil properties that influenced P dissolution from
ABC were the same as those that control P dissolu-
tion from PR, as originally hypothesised. In
particular, the most important soil properties were
those related to soil acidity, and the more exchange-
able acidity and lower the pH, the greater the
dissolution of ABC. The size of the sink for
Dissolved-P was the other significant control on
ABC dissolution, and the two could be combined in
multiple regression Eqs. 5 and 6 that use readily
measured soil properties to predict the potential for
ABC dissolution in a soil. This conclusion is based on
a correlation study with a wide range of soils, and
calls for detailed mechanistic studies on the
177
mineralogical and surface properties of ABC and P
dissolution from it.
The above conclusion implies that the knowledge
of the soils and circumstances when PR is useful as a
fertilizer applies also to ABC. However, in a direct
comparison with GPR, which is well-characterised
and widely accepted as a relatively soluble PR, ABC
was much better in providing available P, as assessed
by the Olsen method. It could even supply some
available P immediately after application to soils of
high pH, in which GPR was ineffective. Therefore,
ABC is predicted to have a wider range of application
than PR, so that the soils for which it is suitable and
rates of ABC addition need to be assessed under field
conditions. Because of its apparent potential as a P
fertilizer, field trials are now in progress.
Acknowledgements This work was financially supported by
the European Union, Framework VI (Contract no. FOOD-CT-
2005-514082). We thank the anonymous reviewers for their
helpful comments.
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Smith AM (1959) Manures and fertilisers. Nelson, London
Sparks DL (1989) Kinetics of soil chemical processes.
Academic Press, London
Sumner ME, Miller WP (1996) Cation exchange capacity and
exchange coefficients. In: Sparks DL (ed) Methods of soil
analysis Part 3 chemical methods. Soil Science Society of
America and American Society of Agronomy, Madison,
pp 1201–1229
Van Kauwenbergh SJ (1997) Cadmium and other minor ele-
ments in world resources of phosphate rock. In:
Proceedings No. 400 International Fertiliser Society,
York, UK pp 1–40
 Oil Mallee charcoal properties
CEC meq/100g

volatiles%dry

ash%dry

fixed C%wet

fixed C%dry

s.area m2/g

0 20 40 60 80 100
%

pH(CaCl2) 8.4 EC 25 uS/m N 1.2% P 0.12% K 0.7% Ca 2.8% Zn 38 ppm

deep banding charcoal at
Pindar April 2005; Ausplow
Deep Blade System.
thanks to John Ryan
 DEEP BANDED MALLEE CHARCOAL

1 t/ha at broad-acre rate

for row spacing of 600 mm visible effect at
6 t/ha rate

6 t/ha in 100 mm wide band

 TRIAL SITES

PINDAR; sandy clay loam KALANNIE; yellow sand

40ppm av. P 44ppm av. P
 Pindar Kalannie 2 Kalannie 1
2500

55 kg/ha soluble fertiliser - May sown

2000
grain yield, kg/ha

1500

110 kg/ha soluble fertiliser - June sown

1000

500

0
0 1 2 3 4 5 6 7
rate of deep banded charcoal, t/ha

poor yield increase with recommended rates of soluble fertiliser

 half rate full rate
2500

+340 kg/ha
2000
grain yield, kg/ha

1500

Pindar
1000

500

0
0 1 2 3 4 5 6 7
rate of deep banded charcoal, t/ha

18% yield increase with half rates of soluble fertiliser at 6 t/ha char
 mineral plus microbes

2500

2000
+640 kg/ha
grain yield, kg/ha

1500

1000

500

0
0 1 2 3 4 5 6 7
rate of deep banded charcoal, t/ha

46% yield increase with at least 1.5 t/ha char added to the mineral fertiliser
 values of yield increases
yield banded broad acre value ($/ ha) for carbon
Trial site# benefit charcoal equivalent wheat at value
and fertiliser kg/ha kg/ha
kg/ha t/ha charcoal carbon $150/t $250/t $/kg C#
1. 100 kg/ha
mineral+1 640 1.5 250 56 96 160 2.84
1. 30 kg/ha
soluble 2 344 6.0 1000 225 52 86 0.38
2. 110 kg/ha
soluble 3 76 6.0 1000 225 11 19 0.08
3. 110 kg/ha
soluble* 83 3.0 620* 140 12 21 0.15
#
wheat at $250/t

~$3/kg = $3000/t of carbon!

 Mineral+ half rate of soluble full rate of soluble
2.6

2.4

2.2
Pindar
grain yield, t/ha

2.0

1.8

1.6

1.4

1.2

1.0
10 15 20 25 30 35 40
grains per head

grains/ head explained 42% of yield variation = drought stress effect

(the crop needed an ‘irrigation’ to survive 2005)
 55 kg/ha Sol. 30 kg/ha Sol. Min+

50

45
Pindar
root colonisation; flowering, %

40
at flowering
35

30

25

20

15

10

0
0 1 2 3 4 5 6 7
deep banded charcoal, t/ha

char increased AMF colonisation (especially the inoculated microbes)

 HS LS M+
35

3
30 6
0

6
Grains per head

25
0 3
6 3

20 Pindar
0
at flowering
15

10
0 5 10 15 20 25 30 35 40 45 50
root colonisation September %

AMF colonisation associated with grains/head

- may have helped reduce drought stress – fungal hyphae extend root system
 S55 S30 M+ S110

30

25
SYMBIOTIC ?
2
biomass at tillering, g/m

20

15
PARASITIC ?

10

0
0 1 2 3 4 5 6 7
banded charcoal, t/ha

char increased early growth for lower soluble P conditions

But decreased early growth for higher soluble P conditions
 Interpretations
• Valuable yield increases from char addiction
– broadacre agriculture, low native AMF
• Very efficient Carbon sequestration with low
rates of low C char
– Few soil effects, long term benefits?
• Char seems to have increased drought
tolerance by encouraging symbiotic fungi.
• Higher levels of soluble P may suppress
symbiosis – more value in low fertility situations.
 Improving wheat production with deep
banded Oil Mallee Charcoal in Western
Australia
Paul Blackwell , Syd Shea , Paul Storer , Zakaria Solaiman ,
1 2 3 4

Mike Kerkmans5, and Ian Stanley6

1Department of Agriculture and Food, Geraldton WA
2 Oil Mallee Company of Australia

3Western Mineral Fertilisers

4University of Western Australia

5Oil Mallee Association of WA

6 "Bungadale", Kalannie , WA
 location
• Mostly winter rain
WESTERN AUSTRALIA from May to October

• Many sandy soils

with organic matter
GERALDTON <1%
Pindar
Kalannie •Usually grows
PERTH
> 50% of Australia’s
wheat
WHEAT BELT
 helped develop
Oil Mallee
20 years
concept with
R&D into
John Bartle of
charcoal
CALM
and soil
fungi in Director of Oil
Japan and Mallee company
Makoto Ogawa Syd Shea
Indonesia

ALLEY planting

MALLEE in WA BLOCK planting

potential
~10 Mt/yr biomass
from Ogawa, M. 1994.Symbiosis of
People and Nature in the Tropics.
Farming Japan Vol.. 28 – 5, p10 21.

CHARCOAL improves the MICRO-HABIAT of beneficial soil microbes

 Some new fertiliser
suppliers are using
beneficial soil microbes
and mineral fertilisers.
Is charcoal a better
source of
microporosity than
zeolite?
 Research questions
• Will charcoal improve crop yields from
poor soil in a dry Mediterranean
environment?
– (50% increase in Sumatra; Yamato et al.,
220% increase in Brazil; Lehmann & Rondon)
• How much soluble fertiliser
• Will mineral fertiliser and inoculated soil
microbes enable more yield with charcoal?
Soil biological nutrition model with
charcoal
charcoal
particle

plant
root

Symbiotic
fungi (AM)
mineral
particle
 THE OIL MALLEE INDUSTRY
processing Mallee
for eucalyptus oil

OIL DISTILLATION

HARVESTING
Pyrolysis with an open pan ‘Moki’ method
at Kalannie by Shea, Stanley and Okimori
March 2005; air temp. = 40oC+!
Yasuyuki Okimori
wood charcoal (Jarrah) mallee charcoal (‘05 trials)

Mallee biomass after oil

extraction
 Oil Mallee charcoal properties
CEC meq/100g

volatiles%dry

ash%dry

fixed C%wet

fixed C%dry

s.area m2/g

0 20 40 60 80 100
%

pH(CaCl2) 8.4 EC 25 uS/m N 1.2% P 0.12% K 0.7% Ca 2.8% Zn 38 ppm

deep banding charcoal at
Pindar April 2005; Ausplow
Deep Blade System.
thanks to John Ryan
 DEEP BANDED MALLEE CHARCOAL

1 t/ha at broad-acre rate

for row spacing of 600 mm visible effect at
6 t/ha rate

6 t/ha in 100 mm wide band

 TRIAL SITES

PINDAR; sandy clay loam KALANNIE; yellow sand

40ppm av. P 44ppm av. P
 Pindar Kalannie 2 Kalannie 1
2500

55 kg/ha soluble fertiliser - May sown

2000
grain yield, kg/ha

1500

110 kg/ha soluble fertiliser - June sown

1000

500

0
0 1 2 3 4 5 6 7
rate of deep banded charcoal, t/ha

poor yield increase with recommended rates of soluble fertiliser

 half rate full rate
2500

+340 kg/ha
2000
grain yield, kg/ha

1500

Pindar
1000

500

0
0 1 2 3 4 5 6 7
rate of deep banded charcoal, t/ha

18% yield increase with half rates of soluble fertiliser at 6 t/ha char
 mineral plus microbes

2500

2000
+640 kg/ha
grain yield, kg/ha

1500

1000

500

0
0 1 2 3 4 5 6 7
rate of deep banded charcoal, t/ha

46% yield increase with at least 1.5 t/ha char added to the mineral fertiliser
 values of yield increases
yield banded broad acre value ($/ ha) for carbon
Trial site# benefit charcoal equivalent wheat at value
and fertiliser kg/ha kg/ha
kg/ha t/ha charcoal carbon $150/t $250/t $/kg C#
1. 100 kg/ha
mineral+ 1 640 1.5 250 56 96 160 2.84
1. 30 kg/ha
soluble 2 344 6.0 1000 225 52 86 0.38
2. 110 kg/ha
soluble 3 76 6.0 1000 225 11 19 0.08
3. 110 kg/ha
soluble* 83 3.0 620* 140 12 21 0.15
#
wheat at $250/t

~$3/kg = $3000/t of carbon!

 Mineral+ half rate of soluble full rate of soluble
2.6

2.4

2.2
Pindar
grain yield, t/ha

2.0

1.8

1.6

1.4

1.2

1.0
10 15 20 25 30 35 40
grains per head

grains/ head explained 42% of yield variation = drought stress effect

(the crop needed an ‘irrigation’ to survive 2005)
 55 kg/ha Sol. 30 kg/ha Sol. Min+

50

45
Pindar
root colonisation; flowering, %

40
at flowering
35

30

25

20

15

10

0
0 1 2 3 4 5 6 7
deep banded charcoal, t/ha

char increased AMF colonisation (especially the inoculated microbes)

 HS LS M+
35

3
30 6
0

6
Grains per head

25
0 3
6 3

20 Pindar
0
at flowering
15

10
0 5 10 15 20 25 30 35 40 45 50
root colonisation September %

AMF colonisation associated with grains/head

- may have helped reduce drought stress – fungal hyphae extend root system
 S55 S30 M+ S110

30

25
SYMBIOTIC ?
2
biomass at tillering, g/m

20

15
PARASITIC ?

10

0
0 1 2 3 4 5 6 7
banded charcoal, t/ha

char increased early growth for lower soluble P conditions

But decreased early growth for higher soluble P conditions
 Interpretations
• Valuable yield increases from char addiction
– broadacre agriculture, low native AMF
• Very efficient Carbon sequestration with low
rates of low C char
– Few soil effects, long term benefits?
• Char seems to have increased drought
tolerance by encouraging symbiotic fungi.
• Higher levels of soluble P may suppress
symbiosis – more value in low fertility situations.
 Questions
• Can these results be repeated?
– pot trials UWA, small plot trials. NLP submission
• How low is the char requirement of the mineral
fertiliser?
– 2007 small plot trials
• How long can the char effect last?
– resowing Pindar and Kalannie trials
– NLP submission
– ARWA support a research potential seminar in June
– KEY information for potential Char economics
INTEGRATED WOOD PROCESSING (IWP) plant at NARROGIN

5MW plant = $6.2M over 20 years; est. Bell and Bennett (2002)
Ogawa and Okimori (2004)
Ogawa and Okimori (2004)
 A BENEFICIAL SYSTEM?

THANKYOU
Sylvain Pottier
Yasuyuki Okimori and Makoto Ogawa of
Kansai Environmental Engineering Centre,
Kansai Electric Co. Ltd and General
Environmental Technos Co., Ltd. and the
Oil Mallee Company for financial support
Ausplow Ltd for the use of their plot
airseeder. Andrew Donken
Victor Dodd and Doug Cail
Dave Gartner, Ben Parkin and Chris Gazey
United Farmers Cooperative and Hans
Schoof for soil testing and interpretation.
Stephen Davies, Bill Bowden, John Bartle,
and Tony Vyn for field advice and
assistance.
CSIRO PUBLISHING
www.publish.csiro.au/journals/ajea Australian Journal of Experimental Agriculture, 2007, 47, 1377–1382

Determining the agronomic value of composts produced

from garden organics from metropolitan areas of
New South Wales, Australia

K. Y. Chan A,B,E, C. Dorahy A,C and S. Tyler A,D

ACentre for Recycled Organics in Agriculture.
BNSW Department of Primary Industries, Locked Bag 4, Richmond, NSW 2753, Australia.
CNSW Department of Primary Industries, PMB 8, Camden, NSW 2570, Australia.
D432 Zara Road, Chillingham, NSW 2484, Australia.
ECorresponding author. Email: yin.chan@dpi.nsw.gov.au

Abstract. About 0.3 million t/year of composted garden organics (CGO) including mulches and soil conditioners are
produced annually in New South Wales, Australia, although only a small proportion of this material (<4%) is used in
agriculture. A lack of information on product characteristics and agronomic performance has limited the development of
agricultural markets for CGO products. These CGO products are the coarse and fine fractions separated by screening after
composting. This paper presents the results of a survey of CGO mulches and soil conditioners (unblended or blended with
a mixture of other organic materials including biosolids, animal manures and paper), which are commercially produced in
the metropolitan areas of New South Wales and assesses their agronomic and soil amendment values in terms of chemical
and biological properties. It also evaluates the short-term effects of applying increasing rates (0, 25, 50 and 100 t/ha) of
selected composted soil conditioners on radish growth in a pot experiment.
The mulch products had low nutrient concentrations but had high carbon (C) contents (mean C = 45%) and C/Nitrogen
(N) ratios (mean C/N = 72) and are most suitable for use as surface mulch. The unblended soil conditioners were low in
nutrients, particularly N (average total N = 1.0%, range 0.9–12%), and had lower and variable C contents. The pot trial
results indicated lack of growth response of radish at application rates up to 100 t/ha of unblended soil conditioners from
garden organics. The blended soil conditioners were more variable in quality and as confirmed by pot trial results produced
highly variable plant responses. The high variability in product quality and performance of the soil conditioners,
particularly the blended products might be related to the source and type of blending material as well as the composting
conditions used in the manufacturing process.
These results highlight the need to improve compost quality and consistency and the need for further research to
advance understanding of the benefits using CGO in terms of improving soil quality, crop productivity and net economic
returns to growers.

Introduction
Composts produced from plant and animal residuals have been
used by farmers worldwide for many hundreds of years to
increase crop productivity and improve soil quality but they are
not typically used as a component of Australian agricultural
production systems. However, Australian farmers are
increasingly seeking alternatives to conventional inputs to
provide plant nutrients and improve soil health as indicated by
the increasing interest in the use of organic fertilisers instead of
inorganic fertilisers.
Across Australia, government legislation (Resource NSW
2001; Victorian Consolidation Legislation 2005) and strategies
(Resource NSW 2003; Zero Waste SA 2005) focus on diverting
materials from landfills and the beneficial re-use of these
materials. For example, in New South Wales (NSW), Australia,
over 600000 tonnes of garden organics, such as grass clippings,
prunings and other vegetation, are collected from households
and municipal areas each year and composted to produce
~0.3 m t/year of composted garden organics (CGO) (DEC
2004). After composting, the product is usually screened into
coarse and fine fractions, to produce mulches and soil
conditioners, respectively.
Most (87%) of the recycled organics generated in the Sydney
metropolitan area are consumed in the urban amenity market
segments including landscaping, and domestic garden
applications (DEC 2004). However, the urban amenity market is
approaching saturation (WMAA 2005), whereas only ~4% of
the CGO are currently used in agriculture (DEC 2004). Given
the large area of arable agricultural land in NSW
(>100000 km2; ABS 2001) the size of the agricultural market is
virtually unlimited.
As a prerequisite for developing agricultural markets for
composts, there is a need to understand the characteristics of
CGO and demonstrate their agronomic value to farmers.
However, little information is available on the quality and
agronomic performance of the CGO currently being produced
in NSW.

© CSIRO 2007 10.1071/EA06128 0816-1089/07/111377

1378 Australian Journal of Experimental Agriculture
The objectives of this paper are: (i) to present the results of a
survey of CGO being commercially produced in the
metropolitan areas, particularly in the Sydney region; (ii) to
assess their agronomic and soil amendment values in terms of
physical, chemical and biological properties; and (iii) to
evaluate the short-term performance of selected soil
conditioners in a pot experiment.
Materials and methods
Survey of CGO
A survey of 13 composting facilities in the Sydney region and
other major metropolitan areas of NSW was conducted in 2004.
All the facilities surveyed employed open-windrow systems, the
dominant form of composting in most parts of Australia
(Wilkinson et al. 2002). At each facility, finished compost
products that were ready for market were collected. Composite
samples (10–20 kg) of composted soil conditioners and mulches
were obtained by shovelling 1–2 kg of compost from 10 random
positions in the standing pile into plastic bags. The composite
samples were thoroughly mixed and stored at 4°C before analysis.
In total, nine CGO mulches, 10 CGO soil conditioners and
12 blended CGO soil conditioners were collected and studied in
this research. Soil conditioners refer to the fine compost
fraction (<8–10 mm) produced from garden organics as the only
feedstock. Blended CGO soil conditioners are fine compost
fraction produced from either co-composting a mixture of
garden organics and other organic residuals, such as biosolids,
poultry litter and cattle feedlot manure or mixing CGO soil
conditioners with other organic residuals.
CGO characterisation
The samples of mulches and soil conditioners were analysed for
a range of chemical, physical and biological properties.
Immediately after sampling, moist samples were analysed for
pH, electrical conductivity, ammonium, nitrate and soluble
phosphorus (P) concentration following the procedures
described in Standards Australia (2003). The CGO soil
conditioners were also tested for phytotoxicity and microbial
activity. The phytotoxicity bioassay involved germinating radish
seeds on a sample of soil conditioner and assessing early root
growth against a minimum length requirement of 60 mm
(Standards Australia 2003). Microbial activity was measured
using the Food and Drug Administration (FDA) enzymatic test
(Schnurer and Rosswall 1982).
Subsamples of mulches and soil conditioners were dried and
ground (<2 mm) and analysed for total carbon (C), organic
nitrogen (N), total sulfur (S), total P, pH, electrical conductivity
(EC), total copper (Cu), total zinc (Zn) and total iron (Fe). pH
and EC were determined on 1 :5 sample/water extract following
Rayment and Higginson (1992). Total C, N and S were
determined using Leco combustion (Rayment and Higginson
1992). Total P, total Cu, total Zn, total boron and total Fe were
determined using an inductively coupled plasma mass
spectrometer after acid digestion (USEPA 1996).
Pot experiment
Four soil conditioners and six blended soil conditioners were
used in the pot experiment (Table 1). The blended soil
conditioners contained biosolids, poultry litter, cattle feedlot
K. Y. Chan et al.
manure and paper pulp and represented the range of materials
commonly used in compost manufacturing in NSW. All the
compost samples were brought to ~40% moisture content and
left at room temperature for 4 days before application.
The soil used for the pot trial was collected from 0–10 cm of
a grey Dermosol (Isbell 1996) located at the Centre for Recycled
Organic in Agriculture, near Camden (70 m Australia Height
Datum at 34.07536°S, 150.6956°E), NSW. The site had a long
history of cropping and cultivation but was under lucerne pasture
at the time of sampling. The soil properties were: pHCa (4.5);
organic C (12.9 g/kg); extractable P (21 mg/kg); water content
field capacity (–10 kPa) (0.28 kg/kg); clay content (260 g/kg).
Treatments and experimental design
Treatments involved 10 soil conditioners and four rates of soil
conditioner application, namely 0, 25, 50 and 100 t/ha on an
oven dry basis. The experiment followed a randomised complete
block design with four replications. Pots (115-mm internal
diameter and 100-mm tall) with individual trays were used. The
quantity of oven dry soil conditioners equated to 0, 25, 50 and
100 g/pot for the four application rates, respectively. 900 g of
oven dry equivalent of the screened soil was packed into each
pot. The appropriate amount of soil conditioner was then added
to the respective pot and thoroughly mixed. The individual pots
were then watered to field capacity (28% by weight) and left for
24 h in the glasshouse before sowing.
Ten radish seeds (Raphanus sativus L. cv. Long Scarlet) were
sown in each pot and these were thinned to five after plant
emergence. The pots were placed on four benches, each of
which represented an experimental block. All of the pots were
randomised within each block. During the experiment, all the
pots were maintained at approximately field capacity and
watered by hand on a regular basis.
After 26 days, the radish plants were harvested and the fresh
weight of both the tops and roots recorded. The harvested plants
were dried at 60°C to constant weight and re-weighed to
determine dry weight.
Statistical analyses
The different properties of the compost products were compared
using 1-way ANOVA in GENSTAT (VSN International Ltd 2003).
For the pot trial, results of both fresh weight and dry weight of
Table 1. Descriptions of the blended and unblended soil conditioners
derived from composted garden organics (CGO) used in the pot
experiment
Sample no. Type of conditioner Description
2 Soil conditioner CGO
3 Soil conditioner CGO
6 Soil conditioner CGO
28 Soil conditioner CGO
1 Blended soil conditioner CGO/biosolids
5 Blended soil conditioner CGO compost/poultry
litter/cow manure
11 Blended soil conditioner CGO/biosolids
14 Blended soil conditioner CGO/paper compost
16 Blended soil conditioner CGO/poultry litter
17 Blended soil conditioner CGO/poultry litter
Agronomic value of composted garden organics Australian Journal of Experimental Agriculture 1379

the radish using different soil conditioners and rate of
application were analysed using 2-way ANOVA. Differences
between treatments were statistically significant at 5%
probability unless otherwise stated. Product variability was
assessed by examining the ranges as well as coefficients of
variation (CV) of the different chemical parameters of the
different categories of composting products. Linear regression
analysis was also undertaken in GENSTAT to determine;
(i) whether radish dry weight was correlated with any of the
compost chemical parameters measured; and (ii) correlation, if
any, between results of FDA test and phytotoxicity bioassay.
Results
Product quality
Apart from size grading, the mulches had very different
properties compared with the soil conditioners (Table 2). The
mulches had lower total N, total P, total Ca and total magnesium
concentrations but higher C and C/N than the soil conditioners.
Mean C content of the mulches was 44.6%, which is nearly
twice that of the soil conditioners. Mean C/N of the mulches
was 71.7 compared with 25.5 of the soil conditioners.
Comparing the two categories of soil conditioners revealed
the blended soil conditioners had higher average EC, mineral N
and total S (Table 2) than the unblended soil conditioners. They
were also higher in heavy metals, as indicated by the Zn and Cu
concentrations in Table 2. Mean FDA values as well as CV were
similar between the two groups of soil conditioners (Table 3).
Similarly, mean root length, as a measure of phytotoxicity, was
not significantly different between the unblended CGO soil
conditioners and the blended products.
Product variability
While the C content of the mulches was fairly uniform (range
41–47%, CV = 4.5%), nutrient concentrations (total N and total
P) were more variable (CV >20%) (Table 2). Total N
concentration in the mulches varied more than 2-fold (range
0.5–1.0%), resulting in a similar magnitude of variation in the
C/N ratio (range 41–104). Higher variability was found in the
soluble/available nutrients, namely, soluble P and mineral N
(CV >50%).
Compared with the mulches, soil conditioners were more
variable in C content (15–32%, CV = 25.8%). While the C
content of the blended soil conditioners was similar to those of
the unblended soil conditioners, they were highly variable with
respect to most of the chemical parameters measured (Table 2).
For example, mineral N concentration ranged from 1.3 to
530 mg/kg (Table 2). The results from the phytotoxicity
bioassays (Table 3) showed that despite similar mean values, the
blended soil conditioners were more variable than the unblended
soil conditioners (CV 65.8 v. 13.7%; Table 3). In fact, 42% of
the blended products had root length <60 mm, the critical value
below which the medium is classified as phytotoxic (Standards
Australia 2003). In contrast, similar CV was found for the FDA
results of the two categories of soil conditioners.
Pot experiment
For the soil conditioners, no significant changes in fresh weight
of radish harvested 26 days after sowing with increasing
application rates up to 100 t/ha were observed (Fig. 1). In
contrast, radish fresh weight increased in response to increasing
application rate in five out of the six blended soil conditioners

Table 2. Mean, range (in parentheses) and coefficient of variation (CV%) of basic chemical properties total elemental composition of
mulches, soil conditioners (SC) and blended soil conditioners (blended SC) produced from composted garden organics in the
metropolitan areas of New South Wales
Within rows, mean values followed by different letters are statistically different (P < 0.05)

Parameter Mulch SC Blended SC Mulch SC Blended SC

(mean and range) (CV%)
Chemical properties
pH (1:5 solid :water) 6.53a (5.0–7.3) 6.89a (5.2–7.5) 7.13a (5.9–7.9) 11.1 11.3 7.7
Electric conductivity (dS/m) 1.22a (0.39–1.95) 1.96a (1.24–3.12) 3.08b (1.34–4.68) 40.7 20.5 50.9
Soluble P (mg/kg) 2.7a (0.9–5.3) 2.2a (0.9–7.1) 11.2a (0.2–43.4) 50.2 76.9 119.0
NH4-N (mg/kg) 13.2a (0.7–63.7) 6.6a (0.4–57.4) 78.0b (0.6–231) 164.4 140.8 107.7
NO3-N (mg/kg) 0.9a (0–2) 2.9a (0–16) 74.0b (0–527) 95.4 198.4 205.0
Mineral N (mg/kg) 26.7a (0.7–65.7) 15.8a (0.6–57.4) 202.1b (1.3–530) 159.5 98.4 106.5
C:N 71.7a (41.1–104.4) 25.5b (13.6–32.0) 19.9b (9.4–28.0) 27.0 26.6 37.5
Elemental composition
C (%) 44.6a (41–47) 25.1b (15–32) 20.6b (14–31) 4.6 25.8 29.3
N (%) 0.66a (0.5–1.0) 1.00b (0.9–1.2) 1.21b (0.5–2.1) 26.0 15.2 44.1
Ca (%) 0.71a (0.51–1.10) 1.44b (0.74–2.20) 2.23b (1.00–4.30) 28.2 33.3 54.2
K (%) 0.40a (0.14–0.52) 0.50a (0.38–0.57) 0.44a (0.12–0.75) 30.0 16.0 54.6
Mg (%) 0.14a (0.11–0.19) 0.25b (0.19–0.38) 0.29b (0.21–0.44) 21.4 24.0 24.1
Na (%) 0.14a (0.11–0.18) 0.14a (0.10–0.20) 0.16a (0.07–0.27) 21.4 21.4 31.3
P (%) 0.07a (0.03–0.13) 0.16b (0.05–0.24) 0.38b (0.09–0.82) 45.3 28.6 70.1
S (%) 0.11a (0.06–0.17) 0.13a (0.09–0.16) 0.19b (0.07–0.28) 36.4 15.4 31.6
Fe (%) 0.41a (0.19–0.76) 1.19b (0.70–1.50) 1.43b (0.90–2.60) 43.9 20.2 37.1
B (mg/kg) 11.9a (6.1–15.0) 11.9a (8.9–16.0) 10.2a (2.8–17.0) 33.1 23.0 47.2
Zn (mg/kg) 64.6a (48–87) 153.3b (120–200) 190.3c (63–330) 21.9 22.1 41.2
Cu (mg/kg) 39.4a (11–110) 59.6b (30–190) 84.8c (25–240) 89.2 84.4 75.5
1380 Australian Journal of Experimental Agriculture K. Y. Chan et al.

Table 3. FDA and phytotoxicity tests of soil conditioners (SC) and blended soil conditioners
(blended SC) produced from composted garden organics in the metropolitan areas of New South Wales
Mean values in the same row followed by different letters are statistically different (P < 0.05)

Parameter SC Blended SC SC Blended SC

(mean and range) (CV%)

FDA (µg fluorescein/g soil.min) 25.6a (10.7–40.1) 22.7a (8.9–52.9) 44.1 58.6
Root length (mm)A 98a (78–115) 63a (16–112) 13.7 65.8
AThe phytotoxity test using radish plant roots following Standards Australia (2003).

evaluated. The only exception was the blended soil conditioner
containing paper pulp (no. 14), which did not affect radish fresh
weight at any rate of application compared with the nil treatment
(Fig. 1a). It had the lowest total N concentration (0.5%) and
negligible mineral N (1.3 mg/kg; Table 2). Three of the blended
soil conditioners (no. 1, 11 and 17) showed continuing increases
in radish fresh weight with increasing application rates. Two of
the samples (no. 5 and 16) showed increases up to 50 t/ha and
then remained unchanged at higher application rates. Both had
high mineral N concentration but were found to be phytotoxic
(root length <20 mm long).
35
(a)
Fresh top weight (g/pot)
The nature of radish dry weight production in response to
increasing application rates of unblended and blended soil
conditioners was similar to that observed for fresh weight
production (Fig. 1). Regression of dry weight with mineral N,
total N and C/N ratio of the soil conditioners all resulted in
statistically significant correlation coefficients. Dry weight of
radish was found to be positively correlated with mineral N and
total N but negatively correlated with C/N ratio. The highest
correlation coefficient (r2 = 0.788) was obtained in the case of
C/N ratio. The equation is:
DW = 4.931 – (0.135 × CN) (r2 = 0.788, n = 10, P < 0.001) (1)
where DW is dry weight of radish at harvest and CN is C/N ratio
of the soil conditioners. Therefore, dry matter production of
11
radish was negatively related to C/N ratio of the soil
16 conditioners.
1

30
5
Discussion
25
17 The mulches had low nutrient concentrations (e.g. mean mineral
20 n = 27 mg/kg) and high C/N ratios (mean 72) suggesting they
are unlikely to be a source of nutrients for growing plants.
15 However, the mulches are likely to be suitable as compost
blankets for controlling weeds, conserving water and protecting
10 2 the soil surface against erosion (DEC 2005).
6
14 Compared with the other organic amendments (cow manure,
5 3
28
N = 1.5%, P = 0.5%, potassium (K) = 1.2%; blood and bone,
N = 5.3%, P = 5.2%; poultry manure, N = 3.1%, P = 2.5%,
0
K = 1.6%; sewage sludge, N = 2.0%, P = 1.0% (Burgess 1993)),
16
3.5 (b) soil conditioners produced from garden organics as the only
11
feedstock were generally low in total N (1%), P (0.07%) and K
3.0 1
Dry top weight (g/pot)

17
(0.5%). In fact, average total N of the soil conditioners was less
2.5
5 than a third of that of the poultry manure (Table 2). According
to Verdonck (1998), the total N of whole green compost (before
2.0 screening) ranged from 0.6 to 0.8%. Cook et al. (1998) used a
green waste compost with a total N concentration of 1% to grow
1.5 2 barley and found that the only significant yield increase was
6 observed at the highest rate of compost application (150 t/ha).
1.0 14 The low nutrient concentrations in the soil conditioners of our
3
study can be partly attributed to the nature of the feedstock,
0.5 28 which consisted of grass clippings and tree prunings. The mean
C/N ratio for the unblended soil conditioner was 25.5, which is
0 20 40 60 80 100 higher than the recommended optimal C/N range of 15 to 20 :1
for mature compost (Verdonck 1998). Composts with C/N ratios
Application rate (t/ha)
above 25 to 30 :1 usually immobilises inorganic N (Sullivan and
Fig. 1. (a) Fresh weight and (b) dry weight of radish tops (g/pot) in Miller 2001). The pot trial results confirmed the low nutrient
response to increasing application rates (0, 25, 50 and 100 dry t/ha) of soil availability of the soil conditioners produced from garden
conditioners () and blended soil conditioners () produced from organics alone and their unsuitability as soil amendments when
composted garden organics. applied as sole source of fertilisation. The significant reverse
Agronomic value of composted garden organics
relationship between dry matter of radish plants and C/N ratio
(Eqn 1) suggested the usefulness of the ratio as an indicator of
compost maturity. Garcia et al. (1992) also suggested that C/N
is a good indicator of maturity for composts made from
feedstock of high C/N ratio.
The FDA test is a measure of microbiological activity based
on enzymatic activities (Schnurer and Rosswall 1982).
Recently, the test has been evaluated as an indicator of
microbiological activity and hence of compost stability during
composting process, with conflicting results (Garcia-Gomez
et al. 2003; Saviozzi et al. 2004; Ntougias et al. 2006). While
Garcia-Gomez et al. (2003) concluded that the FDA test could
be used in place of respiration rate as a measure of stability
during composting of olive wastes, results of Saviozzi et al.
(2004) for composting of urban wastes indicated that FDA
values did not alter during the entire composting process and as
such was not a sensitive indicator of stability compared with the
standard respiration test. In the present study, similar FDA
results between the two categories of soil conditioners
suggested that they had similar level of microbiological activity,
even though 42% of the blended soil conditioners were found to
be phytotoxic as measured by the root length bioassay test.
Linear regression analyses indicated that the FDA results were
not significantly (P > 0.05) correlated with the root length
bioassay results and, therefore, suggested that it is unsuitable as
an indicator of compost stability. Further research is needed to
resolve the discrepancy and to assess the usefulness of FDA test
as an indicator of compost stability.
The high variability in the chemical composition of the CGO
compost products observed in this survey of NSW supported
earlier reports from other parts of Australia (Wilkinson et al.
2000; Wilkinson et al. 2002). Such high variability and,
therefore, uncertainty about the consistency of compost
products has been identified as a barrier to market acceptance in
Australian agriculture (Wilkinson et al. 2002). The variability in
the chemical composition of the CGO compost products is
probably related to the differences in the composition of the
organic feedstock and the degree of maturity of the products.
Garden organics can vary in total N concentrations depending
on the proportions of leafy material, such as grass clippings and
more woody material like tree trunks and branches. This is
influenced by several factors, such as the size of the operation,
geographical location, season, rainfall patterns and method of
garden organic collection (Wilkinson et al. 2002; Dorahy et al.
2005). For example, more grass clippings are generated during
the summer months, which can lower the C/N ratio in composts
produced during this period. Degree of maturity is determined
by the conditions and duration of the composting process (Chen
and Inbar 1993). Immature compost products are often
biologically unstable and have the potential to reduce crop yield
(Dick and McCoy 1993; Paulin et al. 2001). In our survey,
nearly half of the blended soil conditioners were found to be
phytotoxic and, therefore, immature. Results of the pot trial
highlighted the variable short-term plant responses, which were
a direct reflection of the variability of the soil conditioners.
CGO soil conditioners have the potential to play an important
role as a source of C for improving soil quality in the longer term
through increasing aggregate stability, water holding capacity
and microbial activity (Dick and McCoy 1993; Gibson et al.
Australian Journal of Experimental Agriculture 1381
2002). However, the unblended and blended CGO soil
conditioners evaluated in our survey had mean C concentrations
of 25 and 21%, respectively, and some were as low as 14–15%,
which was much lower than expected. This suggests inorganic
materials, such as sand and clay were somehow enriched during
processing. If these products are to be promoted for use as a
source of organic matter, then the total C concentrations will
need to be increased. The long-term benefit of CGO soil
conditioners in improving soil quality of organic matter
impoverished soil needs to be demonstrated in field experiments.
The generally higher variability of the blended soil
conditioners compared with the unblended soil conditioners
reflected the variety of materials used for blending. The survey
results further highlighted how the type of blending material
can influence the properties, such as nutrient status, and
agronomic performance of the final product. For example, the
blended soil conditioners containing biosolids were higher in N
and heavy metals than those containing other organic residuals.
The pot trial results emphasised that additional effort is still
required to improve product consistency, given that the soil
conditioners containing the same blending material
(e.g. poultry litter in no. 16 and 17) did not perform equally,
with respect to increasing radish growth (Fig. 1). Compost
quality and consistency can be improved by selecting feedstock
and blending materials more judiciously, as well as controlling
the composting process more carefully.
Farmers will only buy CGO soil conditioners and mulches if
there is a demonstrated value from using them, either through
improving soil quality and crop productivity or providing a net
economic return. Such information is not available and limits
the potential to develop agricultural markets for CGO.
Consequently, further research is required to quantify the
beneficial characteristics of these products and promote these
benefits to farmers and the broader community.
Conclusions
Our survey of CGO commercially produced in the metropolitan
areas of NSW has revealed that mulches and soil conditioners
have variable physical, chemical and biological properties.
Similarly, our pot experiment revealed the short-term
agronomic performance of a selection of unblended and
blended soil conditioners was inconsistent.
Product variability can be reduced and product quality can be
improved by more careful selection of feedstocks and blending
materials, as well as better control of the composting process.
Further research is required to quantify the benefits these
products have in terms of improving soil quality, crop
productivity and net economic returns to farmers. These issues
need to be addressed if viable agricultural markets for CGO are
to be developed.
Acknowledgements
Financial support provided by NSW Department of Environment and
Conservation is acknowledged. We also thank the compost producers for
their cooperation in undertaking this study.
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J. Plant Nutr. Soil Sci. 2008, 171, 893–899 DOI: 10.1002/jpln.200625199 893

Nitrogen retention and plant uptake on a highly weathered central

Amazonian Ferralsol amended with compost and charcoal
Christoph Steiner1,2*, Bruno Glaser1, Wenceslau Geraldes Teixeira3, Johannes Lehmann4, Winfried E.H. Blum5, and
Wolfgang Zech1
1 Institute of Soil Science and Soil Geography, University of Bayreuth, 95440 Bayreuth, Germany
2 Current address: Biorefining and Carbon Cycling Program, The University of Georgia, Athens, USA
3 Embrapa Amazonia Ocidental, CP 319–69011–970 Manaus, Brazil
4 Department of Crop and Soil Sciences, Cornell University, Ithaca, NY 14853, USA
5 Institute of Soil Research, University of Natural Resources and Applied Life Sciences (BOKU), 1180 Vienna, Austria

Abstract
Leaching losses of N are a major limitation of crop production on permeable soils and under
heavy rainfalls as in the humid tropics. We established a field trial in the central Amazon (near
Manaus, Brazil) in order to study the influence of charcoal and compost on the retention of N.
Fifteen months after organic-matter admixing (0–0.1 m soil depth), we added 15N-labeled
(NH4)2SO4 (27.5 kg N ha–1 at 10 atom% excess). The tracer was measured in top soil (0–0.1 m)
and plant samples taken at two successive sorghum (Sorghum bicolor L. Moench) harvests.
The N recovery in biomass was significantly higher when the soil contained compost (14.7% of
applied N) in comparison to only mineral-fertilized plots (5.7%) due to significantly higher crop
production during the first growth period. After the second harvest, the retention in soil was sig-
nificantly higher in the charcoal-amended plots (15.6%) in comparison to only mineral-fertilized
plots (9.7%) due to higher retention in soil. The total N recovery in soil, crop residues, and grains
was significantly (p < 0.05) higher on compost (16.5%), charcoal (18.1%), and charcoal-plus-
compost treatments (17.4%) in comparison to only mineral-fertilized plots (10.9%). Organic
amendments increased the retention of applied fertilizer N. One process in this retention was
found to be the recycling of N taken up by the crop. The relevance of immobilization, reduced N
leaching, and gaseous losses as well as other potential processes for increasing N retention
should be unraveled in future studies.

Key words: biochar Brazil / carbon / nitrogen cycling / slash-and-burn / soil organic matter / Terra Preta

Accepted January 2, 2008

1 Introduction tions such as charcoal. According to Duxbury et al. (1989)

The fertility of highly weathered Ferralsols in the tropics is
low, and soil organic matter (SOM) plays a major role in sus-
taining soil productivity. Thus, long-term intensive use is not
sustainable without nutrient inputs where SOM stocks are
depleted (Tiessen et al., 1994). Due to low nutrient-retention
capacity and high permeability of these soils, strong tropical
rainfalls cause rapid leaching of mobile nutrients such as
those applied with mineral N fertilizers (Hölscher et al.,
1997a; Giardina et al., 2000; Renck and Lehmann, 2004).
To overcome these limitations of poor soil, low nutrient-reten-
tion capacity and accelerated SOM decay require alternatives
to slash-and-burn (the prevalent agricultural practice in the
tropics) and alternative fertilization methods (Ross, 1993;
Fernandes et al., 1997). Instead of burning, involving a quick
release of nutrients (Kuhlbusch et al., 1991; Hölscher et al.,
1997b; Giardina et al., 2000; Hughes et al., 2000) and CO2
(Fearnside, 1997), fallow vegetation could be applied as
mulch (Sommer et al., 2004), compost, or charcoal (Lehmann
et al., 2002). Depending on the mineralization rate, organic
fertilizers such as compost, mulch, or manure applications
release nutrients in a gradual manner (Burger and Jackson,
2003). This may be different for very recalcitrant organic addi-
* Correspondence: Dr. Ch. Steiner;
e-mail: csteiner@engr.uga.edu
and Sombroek et al. (1993), it is important to separate effects
due to OM per se (maintenance and improvement of water
infiltration, water-holding capacity, structure stability, CEC,
healthy soil-biological activity) from those due to its decompo-
sition (source of nutrients).
Only relatively small amounts of charcoal are produced by
the traditional slash-and-burn technique. Charcoal represents
only 1.7% of the preburn biomass if a forest is converted into
cattle pasture (Fearnside et al., 2001). Producing charcoal for
soil amelioration from aboveground biomass instead of con-
verting it to CO2 through burning might be an alternative to
slash-and-burn (Lehmann et al., 2002; Steiner et al., 2004b;
Lehmann et al., 2006).
The existence of so-called “Terra Preta de Índio” (Indian black
earth) suggests that a human-induced accumulation of SOM
can be maintained over centuries (Sombroek et al., 1993).
These soils are exceptionally fertile, and their productivity is
most likely linked to an anthropogenic accumulation of P and
Ca associated with bone apatite (Lima et al., 2002) and black
C (BC) as charcoal (Glaser et al., 2001).

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894 Steiner, Glaser, Teixeira, Lehmann, Blum, Zech J. Plant Nutr. Soil Sci. 2008, 171, 893–899

The sustained fertility of charcoal-containing Terra Preta and
the frequent use of charcoal as a soil conditioner (Steiner
et al., 2004b) in Brazil provided the incentive to study the
effects of charcoal application on N cycling. Charcoal acted
as an adsorber which reduced N leaching in previous pot
experiments (Lehmann et al., 2002, 2003), and charcoal
additions proved to sustain fertility if an additional nutrient
source is given in a field trial (Steiner et al., 2007). Charcoal
plus fertilizer improved plant growth and doubled grain pro-
duction in comparison to the fertilizer without charcoal. Plant
biomass production sharply decreased within 1 y when only
mineral fertilizer was applied, but could be maintained for a
longer period of time when OM was added. The authors pro-
posed that charcoal can improve soil chemical, biological,
and physical properties, but could not completely discern the
mechanisms of fertility enhancement (Steiner et al., 2007).
Stable isotopes such as 15N can serve as valuable tracers to
study plant resource acquisition and as a means to understand
how plants interact with their abiotic and biotic environments
(Dawson et al., 2002). Therefore, our objective was to compare
the effect of organic amendments such as charcoal and com-
post on retention of 15N-labeled mineral fertilizer in a highly
weathered Ferralsol under humid tropical field conditions.
The charcoal derived from secondary forest wood and was
considered rather as soil conditioner than fertilizer due to the
charcoal’s low nutrient contents (Tab. 1). It was manually
crushed to particle sizes <2 mm. The applied 11 Mg ha–1
corresponded to the amount of charcoal-C which could be
produced by a single slash-and-char event of a tropical
secondary forest on Xanthic Ferralsols in central Amazonia
(Lehmann et al., 2002). The amount of C added with charcoal
was chosen as a reference value for adding the compost.
Compost was prepared from biomass of a secondary forest,
fruit residues, manure, and kitchen waste. On February 3,
2001, organic materials were mixed with hand hoes into the
top soil (0–0.1 m) of the plots, and the first mineral fertilization
was done on March 19, 2001 [30 N kg ha–1 as (NH4)2SO4,
35 kg P ha–1 as simple superphosphate, 50 kg K ha–1 as KCl,
and 2100 kg lime ha–1]. The fields were cropped twice (rice
Oryza sativa L. and sorghum Sorghum bicolor L. Moench)
prior the second fertilization on April 16 (see Tab. 1 for treat-
ment description). All crop residues remained on the field,
and only grains were removed. Sorghum was planted again
on April 18, 2002 in a density of 25 plants m–2 producing two
harvests by ratooning (July 21 and October 16, 2002). Only
these two harvests are subject of the present paper and
designated as 1st (HI) and 2nd (HII) harvest.

Table 1: Treatments, organic amendments, and harvest remnants

(from previous harvests remained in the field). Mineral fertilization (F)
2 Materials and methods
was applied after the second harvest in April 2002, 15N-labeled N
(55), P (40), K (50), lime (430) [kg ha–1].
2.1 Study location and experimental setup
Token Organic matter 1st 2nd Total
The experiment was conducted within a larger field trial estab- harvest harvest
lished 30 km N of Manaus, Amazonas, Brasil (3°8′ S, 59°52′ W, (HI) (HII)
40–50 m asl) at the Embrapa-Amazônia Ocidental (Empresa remnantsa remnantsa
Brasileira de Pesquisa Agropecuaria) experimental research [Mg dry matter ha–1]
station. The natural vegetation is evergreen tropical rainforest
COb compost (67) 5.13 1.11 6.24
with a mean annual precipitation of 2530 mm (1971–1997) hav-
ing its seasonal maximum between December and May, a F 1.98 0.27 2.25
mean annual temperature of 25.8°C (1987–1997), and relative CO + F compost (67) 6.24 1.51 7.75
humidity of 85% (Correia and Lieberei, 1998). The soil was clas- CC + F charcoal (11) 2.55 0.72 3.27
sified as a highly weathered Xanthic Ferralsol (FAO, 1990), fine-
½CC + ½CO + F charcoal (5,5), 4.74 1.10 5.85
textured with high clay content. It is strongly aggregated and has
compost (33,5)
medium contents of organic C (24 g kg–1), low pH values of 4.7
(in H2O), low CEC of 1.6 cmolc kg –1, and low base saturation CC + ½CO + F charcoal (11), 4.28 1.10 5.38
(BS) of 11% (Steiner et al., 2007). compost (33,5)

F = mineral fertilizer; CO = compost; CC = charcoal

This experiment is part of a long-term field trial established in a previous harvests July 7, 2001 and February 6, 2002
January 2001 (Steiner et al., 2007). The main objective was b unfertilized control treatment, reference value for 15N natural abun-

to study different organic-amendment combinations based on dance;

equal amounts of C additions. Fifteen different treatments
were established in five repetitions after clearing of approx.
3600 m2 secondary forest and removing the aboveground
biomass. The treatments were applied on 4 m2 plots (2 m × 2 m)
forming an entire field area of 1600 m2 (45 m × 35 m) with a
minimum distance to the surrounding vegetation of 10 m. For
this study, five mineral-fertilized (F) treatments were chosen,
four of them with compost (CO + F) and/or charcoal (CC + F)
application (Tab. 1). The treatment receiving compost only
(CO) served to provide a reference value for N isotope com-
position in soil and plants. This treatment was used as a con-
trol for all treatments as d15N did not differ between the treat-
ments without tracer application [(NH4)2SO4 fertilized or not].
previous mineral fertilization: March 2001 [kg ha–1] N (30), P (35), K
(50), lime (2100);
compost contained 10.1, 0.73, 2.85, 3.27, and 1.51 g kg–1 N, P, K,
Ca, and Mg, respectively;
charcoal contained 5.39, 0.03, 0.23, 0.82, and 0.17 g kg–1 N, P, K,
Ca, and Mg, respectively.
2.2 Tracer application, sampling, and calculations
The chosen treatments (F, CC + F, CO + F, ½CC + ½CO + F,
and CC + ½CO + F) received 15N isotope enrichment using
15N-labeled (NH ) SO with 10 atom% 15N excess. The tra-
4 2 4
cer was mixed in a ratio 1:1 with conventional (NH4)2SO4 and

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J. Plant Nutr. Soil Sci. 2008, 171, 893–899
applied at a rate of 55 kg N ha–1 in April 2002 (second fertili-
zation). Additionally, 40 kg P ha–1, 50 kg K ha–1, and 430 kg
ha–1 of lime was applied. Soil and plant samples were taken
at each harvest and analyzed for d15N. Only the top 0.1 m of
soil was sampled, this was also the depth down to which the
organic amendments were mixed. Two soil samples were
taken per plot to form one composite sample. Soils were air-
dried and ground before isotope analysis. Representative
plant samples were taken from the entire crop by using the
center 1.4 × 1.4 m (49 plants) of each plot. Plant samples
were dried at 70°C for 48 h and ground.
The remaining fertilizer N in soil or plant biomass was calcu-
lated after Eq. 1 (Boutton, 1996).
Nf
d15 Nf ˆ Nf
d15 Nc ‡Y
d15 NNPK , (1)
Nf = nitrogen content of biomass or soil in N-fertilized treat-
ment,
d15Nf = measured d15N value of biomass or soil in N-fertilized
treatment,
d15Nc = measured d15N value of biomass or soil in unfertilized
control treatment (only CO served as valid control treatment
as d15N did not differ between treatments without tracer appli-
cation),
d15NNPK = d15N of (NH4)2SO4 10 atom% 15N excess
(= 29330.3‰).
The amount of 15N remaining in soil or in plant biomass (Y)
was calculated according to Eq. 2. The subtraction of d15Nc in
the denominator was neglected because it is small (approx.
10 and 20 for soil and biomass, respectively) in comparison
to d15NNPK.
Nf
d15 Nf Nf
d15 Nc
Yˆ 15
. (2)
d NNPK
The percentage of N taken up by biomass or remaining in the
soil was calculated according to Eq. 3:
Y plots without organic amendment (9.7%). The compost treat-
N% ˆ
100, (3)
N…NH4 †2 SO4
where N(NH4)2SO4 = amount of tracer fertilized [27.5 kg ha–1
(NH4)2SO4 10 atom% 15N excess].
2.3 Analyses of soil and plant samples
Soil and plant samples were analyzed for their C and N con-
tents by dry combustion with an automatic C/N-Analyzer (Ele-
mentar, Hanau, Germay). Total N isotope composition in soil
and plants was determined using an Elemental Analyzer
(Carlo Erba NA 1500, Carlo Erba Reagenti, Rodano, Italy; for
Dumas combustion) connected to an isotope-mass spectro-
meter (FINNIGAN MAT delta E; Thermo Finnigan, San Jose
CA) via a split interface.
 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Nitrogen retention with compost and charcoal 895
2.4 Statistical analyses
Treatment effects were analyzed by one-way analysis of var-
iance (ANOVA). Significant treatment effects were detected
using the Fisher’s LSD (last significant difference) test. Statis-
tical analyses and figures were performed using SPSS 12.0
(SPSS Inc.) and SigmaPlot 7.0.
3 Results and discussion
While the soil C contents were significantly increased by the
organic amendments (CO and CC), the N values were only
significantly enhanced in comparison to the F plots if CO was
applied (Fig. 1). After the second harvest (HII), the N and C
contents on solely compost-amended plots (CO) did not differ
from those receiving only mineral fertilizer. The OM-amended
and N-fertilized plots had significantly higher C and N con-
tents than the F plots. All organic amendments significantly
increased the C : N ratio after the first harvest. This difference
was even more distinct after the second harvest but only on
charcoal-containing plots (Fig. 1).
Neither the N concentrations in the biomass nor the mea-
sured d15N values differed significantly (data not shown). The
significantly higher mineral N uptake by plants growing on the
compost-amended plots (CO + F) was due to a significantly
higher plant biomass production (Tab. 2).
Wardle et al. (1998) found greater tree-seedling growth, N
uptake, and enhanced efficiency of nutrient uptake in boreal
forest soils when charcoal was added. The authors assumed
that adsorption of phenolics by the charcoal diminished
adverse effects on plant growth, both as allelopathic agents,
and through complexing N, thus reducing its availability to
plants. In our case, soil analyses indicate that other nutrients
than N were more important to enhance plant growth leading
to N sequestration in biomass (Steiner et al., 2007). This
assumption is corroborated by the study of Alfaia et al. (2000)
who found only a 16% rice grain-yield increase due to
(NH4)2SO4 fertilization, but significant losses of fertilized N.
In the soil, the situation was rather different. After the second
harvest (HII), significantly more fertilizer N remained in the
soil amended with charcoal (15.6% of applied N) than on
ment showed intermediate values (12.6%) (Fig. 2). In the soil,
the increased retrieval of N rather than higher total soil N con-
tents caused the significantly enhanced N recovery. Only
remaining crop residues could have caused the increase in
encountered 15N from HI to HII. However, the treatment ½CC
+ ½CO + F showed a much larger increase in soil 15N
encountered from HI to HII than the F treatment but had
slightly less crop-residue input (Tab. 2). Only the compost
treatment (CO + F) generated significantly more crop residue
15N than the control group but this additional residue return
did not significantly affect the soil 15N at HII.
At least to some extent the accrued crop residues after HII
added to the soil’s N pool as shown by the shift in isotope
values (data not shown) because soil sampling was done
7–14 d after the harvest. Belowground biomass was not
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896 Steiner, Glaser, Teixeira, Lehmann, Blum, Zech J. Plant Nutr. Soil Sci. 2008, 171, 893–899

Figure 1: Contents of C and N in of the soil

at the first (HI) and second harvest (HII). The
error bars show the mean ± standard error.
The Fisher’s LSD (least significant differ-
ence) value is plotted to scale significant
mean separation (p < 0.05).

assessed and can contribute significantly to N cycling as the
root biomass pool can be assumed to be at least half as large
as the aboveground biomass pool (Lacerda et al., 2004;
Hattori et al., 2005). Lehmann and Zech (1998) found 30% of
sorghum roots in Kenya below 0.3 m. We sampled only the
upper 0.1 m, thus N could have been recycled from deeper
soil layers.
The significantly increased uptake of applied N by plants on
plots with additional compost and the significantly higher N
retention in the soil due to charcoal additions may be explained
by either reduced N leaching (Lehmann et al., 2003) or reduced
gaseous N losses (Yanai et al., 2007). Reduced N leaching may
be a result (1) of either improved retention of the applied NH‡
4 by
electrostatic adsorption to exchange sites provided by the com-

Table 2: Biomass production, N uptake, and withdrawal (grains) at the first (HI) and second harvest (HII). Different letters in the same column
indicate significant differences (p < 0.05) between treatments (Fisher’s LSD test, n = 5).

HI HII HI HII
crop residues grains crop residues grains N residues N grains N residues N grains
Treatment [Mg dry matter ha–1] [kg ha–1]
F 1.38 b 0.28 c 0.50 c 0.14 c 11.3 c 4.7 c 4.9 b 2.3 c
CO 1.50 b 0.28 c 0.56 c 0.16 bc 19.8 b 5.8 c 7.5 b 3.2 bc
CO + F 2.69 a 0.96 a 1.17 a 0.80 a 29.8 a 15.5 a 13.1 a 12.6 a
CC + F 1.85 b 0.42 bc 0.72 bc 0.28 bc 17.3 bc 7.1 c 7.0 b 4.6 bc
½CC + ½CO + F 1.76 b 0.65 b 0.80 abc 0.33 bc 18.9 bc 11.8 ab 8.7 b 6.1 bc
CC + ½CO + F 2.08 ab 0.49 bc 0.97 ab 0.49 ab 19.7 b 8.1 bc 9.1 ab 8.5 ab

F = mineral fertilizer; CO = compost; CC = charcoal

Crop residues remained in the field; mean C 44.8, N 9.97 g kg–1, C : N = 44.16; mean N content of grains = 16.9 g kg–1

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J. Plant Nutr. Soil Sci. 2008, 171, 893–899
post or charcoal or (2) of immobilization of N by microbial bio-
mass. Lehmann et al. (2003) made microbial immobilization
responsible for decreases in foliar N contents and total N uptake
as a response to charcoal additions due to their higher C : N
ratio. However, their pot experiment used larger amounts of
applied charcoal (67.6 and 135.2 Mg C ha–1) than our study. The
C : N ratio in the soil studied here was found to be significantly
higher in the charcoal treatments at both harvest times (Fig. 1)
in comparison to the control. Decomposition of a portion of
newly applied charcoal and concurrent N immobilization is most
likely a transient phenomenon as charcoal is much more stable
than other organic additions (Baldock and Smernik, 2002). In
the studied permeable soils under high-rainfall conditions,
temporary immobilization of mobile nutrients might be desirable.
The results of the 15N experiments by Burger and Jackson
(2003) suggest a very dynamic role of microbially bound N and
highlight the importance of N immobilization that is taking place
simultaneously with inorganic-N production by mineralization.
They concluded that greater C availability stimulates microbial
activity resulting in greater N demand, promoting immobilization
and recycling of NO3 . The resilience of soil C in charcoal-
amended plots shows the refractory nature of charcoal (Kuhl-
busch and Crutzen, 1995). While the stability of charcoal leads
to low C losses, nutrient release by mineralization is most likely
lower than from other organic materials. As charcoal is expected
to be an extremely recalcitrant form of OM, it is unknown to what
extent charcoal C favors N immobilization. In previous studies,
the same plots showed significantly increased plant growth and
potential for microbial-population growth in mineral-fertilized
soils amended with charcoal and in Terra Preta, but lower micro-
bial respiration in the absence of an easily degradable C source
(glucose) compared to soils without charcoal (Steiner et al.,
2004a, 2007). It has been shown that charcoal amendments
can significantly enhance nitrification in pine-forest soils
(DeLuca et al., 2006). The authors suggest that charcoal alters
 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Nitrogen retention with compost and charcoal 897
Figure 2: Amount of mineral N fertilizer
remaining in the soil (Ferralsol, a and b)
and in the crop (sorghum, c and d) after
the first (HI, a and c) and second harvest
(HII, b and d) as a percentage. Means
and standard errors (p < 0.05, Fisher’s
LSD test, n = 5).
the activity of the nitrifying community and removes inhibiting
phenols. We could not elucidate alterations in nitrification and its
possible consequences for leaching and denitrification,
although Birk (2005) found evidence that the organic amend-
ments increased the presence of anaerobic bacteria which pos-
sibly are denitrifiers. Dinkelmeyer et al. (2003) found a rapidly
decreasing total recovery (in plants and soil) of 15N (87% to 54%
to 24% at 1, 3, and 12 months after N application, respectively),
but they assessed a complex and deep-rooting agroforestry
system to a soil depth of 5 m. They found a minimum of 13% N-
fertilizer efficiency when applied to peach palm (Bactris gasi-
paes) and a maximum of 38% if applied to cupuassu (Theo-
broma grandiflorum). Alfaia et al. (2000) assessed N utilization
of a rice crop grown in central Amazona and found a N-fertilizer
efficiency of only 7.8%, 10.0%, and 1% in shoot, grain, and root
biomass, respectively. We found a maximum recovery in plant
biomass of 15% and total recovery of 22% 3 months after N
application at HI. Already after 4 d, Renck and Lehmann (2004)
found applied 15N in the soil solution to a depth of 0.6 m and to a
depth of 5 m after 1 week at the same study site. They found that
the largest part of applied 15N in the top soil was leached as
organic N. The total fluxes of organic N were similar to those of
NO3 in the topsoil. According to Lehmann et al. (2002), the NH‡ 4
adsorbability of charcoal is largely dependent on soluble OM, as
an addition of DOC from a manure extract increased NH‡ 4
adsorption. Therefore charcoal might have reduced leaching of
organic N in addition to the inorganic forms. The organic amend-
ments might likewise have altered denitrification but leaching is
most likely the predominant N loss in the studied soil and the
relative proportion could not be clarified in this study.
The charcoal’s low biodegradability (Kuhlbusch and Crutzen,
1995), low nutrient content (Ogawa, 1994; Antal and Grønli,
2003), and high porosity and specific surface area (Braida
et al., 2003) makes charcoal a rather exceptional SOM con-
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898 Steiner, Glaser, Teixeira, Lehmann, Blum, Zech
stituent. Terra Preta research has shown that oxidation on
the edges of the aromatic backbone and adsorption of other
OM to charcoal is responsible for the increased CEC, though
the relative importance of these two processes remains
unclear (Liang et al., 2006).
4 Conclusion
We conclude that both the higher retention of N in the soil
and the increased uptake by biomass are responsible for the
significantly enhanced N cycling in plots that received char-
coal. Soil charcoal amendments improve the efficiency of
mineral N fertilizer.
Acknowledgments
The research was conducted within SHIFT ENV 45, a Ger-
man–Brazilian cooperation and financed by BMBF, Germany
and CNPq, Brazil (BMBF No. 0339641 5A, CNPq 690003/
986). A financial contribution was given by the doctoral scho-
larship program of the Austrian Academy of Sciences. We
are grateful for the fieldworkers’ help particularly Luciana Fer-
reira da Silva and Franzisco Aragão Simão and the laborato-
ry technician Marcia Pereira de Almeida. Jago Birk helped
during an internship on the field and in the laboratory.
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J. Plant Nutr. Soil Sci. 2008, 171, 893–899 DOI: 10.1002/jpln.200625199 893

Nitrogen retention and plant uptake on a highly weathered central

Amazonian Ferralsol amended with compost and charcoal
Christoph Steiner1,2*, Bruno Glaser1, Wenceslau Geraldes Teixeira3, Johannes Lehmann4, Winfried E.H. Blum5, and
Wolfgang Zech1
1 Institute of Soil Science and Soil Geography, University of Bayreuth, 95440 Bayreuth, Germany
2 Current address: Biorefining and Carbon Cycling Program, The University of Georgia, Athens, USA
3 Embrapa Amazonia Ocidental, CP 319–69011–970 Manaus, Brazil
4 Department of Crop and Soil Sciences, Cornell University, Ithaca, NY 14853, USA
5 Institute of Soil Research, University of Natural Resources and Applied Life Sciences (BOKU), 1180 Vienna, Austria

Abstract
Leaching losses of N are a major limitation of crop production on permeable soils and under
heavy rainfalls as in the humid tropics. We established a field trial in the central Amazon (near
Manaus, Brazil) in order to study the influence of charcoal and compost on the retention of N.
Fifteen months after organic-matter admixing (0–0.1 m soil depth), we added 15N-labeled
(NH4)2SO4 (27.5 kg N ha–1 at 10 atom% excess). The tracer was measured in top soil (0–0.1 m)
and plant samples taken at two successive sorghum (Sorghum bicolor L. Moench) harvests.
The N recovery in biomass was significantly higher when the soil contained compost (14.7% of
applied N) in comparison to only mineral-fertilized plots (5.7%) due to significantly higher crop
production during the first growth period. After the second harvest, the retention in soil was sig-
nificantly higher in the charcoal-amended plots (15.6%) in comparison to only mineral-fertilized
plots (9.7%) due to higher retention in soil. The total N recovery in soil, crop residues, and grains
was significantly (p < 0.05) higher on compost (16.5%), charcoal (18.1%), and charcoal-plus-
compost treatments (17.4%) in comparison to only mineral-fertilized plots (10.9%). Organic
amendments increased the retention of applied fertilizer N. One process in this retention was
found to be the recycling of N taken up by the crop. The relevance of immobilization, reduced N
leaching, and gaseous losses as well as other potential processes for increasing N retention
should be unraveled in future studies.

Key words: biochar Brazil / carbon / nitrogen cycling / slash-and-burn / soil organic matter / Terra Preta

Accepted January 2, 2008

1 Introduction tions such as charcoal. According to Duxbury et al. (1989)

The fertility of highly weathered Ferralsols in the tropics is
low, and soil organic matter (SOM) plays a major role in sus-
taining soil productivity. Thus, long-term intensive use is not
sustainable without nutrient inputs where SOM stocks are
depleted (Tiessen et al., 1994). Due to low nutrient-retention
capacity and high permeability of these soils, strong tropical
rainfalls cause rapid leaching of mobile nutrients such as
those applied with mineral N fertilizers (Hölscher et al.,
1997a; Giardina et al., 2000; Renck and Lehmann, 2004).
To overcome these limitations of poor soil, low nutrient-reten-
tion capacity and accelerated SOM decay require alternatives
to slash-and-burn (the prevalent agricultural practice in the
tropics) and alternative fertilization methods (Ross, 1993;
Fernandes et al., 1997). Instead of burning, involving a quick
release of nutrients (Kuhlbusch et al., 1991; Hölscher et al.,
1997b; Giardina et al., 2000; Hughes et al., 2000) and CO2
(Fearnside, 1997), fallow vegetation could be applied as
mulch (Sommer et al., 2004), compost, or charcoal (Lehmann
et al., 2002). Depending on the mineralization rate, organic
fertilizers such as compost, mulch, or manure applications
release nutrients in a gradual manner (Burger and Jackson,
2003). This may be different for very recalcitrant organic addi-
* Correspondence: Dr. Ch. Steiner;
e-mail: csteiner@engr.uga.edu
and Sombroek et al. (1993), it is important to separate effects
due to OM per se (maintenance and improvement of water
infiltration, water-holding capacity, structure stability, CEC,
healthy soil-biological activity) from those due to its decompo-
sition (source of nutrients).
Only relatively small amounts of charcoal are produced by
the traditional slash-and-burn technique. Charcoal represents
only 1.7% of the preburn biomass if a forest is converted into
cattle pasture (Fearnside et al., 2001). Producing charcoal for
soil amelioration from aboveground biomass instead of con-
verting it to CO2 through burning might be an alternative to
slash-and-burn (Lehmann et al., 2002; Steiner et al., 2004b;
Lehmann et al., 2006).
The existence of so-called “Terra Preta de Índio” (Indian black
earth) suggests that a human-induced accumulation of SOM
can be maintained over centuries (Sombroek et al., 1993).
These soils are exceptionally fertile, and their productivity is
most likely linked to an anthropogenic accumulation of P and
Ca associated with bone apatite (Lima et al., 2002) and black
C (BC) as charcoal (Glaser et al., 2001).

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894 Steiner, Glaser, Teixeira, Lehmann, Blum, Zech J. Plant Nutr. Soil Sci. 2008, 171, 893–899

The sustained fertility of charcoal-containing Terra Preta and
the frequent use of charcoal as a soil conditioner (Steiner
et al., 2004b) in Brazil provided the incentive to study the
effects of charcoal application on N cycling. Charcoal acted
as an adsorber which reduced N leaching in previous pot
experiments (Lehmann et al., 2002, 2003), and charcoal
additions proved to sustain fertility if an additional nutrient
source is given in a field trial (Steiner et al., 2007). Charcoal
plus fertilizer improved plant growth and doubled grain pro-
duction in comparison to the fertilizer without charcoal. Plant
biomass production sharply decreased within 1 y when only
mineral fertilizer was applied, but could be maintained for a
longer period of time when OM was added. The authors pro-
posed that charcoal can improve soil chemical, biological,
and physical properties, but could not completely discern the
mechanisms of fertility enhancement (Steiner et al., 2007).
Stable isotopes such as 15N can serve as valuable tracers to
study plant resource acquisition and as a means to understand
how plants interact with their abiotic and biotic environments
(Dawson et al., 2002). Therefore, our objective was to compare
the effect of organic amendments such as charcoal and com-
post on retention of 15N-labeled mineral fertilizer in a highly
weathered Ferralsol under humid tropical field conditions.
The charcoal derived from secondary forest wood and was
considered rather as soil conditioner than fertilizer due to the
charcoal’s low nutrient contents (Tab. 1). It was manually
crushed to particle sizes <2 mm. The applied 11 Mg ha–1
corresponded to the amount of charcoal-C which could be
produced by a single slash-and-char event of a tropical
secondary forest on Xanthic Ferralsols in central Amazonia
(Lehmann et al., 2002). The amount of C added with charcoal
was chosen as a reference value for adding the compost.
Compost was prepared from biomass of a secondary forest,
fruit residues, manure, and kitchen waste. On February 3,
2001, organic materials were mixed with hand hoes into the
top soil (0–0.1 m) of the plots, and the first mineral fertilization
was done on March 19, 2001 [30 N kg ha–1 as (NH4)2SO4,
35 kg P ha–1 as simple superphosphate, 50 kg K ha–1 as KCl,
and 2100 kg lime ha–1]. The fields were cropped twice (rice
Oryza sativa L. and sorghum Sorghum bicolor L. Moench)
prior the second fertilization on April 16 (see Tab. 1 for treat-
ment description). All crop residues remained on the field,
and only grains were removed. Sorghum was planted again
on April 18, 2002 in a density of 25 plants m–2 producing two
harvests by ratooning (July 21 and October 16, 2002). Only
these two harvests are subject of the present paper and
designated as 1st (HI) and 2nd (HII) harvest.

Table 1: Treatments, organic amendments, and harvest remnants

(from previous harvests remained in the field). Mineral fertilization (F)
2 Materials and methods
was applied after the second harvest in April 2002, 15N-labeled N
(55), P (40), K (50), lime (430) [kg ha–1].
2.1 Study location and experimental setup
Token Organic matter 1st 2nd Total
The experiment was conducted within a larger field trial estab- harvest harvest
lished 30 km N of Manaus, Amazonas, Brasil (3°8′ S, 59°52′ W, (HI) (HII)
40–50 m asl) at the Embrapa-Amazônia Ocidental (Empresa remnantsa remnantsa
Brasileira de Pesquisa Agropecuaria) experimental research [Mg dry matter ha–1]
station. The natural vegetation is evergreen tropical rainforest
COb compost (67) 5.13 1.11 6.24
with a mean annual precipitation of 2530 mm (1971–1997) hav-
ing its seasonal maximum between December and May, a F 1.98 0.27 2.25
mean annual temperature of 25.8°C (1987–1997), and relative CO + F compost (67) 6.24 1.51 7.75
humidity of 85% (Correia and Lieberei, 1998). The soil was clas- CC + F charcoal (11) 2.55 0.72 3.27
sified as a highly weathered Xanthic Ferralsol (FAO, 1990), fine-
½CC + ½CO + F charcoal (5,5), 4.74 1.10 5.85
textured with high clay content. It is strongly aggregated and has
compost (33,5)
medium contents of organic C (24 g kg–1), low pH values of 4.7
(in H2O), low CEC of 1.6 cmolc kg –1, and low base saturation CC + ½CO + F charcoal (11), 4.28 1.10 5.38
(BS) of 11% (Steiner et al., 2007). compost (33,5)

F = mineral fertilizer; CO = compost; CC = charcoal

This experiment is part of a long-term field trial established in a previous harvests July 7, 2001 and February 6, 2002
January 2001 (Steiner et al., 2007). The main objective was b unfertilized control treatment, reference value for 15N natural abun-

to study different organic-amendment combinations based on dance;

equal amounts of C additions. Fifteen different treatments
were established in five repetitions after clearing of approx.
3600 m2 secondary forest and removing the aboveground
biomass. The treatments were applied on 4 m2 plots (2 m × 2 m)
forming an entire field area of 1600 m2 (45 m × 35 m) with a
minimum distance to the surrounding vegetation of 10 m. For
this study, five mineral-fertilized (F) treatments were chosen,
four of them with compost (CO + F) and/or charcoal (CC + F)
application (Tab. 1). The treatment receiving compost only
(CO) served to provide a reference value for N isotope com-
position in soil and plants. This treatment was used as a con-
trol for all treatments as d15N did not differ between the treat-
ments without tracer application [(NH4)2SO4 fertilized or not].
previous mineral fertilization: March 2001 [kg ha–1] N (30), P (35), K
(50), lime (2100);
compost contained 10.1, 0.73, 2.85, 3.27, and 1.51 g kg–1 N, P, K,
Ca, and Mg, respectively;
charcoal contained 5.39, 0.03, 0.23, 0.82, and 0.17 g kg–1 N, P, K,
Ca, and Mg, respectively.
2.2 Tracer application, sampling, and calculations
The chosen treatments (F, CC + F, CO + F, ½CC + ½CO + F,
and CC + ½CO + F) received 15N isotope enrichment using
15N-labeled (NH ) SO with 10 atom% 15N excess. The tra-
4 2 4
cer was mixed in a ratio 1:1 with conventional (NH4)2SO4 and

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J. Plant Nutr. Soil Sci. 2008, 171, 893–899
applied at a rate of 55 kg N ha–1 in April 2002 (second fertili-
zation). Additionally, 40 kg P ha–1, 50 kg K ha–1, and 430 kg
ha–1 of lime was applied. Soil and plant samples were taken
at each harvest and analyzed for d15N. Only the top 0.1 m of
soil was sampled, this was also the depth down to which the
organic amendments were mixed. Two soil samples were
taken per plot to form one composite sample. Soils were air-
dried and ground before isotope analysis. Representative
plant samples were taken from the entire crop by using the
center 1.4 × 1.4 m (49 plants) of each plot. Plant samples
were dried at 70°C for 48 h and ground.
The remaining fertilizer N in soil or plant biomass was calcu-
lated after Eq. 1 (Boutton, 1996).
Nf
d15 Nf ˆ Nf
d15 Nc ‡Y
d15 NNPK , (1)
Nf = nitrogen content of biomass or soil in N-fertilized treat-
ment,
d15Nf = measured d15N value of biomass or soil in N-fertilized
treatment,
d15Nc = measured d15N value of biomass or soil in unfertilized
control treatment (only CO served as valid control treatment
as d15N did not differ between treatments without tracer appli-
cation),
d15NNPK = d15N of (NH4)2SO4 10 atom% 15N excess
(= 29330.3‰).
The amount of 15N remaining in soil or in plant biomass (Y)
was calculated according to Eq. 2. The subtraction of d15Nc in
the denominator was neglected because it is small (approx.
10 and 20 for soil and biomass, respectively) in comparison
to d15NNPK.
Nf
d15 Nf Nf
d15 Nc
Yˆ 15
. (2)
d NNPK
The percentage of N taken up by biomass or remaining in the
soil was calculated according to Eq. 3:
Y plots without organic amendment (9.7%). The compost treat-
N% ˆ
100, (3)
N…NH4 †2 SO4
where N(NH4)2SO4 = amount of tracer fertilized [27.5 kg ha–1
(NH4)2SO4 10 atom% 15N excess].
2.3 Analyses of soil and plant samples
Soil and plant samples were analyzed for their C and N con-
tents by dry combustion with an automatic C/N-Analyzer (Ele-
mentar, Hanau, Germay). Total N isotope composition in soil
and plants was determined using an Elemental Analyzer
(Carlo Erba NA 1500, Carlo Erba Reagenti, Rodano, Italy; for
Dumas combustion) connected to an isotope-mass spectro-
meter (FINNIGAN MAT delta E; Thermo Finnigan, San Jose
CA) via a split interface.
 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Nitrogen retention with compost and charcoal 895
2.4 Statistical analyses
Treatment effects were analyzed by one-way analysis of var-
iance (ANOVA). Significant treatment effects were detected
using the Fisher’s LSD (last significant difference) test. Statis-
tical analyses and figures were performed using SPSS 12.0
(SPSS Inc.) and SigmaPlot 7.0.
3 Results and discussion
While the soil C contents were significantly increased by the
organic amendments (CO and CC), the N values were only
significantly enhanced in comparison to the F plots if CO was
applied (Fig. 1). After the second harvest (HII), the N and C
contents on solely compost-amended plots (CO) did not differ
from those receiving only mineral fertilizer. The OM-amended
and N-fertilized plots had significantly higher C and N con-
tents than the F plots. All organic amendments significantly
increased the C : N ratio after the first harvest. This difference
was even more distinct after the second harvest but only on
charcoal-containing plots (Fig. 1).
Neither the N concentrations in the biomass nor the mea-
sured d15N values differed significantly (data not shown). The
significantly higher mineral N uptake by plants growing on the
compost-amended plots (CO + F) was due to a significantly
higher plant biomass production (Tab. 2).
Wardle et al. (1998) found greater tree-seedling growth, N
uptake, and enhanced efficiency of nutrient uptake in boreal
forest soils when charcoal was added. The authors assumed
that adsorption of phenolics by the charcoal diminished
adverse effects on plant growth, both as allelopathic agents,
and through complexing N, thus reducing its availability to
plants. In our case, soil analyses indicate that other nutrients
than N were more important to enhance plant growth leading
to N sequestration in biomass (Steiner et al., 2007). This
assumption is corroborated by the study of Alfaia et al. (2000)
who found only a 16% rice grain-yield increase due to
(NH4)2SO4 fertilization, but significant losses of fertilized N.
In the soil, the situation was rather different. After the second
harvest (HII), significantly more fertilizer N remained in the
soil amended with charcoal (15.6% of applied N) than on
ment showed intermediate values (12.6%) (Fig. 2). In the soil,
the increased retrieval of N rather than higher total soil N con-
tents caused the significantly enhanced N recovery. Only
remaining crop residues could have caused the increase in
encountered 15N from HI to HII. However, the treatment ½CC
+ ½CO + F showed a much larger increase in soil 15N
encountered from HI to HII than the F treatment but had
slightly less crop-residue input (Tab. 2). Only the compost
treatment (CO + F) generated significantly more crop residue
15N than the control group but this additional residue return
did not significantly affect the soil 15N at HII.
At least to some extent the accrued crop residues after HII
added to the soil’s N pool as shown by the shift in isotope
values (data not shown) because soil sampling was done
7–14 d after the harvest. Belowground biomass was not
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896 Steiner, Glaser, Teixeira, Lehmann, Blum, Zech J. Plant Nutr. Soil Sci. 2008, 171, 893–899

Figure 1: Contents of C and N in of the soil

at the first (HI) and second harvest (HII). The
error bars show the mean ± standard error.
The Fisher’s LSD (least significant differ-
ence) value is plotted to scale significant
mean separation (p < 0.05).

assessed and can contribute significantly to N cycling as the
root biomass pool can be assumed to be at least half as large
as the aboveground biomass pool (Lacerda et al., 2004;
Hattori et al., 2005). Lehmann and Zech (1998) found 30% of
sorghum roots in Kenya below 0.3 m. We sampled only the
upper 0.1 m, thus N could have been recycled from deeper
soil layers.
The significantly increased uptake of applied N by plants on
plots with additional compost and the significantly higher N
retention in the soil due to charcoal additions may be explained
by either reduced N leaching (Lehmann et al., 2003) or reduced
gaseous N losses (Yanai et al., 2007). Reduced N leaching may
be a result (1) of either improved retention of the applied NH‡
4 by
electrostatic adsorption to exchange sites provided by the com-

Table 2: Biomass production, N uptake, and withdrawal (grains) at the first (HI) and second harvest (HII). Different letters in the same column
indicate significant differences (p < 0.05) between treatments (Fisher’s LSD test, n = 5).

HI HII HI HII
crop residues grains crop residues grains N residues N grains N residues N grains
Treatment [Mg dry matter ha–1] [kg ha–1]
F 1.38 b 0.28 c 0.50 c 0.14 c 11.3 c 4.7 c 4.9 b 2.3 c
CO 1.50 b 0.28 c 0.56 c 0.16 bc 19.8 b 5.8 c 7.5 b 3.2 bc
CO + F 2.69 a 0.96 a 1.17 a 0.80 a 29.8 a 15.5 a 13.1 a 12.6 a
CC + F 1.85 b 0.42 bc 0.72 bc 0.28 bc 17.3 bc 7.1 c 7.0 b 4.6 bc
½CC + ½CO + F 1.76 b 0.65 b 0.80 abc 0.33 bc 18.9 bc 11.8 ab 8.7 b 6.1 bc
CC + ½CO + F 2.08 ab 0.49 bc 0.97 ab 0.49 ab 19.7 b 8.1 bc 9.1 ab 8.5 ab

F = mineral fertilizer; CO = compost; CC = charcoal

Crop residues remained in the field; mean C 44.8, N 9.97 g kg–1, C : N = 44.16; mean N content of grains = 16.9 g kg–1

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J. Plant Nutr. Soil Sci. 2008, 171, 893–899
post or charcoal or (2) of immobilization of N by microbial bio-
mass. Lehmann et al. (2003) made microbial immobilization
responsible for decreases in foliar N contents and total N uptake
as a response to charcoal additions due to their higher C : N
ratio. However, their pot experiment used larger amounts of
applied charcoal (67.6 and 135.2 Mg C ha–1) than our study. The
C : N ratio in the soil studied here was found to be significantly
higher in the charcoal treatments at both harvest times (Fig. 1)
in comparison to the control. Decomposition of a portion of
newly applied charcoal and concurrent N immobilization is most
likely a transient phenomenon as charcoal is much more stable
than other organic additions (Baldock and Smernik, 2002). In
the studied permeable soils under high-rainfall conditions,
temporary immobilization of mobile nutrients might be desirable.
The results of the 15N experiments by Burger and Jackson
(2003) suggest a very dynamic role of microbially bound N and
highlight the importance of N immobilization that is taking place
simultaneously with inorganic-N production by mineralization.
They concluded that greater C availability stimulates microbial
activity resulting in greater N demand, promoting immobilization
and recycling of NO3 . The resilience of soil C in charcoal-
amended plots shows the refractory nature of charcoal (Kuhl-
busch and Crutzen, 1995). While the stability of charcoal leads
to low C losses, nutrient release by mineralization is most likely
lower than from other organic materials. As charcoal is expected
to be an extremely recalcitrant form of OM, it is unknown to what
extent charcoal C favors N immobilization. In previous studies,
the same plots showed significantly increased plant growth and
potential for microbial-population growth in mineral-fertilized
soils amended with charcoal and in Terra Preta, but lower micro-
bial respiration in the absence of an easily degradable C source
(glucose) compared to soils without charcoal (Steiner et al.,
2004a, 2007). It has been shown that charcoal amendments
can significantly enhance nitrification in pine-forest soils
(DeLuca et al., 2006). The authors suggest that charcoal alters
 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Nitrogen retention with compost and charcoal 897
Figure 2: Amount of mineral N fertilizer
remaining in the soil (Ferralsol, a and b)
and in the crop (sorghum, c and d) after
the first (HI, a and c) and second harvest
(HII, b and d) as a percentage. Means
and standard errors (p < 0.05, Fisher’s
LSD test, n = 5).
the activity of the nitrifying community and removes inhibiting
phenols. We could not elucidate alterations in nitrification and its
possible consequences for leaching and denitrification,
although Birk (2005) found evidence that the organic amend-
ments increased the presence of anaerobic bacteria which pos-
sibly are denitrifiers. Dinkelmeyer et al. (2003) found a rapidly
decreasing total recovery (in plants and soil) of 15N (87% to 54%
to 24% at 1, 3, and 12 months after N application, respectively),
but they assessed a complex and deep-rooting agroforestry
system to a soil depth of 5 m. They found a minimum of 13% N-
fertilizer efficiency when applied to peach palm (Bactris gasi-
paes) and a maximum of 38% if applied to cupuassu (Theo-
broma grandiflorum). Alfaia et al. (2000) assessed N utilization
of a rice crop grown in central Amazona and found a N-fertilizer
efficiency of only 7.8%, 10.0%, and 1% in shoot, grain, and root
biomass, respectively. We found a maximum recovery in plant
biomass of 15% and total recovery of 22% 3 months after N
application at HI. Already after 4 d, Renck and Lehmann (2004)
found applied 15N in the soil solution to a depth of 0.6 m and to a
depth of 5 m after 1 week at the same study site. They found that
the largest part of applied 15N in the top soil was leached as
organic N. The total fluxes of organic N were similar to those of
NO3 in the topsoil. According to Lehmann et al. (2002), the NH‡ 4
adsorbability of charcoal is largely dependent on soluble OM, as
an addition of DOC from a manure extract increased NH‡ 4
adsorption. Therefore charcoal might have reduced leaching of
organic N in addition to the inorganic forms. The organic amend-
ments might likewise have altered denitrification but leaching is
most likely the predominant N loss in the studied soil and the
relative proportion could not be clarified in this study.
The charcoal’s low biodegradability (Kuhlbusch and Crutzen,
1995), low nutrient content (Ogawa, 1994; Antal and Grønli,
2003), and high porosity and specific surface area (Braida
et al., 2003) makes charcoal a rather exceptional SOM con-
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898 Steiner, Glaser, Teixeira, Lehmann, Blum, Zech
stituent. Terra Preta research has shown that oxidation on
the edges of the aromatic backbone and adsorption of other
OM to charcoal is responsible for the increased CEC, though
the relative importance of these two processes remains
unclear (Liang et al., 2006).
4 Conclusion
We conclude that both the higher retention of N in the soil
and the increased uptake by biomass are responsible for the
significantly enhanced N cycling in plots that received char-
coal. Soil charcoal amendments improve the efficiency of
mineral N fertilizer.
Acknowledgments
The research was conducted within SHIFT ENV 45, a Ger-
man–Brazilian cooperation and financed by BMBF, Germany
and CNPq, Brazil (BMBF No. 0339641 5A, CNPq 690003/
986). A financial contribution was given by the doctoral scho-
larship program of the Austrian Academy of Sciences. We
are grateful for the fieldworkers’ help particularly Luciana Fer-
reira da Silva and Franzisco Aragão Simão and the laborato-
ry technician Marcia Pereira de Almeida. Jago Birk helped
during an internship on the field and in the laboratory.
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central Amazon? 17th World Congress of Soil Science, Bangkok,
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Introduction to the Pioneer Works of Charcoal Uses

in Agriculture, Forestry and Others in Japan

Makoto Ogawa

Osaka Institute of Technology

Introduction

In Asian countries, the highly intensive agriculture has been popular since ancient days
because of high population density, limited arable field and rice plant cultivation. Therefore,
various traditional cultivation techniques had developed in order to raise the productivity of
crops. Any kinds of wastes, the excretions of human and live stocks, straw, leaf litter,
grass、sewages and rice husk charcoal has been used as for the fertilizers and soil
amendments not only in agriculture but also gardening or greening. Particularly wood ash
containing some cinders was used as an important material for soil amendment and mineral
supply.

In Japan where the domestic supply of energy sources has been limited, the forest
resources, fire wood and charcoal were a most important energy source until the beginning of
20th century. Charcoal production and consumption increased depending on the increase of
population and reached to the maximum, 2.7 million ton per year in 1947. It may be estimated
that the wood, mainly broad leaved trees, of about 10 million ton was carbonized by the
traditional kiln at that time (1).

However, according to the rapid increase of the imported fossil fuels in 1960s, so called
“Fuel Revolution” progressed, and the productivity of charcoal decreased to the minimum,
about 30 thousand ton per year in 1980s. The abrupt changes in the people’s daily life and the
relations between human and forest caused the decrease of Matsutake mushroom,
mycorrhizal fungus and the outbreak of pine wilting disease (2).

In 1970s, under such a circumstance, late Dr. Kishimoto S. and Sugiura G. who were the
experts of charcoal and wood vinegar production began the movement to make revive charcoal
and to encourage the new use. In 1980 they published a book “ Introduction to charcoal
making on Sunday ” (3) and distributed the knowledge of popular charcoal making and use.
They contributed largely toward the present prosperity of charcoal business in Japan and
Asia. The author, Ogawa M. and his colleagues started the studies on the utilization of
charcoal in agriculture and forestry on their requests in 1980 and reported the effects of bark
charcoal on soy bean and pine tree in 1983 (4) (5).

Being encouraged by their activities and the extension of organic farming in Japan, the
application and studies of charcoal in agriculture began actively in 1980s. In 1986, “Technical
Research Association for Multiuse of Carbonized Materials” (TRA) was established by the
financial support of Forest Agency, and the studies of new charcoal use launched being
participated with 13 private companies for 3 years. The studies on the effects of charcoal and
wood vinegar were conducted covering the various fields; the improvement of carbonization

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technology, soil amendment in agriculture and greening, activation of microorganisms and

water purification. The research result with some general comments was published in 1990
and widely distributed (6).

By the report, the effects of charcoal and wood vinegar were recognized in public and
authorized as a specific material of soil amendment by MAFF (Ministry of Agriculture,
Forestry and Fishery) in1990. Contrary to the government expectation to raise the charcoal
production in rural area, the cheaper charcoals which are imported from Southeast Asia being
produced from coconut and oil palm shells has reached to the same amount with those in
domestic. In this paper, only the publications written by Japanese are introduced, because it
seems to be unavailable in other countries.

1. Charcoal Utilization in Agriculture and Gardening

1. Rice husk charcoal

The oldest description on charcoal use in agriculture is found in a text book, “ Nogyo Zensho
( Encyclopedia of Agriculture )” written by Yasusada Miyazaki in 1697 (7). He described in it;
“After roasting every wastes, the dense excretions should be mixed with it and stocked for a
while. This manure is efficient for the yields of any crops. It is called the fire manure”.
Probably the similar knowledge had been popular in China and Korea since ancient time.

In Asian countries, the rice husk charcoal which can be carbonized by simple method in field
soon after harvesting has been one of the most common materials for soil amendment. It
seems that the rice husk charcoal has been used for several thousand years since the
beginning of rice cultivation in Asia, because rice husk with high content of silica is hardly
decomposed and unavailable for the manure.

The custom to use rice husk charcoal mixed with the excretions had spread among farmers
particularly in wheat cultivation until about 60 years ago. It seems to be reasonable, because
the charcoal can absorb and keep chemical nutrients and deodorize the stench. However, this
method was too popular to investigate for scientists, and so the roles of charcoal in agriculture
had been neglected for long time. After the information on wood charcoal use circulated
in1980’s, the roles of rice husk charcoal have been noticed in the research field of agriculture.

Recently Ezawa T. et al. (8) reported that rice husk and its charcoal enhanced A (arbuscular)
mycorhiza formation of some crop plants and improved the soil physical properties when each
of them was added into top soil. Ishii T. et al. (9) also reported the same effects on the A
mycorrhiza formation of citrus seedlings. Komaki Y. et al. (10) suggested that a small amount
of rice husk charcoal was effective on the growth of Catharanthus roseus, but the browning of
leaves appeared with the excessive application because of high concentration of potassium
and higher pH than wood charcoal. Takagi K. et al. (11) proposed a practical method to reduce
the outflow of pesticides and herbicides from paddy field utilizing the absorbing ability of rice
husk charcoal. Also he tried to fix the bacteria with high decomposing ability of pesticide into
rice husk charcoal and succeeded to reduce the outflow from golf course (12).

 At present, the carbonizing method has developed from the traditional method to the
sophisticated facility equipped at the rice mill. Dried rice husk can be carbonized

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automatically and continuously in the self-burning kiln and the extra heat is utilized for small
scale power generation (13). Recently the facility and system have been exported to South
East Asia by the company. Probably it is possible to apply to the wheat and barley husks and
other crop plant residues.

In Asian countries, the studies of charcoal use in agriculture have been mostly carried out by
JICA experts. In Indonesia, Igarashi T. started the experiment to produce the rice husk
charcoal with simple method and to cultivate soy bean and maize with the charcoal in 1989
(14). Igarashi T. et al. (15) applied the charcoal together with magnesium phosphate and lime
respectively and tried to rotate soy bean with maize. The charcoal application enhanced root
nodule formation, plant growth and the yields significantly. The effect was sustained also in
the second crop of maize without any supply of fertilizers, and the after effect was observed up
to 10th rotation. In particular, the growth and yield of maize treated with the charcoal were
more than that in the control plot cultivated only by chemical fertilizer. He also tried to apply
the charcoal in several areas with different soil condition and reported that the effect seemed
to be different from the soil properties and the kind of crops (16). Since the publication of
experiment results, charcoal use is advancing in Indonesia.

In Thailand, Oka H. et al. (17) studied the effects of rice husk charcoal on the growth, yield
and nitrogen fixation rates of soy bean which was planted in the infertile sandy soil. He
reported that the amount of upper ground biomass, root, soy bean yield and the rate of
nitrogen fixation in soil increased significantly. In field, the application of 10 ton per hectare
was most efficient, and the effect appeared evidently in the second crop of sorghum and the
third of soy bean. The soil physical properties, porosity, water holding capacity, pH and CEC
were also improved, although the values were different depending on soil type.

In Philippine, Noguti A. et al. (18) confirmed the effect of rice husk charcoal on the growth of
beans in an infertile acidic red soil. In this experiment the rice husk charcoal of 2.5 t / ha and
lime of 1.5 t/ha were applied and mixed with top soil 15 cm in depth, chemical fertilizer was
applied and the root nodule bacteria inoculated. The number of root nodule and nitrogen
fixing rates increased in the plot with lime, but it was more stimulated evidently by the
addition of charcoal. The inoculation of root nodule bacteria and the application of rice husk
charcoal induced the same effect with single use of lime. It seems to be meaningful
economically in Philippine where the use of lime is rather costly for farmers.

2) Wood and bark charcoal

In 1983, Ogawa M. et al. (4, 5) reported that the bark charcoal powders containing a small
amount of chemical fertilizers were efficient for the mycorrhiza formation of pine tree and the
A mycorrhiza and root nodule formations of soy bean plant. In the experiment of soy bean
plant (19), the bark charcoal of broad leaved tree was mixed with 1 % (W / W) of the inorganic
fertilizer (N: P: K (8: 8: 8)), urea, super lime phosphate, ammonium sulfate and oil cake
powder respectively. These charcoal fertilizers were stocked for one week and scattered over
soil surface at 500 g / m2 and 1500g / m2 each before plowing. Soy bean seedlings without root
nodules were planted in each plot. The plots in which applied the inorganic chemical fertilizer,
100 g / m2 and 200 g / m2 and the control plot without any treatments were set to compare the
efficiency.

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Soy bean yields which were harvested from the plots with charcoal fertilizers of 500 g / m2
were mostly equal to those from the plots with 100 and 200 g of chemical fertilizer. By this
method the amount of chemicals could be saved remarkably to 1/20. Root nodule formation
was stimulated by charcoal fertilizers, but it was suppressed by the ones with ammonium
sulfate and synthetic chemical fertilizer. A mycorrhiza infection rates and the spore numbers
increased in the plots treated with charcoal fertilizers. It seems that the better growth of soy
bean plants resulted from the enhanced root growth and propagation of symbiotic
microorganisms being activated by the charcoal0..

The soil microbial flora in each plot changed with the application of charcoal fertilizers.
Charcoal fertilizers with higher pH than 8 inhibited the propagation of soil fungi, but
enhanced that of bacteria and Actinomycetes soon after the treatment and then returned to
the normal state gradually. It was prospected that the emission of carbon dioxide increased
temporarily being caused by the high microbial activity. At the same time, the free living
nitrogen fixing bacteria could be isolated on the nitrogen free medium. From the inoculation
test of the charcoal which was sterilized and buried in soil for a week, it was certain that the
charcoal became the habitat for root nodule bacteria (19).

The charcoal which was carbonized under high temperature is usually alkaline and porous
substance, and there is no substrate for saprophytic microorganisms. When the charcoal was
added into soil, at first the plant root grows toward the charcoal with enough water and air.
Next some microorganisms which can endure under high alkalinity can propagate in or
around charcoal. It seems that the charcoal provides a better habitat for the root and
symbiotic microorganisms.

These research results were confirmed also in the TRA. Wood charcoal was efficient to
improve the soil properties, but the mixtures with chemical fertilizers, zeolite, wood vinegar
and organic fertilizer exhibited the better effects than charcoal itself on tea plant, citrus and
vegetables (20), rice plant and apple tree (21) and some leguminous plants and grass for
greening (22).

It was found that root nodule bacteria could be immobilized with high frequency in the white
and hard charcoal with fine pores. So, Takagi S. (23) proposed the method to make the
inoculum of root nodule bacteria of leguminous plants utilizing the charcoal. On the other
hand, A mycorrhizal fungi showed the better growth on black charcoal which was carbonized
at 400-500 degree Celsius. The spore of Gigaspora margarita was formed in black volcanic soil
with high carbon content (24). The application of wood charcoal to the plant associated with
Frankia was effective also for the actinorhiza formation (25). In general, it is certain from
these results that the white charcoal with the fine pore and high pH is suitable for the
immobilization of bacteria and the black one for that of fungi. After the study of TRA Oohira
T. et al. (26) reported that the oak charcoal with finer pores than that of pine was suitable for
the immobilization of bacteria and Actinomycetes than that of pine. Meanwhile, the
immobilizing ability of pine charcoal could be improved by mixing of acetic acid. Matsubara Y.
et al. (27) reported that coconut shell charcoal and the inoculation of A mycorrhizal fungi were
effective to suppress the infection of soil born pathogen Fusarium spp..

The research results of charcoal use in various fields were distributed not only in Japan but
also in Asian countries mainly by Ogawa M. (28) (29) (30).

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3) Charcoal compost and wood vinegar

  In Japan, the compost making from litter and excretions and its use has been very
common for long time. Ash or carbonized material was an essential material to accelerate the
decomposition stimulating the bacterial activity and to neutralize the acidity. It is also well
known that the charcoal absorbs smell and liquid.

In 1980s, a private company invented the method to produce charcoal compost which was
made from fresh chicken dung and palｍ shell charcoal tip (31). In the process of compost
making, the more the charcoal is used, the faster the decomposition progresses with
exothermic reaction. Under aerobic condition Bacillus group became dominant and produces
antibiotics. Kobayasi (32) reported that these antibiotics inhibited the growth of some soil
born pathogens, Pythium, Rhizoctonia, Phytophtra and Fusarium and were effective to
suppress root diseases of various plants. At present the charcoal compost (Haipuro) has been
sold as a biological fungicide by the company. Following this instance, various kinds of organic
composts have been produced from other livestock excretions and charcoal and sold in market.

It had been also well known that the liquid flowing out of charcoal kiln was useful. Actually it
had been used in forestry nursery bed as a pesticide and at road side as a herbicide. Kishimoto
S. et al. (33) published a text book in which they recommended the utilization of wood vinegar
in agriculture, forestry, animal husbandry and food processing describing the methods for the
purification, distillation and filtration.

Wood vinegar is a byproduct which is obtained from the carbonization process cooling the
smoke by air or water. This liquid contains the volatile substances emitted with pyrolysis;
water soluble fraction is wood vinegar and the oily one is wood tar. The chemical composition
is different depending on raw materials. Major components of broad leaved trees are 81 %
water, acetic acid 8-10 %, methanol 2.44 %, acetone 0.56 % and soluble tar 7 %, and that of
conifers are rich in water, acetic acid 3.5 % and the others concentrations are lower than that
of broad leaved trees (34). The chemical components of wood vinegar containing many organic
substances are unstable, so it has been sold as the material complex.

It has been recognized since 1960s that the wood vinegars extracted from broad leaved trees
are more efficient for the growth and rooting of various plants than that of conifers. The
effects were confirmed also in the study of TRA (35) (36) (37). The concentrated liquid of wood
vinegar with strong acidity shows the effects to kill microorganisms, plants and some larvae,
but the diluted one to stimulate rooting, plant growth and microbial propagation. There are
many reports of the application in field practice and generally the effects have been well
known by users, but there are a few available scientific reports on the mechanisms because of
the chemical property.

2. Utilization in Forestry and Greening

1. Rehabilitation and reforestation of trees and pine forest by charcoal and

mycorrhiza

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In 1980, Ogawa et al. (38) tried to apply the bark charcoal powders with a small amount of
chemical fertilizers and succeeded to promote the growth of pine, Pinus thunbergii and
cultivate the mycorrhizal fungi, Rhizopogon rubescens which is a dominant mycorrhizal
species in the young stand with infertile basic sand

Some chemical fertilizers, urea, ammonium sulfate, super lime phosphate and synthetic
chemical fertilizer, were added to bark charcoal powder with 0.1-1.0% (W / W) respectively.
These materials were buried into the trenches 30 cm in depth and width after cutting the roots
and covered by sand. The regenerated fresh roots grew inside the charcoal layers vigorously
after 3 months and the mushrooms appeared abundantly along the trenches 9 months later.
After a year, the amounts of pine root and mycorrhiza increased remarkably in the charcoal
layers. In addition, the growth of shoots and the color of needles became better than before
treatment. These apparent effects probably resulted from the regeneration of root and the
formation of mycorrhiza. It might be caused from the enhanced nutrient uptake and the water
absorption through mycorrhiza. The water content in the charcoal was eminently higher and
it was kept 40 % even in mid summer comparing with 5 % outside the charcoal layer (39).

After releasing the experiment result the similar phenomena were confirmed by many
researchers in local forest experiment stations, because it is an edible mushroom in Japan.
For example Hirasa T. (40) devised the growing method of seedling with the mycorrhiza and
the cultivation method in field.

The same method has been widely accepted by professional gardeners and applied to various
kinds of tree species to make revive the famous trees in shrine, temple and park. Usually the
charcoal powder, maximum 1 cm in diameter, has been buried into trench or hole together
with a small amount of phosphate fertilizer and the spores of suitable mycorrhizal fungi to
host plant. Sometimes the root system was exposed removing top soil and covered by charcoal
powder as well. Also in nursery, the charcoal fertilizer is mixed with pot soil to improve soil
properties and inoculate the mycorrhizal fungi (41).

Both pine wilting disease by the insect and nematode and oak wilting by the insect and fungi
have been spreading mainly in western Japan for several decades, and now it has become so
serious problem in forestry. Pinus densiflora forest in low land mostly disappeared in the
southwest and reaches to northern most area of Honshu. Pinus thunbergii forest which had
been planted along the seacoast to prevent natural disasters was also declining. Therefore, the
practical methods of the rehabilitation and reforestation of pine forest have been expected
eagerly in rural areas.

Mycorrhiza formation is essential to Pinus species which generate the forests as a pioneer
plant at the dry site with infertile soil. These fungi also prefer to propagate in dry and infertile
soil condition. Therefore, it has been well known through the ecological study of pine forest (2)
that the man-made pine forest should be kept at the primary stage of plant succession by
cutting all of under shrubs and raking out the litter layer.

Ogawa M. proposed to rehabilitate and reforest the coastal pine forest using charcoal and
mycorrhizal fungi publishing a monograph (41). In the established forest, the same methods
described previously have been applied. Meanwhile, in the place from which all of pine tree
entirely destroyed, the under shrubs and top soil are completely removed before planting.

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Then pine seedlings with the inoculated mycorrhizal fungi, Rhizopogon, Pithoritus, Suillus
species, are planted together with the charcoal powder. The survival rates of these seedlings
were much higher than that without mycorrhiza and charcoal.

As such a trial, growing tree and burying charcoal, seems to be one of the practical methods
for carbon sequestration, Ogawa M. and his colleagues have promoted the movement to make
revive “White Sand and Green Pine” which is one of the symbolic scenery of Japan in order to
contribute the prevention of natural disasters and the countermeasure for global warming.

2) Rehabilitation of tropical rain forest and forestation in semiarid region

Ogawa M. who was working in the rehabilitation project of tropical rain forest in 1989 found
that a dominant species, Dipterocarpaceae, forms the ectomycorrhiza with several fungal
species among which Scleroderma columnare enhanced the seedling growth efficiently in
nursery. He tested the effect of charcoal powder on the growth of Shorea species and found
that the small amount of charcoal 2 % in volume was effective to stimulate the mycorrhizal
formation and the growth (42). Kikuti J. and M. Ogawa (43) devised the practical inoculation
method in which several saplings with the mycorrhiza were planted inside the nursery bed
and the pots were set under the canopy. By this method the mycelium of mycorrhizal fungi
penetrates through holes of the pot and naturally infected. Mori S. et al. (44) found the rice
husk charcoal is also effective and established the more convenient method. Rice husk
charcoal is not so harmful even if it was added too much.

The nursery technique to inoculate the mycorrhizal fungi with charcoal was also used in the
forestation project of pine in northern China and obtained the successful results (45). The
material such as charcoal which has higher water holding capacity is efficient to stimulate the
rooting and to supply water to the root through mycorrhiza even under severe dry condition. It
can be expected the charcoal will be used in dry land farming like date palm plantation and
the greening to stop the desertification (46).

3. Utilization of charcoal and wood vinegar in animal husbandry

and fishery

Charcoal powder had been commonly used to cure the intestinal disorders of animals. In
1980s the utilization of charcoal and wood vinegar extended into the fields of animal
husbandry and the fish aquaculture. One of the wood vinegar makers invented in 1970s the
tablet of charcoal powder containing wood vinegar and sold as a medicine of live stocks being
formally recognized MAFF (33). When animals take the drug with the feed, it is said that the
quality of meat, fat and egg can be improved being affected with the high activity of intestinal
microorganisms (47). Recently the use for pig and poultry increased to avoid the antibiotics
and to prevent the epidemics.

In general, charcoal powder has the strong ability to absorb the smell of excretions and
liquid. Especially the charcoal carbonized under lower temperature than 300 degree Celsius
shows the strong adsorption of ammonium (48). The mixture of charcoal and wood vinegar has
been used in barn as the deodorant and absorbent of liquid. It seems that these effects result
from the complex reactions of charcoal and wood vinegar, but there is a few available scientific

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data (33).

The material containing wood vinegar also is used in the aquaculture of eel and fish to keep
water clean (33). Sometimes high quality charcoal which is carbonized under higher
temperature has been also used for water purification in the fish tank. From experiences it is
said that fish likes to spawn around the charcoal, probably because some algae propagate on
the wood charcoal and carbon fiber

4. Trials of carbon sequestration by charcoal use in agriculture

and forestry
1. Developing charcoal industry

In 2000, the fundamental law concerning to the establishment of recycling social system was
enacted in Japan, and it was acquired to reuse any wastes as much as possible. Particularly
the incineration of waste woods was prohibited in order to reduce the discharge of CO2 and
dioxin. The total amount of waste woods mainly from construction has reached to 4.6 million
ton per year, but the domestic use is only 60 % still now. Therefore, some construction
companies have switched from the incineration to the carbonization and intended to use the
charcoal not only in agriculture but also for the humidity control of house and building,
because it is necessary to reduce the high humidity of traditional wooden building in Japan.

The function of charcoal for humidity control was studied intensively for several decades ago
(49) (50). Recently the construction companies have begun to spread the charcoal bag not only
over the under floor but above the ceiling (51). Meanwhile it was also reported that the
treatment was efficient to reduce an asthma and atopic dermatitis by diminishing the
population of molds and tick (52) (53).

According to the development of charcoal utilization, carbonization technology is developing

from the simple kiln to the automatic mass production facilities. The newly devised
carbonizers including various kinds of movable batch type kiln, rotary kiln, swing kiln and etc
has been sold for the mass production of waste wood charcoal. In some cases the extra gas has
been used for thermal electric power generation. At the same time, the studies to establish the
industrial standards and function of carbonized materials have started, and the charcoal
industry begins to be renewed in 2000s.

2. New materials of charcoal

Recently the raw materials of charcoal are ranging from waste woods to some flammable
substances. Among them the carbonization of bamboo and its utilization has been widely
noticed as one of the dealing methods of bamboo forest which is recently occupying the wide
ranges in southwest Japan. Bamboo charcoal with somewhat different structure and functions
from wood charcoal has been used for the purification of water and air.

Up to several years ago, all of garbage from urban life had been burned in the incineration
plant, but now some cities began to carbonize the garbage and try its utilization. But there are
some problems to be solved, because the water content is usually so high that it consumes too

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much energy for desiccation, and some products are unsuitable for agricultural use because of
the high concentration of heavy metals and salt (54). Therefore, the thermal electric power
plants have tried to burn the garbage charcoal mixing with coal and oil (55). The wastes
disposed from food processing and live stock excretions have been also carbonized and used in
agriculture with compost (55).

3) Carbon Sequestration by Forestation and Carbonization (CFSC)

On January 8 in 1991, Ogawa M. puts forward a new idea “The earth’s green saved with
charcoal” writing an article in Nihon Keizai Shinbun (Nikkei). He described in this paper the
outline of this concept; The carbon dioxide emitted into air can be fixed by photosynthesis into
the planted tree; If the waste wood will be carbonized and used in agriculture and other fields,
a vast amount of carbon will be stocked for long term in soil, and at the same time the
sustainable production of crops and trees will be realized.

In 1990s it has been recognized gradually in Japan that the carbonization are meaningful as a
counter measure against global warming through the production of renewable energy and the
uses for soil amendment in agriculture. However, the raw materials of carbonization in this
project must be restricted only the plant residues, because it induces large scale deforestation
as well as other biomass energy productions.

After COP3, Ogawa M. (56) (57) proposed the idea “Carbon Sequestration by Forestation
and Carbonization (CSFC Project)” in Japan and Asia. In Indonesia and Malaysia, the large
scale plantations of Acacia mangium for paper pulp and that of oil palm for biomass energy
are expanding after clear cutting of the natural forests. Fast growing species in monoculture
has been cut in rotation of every 8 to 10 years, and still now the slash burn fields are also
spreading in wide area. The sustainability of tree plantation and cropland in tropical region
has always become very serious problem. It needs to develop the more efficient techniques for
soil amendment to sustain the high productivity. It is certain that the charcoal use is one of
the most reliable and low-cost procedures being harmonized the nature. The same opinion was
proposed in the review written by Glaser B. et al. (58).

If charcoal is easily weathered in field, the idea may be meaningless. But it has been actually
exhibited that the charcoal has remained for one thousand years or more without weathering
at the ruin of ancient coin studio and in the old tombs in Japan and Korea.

After the observation of stability in sulfuric acid, sodium hydrate or under ultra violet light
for short term (personal communication), Kawamoto K. et al. (59) examined the durability of
wood charcoal against ozone and estimated the half-life of the charcoal in air. The weight of
saw dust charcoal carbonized at 400ºC was not affected with 8.5% ozone, while the one done at
1000ºC burned with 4.9% ozone. There were observed minute pores on the surface of charcoal
carbonized at 1000ºC. The half-life of charcoal which was carbonized at 1000ºC and treated by
ozone in air was about 50,000 years. From these results it is suggested that wood charcoal is
stable on a geological time scale.

On the other hand, Yamato M. et al. (60) reported the changes in soil chemical properties
and the crop yields when the bark charcoal of Acacia mangium, which was made of waste from
pulp industry, was applied as soil amendment for the cultivation of maize, cowpea and peanut

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in Sumatra, Indonesia. The yields of maize and peanut significantly increased after the
application of bark charcoal under the fertilized condition in an infertile soil. The amount of
root and the colonization rate of A mycorrhiza increased especially in maize. The application
of charcoal improved the soil chemical properties by neutralizing soil pH and increasing total
nitrogen, available phosphate contents, cation exchange capacity, amount of exchangeable
cations and base saturation. Moreover, it induced the decreasing of exchangeable Al ion which
seems to be harmful for the root growth. The effects of charcoal application appeared more
evidently in tropical region than those in Japan. Root nodule formations of leguminous plants
were also stimulated by charcoal.

Formerly it was reported that the population of free living nitrogen fixing bacteria
increased around the charcoal buried in tropical soil (28). Probably a small amount of nitrogen
seems to be accumulated into soil by charcoal application also in slash burn cultivation.

According to these results in Indonesia, the feasibility study of CSFC project was conducted
with the existing project of an industrial plantation and pulp production by Okimori Y. et al.
(61) in Indonesia Sumatra as an example of Clean Development Mechanism (CDM) project. It
was prospected that a total of 368,000 t / year of biomass residue and waste which were
disposed from plantation and pulp mill could be transformed into charcoal of 77,000 t / year, if
conventional charcoal producing methods are used. It was also expected that the carbon
emission reduction by the project reaches to 62,000 t-C / year (or 230,000 t-CO2). In addition,
this project can contribute to local economy providing the employment chance for 2,600
people, and will make realize the sustainable industrial plantation by soil amendment with
charcoal.

A similar trial was conducted also in Indonesian JICA project as a small scale example, and
the research result was reported by JICA (62). In this project it was confirmed that the
applications of charcoal were efficient both for the seedling growth of Acacia and the tree
growth in field.

In Western Australia, the multipurpose project of mallee eucalyptus plantation has been
carried out by Oil Mallee Company and Shea S. (63) in order to prevent the salinization of
arable soil. Another feasibility study of the project was conducted by Ogawa M. et al. (64) in
West Australia joining to the existing project. They proposed to carbonize wood waste and to
use the charcoal for soil amendment in wheat field. In this study, it was prospected that the
total carbon sink would reach 1,035,450 Mg-C with 14 % by aboveground biomass, 33.1 % by
belowground biomass and 52 % by charcoal in soil, if the plantation will be kept for 35 years.
Meanwhile, the effects of eucalyptus charcoal on the growth and yield of wheat were evidently
confirmed by Paul J. (in printing).

Ｔhe feasibility study of carbon sequestration in which various kinds of waste wood out of
construction, saw mill, trimmed branch and others were recycled by carbonization was
conducted by RITE (Research Institute of Environment and Industry) as summarized in the
previous paper(64). It was focused on the effective use of surplus heat from a garbage
incinerator for carbonizing woody materials. It was prospected from the study that the waste
wood of 936.0 Mg-C / year would be converted into the net carbon sink of 298.5 Mg-C / year by
carbonization, with the fixed carbon recovery of the system being 31.9 %.

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The life cycle assessment has been conducted also in Japan on the case of carbonization of
waste wood and the use of charcoal for construction and greening. By this study it was
certified that the CSFC project is significant and useful for carbon sequestration also in
carbon and energy balance (unpublished).

From these research results, it is expected that CSFC project will be recognized as a
concrete, easy and low cost counter measure when the global warming will intensify more
seriously in the near future. Today the charcoal uses in various fields are extending through
the release of technical informations, and the charcoal production industry is growing as one
of environment businesses not only in Japan but also among Asian countries.

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 J. Japan. Soc. Hort. Sci. 63(3) : 529-535. 1994.

Effects of Charcoal as a Soil Conditioner on Citrus Growth and

Vesicular-Arbuscular Mycorrhizal Development

Takaaki Ishii1 and Kazuomi Kadoya2

I Faculty of Education , Ehime University, Matsuyama, Ehime 790
2College of Agriculture
, Ehime University, Matsuyama, Ehime 790

Summary

Effects of several kinds of charcoal applied to soil on citrus growth and vesicular-
arbuscular mycorrhizal (VAM) development were investigated. Satsuma mandarin (Citrus
unshiu Marc.) trees on trifoliate orange (Poncirus trifoliata Raf.) rootstocks were trans-
planted to root boxes using the soil mixed with charcoal derived from rice husk, citrus
juice sediment or western spruce bark. The trees were inoculated with the spores of Glomus
fasciculatum (Thaxter) Gerdemann and Trappe emend. Walker and Koske. Elongation of the
roots in the charcoal treatments was more vigorous than that in the charcoal-free control.
The fresh weigths of the root, shoot and the whole tree increased in response to charcoal
application. The intensity of VAM infection in any charcoal treatment was higher than
that in the control. In particular, the percentage of the infection in the rice husk charcoal
plot was 41.5 and P concentration in the leaf exceeded that of the control.
In a Citrus iyo orchard, the percentage of VAM infection was 52% in the rice husk
charcoal plot, the highest among plots. The intensity in the Bahia grass (Paspalum
notatum Flugge.) plot was next, followed by the third highest rate found in the abandoned
plot which had not been cultivated in recent years. The lowest percentage of VAM infec-
tion was in a clean-culture plot. A microscopic observation also revealed that in a
charcoal-treated plot there were many sites where VAM fungi infected the root.

them (Dixon et al., 1988; Edriss et al., 1984; Ishii

Introduction
et al., 1992b; Menge et al., 1978; Nemec, 1979).
A classic book written in Japan (Miyazaki, These fungi improve mineral nutrition of the host
1697) explained that soybean plants vigorously by increasing P uptake from a P deficient soil
flourished with a minimum of care when their (Antunes and Cardoso, 1991; Ferguson and
seeds were sowed with charcoal. Recently, Ogawa Menge, 1986; Graham and Timmer, 1985; Krikun
(1987) reported that charcoal applied to the soil and Levy, 1980; Nemec, 1979; Tang et al., 1984).
could stimulate the activity of soil microorganisms Higher concentration of minor elements, especially
and promote the formation of root nodules and Zn (Krikun and Levy, 1980) and Cu (Timmer and
vesicular-arbuscular mycorrhizae in soybean Leyden, 1980), were also observed after an inocu-
roots. lation with VAM fungi. Because the fungi pro-
VAM symbioses are exceptionally common vided essential elements for citrus growth, the in-
among terrestrial flowering plants (Harley and fected trees could grow more rapidly and appeared
Harley, 1987). Among these plants, there is a wide healthier than non-infected trees. This phe-
range of dependency on VAM fungi for plant nomenon was especially noticeable in soils of low
growth. Citrus is also infected by several kinds of fertility (Nemec, 1979). Furthermore, VAM fungi
VAM fungi and is considered highly dependent on inoculation may increase tolerance to water stress
by regulating stomatal opening through hormone
Received for publication 7 February 1994.
Parts of this paper were presented at the 1989 Spring synthesis (Graham et al., 1987). In our reports,
Meeting and 1990 Spring Meeting of the Japanese Society the photosynthesis and transpiration rates of
for Horticultural Science. VAM fungi-infected satsuma mandarin trees grow-

529
 530 T. Ishii and K. Kadoya

ing in P-deficient soil surpassed those of non-VAM coal were mixed in a proportion of 50 to 1 by

trees stressed by high temperatures in August weight. The pH and electric conductivity (EC) of

(Shrestha et al., 1992). Interestingly, an inocula- the mixtures were measured with a pH meter and

tion of VAM fungi improved the fruit quality of an EC meter, respectively.

satsuma mandarin trees. In particular, it enhanced Two months after planting, the roots were

the Hunter's a/b value of peel color and the sugar treated with 50 g fresh weight of soil inoculum con-

content in juice (Ishii et al., 1992b). In the citrus taining 300 ~ 340 spores of Glomus fasciculatum .

orchards where high quality fruit was produced, The inoculum was obtained from greenhouse pot

the percentage of VAM infection in the root was cultures of Bahia grass inoculated with Glomus

very high (Shrestha et al., 1993). fasciculatum originally isolated from citrus

On the other hand, phytotoxic substances exist orchards in Matsuyama city, Ehime prefecture,

in the bark and sawdust extracts from several Japan. In 1988, each tree was fertilized with 6.4 g

woody forest species, especially in the bark ex- of N, 3.2 g of P, and 3.8 g of K per annum from a

tracts of hinoki cypress (Chamaecyparis obtusa mixture of ammonium sulfate, calcium phosphate,

Sieb. et Zucc. ex Endl.) which are condensed tan- and potassium sulfate, respectively; the P and K

nins (Ishii and Kadoya, 1993). In soils to which contents in the charcoal were first deducted. The

unfermented organic matter were added, ethylene control trees were supplied with 5 g of calcium

has often been detected at concentrations high carbonate-magnesium sulfate mixture (8 : 2 by

enough to inhibit the growth of citrus trees (Ishii weight) to improve the soil pH. A Hoagland minor

and Kadoya, 1984). The problem of growth inhibi- element solution (1 liter/tree) was administered to

tion is solved, however, if the organic matter with each trees. In 1989, P was excluded.

phytotoxic substances is first transformed into Roots appearing on the glass plates of root boxes

charcoal and then used as a soil amendment. were traced onto transparent plastic sheets from

The purpose of this study is to investigate which the root lengths were measured, using a

effects of several kinds of charcoal applied to soil personal computer equipped with an image proc-

on citrus growth and VAM development. essor (NEC mediagraph MG -10 with a stylus pen,

Tokyo, Japan) and a special software program.

Materials and Methods
In early December of 1989, the trees were re-

moved from the root boxes, and then the total,

Experiment 1. Effects of charcoal application on citrus
root, and shoot fresh weights were measured. For
growth and VAM development
the determination of leaf P, leaf samples were

In this experiment, we examined the effects of ashed at 550 •Ž overnight, and the residues were

charcoal application on the growth and VAM de- dissolved in 2.4 N HC]. The P content was meas-

velopment of 'Aoshima' satsuma mandarin trees on ured colorimetrically by the method of Deniges

trifoliate orange rootstocks for two years. In early (1920, 1921). Undamaged feeder rootlets were

April of 1988, three two-year-old satsuma man- sampled and rinsed with distilled water for a few

darin trees per plot were transplanted individually seconds. After the rootlets were cut into 2 - cm seg-

to root boxes (40 cm × 40 cm × 40 cm) containing ments behind the growing tip, the segments were

the mixtures of river sand and a specific charcoal. immediately fixed in FAA (formalin: acetic acid:

Before planting, the roots were carefully washed 50% ethanol, 13 : 5 : 200, v/v/v). Ten segments

to remove the soil which had nourished the trees. per treatment, stained by the technique of Phillips

The control soil lacked charcoal. The charcoal and Hayman (1970), were analyzed for the intensi-

used was made by using a chimney (15 cm in dia- ty of VAM infection by light microscopy. The per-

meter and 1.8 m in length) with some holes for centage of VAM infection was calculated with the

aeration or an oil drum (200 liter) equipped with following equation :

a chimney (15 cm in diameter and 1.8 m in

length). The charcoal sources were rice husk, cit-
rus juice sediment and western spruce bark. The
charcoal derived from western spruce bark was
broken into 5 ~ 10 mm pieces. The sand and char-
 J. Japan. Soc. Hort. Sci. 63(3) : 529-535. 1994. 531

13 times higher than that of the control plot

Experiment 2. Soil management and the intensity of
(Table 1). This is because NaCl permeated into the
VAM infection in citrus roots
bark during sea storage after being imported into
In late April of 1987, 4 experimental plots of 5 Japan from North America.
trees each were prepared in a Citrus iyo (15 - year- Soils treated with 3 kinds of charcoal had signif-
old trees on trifoliate orange rootstock) orchard in icant effects on growth, leaf P concentration, and
Matsuyama city, Ehime prefecture. The ex- VAM development in roots of satsuma mandarin
perimental plots were as follows : 1) charcoal as a trees. About 2 months after the onset of this ex-
soil amendment, 2) abandoned culture, 3) sod cul- periment, except in the western spruce bark char-
ture with Bahia grass, and 4) clean culture by us- coal treatment, roots appeared on the glass plates,
ing herbicides such as paraquat dichloride and and their elongation rates indicated that roots in
N- (phosphonomethyl) glycine (3 times in a year). the charcoal-treated plots were more vigorous than
The charcoal plot had two pits (60 × 60 cm in ones in the control. As of November 8, 1989, the
width and 40 cm in depth) circling a tree and filled root length in any charcoal treatment was about
with 6 kg of rice husk charcoal. Paraquat dichlo- 1.5 times longer than that in the control. The total
ride was applied once annually. In the Bahia grass fresh weights and the fresh weights of roots and
plot, the grass was mowed once each summer. Ex- shoots increased with charcoal treatments. The
cept for the abandoned plot, the rest received 32 growth increments varied little among the kinds of
kg N, 23 kg P, and 25 kg K per 10 a annually. The charcoal (Table 2). The intensity of VAM infection
application of agrochemicals, such as fungicides in any charcoal treatment was higher than that in
and pesticides, followed the guidelines of disease the control; that of the rice husk charcoal treat-
and pest control for Ehime prefecture. ment attaining 41.5% (Table 3). Hardly any signif-
In early September of 1988, root samples were icant differences in leaf P concentration among
obtained from 3 to 5 places of each plot at a depth treatments with western spruce bark charcoal, cit-
of 5-10 cm, and then the intensity of VAM infec- rus juice sediments charcoal and the control were
tion was determined by the methods described observed; but leaf P concentration in the rice husk
above. The root structure was observed with a charcoal treatment, which significantly stimulated
scanning electron microscope (SEM, JEOL type VAM infection, was higher than that in the control
JSM - T200, Tokyo, Japan). The apical 20 mm of (Table 3).
20 elongating roots from each plot were rinsed
Experiment 2.
with distilled water for a few seconds and the
apices were immediately fixed in Karnovsky solu- The intensiy of VAM infection in the rice husk
tion (Karnovsky, 1965) at room temperature for charcoal plot was 52%, the highest among plots.
24 hr. After being dehydrated through graded The intensity in the Bahia grass sod plot was
solutions of ethyl alcohol-acetone, they were second highest, whereas that of the abandoned plot
divided into 4 segments in 100% acetone. These was third. The lowest percentage of VAM infec-
segments were then immersed in acetone for 2 hr, tion was in the clean culture plot where herbicides
critical-point-dried, mounted on aluminum stubs were used 3 times a year (Table 4). The hyphae,
with silver conducting paint, and coated with a vesicles and arbuscles of VAM fungi were fre-
thin layer of gold using an ion-coater (Eiko
Engineering type IB -2, Tokyo, Japan). The roots
were observed in a SEM and photographed.
Table 1. The pH and electric conductivity (EC) of soils
Results treated with charcoal (Experiment 1) .

Experiment 1.

No differences in soil pH among treatments were

observed. The EC value of the charcoal treatments
was higher than that of the control. The EC in the
western spruce bark charcoal treatment was about
 532 T. Ishii and K. Kadoya

Table 2. Effect of charcoal application on the growth of satsuma mandarin trees (Experiment 1) .

Table 3. Effect of charcoal application on vesicular-

arbuscular mycorrhizal (VAM) development and
leaf phosphorus (P) concentration in satsuma man-
darin trees (Experiment 1) .

Table 4. Effect of soil management on VAM development

in Citrus iyo trees (Experiment 2) .

quently observed on/in citrus roots sampled from Fig. 1. Photomicrograph of VAM fungal structures in
the charcoal-treated plots (Fig. 1). The SEM photo-
Citrus iyo roots stained with typan blue.
micrographs also indicated that in the charcoal- a : fungal hyphae ( × 100), b : vesicle ( × 150), c :
treated plot there were many sites where VAM arbuscle ( × 600).
fungi infected and penetrated into the root (Figs. 2
and 3).
ment in the root were stimulated by applying char-
Discussion
coal to soil. This stimulation of citrus growth by
In Japan, it has long been known that charcoal charcoal is attributed to an increase in the percen-
is a very effective soil conditioner which promotes tage of VAM infection in the roots. Ogawa (1987)
plant growth. Charcoal application may result in also reported that the enhanced colonization by
improving physical properties of soil, its fertility, symbiotic microorganisms, such as Rhizobium and
and biological conditions. The present experiment VAM fungi, by charcoal application, invigorated
indicated that citrus growth and VAM develop- soybean plants.
 J. Japan. Soc. Hort. Sci. 63(3) : 529-535. 1994. 533

Fig. 2. SEM photomicrograph of VAM fungi in Citrus iyo roots.

Left : × 200, Right : × 1000.

gi by fungicides such as thiophanate methyl, be-

nomyl, iprodione, and copper fungicides is severe.
In the case of herbicides, Kobayashi (1988)
showed that the germination of Gigaspora margari-
ta spores was severely repressed by 48 ppm para-
quat dichloride or 410 ppm N- (phosphonomethyl)
glycine. In our experiment, the percentage of VAM
infcetion in the herbicide-treated clean culture plot
is lower than that of the abandoned plot.
The pH value of water extracts from charcoal
was high (Ishii and Kadoya, 1990), indicating that
charcoal ameliorated soil acidity. Generally, soil
pH is low in citrus orchards in Japan, so that the
percentage of VAM infection in the root is low and
the number of VAM spores in the soil is small
(Ishii et al., 1989b, 1992a). By neutralizing soil
acidity, charcoal may be improving the growth and
development of VAM fungi.
Fig. 3. SEM photomicrograph of a VAM spore There are very few reports on VAM develop-
(Glomus spp.) and invasion of its hyphae ment in citrus trees grown in Japan. When roots of
into roots (× 1000).
satsuma mandarin and Citrus iyo trees from 24
orchards in Ehime prefecture (in southwestern
Japan) were observed for VAM infections, they
The increased VAM infections by charcoal ap- were not extensive except for an orchard with
plication may be because charcoal absorbs many good soil conditions which produced 9 ~ 10 t satsu-
kinds of toxic substances and agrochemicals which ma mandarin fruit per 10 a every year (Ishii et al.,
inhibit root growth and microbial activity. It has 1989b). Numerous VAM spores in the soil and
also been shown that some agrochemicals inhibit VAM -infected plants are generally observed in
the germination of VAM spores (Kobayashi, 1988; woodlands and non-cultivated fields. This indi-
Ogawa, 1987). The growth inhibition of VAM fun- cates that there are many factors which restrict
 534 T. Ishii and K. Kadoya

the existence and growth of VAM fungi in Soil 131 : 11-19.

orchards because of our present soil management Deniges, G. 1920. Reaction de coloration extremement
sensible des phosphates et des arseniates. Compt.
practices, such as the usage of agrochemicals and
rend. 171 : 802-804.
chemical fertilizers. The average annual amount of
Deniges, G. 1921. Determination quantitative des plus
P applied is about 20 kg per 10 a. That P, espe-
faibles quantites de phosphates dans les produits
cially soluble P, is detrimental to VAM develop- biologiques par la methode ceruleomolybdique.
ment in citrus roots was reported earlier (Antunes Compt. rend. Soc. biol. 84 : 875-877.
and Cardoso, 1991; Graham and Timmer, 1985). Dixon, R. K., H. E. Garrett and G. S. Cox. 1988. Cyto-
Several kinds of VAM fungi, however, live in our kinin activities in Citrus jambhiri Lush. seedlings
soil in spite of many malpractices in our present colonized by vesicular-arbuscular mycorrhizal
soil management (Ishii et al., 1992a). We sug- fungi. Tree 2 : 39-44.
Edriss, M. H., R. M. Davis and D. W. Burger. 1984. In-
gested that VAM formation in citrus roots could
fluence of mycorrhizal fungi on cytokinin produc-
be effectively increased through application of
tion in sour orange. J. Amer. Soc. Hort. Sci. 109 :
charcoal to the soil or introduction of a sod cul-
587-590.
ture system. In particular, the application of char- Ferguson, J. J. and J. A. Menge. 1986. Response of cit-
coal is very effective for VAM development. Con- rus seedlings to various field inoculation methods
trarily, an excess of charcoal inhibits citrus with Glomus deserticola in fumigated nursery soils.
growth (Ishii and Kadoya, 1990). This inhibition J. Amer. Soc. Hort. Sci. 111 : 288-292.
by an excessive application of charcoal might be Graham, J. H. and L. Timmer. 1985. Rock phosphate as
concerned with an increment of soil pH value. a source of phosphorus for vesicular-arbuscular
mycorrhizal development and growth of citrus in
Therefore, an appropriate amount of charcoal to
a soilless medium. J. Amer. Soc. Hort. Sci. 110 :
be applied is less than 2 t per 10 a (this is approx- 489-492.
imately equivalent to 2% charcoal, Table 2). Graham, J. H., J. P. Syvertsen and M. L. Smith, Jr.
Furthermore, such an effect of charcoal may be 1987. Water relations of mycorrhizal and
strengthened by mixing charcoal and soil. phosphorus-fertilized non-mycorrhizal citrus under
VAM fungi develop well in citrus orchards drought stress. New Phytol. 105 : 411-419.
where Bahia grass is used for sod (Ishii et al., Harley, J. L. and E. L. Harley. 1987. A check-list of
1993). We have also indicated that the intensity of mycorrhiza in the British flora. New Phytol. 105 :
1-102.
VAM formation on some weeds grown in citrus
Ishii, T. and K. Kadoya. 1984. Growth of citrus trees
orchards was higher than that on citrus trees
as affected by ethylene evolved from organic mate-
(Ishii et al., 1989a). However, sod culture in com- rials applied to soil. J. Jap. Soc. Hort. Sci. 53 :
mercial citrus orchards has been unsuccessful in 320-330.
Japan; most citrus growers believe that a clean Ishii, T., K. Tatsumi and K. Kadoya. 1989a. VA
culture is best for the production of high-quality mycorrhizal development of citrus trees as
fruits. Thus, our soil management system must be affected by soil managements. J. Jap. Soc. Hort. Sci.
re-evaluated. 58 (Suppl. 1): 32-33. (In Japanese).
The prevailing cultural system in which large Ishii, T., K. Tatsumi and K. Kadoya. 1989b. Distribu-
tion and ecological aspects of vesicular-arbuscular
quantities of agrochemical and chemical fertilizers mycorrhizal fungi in citrus orchards. Mem. Coll.
are used, should be thoroughly revamped so that a
Agr., Ehime Univ. 34 : 65-71.
cultural system which maintains beneficial soil Ishii, T. and K. Kadoya. 1990. Use of charcoal as a
microorganisms is adopted. In conclusion, any ap- soil conditioner applied to citrus orchards. J. Jap.
plication of charcoal to the soil is a practical Soc. Hort. Sci. 59 (Suppl. 1): 36-37. (In Japanese).
method to improve soil properties and to foster the Ishii, T., Y. H. Shrestha and K. Kadoya. 1992a. VA
development of symbiotic microorganisms includ- mycorrhizal fungi in citrus soils and the relation-
ing VAM fungi. ship between soil factors and number of the
spores. J. Jap. Soc. Hort. Sci. 61 (Suppl. 2):
Literature Cited 166-167. (In Japanese).
Ishii, T., Y. H. Shrestha and K. Kadoya. 1992b. Effect
Antunes, V. and E. J. B. N. Cardoso. 1991. Growth and
of vesicular-arbuscular (VA) mycorrhizal fungi on
nutrient status of citrus plants as influenced by
tree growth, fruit development and quality, and
mycorriza and phosphorous application. Plant and
water stress of satsuma mandarin trees. J. Jap.
 J. Japan. Soc. Hort. Sci. 63(3) : 529-535. 1994. 535

Soc. Hort. Sci. 62 (Suppl. 1): 26-27. (In Japanese).
Ishii, T., J. Hamada, K. Ishizaki, Y. H. Shrestha and K.
Kadoya. 1993. Effect of sod culture system by
Bahia grass (Paspalum notatum Flugge.) on
vesicular-arbuscular mycorrhizal development of
satsuma mandarin trees. J. Jap. Soc. Hort. Sci. 62
(Suppl. 2): 98-99. (In Japanese).
Ishii, T. and K. Kadoya. 1993. Phytotoxic constituents
in the bark and sawdust extracts of Chamaecyparis
obtusa and Cryptomeria japonica and their effects on
the growth of seedlings of trifoliate orange (Ponci-
rus trifoliate Ref.) and rice (Oryza sativa L.). J. Jap.
Soc. Hort. Sci. 53 : 320-330.
Karnovsky, M. J. 1965. A formaldehyde-glutaralde-
hyde fixative of high osmolality for use in electron
microscopy. J. Cell Biol. 27 : 137A-138A.
Kobayashi, N. 1988. Factors affecting the germination
of spores of Gigaspara margarita . Microorganisms
31 : 13-28.
Krikun, J. and Y. Levy. 1980. Effect of vesicular
arbuscular mycorrhiza on citrus growth and
mineral composition. Phytoparasitica 8 : 195-200.
Menge, J. A., E. L. V. Johnson and R. G. Platt. 1978.
Mycorrhizal dependency of several citrus culti-
vars under three nutrient regimes. New Phytol. 81
: 553-559.
Miyazaki, S. 1697. Nougyou-Zensho. In : Yamada, T., J.
Iinuma, M. Oka and S. Morita (eds.). The complete
works of ancient agricultural books in Japan. Ru-
ral Culture Association, 1988. (In Japanese).
Nemec, S. 1979. Response of six citrus rootstocks to
three species of Glomus , a mycorrhizal fungus.
The citrus industry 5 : 5-14.
Ogawa, M. 1987. Mutualistic microorganisms at the
plant-soil interface. Rural Culture Association. (In
Japanese).
Phillips, J. M. and D. S. Hayman. 1970. Improved proc-
edures for clearing roots and staining parasitic
and vesicular-arbuscular mycorrhizal fungi for
rapid assessment of infection. Trans. Br. Mycol.
Soc. 55 : 158-161.
Shrestha, Y. H., T. Ishii and K. Kadoya. 1992. Effect of
vesicular-arbuscular (VA) mycorrhizal fungi on
photosynthesis and transpiration, and the dis-
tribution of photosynthates of fruit-bearing satsu-
ma mandarin trees. J. Jap. Soc. Hort. Sci. 62
(Suppl. 1): 28-29. (In Japanese).
Shrestha, Y. H., T. Ishii and K. Kadoya. 1993. A rela-
tion of vesicular-arbuscular mycorrhizal develop-
ment and fruit quality of satsuma mandarin. J. Jap.
Soc. Hort. Sci. 62 (Suppl. 2): 96-97. (In Japanese).
Tang, Z., Q. Zhang and S. Hou. 1984. The effects of
mycorrhizal fungus on phosphate uptake by citrus
in.red earth. Acta Mycologia Sinica 3 : 170-177.
Timmer, L. W. and R. F. Leyden. 1980. The relation-
ship of mycorrhizal infection to phosphorus-in-
duced copper deficiency in sour orange seedlings.
New Phytol. 85 : 15-23.

炭 施 用 が カ ンキ ツの樹 体 生 長 お よびVA菌 根 形 成 に及 ぼ す影 響

石 井 孝 昭1・ 門 屋 一 臣2

1愛 媛 大 学 教 育 学 部790愛 媛 県 松 山 市 文 京 町3

2愛 媛 大 学 農 学 部790愛 媛 県 松 山 市 樽 味3 -5-7

摘 要

炭 施 用 が カ ン キ ツ の 樹 体 生 長 お よ びVA菌 根形成に 根 形 成 は 対 照 区 よ り も炭 施 用 区 で 良 好 で あ り,特 にイ

及 ぼ す 影 響 を 調 査 した.イ ネ も み が ら,ベ イ ツ ガ樹 皮 ネ もみ が ら炭 で は そ の 感 染 率 が41.5%と 著 し く高 く,

あ る い は カ ンキ ツ ジ ュ ー ス か す か ら作 っ た 炭 で 処 理 し ま た 葉 内 の リ ン 含 量 も増 加 した.一 方,宮 内 イ ヨカ ン

た 土 壌 を 用 い て,ル ー トボ ッ ク ス に ウ ン シ ュ ウ ミ カ ン 園 に お け る炭(イ ネ も み が ら)施 用 区,バ ヒア グ ラス

(カ ラ タ チ 台)樹 を 移 植 し,こ れ にGlomusfascicula- 草 生 区,放 任 区 お よ び 慣 行 裸 地(除 草 剤 年3回 使 用)

の 胞 子 を 接 種 し た.そ の 結 果,ボ
tum ッ ク ス の ガ ラ ス 区 のVA菌 根 形 成 を 比 較 調 査 した と こ ろ,VA菌 根菌

面 に 観 察 さ れ る根 の 伸 長 は,い ず れの炭 施用 区 にお い の 感 染 率 は 炭 施 用 区(52.0%),バ ヒア グ ラス 草生 区

て も対 照(炭 無 施 用)区 よ り旺 盛 で あ っ た.全 生 体 重, (16.9%),放 任 区(7.3%),慣 行 裸 地 区(3.6%)の

地 下 部 重 お よ び新 梢 重 も炭 施 用 区 で 増 大 し た.VA菌 順 で あ っ た.
Microbial Fertilizers in Japan 6/25/09 7:07 AM

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Microbial Fertilizers in Japan :::

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Database Michinori Nishio

National Institute of Agro-Environmental Sciences
Annual Reports
Kannondai 3-1-1, Tsukuba, Ibaraki 305 Japan, 1996-10-01
Books
Newsletters This Bulletin discusses microbial products in Japan, where they are used on
Issues in Asian Agriculture many farms, particularly by organic farmers who hope that these products will
improve nutrient uptake by plants and the quality of their products. It
Extension Bulletins
discusses the use of charcoal and rhizobia to stimulate nutrient uptake, and
Technical Bulletins
the use of arbuscular mycorrizal fungi (AMF) to help establish vegetation on
Technical Notes barren land. The range of commercial AMF products available in Japan is
Practical Technology briefly described, and their use and effectiveness in Japanese agriculture.
Research Highlights
Abstracts in Other Languages: 中文, 日本語, !"#

Abstract

Introduction
In 1961, Japan enacted the "Fundamental Law of Agriculture", which
encouraged farmers to selectively produce vegetables, fruits, forage crops
and livestock as well as rice, instead of staple foods such as wheat, barley
and corn. The aim of the law was to raise farmers' incomes in response to the
rapid growth of the Japanese economy. Consumption of vegetables, fruits,
milk, eggs and meats increased with economic growth. Farmers adopted the
strategy of increasing crop yields by applying large amounts of chemical
fertilizers and pesticides. During the 1960s and 1970s, the yield of many
crops per unit area increased dramatically as the result of intensive use of
chemical inputs and various soil amendments.

At present, however, the yield of many crops in Japan has reached a plateau.
Moreover, the negative effects of heavy applications of chemical inputs are
becoming apparent, in terms of both production and the environment,
especially in the case of vegetables. Physiological disturbance of plant
metabolism is common, due to the accumulation of excess plant nutrients in
the soil. The spread of soil-borne diseases is a threat to vegetable production,
especially where monoculture is prevailing. Pollution of underground and
surface water by nitrates is sometimes reported from vegetable producing
areas. Quality deterioration, in terms of a decrease in the content of vitamins

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Microbial Fertilizers in Japan 6/25/09 7:07 AM

areas. Quality deterioration, in terms of a decrease in the content of vitamins

and sugars, is becoming a subject of concern. All these factors are giving
farmers an interest in the function and utilization of soil microorganisms, as a
way of repairing the damage from the overuse of chemical inputs.

Many farmers in Japan are showing a strong interest in the utilization of

microorganisms to help:
Stimulate plant nutrient uptake;
Provide biological control of soil-borne diseases;
Hasten the decomposition of straw and other organic wastes;
Improve soil structure; and
Promote the production of physio-logically active substances in the
rhizosphere or in organic matter.

The main incentive for farmers to use microorganisms seems to be that they
hope to increase the yield or quality of their crops at a relatively low cost,
without a large investment of money and labor. Although many microbial
materials are sold commercially, most of them are not microbiologically
defined, i.e. the microorganisms contained in the products are not identified,
and the microbial composition is not fixed. Many of these commercial
products are advertised as if they could solve any problem a farmer is likely
to encounter. Because most extension advisors lack any knowledge of
microbial products, confusion and trouble frequently occur.

In this report I would like to describe the present situation of microbial

technologies in Japan, focusing on the practical use of various products and
their potential.
Utilization of Arbuscular Mycorrhizal Fungi
More than 50% of upland and grassland soils in Japan are volcanic ash soils
(Andosols), which transform phosphate into unavailable forms by chemical
bonding with aluminum ions. Phosphate availability is therefore one of the
strongest limiting factors on Japanese upland and grassland farms. At present,
this problem is being overcome by a heavy basal dressing of a mixture of
superphosphate and fused phosphate. Although these heavy applications have
contributed to a remarkable increase in yields of many crops, many vegetable
fields have accumulated phosphate at levels which inhibit plant growth. On
the other hand, most grasslands are still deficient in phosphate, because
enough chemical phosphate is being applied only when they are reclaimed.
Therefore, there are two types of Andosols in Japan; one contains a sufficient
amount of phosphate, and one does not. In both cases, there have been
attempts to use arbuscular mycorrhizal fungi (AMF) or vesicular-arbuscular
mycorrhizal fungi (VAM) for soil amelioration.
Utilization of Indigenous Amf by the Application of Charcoal

The idea that the application of charcoal stimulates indigenous AMF in soil
and thus promotes plant growth is relatively well-known in Japan, although
the actual application of charcoal is limited due to its high cost. The concept
originated in the work of M. Ogawa, a former soil microbiologist in the
Forestry and Forest Products Research Institute in Tsukuba. He and his
colleagues applied charcoal around the roots of pine trees growing by the
seashore, and found that Japanese truffles became plentiful. He also tested
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Microbial Fertilizers in Japan 6/25/09 7:07 AM

seashore, and found that Japanese truffles became plentiful. He also tested
the application of charcoal to soybean with a small quantity of applied
fertilizer, and demonstrated the stimulation of plant growth and nodule
formation (Ogawa 1983). His findings with regard to legumes were taken up
for further study by the National Grassland Research Institute (Nishio and
Okano 1991).
Stimulation of Alfalfa Growth by Charcoal Application

Table 1 shows the results obtained with alfalfa in pot experiments. The soil
used was a volcanic ash soil with very low phosphate availability. Although
alfalfa growth was very poor without applied fertilizer, it was improved by the
application of small amounts of fertilizer, and even more by the application of
charcoal with the fertilizer.

Four sets of pots were prepared. Each set received the same amount of
fertilizer (2 g N, 4.4 g P and 8.3 g K/m2). Set [F] received only fertilizer. The
others received fertilizer and also rhizobia [F+R], 1,000g/m of charcoal
[F+C], and rhizobia plus charcoal [F+R+C]. The charcoal used was a
commercial product made of bark from several kinds of deciduous broad-
leaved trees. Particle composition was >2mm, 24%; 1-2mm, 18%, and
<1mm; 58%.

Compared to the sets which received fertilizer alone, or fertilizer plus rhizobia,
the charcoal application stimulated plant growth by 1.7 - 1.8 times [F+C] and
1.4 - 1.8 times [F+R+C], measured at 38 days after sowing. At this stage the
stimulatory effect of rhizobia on plant growth was not marked, because the
plants had met most of their requirements by absorbing the applied nitrogen
fertilizer, and nodule development was still at an early stage. At 58 days,
when the nitrate added had been completely exhausted, plants not inoculated
with rhizobia ([F] and [F + C]) ceased to grow, and their leaves turned yellow
due to nitrogen deficiency. The soil used did not contain any indigenous
rhizobia effective on alfalfa, so that roots not inoculated with R. meliloti did
not show any acetylene reduction activity. At this stage, the stimulatory effect
of charcoal on growth was observed only in the plants inoculated with
rhizobia. The shoot weight of the [F + R + C] plants was 1.7 times greater
than that of the [F + R] plants.
Stimulation of Nutrient Uptake by Charcoal Application

The amount of nutrients (N, P, K) absorbed by the shoots showed a trend

similar to that of the shoot fresh weight (Table 1). The amount of N fixed by
the nodules and transported to the shoots was calculated by subtracting the N
content of the shoots of the plants not inoculated with rhizobia from the N
content of the inoculated plants ([F+R]-[F], [F+R+C] - [F+C]). The addition
of charcoal increased this amount of N 2.8-4.0 times, and the ARA by 6.2
times (Table 2). Added charcoal also increased the nodule weight by 2.3
times.

Fig. 1 shows the relationship between the increment of P and N associated

with rhizobial inoculation in comparison with the non-inoculated alfalfa ([F+R]
- [F] and [+R+C] - [F+C]). A significant correlation was observed between
the increments of P and N, suggesting that the stimulation of nitrogen fixation
by charcoal addition may be due to the stimulation of P uptake.

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Microbial Fertilizers in Japan 6/25/09 7:07 AM

by charcoal addition may be due to the stimulation of P uptake.

Relationship between Charcoal Application and Amp

The relative values of the shoot fresh weight and the degree of AMF infection
were determined on the basis of the values of [F+R]. A significant correlation
was observed between the shoot weight and AMF infection (Fig. 2).

When the soil was sterilized by chloropicrin, alfalfa growth was greatly
reduced, even with the application of the same amount of fertilizer shown in
Table 1. The stimulatory effect of charcoal on plant growth also diminished.
On the other hand, vigorous plant growth and the stimulatory effects of
charcoal addition were clearly observed when the sterilized soil was mixed
with a large amount of native soil (Fig. 3). This clearly indicates that the
stimulatory effect of added charcoal may appear only when a certain level of
indigenous AMF are present.
Mechanism Whereby Charcoal Stimulates the
Growth of Amf
Charcoal may stimulate the growth of AMF by the following mechanism.
Charcoal particles have a large number of continuous pores with a diameter
of more than 100!m. They do not contain any organic nutrients, because of
the carbonization process. The large pores in the charcoal may offer a new
microhabitat to the AMF, which can obtain organic nutrients through mycelia
extended from roots. This may enable the AMF to extend their mycelia far out
from the roots, thus collecting a larger amount of available phosphate.
Utilization of Amf for Establishment of Green
Cover on Barren Land
Barren land with poor vegetation cover, such as bare slopes beside roads and
on mountains, or large fresh deposits of volcanic debris, are subject to serious
soil erosion. The ordinary method of establishing plant cover on sloping
barren land is to seed grass or transplant tree seedlings, together with
fertilization of the soil. At the early stages of plant development, however,
when plant cover is not yet well established, heavy rainfall can cause soil
erosion and leach out fertilizers. This retards plant establishment. To
overcome the problem, a new technology is now being developed.

T. Marumoto of Yamaguchi University and his colleagues have developed a

new soil mulching sheet made of a plastic random-fiber sheet of webbing. It
contains plant seeds, fertilizer (including a coated nitrogen fertilizer and
culture media composed of organic materials (peat soil + bark manure),
zeolite and bentonite (Marumoto 1996). The sheet is stretched out over the
soil surface, and helps prevent soil erosion at the early stages of vegetation
growth. Shoots and roots of seedlings can easily push through the sheet and
develop further. Marumoto also attempted to stimulate the growth of grasses
and trees by inoculating the sheet with AMF and ectomycorrhizal fungi. Table
3 shows one of their experiments, in which a commercial product of AMF
(Gigarospora margarita) was inoculated on the soil surface beneath a sheet
containing mixed grass seeds. After six months, the dry weight of the grass
increased by 1.4-1.6 times compared to the control, and the level of infection
by AMF was clearly enhanced by the inoculation.
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Microbial Fertilizers in Japan 6/25/09 7:07 AM

by AMF was clearly enhanced by the inoculation.

Marumoto et al. demonstrated the effectiveness of their sheet by applying it

to more than 50 field sites, including bare slopes of building construction
sites, sites where golf courses were being developed, road construction sites
and fresh deposits of volcanic debris. They are now attempting to improve
their technology by utilizing mycorrhizal fungi. Their experiments show that
the selection of plant species suited to the targeted soil, and also of species of
endo- and ecto-mycorrhizal fungi suited to the host plant, are very important.
Utilization of the Commercial Amf Products in
Vegetable Production
A number of AMF products for inoculation are sold commercially in Japan. In
May 1994, Idemitsu Industry, one of the biggest oil companies in Japan,
launched its AMF product (Yorifuji and Suzuki 1955). The Central Glass
Company then began to sell its AMF products through Tosho Central Trading
Company. Before these two companies, Kyowa Fermentation Industry had
been the first company in Japan to produce an AMF product and subject it to
marketing tests, but abandoned actual marketing since it judged there would
be little profit, in the context of Japanese intensive agriculture. Several other
companies are now investigating the application of AMF to agriculture, and
intend to market new products in the near future. As the sales of chemical
fertilizers fall, affected by the environmental conservation movement and by
the increasing costs of production, fertilizer companies are searching for
alternative added-value technologies, of which AMF is one. In addition to
stimulating the nutrient uptake by plants, it is hoped that AMF will prevent
the infection by pathogens of roots. If they are found in fact to do this, a very
large market demand might be expected, because soil-borne plant diseases
are the most serious limiting factor in Japan's vegetable production, where
continuous cropping is widespread. Since the microbiological industry
generally needs a relatively small investment, at least at the start, companies
other than fertilizer producers are also competing to develop and sell AMF
products.
Brief Description of Commercial Amf Products

Idemitsu Industry uses mainly strains of Glomus, with complementary strains

of Gigaspora and Scutellospora. Central Glass uses strains of Gigaspora.
Although the specificity of AMF is generally said not to be high, researchers at
these companies have demonstrated that different strains may sometimes
vary greatly in their ability to infect the roots of certain plants. Therefore,
AMF products are composed of multiple strains, all with confirmed infection
abilities. Spores and mycelia produced by the cultivation of host plants are
packed with mineral carriers or peat moss. One AMF product sold by Idemitsu
Industry has a water content of 15%, and should be stored at temperatures
lower than 20°C. Activity can be maintained for at least two years if the
product is stored at 5°C.
Effectiveness of Commercial Amf Products

AMF products are used mainly on vegetables such as eggplant, tomato,

strawberry, sweet pepper, leek, asparagus and lettuce. They are not used
much on flowers or fruit trees, although they probably will be in future. They

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much on flowers or fruit trees, although they probably will be in future. They
will probably not be used on low-value crops such as cereals, soybean and
pasture, because of the cost-benefit relationship.

In most cases, nursery beds are inoculated with AMF products, and the
inoculated seedlings are planted out in the field. Usually nursery beds also
receive 150-205 kg/ha of P2O5 fertilizer. Although this amount of phosphate
seems to be excessive in terms of producing full AMF activity, the growth of
seedlings has been reported to be increased significantly, by 20% or more, by
AMF inoculation. Final yields of marketable produce were reported to be
increased by 20-50%.

Vegetable farmers regard large, healthy seedlings as important in achieving

higher yields and preventing soilborne diseases. A more noticeable stimulation
of seedling growth could be expected if phosphate applications were reduced
to a lower level. Even so, farmers might not be satisfied if lower phosphate
applications resulted in smaller seedlings. In intensive agriculture, the
objective is not a reduction in chemical fertilizers, but a higher yield of better
quality. Organic farming using reduced inputs might be able to utilize AMF
products more effectively.
Utilization of Phosphate Solubilizing
Microorganisms
Japan has only very small amounts of rock phosphate, and most of its soils
immobilize phosphate ions into unavailable forms. Rock phosphate which can
be mined by current technology is predicted to become exhausted in about
100 years' time. Therefore, there is a strong interest in developing alternative
sources of phosphate fertilizer. Many countries are studying the direct
utilization of rock phosphate. Australia has developed "biosuper", i.e. pellets
composed of rock phosphate, sulfur and sulfur-oxidizing bacteria. Japanese
scientists are very interested in the solubilization of bound phosphate in soil
which has accumulated phosphate from repeated, heavy applications of
phosphate fertilizer.

While more than 70% of total phosphate is present in organic forms, such as
inositol phosphate in volcanic ash soils, there are very few indigenous
microorganisms with a strong ability to decompose inositol phosphate in the
soil. On the contrary, Japanese soils contain many indigenous heterotrophic
microorganisms which solubilize mineral bound phosphates by the excretion
of chelating organic acids. In grassland soils, phosphate solubilizing
microorganisms made up 1% of bacterial populations and 10% of fungal
populations (Nishio 1985). Tinker (1980) raised doubts on the utilization of
heterorophic phosphate solubilizing microorganisms, because they need a
large amount of organic matter before they can excrete organic acids. Even if
phosphate is solubilized, phosphate ions are incorporated into the microbial
biomass, so roots cannot absorb enough of them. Thus, we adopted the
following strategy: a) The addition of a large amount of organic matter makes
phosphate solubilizing (PS) microorganisms proliferate and these solubilize
bound phosphate. b) Solubilized phosphates are incorporated into the
microbial biomass during other microbial multiplication, using organic matter.
c) Once the organic matter becomes exhausted, the microbial biomass

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c) Once the organic matter becomes exhausted, the microbial biomass

decreases and releases phosphate into the soil. d) The death of the microbial
biomass can be accelerated by various soil treatments, including tillage,
drying, liming and sterilization. e) Plants can absorb phosphate after microbial
proliferation has ceased. f) The absorption of phosphate by plants can be
accelerated by inoculation with AMF.
Experimental Evidence

Each step described above has been experimentally confirmed (Kimura and
Nishio 1989). Fig. 4 shows the difference in biomass P (P retained in biomass)
between the soils with and without compound phosphate. When the soil was
incubated for 7 days by adding sucrose and ammonium sulfate, the biomass P
significantly increased, utilizing Ca-, Al-, and Fe-phosphates and low-quality
rock phosphate. However, no significant increase was observed with varicite
(crystallized aluminum phosphate) over this short period. This indicates that
insoluble phosphates which are not crystallized can be solubilized by
indigenous microorganisms when abundant carbon sources are supplied.

The rate of increase in soil biomass P fell, and available phosphate increased,
after the depletion of carbon sources, or after soil treatments such as
chloropicrin fumigation, air-drying or grinding (Fig. 5). This indicates that
after the exhaustion of organic matter, microbial biomass falls, releasing
phosphate into the soil, and that the release of available phosphate can be
accelerated by soil treatments.

To demonstrate the stimulatory effect of AMF on plant absorption of

phosphate released from soil biomass, an experiment was conducted using
dry yeast as an alternative to dead soil biomass. Fig. 6 shows that the
simultaneous addition of dry yeast and AMF had a marked effect on both the
growth of alfalfa and phosphate absorption by the plant.
Implications

Although this experimental evidence merely shows the principles underlying

the technology, this is very useful when we attempt to utilize heterotrophic
phosphate-solubilizing (PS) microorganisms in soil. It indicates that
PS microorganisms need the addition of a large amount of organic matter
as a substrate (excretes from roots are not sufficient);
Phosphate solubilized by PS micro-organisms is seldom absorbed directly by
the plant as long as a large amount of organic matter remains, because other
heterotrophs incorporate phosphate into biomass; thus,
Growth retardation of the plant may be a possibility just after the
application of organic matter.
To avoid growth retardation, seeding or transplanting should be delayed.
If rock phosphate, basic sludge or other low-grade phosphate is added, pre-
solubilization of bound phosphate in the compost is one way of avoiding
growth retardation.
Utilization of Microbial Materials in Organic
Farming
The number of farmers following organic farming is increasing each year in
Japan. The Ministry of Agriculture, Forestry and Fisheries adopted guidelines
for organic commodities in 1993. These define organic produce as being
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for organic commodities in 1993. These define organic produce as being

produced in fields to which no chemically synthesized inputs, except for those
permitted, have been applied for at least three years. Since crop production
with no chemical inputs at all is very difficult in Japan, many farmers instead
try to make minimal use of chemical fertilizer and pesticides. Produce grown
in this way is regarded as being related to organic food.

In terms of the plant nutrient supply, there are two types of organic farming.
One provides plant nutrients from local resources, and the other uses
commercial organic fertilizers. Most organic farmers in Japan use the latter
type, i.e. commercial organic fertilizers, made from rape seed meal or
soybean meal (both residues of oil extraction), meat and fish meal, bone
meal etc. These supply sufficient plant nutrients to give relatively high yields.
Local resources include green manures, composted livestock manure etc.
Utilization of Microbial Materials to Make
&Quot;Bokashi&Quot;

Most Japanese organic farmers utilize what is known as `bokashi', in addition

to compost. `Bokashi' is organic fertilizer which is briefly composted, to make
it less attractive to pests. If rape seed or soybean meal is directly applied to
soil, a certain fly ("tanebae") lays eggs in it. The maggots feed on young
seedlings just after germination and cause serious damage. Fishmeal also
attracts field mice, which dig tunnels under the seed beds. To avoid damage
of this kind, farmers developed on their own initiative a technique of
composting organic fertilizers for a short period. Typical ingredients are
shown in Table 4. These are mixed, and inoculated with microorganisms.
Water is added to give a moisture content of 50-55%, and the compost is
heaped into a pile. When the temperature of the pile reaches 50-55°C, the
pile is mixed and spread out. After the compost has cooled down, it is again
heaped in a pile. This microbial decomposition and cooling is repeated three
or four times. The materials are then spread out to dry, and finally packed in
bags for storage. The name of the product, `bokashi', means in Japanese
"obscuring the direct effectiveness". The concentration of nitrogen in bokashi
is much lower than in chemical fertilizer, ranging from 2 to 5% total nitrogen.

Since the original ingredients are dried materials, there are not enough
microorganisms present to begin immediate decomposition of the organic
matter. To avoid anaerobic fermentation, with its unpleasant odor, the
compost is inoculated with aerobic microorganisms which multiply rapidly.
Because these microorganisms need oxygen and have no heat-tolerance, the
pile is mixed and spread every one or two days. During the process of
composting, easily decomposable organic matter is decomposed through the
production of microbial biomass, liberating ammonium ions. The ammonium is
retained on soil particles. The microbial biomass contributes to the slower
release of nutrients with the residual ingredients. Overall, the aim of the
process seems to be, firstly, to decompose substances which attract pests,
and secondly, to create a slower-acting organic fertilizer.

The production of bokashi in Japan seems to be increasing. Many organic

farmers engaged in vegetable production use bokashi when they limit the
water supply to plants. Plants under water stress increase the osmotic
pressure in their fluid by increasing concentrations of mono- and oligo-
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pressure in their fluid by increasing concentrations of mono- and oligo-

saccharides instead of starch. This results in a higher level of sugars and
vitamins in the vegetables, as well as a longer post-harvest storage life.
These attributes raise the quality of the produce. When vegetables are being
cultivated in a state of water stress, the application of ordinary chemical
fertilizers is very difficult, because a rapid increase in the concentration of
mineral nutrients in a small amount of soil water frequently damages plant
growth. Although slow-acting fertilizers can avoid these difficulties, organic
farmers prefer bokashi because it is organic.

Although many microbial products are sold in Japan, except in a few special
cases, they are not microbiologically defined, i.e. the microorganisms they
contain are not identified, and merely described in terms of their hoped-for
results. Products which not only identify the microorganisms, but quantify
them, are very rare. Fortunately, the microorganisms effective in bokashi
production are not restricted to a special group, but are very common species
which can multiply rapidly in ordinary composting materials. No serious
problems have occurred in bokashi production, with one interesting exception.
Is &Quot;Em&Quot; Really an Effective Microorganism?
What is "EM"?

This exception is "EM", standing for "effective microorganisms". EM products

were developed by T. Higa of Ryukyu University, Okinawa. They contain
abundant anaerobic lactic acid bacteria and yeasts, as well as other
microorganisms. The utilization of these anaerobic microorganisms is a
distinctive feature which distinguishes EM from other microbial products. EM
first attracted notice in garbage treatment by local governments that were
struggling to cope with the increasing amount of garbage. The EM
manufacturer claimed that individual households could make "compost" of
good quality in one or two weeks using a sealed plastic bag or container
containing cooking refuse mixed with an EM product. Although anaerobic
fermentation usually generates an unpleasant odor, EM products were claimed
to suppress any bad smells by producing lactic acid. Higa claimed that the
"compost" thus prepared could be used in a home garden or distributed to
farmers. This idea attracted local governments, who hoped it would cut down
on the cost of garbage treatment, as well as citizens who appreciated the
importance of recycling. The "compost" thus prepared, however, has a very
high water content, because water vapor cannot escape from a sealed bag. It
also contains a large amount of available organic matter, because the
decomposition of organic matter is incomplete, as with the making of silage
or pickles. Incorporating available organic matter into the soil causes an
explosive proliferation of pathogenic "sugar fungi" such as Physium and
Rhizoctonia. Therefore, many crop failures have occurred when seeds were
sown just after application of the "compost". Some farmers' groups are now
making bokashi from this garbage compost by drying it, mixing it with other
materials, and composting this mixture further. "EM Bokashi"

Apart from compost made from household wastes, farmers are also making
bokashi, using EM products under anaerobic conditions, from other ordinary
organic materials. Higa claims in his book that a revolution in agriculture is
possible, since the use of EM products increases greatly the yield of crops. For

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possible, since the use of EM products increases greatly the yield of crops. For
example, rice fertilized with EM bokashi produced brown rice yields of more
than 12 mt/ha.

I. Goto and a colleague of Tokyo Agricultural University carried out some field
experiments in cooperation with the EM company. They analyzed EM bokashi
supplied by the company and also bokashi supplied by farmers, and showed
that the samples contained on average 40 kg N, 30 Kg P2O5 and 13 kg K2O
per 1000 kg, much the same as ordinary bokashi (Goto and Muramoto 1995).
EM recommends the application of 1000 kg/ha of EM bokashi, but the input of
40 kg of nitrogen contained in this amount is not sufficient for the full growth
of vegetables. Goto examined the crops of farmers who use EM bokashi, and
found that the yield of lettuce from 1000 kg of EM bokashi was much the
same as that from ordinary commercial organic fertilizer, when the nitrogen
levels were adjusted. He also found that farmers generally applied 30 mt/ha
of cattle manure in addition to the EM bokashi, and pointed out that the yields
obtained by the farmers may have been greatly supported by the manure.
When in fact vegetables were cultivated with only 1000 kg/ha EM bokashi
alone in the fields of the University, he found that the yields of many
vegetables were only half those obtained by ordinary practices (Muramoto
and Goto 1995). Later, researchers of EM reported that when the nitrogen
level was adjusted to 100 kg/ha, EM bokashi (2500 kg/ha) yielded the same
quantity of lettuce as chemical fertilizer (Iwahori et al. 1996). Therefore, they
insisted that the standard amount of EM bokashi which should be applied was
2500 kg/ha for lettuce, and criticized Goto for using an insufficient amount.
These experiments seem to show that EM bokashi is no revolutionary step
forward, because when it is used, vegetables need the same level of nitrogen
as when they are given chemical fertilizers. Goto claims that EM bokashi is
nothing but ordinary bokashi, and has no special qualities (Goto et al. 1996).
Utilization of Azolla in Organic Paddy Fields
Although Azolla has seldom been utilized by Japanese farmers, regardless of
experimental results in research institutes, the recent trend towards organic
farming shows signs of changing the situation. A common practice in organic
rice production is to release ducklings, usually hybrids of domestic ducks and
wild ones, into paddy fields. The webs of their feet disturb the soil surface in
the shallow water, and remove the young seedlings of weeds, thus controlling
weeds without herbicides. I. Watanabe, who studied Azolla at the
International Rice Research Institute, speculated that this soil disturbance
would suspend soil particles in the water, thus increasing the availability of
phosphate adsorbed on soil particles to Azolla floating on the water surface.
He made contact with a farmer and proposed his idea. They carried out
experiments in 1995 in the farmer's paddy field, inoculating Azolla in early
May without fertilizer. The Azolla increased very rapidly, and covered the
whole field just like a carpet. The Azolla biomass was estimated to be 28
mt/ha on June 25. Subsequently, some of the Azolla was eaten by birds, and
was reduced by about 30% by July 14. Since the Azolla supplied too much
nitrogen to the rice, some of the plants collapsed, and the yield of brown rice
was only 4.36-4.5 mt/ha (Huruno 1995). This experiment is likely to trigger
off widespread utilization of Azolla in Japan.

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Microbial Fertilizers in Japan :::

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Database Michinori Nishio

National Institute of Agro-Environmental Sciences
Annual Reports
Kannondai 3-1-1, Tsukuba, Ibaraki 305 Japan, 1996-10-01
Books
Newsletters This Bulletin discusses microbial products in Japan, where they are used on
Issues in Asian Agriculture many farms, particularly by organic farmers who hope that these products will
improve nutrient uptake by plants and the quality of their products. It
Extension Bulletins
discusses the use of charcoal and rhizobia to stimulate nutrient uptake, and
Technical Bulletins
the use of arbuscular mycorrizal fungi (AMF) to help establish vegetation on
Technical Notes barren land. The range of commercial AMF products available in Japan is
Practical Technology briefly described, and their use and effectiveness in Japanese agriculture.
Research Highlights
Abstracts in Other Languages: 中文, 日本語, !"#

Abstract

Introduction
In 1961, Japan enacted the "Fundamental Law of Agriculture", which
encouraged farmers to selectively produce vegetables, fruits, forage crops
and livestock as well as rice, instead of staple foods such as wheat, barley
and corn. The aim of the law was to raise farmers' incomes in response to the
rapid growth of the Japanese economy. Consumption of vegetables, fruits,
milk, eggs and meats increased with economic growth. Farmers adopted the
strategy of increasing crop yields by applying large amounts of chemical
fertilizers and pesticides. During the 1960s and 1970s, the yield of many
crops per unit area increased dramatically as the result of intensive use of
chemical inputs and various soil amendments.

At present, however, the yield of many crops in Japan has reached a plateau.
Moreover, the negative effects of heavy applications of chemical inputs are
becoming apparent, in terms of both production and the environment,
especially in the case of vegetables. Physiological disturbance of plant
metabolism is common, due to the accumulation of excess plant nutrients in
the soil. The spread of soil-borne diseases is a threat to vegetable production,
especially where monoculture is prevailing. Pollution of underground and
surface water by nitrates is sometimes reported from vegetable producing
areas. Quality deterioration, in terms of a decrease in the content of vitamins

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areas. Quality deterioration, in terms of a decrease in the content of vitamins

and sugars, is becoming a subject of concern. All these factors are giving
farmers an interest in the function and utilization of soil microorganisms, as a
way of repairing the damage from the overuse of chemical inputs.

Many farmers in Japan are showing a strong interest in the utilization of

microorganisms to help:
Stimulate plant nutrient uptake;
Provide biological control of soil-borne diseases;
Hasten the decomposition of straw and other organic wastes;
Improve soil structure; and
Promote the production of physio-logically active substances in the
rhizosphere or in organic matter.

The main incentive for farmers to use microorganisms seems to be that they
hope to increase the yield or quality of their crops at a relatively low cost,
without a large investment of money and labor. Although many microbial
materials are sold commercially, most of them are not microbiologically
defined, i.e. the microorganisms contained in the products are not identified,
and the microbial composition is not fixed. Many of these commercial
products are advertised as if they could solve any problem a farmer is likely
to encounter. Because most extension advisors lack any knowledge of
microbial products, confusion and trouble frequently occur.

In this report I would like to describe the present situation of microbial

technologies in Japan, focusing on the practical use of various products and
their potential.
Utilization of Arbuscular Mycorrhizal Fungi
More than 50% of upland and grassland soils in Japan are volcanic ash soils
(Andosols), which transform phosphate into unavailable forms by chemical
bonding with aluminum ions. Phosphate availability is therefore one of the
strongest limiting factors on Japanese upland and grassland farms. At present,
this problem is being overcome by a heavy basal dressing of a mixture of
superphosphate and fused phosphate. Although these heavy applications have
contributed to a remarkable increase in yields of many crops, many vegetable
fields have accumulated phosphate at levels which inhibit plant growth. On
the other hand, most grasslands are still deficient in phosphate, because
enough chemical phosphate is being applied only when they are reclaimed.
Therefore, there are two types of Andosols in Japan; one contains a sufficient
amount of phosphate, and one does not. In both cases, there have been
attempts to use arbuscular mycorrhizal fungi (AMF) or vesicular-arbuscular
mycorrhizal fungi (VAM) for soil amelioration.
Utilization of Indigenous Amf by the Application of Charcoal

The idea that the application of charcoal stimulates indigenous AMF in soil
and thus promotes plant growth is relatively well-known in Japan, although
the actual application of charcoal is limited due to its high cost. The concept
originated in the work of M. Ogawa, a former soil microbiologist in the
Forestry and Forest Products Research Institute in Tsukuba. He and his
colleagues applied charcoal around the roots of pine trees growing by the
seashore, and found that Japanese truffles became plentiful. He also tested
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seashore, and found that Japanese truffles became plentiful. He also tested
the application of charcoal to soybean with a small quantity of applied
fertilizer, and demonstrated the stimulation of plant growth and nodule
formation (Ogawa 1983). His findings with regard to legumes were taken up
for further study by the National Grassland Research Institute (Nishio and
Okano 1991).
Stimulation of Alfalfa Growth by Charcoal Application

Table 1 shows the results obtained with alfalfa in pot experiments. The soil
used was a volcanic ash soil with very low phosphate availability. Although
alfalfa growth was very poor without applied fertilizer, it was improved by the
application of small amounts of fertilizer, and even more by the application of
charcoal with the fertilizer.

Four sets of pots were prepared. Each set received the same amount of
fertilizer (2 g N, 4.4 g P and 8.3 g K/m2). Set [F] received only fertilizer. The
others received fertilizer and also rhizobia [F+R], 1,000g/m of charcoal
[F+C], and rhizobia plus charcoal [F+R+C]. The charcoal used was a
commercial product made of bark from several kinds of deciduous broad-
leaved trees. Particle composition was >2mm, 24%; 1-2mm, 18%, and
<1mm; 58%.

Compared to the sets which received fertilizer alone, or fertilizer plus rhizobia,
the charcoal application stimulated plant growth by 1.7 - 1.8 times [F+C] and
1.4 - 1.8 times [F+R+C], measured at 38 days after sowing. At this stage the
stimulatory effect of rhizobia on plant growth was not marked, because the
plants had met most of their requirements by absorbing the applied nitrogen
fertilizer, and nodule development was still at an early stage. At 58 days,
when the nitrate added had been completely exhausted, plants not inoculated
with rhizobia ([F] and [F + C]) ceased to grow, and their leaves turned yellow
due to nitrogen deficiency. The soil used did not contain any indigenous
rhizobia effective on alfalfa, so that roots not inoculated with R. meliloti did
not show any acetylene reduction activity. At this stage, the stimulatory effect
of charcoal on growth was observed only in the plants inoculated with
rhizobia. The shoot weight of the [F + R + C] plants was 1.7 times greater
than that of the [F + R] plants.
Stimulation of Nutrient Uptake by Charcoal Application

The amount of nutrients (N, P, K) absorbed by the shoots showed a trend

similar to that of the shoot fresh weight (Table 1). The amount of N fixed by
the nodules and transported to the shoots was calculated by subtracting the N
content of the shoots of the plants not inoculated with rhizobia from the N
content of the inoculated plants ([F+R]-[F], [F+R+C] - [F+C]). The addition
of charcoal increased this amount of N 2.8-4.0 times, and the ARA by 6.2
times (Table 2). Added charcoal also increased the nodule weight by 2.3
times.

Fig. 1 shows the relationship between the increment of P and N associated

with rhizobial inoculation in comparison with the non-inoculated alfalfa ([F+R]
- [F] and [+R+C] - [F+C]). A significant correlation was observed between
the increments of P and N, suggesting that the stimulation of nitrogen fixation
by charcoal addition may be due to the stimulation of P uptake.

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by charcoal addition may be due to the stimulation of P uptake.

Relationship between Charcoal Application and Amp

The relative values of the shoot fresh weight and the degree of AMF infection
were determined on the basis of the values of [F+R]. A significant correlation
was observed between the shoot weight and AMF infection (Fig. 2).

When the soil was sterilized by chloropicrin, alfalfa growth was greatly
reduced, even with the application of the same amount of fertilizer shown in
Table 1. The stimulatory effect of charcoal on plant growth also diminished.
On the other hand, vigorous plant growth and the stimulatory effects of
charcoal addition were clearly observed when the sterilized soil was mixed
with a large amount of native soil (Fig. 3). This clearly indicates that the
stimulatory effect of added charcoal may appear only when a certain level of
indigenous AMF are present.
Mechanism Whereby Charcoal Stimulates the
Growth of Amf
Charcoal may stimulate the growth of AMF by the following mechanism.
Charcoal particles have a large number of continuous pores with a diameter
of more than 100!m. They do not contain any organic nutrients, because of
the carbonization process. The large pores in the charcoal may offer a new
microhabitat to the AMF, which can obtain organic nutrients through mycelia
extended from roots. This may enable the AMF to extend their mycelia far out
from the roots, thus collecting a larger amount of available phosphate.
Utilization of Amf for Establishment of Green
Cover on Barren Land
Barren land with poor vegetation cover, such as bare slopes beside roads and
on mountains, or large fresh deposits of volcanic debris, are subject to serious
soil erosion. The ordinary method of establishing plant cover on sloping
barren land is to seed grass or transplant tree seedlings, together with
fertilization of the soil. At the early stages of plant development, however,
when plant cover is not yet well established, heavy rainfall can cause soil
erosion and leach out fertilizers. This retards plant establishment. To
overcome the problem, a new technology is now being developed.

T. Marumoto of Yamaguchi University and his colleagues have developed a

new soil mulching sheet made of a plastic random-fiber sheet of webbing. It
contains plant seeds, fertilizer (including a coated nitrogen fertilizer and
culture media composed of organic materials (peat soil + bark manure),
zeolite and bentonite (Marumoto 1996). The sheet is stretched out over the
soil surface, and helps prevent soil erosion at the early stages of vegetation
growth. Shoots and roots of seedlings can easily push through the sheet and
develop further. Marumoto also attempted to stimulate the growth of grasses
and trees by inoculating the sheet with AMF and ectomycorrhizal fungi. Table
3 shows one of their experiments, in which a commercial product of AMF
(Gigarospora margarita) was inoculated on the soil surface beneath a sheet
containing mixed grass seeds. After six months, the dry weight of the grass
increased by 1.4-1.6 times compared to the control, and the level of infection
by AMF was clearly enhanced by the inoculation.
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by AMF was clearly enhanced by the inoculation.

Marumoto et al. demonstrated the effectiveness of their sheet by applying it

to more than 50 field sites, including bare slopes of building construction
sites, sites where golf courses were being developed, road construction sites
and fresh deposits of volcanic debris. They are now attempting to improve
their technology by utilizing mycorrhizal fungi. Their experiments show that
the selection of plant species suited to the targeted soil, and also of species of
endo- and ecto-mycorrhizal fungi suited to the host plant, are very important.
Utilization of the Commercial Amf Products in
Vegetable Production
A number of AMF products for inoculation are sold commercially in Japan. In
May 1994, Idemitsu Industry, one of the biggest oil companies in Japan,
launched its AMF product (Yorifuji and Suzuki 1955). The Central Glass
Company then began to sell its AMF products through Tosho Central Trading
Company. Before these two companies, Kyowa Fermentation Industry had
been the first company in Japan to produce an AMF product and subject it to
marketing tests, but abandoned actual marketing since it judged there would
be little profit, in the context of Japanese intensive agriculture. Several other
companies are now investigating the application of AMF to agriculture, and
intend to market new products in the near future. As the sales of chemical
fertilizers fall, affected by the environmental conservation movement and by
the increasing costs of production, fertilizer companies are searching for
alternative added-value technologies, of which AMF is one. In addition to
stimulating the nutrient uptake by plants, it is hoped that AMF will prevent
the infection by pathogens of roots. If they are found in fact to do this, a very
large market demand might be expected, because soil-borne plant diseases
are the most serious limiting factor in Japan's vegetable production, where
continuous cropping is widespread. Since the microbiological industry
generally needs a relatively small investment, at least at the start, companies
other than fertilizer producers are also competing to develop and sell AMF
products.
Brief Description of Commercial Amf Products

Idemitsu Industry uses mainly strains of Glomus, with complementary strains

of Gigaspora and Scutellospora. Central Glass uses strains of Gigaspora.
Although the specificity of AMF is generally said not to be high, researchers at
these companies have demonstrated that different strains may sometimes
vary greatly in their ability to infect the roots of certain plants. Therefore,
AMF products are composed of multiple strains, all with confirmed infection
abilities. Spores and mycelia produced by the cultivation of host plants are
packed with mineral carriers or peat moss. One AMF product sold by Idemitsu
Industry has a water content of 15%, and should be stored at temperatures
lower than 20°C. Activity can be maintained for at least two years if the
product is stored at 5°C.
Effectiveness of Commercial Amf Products

AMF products are used mainly on vegetables such as eggplant, tomato,

strawberry, sweet pepper, leek, asparagus and lettuce. They are not used
much on flowers or fruit trees, although they probably will be in future. They

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Microbial Fertilizers in Japan 6/25/09 7:07 AM

much on flowers or fruit trees, although they probably will be in future. They
will probably not be used on low-value crops such as cereals, soybean and
pasture, because of the cost-benefit relationship.

In most cases, nursery beds are inoculated with AMF products, and the
inoculated seedlings are planted out in the field. Usually nursery beds also
receive 150-205 kg/ha of P2O5 fertilizer. Although this amount of phosphate
seems to be excessive in terms of producing full AMF activity, the growth of
seedlings has been reported to be increased significantly, by 20% or more, by
AMF inoculation. Final yields of marketable produce were reported to be
increased by 20-50%.

Vegetable farmers regard large, healthy seedlings as important in achieving

higher yields and preventing soilborne diseases. A more noticeable stimulation
of seedling growth could be expected if phosphate applications were reduced
to a lower level. Even so, farmers might not be satisfied if lower phosphate
applications resulted in smaller seedlings. In intensive agriculture, the
objective is not a reduction in chemical fertilizers, but a higher yield of better
quality. Organic farming using reduced inputs might be able to utilize AMF
products more effectively.
Utilization of Phosphate Solubilizing
Microorganisms
Japan has only very small amounts of rock phosphate, and most of its soils
immobilize phosphate ions into unavailable forms. Rock phosphate which can
be mined by current technology is predicted to become exhausted in about
100 years' time. Therefore, there is a strong interest in developing alternative
sources of phosphate fertilizer. Many countries are studying the direct
utilization of rock phosphate. Australia has developed "biosuper", i.e. pellets
composed of rock phosphate, sulfur and sulfur-oxidizing bacteria. Japanese
scientists are very interested in the solubilization of bound phosphate in soil
which has accumulated phosphate from repeated, heavy applications of
phosphate fertilizer.

While more than 70% of total phosphate is present in organic forms, such as
inositol phosphate in volcanic ash soils, there are very few indigenous
microorganisms with a strong ability to decompose inositol phosphate in the
soil. On the contrary, Japanese soils contain many indigenous heterotrophic
microorganisms which solubilize mineral bound phosphates by the excretion
of chelating organic acids. In grassland soils, phosphate solubilizing
microorganisms made up 1% of bacterial populations and 10% of fungal
populations (Nishio 1985). Tinker (1980) raised doubts on the utilization of
heterorophic phosphate solubilizing microorganisms, because they need a
large amount of organic matter before they can excrete organic acids. Even if
phosphate is solubilized, phosphate ions are incorporated into the microbial
biomass, so roots cannot absorb enough of them. Thus, we adopted the
following strategy: a) The addition of a large amount of organic matter makes
phosphate solubilizing (PS) microorganisms proliferate and these solubilize
bound phosphate. b) Solubilized phosphates are incorporated into the
microbial biomass during other microbial multiplication, using organic matter.
c) Once the organic matter becomes exhausted, the microbial biomass

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c) Once the organic matter becomes exhausted, the microbial biomass

decreases and releases phosphate into the soil. d) The death of the microbial
biomass can be accelerated by various soil treatments, including tillage,
drying, liming and sterilization. e) Plants can absorb phosphate after microbial
proliferation has ceased. f) The absorption of phosphate by plants can be
accelerated by inoculation with AMF.
Experimental Evidence

Each step described above has been experimentally confirmed (Kimura and
Nishio 1989). Fig. 4 shows the difference in biomass P (P retained in biomass)
between the soils with and without compound phosphate. When the soil was
incubated for 7 days by adding sucrose and ammonium sulfate, the biomass P
significantly increased, utilizing Ca-, Al-, and Fe-phosphates and low-quality
rock phosphate. However, no significant increase was observed with varicite
(crystallized aluminum phosphate) over this short period. This indicates that
insoluble phosphates which are not crystallized can be solubilized by
indigenous microorganisms when abundant carbon sources are supplied.

The rate of increase in soil biomass P fell, and available phosphate increased,
after the depletion of carbon sources, or after soil treatments such as
chloropicrin fumigation, air-drying or grinding (Fig. 5). This indicates that
after the exhaustion of organic matter, microbial biomass falls, releasing
phosphate into the soil, and that the release of available phosphate can be
accelerated by soil treatments.

To demonstrate the stimulatory effect of AMF on plant absorption of

phosphate released from soil biomass, an experiment was conducted using
dry yeast as an alternative to dead soil biomass. Fig. 6 shows that the
simultaneous addition of dry yeast and AMF had a marked effect on both the
growth of alfalfa and phosphate absorption by the plant.
Implications

Although this experimental evidence merely shows the principles underlying

the technology, this is very useful when we attempt to utilize heterotrophic
phosphate-solubilizing (PS) microorganisms in soil. It indicates that
PS microorganisms need the addition of a large amount of organic matter
as a substrate (excretes from roots are not sufficient);
Phosphate solubilized by PS micro-organisms is seldom absorbed directly by
the plant as long as a large amount of organic matter remains, because other
heterotrophs incorporate phosphate into biomass; thus,
Growth retardation of the plant may be a possibility just after the
application of organic matter.
To avoid growth retardation, seeding or transplanting should be delayed.
If rock phosphate, basic sludge or other low-grade phosphate is added, pre-
solubilization of bound phosphate in the compost is one way of avoiding
growth retardation.
Utilization of Microbial Materials in Organic
Farming
The number of farmers following organic farming is increasing each year in
Japan. The Ministry of Agriculture, Forestry and Fisheries adopted guidelines
for organic commodities in 1993. These define organic produce as being
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for organic commodities in 1993. These define organic produce as being

produced in fields to which no chemically synthesized inputs, except for those
permitted, have been applied for at least three years. Since crop production
with no chemical inputs at all is very difficult in Japan, many farmers instead
try to make minimal use of chemical fertilizer and pesticides. Produce grown
in this way is regarded as being related to organic food.

In terms of the plant nutrient supply, there are two types of organic farming.
One provides plant nutrients from local resources, and the other uses
commercial organic fertilizers. Most organic farmers in Japan use the latter
type, i.e. commercial organic fertilizers, made from rape seed meal or
soybean meal (both residues of oil extraction), meat and fish meal, bone
meal etc. These supply sufficient plant nutrients to give relatively high yields.
Local resources include green manures, composted livestock manure etc.
Utilization of Microbial Materials to Make
&Quot;Bokashi&Quot;

Most Japanese organic farmers utilize what is known as `bokashi', in addition

to compost. `Bokashi' is organic fertilizer which is briefly composted, to make
it less attractive to pests. If rape seed or soybean meal is directly applied to
soil, a certain fly ("tanebae") lays eggs in it. The maggots feed on young
seedlings just after germination and cause serious damage. Fishmeal also
attracts field mice, which dig tunnels under the seed beds. To avoid damage
of this kind, farmers developed on their own initiative a technique of
composting organic fertilizers for a short period. Typical ingredients are
shown in Table 4. These are mixed, and inoculated with microorganisms.
Water is added to give a moisture content of 50-55%, and the compost is
heaped into a pile. When the temperature of the pile reaches 50-55°C, the
pile is mixed and spread out. After the compost has cooled down, it is again
heaped in a pile. This microbial decomposition and cooling is repeated three
or four times. The materials are then spread out to dry, and finally packed in
bags for storage. The name of the product, `bokashi', means in Japanese
"obscuring the direct effectiveness". The concentration of nitrogen in bokashi
is much lower than in chemical fertilizer, ranging from 2 to 5% total nitrogen.

Since the original ingredients are dried materials, there are not enough
microorganisms present to begin immediate decomposition of the organic
matter. To avoid anaerobic fermentation, with its unpleasant odor, the
compost is inoculated with aerobic microorganisms which multiply rapidly.
Because these microorganisms need oxygen and have no heat-tolerance, the
pile is mixed and spread every one or two days. During the process of
composting, easily decomposable organic matter is decomposed through the
production of microbial biomass, liberating ammonium ions. The ammonium is
retained on soil particles. The microbial biomass contributes to the slower
release of nutrients with the residual ingredients. Overall, the aim of the
process seems to be, firstly, to decompose substances which attract pests,
and secondly, to create a slower-acting organic fertilizer.

The production of bokashi in Japan seems to be increasing. Many organic

farmers engaged in vegetable production use bokashi when they limit the
water supply to plants. Plants under water stress increase the osmotic
pressure in their fluid by increasing concentrations of mono- and oligo-
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Microbial Fertilizers in Japan 6/25/09 7:07 AM

pressure in their fluid by increasing concentrations of mono- and oligo-

saccharides instead of starch. This results in a higher level of sugars and
vitamins in the vegetables, as well as a longer post-harvest storage life.
These attributes raise the quality of the produce. When vegetables are being
cultivated in a state of water stress, the application of ordinary chemical
fertilizers is very difficult, because a rapid increase in the concentration of
mineral nutrients in a small amount of soil water frequently damages plant
growth. Although slow-acting fertilizers can avoid these difficulties, organic
farmers prefer bokashi because it is organic.

Although many microbial products are sold in Japan, except in a few special
cases, they are not microbiologically defined, i.e. the microorganisms they
contain are not identified, and merely described in terms of their hoped-for
results. Products which not only identify the microorganisms, but quantify
them, are very rare. Fortunately, the microorganisms effective in bokashi
production are not restricted to a special group, but are very common species
which can multiply rapidly in ordinary composting materials. No serious
problems have occurred in bokashi production, with one interesting exception.
Is &Quot;Em&Quot; Really an Effective Microorganism?
What is "EM"?

This exception is "EM", standing for "effective microorganisms". EM products

were developed by T. Higa of Ryukyu University, Okinawa. They contain
abundant anaerobic lactic acid bacteria and yeasts, as well as other
microorganisms. The utilization of these anaerobic microorganisms is a
distinctive feature which distinguishes EM from other microbial products. EM
first attracted notice in garbage treatment by local governments that were
struggling to cope with the increasing amount of garbage. The EM
manufacturer claimed that individual households could make "compost" of
good quality in one or two weeks using a sealed plastic bag or container
containing cooking refuse mixed with an EM product. Although anaerobic
fermentation usually generates an unpleasant odor, EM products were claimed
to suppress any bad smells by producing lactic acid. Higa claimed that the
"compost" thus prepared could be used in a home garden or distributed to
farmers. This idea attracted local governments, who hoped it would cut down
on the cost of garbage treatment, as well as citizens who appreciated the
importance of recycling. The "compost" thus prepared, however, has a very
high water content, because water vapor cannot escape from a sealed bag. It
also contains a large amount of available organic matter, because the
decomposition of organic matter is incomplete, as with the making of silage
or pickles. Incorporating available organic matter into the soil causes an
explosive proliferation of pathogenic "sugar fungi" such as Physium and
Rhizoctonia. Therefore, many crop failures have occurred when seeds were
sown just after application of the "compost". Some farmers' groups are now
making bokashi from this garbage compost by drying it, mixing it with other
materials, and composting this mixture further. "EM Bokashi"

Apart from compost made from household wastes, farmers are also making
bokashi, using EM products under anaerobic conditions, from other ordinary
organic materials. Higa claims in his book that a revolution in agriculture is
possible, since the use of EM products increases greatly the yield of crops. For

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Microbial Fertilizers in Japan 6/25/09 7:07 AM

possible, since the use of EM products increases greatly the yield of crops. For
example, rice fertilized with EM bokashi produced brown rice yields of more
than 12 mt/ha.

I. Goto and a colleague of Tokyo Agricultural University carried out some field
experiments in cooperation with the EM company. They analyzed EM bokashi
supplied by the company and also bokashi supplied by farmers, and showed
that the samples contained on average 40 kg N, 30 Kg P2O5 and 13 kg K2O
per 1000 kg, much the same as ordinary bokashi (Goto and Muramoto 1995).
EM recommends the application of 1000 kg/ha of EM bokashi, but the input of
40 kg of nitrogen contained in this amount is not sufficient for the full growth
of vegetables. Goto examined the crops of farmers who use EM bokashi, and
found that the yield of lettuce from 1000 kg of EM bokashi was much the
same as that from ordinary commercial organic fertilizer, when the nitrogen
levels were adjusted. He also found that farmers generally applied 30 mt/ha
of cattle manure in addition to the EM bokashi, and pointed out that the yields
obtained by the farmers may have been greatly supported by the manure.
When in fact vegetables were cultivated with only 1000 kg/ha EM bokashi
alone in the fields of the University, he found that the yields of many
vegetables were only half those obtained by ordinary practices (Muramoto
and Goto 1995). Later, researchers of EM reported that when the nitrogen
level was adjusted to 100 kg/ha, EM bokashi (2500 kg/ha) yielded the same
quantity of lettuce as chemical fertilizer (Iwahori et al. 1996). Therefore, they
insisted that the standard amount of EM bokashi which should be applied was
2500 kg/ha for lettuce, and criticized Goto for using an insufficient amount.
These experiments seem to show that EM bokashi is no revolutionary step
forward, because when it is used, vegetables need the same level of nitrogen
as when they are given chemical fertilizers. Goto claims that EM bokashi is
nothing but ordinary bokashi, and has no special qualities (Goto et al. 1996).
Utilization of Azolla in Organic Paddy Fields
Although Azolla has seldom been utilized by Japanese farmers, regardless of
experimental results in research institutes, the recent trend towards organic
farming shows signs of changing the situation. A common practice in organic
rice production is to release ducklings, usually hybrids of domestic ducks and
wild ones, into paddy fields. The webs of their feet disturb the soil surface in
the shallow water, and remove the young seedlings of weeds, thus controlling
weeds without herbicides. I. Watanabe, who studied Azolla at the
International Rice Research Institute, speculated that this soil disturbance
would suspend soil particles in the water, thus increasing the availability of
phosphate adsorbed on soil particles to Azolla floating on the water surface.
He made contact with a farmer and proposed his idea. They carried out
experiments in 1995 in the farmer's paddy field, inoculating Azolla in early
May without fertilizer. The Azolla increased very rapidly, and covered the
whole field just like a carpet. The Azolla biomass was estimated to be 28
mt/ha on June 25. Subsequently, some of the Azolla was eaten by birds, and
was reduced by about 30% by July 14. Since the Azolla supplied too much
nitrogen to the rice, some of the plants collapsed, and the yield of brown rice
was only 4.36-4.5 mt/ha (Huruno 1995). This experiment is likely to trigger
off widespread utilization of Azolla in Japan.

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 Plant and Soil 244: 273–279, 2002.
© 2002 Kluwer Academic Publishers. Printed in the Netherlands.
273

Inoculation with arbuscular mycorrhizal fungi: the status quo

in Japan and the future prospects

Masanori Saito1,3 & Takuya Marumoto2

1 Department ofGrassland Ecology, National Institute of Livestock and Grassland Science, Nishi-nasuno, Tochigi,
329-2793, Japan; 2 Faculty of Agriculture, Yamaguchi University, Yoshida, Yamaguchi, 753-8515, Japan;
3 Corresponding author∗

Received 21 August 2001. Accepted in revised form 12 December 2001

Key words: charcoal, degraded soil, inoculum, revegetation, VA mycorrhizal fungi

Abstract
Inoculation of arbuscular mycorrhizal (AM) fungi has potential benefits in not only sustainable crop production
but also environmental conservation. However, the difficulty of inoculum production due to the obligate biotrophic
nature of AM fungi has been the biggest obstacle to putting inoculation into practice. Nevertheless, several compan-
ies have sought to produce inoculum of AM fungi. Firstly in this review, the present status of inoculum production
and its use in Japan is described. Secondly, although the effectiveness of inoculation is primarily limited by environ-
mental and biological factors, some possible ways to improve inoculation performance are discussed. Approaches
include use of chemicals to increase spore germination and colonization and soil application of charcoal to provide
a microhabitat for AM fungi to colonize and survive.

Introduction der Heijden et al., 1998). Therefore, for rehabilitation

Inoculation with arbuscular mycorrhizal (AM) fungi
has potential value for improved crop production, and
numerous trials have been conducted since the 1970s
(Gianinazzi et al., 1990; Jarstfer and Sylvia, 1993,
1999; Kurle and Pfleger, 1994; Menge, 1984; Pow-
ell, 1984; Safir, 1994; Sieverding, 1991). However,
the difficulty in inoculum production due to the ob-
ligate biotrophic nature of AM fungi has been the
biggest obstacle to putting inoculation into practice.
At present, nevertheless, several companies all over
the world have commercialized the inoculum of AM
fungi (http://dmsylvia.ifas.ufl.edu/commercial.htm).
Sustainability of agricultural systems has been dis-
puted because of environmental pollution and deteri-
oration due to ‘modern’ and ‘intensive’ agriculture
(Matson et al., 1997). The significance of AM fungi to
the sustainability of the environment has been realized
in arable and natural lands because AM fungi affect
the plant community structure in various ways (Van
∗ FAX No: +81-287-36-6629. E-mail: msaito@affrc.go.jp
of deteriorated or degraded land, inoculation with AM
fungi can be considered an effective option (Miller and
Jastrow, 1992; Pfleger et al., 1994). Such a trend may
encourage the inoculum production companies.
The intention of this is not to review numerous
papers on inoculum production of AM fungi, but to
present the current status of inoculum production and
its use in Japan. Secondly, we discuss some pos-
sible ways to improve inoculation performance and the
future prospects in inoculation technology.
Inoculum production in Japan
In the 1940s in Japan, the growth promoting effect
of endomycorrhizal fungi on plant growth was estab-
lished by a pioneering scientist, T. Asai (Asai, 1943,
1944). However, his work was neglected until re-
search on inoculum production of AM fungi boomed
in the 1980s in Japan (Ogawa, 1987). In the early
1990s some companies started commercial production
of inoculum of AM fungi.
274
The Ministry of Agriculture, Forestry and Fisher-
ies (MAFF) of the Japanese Government has promoted
the introduction of various technologies to reduce
agrochemical inputs to arable lands for sustainable
agriculture. MAFF recognized that AM inocula are
useful to reduce phosphate fertilizer application, and
in 1996 approved AM inocula as soil amendments
by an ordinance of the Soil Productivity Improve-
ment Law. The ordinance regulates the quality of 12
types of amendments such as zeolite, peat, and other
organic/inorganic materials that are effective for the
improvement of soil productivity. The ordinance spe-
cifies that a quality guarantee be labeled on the outside
of the product container. In the case of AM fungi, the
following items are required; (i) name and address of
producer, (ii) raw materials, (iii) symbiotic efficiency,
(iv) efficacy, (v) recommended application rate, (vi)
storage conditions, (vii) expiration date. In item (ii),
carrier material, such as peat or zeolite, is indicated.
In item (iii), the symbiotic efficiency is expressed as
percentage of colonization by the inoculum of a spe-
cific test plant such as Welsh onion under standard
conditions. In item (iv), the producer should note that
the inoculum is not effective for some crops species
belonging to the Brassicaceae and Chenopodiaceae,
and that the inoculum may not be effective in soils
rich in available phosphate. This quality guarantee is
important to expel poor quality microbial inocula from
the market place.
At present, three companies, Central Glass Co.,
Idemitsu Kosan Co., and Osaka Gas Co., produce
AM inoculum. The MAFF statistics indicate that 28
– 83 tons of the inocula were supplied per year from
1997 to 1999. Much of inoculum was supplied for
non-agricultural applications such as rehabilitation of
degraded or devegetated landscapes. Because enduse
of the inocula is not within the framework of the or-
dinance, the above statistics do not include the supply
for such non-agricultural objectives.
Although the detailed procedure for inoculum pro-
duction is proprietary, these inocula are produced un-
der glasshouse conditions based upon the pot culture
technique (Jarstfer and Sylvia, 1993). One company
uses expanded clay as a potting medium (Dehne and
Backhaus, 1986). Others extract and concentrate the
propagules of AM fungi from potting media, and car-
riers such as peat are mixed with the propagules. The
inocula are mainly sold to horticulture farmers. Some
formulations are specific for each crop species by tak-
ing into account factors such as host plant species,
AM fungal species, and the handling of inoculum into
horticultural practices.
The cost of inoculum production is a serious prob-
lem because the inocula are not competitive in price
with phosphorus fertilizer. Even if farmers understand
the significance of sustainable agricultural systems,
the reduction of phosphorus inputs by using AM
fungal inocula alone cannot justify the use of the
inocula except in the case of high value crops. An-
other serious problem is control of phytopathogenic
micro-organisms. At present, the inoculum produced
is not completely free from pathogens, even though the
producers attempt to control pathogens with various
agrochemicals. Farmers are very aware of the risk of
pathogens, so they do not accept inoculum containing
host root residues. Although pieces of root colonized
with AM fungi, especially Glomus intraradices and
related species, function well as propagules, the com-
panies remove such residue of host roots from their
products.
Rehabilitation of a volcanic deposit-affected area
Inocula of AM fungi are expected to be substantially
beneficial in the establishment of vegetation in de-
graded or bare landscapes (Miller and Jastrow, 1992;
Pfleger et al., 1994; Requena et al., 2001). Currently
in Japan, AM fungal inocula has been applied most
successfully in revegetation of land by devastated by
volcanic activities (Marumoto et al., 1996, 1999).
In the Japanese islands, active volcanos sometimes
seriously damage large areas and destroy human activ-
ities. Newly deposited volcanic materials are very low
in nutrients available for plants and very susceptible to
erosion. The revegetation process is therefore slow, so
the acceleration of revegetation is required not only for
environmental conservation but also for the prevention
of erosion. In 1990, Mt. Fugendake (32◦ 45 N, 130◦
19 E) began volcanic activities after 200 years of in-
activity. During 1991 – 1994, several large pyroclastic
flows completely destroyed the previous vegetation on
the mountain slope. More than 1000 ha of the moun-
tain slope and the base of the mountain were covered
with a thick pyroclastic deposit.
Immediately after volcanic activities ceased in
1995, a revegetation project was started. Because the
area to be revegetated was on a steep slope of the vol-
cano and was still a hazard, the workers were not able
to access the revegetation project site during the first
a few years. Therefore, revegetation materials were
 275

Figure 1. Revegetation in a volcano-devastated area of Mt. Fugendake, Japan. (A) Application of bags containing AM fungal inoculum on
the pyroclastic flow. (B) Germination of plants from seeds contained in the bag. (C) A landscape of the pyroclastic flow 3 weeks after the
application. (D) A landscape in the same site as (C) 3 years after the application.

applied from the air by a helicopter in 1995. A bag of

unwoven polyester fabric, weighing about 2 kg, con-
tained plant seeds, AM fungal inocula, slow-release
chemical fertilizer, and some carriers such as peat
moss. Seeds of various wild grass and shrub species
were used: Miscanthus sinensis, Artemisia princeps,
Lespedeza cuneata and others. Gigaspora margarita
and Glomus sp. were used as the fungal inocula. About
3000 bags per ha were applied to the target-area (Fig-
ure 1 A, B). The grass plants that germinated from the
bag were highly colonized with AM fungi. Recently,
the inoculated fungal species were still proliferating 6
Figure 2. Schematic model of the effectiveness of AM fungal in-
years after application (unpublished). Thus, the AM oculation in relation to indigenous AM fungal potential and soil
fungi and some nutrients in the bag supported the phosphorus availability.
growth of the plants contained in the bag in the nutrient
poor pyroclastic flow. The site where the bags were
located became a base from which the plants reve- revegetation project, the cost for inoculum was not
getated the site and prevented serious erosion (Figure taken into account in the program.
1 C, D). This ‘bag’ method reduced the amount of
revegetation materials, including inoculum, and estab-
lished plant coverage more effectively than broadcast Improvement in inoculation performance
application of revegetation materials to the whole area.
Because the local government urgently requested the The effectiveness of AM fungal inoculation is affected
by various environmental and biological factors, espe-
276

cially the phosphorus availability in soil and the in-

oculum potential of indigenous AM fungi (Gianinazzi
et al., 1990). In soil low in phosphorus availability
and indigenous AM fungi, the effectiveness of inocu-
lation is expected to be the greatest (Figure 2). This
was demonstrated in the revegetation project described
above. Field trials in Japan with a commercial in-
oculum indicate that the efficacy is generally highest in
Andisols which show high phosphate fixing capacity
(Ueda and Kubo, pers. comm.). On the other hand, in
soils rich in available phosphorus, the effectiveness is
reduced low and the inoculation might even reduce the
crop performance (Peng et al., 1993).
Figure 3. Effect of temperature regime on spore germination of
Although the effectiveness of inoculation is Gigaspora margarita (modified from Miyamoto et al., 1994). The
primarily limited by inoculum potential and P avail- spores collected from a pot culture medium were mixed with cal-
ability, there are possible ways to improve inoculation cined attapulgite and were kept for 1 – 28 days at different temperat-
ures. After the treatment, 200 spores were extracted by wet-sieving,
performance. The inoculation process can be divided
and the germination rates for 2 weeks at 30 ◦ C were examined.
into three stages: (i) spore germination, (ii) colon-
ization, and (iii) growth of extraradical hyphae and
sporulation. The potential improvement of inoculum
performance is discussed for each stage of this pro-
cess.

Spore germination

The propagules (mainly spores) in the inoculum

should be active and should immediately initiate
growth after inoculation. Many chemical compounds
and unidentified fractions of various extracts have
been found to stimulate spore germination and hy-
phal extension (Azcón–Aguilar et al., 1999; Hepper,
1984; Ishii et al., 1997; Nagahashi and Douds, 2000;
Nair et al., 1991). Addition of these compounds to
the inoculum may improve the germination rate and
increase the colonization potential of the inoculum
(Tawaraya et al., 1998).
Spores of AM fungi often show dormancy, al-
though this phenomenon has not yet been critically
investigated. Storing the spores for a period from some
weeks to some months under cold temperature usually
breaks spore dormancy (Hepper, 1984; Safir et al.,
1990). G. margarita often shows spore dormancy and
a very low germination rate unless elevated temperat-
ure treatment is used to break the dormancy (Figure 3).
Breaking the spore dormancy technique can increase
initial colonization and may enhance the effectiveness
of the inoculation (Miyamoto et al., 1994).
Root colonization
Arbuscular mycorrhizal colonization is comprised
of a series of complicated processes from recogni-
Figure 4. Effect of charcoal application on the growth of alfalfa and
indigenous AM fungi in a pot experiment. Alfalfa was grown in a
clay loam infertile Andisol with N (2 g m−2 ) and K (8.3 g m−2 )
fertilization under different phosphate fertilizer application levels.
L, M and H in P level was equivalent to 0, 2.2 and 4.4 g P m−2 ,
respectively. Charcoal made of a mixture of barks from deciduous
trees was applied at a rate of 1 kg m−2 (modified from Nishio and
Okano, 1991).
tion between the fungi and host plant to arbuscule
formation (Nagahashi, 2000). Root exudates contain
compounds that promote colonization by AM fungi
(Nagahashi, 2000; Tawaraya et al., 1998). Since
the colonization process may depend on a balance
among phytohormones (Ludwig–Müller, 2000), and
some phytohormones affect colonization by AM fungi
(Ghachtouli et al., 1996). Application of a flavonoid,
formononetin at time of inoculation stimulated colon-
ization by AM fungi (Fries et al., 1998; Koide et al.,
1999). Application of such compounds at transplant-
ing may increase inoculation performance.

Growth of extraradical hyphae and sporulation
Nutrient absorption of mycorrhizal plants from soil
depends upon the function of extraradical hyphae. Soil
management that assures the extraradical hyphae to
be fully functioning may enable improvement of in-
oculation performance. Application of charcoal to soil
stimulated the colonization of crops by indigenous
AM fungi (Nishio and Okano, 1991; Ogawa et al.,
1983; Saito, 1990) (Figure 4). The effect of char-
coal was ascribed to its physioco-chemical properties.
Charcoal is porous, weakly alkaline, and does not
serve as a substrate for saprophytes. AM fungi sensit-
ive to competition from saprophytes can easily extend
their extraradical hyphae into charcoal buried in soil
and sporulate in the particles (Ogawa, 1987). Charcoal
particles act as a micro-habitat for AM fungi to survive
and later grow into the soil, which makes charcoal
suitable as a carrier of AM fungal inoculum (Ogawa,
1989). Charcoal can be applied in a large scale in in-
fertile soils rich in indigenous AM fungi, but its cost
is not competitive with inorganic fertilizers at least in
Japan.
Although many inoculation studies have been con-
ducted, there is still little known about how AM fungi
proliferate and survive in soil and how they affect
the following crop. In arable lands, various crops are
cultivated in sequence. Absence of a host plant, cul-
tivation of non-host crops, or long fallow negatively
affect the population of AM fungi in soil (Kurle and
Pfleger, 1994; Safir, 1994; Thompson, 1987). On
the other hand, some mycotrophic crops increase the
growth of the succeeding mycotrophic crop (Arihara
and Karasawa, 2000; Karasawa et al., 2001). Irrespect-
ive of inoculation, therefore, crop sequence should be
taken into account when predicting the responses of
AM fungi in a sustainable agricultural system.
Future prospects
Commercial inoculum production of AM fungi has
been increasing during the past decade, although
the future prospect of the business is still uncer-
tain. The obligate biotrophic nature of AM fungi
inevitably raises the cost for inoculum produc-
tion. Soilless culture systems seem to be prom-
ising, because this enables the production of clean
spores (Jarstfer and Sylvia, 1999). A dual culture
of transformed carrot hairy root with AM fungi
is an excellent system for providing the sterile
277
spores for research purposes (Glomales in vitro col-
lection; http://www.mbla.ucl.ac.be/ginco-bel/anglais
/cadre.htm) but is unsuitable for commercial inoculum
production. Even if mass production system of the
dual culture of lower production cost was developed
(Jolicoeur et al., 1999), it is uncertain whether the use
of spores produced from roots transformed by a patho-
genic bacterium, Agrobacterium rhizogenes, would be
allowed for inoculation in field.
In the field application of any microbial inoculum,
it is essential to verify that the inoculated micro-
organisms caused the plant response in field. Various
molecular techniques have been developed to dis-
tinguish the inoculated strain from other indigenous
strains. However, these techniques to identify AM
fungi are still not routinely used because of genetic
heterogeneity in AM fungi. Although ITS (internal
transcribed spacer) sequence of rDNA is widely used
for discrimination among strains in other fungal taxa,
application of the ITS sequence for identification of
AM fungi is complicated. For example, ITS sequences
of 3 isolates of G. margarita including BEG34 and
two Japanese isolates showed that the sequences were
too diverse to discriminate the isolates; the accession
number of the sequence data used were AJ006838–50
for isolates BEG34, AB048619–29 for MAFF520052,
and AB048607–18 for MAFF520054, respectively
(Saito, 2000). Single stranded conformation poly-
morphism (SSCP) of the large ribosomal subunit gene
may be promising to discriminate isolates of several
Glomus spp. (Kjøller and Rosendahl, 2000), while
the sequence diversity in this region is very high
(Clapp et al., 2001). We need more reliable molecu-
lar techniques to trace the inoculated fungi. Therefore,
from the standpoint of not only basic biological in-
terest but also application, molecular genetics of these
multinucleate fungi is of high research priority.
It is now well recognized that inoculation of AM
fungi has a potential significance in not only sustain-
able crop production, but also environmental conser-
vation. However, the status quo of inoculation is far
from practical technology that can be widely used in
the field. Together, a basic understanding of the bio-
logy of AM fungi and an improvement in inoculum
production and inoculation technology are required to
advance management of these fungi.
Acknowledgements
The preparation of this paper was supported in part by
278
Promotion of Basic Research Activities for Innovative
Biosciences (PROBRAIN), Bio-oriented Technology
Research Advancement Institution, Japan. The authors
are grateful to Central Glass Co. Ltd., Idemitsu Kosan
Co. Ltd., and Osaka Gas Co. Ltd. for valuable inform-
ation on inoculum production and to Prof. R. Koide
for his critical reading of this manuscript. MS is grate-
ful to Dr. Wu Chi–Guang for fruitful discussions on
inoculum production.
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 Soil Biology & Biochemistry 41 (2009) 1301–1310

Contents lists available at ScienceDirect

Soil Biology & Biochemistry

journal homepage: www.elsevier.com/locate/soilbio

Effect of biochar amendment on soil carbon balance and soil microbial activity
S. Steinbeiss a, *, G. Gleixner a, M. Antonietti b
a
Max Planck Institute for Biogeochemistry, Hans-Knoell-Str. 10, 07745 Jena, Germany
b
Max Planck Institute of Colloids and Interfaces, Am Muehlenberg 1, 14476 Potsdam-Golm, Germany

a r t i c l e i n f o a b s t r a c t

Article history: We investigated the behavior of biochars in arable and forest soil in a greenhouse experiment in order
Received 28 July 2008 to prove that these amendments can increase carbon storage in soils. Two qualities of biochar were
Received in revised form produced by hydrothermal pyrolysis from 13C labeled glucose (0% N) and yeast (5% N), respectively. We
16 March 2009
quantified respiratory losses of soil and biochar carbon and calculated mean residence times of the
Accepted 21 March 2009
Available online 17 April 2009
biochars using the isotopic label. Extraction of phospholipid fatty acids from soil at the beginning and
after 4 months of incubation was used to quantify changes in microbial biomass and to identify
microbial groups utilizing the biochars. Mean residence times varied between 4 and 29 years,
Keywords:
Biochar depending on soil type and quality of biochar. Yeast-derived biochar promoted fungi in the soil, while
13
C labeling glucose-derived biochar was utilized by Gram-negative bacteria. Our results suggest that residence
PLFA times of biochar in soils can be manipulated with the aim to ‘‘design’’ the best possible biochar for
Residence times a given soil type.
Greenhouse experiment Ó 2009 Elsevier Ltd. All rights reserved.

1. Introduction thousands of years (Pessenda et al., 2001; Gouveia et al., 2002;

There is a large imbalance between carbon release to the
atmosphere and carbon uptake by other compartments that leads
to a continued increase in atmospheric CO2 equivalent to a rate of
4.1 109 tons of carbon per year (IPCC, 2007). Thus, it should be of
utmost importance to develop new methods to retain carbon in
a stable form that can be stored outside of the atmosphere for
longer time periods. In this context, biochars have attracted a lot
of research within the last years basically with focus on the
application of biochars to soils, where they not only contribute to
carbon storage but at the same time act as fertilizers (Glaser et al.,
2001; Marris, 2006). Although a positive effect of biochar
amendments on crop yields was already known to ancient
cultures (Glaser, 2007), to date little is known about the effects of
biochar addition on soil microorganisms and consequently on the
soil carbon balance.
There is a huge variability in physical biochar structures
depending on the parent material and the conditions present at
their formation, which leads to quite different turnover times in
soils (Czimczik and Masiello, 2007). Large charcoal particles origi-
nated from forest wildfires have been shown to remain in soils for
* Corresponding author at: Institute of Groundwater Ecology, Helmholtz Centre
Munich, Ingolstaedter Landstr. 1, 85764 Neuherberg, Germany. Tel.: þ49 (0) 89 3187
2916; fax: þ49 (0) 89 3187 3361.
E-mail address: sibylle.steinbeiss@helmholtz-muenchen.de (S. Steinbeiss).
Gavin et al., 2003), however, smaller particles as derived from
grassland burning can hardly be detected in steppe ecosystems
(Forbes et al., 2006). The physical and chemical structure, e.g.
surface area and condensation grade, and the particle size of
synthetic biochars can be modified in technical processes (Titirici
et al., 2007a,b) opening the question about the stability of synthetic
biochars in soils.
There have been developed numerous chemical and technical
methods to produce charcoals from a variety of biomass materials
(Antal and Gronli, 2003; Marris, 2006; Titirici et al., 2007a). Each
production method needs a certain energy supply to activate the
reactions and results in completely different biochar structures.
However, hydrothermal carbonization looks especially promising
energy- and process-wise. Once activated in a continuous process,
20–30% of the energy bound to the original biomass are liberated
in the process, while keeping practically all carbon bound to the
final structure (Titirici et al., 2007b). No extensive biomass
material preparation or costly product isolation procedures are
required. Also soft, wet and low grade biomass can be carbonized,
making industrial biowaste, sludges or green household waste apt
to carbonization. A crude estimate of such directly accessible and
mostly already collected biowaste sums up to about 25  106 tons
per year in Germany, or to 10  109 tons per year worldwide.
Thus, we deal with a potential measure to cure at least significant
parts of the CO2 problem, appropriate biological stability in soils
and an added biological benefit provided. The optimal biochar

0038-0717/$ – see front matter Ó 2009 Elsevier Ltd. All rights reserved.
doi:10.1016/j.soilbio.2009.03.016
1302 S. Steinbeiss et al. / Soil Biology & Biochemistry 41 (2009) 1301–1310
combining fertilizer and carbon storage function in soils would
activate the microbial community leading to nutrient release and
fertilization and would add to the decadal soil carbon pool. The
structural and chemical properties of biochars that are driving
their decomposition or stabilization in soils still have to be
identified.
In our current study, we added two types of hydrothermally
synthesized biochar, a highly condensed, nitrogen free biochar
expected to be stable in soil, and a nitrogen containing biochar
with low condensation grade expected to be easily degradable,
to different soils with the aim to answer the following
questions:
1) How stable are biochars produced by this method in different
soils?
2) How do inherited soil microorganisms react on the addition of
such biochars?
3) Is the stability of these biochars tuneable by varying the
condensation grade and chemical composition of the biochar?
2. Materials and methods
2.1. Soil sampling and characterization
Soils used for the greenhouse experiment were sampled at the
continued arable plot of the Jena Experiment (Roscher et al., 2004)
and at the old growth forest field site of the Hainich National Park
(Knohl et al., 2003), respectively. The soil of the Jena Experiment
was classified as Eutric Fluvisol (FAO, 1998) and had a texture of 23%
clay, 64% silt and 13% sand (Kreutziger, personal communication).
The soil of the Hainich field site was a fertile Cambisol containing
40% clay, 56% silt and 4% sand (Knohl et al., 2003). In September
2007 the top 5 cm of soil were sampled at both field sites, passed
through a sieve with a mesh size of 2 mm and partitioned for PLFA
extraction (fresh soil), for soil column filling and for chemical
analyses (dried at 40  C), respectively.
Soil carbon and nitrogen concentrations were measured from
ball-milled sub-samples by elemental analysis (Elementar-
analysator vario Max CN, Elementar Analysensysteme GmbH,
Hanau, Germany) before and after incubation. Organic carbon
concentration was determined by calculating the difference
between elemental analyses of the total carbon concentration and
soil inorganic carbon concentration (Steinbeiss et al., 2008b). For
d13C analysis of the soil organic carbon, 3 mg ground sample was
weighed in small tin capsules. The arable soil contained about 1.6%
inorganic carbon, which was removed by treatment with 120 ml of
sulfurous acid (5–6% SO2, Merck, Darmstadt, Germany) prior to
isotope analysis (Steinbeiss et al., 2008a). Isotope ratios were
measured by a coupling of an elemental analyzer (EA 1110) with an
isotope ratio mass spectrometer (DeltaPlusXL, Thermo Finnigan,
Bremen, Germany). All values represent repeated measurements
with a standard deviation of less than 0.3& and were calibrated
versus V-PDB using CO2 as reference gas (Werner and Brand, 2001).
Soil analyses were summarized in Table 1.
Table 1
Basic characterization of biochars and soils; sd refers to standard deviation of replicated measurements. For d13C values of CO2 gas in the controls the standard error of the
calculated y-intercept was given instead (see Methods section for details).
Glucose-derived biochar
C content (%) 64.6
N content (%) 0.0
d13C value (&) 3.6
d13C value of CO2 gas in the controls (&)
2.2. Biochar production and characterization
Stable carbon isotopes have proven to reliably trace the flow of
carbon in various soil organic matter pools (Gleixner et al., 2001)
and into soil microorganisms using the compound-specific 13C
content of phospholipid fatty acids (Rubino et al., 2007; Kramer and
Gleixner, 2008). Moreover, changes in the13C content of various
pools enable the determination of mean residence times (Balesdent
and Mariotti, 1996; Gleixner et al., 2002). Consequently, the
production of isotopically labeled biochars from simple isotopic
precursors is most promising investigating synthetic biochars in
the soil system. Biochars were produced by hydrothermal pyrolysis
(Titirici et al., 2007a,b) using glucose (signature G) and yeast
(signature Y) as parent material, respectively. A 13C label was
introduced to both biochars adding uniformly 13C labeled glucose
(99 atom%, Sigma Aldrich, Seelze, Germany) to the parent materials
prior to biochar synthesis.
Glucose should be seen as model compound for cellulose, the major
structural component of plant biomass. Several investigations have
shown that charcoal produced from very different types of biomass
always show similar chemical structures (Schmidt and Noack, 2000;
Gleixner et al., 2001; Titirici et al., 2007b, in press). Heterocyclic (O-
containing) pyran and furan ring systems of carbohydrates or phenol
type structures that are the backbone of lignin form for example
benzene and other polyaromatic hydrocarbons (PAH) due to the
aromatization reactions in the charring process. Solid state 13C NMR
examinations proof this remarkable structural and compositional
similarity of all charcoals made from different sources of biomass
(Titirici et al., in press) and therefore we do not expect serious differ-
ences between charcoals produced from model compounds and from
biomass (Baccile et al., submitted for publication).
Yeast acted as a protein, i.e. nitrogen, rich model waste material
resulting from bioethanol, beer and wine production (pomace,
draff, brewer grains, distiller’s grain or distiller’s wash (Belyea et al.,
1998; Pfeffer et al., 2007; Maas et al., 2008; Quintero et al., 2008))
and thus represented a probable commercial source material for
the synthesis of nitrogen rich biochars. The yeast we used was
provided by a local beer brewery and represents the brewer grains
that were separated from the beer product as described in the
literature above. The grains were basically made up of the yeast
active in the fermentation and additionally contain some rest of
barley glume and wheat bran.
Element composition and d13C values of the biochars were
determined by elemental analysis and EA-IRMS as was described for
the soil samples, respectively (Table 1). Thermogravimetry
(TGA851e, Mettler-Toledo, Gießen, Germany) was applied to char-
acterize the thermal stability and thus the carbonization grade of the
biochars (Meszaros et al., 2007; Pastor-Villegas et al., 2007; Strezov
et al., 2007). Samples, biochars and their respective parent materials,
were introduced into the oven at 60  C and heated with a rate of
1  C min1 to 850  C in an Argon atmosphere. Scanning electron
microscopy (SEM) was performed on a DSM 940 A (Zeiss, Oberko-
chen, Germany). Infrared-spectra were measured with an IFS 66 FTIR
spectrometer (Bruker Optik GmbH, Ettlingen, Germany). Spectra
were obtained averaging 128 scans, with a resolution of 4 cm1.
sd Yeast-derived biochar sd Arable soil sd Forest soil sd
0.5 67.4 0.5 2.5 0.01 5.5 0.06
0.0 5.0 0.03 0.3 0.0003 0.5 0.005
0.15 2.8 0.30 27.7 0.14 27.1 0.06
24.0 0.8 27.6 0.6
 S. Steinbeiss et al. / Soil Biology & Biochemistry 41 (2009) 1301–1310
2.3. Experimental design and regular measurements
Soil columns were filled with 150 g soil (dry weight); 15
columns were filled with arable soil (signature A) and 15 columns
were filled with forest soil (signature F). The soil of six columns of
each soil type was mixed with glucose-derived biochar (signatures
AG and FG) and further six columns of each soil type were mixed
with yeast-derived biochar (signatures AY and FY). Three columns
of each soil type were left as control without biochar (signatures A
and F). The amount of biochar added to the soil was calculated to
correspond to a carbon addition of 30% of the initial soil organic
carbon content. Initial soil properties including PLFA analyses were
determined from soil samples without incubation (signatures AI
and FI) with triple replicates.
Soil columns were incubated at 25  C during the day and 20  C
during the night. No artificial lighting was applied. Soil moisture
was adjusted every three to four days in all columns. No water
leached out from the columns.
Soil respiration was measured using a carbon dioxide probe
(GMP 343, Vaisala, Helsinki, Finland) from week 1 to week 25 with
a temporal resolution of 1 week in the beginning (up to week 7) and
2–3 weeks afterwards. The respired gas was collected in weeks 7,
12, 15, 17, 20, 23 and 26 using 2.3 l gas flasks connected via
a capillary to the soil columns (filling time 4 h per sample). Sample
air was dried chemically using magnesium perchlorate (Fisher
Scientific, Loughborough, UK). Each time two flasks were filled the
same way with greenhouse air to correct ambient CO2 concentra-
tion and isotope ratios of the treatments (d13Ctreatment,korr)
(Amundson et al., 1998). Gas CO2 concentration was measured by
GC-FID (Agilent technologies, Santa Clara, USA) and stable carbon
isotope ratios were determined by isotope ratio mass spectrometry
(Finnigan MAT 252, Bremen, Germany).
Based on the isotopic difference between the respired CO2 from
biochars and from soil organic carbon it was possible to calculate
the proportion of biochar-derived carbon in the respired gas for
every sampling date (Equation (1)) (Balesdent et al., 1998; Waldrop
and Firestone, 2004). To overcome possible treatment effects we
determined the control values used in Equation (1) from the gas
measurements of the control treatments (A and F), assuming the
same processes in the treatments as in the controls. We made
Keeling plots combining the measured d13C values for the complete
time series of the gas collected from the respective control columns
and the reciprocal CO2 concentration (1/CO2). The y-intercept of the
linear fit (R2 ¼ 0.96 for A and R2 ¼ 0.97 for F) can be interpreted as
the d13C value of biologically produced CO2 during respiration
(Amundson et al., 1998) (Table 1).
d13 Ctreatment;corr  d13 Ccontrol
Fð%Þ ¼  100
d13 Cbiochar  d13 Ccontrol
Mean residence times (T) for the biochars were calculated using
measured carbon contents before (ct0) and after incubation (ct)
combined with the calculated proportion of biochar carbon in the
respiration gas assuming a first order reaction mechanism (Equa-
tion (2)) (Gregorich et al., 1996; Gleixner et al., 2002).
ðt  t0 Þ
T ¼
lnðct =ct0 Þ
2.4. Phospholipid fatty acid (PLFA) extraction
Phospholipid fatty acids were extracted from fresh sieved soil
before incubation and from all treatments after 4 months in the
greenhouse. We extracted three replicates of each treatment except
of the controls (signatures A, F). There, only one sample per soil
1303
type was extracted to leave the other two replicates for further
continuous measurements.
PLFA extraction was performed after standard methods
described in the literature (Bligh and Dyer, 1959; Zelles and Bai,
1993). Briefly, soils were shaken for 2 h in a mixture of chloroform,
methanol and phosphate buffer. The lipid extracts (chloroform
phase) were transferred to silica-filled solid phase extraction
columns (SPE). Phospholipids were separated from neutral lipids
and glycolipids by eluting with chloroform, acetone and methanol,
respectively. The phospholipids in the methanol fraction were
hydrolyzed and methylated using a methanolic KOH solution
leading to phospholipid fatty acid methyl esters.
Quantification of the PLFAs per soil dry weight was performed
on a GC-FID system (Agilent Technologies, Santa Clara, USA) using
a fused silica column (HP ultra 2, 50 m length  0.32 mm ID,
0.52 mm film thickness). The temperature program started at 140  C
(1 min isotherm) followed by a heating rate of 2  C min1 to 270  C,
which was held for 9 min, and followed by a final heating rate of
30  C min1 to 320  C. For peak identification retention times of
standard measurements were used.
Compound-specific isotope ratios of the identified PLFAs were
measured by GC–IRMS (DeltaPlusXL, Thermo Finnigan, Bremen,
Germany). Gas chromatographic separation was performed with
the same parameters as the quantification (see above). To obtain
d13C values of the PLFAs, measured isotope values were corrected
for the methyl carbon added during methylation (Kramer and
Gleixner, 2006).
Identified PLFAs were assigned to certain microbial groups, i.e.
fungi (C18:2u6,9, C18:1u9), Gram-positive bacteria (branched
saturated fatty acids), Gram-negative bacteria (monounsaturated
fatty acids) and bacteria in general (straight chain saturated fatty
acids) (Zelles, 1997; Baath and Anderson, 2003; Waldrop and
Firestone, 2004; Kramer and Gleixner, 2006; Allison et al., 2007).
2.5. Statistical evaluation
Statistical evaluation of the data sets was performed with SPSS
version 16.0 (SPSS Inc., Chicago, USA). For direct comparison of
treatments simple t-tests were used. To test for systematic effects of
the soil type, the charcoal type or any interaction of both variables
in complete data sets of all treatments analyses of variance
(ANOVA) were calculated. Principal component analyses using the
proportion of microbial groups in the soil were performed to
compare the structure of the microbial community in the different
treatments and the respective initial soil samples. Statistical
significance was assigned at the p  0.05 level.
3. Results
(1)
3.1. Biochar characterization
Glucose-derived biochar was highly carbonized and thus
thermally stable. Thermogravimetry to a temperature maximum
of 850  C in an inert atmosphere led to a total mass loss of about
50% with the highest rate of volatilization at temperatures
between 380 and 390  C. In contrast, the parent material, glucose,
(2) lost 86% of its initial mass under the same experimental condi-
tions and thermal degradation occurred in several processes with
maximum reaction rates already at temperatures of 200  C and
280  C, respectively.
The degree of condensation in the yeast-derived biochar was
much lower than that of the glucose-derived biochar, indicated by
a total mass loss of 72%, which is only 10% less than that of the
parent material, yeast, under the same conditions. Thermal
degradation of the yeast-derived biochar already started at 200  C,
1304 S. Steinbeiss et al. / Soil Biology & Biochemistry 41 (2009) 1301–1310

reached the next maximum reaction rate at 325  C and showed 1.1
a last process similar to the glucose-derived biochar degradation
between 380 and 390  C. 1.0
Scanning electron microscopy (SEM) showed similar structures
0.9
of our model biochars (carbonaceous spheres and continuous
C-H aromatic
nanopore systems) as were observed for charcoals produced from 0.8

Absorbance
different biomass types (e.g. rice grains, oak leafs, pine needles) by
hydrothermal pyrolysis (Cui et al., 2006; Titirici et al., 2007b) 0.7 C-O-C
(Fig. 1). CH2, CH3 aliphatic
Infrared-spectra reveal a polar surface structure of the model 0.6 CH
biochars containing phenolic, carbonyl and hydroxyl functional aliphatic C=O
0.5 C=C aromatic
groups. The aromatic structure was more pronounced for the highly
condensed glucose-derived biochar, whereas the low condensation
0.4
grade of the yeast-derived biochar resulted in larger proportions of
saturated and unsaturated aliphatic structures (Fig. 2). Infrared- 0.3 OH
spectra of charcoals produced from biomass verify hydroxyl groups,
phenolic residues, carbonyl functions, aliphatic double bonds and 4500 4000 3500 3000 2500 2000 1500 1000 500 0
a certain degree of aromaticity as the typical biochar structure Wavenumber in cm-1
characteristics (Titirici et al., 2007b).
Fig. 2. Infrared-spectra of glucose-derived biochar (straight line) and yeast-derived
biochar (dashed line).
3.2. Soil respiration and gas measurements

Initial respiration rates differed strongly between the treat-
ments (Fig. 3) but did not correlate to the initial carbon content.
In arable soil, both biochar treatments (AG, AY) showed similar
respiration rates to the control despite the carbon addition
(p ¼ 0.64 and 0.50, respectively). In forest soil, the highest initial
respiration was measured in the treatment with labile yeast-
derived biochar (FY), which was significantly higher (p < 0.001)
than both the control (F) and the treatment with glucose-derived
biochar (FG). No difference was observed between the treatment
with stable glucose-derived biochar in forest soil and the
respective control (p ¼ 1.00). Respiration rates strongly decreased
within 4 weeks of incubation and had leveled off after 12 weeks
to a constant median value of 2 mg C d1 for all treatments. No
systematic differences in respiration rates were observed between
the treatments, although the FY treatment showed significantly
higher respiration rates at some occasions within the first 10
weeks of the experiment (i.e. weeks 4 and 10: p < 0.001 and
p ¼ 0.062).
Labeling with 13C led to isotopic differences of 24–31&
between biochar carbon and soil organic carbon (Table 1) which
was used to quantify the proportion of biochar-derived carbon in
the respired CO2. Two major differences were observed between
the soil types. First, data variability as well between replicates as
between repeated samplings was much higher in arable soil
treatments than in forest soil treatments (p < 0.001). Second, the
proportion of biochar carbon in the respiration gas was generally
lower in forest soil treatments compared to arable soil treatments
(p ¼ 0.025). In detail, variability between replicates for arable soil
treatments were 5.4% (AG) and 3.3% (AY) on average, while
standard deviations of 0.9% were observed for replicates of both
forest soil treatments (FG and FY). The proportion of biochar-
derived carbon in the respiration gas was 28% (sd ¼ 7.9%) in the
AG treatment and 22% (sd ¼ 24.3%) in the AY treatment. In forest
soil 8% (sd ¼ 1.2%) of the respired carbon derived from biochar in
the FG treatment and 12% (sd ¼ 3.6%) derived from biochar in the
FY treatment. In the AG treatment, the proportion of biochar-
derived CO2 showed an increasing trend with time, reaching the
maximum of 43% in week 20. The AY treatment showed the most
inconsistent pattern in the composition of the respiration gas
during incubation, varying between 0 and 53% biochar carbon in
the gas without any regularity. In contrast, in the forest soil
treatments the proportion of biochar carbon in the respiration gas

Fig. 1. SEM picture of (a) glucose-derived biochar and (b) yeast-derived biochar. Scale bar 10 mm.
 S. Steinbeiss et al. / Soil Biology & Biochemistry 41 (2009) 1301–1310 1305

A F
60 AG 60 FG
AY FY

50 50
respiration rate in mg C d-1

40 40

30 30

20 20

10 10

0 0
0 5 10 15 20 25 0 5 10 15 20 25
week after experiment start week after experiment start

Fig. 3. Respiration rates for arable soil treatments (left) and forest soil treatments (right) including the respective controls during incubation. Error bars represent standard
deviations between three replicates per treatment.

was constant at 8% for FG, while FY showed a decreasing trend
from 19% in week 7 down to 9% in week 17 and later on.
3.3. Development of soil carbon stocks and mean residence times
Measured respiration rates reflect temporary carbon losses and
varied over time due to temperature and soil moisture variability.
The total carbon budget was thus determined by elemental analysis
from soil sub-samples before and after four months of incubation
(Table 2). Total carbon content decreased in controls and all treat-
ments over the incubation time. Despite different absolute
numbers in the carbon budget due to the different initial carbon
content of the two soil types, the relative amount of carbon
remaining in the soils after biochar incubation only depended on
the type of biochar added but did not depend on the soil type itself
or any interaction between soil type and biochar type (Table 3).
Thus, of the initially added 3% of charcoal carbon, both treatments
with more stable glucose-derived biochar still contained 27% more
carbon than the respective control, while both treatments with
labile yeast-derived biochar still contained 23% more carbon than
the respective control (Table 2).
To quantify losses of biochar carbon and soil organic carbon
we used the isotopic signature of the respiration gas. We
normalized all losses to the respective initial carbon amounts of
Table 2
Carbon budget in g C and relative to the respective controls for all treatments before
and after incubation; sd refers to standard deviation of replicated measurements.
the treatments for better comparability (Fig. 4). Losses of soil
organic carbon were generally smaller in the arable soil treat-
ments compared to the respective forest soil treatments and
biochar addition always increased the loss of carbon from the soil
organic carbon pool (Table 4). In both soil types, soil organic
carbon losses were largest when labile yeast-derived biochar was
added (Fig. 4, Table 4). In these treatments, twice the amount of
soil organic carbon was respired compared to the controls.
Moreover, yeast-derived biochar seemed to be better degradable
than glucose-derived biochar as indicated by larger biochar
carbon losses in both soil types (Fig. 4), although the difference
was significant only in forest soil (p ¼ 0.003, p ¼ 0.147 for arable
soil). Most interestingly, normalized losses of soil and biochar
carbon in the arable soil were almost identical (p ¼ 0.544), while
in forest soil less biochar carbon than soil organic carbon was lost
(p < 0.001).
The mean residence times (Equation (2)) for biochar carbon
were calculated from measured total carbon losses and the
proportions of biochar carbon in the respiration gas (Equation
(1)). In total, the controls lost 4% (A) and 6% (F) of their initial
carbon content, treatments with glucose-derived biochar lost 7%
(AG) and 8% (FG) carbon, and addition of yeast-derived biochar
caused total carbon losses of 10% (AY) and 11% (FY), respectively.
Consequently, mean residence times ranged between 4 (AG) and
29 (AY) years (Fig. 5). The 29 years were calculated for yeast-
derived biochar in arable soil and give just a rough estimate due
to the huge uncertainty caused by the high variability in the
detected proportion of biochar carbon in the respiration gas.

Treatment Cinitial sd Amount C relative Cfinal sd Amount C relative Table 3

(g) to control (%) (g) to control (%) Summary of analysis of variance (ANOVA) of the carbon budget relative to the
A 3.79 0.02 100 3.63 0.02 100 respective controls for all biochar treatments after incubation.
AG 4.93 0.02 130 4.61 0.03 127
AY 4.93 0.02 130 4.45 0.04 123 Parameter Sum of squares F-value Significance
F 8.23 0.04 100 7.75 0.04 100 Soil 0.80 1.07 0.332
FG 10.70 0.05 130 9.87 0.01 127 Charcoal 56.77 75.52 <0.001
FY 10.70 0.05 130 9.53 0.06 123 Soil  charcoal 0.01 0.01 0.923
1306 S. Steinbeiss et al. / Soil Biology & Biochemistry 41 (2009) 1301–1310

treatment 90
A AG AY F FG FY
0 80

mean residence time in years

-2 70
-4
60
-6
% carbon lost

-8 50
-10
40
-12
-14 30

-16 20
-18
10
-20 soil organic carbon
-22 biochar carbon 0
AG AY FG FY
Fig. 4. Losses of biochar carbon and soil organic carbon after 4 months of incubation treatment
given relative to the respective initial amounts in the treatments. Error bars reflect the
uncertainties in the proportion of biochar carbon in the respiration gas used for Fig. 5. Calculated mean residence times for the biochars in the different treatments.
calculation and were determined according to error propagation laws. Error bars reflect the uncertainty caused by the variability of the proportion of biochar
carbon in the respiration gas. They were determined according to error propagation
laws.

Glucose-derived biochar remained longer in forest soil (12

years) than in arable soil (p < 0.001), where it would be
mineralized after 4 years, assuming a continuously ongoing
quite similar (Fig. 6). Again, major effects of biochar addition
decomposition as observed in the 4 months of incubation. For
were the same in both soil types and depended only on the
yeast-derived biochar it would take about 6 years to be miner-
biochar type (Fig. 6, Table 6). While the addition of glucose-
alized in the forest soil.
derived biochar rarely changed the composition of the soil
microbial community, the yeast-derived biochar strongly
promoted fungi in both soils (p < 0.001). The majority of present
3.4. Microbial community in the soils
microorganisms belonged to the group of usually root associated
Gram-negative bacteria and made up 42% (AI) and 44% (FI) of the
To get an estimate of microbial adaptation to the new carbon
microbial biomass. In both soils, 27% of the microbes belonged to
source the total amount of phospholipid fatty acids in the soils was
the Gram-positive bacteria, 11% were assigned to bacteria in
determined before and after incubation in all treatments. As
general and only 11% (FI) to 12% (AI) of the microbial community
already found for the total carbon budget, biochar type was the
was made up of fungal biomass. The addition of glucose-derived
driving parameter for any effects on the microbial community
biochar led to shifts along PC 2 in the principal component
between the treatments. Arable soil generally contained smaller
analyses (PCA) (Fig. 6), which only explained 4–11% of the
amounts of microorganisms than forest soil as well for the initial
variance and reflected non-systematic changes in the proportion
values as in every treatment after incubation (p < 0.001). Addition
of all microbial groups. In treatments with yeast-derived biochar,
of glucose-derived biochar to both soil types caused a significant
the proportion of fungal biomass increased by 16% in both soil
reduction of microbial biomass (p < 0.001) during incubation. In
types, while Gram-positive and Gram-negative bacteria
contrast, yeast-derived biochar addition did not change the PLFA
decreased by 7–14%. The increase in fungal biomass was
content in the soils (p ¼ 0.39), which still was as high as before
expressed in the large shifts along PC 1 of the PCA (p < 0.001)
incubation in both soil types. We found no interaction between soil
(Fig. 6), which explained 89% of the variance in forest soil and
type and biochar type (p ¼ 0.15).
94% in arable soil. The microbial community composition of the
The identified PLFAs were assigned to four main groups of
controls without biochar addition showed the same pattern after
microorganisms in soil, i.e. fungi, Gram-positive bacteria, Gram-
incubation as the initial soil samples (Table 5).
negative bacteria and bacteria in general. The proportions of
these groups in the microbial community were calculated for the
initial soils and for all treatments after incubation (Table 5). The
initial microbial community composition in both soil types was Table 5
Proportion of the amount of PLFAs assigned to different microbial groups, i.e. fungi,
Gram-negative bacteria, Gram-positive bacteria and bacteria in general, before and
after incubation; sd refers to standard deviation between three replicates. There was
Table 4 no replicate extracted for the controls (A, F).
p-Values resulting from multiple comparison t-tests of soil organic carbon losses
(normalized to the initial carbon content) for all treatments after incubation. treatment Fungi sd Gram() sd Gram(þ) sd Bacteria sd
Numbers smaller than 0.05 reflect significant differences between the treatments. bacteria bacteria
AI 11.9 0.1 41.9 0.1 26.9 0.1 11.5 0.1
A AG AY F FG FY AG 10.8 0.3 41.8 2.1 25.0 2.4 13.0 0.3
A 0.120 0.031 AY 28.0 0.9 30.7 0.4 18.8 0.5 15.7 0.2
AG 0.120 0.004 0.001 FI 11.0 0.2 43.6 0.5 26.8 0.4 10.8 0.0
AY 0.031 0.004 0.018 FG 10.7 0.2 37.7 0.3 29.9 0.4 13.8 0.1
F 0.005 0.013 FY 27.6 0.8 28.9 0.3 20.3 0.7 16.8 0.1
FG 0.001 0.005 0.001 A 12.0 41.9 26.9 11.6
FY 0.018 0.013 0.001 F 10.9 39.0 27.3 14.5
 S. Steinbeiss et al. / Soil Biology & Biochemistry 41 (2009) 1301–1310 1307

PC 2 Assuming biochar and soil organic carbon as only carbon

2,5 sources for the soil microorganisms, the proportion of biochar
carbon incorporated in the microbial biomass can be calculated.
2,0 For instance, the observed enrichment in fungal biomarkers of
14.8& in the AY treatment would equal a utilization of 60%
1,5 biochar carbon for fungal biomass production. In forest soil,
fungi used 40% yeast-derived biochar carbon to build up their
FG
1,0 biomass. Bacteria in arable soil used between 10% and 15%
AG glucose-derived biochar carbon as carbon source and the
0,5 proportion of glucose-derived biochar carbon in Gram-negative
-1,5 -1,0 -0,5 0,5 1,0 1,5 2,0 2,5 bacteria in forest soil amounted to 13%.
PC 1
FY AY 4. Discussion
-0,5
AI
It has been observed in several studies that biochar addition to
-1,0
FI soils improved soil fertility and thus increased crop yields on
-1,5
agricultural lands (Marris, 2006; Chan et al., 2007). This fertilizer
effect could be explained by a stimulation of soil microorganisms
Fig. 6. Principal component analyses of the microbial community composition for both that consequently led to an increased recycling of nutrients
soil types. PC 1 explained 89% (forest soil) and 94% (arable soil) of the variance and was trapped in biomass residues. The fertilizer function is additionally
driven by the proportion of fungi and bacteria in both soil types. PC 2 explained 11%
supported by an increased water retention and cation exchange
(forest soil) and 4% (arable soil) of the variance.
capacity of the soils caused by the huge surface area of the bio-
chars. An aspect of biochar amendments that got more attention
Finally, we determined compound-specific isotope ratios of recently is that additional photosynthetically fixed carbon is
the PLFAs to identify the major carbon sources of the different brought into the soil, where it could contribute to longer term
groups of soil microorganisms (Appendix 1). PLFA enriched in carbon storage and thus mitigates increasing atmospheric CO2
13
C compared to the initial value indicated the uptake of biochar concentrations (Schmidt and Noack, 2000; Lehmann, 2007).
carbon by the respective microbial group, while unchanged d13C However, little is known about turnover times of biochars in soils
values prove the continuous uptake of soil organic carbon. and a long-term storage function contradicts the fertilizer func-
Control treatments generally showed small shifts in all microbial tion of biochars that requires a certain biodegradability of the
groups as decomposition of soil organic carbon led to an biochar material. The major question to solve will be to design
enrichment in 13C in the remaining soil organic carbon. Although biochars that fulfil both functions with the best possible
the microbial group specific enrichments somehow depended on compromise. Biochar structure, e.g. the condensation grade, could
the type of biochar added to the soils, e.g. significant enrich- easily be managed in production processes, but studies are
ments of Gram-negative and Gram-positive bacteria for glucose- necessary to check, whether the condensation grade is a tool to
derived biochar treatments (2.1–4.7&, 0.001 < p < 0.027) and control the turnover of biochars in soils.
strong yeast-derived biochar uptake by fungi in both soils In our greenhouse experiment we investigated the conse-
(isotopic shift 9.5 and 14.8&, 0.003 < p < 0.062), several differ- quences of the addition of biochar with different condensation
ences were observed between the soil types (Fig. 7). In contrast grades (high ¼ glucose-derived, low ¼ yeast-derived) to two soil
to arable soil, where fungi were less involved in the decompo- types (arable and forest soil) for inherited carbon stocks and for soil
sition of glucose-derived biochar (isotopic shift ¼ 1.6&, microbial communities.
p ¼ 0.350), in forest soil fungal biomarkers showed an average Soil respiration measurements indicated a strong stimulation
enrichment of 4.8& (p ¼ 0.417). Especially the d13C value of of soil microorganisms by yeast-derived biochar in the beginning
C18:2u6,9 increased by 8.4& (p ¼ 0.007), while other fungi of the incubation, whereas treatments that received glucose-
biomarkers (C18:1u9) remained unchanged (isotopic derived biochar showed respiration rates similar to the respec-
shift ¼ 1.1&). Beside the obvious utilization of yeast-derived tive controls. After 12 weeks no differences between any
biochar by fungi in arable soil, also all bacterial groups were able treatments could be observed anymore and respiration rates had
to decompose this biochar to a certain extent in this soil type. generally decreased to very low levels. As a result of the
Significant isotopic enrichments were measured in all bacterial increased respiration total carbon losses were always higher in
biomarkers (p  0.002). In forest soil, Gram-negative bacteria treatments that received biochar than in the controls. The type
were the only microbial group beside fungi that took up yeast- of biochar clearly showed a systematic influence on soil organic
derived biochar carbon in a significant amount (p ¼ 0.001). carbon losses. Yeast-derived biochar, the model compound for
easily degradable biochar stimulated soil microorganisms most
in both soils leading to soil organic carbon losses twice as high
as in the controls, whereas glucose-derived biochar led to
Table 6 intermediate soil organic carbon losses in both soil types.
Summary of analysis of variance (ANOVA) of the principal component analyses of Increasing soil organic carbon losses caused by charcoal input
microbial community composition.
have been observed also in other studies (Wardle et al., 2008).
Parameter Sum of squares F-value Significance The mechanism behind still has to be resolved. However, even if
PC 1 Soil 0.00 0.00 1.000 carbon turnover was increased by the biochar treatments in our
Charcoal 15.94 7.97 <0.001 experiment we want to point out that the total carbon content in
Soil  Charcoal 0.00 0.00 0.741
the soil still was 27% higher in glucose-biochar treatments and
PC 2 Soil 0.00 0.00 1.000 23% in yeast-biochar treatments compared to the controls at the
Charcoal 10.87 13.92 0.001 end of our experiment regardless of the soil type. Calculated
Soil  Charcoal 0.44 0.57 0.581
mean residence times of 4–29 years lead to the conclusion that
1308 S. Steinbeiss et al. / Soil Biology & Biochemistry 41 (2009) 1301–1310

18 18
AG FG
16 AY 16 FY
A F
isotopic shift in ‰ (treatment - inital) 14 14

12 12

10 10

8 8

6 6

4 4

2 2

0 0
fungi gram(+) gram(-) bacteria fungi gram(+) gram(-) bacteria

Fig. 7. Isotopic shift of PLFA biomarkers (treatment after incubation – initial values) for certain microbial groups, i.e. fungi, Gram-negative bacteria, Gram-positive bacteria and
bacteria in general. Error bars reflect the standard deviation in isotopic shift within a group of microorganisms.

biochars produced by hydrothermal pyrolysis would probably Inherited soil microorganisms adapted to the new carbon source
add to the decadal soil carbon pool. The fast decrease in soil and utilized both types of biochar. The biochar type determined,
respiration rates during incubation indicated that microbial which group of microorganisms were involved in the decomposi-
stimulation and thus decomposition processes remarkably tion process. Yeast-derived biochar strongly promoted fungi, while
slowed down after 4 months. Concluding from this observation, glucose-derived biochar primarily was utilized by Gram-negative
we assume that any fertilizer effect of these biochars will be bacteria.
biggest directly after biochar addition to the soils and that the
carbon storage function will gain importance on the longer term. 3) Is the stability of these biochars tuneable by varying the
Mean residence times given here represent results from the condensation grade and chemical composition of the
initial phase of biochar degradation and might increase with biochar?
time.
As has been shown for the total carbon stocks, also the Our results clearly show that the type of biochar, i.e. conden-
reaction of soil microorganisms on biochar addition was driven sation grade and chemical structure, is the main driver for all
by the biochar type and largely independent of the soil type. differences observed between our treatments. All patterns
Yeast-derived biochar strongly increased the proportion of fungi observed for the biochar types were the same in both soils. We thus
in both soils, which consequently turned out as the microbial conclude that the condensation grade and the chemical structure of
group that most utilized this type of biochar. Glucose-derived biochars produced by this method could serve as ‘‘tuning param-
biochar was used as carbon source for the build up of bacterial eter’’ to design biochars that act as fertilizers but simultaneously
biomass. Bacteria in arable soil, probably better adapted to add to the soil carbon pool on a decadal time scale.
carbon limitation events and more complex remaining soil In our current experiment we manipulated the condensation
carbon, respired more biochar carbon than bacteria in forest soil. grade and the nitrogen content of the biochars with promising
Consequently, calculated mean residence times of glucose- results. Several other elements (phosphorus, sulphur, cations)
derived biochar in arable soil were slightly shorter than in forest could be introduced to the biochar structure helping to fill special
soil. Microorganisms in the forest soil seemed to be less nutrient demands of arable lands. Further studies are necessary to
specialized on certain carbon sources. The adaptation of certain design the best possible soil amendments and to investigate the
microbial groups to biochar degradation was less pronounced long-term behavior of these biochars in natural systems.
than in arable soil.
To finally answer the questions that should be solved with our
experiment:
Acknowledgements
1) How stable are biochars produced by this method in different
soil? This investigation was financially supported by the Max Planck
Society within the scope of the EnerChem project house. We
Biochars produced by hydrothermal pyrolysis would add to the thank Maria M. Titirici for her help with SEM measurements at
decadal soil carbon pool. the Max Planck Institute of Colloids and Interfaces in Potsdam-
Golm. Valerian Ciobota and Petra Roesch kindly recorded the
2) How do inherited soil microorganisms react on the addition of Infrared-spectra at the Institute of Physical Chemistry of the
such biochars? Friedrich Schiller University in Jena.
 S. Steinbeiss et al. / Soil Biology & Biochemistry 41 (2009) 1301–1310 1309

Appendix 1

Amounts of identified PLFAs in mg g1 soil dw and d13C values in & in initial soils, all treatments and the controls after incubation.
Standard deviations are given in parentheses. Source specific summaries can be found at the end of each section.
PLFA Source AI (mg g1 dw) AG (mg g1 dw) AY (mg g1 dw) A (mg g1 dw) FI (mg g1 dw) FG (mg g1 dw) FY (mg g1 dw) F (mg g1 dw)
C14:0 br Gram(þ) 1.8 (0.1) 0.4 (0.2) 0.7 (0.2) 2.5 5.1 (0.7) 2.5 (0.4) 2.3 (0.1) 2.1
C14:0 Bacteria 1.1 (0.1) 0.4 (0.1) 0.7 (0.1) 1.5 1.9 (0.2) 1.3 (0.2) 1.5 (0.1) 1.2
C15:1 Gram() 1.4 (0.1) 0.4 (0.2) 0.6 (0.1) 1.9 2.4 (0.2) 0.9 (0.1) 0.9 (0.0) 0.8
C15:0 br Gram(þ) 6.6 (0.6) 2.8 (1.2) 4.3 (0.7) 9.1 10.9 (0.9) 7.7 (1.2) 8.2 (0.4) 6.2
C15:0 br Gram(þ) 5.5 (0.5) 2.3 (0.9) 3.1 (0.5) 7.6 14.8 (1.0) 10.6 (1.7) 10.5 (0.6) 8.5
C15:0 Bacteria 0.6 (0.0) 0.4 (0.1) 0.6 (0.1) 0.8 0.8 (0.1) 0.7 (0.1) 0.7 (0.0) 0.5
C16:0 br Gram(þ) 0.6 (0.1) 0.4 (0.1) 0.4 (0.1) 0.8 0.9 (0.2) 0.9 (0.1) 0.7 (0.0) 0.7
C16:1 Gram() 1.2 (0.1) 0.5 (0.2) 0.6 (0.1) 1.7 1.8 (0.1) 1.0 (0.2) 0.9 (0.1) 0.8
C16:0 br Gram(þ) 3.6 (0.4) 1.8 (0.7) 2.1 (0.5) 5.1 5.4 (0.3) 4.2 (0.8) 4.2 (0.2) 3.1
C16:1 Gram() 2.4 (0.2) 1.1 (0.3) 1.7 (0.3) 3.4 4.2 (0.2) 1.7 (0.3) 2.0 (0.2) 1.5
C16:1 Gram() 10.6 (0.9) 4.6 (1.2) 5.6 (1.1) 14.6 11.8 (0.5) 4.9 (0.7) 5.6 (0.4) 4.8
C16:1 Gram() 6.0 (0.5) 3.1 (0.6) 3.4 (0.7) 8.3 8.5 (0.4) 3.1 (0.4) 3.2 (0.2) 2.8
C16:0 Bacteria 11.4 (1.2) 7.7 (1.5) 15.9 (3.3) 16.1 20.5 (0.9) 15.6 (2.4) 31.1 (2.9) 14.3
C17:0 br Gram(þ) 1.3 (0.1) 0.8 (0.2) 0.8 (0.1) 1.7 2.1 (0.1) 1.1 (0.1) 1.2 (0.1) 0.9
C17:1 Gram() 4.0 (0.4) 2.3 (0.6) 2.8 (0.6) 5.5 5.5 (0.3) 2.7 (0.4) 2.8 (0.2) 2.3
C17:0 br Gram(þ) 5.9 (0.5) 3.9 (0.7) 3.6 (0.9) 8.2 11.2 (0.5) 5.2 (0.9) 4.7 (0.3) 4.2
C17:0 br Gram(þ) 1.3 (0.1) 0.9 (0.1) 0.9 (0.1) 1.7 1.9 (0.1) 0.9 (0.1) 0.9 (0.1) 0.8
C17:0 br Gram(þ) 1.2 (0.1) 0.8 (0.1) 0.7 (0.2) 1.6 1.8 (0.1) 0.9 (0.1) 0.9 (0.0) 0.8
C17:0 br Gram(þ) 2.3 (0.2) 1.7 (0.4) 1.9 (0.3) 3.1 4.2 (0.2) 3.1 (0.5) 3.2 (0.2) 2.6
C17:0 br Gram(þ) 2.5 (0.2) 1.6 (0.3) 1.7 (0.4) 3.6 4.1 (0.2) 2.8 (0.4) 2.9 (0.2) 2.3
C17:1 Gram() 2.0 (0.2) 1.1 (0.3) 0.9 (0.2) 2.8 1.5 (0.1) 0.9 (0.1) 0.8 (0.0) 0.7
C17:1 Gram() 2.8 (0.2) 2.0 (0.3) 2.0 (0.4) 3.8 5.9 (0.2) 3.4 (0.5) 4.1 (0.3) 3.2
C17:0 Bacteria 0.5 (0.1) 0.4 (0.1) 0.5 (0.1) 0.7 0.7 (0.1) 0.7 (0.1) 0.7 (0.1) 0.5
C18:0 br Gram(þ) 2.3 (0.2) 1.8 (0.9) 1.7 (0.5) 3.2 3.0 (0.2) 2.7 (0.5) 2.4 (0.1) 1.9
C18:0 br Gram(þ) 0.5 (0.0) 0.3 (0.1) 0.3 (0.1) 0.6 0.7 (0.0) 0.4 (0.1) 0.4 (0.0) 0.3
C18:0 br Gram(þ) 1.0 (0.1) 0.8 (0.3) 0.7 (0.2) 1.5 1.4 (0.1) 1.5 (0.3) 1.2 (0.1) 1.0
C18:2u6,9 Fungi 3.4 (0.3) 1.6 (0.3) 15.9 (3.3) 4.7 3.5 (0.2) 1.8 (0.3) 27.3 (3.4) 1.5
C18:1u9 Fungi 13.5 (1.4) 7.4 (1.3) 22.0 (4.6) 19.0 25.7 (1.0) 15.5 (2.6) 39.7 (4.1) 13.6
C18:1u11 Gram() 21.2 (2.1) 14.1 (2.2) 16.6 (3.9) 29.3 46.7 (1.6) 26.2 (3.9) 31.1 (2.7) 23.2
C18:1 Gram() 1.8 (0.1) 1.2 (0.1) 1.4 (0.3) 2.8 2.9 (0.3) 1.3 (0.1) 1.7 (0.1) 1.2
C18:0 Bacteria 2.7 (0.3) 2.1 (0.4) 3.3 (0.8) 3.8 4.8 (0.3) 3.7 (0.6) 6.0 (0.5) 3.5
C18:0 cyc Gram() 1.7 (0.2) 1.3 (0.2) 1.6 (0.3) 2.3 3.2 (0.1) 3.0 (0.4) 3.7 (0.3) 2.8
C19:0 br Gram(þ) 2.6 (0.3) 1.8 (0.6) 2.3 (0.7) 3.6 4.8 (0.3) 4.0 (0.8) 4.6 (0.3) 2.9
C19:0 br Gram(þ) 0.7 (0.0) 0.6 (0.1) 0.5 (0.1) 1.1 1.3 (0.1) 1.0 (0.1) 1.0 (0.1) 0.7
C18:0 cyc Gram() 5.5 (0.6) 4.5 (0.5) 4.2 (1.0) 7.7 22.9 (0.9) 12.2 (2.0) 12.3 (0.9) 10.7
C20:1u9 Gram() 1.2 (0.1) 0.7 (0.2) 1.1 (0.2) 1.6 2.2 (0.2) 1.1 (0.2) 1.7 (0.2) 1.0
C20:0 Bacteria 0.7 (0.0) 0.4 (0.0) 0.6 (0.1) 0.9 1.0 (0.1) 0.8 (0.1) 1.1 (0.1) 0.8

Sum Gram() 61.8 36.9 42.5 85.7 119.5 62.4 70.8 55.8
Gram(þ) 39.7 22.7 25.7 55.0 73.5 49.5 48.3 39.0
Fungi 16.9 9.0 37.9 23.7 29.2 17.3 67.0 15.1
Bacteria 16.5 11.4 21.6 23.8 29.7 22.8 41.1 20.8

d13C (&) d13C (&) d13C (&) d13C (&) d13C (&) d13C (&) d13C (&) d13C (&)
C14:0 br Gram(þ) 29.4 (1.4) 26.6 (1.3) 29.4 (0.4) 28.5 (1.5) 30.6 (3.4) 28.1 (0.3)
C14:0 Bacteria 28.8 (0.3) 22.6 (0.5) 24.6 (1.3) 23.2 (0.9) 23.1 (0.9) 25.6 (1.2) 24.3 (0.0)
C15:1 Gram() 26.6 (2.1) 23.0 (0.6) 24.3 (0.5) 24.2 (0.3) 24.2 (0.2)
C15:0 br Gram(þ) 25.0 (0.6) 25.0 (1.1) 21.7 (0.2) 23.6 (1.4) 24.7 (0.3) 23.1 (1.0) 23.1 (1.2) 23.3 (0.5)
C15:0 br Gram(þ) 21.2 (0.2) 19.1 (1.2) 19.0 (0.3) 21.5 (0.7) 20.4 (0.2) 18.8 (0.2) 16.8 (0.6) 19.3 (0.3)
C15:0 Bacteria 27.8 (1.6) 23.0 (1.2) 25.9 (1.3) 25.7 (0.5) 26.0 (0.5)
C16:0 br Gram(þ) 22.5 (3.3) 22.8 (0.4) 25.3 (1.0) 28.0 (1.9) 24.3 (1.8) 26.4 (0.9)
C16:1 Gram() 27.1 (1.5) 21.4 (1.8) 24.5 (1.4) 26.3 (0.5)
C16:0 br Gram(þ) 25.2 (0.3) 23.7 (1.4) 22.5 (0.4) 25.9 (1.3) 23.8 (0.9) 25.5 (0.1)
C16:1 Gram() 21.3 (0.3) 19.0 (0.9) 17.1 (0.3) 20.8 (0.2) 23.4 (0.9) 20.3 (0.4) 19.5 (0.8) 20.2 (0.1)
C16:1 Gram() 26.1 (0.5) 20.5 (1.0) 21.6 (1.7) 24.9 (0.5) 25.9 (0.5) 22.4 (0.7) 21.3 (0.6) 25.1 (0.3)
C16:1 Gram() 20.7 (0.1) 18.8 (0.3) 18.7 (0.3) 21.3 (0.4) 21.5 (0.3) 18.6 (1.9) 17.4 (0.9) 20.2 (0.5)
C16:0 Bacteria 24.8 (1.4) 20.8 (0.8) 17.8 (1.0) 23.9 (0.5) 23.8 (0.1) 21.0 (1.1) 18.4 (0.2) 23.2 (0.2)
C17:0 br Gram(þ) 19.0 (1.1) 9.7 (0.8)
C17:1 Gram() 27.1 (1.0) 21.2 (0.6) 24.8 (1.5) 24.6 (0.4) 22.9 (1.6) 24.2 (1.7) 22.1 (0.5)
C17:0 br Gram(þ) 23.9 (0.3) 22.0 (0.2) 21.8 (0.9) 23.0 (0.5) 22.6 (0.4) 21.4 (1.1) 21.1 (1.1) 22.4 (0.6)
C17:0 br Gram(þ) 18.9 (0.7) 15.4 (0.7) 20.2 (0.0) 17.6 (1.7) 19.5 (1.3) 18.1 (0.1) 20.0 (1.3) 17.9 (1.3)
C17:0 br Gram(þ) 22.3 (0.3) 20.3 (0.6) 21.1 (1.1) 22.6 (0.2) 20.6 (1.4) 21.8 (1.3) 21.8 (0.4)
C17:0 br Gram(þ) 22.2 (0.3) 22.3 (0.6) 18.7 (0.4) 20.6 (0.4) 20.4 (0.2) 19.1 (1.1) 19.5 (0.7) 19.4 (0.5)
C17:1 Gram() 23.5 (0.6) 15.9 (1.3) 20.7 (0.6) 22.4 (0.7) 23.7 (0.3) 17.5 (1.7) 19.0 (0.4) 22.2 (0.5)
C17:0 Bacteria 24.4 (0.9) 20.6 (0.6) 22.0 (1.6) 16.6 (0.8) 19.4 (0.6) 21.0 (0.5)
C18:0 br Gram(þ) 25.5 (0.1) 24.2 (0.3) 24.0 (0.7) 26.1 (1.4) 24.3 (0.2) 22.3 (1.6) 22.7 (1.2) 23.9 (0.6)
C18:0 br Gram(þ) 21.6 (1.0) 24.0 (0.3) 20.5 (0.6)
C18:0 br Gram(þ) 27.3 (1.1) 23.9 (1.1) 26.6 (1.6) 22.9 (1.7) 17.0 (2.0) 23.6 (2.5) 20.7 (1.7)
C18:2u6,9 Fungi 31.6 (1.1) 31.0 (0.6) 15.4 (1.2) 29.7 (0.7) 28.0 (0.5) 19.6 (1.3) 18.6 (0.2) 23.2 (1.0)
C18:1u9 Fungi 27.1 (0.7) 24.6 (1.0) 13.8 (0.5) 27.2 (1.6) 23.3 (0.3) 22.2 (0.3) 13.8 (0.3) 22.7 (0.5)
C18:1u11 Gram() 24.8 (0.5) 16.7 (1.2) 17.6 (0.4) 24.7 (1.0) 24.6 (0.2) 19.5 (0.5) 18.2 (0.4) 23.3 (0.2)
(continued on next page)
1310 S. Steinbeiss et al. / Soil Biology & Biochemistry 41 (2009) 1301–1310

Appendix 1 (continued )

PLFA Source AI (mg g1 dw) AG (mg g1 dw) AY (mg g1 dw) A (mg g1 dw) FI (mg g1 dw) FG (mg g1 dw) FY (mg g1 dw) F (mg g1 dw)
C18:1 Gram() 14.9 (0.5) 10.1 (0.8) 13.0 (2.0) 15.2 (1.3) 14.5 (1.1) 8.2 (2.5) 5.5 (1.3) 10.8 (1.0)
C18:0 Bacteria 20.9 (0.7) 18.5 (0.3) 17.5 (0.3) 21.4 (1.2) 21.3 (0.5) 18.9 (0.5) 14.9 (0.3) 20.9 (0.4)
C18:0 cyc Gram() 21.1 (0.9) 16.6 (0.4) 19.4 (0.4) 20.5 (0.6) 23.7 (0.5) 16.9 (0.3) 15.3 (0.4) 21.9 (0.6)
C19:0 br Gram(þ) 29.1 (0.8) 23.3 (1.0) 25.8 (0.8) 22.1 (0.9) 21.7 (1.4) 18.7 (0.2) 21.7 (0.9)
C18:0 cyc Gram() 28.3 (0.5) 25.4 (1.0) 25.6 (1.2) 28.5 (0.8) 27.9 (0.4) 26.4 (0.4) 25.3 (0.5) 27.5 (0.3)
C20:0 Bacteria 21.6 (1.9) 23.6 (1.0) 25.4 (0.1) 20.9 (0.6) 21.5 (1.0)

Average Gram() 23.8 (4.0) 17.9 (4.4) 19.9 (3.3) 22.9 (3.4) 23.4 (3.5) 19.1 (5.1) 19.3 (6.2) 22.2 (4.4)
Gram(þ) 24.2 (3.1) 21.4 (3.3) 22.0 (2.4) 23.4 (2.9) 22.8 (3.2) 20.6 (4.3) 21.8 (3.8) 22.0 (3.5)
Fungi 29.4 (3.2) 27.8 (4.5) 14.6 (1.1) 28.5 (1.8) 25.7 (3.3) 20.9 (1.8) 16.2 (3.4) 23.0 (0.4)
Bacteria 25.3 (3.1) 19.7 (1.6) 20.5 (2.4) 24.0 (1.9) 23.3 (1.5) 21.0 (3.4) 19.8 (3.9) 22.8 (2.1)

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DOI 10.1007/s11104-007-9391-5
MARSCHNER REVIEW
Mycorrhizal responses to biochar in soil – concepts
and mechanisms
Daniel D. Warnock & Johannes Lehmann &
Thomas W. Kuyper & Matthias C. Rillig
Received: 19 April 2007 / Accepted: 9 August 2007 / Published online: 19 September 2007
# Springer Science + Business Media B.V. 2007
Abstract Experiments suggest that biomass-derived
black carbon (biochar) affects microbial populations
and soil biogeochemistry. Both biochar and mycor-
rhizal associations, ubiquitous symbioses in terrestrial
ecosystems, are potentially important in various
ecosystem services provided by soils, contributing to
sustainable plant production, ecosystem restoration,
and soil carbon sequestration and hence mitigation of
global climate change. As both biochar and mycor-
rhizal associations are subject to management, under-
standing and exploiting interactions between them
could be advantageous. Here we focus on biochar
Responsible Editor: Hans Lambers.
D. D. Warnock : M. C. Rillig
Microbial Ecology Program,
Division of Biological Sciences, University of Montana,
Missoula, MT 59812, USA
J. Lehmann
Department of Crop and Soil Sciences, Cornell University,
Ithaca, NY 14853, USA
T. W. Kuyper
Department of Soil Quality, Wageningen University,
P.O. Box 47, 6700 AA Wageningen, The Netherlands
M. C. Rillig (*)
Institut für Biologie, Freie Universität Berlin,
Altensteinstr. 6, 14195 Berlin, Germany
e-mail: rillig@zedat.fu-berlin.de
effects on mycorrhizal associations. After reviewing
the experimental evidence for such effects, we critically
examine hypotheses pertaining to four mechanisms by
which biochar could influence mycorrhizal abundance
and/or functioning. These mechanisms are (in decreas-
ing order of currently available evidence supporting
them): (a) alteration of soil physico-chemical proper-
ties; (b) indirect effects on mycorrhizae through effects
on other soil microbes; (c) plant–fungus signaling
interference and detoxification of allelochemicals on
biochar; and (d) provision of refugia from fungal
grazers. We provide a roadmap for research aimed at
testing these mechanistic hypotheses.
Keywords Biochar . Arbuscular mycorrhiza .
Ectomycorrhiza . Carbon storage . Restoration .
Terra preta
Introduction
Pioneering studies, conducted primarily in Japan,
where biochar application to soil has a long tradition
(Ishii and Kadoya 1994), provided evidence that
biochar can have positive effects on the abundance
of mycorrhizal fungi (Table 1). Soil micro-organisms,
especially arbuscular mycorrhizal fungi (AMF), in
addition to ectomycorrhizal fungi (ECM) and ericoid
mycorrhizal fungi (ERM), have well-recognized roles
in terrestrial ecosystems (Zhu and Miller 2003; Rillig
 10

Table 1 Effects of biochar (BC) or activated carbon/charcoal (AC) additions on mycorrhizal fungi, separated by mycorrhizal type (arbuscular mycorrhizal fungi (AMF)
ectomycorrhizal fungi (ECM), and ericoid mycorrhizal fungi (ERM), and listed in order of decreasing effect size of the mycorrhizal response variable(s)

Experimental designa Amouont ACb or BCb present Type(s) of BCc or ACc applied Response variablesd Mycorrhiza responsee Possible functions for Source
ECM, ERM or AMFf

AMF experiments
BC effects on AMF RC of BC: 800 g/m3 in 2, 4.8 m3 H: RH RC +610% ND Ishii and
Citrus iyo in an abandoned pits Kadoya
orchard (F) (1994)
Effects of three BC types on BC: 2.0% BW H: RH Citrus Juice RC +540% RH +88% Enhanced overall Ishii and
AMF (Glomus fasciculatum) Sediment (C.J.) C.J. +75% W.S. plant P nutrition Kadoya
in river sand (G) Woody: Western (1994)
Spruce Bark (W.S.)
BC Effects on AMF in soy bean BC: 1,500 g m−2 ND RC +300% ND Saito (1990)
fields (F)
BC (ground vs un-ground) BC: 33% BV H: RH RC Ground: +100% ND Ezawa et al.
effects on AMF infectivity (F) Un-ground: −20% (2002)
BC effects on AMF (Glomus sp.) BC : 10% and 30% BV Woody: Coconut Shell RC 10% BC: +50% 30% Enhanced plant Matsubara
and Fusarium oxysporum RC of BC: +69% pathogen resistance et al. (2002)
Asparagus officinalis roots. (G)
BC effects on infectivity of BC: Applied at a rate Woody: Acacia RC +42% ND Yamato et al.
indigenous AMF (G) of 10 l m−2 mangium bark (2006)
BC effects on AMF RC of non BC: Applied at rates of 0, Woody: Eucalyptus RC Non N-fixing: 30 g, ND Rondon et al.
N-fixing, and N-fixing 30, 60 and 90 g BC kg−1 soil deglupta logs 60 g: −38% 90 g: −20% (2007)
Phaseolus vulgaris) roots. (G) N-fixing: 30 g, 60 g:
NS; 90 g: +16%
BC Effects on AMF RC, and BC: 89.8% BV of ND RC SD in 100 ml−1 RC −21% SD: −5% ND Gaur and
Spore density (S.D.) by Glomus growth substrate infectious IP: −38% Adholeya
intraradices grown in culture propagules (IP) (2000)
with Zea mays (G) in 100 ml−1
ECM experiments
Quantified ECM RC in different BC: 2% BV ND RC, no. ECM +2,900% ND Harvey et al.
soil fractions of a Montana root tips 100 cm3 (1976)
forest soil (F) soil fraction −1
Effect of AC on timing of AC: 2% BW ND RC Onset of RC +624% Onset Colonization by P. Herrmann et
mycorrhizal colonization of mycorrhiza accelerated by croceum increased al. (2004)
Quercus robur seedlings formation 4 weeks drought resistance
by Piloderma croceum.(G) measured in weeks in Q. robur
AC effects on ability of ECM AC: 0.3% BV ND ECM presence or +200% ND Vaario et al.
(Pisolithus tinctorus) to absence of host (1999)
colonize Abies firma seedlings infection
Plant Soil (2007) 300:9–20

grown in culture (G)

Table 1 (continued)

Experimental designa Amouont ACb or BCb present Type(s) of BCc or ACc applied Response variablesd Mycorrhiza responsee Possible functions for Source
ECM, ERM or AMFf

Effectiveness of RH BC/forest BC: 300 cm3 BC H: RH Presence or absence +80% ND Mori and
top soil mix as ECM inoculum mixed with 1 l soil. of host infection Marjenah
source for Shorea smithiana BC/soil mix placed in by ECM fungi (1994)
trees grown in degraded forest potting hole 25-cm deep×
soil. (F) 25-cm diameter
Effects of AC slurry on dissolved AC: Applied to soil as ND RC −38% in type ND Wellstedt
phenol concentration and Picea slurry, (250 g AC 3 l−1 B fungi et al. (2002)
mariana seedling growth (G) water) microcosm
Plant Soil (2007) 300:9–20

surface area=1,890 cm2

ERM experiments
Effect of AC only, or AC and AC: Added to solid agar Darcco G60, RC +95% AC ND Duclos and
carbon source (0.5 g l−1 glucose medium at 1 g l−1 Fisher +128% AC + Fortin (1983)
or pectin) additions on ERM Glucose, or AC +
RC of Vaccinium angustifolium Pectin
a
G, Greenhouse; F, Field
b
BV, By volume; BW, By weight
c
AC is produced via one of the following activation procedures, CO2, steam, or chemical (e.g. phosphoric acid). All three processes remove remaining organic compounds and
nutrients from previously pyrolyzed biomass while greatly increasing carbonyl content, yielding a porous material with an extremely high surface area and a very high sorptive
capacity. Because the AC activation process begins with charred biomass, until further evidence is provided to the contrary, it is assumed that BC and AC will both act similarly as
adsorbents, in the soil environment. However, AC will likely have a much greater surface area than BC (Pan and van Staden 1998). H, Herbaceous biochar; RH, Rice husk biochar
d
RC, root colonization; SD, spore density
e
NS, non significant difference; effect size for response variables was calculated as ((Xtreatment −Xcontrol)/Xcontrol)×100
f
ND, Not determined
11
12
2004; Read et al. 2004; Rillig and Mummey 2006).
Mycorrhizal fungi are frequently included in manage-
ment, since they are widely used as soil inoculum
additives (Schwartz et al. 2006). With both biochar
additions and mycorrhizal abundance subject to
management practices, there clearly are opportunities
for exploiting a potential synergism that could
positively affect soil quality.
While data on biochar effects on mycorrhiza are
accumulating, there are several important gaps in our
knowledge on these interactions. The most important
gap concerns the mechanisms by which biochar might
affect the abundance and functioning of mycorrhizal
fungi. Therefore, the goals of this paper are to first
evaluate the evidence of biochar effects on mycorrhi-
zal associations thus far, and then to propose
mechanisms for these biochar effects on mycorrhizae
(primarily using examples of arbuscular mycorrhiza
and ectomycorrhiza). In doing so, we also point out
future research priorities (Fig. 1). To clarify the
nomenclature used throughout this discussion we first
provide a brief overview of biochar properties.
Fig. 1 Schematic representation of bio-char and its direct and
indirect effects on mycorrhizal fungi abundance/functioning,
emphasizing the hierarchical nature of effects. The numbers
included in figure body correspond to mechanisms discussed in
text: (1) effects on soil physio-chemical properties; (2) effects
Plant Soil (2007) 300:9–20
Biochar definition and properties
Biochar is a term reserved for the plant biomass-
derived materials contained within the black carbon
(BC) continuum. This definition includes chars and
charcoal, and excludes fossil fuel products or geo-
genic carbon (Lehmann et al. 2006). Materials
forming the BC continuum are produced by partially
combusting (charring) carbonaceous source materials,
e.g. plant tissues (Schmidt and Noack 2000; Preston
and Schmidt 2006; Knicker 2007), and have both
natural as well as anthropogenic sources. Restricting
the oxygen supply during combustion can prevent
complete combustion (e.g., carbon volatilization and
ash production) of the source materials. When plant
tissues are used as raw materials for biochar produc-
tion, heat produced during combustion volatilizes a
significant portion of the hydrogen and oxygen, along
with some of the carbon contained within the plant’s
tissues (Antal and Gronli 2003; Preston and Schmidt
2006). The remaining carbonaceous materials contain
many poly-aromatic (cyclic) hydrocarbons, some of
through influences on other soil microbes; (3) interactions with
plant–fungus signaling; and (4) provision of refugia from
fungal grazers. Solid arrows indicate direct facilitative effects;
dashed arrows indicate indirect facilitative effects
Plant Soil (2007) 300:9–20
which may contain functional groups with oxygen or
hydrogen (Schmidt and Noack 2000; Preston and
Schmidt 2006). Depending on the temperatures
reached during combustion and the species identity
of the source material, a biochar’s chemical and
physical properties may vary (Keech et al. 2005;
Gundale and DeLuca 2006). For example, coniferous
biochars generated at lower temperatures, e.g. 350°C,
can contain larger amounts of available nutrients,
while having a smaller sorptive capacity for cations
than biochars generated at higher temperatures, e.g.
800°C (Gundale and DeLuca 2006). Furthermore,
plant species with many large diameter cells in their
stem tissues can lead to greater quantities of macro-
pores in biochar particles. Larger numbers of macro-
pores can for example enhance the ability of biochar
to adsorb larger molecules such as phenolic com-
pounds (Keech et al. 2005).
Because of its macromolecular structure dominated
by aromatic C, biochar is more recalcitrant to
microbial decomposition than uncharred organic
matter (Baldock and Smernik, 2002). Biochar is
believed to have long mean residence times in soil,
ranging from 1,000 to 10,000 years, with 5,000 years
being a common estimate (Skjemstad et al. 1998;
Swift 2001; Krull et al. 2003). However, its recalci-
trance and physical nature represent significant
obstacles to the quantification of long-term stability
(Lehmann 2007).
Evidence for biochar effects on mycorrhizal fungi
From the experiments summarized in Table 1, it
appears that the addition of biochar materials to soil
often results in significant responses by both plants
and mycorrhizal fungi.
Tryon (1948), Matsubara et al. (2002), DeLuca et
al. (2006), and Gundale and DeLuca (2006) demon-
strated that biochar additions can change soil nutrient
availability by affecting soil physico-chemical prop-
erties. Increases in soil nutrient availability may result
in enhanced host plant performance and elevated
tissue nutrient concentrations in addition to higher
colonization rates of the host plant roots by AMF
(Ishii and Kadoya 1994). Lastly, experiments by
Matsubara et al. (2002) suggested that biochar can
also increase the ability of AMF to assist their host in
resisting infection by plant pathogens.
13
In three of the six ECM studies and the single
ERM study represented in Table 1, experiments
demonstrated the effects of adding biochar in growth
media on both the ability of the ECM and ERM fungi
to colonize the host plant seedlings, and the overall
effects on seedling growth. Additionally, the experi-
ment conducted by Herrmann et al. (2004) showed
that activated carbon (AC), which may in many cases
have similar properties as biochar, affected the timing
of host plant colonization by ECMF, which occurred
4 weeks earlier in the AC treatment than in the
control. The other ECM related experiments evaluated
the effects of biochar presence on host tree coloniza-
tion rates (Harvey et al. 1976; Mori and Marjenah
1994). In these two cases, the presence of biochar
corresponded with significant increases in plant root
colonization by ECM. Observations made by Harvey
et al. (1978, 1979) also support these results.
In contrast to those experiments in Table 1 showing
positive effects of biochar or AC additions on
abundance of mycorrhizal fungi, a few studies
observed negative effects. In these cases, it appears
that the negative effects of the biochar or AC additions
on AMF were largely due to nutrient effects. For
example, Gaur and Adholeya (2000) found that the
biochar media limited the amount of P taken up by
host plants, compared to rates from plants grown in
river sand or clay-brick granules, suggesting that P
was less available. Additionally, Wallstedt et al. (2002)
reported decreases in both bio-available organic
carbon and nitrogen in their ectomycorrhizal system.
An important consideration pertains to the study
design of the experiments reported in Table 1. The
first issue deals with the soils used in the experiments,
e.g. river sand or OM-rich field soil; the other issue
concerns the materials added to these soils as controls,
e.g. organic matter vs biochar. Are soil biota,
including mycorrhizal fungi, responding to an exper-
imental addition of biochar simply because carbon is
being added or are they responding to biochar’s
unique properties? In at least two cases where data
from field soils were presented, it appears that
mycorrhizal fungi responded more positively to
biochar additions than to additions of other types of
organic material added as control (Harvey et al. 1976;
Ishii and Kadoya 1994). The experiment by Matsubara
et al. (2002) showed that a fresh organic amendment
had fairly similar effects as biochar in increasing AMF-
mediated host plant resistance against Fusarium and
14
that the asparagus plants reached similar mycorrhizal
colonization levels with both additions. But the 9-week
gap between inoculation with AMF and with Fusarium
makes this aspect of the experiment somewhat difficult
to evaluate. However, it is still possible that these
positive responses shown by mycorrhizal fungi are
determined in part by the amount of carbon in the
material being added to the soil, with the expectation
that the biochar is more carbon-rich than the organic
matter. We may not be able to answer this question
satisfactorily until experiments control for C amend-
ment effects in the biochar treatment(s) and/or take into
account the relative addition of C to soils.
Work on terra preta de índio (TP) soil, the fertile
Amazonian Dark Earths, has served as a major
inspiration for the use of biochar as a promising soil
additive promoting crop growth and carbon storage
(Glaser et al. 2002; Glaser and Woods 2004;
Lehmann et al. 2006; Glaser 2007). However, no
published data are available on the impact of TP soils
on mycorrhizal functioning. For that reason, the
studies discussed above refer to short-term experi-
ments and not to the historical, pre-Columbian
Amazonian soils. TP soils are not only much richer
in biochar than the surrounding soils, but also in non-
pyrogenic carbon and nutrients, especially phospho-
rus and calcium; therefore it is likely that TP effects
on mycorrhizal functioning could be beyond those of
biochar addition alone.
Mechanisms
At least four mechanisms could explain how biochar
can lead to altered total abundance and/or activity of
mycorrhizal fungi in soils and plant roots: (1) Biochar
additions to soil result in altered levels of nutrient
availability and/or other alterations in soil physico-
chemical parameters that have effects on both plants
and mycorrhizal fungi. (2) Additions of biochar to
soils result in alterations with effects that are
beneficial or detrimental to other soil microbes, for
instance mycorrhization helper bacteria (MHB) or
phosphate solubilizing bacteria (PBS). (3) Biochar in
soils alters plant–mycorrhizal fungi signaling process-
es or detoxifies allelochemicals leading to altered root
colonization by mycorrhizal fungi. (4) Biochar serves
as a refuge from hyphal grazers. Since a primary goal
Plant Soil (2007) 300:9–20
of this discussion is identifying mechanisms explain-
ing the effects of biochar on mycorrhizae, with the
intention of guiding attempts for developing methods
to exploit them as soil management tools, and because
many of the biochar effects included in Table 1 appear
positive, we primarily present arguments explaining why
biochar generally appears beneficial to mycorrhizae.
However, as discussed previously, biochar appli-
cations do not always benefit mycorrhizal fungi (see
Table 1). In these situations, one could argue that
biochar, via any of our proposed mechanisms, reduces
formation of mycorrhiza, e.g. by decreasing nutrient
availability or creating unfavourable nutrient ratios in
soils (Wallstedt et al. 2002). This negative effect
could be especially prominent in cases where the
biochar has a very high C/N ratio and a portion of the
biochar is decomposable, leading to N-immobiliza-
tion. Under such conditions, biochar could also
negatively affect plant growth, e.g. as seen in Gaur
and Adholeya (2000). Given the above possibilities
for negative responses by both plants and mycorrhizal
fungi to biochar amendments, and plants to mycor-
rhizal fungi (Johnson 1993), it cannot be assumed that
biochar amendments will always result in a net
benefit to plant productivity even though few such
cases have been reported so far.
A conceptual overview of the mechanisms and
hypothesized pathways discussed in the following
sections is provided in Fig. 1, emphasizing the
hierarchical nature of contributing factors. In the
following discussion it should be kept in mind that
(a) mechanisms are not mutually exclusive but likely
several contribute to the outcome, perhaps even with
opposite effects; (b) there is little information avail-
able on which mechanism is likely the most important
in any given environmental situation; and finally that
(c) many mechanisms are hypothetical with most
support for mechanism 1 at this time (we are
presenting mechanisms below in decreasing amount
of evidence). This figure therefore also serves as a
roadmap for future research.
Mechanism 1: Biochar changes soil nutrient
availability
Modifications of nutrient availability would clearly be
a mechanism of primary importance for mycorrhizal
fungal abundance. For example, nutrient additions
might alleviate growth limitations of the fungi
Plant Soil (2007) 300:9–20
themselves in nutrient-poor soils (Treseder and Allen
2002). Additionally, altering the balance of nutrients
can exert strong control over fungal root colonization,
as for example known for shifts in soil N/P ratios for
AMF (Miller et al. 2002).
Biochar addition can result in elevated quantities of
bio-available nutrients such as N, P and metal ions, in
the affected soils (Tryon 1948; Lehmann et al. 2003;
Gundale and DeLuca 2006; DeLuca et al. 2006), but
has also been shown to lead to decreases particularly
of N availability (Lehmann et al. 2003). These
changes in soil nutrient availabilities, may be
explained by some of the following observations.
Additions of biochar to soil alters important soil
chemical and physical (see below) properties such as
pH (has caused both increases and decreases), and
typically increase soil cation exchange capacity
(CEC), and can lead to greater water holding capacity
(WHC), while generally decreasing bulk density
(Tryon 1948). Increases in soil pH towards neutral
values (Lucas and Davis 1961), in addition to
increased CEC (Glaser et al. 2002), may result in
increases in bio-available P and base cations in
biochar influenced soils. Additionally, Lehmann et
al. (2003), Topoliantz et al. (2005), Gundale and
DeLuca (2006) and Yamato et al. (2006) showed that
biochar itself contained small amounts of nutrients
that would be available to both soil biota (including
mycorrhizal fungi) and plant roots. Lastly, DeLuca et
al. (2006) showed that biochar from forest wildfire
stimulated gross and net nitrification rates, most likely
mediated by biochar adsorbing inhibitory phenols.
This mechanism is likely specific to soils with
ectomycorrhizal trees and/or ericaceous shrubs with
an abundance of phenolic compounds, whereas in
agricultural soils biochar may in the short term reduce
ammonification and nitrification by a reduction either
in N availability due to immobilization during initial
decomposition of the N-poor biochar (Lehmann et al.
2006) or by a reduction in C cycling.
Some of the experiments conducted to evaluate
the effects of biochar upon mycorrhizae (Table 1)
lend support to mechanism 1. These experiments
show that additions of biochar materials generally
result in the alteration of soil physico-chemical
properties that may lead to increases in soil nutrient
availability (measurements taken from both soil
samples and plant tissues) and/or increases in root
colonization by mycorrhizal fungi (Ishii and Kadoya
15
1994; Matsubara et al. 2002; Yamato et al. 2006). In a
greenhouse experiment by Matsubara et al. (2002),
the soil pH of treatments receiving biochar increased
from 5.4 to 6.2 (10% biochar by volume) and 6.3
(30% biochar by volume). According to Lucas and
Davis (1961), these pH values fall within the pH
range (5.5 to 7.0) where plant nutrients are near their
maximum availability in agricultural soils. Many of
these alterations in soil characteristics probably occur
at a micro-scale (Gundale and DeLuca 2006), and thus
may only affect hyphae that are in the immediate
vicinity of biochar particles.
Mechanism 2: Biochar alters the activity of other
micro-organisms that have effects on mycorrhizae
Mycorrhization Helper Bacteria (MHB; Garbaye 1994)
are capable, under specific conditions, of secreting
metabolites, e.g. flavonoids (AMF) and furans (ECM),
that facilitate the growth of fungal hyphae and the
subsequent colonization of plant roots by ECM
(Founoune et al. 2002; Duponnois and Plenchette
2003; Aspray et al. 2006; Riedlinger et al. 2006) and
AM fungi (Duponnois and Plenchette 2003; Hildebrandt
et al. 2002, 2006). Hildebrandt et al. (2002, 2006) have
demonstrated that certain compounds (including raffi-
nose and other unidentified metabolites) produced by
strains of Paenibacillus can directly enhance the
growth of AMF extraradical mycelium. Additionally,
Kothamasi et al. (2006) demonstrated that other species
of bacteria, such as Pseudomonas aeruginosa, can
solubilize important plant nutrients, especially phos-
phate, making them part of a group of bacteria called
phosphate solubilizing bacteria (PSB). These mineral-
ized nutrients are then accessible to mycorrhizal fungi
and eventually to the host plant. Furthermore, Xie et al.
(1995) and Cohn et al. (1998) state that Rhizobium sp.
and Bradyrhizobium sp. can produce compounds that
induce flavonoid production in nearby plants (legumes)
that may ultimately increase root colonization of plant
roots by AM fungi.
Biochar may serve as a source of reduced carbon
compounds (either the biochar particle itself, or organic
molecules adsorbed to the particle’s matrix), and/or
nutrients, and as a refuge (see mechanism 4) for any
biochar colonizing soil bacteria, including MHB and
PSBs (Pietikäinen et al. 2000; Samonin and Elikova
2004). Increased populations of PSB and/or MHB
might then indirectly benefit mycorrhizal fungi (Fig. 1).
16
Mechanism 3: Biochar alters the signaling dynamics
between plants and mycorrhizal fungi or detoxifies
allelochemicals
The rhizosphere is a zone of intense signaling
between microbes, including mycorrhizal fungi, and
plant roots (Bais et al. 2004; Harrison 2005; Bais et
al. 2006; Paszkowski 2006). For example, experiments
conducted using both field soils and in vitro cultures
show that compounds (e.g. CO2, flavonoids, sesquiter-
penes and strigolactones) secreted by plant roots lead to
both increased colonization of plant roots by AMF
(Bécard and Piché 1989; Nair et al. 1991; Xie et al.
1995) and increased spore germination and AMF hyphal
branching (Gianinazzi-Pearson et al. 1989; Akiyama et
al. 2005). Additions of biochar could alter the exchange
of signals in several ways, as shown in Fig. 1; however,
we emphasize that most of the pertinent evidence stems
from sterile in vitro culture studies with uncertain
relevance to conditions in the soil.
Angelini et al. (2003) demonstrated that some
flavonoid signaling compounds could be either
inhibitory or stimulatory to specific groups of soil
biota as a function of pH. As discussed under
mechanism 1, biochar additions usually increase soil
pH. Hence, it is possible that these pH changes alone
can lead to stimulatory effects, causing increases in
fungal abundance.
Sorptive properties of biochar (e.g. for hydropho-
bic substances), particularly higher temperature (e.g.,
800°C) biochar, could also cause signaling interfer-
ence in the rhizosphere: biochar could serve as signal
reservoirs or as a sink, both for signaling compounds
and for inhibitory compounds (allelochemicals). Re-
cently, Akiyama et al. (2005) demonstrated that AC
was capable of adsorbing AMF signaling (strigolac-
tones) compounds from a hydroponic solution that
were subsequently desorbable with acetone. Once
desorbed, these compounds retained their activity and
stimulate hyphal branching and growth of Gigaspora
margarita. Biochar particles could adsorb signal
molecules not immediately intercepted by AMF
hyphae or spores, or consumed by other soil biota.
Later on, these stored signal molecules could be
desorbed by soil water reaching the biochar particles.
After being re-dissolved into soil water, they would
again be available to stimulate mycorrhizal coloniza-
tion of plant roots. By functioning in this manner,
biochar particles would be serving as secondary
Plant Soil (2007) 300:9–20
sources of signal molecules, acting concomitantly
with MHB and plant roots.
However, biochar’s capacity to adsorb signaling
compounds and add as a sink could also decrease the
ability of mycorrhizal fungi to colonize plant roots. If
biochar permanently rather than temporarily removes
signal molecules from soils, this signal sorption
activity results in a net decrease in the number of
signal molecules reaching mycorrhizal hyphae and
spores. As a result, hyphal growth and spore germina-
tion, and ultimately fungal abundance, could actually
decrease because of biochar activity.
In addition to chemical signals, biochar may also
adsorb compounds toxic to mycorrhizal fungi. For
example, Wallstedt et al. (2002) showed that the
addition of an AC slurry to an experimental soil
resulted in a decreased amount of water-soluble
phenols. Herrmann et al. (2004) and Vaario et al.
(1999) related their results of stimulated ECM fungus
colonization of roots in the presence of AC to toxin
sorption.
Mechanism 4: Biochar serves as a refuge
for colonizing fungi and bacteria
This mechanism is purely physical in nature, and
therefore could function in a similar fashion for ECM,
ERM, AMF and bacteria. Hyphae and bacteria that
colonize biochar particles (or other porous materials)
may be protected from soil predators (Saito 1990;
Pietikäinen et al. 2000; Ezawa et al. 2002), which
includes mites, collembola and larger (>16 μm in
diameter) protozoans and nematodes. The docu-
mented physical parameters of the biochar particles
themselves make this mechanism plausible. The
average sizes of soil bacteria and fungal hyphae range
from 1 to 4 μm and 2 to 64 μm, respectively, with
many fungal hypha being smaller than 16 μm in
diameter (Swift et al. 1979). Additionally, the average
body-size of a soil protist is between 8 to 100 μm,
while the average body size of soil micro-arthropods
ranges from 100 μm to 2 mm (Swift et al. 1979). In
contrast, the pore diameters in a biochar particle can
often be smaller than 16 μm in diameter (Kawamoto
et al. 2005; Glaser 2007; Hockaday et al. 2007).
Based on the differences in the body sizes across
these different organisms, it is clearly possible that
many of the pores within a biochar particle are large
enough to accommodate soil microorganisms, includ-
Plant Soil (2007) 300:9–20
ing most bacteria and many fungi, to the exclusion of
their larger predators. Thus, the biochar would be
acting as a refuge for MHB, PSB and mycorrhizal
fungi. Supporting evidence for this hypothesis comes
from Saito (1990), Gaur and Adholeya (2000) and
Ezawa et al. (2002) who all showed that AMF readily
colonize porous materials and were capable of heavily
colonizing biochar particles in the soil. Lastly,
Pietikäinen et al. (2000) and Samonin and Elikova
(2004) showed that bacteria readily colonized biochar
particles; these may include MHB and/or PSB.
An important factor controlling pore size distribution
is the charring temperature with higher temperatures
yielding finer pores. Another major factor in determining
the degree to which biochar may serve as a refuge is the
anatomical structure of the biological tissues pyrolyzed
to yield the biochar. Considering the effects that cell
diameter alone can have on the sorptive capability of a
given biochar material (Keech et al. 2005; Gundale and
DeLuca 2006), it stands to reason that the cell types
contained within the original plant tissues (e.g.,
tracheids, vessel elements or sieve cells) determine the
pore sizes of the biochar. Not only the charring
conditions and source material, but also subsequent
interactions of biochar with soil can change porosity
and pore sizes. For example, adsorption of organic
matter to biochar surfaces can decrease porosity by
blocking pores (Kwon and Pignatello 2005).
While it seems clear that mycorrhizal fungi can use
biochar as a habitat, the quantitative importance to the
extraradical mycelium is not evident. This will highly
depend on the biochar properties and the biochar
addition rates. Nevertheless, the finer parts of the
mycelium, generally the absorptive hyphae, are more
vulnerable to fungal grazers (Klironomos and Kendrick
1996), and it is primarily these architectural elements
that could be effectively protected within biochar
particles. It would depend, then, on the extent to which
these ‘protected’ fine hyphae make a substantial
contribution towards nutrient uptake compared to the
relatively ‘unprotected’ hyphae in the mineral and
organic soil, whether this hypothesized mechanism is
quantitatively important.
Conclusions and research recommendations
Experimental results (Table 1) point to exciting
possibilities regarding biochar and its possible syner-
17
gy with arbuscular, ericoid, and ectomycorrhizal
symbioses. We have synthesized available data into
several potential mechanisms of biochar effects on
mycorrhizae (Fig. 1). This should serve as a spring-
board for testing the occurrence and relative impor-
tance of these factors/mechanisms in the soil. Based
on this discussion we derive the following research
recommendations:
(a) Methods reporting. In many cases it is helpful to
know as much detail about the experimental
biochar application as possible. This should in-
clude: source material, production temperature,
application rate, application method, and what
material was used in the control application to
account for C addition effects (and the amounts of
available nutrients for both). This would facilitate
comparisons among studies and help distinguish
among the different mechanistic pathways; fre-
quently these pieces of information are incomplete.
(b) Management implications. None of the studies to
date have examined the management context of
biochar application on AMF, and this would also be
an important research need, since application
practices could have overriding effects on soil biota.
(c) Fungal communities. Studies to date have focused
on quantifying potential responses in fungal abun-
dance measures, primarily root colonization and
spore numbers (see Table 1). However, mycorrhi-
zal fungi occur as species assemblages in ecosys-
tems and in roots of individual plants (Johnson et
al. 1992; Husband et al. 2002; Vandenkoornhuyse
et al. 2003; Mummey et al. 2005). The species
composition of a mycorrhizal fungal assemblage
can be important to mycorrhizal functioning (e.g.,
van der Heijden et al. 1998). Data on this
important aspect of the response of mycorrhizal
fungi to biochar are not yet available, but represent
an important priority for future studies. Here, we
limited our discussion to mechanisms affecting
abundance; however, many of the arguments
presented could also be applied to explain poten-
tial shifts in mycorrhizal fungal species composi-
tion, because fungal life history strategies and
responsiveness to changing soil environments vary
between fungal taxa (e.g., Hart and Reader 2002;
Escudero and Mendoza 2005; Drew et al. 2006).
(d) Negative effects. There is a potential for negative
effects on mycorrhizal fungi, as discussed above;
18
it is therefore clearly also a research priority to
define the environmental circumstances (e.g., soil
nutrient content, plants species) and biochar
parameters (e.g., quality and application rate) that
lead to such effects. It is possible that negative or
neutral effects have been under-reported.
Increasing atmospheric concentrations of carbon
dioxide have prompted the search for avenues of
long-term sequestration of carbon, particularly in the
soil (Lal 2004; Schiermeier 2006). Work on terra
preta de índio soil has inspired the use of biochar as a
promising soil additive promoting carbon storage
(Day et al. 2005; Lehmann et al. 2006; Marris 2006;
Glaser 2007). Biochar can add value to non-harvested
agricultural products (Major et al. 2005; Topoliantz et
al. 2005), and can promote plant growth (Lehmann et
al. 2003; Oguntunde et al. 2004). Lehmann et al.
(2006) estimated that a total of 9.5 billion tons of
carbon could potentially be stored in soils by the year
2100 using a wide variety of biochar application
programs. Once equipped with a better understanding
of this potential synergism and the mechanisms that
drive it, we could utilize biochar/mycorrhizae inter-
actions for sequestration of carbon in soils to
contribute to climate change mitigation. This interac-
tion could also be harnessed for the restoration of
disturbed ecosystems, the reclamation of sites con-
taminated by industrial pollution and mine wastes,
increasing fertilizer use efficiencies (with all associ-
ated economic and environmental benefits) and the
development of methods for attaining increased crop
yields from sustainable agricultural activities.
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Impact of Biological Growths
(personal communication D. Day 2005)

- AM Fungi lifespan of ~3 weeks

- Glomalin lifespans averages 24 years, carbon in aggregates can be shorter but still ~18 years. (300x the
life of the fungi

- Total Soil Organic Carbon ~1500Gt

- With 7% of SOC =carbon from glomalin (CFG), = ~105Gt (Nichols)

With ~32% of SOC =carbon in aggregates resulting from glomalin, = ~480Gt (Nichols)

- An amount directly tied to AM Fungi and glomalin growth = ~31Gt of the carbon from glomalin and
carbon resulting from glomalin (CRFG), classed as in the annual pool of flux.
- CFG/CRFG probably does not rise linearly with glomalin growth but probably does increase as 50% of the
total increase in AM fungi rise.

- There are 93.5M km2 of biologically active land (UNEP) then ~334ton km-1 yr-1 of CFG/CRFG are in
annual flux.

- No till studies in the U.S. have found that soil carbon increases (mid level results) at 163 ton km-1 yr-1.

- Assuming charcoal addition can provide a 200% increase in AM fungi growth (Low end of Ishii's findings)

- And at 50%, doubles CFG production to 334 ton km-1 yr-1

- If 33% of the land were dedicated to energy crops and and converted to hydrogen or other fuel (7% yield
@ ~2.5x107j kg-1), replacing coal (5.8x106j kg-1), and would replace 219 ton C km-1 of fossil C with C
from bio-energy production.

- Then 2.2 x10E6 km2 (2.0% of the earth's biologically active land) would need to be treated once every 3
yrs with a ton per hector with low temp char (or equivalent nutient enhanced charcaol) and using notill
farming to create a sink to offset the 1.7Gt buildup.

- A 400% AM fungi increase, then we would need to treat 1.5x10E6 km2 (1.6% of the earth biologically
active land or 2.8 times the size of France). It is possible these amounts are not required annually since
there is a significant carry over benefit from year to year. Ogawa (Japan) recommends only once every
three years.
Environ Sci Pollut Res (2009) 16:1–9
DOI 10.1007/s11356-008-0041-0
AREA 1.2 • CHEMICALS IN SOILS/SEDIMENTS • RESEARCH ARTICLE
Reduction of nitrogen loss and Cu and Zn mobility
during sludge composting with bamboo charcoal amendment
Li Hua & Weixiang Wu & Yuxue Liu &
Murray B. McBride & Yingxu Chen
Received: 28 December 2007 / Accepted: 10 August 2008 / Published online: 27 August 2008
# Springer-Verlag 2008
Abstract
Background, aim, and scope Composting is an effective
treatment process to realize sludge land application.
However, nitrogen loss could result in the reduction of
nutrient value of the compost products and the stabilization
effect of composting on heavy metal concentration and
mobility in sludge has been shown to be very limited.
Materials and methods Laboratory-scale experiments were
carried out to investigate the effects of bamboo charcoal
(BC) on nitrogen conservation and mobility of Cu and Zn
during sludge composting.
Results The result indicated that the incorporation of BC
into the sludge composting material could significantly
reduce nitrogen loss. With 9% BC amendment, total
nitrogen loss at the end of composting decreased 64.1%
compared with no BC amendment (control treatment).
Mobility of Cu and Zn in the sludge may also have been
lessened, based on the decline in diethylenetriaminepenta-
acetic acid-extractable Cu and Zn contents of composted
sludge by 44.4% and 19.3%, respectively, compared to
metal extractability in the original material.
Discussion Ammonia adsorption capability of BC might be
the main reason for the retention of nitrogen in sludge
composting materials. Decrease of extractable Cu2+ and
Responsible editor: Hailong Wang
L. Hua : W. Wu (*) : Y. Liu : Y. Chen
College of Environmental and Resource Science,
Zhejiang University,
Hangzhou 310029, China
e-mail: weixiang@zju.edu.cn
M. B. McBride
Department of Crop and Soil Science, Cornell University,
Ithaca, NY 14853, USA
Zn2+ in the composting material mainly resulted from the
adsorption of both metals by BC.
Conclusions Incorporation of BC into composting material
could significantly lessen the total nitrogen loss during sludge
composting. Mobility of heavy metals in the sludge compost-
ing material could also be reduced by the addition of BC.
Recommendations and perspectives Bamboo charcoal
could be an effective amendment for nitrogen conservation
and heavy metal stabilization in sludge composts. Further
research into the effect of BC-amended sludge compost
material on soil properties, bioavailability of heavy metals,
and nutrient turnover in soil needs to be carried out prior to
the application of BC-sludge compost in agriculture.
Keywords Amendment . Bamboo charcoal .
Mobility of Cu and Zn . N conservation .
Sludge composting
Abbreviations
BC Bamboo charcoal
FBC Fresh bamboo charcoal
CBC Composted bamboo charcoal
TOM Total organic matter
TOC Total organic carbon
1 Background, aim, and scope
At least 15 million tons (78% water content) of municipal
sewage sludge are produced annually in China. Due to the
absence of sound treatment methods, large amounts of
sludge have been stockpiled at temporary suburban storage
sites. Many people regard land application as a cost-
effective method for the disposal and beneficial use of
2 Environ Sci Pollut Res (2009) 16:1–9
sludge (Sigua et al. 2005; Su et al. 2007). However, the
presence of potentially toxic chemicals such as heavy
metals in sludge could pose risks to the environment and
human health (Illera et al. 2000; Hseu 2006). Composting is
regarded as an effective treatment process to suppress
pathogenic organisms and stabilize the organic material in
sewage sludge (Fang and Wong 1999; Hernández et al.
2006). However, the stabilization effect of composting on
heavy metal mobility in sludge has been shown to be very
limited (Amir et al. 2005; Liu et al. 2007). On the other
hand, nutrient loss is an unavoidable problem during the
composting of organic waste. This is especially the case for
nitrogen, the most essential fertilizer nutrient for crop
production in many situations (Cooperband and Middleton
1996; Rao Bhamidimarri and Pandey 1996). According to
previous research, total nitrogen (TN) loss during the course
of composting organic waste ranged from 16% to 76%
(Barrington et al. 2002; Raviv et al. 2002). Substantial loss
of nitrogen results in the reduction of nutrient value of
compost products. Among all practicable nitrogen conserva-
tion measures, an amendment was thought to be the most
efficient method to reduce nitrogen loss during composting.
For example, alum, peat, and zeolite amendments have been
used to reduce ammonia volatilization during composting
(Bernal et al. 1993; Kithome et al. 1999). Lime and coal fly
ash have also been applied as amendments to reduce the
mobility of heavy metals in composting mixtures (Fang and
Wong 1999; Qiao and Ho 1997). Nevertheless, low
efficiency, potentially negative influence on soil pH, and
high cost are factors that have prevented these amendments
from being adopted broadly in composting technology.
Bamboo charcoal (BC) is one kind of manufactured
biocharcoal, a plentiful residual byproduct of the bamboo
processing industry. BC has a highly microporous physical
structure. The porosity is about five times greater and the
absorption efficiency ten times higher than that of wood
charcoal (Zhang 2001). Bamboo charcoal may be an ideal
amendment for nutrient conservation and heavy metal
stabilization due to its excellent adsorption capability.
Recent research found that biocharcoals could act as soil
Table 1 Characteristics of sludge, rapeseed marc, and bamboo charcoal
Sludge Rapeseed marc
Total nitrogen (%) 2.31±0.15 Total nitrogen (%)
Organic matter (%) 34.32±2.26 Organic matter (%)
Total phosphorus (%) 1.5±0.10 Total phosphorus (%)
pH 7.41±0.15 pH
Water content (%) 78.72±2.2 Water content (%)
Total Zn (mg kg−1) 1,530±56 Total Zn (mg kg−1)
Total Cu (mg kg−1) 270±15 Total Cu (mg kg−1)
C/N 16±1 C/N
ND No detection
fertilizers or conditioners to increase crop yield and plant
growth by supplying and retaining nutrients (Glaser et al.
2000; Major et al. 2005; Steiner et al. 2007). However,
there has been no research to date on the potential
application of bamboo charcoal in nutrient conservation
and heavy metal stabilization during sludge composting.
The main objectives of this study, therefore, are to
investigate the potential capability of BC to stabilize heavy
metals and retain nitrogen during sludge composting and to
clarify the possible mechanisms related to these functions.
We assume that the results of this study will provide
practical information to guide the exploitation of a novel
compost amendment in agricultural applications.
2 Materials and methods
The sewage sludge was collected from Fuyang wastewater
treatment plant located in a suburb of Hangzhou city,
China. It treats more than 100,000 m3 of wastewater
annually and produces about 90 tons day−1 of sludge
(80% water content). The bamboo charcoal particles used in
this study were purchased from the Yaoshi Charcoal-
Production Company located in Hangzhou. Rapeseed marc
was purchased from a local farm, which was used as a
bulking agent in sludge composting due to its higher bulk
density and lower moisture content. Characteristics of
sludge, bamboo charcoal, and rapeseed marc are listed in
Table 1. Cu and Zn were chosen as sludge metals for study,
because they are quantitatively prevalent metals in sludges
and are known to have higher bioavailability than less
soluble metals such as Pb and Cr according to previous
studies (Oliveira et al. 2007; Dai et al. 2007).
2.1 Composting and sampling
Composting was carried out in a tank with dimensions of
1.0×0.4×1.0 m and a volume of approximately 0.4 m3.
There were six different treatments. The stock material
consisted of 180-kg sewage sludge and 20-kg rapeseed
Bamboo charcoal
1.01±0.05 Pyrolysis temperature (°C) 600
73.62±4.22 Density (g cm−3) 0.75±0.04
0.45±0.02 Specific surface (m2 g−1) 330±24
8.23±3.55 pH 7.32±0.1
8.30±4.24 Water content (%) 10±1
ND Total Zn (mg kg−1) 0.45±0.03
ND Total Cu (mg kg−1) ND
129±5 C/N 356±10
Environ Sci Pollut Res (2009) 16:1–9
marc with 0%, 1%, 3%, 5%, 7%, 9% (w/w) of BC
incorporated into the stock material. These BC treatments
are labeled as control, 1% BC, 3% BC, 5% BC, 7% BC,
and 9% BC. Aeration was ensured by forced ventilation
during the first 3 weeks of composting and by turning the
pile over manually during the remaining incubation time.
Temperature of the composting pile was recorded daily
using temperature sensors. Each sampling was carried out
in triplicate. All together, six times of samplings were
performed.
2.2 Chemical analyses
Total nitrogen was measured by the Kjeldahl digestion
method (Hernández et al. 2006). Total organic matter was
determined by potassium dichromate (K2Cr2O7) and sulfu-
ric acid (H2SO4) oxidation (Dai et al. 2007). Pile mass was
based on the total mass of all the composting material
including sludge, bamboo charcoal, and rapeseed marc at
the initial and the final composting stages. Based on the N
concentration and the total pile mass before and after
composting, the actual N loss could be obtained. The actual
N amount was calculated by multiplying the N concentra-
tion and the total pile mass of the composting material. The
actual N loss was the difference between the initial N
amount and the final N amount in the composting materials.
Boehm titration (Boehm 1994) was used to quantify
various surface functional groups on the fresh BC (FBC)
particle and composted BC (CBC; separated from mixed
final composting materials) particle. A solar Thermo MkII-
6 atomic absorption spectrometer with deuterium back-
ground corrector was used to determine the contents of
heavy metal in compost samples. To determine the total
concentration of heavy metals, 0.5 g of sample was
digested in a 20-mL mixture of nitric, hydrochloric, and
hydrofluoric acid (in the ratio 1:1:2; Scancara et al. 2000).
Diethylenetriaminepentaacetic acid (DTPA)-extractable
metal contents were obtained by mechanically shaking the
compost samples at a 1:5 solid to extractant ratio (w/v) for
2 h with 0.005 M DTPA+0.1 M triethanolamine solution
buffered at pH 7.3 (Page et al. 1982).
2.3 Cu2+, Zn2+, and NH4+ adsorption by fresh BC
and composted BC
Twenty milliliters of Cu2+, Zn2+ solutions with the concen-
trations of 200–2,000 mg L−1, or NH4+ solutions with
concentrations of 2–10 mg L–1 were added to 0.2 g of FBC/
CBC in order to find out if the change of surface
characteristic would result in the change of retaining
capability of bamboo charcoal to these ions. Copper
sulphate pentahydrate (CuSO4·5H2O), zinc sulphate hepta-
hydrate (ZnSO4·7H2O), and ammonium chloride (NH4Cl)
3
were used as stock chemicals in the BC adsorption
experiments and the concentrations were dependent on the
actual concentration in sludge of these ions. The solutions
with BC particle were then kept in a thermostatic shaker at
250 rpm and 25°C for 48 h to achieve equilibrium.
Afterwards, each sample was filtered to separate the FBC/
CBC particles from solution. These particles were analyzed
to determine amounts of Cu Zn and ammonium adsorbed
and used for desorption tests. For the desorption experi-
ment, the residual FBC or CBC samples collected from the
adsorption experiment were extracted by 0.05 M DTPA
solution to measure the strength of Cu2+ and Zn2+ retention
on FBC/CBC. The detection process was the same as
described in the adsorption experiment. The heavy metal
retention capability was expressed as the percent of the total
Cu and Zn in BC that was retained after extracting by
DTPA. The FBC/CBC particles which had already reached
ammonium adsorption equilibrium were extracted with
30 mL of 0.01 M CaCl2 (Nishantha Fernando et al. 2005)
using the same conditions as described for adsorption. The
ammonium retention capability was expressed by the same
approach described for metals. In the adsorption–desorption
test, triplicated sampling were implemented for the analysis
of the bamboo charcoal in initial composting material and
final composting material.
2.4 Data analysis
All data were expressed as means and standard deviations
compared statistically by Tukey’s t test at the 5% level with
SPSS11.5 (SPSS for windows, version 11.5, USA). Any
differences between values with p>0.05 are not considered
to be statistically significant.
3 Results
3.1 Chemical changes of material during composting
Temporal trends of the temperature within the composting
materials were quite similar regardless of the level of BC
amendment (Fig. 1). The compost temperature rose to 55°C
within 5 days and remained above 55°C for around 7 days.
Afterwards, the compost temperature gradually declined and
reached ambient temperature in 30 days. Finally, the compost
went into maturation stage until composting was completed.
Total organic matter of the composting materials
experienced a relatively fast decline at the early stage of
composting and then decreased steadily over time. There
were no significant differences in total organic matter of the
composting materials among different treatments except in
the early stage. By the end of the experiment, about 30% of
the initial total organic matter had mineralized (Fig. 2). In
4 Environ Sci Pollut Res (2009) 16:1–9
Fig. 1 Change of temperature during the course of composting
contrast, trends in the change of TN in composting
materials over the composting process varied with treat-
ment. TN concentrations in the composting material with
lower BC amendment (<3%) decreased during the thermo-
philic phase and then increased afterwards. When the BC
amendment rose to more than 5%, TN concentrations in the
composting material increased over the entire composting
process (Fig. 3). A first-order kinetic equation was used to
calculate the potentially mineralizable C (C0):

increased quickly in the first 4 weeks and became stable
Cm ¼ C0 1
e
kt
where Cm (mg C/kg) is the organic C mineralized at any
specific time, t (days), and k is the first-order rate constant.
Similarly, the increase of N with time could be expressed
as a first-order kinetics equation. Overall, the TN loss
(based on mass balance) in the composting material
Fig. 2 Change of total organic matter during the course of
composting
Fig. 3 Change of total nitrogen concentration during the course of
composting
decreased with an increasing amount of BC amendment.
The highest TN loss (674 g) was found in composting
material with no BC amendment, whereas TN loss in the
treatment with 9% BC amendment was only 240 g, a
difference of more than 64% (Table 2).
Boehm titration is one of the most widely used methods
to quantify acidic groups in charcoal. Based on this method,
the amount of total acid groups of BC underwent a
significant change during the course of composting, which
after 28 days of composting. Among all acidic groups, the
amount of carboxylic groups increased most. As shown in
Table 3, the amount of carboxylic groups of BC increased
up to 0.69 mmol g−1 at the end of composting, which was
about 2.4 times higher than that of the original value
(0.29 mmol g−1). However, the amounts of phenolic and
lactonic groups on BC surface increased only about 50%
compared with those of the original values.
3.2 Changes of DTPA-extractable Cu and Zn contents
in the composting material
Sewage sludge contains various kinds of heavy metals.
However, Cu and Zn are quantitatively primary heavy
metals in sludge of China, as well as in sludge of other
countries (Udom and Mbagwu 2004; Dai et al. 2007).
Relative percentages of total Cu and Zn in the composting
material that were DTPA-extractable during the composting
process are shown in Figs. 4 and 5. In general, DTPA-
extractable contents of Cu and Zn in the composting
material decreased with composting time. The decrease of
DTPA-extractable Cu in the composting materials was
greater than that of DTPA-extractable Zn content during the
composting process. By the end of composting, DTPA-
extractable Cu and Zn contents in the composting material
Environ Sci Pollut Res (2009) 16:1–9 5

Table 2 Mass balances and losses of nitrogen during composting of sludge

Treatment Initial material Finished material N loss (g) N loss rate (%)

Pile mass (kg) Content (g kg−1) N mass (g) Pile mass (kg) Content N mass (g)
(g kg−1)

Control 200 8.3 1,660.0 105. 9.30 983.6 676.4 40.7

1% BC 202 8.21 1,658.4 106 9.63 1,022.76 635.6 38.3
3% BC 206 8.07 1,662.6 109 10.36 1,127.2 535.4 32.2
5% BC 210 7.94 1,667.4 113 10.83 1,219.5 447.9 26.7
7% BC 214 7.82 1,673.5 117 11.32 1,327.8 345.7 20.4
9% BC 218 7.63 1,663.3 120 11.87 1,423.2 240.1 14.6

without BC amendment had decreased 29.2% and 12.0%,
respectively, compared with the initial value before com-
posting began. Furthermore, significant effects of BC
amendment on the reduction of DTPA-extractable Cu and
Zn in composting material were observed during compost-
ing, with the effects differing considerably depending on
the BC amendment level. For DTPA-extractable Cu, a
significant difference between the BC amendments and the
non-BC control was detected when the addition of BC was
higher than 3%. For DTPA-extractable Zn, a difference
between the BC amendments and the non-BC control was
found only when the BC amendment was larger than 5%.
Compared with the original value before composting, the
DTPA-extractable Cu and Zn in the treatment with 9% BC
amendment decreased 44.4% and 19.3%, respectively, over
the course of composting. Overall, in contrast to the non-
BC control, DTPA-extractable Cu and Zn contents in
composting materials could be further reduced by 27.5%
and 8.2%, respectively, with 9% BC amendment.
3.3 Cu2+, Zn2+, and NH4+ adsorption by BC before
and after composting
As shown in Table 4, the amount of Cu adsorbed onto BC
was much higher than that of Zn, which was consistent with
the fact that DTPA-extractable content of Cu decreased more
than that of Zn. For example, in solutions with 2,000 mg L−1
of Cu2+ and Zn2+, the amounts of Cu2+ adsorbed on FBC
was 36.0 mmol g−1, whereas Zn2+ adsorbed on FBC was
only 14.2 mmol g−1. Results in Table 4 also indicate that the
retention of Cu2+ and Zn2+ by FBC and CBC decreased as
initial adsorption of these two metals increased. The Cu2+
retention against DTPA extraction on FBC and CBC
decreased from 88.0% to 74.0% and 88.2% to 75.0%,
respectively. Retention of Zn2+ on FBC and CBC decreased
from 69.8% to 53.5% and 70.0% to 52.1%, respectively.
However, according to the result of Tukey’s t test, no
difference was observed between the retention capability of
FBC and CBC for Cu2+ and Zn2+. According to Nishantha
Fernando et al. (2005), the retaining of NH4+ against Ca2+
exchange can express the retaining capacity of absorbent to
NH4+; therefore, CaCl2 was adopted as extracting solvent in
desorption test to determinate the retaining rate of BC to
NH4+. In this research, the retention of NH4+ against Ca2+
exchange on FBC decreased with increasing adsorption on
BC (Table 5), a pattern of behavior similar to that of Cu2+
and Zn2+. However, the retention of NH4+ on CBC did not
decrease significantly with an increase of adsorption. A
significantly greater retention of NH4+ was observed by
CBC compared to FBC, which indicated that composting has
a significantly positive effect on the adsorption capability of
BC for NH4+.

Table 3 Amount of surface functional groups determined by Boehm titration (p<0.05)

Sample Carboxylic groups Phenolic and lactonic groups (mmol g–1) Total acidic functional groups (mmol g–1)
(mmol g–1)

Original BC 0.29e±0.03 0.84c±0.03 1.17d±0.05

BC after 7 days composting 0.36de±0.04 0.88c±0.07 1.26d±0.07
BC after 14 days composting 0.46cd±0.05 1.02b±0.06 1.48c±0.04
BC after 21 days composting 0.55bc±0.04 1.10b±0.04 1.66b±0.05
BC after 28 days composting 0.62ab±0.03 1.24a±0.05 1.86a±0.03
BC after 35 days composting 0.67a±0.05 1.26a±0.07 1.94a±0.03
BC after 42 days composting 0.69a±0.06 1.27a±0.08 1.95a±0.02

Amount values of surface functional groups followed by different small letters within the same vertical column are different significantly at 5%
level
6 Environ Sci Pollut Res (2009) 16:1–9
Fig. 4 Change of DTPA-extractable Cu content during the course of
composting
4 Discussion
Bamboo charcoal amendment has little effect on the
microbial activity and organic matter mineralization during
sludge composting, based on the measured temporal
changes in temperature and organic matter content. How-
ever, TN loss in the composting material varied depending
on the amount of BC amendment. Overall, nitrogen loss
decreased as the amount of BC incorporated into the
composting materials increased. It is generally known that
surface area and pore size distribution, as well as
heteroatoms and compounds affecting surface properties,
affect the adsorption capacity (Tennant and Mazyck 2007).
As shown in Table 1, the specific surface and bulk density
of BC were 330 m2 g−1 and 0.75 g cm−3, indicating that BC
has a substantial adsorption capability. It is probably this
high adsorption capability resulted in the considerable
reduction of TN loss with the BC amendment during and
after sludge composting. Specifically, ammonia adsorption
Fig. 5 Change of DTPA-extractable Zn content during the course of
composting
capability of BC might be the main reason for the retention
of nitrogen in sludge composting materials, as 47–77% of
nitrogen loss has been reported as resulting from ammonia
volatilization during composting (Martins 1992). The
reductions of TN content in composting materials with
lower amount of BC amendment during the thermophilic
period were probably attributable to an insufficient concen-
tration of BC particles to adsorb ammonia emitted rapidly
by the degradation of proteins and amino acids (Pagans et
al. 2006). However, ammonia binding to complex organic
materials involved in the humification process has a role in
preventing ammonia release and volatilization at the
maturation stage (Paredes et al. 2002; Baddi et al. 2004).
As a result, the TN concentration recovered to higher
values in the composting materials as weight loss due to the
mineralization of organic matter continued in the matura-
tion phase.
According to adsorption theory, the adsorbate occupies
sites where it is more exchangeable or labile first by a fast
reaction and then migrates to sites where it is less
exchangeable over time. Therefore, after the sites with
nonexchangeable adsorbate are filled, additional adsorbate
will remain exchangeable, suggesting that adsorbate would
be released more easily as greater loading occurs on the
adsorbent. In this study, the NH4+ retention rate of FBC
decreased as the amount of adsorption increased, indicating
the lower stability of adsorption at higher levels. The
saturation of strong binding sites at high adsorption
amounts could explain the phenomenon. However, results
for the CBC desorption test showed that there were no
significant differences in NH4+ retention rate regardless of
amount of NH4+ adsorbed, which indicated that composting
could improve the nitrogen conservation capacity of BC.
As the results of Boehm titration have shown, the amount
of total acidic groups, especially carboxylic groups, which
would retain NH4+ through ionic bonding, increased
substantially during the course of composting. As is well
known, ionic bonding is more energetic than simple
physical adsorption. This could explain the high retention
rate of NH4+ on CBC, even for treatments with higher
added NH4+. Therefore, we conclude that the bio-oxidation
of BC during sludge composting could increase NH4+
conservation, with this increase mainly attributed to the
increased acid groups on the BC surface. Nishantha
Fernando et al. (2005) stated that the carboxylic groups
may deprotonate and therefore form complexes with NH4+,
and phenolic constituents could react with NH4+ to form
stable complexes. Some authors had also reported the
nitrogen retention properties of other biocharcoals. For
example, Oya and Iu (2002) and Iyobe et al. (2004) found
black charcoal in soil could efficiently adsorb ammonia and
act as a buffer for ammonia in soil, suggesting that it has
the potential to decrease ammonia volatilization from
Environ Sci Pollut Res (2009) 16:1–9 7

Table 4 Results of adsorption–desorption tests of Cu and Zn on fresh bamboo charcoal and composted bamboo charcoal (p<0.05)

Concentration Cu Zn
of solution
for adsorption FBC CBC FBC CBC
test (mg L−1)
Sorption Retention rate Sorption Retention rate Sorption Retention rate Sorption Retention rate
value (%) value (%) value (%) value (%)
(mmol g−1) (mmol g−1) (mmol g−1) (mmol g−1)

200 6.14e±0.15 87.99a±2.32A 6.01e±0.29 88.18a±3.25A 4.94e±0.41 69.78a±2.11A 4.33d±0.29 70.0a±2.15A

400 15.01d±0.25 87.25a±2.54A 14.23d±0.28 86.58a±3.11A 6.40d±0.15 68.76a±2.35A 6.49c±0.23 68.85a±2.55A
800 27.49c±0.84 81.57b±2.23A 25.82c±0.76 82.68b±2.15A 8.97c±0.81 63.64b±1.46A 8.81b±0.6 63.36b±2.02A
1,200 31.54b±1.68 80.81b±2.10A 29.80b±0.60 79.95b±1.20A 10.63b±0.31 64.54b±1.96A 9.67b±0.17 63.17b±2.50A
1,600 34.48a±1.37 74.58c±2.47A 33.89a±0.28 75.18c±2.16A 13.23a±1.15 59.96c±2.38A 12.30a±0.21 58.87c±2.29A
2,000 35.99a±2.04 74.02c±3.18A 34.61a±0.56 74.96c±3.12A 14.22a±0.73 53.53d±2.24A 13.25a±0.86 52.10d±2.10A

Sorption values followed by different small letters within the same vertical column are different significantly at 5% level. Retention rate followed
by different capital letter of BC and CBC transversely are different significantly at 5% level

agricultural fields. However, there have been no studies
prior to this research about the effect of biocharcoal and
especially BC on nitrogen conservation during the course of
composting.
DTPA-extractable content of heavy metals can estimate
the mobility of heavy metals in most cases (Hseu 2006;
Fang and Wong 1999). In comparison with the original
values before composting, DTPA-extractable contents of
Cu2+ and Zn2+ in treatment with 9% BC decreased by
44.4% and 19.3%, respectively, over the course of
composting. In contrast to the non-BC control, DTPA-
extractable contents of Cu2+ and Zn2+ in composting
material could be further decreased by 27.5% and 8.2%,
respectively, with the 9% BC amendment. This additional
decrease of extractable Cu2+ and Zn2+ in the composting
material mainly resulted from the adsorption of both metals
by BC. The findings of Hiller and Brümmer (1997) that
charcoal particles tended to accumulate heavy metals
support this hypothesis. In addition, the results of this
study indicated that BC played a more efficient role in
stabilization of Cu2+ than Zn2+. This stabilization discrep-
ancy might result from the different adsorption stability of
BC to Cu2+ and Zn2+, which may correlate with the greater
tendency for covalent bonding (higher electronegativity) of
Cu. In addition, the results of adsorption–desorption tests
showed that the retention of Cu2+ and Zn2+ on FBC and
CBC decreased with increasing adsorbate loading. Accord-
ing to the adsorption theory mentioned above, this decrease
could be explained by saturation of the strong binding sites
on the absorbent. Moreover, there was no significant
difference in Cu2+ and Zn2+ retention by FBC compared
to CBC, thus suggesting that BC oxidation during
composting had no evident effect on the adsorption of
Cu2+ and Zn2+ by BC.
5 Conclusions
Incorporation of BC into composting material could
significantly lessen the total nitrogen loss during sludge
composting. TN loss decreased with an increasing amount
of BC amendment. In comparison to the non-BC control,
the amendment of 9% BC reduced TN loss from the sludge
composting material by 64.1% after composting. This
reduction of TN loss mainly resulted from the high
adsorption capacity of BC and the bio-oxidation of BC

Table 5 Results of adsorption–desorption tests of ammonium on fresh bamboo charcoal and composted bamboo charcoal (p<0.05)

Concentration of solution for FBC CBC

adsorption test (mg L−1)
Sorption value (mmol g−1) Retention rate (%) Sorption value (mmol g−1) Retention rate (%)

2 0.53d±0.02 84.4a±3.08A 0.56c±0.02 88.2a±2.30A

4 0.81c±0.04 83.8a±3.14B 0.92b±0.05 88.8a±2.82A
6 1.05b±0.04 76.6b±2.26B 1.15a±0.05 87.7a±3.29A
8 1.17a±0.03 75.1b±2.85B 1.22a±0.03 87.7a±3.45A
10 1.20a±0.02 70.9c±2.56B 1.23a±0.05 88.1a±2.44A

Sorption values followed by different small letters within the same vertical column are different significantly at 5% level. Retention rate followed
by different capital letters of BC and CBC transversely are different significantly at 5% level
8 Environ Sci Pollut Res (2009) 16:1–9
particles during composting, which significantly increased
the amount of surface acid groups, especially carboxylic
groups.
Mobility of heavy metals in the sludge composting could
also be reduced by the addition of BC. However, the
stabilization effect of BC was different for Cu2+ and Zn2+.
DTPA-extractable contents of Cu2+ and Zn2+ in sludge
composting material with 9% BC amendment dropped
27.5% and 8.2%, respectively, at the end of composting as
compared with that of the non-BC control. There was no
significant difference in the ability of FBC compared to
CBC to retain either Cu2+ or Zn2+, indicating that
composting had little effect on the adsorption capacity of
BC for these heavy metals.
6 Recommendations and perspectives
According to the results of the study, it is concluded that
BC could be an effective amendment for nitrogen conser-
vation and heavy metal stabilization in sludge composts.
However, further research into the effect of BC-amended
sludge compost material on soil properties, bioavailability
of heavy metals, and nutrient turnover in soil needs to be
carried out prior to the application of BC-sludge compost in
agriculture.
Acknowledgements This study was partially supported by China
National Natural Science Fund (40432004), Project of Science and
Technology Department of Zhejiang Province (2006C13066 and
2007C03002), and Program for New Century Excellent Talents in
University.
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WEH, Zech W (2007) Long term effects of manure, charcoal
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 BIOCHAR VOLATILE MATTER CONTENT EFFECTS ON PLANT
GROWTH AND NITROGEN TRANSFORMATIONS IN A TROPICAL
SOIL

Jonathan L. Deenik, A.T. McClellan and G. Uehara

Department of Tropical Plant and Soil Sciences, University of Hawaii, Honolulu, HI

ABSTRACT
Biochars made from modern pyrolysis methods have attracted widespread attention as
potential soil amendments with agronomic value. A series of greenhouse experiments
and laboratory incubations were conducted to assess the effects of biochar volatile
matter (VM) content on plant growth, nitrogen (N) transformations, and microbial
activities in an acid tropical soil. High VM biochar inhibited plant growth and reduced
N uptake with and without the addition of fertilizers. Low VM charcoal supplemented
with fertilizers improved plant growth compared with the fertilizer alone. The
laboratory experiments showed that high VM biochar increased soil respiration and
immobilized considerable quantities of inorganic N. This research shows that biochar
with high VM content may not be a suitable soil amendment in the short-term.

INTRODUCTION
The use of biochar as a soil amendment is modeled on the C-rich anthropogenic soils known
as “Terra Preta do Indio” (Indian black earth) found in Amazonia and associated with habitation
sites of pre-contact Amerindian populations dating as far back as 7,000 cal yr BP (Glaser, 2007).
The defining characteristic of Terra Preta soils is the presence of large quantities of charcoal in
the soil organic matter to depths of 1 m or greater (Glaser et al., 2000; Sombroek et al., 1993).
These soils are remarkable because they have remained fertile and enriched in soil C compared
with adjacent forest soils despite centuries of cultivation.
Recent efforts to replicate the “Terra Preta” phenomenon using biochars created from
modern pyrolysis techniques show that charcoal additions can have an ameliorating effect on
highly weathered, infertile tropical soils by increasing CEC and plant nutrient supply, reducing
soil acidity and aluminum toxicity, and improving fertilizer efficiency due to reduced nutrient
leaching (Glaser at al., 2002; Lehmann et al., 2003). Plant growth response to charcoal amended
soils has been variable with both negative and positive results reported in the scientific literature
(Glaser at al., 2002). Several studies have reported that plant growth responses are largest when
charcoal and fertilizers are combined suggesting a synergistic relationship (Chan et al., 2007;
Lehmann et al., 2003; Steiner et al., 2007). Gundale and Deluca (2007) observed that laboratory
produced charcoal from ponderosa pine and Douglas-fir had a negative effect on plant growth
whereas the same charcoal created from wildfires showed a positive effect on plant growth. The
authors speculated that the low temperature charring method used to create the charcoal in the
laboratory either created toxic compounds that inhibited plant growth or acted as a source of
labile carbon (C) stimulating microbial growth and N immobilization. The objectives of the
present research were to determine the effects of charcoal volatile matter content on plant growth
and N transformations in a tropical acid soil. We hypothesized that biochar created at low
temperatures with high VM would increase microbial activity resulting in a decrease in plant
available N due to immobilization.

Western Nutrient Management Conference. 2009. Vol. 8. Salt Lake City, UT. Page 26
MATERIALS AND METHODS
Two greenhouse bioassays and two laboratory incubations were conducted to test the effects
of biochar VM content on plant growth and N transformations. The soil was an infertile, acid
Leilehua series (very-fine, ferruginous, isothermic, ustic kanhaplohumults) collected from the
30-80 cm depth at the Waiawa Correctional Facility, Mililani, Oahu Island (N21°26’53”, W157°
57’ 52”). The charcoal feedstock used in our experiments was macadamia nut shells. The
charcoal was made using a flash carbonization process developed at the Natural Energy Institute
at the University of Hawaii (Antal et al., 2003). Selected chemical properties of the soil and
biochars used in the different experiments are presented in Table 1. Total C and N content of the
biochars were determined by dry combustion on a LECO CN-2000. Biochar pH was measure in
1:1 slurry of charcoal to deionized water. Base cations were extracted with 1M ammonium
acetate at pH 7 and Al+++ was extracted with 1M KCl and measured in solution by inductive
coupled plasma spectrophotometer. The effective cation exchange capacity (ECEC) of the
biochars and soil was determined by summing the exchangeable cations.

Table 1. Selected chemical properties of the Leilehua soil, and the biochars used in the
greenhouse and laboratory experiments (LVM = low VM content and HVM = high VM
content).
VM Ash OC TN pH P K Ca Mg Na Al ECEC
% mg kg-1 cmolc kg -1

Soil
Leilehua 4.28 0.12 4.70 2.22 0.09 0.72 0.52 0.29 1.61 3.22
Charcoal
LVM
6.30 4.18 88.7 0.45 8.16 17.2 1.25 3.7 0.31 0.011 22.5
MacNut
HVM
22.5 3.33 85.2 0.45 5.72 18.5 0.74 0.7 0.15 0.032 20.2
MacNut

In the first greenhouse bioassay we imposed five treatments consisting of a control

(unamended soil), soil+lime, soil+biochar, soil+lime+NPK and soil+biochar+lime+NPK
arranged in randomized block design with four replications. The biochar contained 22.5 % VM
and was considered a high VM biochar. Biochar was applied to achieve 10% (w/w), lime to
achieve 2 T ha-1, N as NH4NO3 at a rate of 200 mg N kg-1, P as Ca(H2PO4)2 to achieve a rate of
750 mg P kg-1, K and Mg were added in solution at a rate equivalent to 200 and 100 kg ha-1
respectively, and the micronutrients Cu, Mn, and Zn were added in solution at a rate of 10 kg ha-
1
. We used corn (Zea mays, var super sweet #9) as the test crop. Eight corn seeds were planted
into each pot and thinned to four plants after emergence. The second greenhouse bioassay
consisted of five treatments (unamended soil, soil+lime+NPK, soil + high VM biochar, soil +
low VM biochar, soil + low VM biochar + NPK) installed in a complete randomized block
design with four replicates. Lime and fertilizers were applied at the same rates as in the first
experiment and corn was the test crop. At harvest time, above-ground biomass was cut at the soil
surface dried at 70°C for 72 hours, weighed and tissue analyzed for nutrient content according to
standard procedures (Hue et al., 2000).
We conducted two laboratory studies to evaluate the effect of biochar VM content on net N
mineralization rates and on CO2 respiration. Both experiments consisted of three treatments, a
control (untreated Leilehua soil) and the Leilehua soil amended with high and low VM
macadamia nut biochar applied at the same rate as in the greenhouse experiment. For the N

Western Nutrient Management Conference. 2009. Vol. 8. Salt Lake City, UT. Page 27
study, the biochar was mixed thoroughly with 50 g (oven dry equivalent) of soil followed by the
addition of the appropriate volume of deionized water required to bring the soil to 75% of water
holding capacity. The soils were placed in 100 mL beakers, weighed at the outset of the
incubation, covered with perforated parafilm, and incubated at constant temperature (28°C) and
moisture. Soils were sampled and analyzed for inorganic N, protease activity, and K2SO4
extractable organic C and TN after 2, 7, and 14 days. The soluble C fraction of the biochar was
determined by shaking 3 g of biochar in 30 mL deionized water for 1 hour and filtering through a
45 μm nylon membrane. For the CO2 respiration study, we used the alkali adsorption method
where 50 g of treated and untreated soils and 50 ml of 0.05 M NaOH were sealed in airtight 1 L
mason jars and incubated at 28°C for 14 days (Alef, 1995). The beaker containing the NaOH
solution was removed from the mason jar at 48 hour intervals and titrated with 0.05 M HCl
following the addition of 0.5 M BaCl2. Four mason jars with the 0.05 M NaOH solution, but
without soil were used as controls.

RESULTS AND DISCUSSION

The high VM biochar used in the first greenhouse bioassay had a significant negative effect
on corn growth compared to the control (Fig. 1). Amending the soil with conventional inorganic
fertilizers (lime+NPK) produced significant 5
increases in corn growth, but the beneficial
effects of the fertilizer was erased when a

4
combined with charcoal. Indeed, by
Shoot Biomass ( g pot-1)

combining charcoal with the fertilizer there

was an approximately 50% decline in corn 3

growth compared with the fertilized soil. b

Corn plants growing in the control soil 2

showed very low N, P and K concentrations c

in the tissue (data not shown). Tissue N and 1
c
d
K concentrations remained low after the
addition of charcoal, but P concentrations 0
S C L L L
S+ S+ F+ F+
increased significantly. Applying NPK S+
S+
C+

fertilizers significantly increased tissue N, P, Figure 1. Treatment effects on above ground corn dry
and K concentrations and the accompanying matter production in an infertile Leilehua soil amended
significant rise in dry matter production with high VM biochar and fertilizer (S = soil, S+C =
indicated that the Leilehua soil was severely soil + biochar, S+L = soil + lime, S+F+L = soil + NPK
deficient in N, P, and K. The biochar in + lime, S+C+F = soil + biochar + NPK).
combination with fertilizers, however,
significantly decreased tissue N, P, and K concentrations compared to the fertilizer control
treatment. Our observations were in disagreement with a recent greenhouse experiment reporting
that biochar significantly improved N fertilizer use efficiency by radish plants (Chan et al.,
2007). We speculated that the relatively high VM content of the biochar used in this experiment
may have played a role in inhibiting corn growth.

Western Nutrient Management Conference. 2009. Vol. 8. Salt Lake City, UT. Page 28
 The results of the second greenhouse experiment showed that biochar VM content had
significant effects on plant growth. High VM biochar significantly reduced shoot dry matter
compared with the control whereas low VM biochar had no significant effect on dry matter
production (Fig. 2). Corn growth was
significantly better in the low VM 0.4
b
charcoal treatment than in the high VM a

charcoal treatment. The low VM biochar

combined with fertilizer showed a 0.3

Shoot Biomass (g pot-1)

a
significant increase in dry matter
production compared with the fertilizer
alone treatment. The high VM biochar 0.2

reduced N uptake by 50% compared with

the control. On the other hand, the low 0.1
b
b
VM biochar did not reduce N uptake in
either biochar alone treatment or the c

biochar augmented with fertilizer 0.0

treatment. Although the low VM biochar S

S+H
VM
S+L
VM
S+N
PK+
L
S+L
VM
+NP
K

with fertilizer treatment did not show as

high an increase in plant growth nor a Figure 2. Treatment effects on above ground corn dry
significant increase in N uptake compared matter production in an infertile Leilehua soil amended with
high and low VM biochar and fertilizer (S = soil, HVM =
with the fertilizer treatment as in the high VM biochar, LVM = low VM biochar).
results reported by Chan and his group
(2007), our results provide evidence that the VM content of the biochar is an important factor
affecting its agronomic value as a soil amendment. We suspected that high VM charcoal is a
source of labile C for soil microorganisms, and the high C:N ratio of the C source stimulated
immobilization of the plant available N
in the soil causing N deficiency in the
growing plants. A recent experiment
50

reported similar results showing that

charcoal produced at low temperature 40

(350°C) had a negative effect on plant

NH4-N (mg kg )

growth (Gundale and DeLuca, 2007),

-1

30
and the researchers speculated that the
decline in plant growth was caused by 20
Soil
Soil+HVM
N immobilization due to high Soil+LVM

concentrations of soluble and total

phenols in the charcoal, which served 10

as a high C:N carbon source for soil

microorganisms. 0

Results from the two incubation 0 2 4 6 8 10 12 14 16

Days
experiments confirmed that biochar
VM exerts a strong influence on N +
mineralization and microbial Figure 3. Biochar effects on soil NH4 -N in a 14 - day
incubation.
respiration. The untreated soil showed
an initial drop in soil NH4+-N after two
days from 39.4 to 31.7 mg kg-1 followed by a slow increase to 45.3 and 43.4 mg kg-1 after seven

Western Nutrient Management Conference. 2009. Vol. 8. Salt Lake City, UT. Page 29
and fourteen days respectively (Fig. 3). The soil amended with high VM biochar, however,
showed a dramatic decline in soil NH4+-N that persisted throughout the fourteen day incubation.
The low VM biochar had a much smaller effect on soil NH4+-N decreasing it to around 30 mg
kg-1. In the CO2 respiration study, the high VM biochar amendment caused a steep increase in
respiration reaching a peak at four days followed by a gradual decline through the 12th day (Fig.
4). At day 2 and day 6 the high VM
biochar treatment showed a respiration 400

rate threefold higher than the control,

Soil
Soil+HVM
which remained at least twice as high as

Respiration (mg CO2 kg 48hr )

Soil+LVM

-1
300
the control throughout the remainder of

-1
the incubation period. The low VM
treatment showed an initial spike in 200
respiration at day 2 followed by a rapid
decline matching the control values by
the eighth day. The relatively high CO2 100

respiration rate combined with the

dramatic decline in soil NH4+-N
concentration observed in the high VM 0

biochar treatment is strong evidence that

0 2 4 6 8 10 12

Days
N immobilization by the microbial
biomass was an important factor Figure 4. Biochar effects on CO2 respiration in a 12-day
explaining the observed decline in plant incubation.
growth and N uptake in the high VM
biochar treatments. The high water extractable C content of the high VM biochar (265 mg C kg-
1
) compared with the low VM biochar (53 mg kg-1) provided a labile source of C fueling the
observed stimulation of microbial activity in the high VM treatment. With the high C:N ratio of
the biochar, the microbial biomass was forced to scavenge soil N inducing N deficiency in the
growing plants.

SUMMARY
This research shows that biochar VM content, or the degree of carbonization, can play a
critical role in determining its agronomic value as a soil amendment. Our results provide clear
evidence that biochars that are high in VM content (i.e., a typical barbecue charcoal) would not
be good soil amendments because they can stimulate microbial activity and immobilize plant
available N in the short-term. On the other hand, more fully carbonized biochars with lower VM
content containing a smaller labile C component have a smaller effect on soil microbial activity
and N immobilization. While our research provides one explanation for why some biochars have
a negative effect on plant growth, it still remains unclear why low VM biochars in combination
with fertilizer appear to have a beneficial effect on plant growth. Despite our findings elucidating
the role of VM content in inhibiting N mineralization, research at the field scale is required to
truly assess the agronomic value of biochars as soil amendments.

REFERENCES
Alef, K. 1995. Soil Respiration, p. 214-216, In K. Alef and P. Nannipieri, eds. Methods in
applied soil microbiology and biochemistry. Academic Press, London.

Western Nutrient Management Conference. 2009. Vol. 8. Salt Lake City, UT. Page 30
Antal, M.J., K. Mochidzuki, and L.S. Paredes. 2003. Flash carbonization of biomass. Industrial
& Engineering Chemistry Research 42:3690-3699.
Chan, K.Y., L. Van Zwieten, I. Meszaros, A. Downie, and S. Joseph. 2007. Agronomic values of
greenwaste biochar as a soil amendment. Australian Journal of Soil Research 45:629-634.
Glaser, B. 2007. Prehistorically modified soils of central Amazonia: a model for sustainable
agriculture in the twenty-first century. Philosophical Transactions of the Royal Society B-
Biological Sciences 362:187-196.
Glaser, B., E. Balashov, L. Haumaier, G. Guggenberger, and W. Zech. 2000. Black carbon in
density fractions of anthropogenic soils of the Brazilian Amazon region. Organic
Geochemistry 31:669-678.
Glaser, B., J. Lehmann, and W. Zech. 2002. Ameliorating physical and chemical properties of
highly weathered soils in the tropics with charcoal - a review. Biology and Fertility of Soils
35:219-230.
Gundale, M.J., and T.H. DeLuca. 2007. Charcoal effects on soil solution chemistry and growth
of Koeleria macrantha in the ponderosa pine/Douglas-fir ecosystem. Biology and Fertility of
Soils 43:303-311.
Hue, N.V., R. Uchida, and M.C. Ho. 2000. Sampling and analysis of soils and plant tissues. pp.
23-30, In J. A. S. a. R. S. Uchida, ed. Plant Nutrient Management in Hawaii Soils. College
of Tropical Agriculture and Human Resources, University of Hawaii, Honolulu.
Lehmann, J., J.P. da Silva, C. Steiner, T. Nehls, W. Zech, and B. Glaser. 2003. Nutrient
availability and leaching in an archaeological Anthrosol and a Ferralsol of the Central
Amazon basin: fertilizer, manure and charcoal amendments. Plant and Soil 249:343-357.
Sombroek, W.G., F.O. Nachtergaele, and A. Hebel. 1993. Amounts, dynamics and sequestering
of carbon in tropical and subtropical soils. Ambio 22:417-426.
Steiner, C., W. Teixeira, J. Lehmann, T. Nehls, J. de Macêdo, W. Blum, and W. Zech. 2007.
Long term effects of manure, charcoal and mineral fertilization on crop production and
fertility on a highly weathered Central Amazonian upland soil. Plant and Soil 291:275-290.

ACKNOWLEDGEMENTS
We thank Dr. Michael Antal for providing biochar samples along with proximate analysis
data and Yudai Tsumiyoshi and Jocelyn Liu for assistance with laboratory analysis. Funding for
this research came in part from USDA HATCH project 863H.

Western Nutrient Management Conference. 2009. Vol. 8. Salt Lake City, UT. Page 31
Copyeditor: Maria Timothy Santiago
TECHNICAL ARTICLE
Impact of Biochar Amendment on Fertility of a Southeastern
Coastal Plain Soil
Jeffrey M. Novak,1 Warren J. Busscher,1 David L. Laird,2 Mohamed Ahmedna,3
Don W. Watts,1 and Mohamed A. S. Niandou3
Abstract: Agricultural soils in the southeastern U.S. Coastal Plain
region have meager soil fertility characteristics because of their sandy
textures, acidic pH values, kaolinitic clays, low cation exchange capaci-
ties, and diminutive soil organic carbon contents. We hypothesized that
biochar additions will help ameliorate some of these fertility problems.
The study objectives were to determine the impact of pecan shellYbased
biochar additions on soil fertility characteristics and water leachate
chemistry for a Norfolk loamy sand (fine-loamy, kaolinitic, thermic typic
Kandiudults). Soil columns containing 0, 0.5, 1.0, and 2.0% (wt/wt)
biochar were incubated at 10% (wt/wt) moisture for 67 days. On days
25 and 67, the columns were leached with 1.2 to 1.4 pore volumes of
deionized H2O, and the leachate chemical composition determined. On
days 0 and 67, soil samples were collected and analyzed for fertility. The
biochar had a pH of 7.6, contained 834.2 and 3.41 g kgj1 of C and N,
respectively, and was dominated by aromatic C (58%). After 67 days and
two leaching events, biochar additions to the Norfolk soil increased soil
pH, soil organic carbon, Ca, K, Mn, and P and decreased exchangeable
acidity, S, and Zn. Biochar additions did not significantly increase soil
cation exchange capacity. Leachates contained increasing electrical con-
ductivity and K and Na concentrations, but decreasing levels of Ca,
P, Mn, and Zn. These effects reflect the addition of elements and the
higher sorption capacity of biochar for selective nutrients (especially Ca,
P, Zn, and Mn). Biochar additions to the Norfolk soil caused significant
fertility improvements.
Key words: Biochar, coastal plain soil, fertility, GRACEnet, leachate
(Soil Sci 2009;174: 00Y00)
F or more than 150 years, sandy soils of the southeastern U.S.
Coastal Plain region have been cultivated for row crops,
particularly corn and cotton (Gray, 1933; Trimble, 1974). Most
of these agricultural soils are Ultisols (Buol, 1973; Gardner,
1981) formed in fluvial and marine sediments (Daniels et al.,
1999) deposited 0.5 to 5 million years before present during
the Pliocene to early Pleistocene periods (Daniels et al., 1970;
1978). The warm, humid climate and long time for pedogenesis
have created distinct soil profile, mineralogical, and chemical
characteristics. Extensive clay eluviation has created shallow A
horizons (0Y0.20 m thick); well-developed E horizons (0.2Y1 m
1
USDA-ARSYCoastal Plain Soil, Water and Plant Research Center, Florence,
SC. Dr. Novak is corresponding author. E-mail: jeff.novak@ars.usda.gov
2
USDA-ARSYNational Soil Tilth Laboratory, Ames, IA.
3
Interdisciplinary Energy and Environment Program, North Carolina A&T
University, 171 Carver Hall, Greensboro, NC.
Received August 19, 2008, and in revised form November 30, 2008.
Accepted for publication December 2, 2008.
Manuscript accepted for publication to Soil Science on: September 24, 2008.
Mention of a specific product or vendor does not constitute a guarantee or
warranty of the product by the U.S. Department of Agriculture or imply
its approval to the exclusion of other products that may be suitable.
Copyright * 2009 by Lippincott Williams & Wilkins
ISSN: 0038-075X
DOI: 10.1097/SS.0b013e3181981d9a
Soil Science & Volume 174, Number 2, February 2009
Copyright @ 2009 by Lippincott Williams & Wilkins. Unauthorized reproduction of this article is prohibited.
thick) that have sand, loamy sand, or sandy loam textures; and
relatively thick Bt horizons (1.0Y3.0 m thick) with sandy clay to
clay textures (Daniels et al., 1970; Shaw et al., 2004). Intensive
leaching of bases has resulted in low soil pH values, extensive
clay mineral weathering, low cation exchange capacity (CEC)
values (2Y8 cmolc kgj1), and high levels of exchangeable Al
(Gamble and Daniels, 1974; Daniels et al., 1978). These soil
characteristics severely limit soil fertility and agricultural man-
agement options.
For instance, sandy soils exhibit high N leaching after fer-
tilizer or manure application (Trindale et al., 1997; Ritter et al.,
1998; Zotarelli et al., 2007). Moreover, Ultisols in the Coastal
Plain region have low soil organic carbon (SOC) contents in the
surface 0- to 15-cm depth (6.3Y9.2 g kgj1; Hunt et al., 1996;
Novak et al., 2007a) because of rapid residue oxidation, which is
further accelerated by inversion tillage for row crop production
(Bauer et al., 2006). The physical and chemical problems dis-
cussed above severely limit soil fertility and hence crop pro-
ductivity on the Ultisols of the southeastern U.S. Coastal Plain.
Fertility problems associated with southeastern Coastal
Plain Ultisols are similar to those of Oxisols in intertropical
regions (Eswaran and Tavernier, 1980), which also have low pH,
SOC, and CEC values (Tiessen et al., 1994; Lehmann et al.,
2003). Poor soil fertility raises concerns about the sustainability
of agriculture in regions dominated by Oxisols and has spurred
the development of management practices to restore or im-
prove their fertility status (Glaser et al., 2002). Applications of
mulches, composts, and manures increase soil fertility; however,
under tropical conditions, the increase is short term because the
added organic matter is quickly oxidized and added bases are
rapidly leached (Tiessen et al., 1994).
On the other hand, application of biochar (charcoal pro-
duced by pyrolysis of biomass feedstock) to infertile Oxisols has
been shown to provide longer-lasting improvements in soil fer-
tility (Glaser et al., 2002; Lehmann et al., 2003; Steiner et al.,
2007). Biochar composed primarily of single and condensed
ring aromatic C (Lehmann, 2007) has both a high surface area
per unit mass and a high charge density. Because of these
properties, biochar is both more recalcitrant in tropical soils
and contributes a higher capacity to sorb cations per unit mass
than does biogenic soil organic matter (Sombroek et al., 2003;
Liang et al., 2006).
Biochar application to soils is not a new concept (Mann,
2005). For example, in the Amazon basin, anthropogenic dark
earth soils (referred to as Terra Preta) contain large amounts of
charred materials most likely added by pre-Columbian farmers
who practiced a form of slash and char agriculture (Sombroek
et al., 2003) along with disposal of charcoal remains from hearths
(Glaser et al., 2002). In these soils, the biochar acts as a soil con-
ditioner, improving soil physical properties and nutrient use effi-
ciency, thereby increasing plant growth. Today, 500 years after
cessation of the practices that created these soils, the Terra Preta
soils are highly valued for agricultural and horticultural use in the
Amazon basin (Glaser et al., 2002; Lehmann and Rondon, 2006).
1
 Novak et al
To predict the reactivity as well as stability of biochar when
used as a soil amendment, it is important to know the biochars
organic structural composition. Biochar from Terra Preta soils
analyzed using solid-state nuclear magnetic resonance (NMR)
analysis was shown to be composed of a highly heterogeneous
mixture of organic structures (Novotny et al., 2007). The struc-
tural form of C in biochar depends on the biogeochemistry of the
biomass feedstock and the conditions under which it was py-
rolyzed (Kramer et al., 2004; Lehmann, 2007). Biochars com-
posed primarily of condensed aromatic C are known to persist in
soil environments for millennia, whereas biochars with higher
levels of single-ring aromatic and aliphatic C will mineralize
more rapidly (Lehmann, 2007; Novotny et al., 2007). Surface
area and surface charge density of biochar will have a large
influence on soil CEC and the ability of biochar additions to
ameliorate soil fertility problems.
We hypothesize that biochar additions to the sandy, Coastal
Plain soils of the southeastern United States would increase
the SOC content and CEC and improve the fertility status. The
specific objectives of this investigation were to determine the 1)
chemistry of biochar derived from pyrolysis of pecan shells, a
locally abundant source of feedstock; 2) effects of biochar addi-
tions on fertility characteristics of a Norfolk soil; and 3) effects
of biochar additions on the chemical composition of leachate
collected from a Norfolk soil.
MATERIALS AND METHODS
Production and Characterization of Pecan
ShellYBased Biochar
Pecan shells were obtained from a supplier in Lumberton,
NC. They were ground using a Retsch Mixer Mill (SR-2000;
Cole-Palmer, Vernon Hills, IL) to pass through a 2-mm sieve.
Per each pyrolysis batch, approximately 1000 to 2000 g of shells
were placed into a crucible (25 cm wide ! 10 cm deep) and were
inserted into a Lindberg box programmable furnace equipped
with an airtight retort (model 5116HR; Lindberg, Watertown,
WI). The furnace retort atmosphere was purged with N2 using
a flow rate of 0.1 m3 hj1. The furnace was controlled with a
multiple-step pyrolysis temperature program. The furnace was
initially heated to 40 -C; temperature was ramped to 170 -C at
5 -C minj1 and was maintained at this temperature for 30 min.
The temperature was then ramped to 700- C at 5 -C minj1,
and the pecan shells were subjected to pyrolysis for 1 h. The
biochar was cooled in the oven under the N2 atmosphere over-
night. After cooling, the biochar was ground to pass through a
0.25-mm sieve.
The sieved biochar moisture percent (wt/wt) content was
measured by oven drying a 2-g portion overnight at 80 -C.
Biochar pH was measured according to Ahmedna et al. (1998).
The method consisted of preparing a 1% (wt/wt) suspension of
biochar in deionized water. The suspension was heated to about
90 -C and stirred for 20 min to allow dissolution of the soluble
biochar components. After cooling to room temperature, the pH
of the biochar suspension was measured using a Corning pH
meter (Acton, MA). The biochar percent ash content (wt/wt) was
determined by dry combustion at 760 -C in air for 6 h using an
AQ1 Isotemp laboratory muffle furnace (Fisher Scientific). Biochar
percent moisture was 1.4%, pH was 7.5, and ash content was
3.8% (wt/wt).
A sample of the biochar was analyzed for total C, H, N, S,
and O (by difference) determination using ASTM D 3176
method (ASTM, 2006). For total elemental analyses, the biochar
was redried and then digested using the EPA method 3052
microwave-assisted acid digestion method (USEPA, 2008). The
2 * 2009 Lippincott Williams & Wilkins
Copyright @ 2009 by Lippincott Williams & Wilkins. Unauthorized reproduction of this article is prohibited.
Soil Science & Volume 174, Number 2, February 2009
elemental concentrations (Ca, Cr, Cu, Fe, K, Mg, Mn, Na, Ni, P,
Si, and Zn expressed on a dry, wt/wt, ash-free basis) in the
biochar digests were measured using an Elan DRC-II (Perkin-
Elmer, Shelton, CT) inductively coupled plasma mass spec-
trometer. Analytical operating conditions and element detection
limits are available at EBG-SWES-UA (2008).
Solid-state cross-polarization magic angle-spinning total-
sideband suppression 13C NMR spectral pattern of the biochar
was obtained using a Bruker DSX-300 spectrometer (Karlsruhe,
Germany) operated at a 13C frequency of 75.5 MHz. Additional
technical parameters to acquire the spectra have been described
by Wang et al. (2007). The chemical shift region assignments
were as follows: 0 to 50 ppm, aliphatic C; 50 to 109 ppm,
O-alkyl C; 109 to 163 ppm, aromatic C; and 163 to 190 ppm,
carboxylic C. The percent C distribution was determined by
estimating the area in these chemical shift regions as a per-
centage of the total area under the spectral curve.
Norfolk Soil Collection and Analyses
A bulk sample of a Norfolk soil from the Ap horizon
(0Y15 cm deep) was collected from a field contiguous to the
USDA-ARSYCoastal Plain Research Center, Florence, SC. The
field is nearly level (1Y2% slopes) and has a 30-year history of
row crop production (Sojka et al., 1984). The soil was collected
using a shovel in mid-April, approximately 1 week after fertili-
zation with 49 kg N haj1 of 28-0-0 UAN (urea + NH4NO3) for
an upcoming corn crop. The soil was air dried and 2-mm sieved.
The Ap horizon is a loamy sand with a particle size distribution
of 730, 250, and 20 g kgj1, respectively, of sand, silt, and clay
(sedimentation method; Soil Characterization Lab, Ohio State
University, Columbus, OH). X-ray diffraction analysis of the
clay fraction revealed a preponderance of kaolinite, with minor
amounts of hydroxy (Fe and Al) interlayered chlorite (X-ray
diffraction method; Soil Characterization Lab, Ohio State Uni-
versity). The pH of the untreated Norfolk Ap soil was 4.8, as
measured in a 1:1 soil-to-deionized water mixture (Novak et al.,
2007a). The total C and total combustible nitrogen (TCN)
contents were determined using a LECO TruSpec CN analyzer
(LECO Corp., St Joseph, MI). Soil C was assumed to be organic
in nature because the low soil pH precluded carbonates.
Biochar Incubation in Norfolk Ap Soil
The biochar incubation experiment was conducted in open-
top, 10-cm-diameter, 17-cm-tall schedule-40 PVC columns.
Column bottoms were sealed using a nylon mesh fabric to
support the soil bed and minimize soil loss. Sufficient amounts
of 0.25-mm sieved biochar was mixed into 750-g of air-dried,
2-mm-sieved Ap horizon soil to create 0, 0.5, 1.0, and 2.0%
(wt/wt) biochar treatments. These biochar rates equated to field
applications of approximately 0, 10, 20, and 40 metric tons haj1.
Each treatment was set up in triplicate. Deionized H2O was
then mixed into each treatment to obtain a soil-moisture content
of 10% (wt/wt), representing the upper range (between 5 and
10%) of field capacity for a typical Norfolk Ap soil horizon. The
moist soil treatments were then placed into columns, and they
were tamped down by hand to obtain a bulk density of 1.2 g cm3.
This created a headspace above the soil of 8 to 10 cm for adding
water. The columns were laboratory incubated for 67 days at 10%
soil moisture. The laboratory room temperature and percent
relative humidity, respectively, throughout the incubation ranged
between 17 and 27 -C and 23 and 61%.
On days 25 and 67, each column was leached with 1.2 to 1.4
pore volumes of deionized water; the leachate was collected and
weighed. The leachates were analyzed for total organic carbon
(TOC) concentrations using a Shimadzu TOC-Vcs (Shimadzu
 Soil Science & Volume 174, Number 2, February 2009 Biochar in Coastal Plain Soil

FIGURE 1. 13C NMR spectral pattern for pecan shellYbased biochar (%C distribution for structural groups determined as described by
Wang et al. (2007).

Corp., Columbia, MD), and for Ca, Cu, Fe, K, Mg, Mn, Na, P, S, structures of the biochar. This speculation has merit because
and Zn concentrations with a Varian ICP-OES (Varian Inc., Palo the high pyrolysis temperature explains the lack of alkyl C
Alto, CA). The ICP detection limit for this suite of 11 elements (0Y50 ppm) as volatile material such as oils, fatty acids, and
was a conservative 0.05 mg Lj1. The leachate pH and electrical alkyl alcohols would be lost (Antal and GrLnli, 2003). Carboxyl-
conductivity (EC) were measured using a standard pH and a containing structures were present in the NMR spectra possibly
conductivity meter. because of their structural decomposition resistance during
Samples of the biochar-treated Norfolk Ap soil were col- pyrolysis. Alternatively, their presence could be due to water
lected on incubation days 0 and 67 for analysis of plant available sorption during handling and grinding. The biochar percent
nutrients using Mehlich 1 (diluted HCl and H2SO4) extractant. moisture after grinding and before total elemental analyses was
Measurements of Mehlich 1Yextractable Ca, Cu, Fe, K, Mg, less than 2% and 4%, respectively. Consequently, single-ring
Mn, Na, P, S, and Zn and exchangeable acidity values were aromatic and some heterocyclic compounds could have been
conducted by the Clemson University Soil Testing Laboratory reoxidized, forming carboxylic and acetyl OH groups as evident
using ICP. The pH and EC of the biochar-treated Norfolk Ap
soil were measured using the methods of Novak et al. (2007a).
Statistics TABLE 1. Total elemental analyses of pecan shellYbased
The mean values of SOC and TCN contents between the biochar and Norfolk Ap soil† (data sorted into macroelement,
treatments, sorted by incubation day, were tested using a one- microelement pools)
way analysis of variance (ANOVA) with a P G 0.05 level of
Biochar Norfolk Ap soil
significance. The soil fertility characteristics and chemical com-
position of the deionized water leachates were sorted and tested Macroelement (g kgj1)
in a similar manner. All statistical tests were performed using Al 0.22 5.1
SigmaStat v. 3.5 software (SSPS Corp., Chicago, IL). C 834.2 16.8
Ca 3.64 0.49
RESULTS AND DISCUSSION Fe 0.07 2.91
Biochar and Soil Compositional Analyses H 10.3 V
F1 The 13C NMR spectral pattern of the pecan biochar (Fig. 1) K 4.15 0.38
revealed two prominent peaks at 120 and 187 ppm and a N 3.41 1.26
shoulder near 90 ppm. These peaks indicate that most of this O 19.8 V
biochar is distributed in aromatic structures (58%), with less Si 104.9 424.8
amounts of C having single bonds to O (29%) and in carboxyl Microelements (mg kgj1)
(13%) groups. The lack of a distinct peak near 75 ppm suggests Cu 14 4.6
little carbohydrate C; rather, the shoulder near 90 ppm is more Cr 0.31 23
characteristic of acetal C (Wershaw, 1985). The pecan shells Mg 698 445
used as feedstock for preparation of the biochar are composed
Mn 78 55
primarily of lignin and cellulose (47%, acid-detergent lignin;
Na 218 951
Ramirez et al., 1986). Charring of lignin and cellulose at tem-
peratures of 500 -C was reported to cause loss of their aliphatic Ni 0.5 2
components along with a conversion of ring structures into P 263 185
aromatic compounds (Rutherford et al., 2004). During pyrolysis S 95 2
at 700 -C in this study, it was suspected that a similar decom- Zn 7 43
position of cellulose and lignin and structural rearrangement of †
Determined on biochar and soil using EPA 3052 method (HNO3 + HF).
ring compounds to form condensed and single-ring aromatic

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 Novak et al Soil Science & Volume 174, Number 2, February 2009

sand-size fraction (Smith et al., 1976), and Fe-oxides and hy-

TABLE 2. SOC and TCN contents of Norfolk Ap soil and
soil + percent biochar mixtures on 0- and 67-day incubations
SOC (g kgj1)† TCN (g kgj1)
Norfolk Ap
soil + % biochar 0d 67 d 0d
a a a a
0 17.0 17.4 1.26
0.5 18.1a 18.3a 1.14a
1.0 22.2b 21.9b 1.25a
2.0 31.2c 29.2c 1.49b
†
Means compared within a column followed by a different letter are
significantly different at P G 0.05 using a one-way ANOVA (multiple
comparisons vs. Norfolk Ap soil + 0% biochar as a control).
in the 13C NMR spectra (Fig. 1). The NMR spectra indicate that
this pecan shellYbased biochar is composed of a mixture of
organic structural groups reflecting the chemistry of the feed-
stock and reactions occurring during both pyrolysis and after
pyrolysis on exposure of the biochar to oxygen and water. These
findings are similar to reports for black C isolated from dark
earth soils (Schmidt and Noack, 2000; Novotny et al., 2007).
The total elemental analyses of the pecan shell biochar and
T1 the Norfolk Ap soil were quite dissimilar (Table 1). The biochar
was enriched in C, Ca, K, Mg, N, and Si compared with soil,
whereas the Norfolk Ap soil was enriched in Al, Fe, Na, and Si.
Pyrolysis of organic feedstock’s from 400 to 700 -C results in a
concentration of C but a reduction of O and H due to evaporation
of sorbed H2O and driving off of jOH functional groups (Antal
and GrLnli, 2003). The N-containing structures in the biochar,
such as amino acids, amines, and amino sugars during the high-
temperature pyrolysis (700 -C) process were probably condensed
to form N-heterocyclic aromatic structures (Koutcheiko et al.,
2007). Thus, much of the residual N in the biochar (Table 1) was
likely present as recalcitrant heterocyclic N rather than the more
bioavailable amine N.
The elemental composition (C, H, O, N, and S) of biochar
was similar to values reported for other carbonized charcoals
(Antal and GrLnli, 2003). The Norfolk Ap soil elemental com-
position was Al-, Fe-, and Si-enriched; these elements are pre-
dominant in the chemical structure of aluminosilicates in the
droxides in the clay-size fraction (Shaw et al., 2004).
Changes in Soil Organic Carbon Content and
Fertility Characteristics
67 d The Norfolk Ap soil treatments with 1 and 2% biochar
addition on day 0 had significantly greater mean SOC contents
1.24
than the control (Table 2); mean SOC content was increased T2
1.28a
between 5.1 and 14.2 g kgj1. Similar SOC concentrations were
1.09b present in the soil after 67 days of incubation, indicating no
1.21a significant loss of biochar C during the incubation. The biochar
contained some N (3.41 mg kgj1; Table 1); however, mixing
0.5 to 1.0% biochar had no detectable effect on TCN of the
Norfolk Ap horizon. Adding 2% biochar significantly increased
the soil mean TCN content, but only by 0.23 g kgj1. This trend
was not observed after 67 days of incubation. Mean TCN
contents in treatments after 67 days of incubation (except 1.0%
biochar) were similar to the control (0% biochar).
The C:N ratio of the pecan biochar is 244:1. Nitrogen im-
mobilization typically occurs when organic residues possessing
a C:N ratio of greater than 32:1 are added to soils (Alexander,
1977; Thompson and Troeh, 1978). The wide C:N ratio, in
association with its aromaticity, will cause slow biochar de-
composition (Lehmann, 2007). Although biochars/soil black
carbon will undergo slow chemical and microbial decomposi-
tion (Schmidt and Noack, 2000), the rate of decomposition is so
slow that even large additions of biochar to soil will probably not
significantly immobilize N.
The high stability of biochar in soil environments is bene-
ficial with respect to C sequestration because C added to the soil
as biochar will be removed from the atmosphere for 1000 years
or more. German (2003) reported that biochar in soils is stable
and resistant to microbial attack; one site in the Amazonian
Black Earth region had biochar dated to 6850 years old. In com-
parison, the mean residence time of soil organic matter has been
estimated as between 250 and 3280 years (Stevenson, 1994). In
hind sight, we suggest that the SOC dated in those presented in
Stevenson (1994) likely contained significant amounts of black
C, which would make the average age of the total SOC pool much
older than the age of the biogenic SOC fraction. Laird et al.
(2008) physically separated biogenic humic material from black
C from an Iowa Mollisol and reported modern radio C dates for

TABLE 3. Fertility characteristics of Norfolk Ap soil + percent biochar mixtures on 0- and 67-day incubations
(Mehlich 1 extractant)†

Fertility characteristics‡
§
Incubation pH CEC Exch. acid Ca Cu K Mg Mn Na P S Zn
Norfolk Ap
j1 j1
soil + % biochar day - - - - - -cmolc kg ------- - - - - - - - - - - - - - - - - - - - - - - mg kg ----------------------
a a a a a a a a
0 0 4.8 5.7 2.4 437 0.6 35 117 12 5a 30.5a 8.67a 13a
0.5 0 5.1b 5.3a 2.0a 470b 0.7a 49b 98b 7b 5a 30.8a 7.67a 12a
1.0 0 5.5c 5.4a 1.9a 516c 0.7a 66c 90c 15c 6a 31.2a 7.83a 11a
2.0 0 6.3d 5.9a 1.2b 720d 0.8a 111d 91d 10d 7b 35.2b 8.50a 11a
0 67 5.2a 5.2a 2.4a 392a 0.6a 26a 93a 7a 3a 28.7a 6.33a 12a
0.5 67 5.6b 5.4a 2.1a 462b 0.7a 47b 91a 6a 5b 31.7b 5.16a 11b
1.0 67 5.9c 5.6a 2.0a 537c 0.7a 49c 92a 16b 4a 31.7c 4.00b 11b
2.0 67 6.4d 5.9a 1.5b 692d 0.8a 69d 89a 10c 4a 33.3d 3.17c 10c
†
Extracted with H2SO4 + HCl.
‡
Mean values sorted by incubation day were compared using a one-way ANOVA for multiple-comparisons tests vs. a control (Norfolk Ap soil +
0% biochar).
§
Means followed by a different letter are significantly different at P G 0.05.

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 Soil Science & Volume 174, Number 2, February 2009
the biogenic humic material and much older dates for the black
C fraction. Therefore, soil applied biochar has the potential to
increase the recalcitrant pool of soil C and will persist in soil
environments much longer than C added in the form of residues
or biogenic soil organic matter.
Biochar additions to soils are reported to improve soil fer-
tility by raising soil CEC (Liang et al., 2006). Soil CEC increases
are due to carboxylate groups on the surfaces of the biochar itself
and to exposed carboxylate groups of organic acids sorbed by the
biochar, both of which contribute negative surface charge to
biochar particles (Liang et al., 2006). Biochar in this experiment
contained some carboxyl characteristics (13%; Fig. 1); yet, the
ability of biochar additions to increase the soil CEC even at 2%
T3 biochar addition was negligible (Table 3). One might expect after
a 67-day incubation that additional carboxylate groups would
form because of oxidation of the biochar surfaces (Schmidt and
Noack, 2000). However, differences in Norfolk Ap CEC values
for the day-0 and day-67 samples were negligible (Table 3).
The high pyrolysis temperature (700 -C) may have con-
tributed to the relatively low level of surface oxidation of the
pecan shell biochar and hence the lack of a significant impact of
biochar additions on the CEC of the Norfolk Ap horizon soil.
Higher pyrolysis temperatures generally cause greater conden-
sation of aromatic structures and even the formation of graphitic
cores (Antal and GrLnli, 2003). Such highly condensed aromatic
C has less surface area and fewer oxidizable surface functional
groups than more open (less condensed) aromatic C structures.
High-temperature biochars are also more resistant to chemical
oxidation and microbial degradation and hence have a longer
half-life in soil environments than soil organic matter. The re-
calcitrant characteristics of high-temperature biochar, however,
would be a desirable property if the primary goal was to remove
atmospheric CO2 and sequester C in soil for millennia (Laird,
2008). On the other hand, if the primary goal was to increase soil
CEC values, then the addition of biochar prepared by pyrolysis
of feedstocks at lower temperatures (400Y500 -C) or under
different moisture and pressures conditions (Antal and GrLnli,
2003) may be more desirable. Low-temperature biochars will
most likely also increase soil C sequestration, but they will prob-
ably more rapidly change soil fertility characteristics when com-
pared with using high-temperature biochars.
Some microbial oxidizable compounds such as anhydrocel-
lulose (dehydrated forms of cellulose), polysaccharides, alcohols,
and so on, should exist in biochars prepared by pyrolysis of
feedstocks at lower temperatures (Antal and GrLnli, 2003).
Baldock and Smernik (2002) and Hamer et al. (2004) both
reported a relationship between biochar pyrolysis temperature
and resistance to soil microbial decomposition. Hamer et al.
(2004) found that biochars produced from maize and rye at
350 -C were more prone to soil microbial degradation than
biochar made from oak wood pyrolyzed at 800 -C. The authors
attributed differences in biochar decomposition because of their
C:N ratios; higher pyrolysis temperatures caused wider C:N
ratios in the oak wood biochar because of loss of N and con-
centration of C (Hamer et al., 2004).
Upon decomposition and oxidation by soil microbial com-
munities, these organic carbon structures should produce by-
products containing a higher density of carboxylate and other
O-containing functional groups (i.e., jOH, jOR, etc.) capable of
serving as sites for cation exchange (Stevenson, 1994). Therefore,
when creating biochars for use as a soil fertility amendment, the
biomass pyrolysis conditions could be designed to carbonize the
material under moist conditions and at lower temperatures.
Research has shown that soil pH is more influenced by
monomeric Al species on exchange sites than by H+1 (Sparks,
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Biochar in Coastal Plain Soil
1995). Aqueous monomeric hexahydronium [Al(H2O)6]+3 spe-
cies act as pH buffers because they can undergo rapid and
reversible hydrolysis reactions influencing solution pH values
by liberating or accepting H+1 (Sparks, 1995). For soil pH to
change, the biochar itself or a cation in the biochar must react
with the soluble monomeric Al species or displace it from ex-
change surfaces on clays or soil organic matter. At pH 4.8, the
Norfolk soil with no biochar had 42% (2.4 cmolc exchangeable
acidity kgj1 " 5.7 cmolc CEC kgj1 ! 100; Table 3) of the total
soil CEC sites occupied by [Al(H2O)5]+2. This would be the
dominant monomeric Al species in the Norfolk Ap soil + 0%
biochar at pH 4.8 (Sparks, 1995). Additions of 0.5 and 1%
biochar to the Norfolk soil did not significantly modify the ex-
changeable acidity values, although soil pH values significantly
increased by 0.7 U (Table 3). With the addition of 2% biochar,
the pH increased from 4.8 to 6.3, and exchangeable acidity was
reduced by 50% ($1.2 cmolc kgj1; Table 3). Thus, biochar was
an effective liming agent, neutralizing solution pH and reducing
exchangeable acidity values. However, substantial additions of
biochar (2% or 40 metric tons haj1) were required to obtain in-
creases in pH and reductions in exchangeable acidity.
During pyrolysis, cations (primarily K, Ca, Si, and Mg) pre-
sent in the pecan shells formed metal oxides (e.g., ash) that were
admixed with the biochar. Once in the soil environment, these
oxides can react with H+1 and monomeric Al species, modifying
soil pH and exchangeable acidity values. Because the biochar
contained a high Ca concentration (3.64 mg kgj1; Table 1),
Reaction (1) involving CaO exemplifies the liming ability of the
ash associated with the biochar:
2Al # soil þ 3CaO þ 3H2O Y 3Ca # soil þ 2A1ðOHÞ3 ð1Þ
During this reaction, Ca replaces the monomeric Al species
on soil mineral or soil organic matter CEC sites. Accompanying
this reaction is an increase in soil solution pH caused by the
depletion of the readily hydrolysable monomeric Al and the for-
mation of the more neutral [Al(OH)3]0 species (Sparks, 1995).
This general reaction explains the decline in exchangeable
acidity for the Norfolk Ap soil and the increase in solution pH
and Ca on CEC sites (Table 3). The pH increase and exchange-
able acidity decrease were similar for the day-0 and day-67
samples. This suggests that the liming effect of biochar occurred
rapidly and was sustainable on equilibration.
The biochar significantly increased some important plant
macronutrients. Mehlich 1Yextractable Ca, K, and P all increased
with the level of biochar additions (Table 3). However, the P
increase was significant only at the highest level of biochar ad-
dition. By contrast, Mehlich 1Yextractable Mg decreased with
increasing biochar addition at day 0, suggesting that the Mg was
strongly retained by the biochar. Extractable S and Zn concen-
trations also decreased slightly with increasing biochar additions,
but the trend was significant only for the day-67 samples. Copper
concentrations were not significantly affected by biochar addi-
tions, and Mn concentrations were variable. The observed varia-
tions in Mehlich 1Yextractable plant nutrient concentrations, at
days 0 and 67, reflected the combined effects of fertilization
(nutrients added with the biochar), leaching of nutrients, and
nutrient adsorption by the soil and added biochar. No plants were
grown in these soils, and hence, plant uptake was not a variable in
this study. In general, biochar additions increased the levels of
plant macronutrients and had little effect on micronutrients.
Chemical Composition of the Water Leachates
It is important to examine the chemical composition of a
deionized water extract of an amended soil; in some cases, the
amendment can release elements that may cause plant growth
5
 Novak et al Soil Science & Volume 174, Number 2, February 2009

TABLE 4. Chemical composition of deionized water leachates from Norfolk Ap soil + percent biochar mixtures after 25- and
67-day incubation†

Leachate chemical composition‡

§
Incubation pH EC TOC Ca Cu K Mg Mn Na P S Zn
Norfolk Ap
j1 j1
soil + % biochar day KS cm - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - mg L -------------------------
0 25 5.7a 364a 76a 189a V 32a 113a 5.4a 8.2a 2.0a 25a 1.3a
0.5 25 6.0a 382a 65b 175a V 58b 82b 1.1b 11.0b 1.4b 22a 0.7a
1.0 25 6.0a 439a 70a 136b V 72c 63c 0.3c 11.9c 1.5c 23a 0.4a
2.0 25 6.0a 559a 82a 99c V 99d 34d 0.2d 14.0d 1.2d 26a 0.1b
0 67 5.8a 364a 45a 31a V 9a 18.6a 0.1 2.80a 2.20a 9.5a 0.27a
0.5 67 5.9a 387a 38a 32a V 17a 17.1a V 3.02a 1.26b 8.9a 0.12b
1.0 67 5.7a 502a 48a 37a V 25b 17.1a 0.1 4.02a 1.40c 10a 0.13c
2.0 67 6.0a 571a 68a 46a V 52c 15.9a V 5.90b 1.42d 11a 0.06d
A dash line indicates mean value was below detection limit (0.05 mg Lj1).
†

‡
Mean values sorted by incubation day were compared using a one-way ANOVA for multiple comparisons vs. a control (Norfolk Ap soil +
0% biochar, significant digits varied because of low mean element concentrations on day 67).
§
Means followed by a different letter are significantly different at P G 0.05.

issues (Novak et al., 2007b). For the 25- and 67-day leachates, pH, combination of reactions such as retention of o-PO4j3 through
EC values, and TOC concentrations (except 0.5% on day 25) were ligand exchange reactions involving O-containing functional
T4 similar for the biochar-treated soils and control soils (Table 4). groups on the biochar surface, adsorption of o-PO4j3 by Fe and
The cation concentrations in the 25- and 67-day leachates Al oxides and hydroxides, and by adsorption and precipitation by
seem to be influenced by cation valency. For instance, the Ca, Mg-phosphates (Bohn et al., 1979). Regardless of sorption
monovalent cation (K and Na) concentrations in both the 25- and mechanisms, these results suggest that biochar has the potential
67-day leachates increase with the level of biochar addition to to ameliorate P leaching in soils with sandy textures, a common
the Norfolk Ap soil, whereas concentrations of multivalent problem in fields containing excess soil P concentrations as a
cations (Ca, Mg, Mn, and Zn) all decreased for the day-27 result of overapplication of swine manure (Novak et al., 2000).
leachates and either decreased or were not significantly different
for the day-67 leachates with increasing levels of biochar ad- Net Norfolk Ap Soil Fertility Changes
dition. The strength of cation retention or repulsion from nega- The net soil fertility change to the biochar-treated Norfolk
tively charged surfaces increases with increasing ion charge and Ap soil is presented in Table 5. The results after adding 2% T5
with distance between the charged surface and either the source biochar were presented because this treatment caused the most
of charge or the soluble ion (basis of diffuse double layer theory; significant soil fertility variations compared with the untreated
Bohn et al., 1979). Consequently, multivalent cations were pref- soil. The treated Norfolk Ap soil pH was more basic after
erentially adsorbed over monovalent cations on exchange sites,
and hence, the monovalent K and Na cations would be more
available for movement with the leachate. TABLE 5. Changes in Norfolk Ap soil fertility characteristics
The decrease in concentration of multivalent cations in the with and without 2% biochar (after 67-day incubation and
two deionized water leaching events)
leachates with increasing levels of biochar addition (Table 4)
is particularity interesting in light of the fact that substantial 0% 2% Net Relative
amounts of Ca and Mg and less amounts of Mn and Zn were Characteristic Unit Biochar Biochar change %†
present in the biochar (Table 1). Much of the Ca added with
the biochar probably replaced monomeric Al species on clays pH V 4.8 6.4 +1.6 +33
and/or soil organic matter exchange sites [see Reaction (1)]. CEC cmolc kgj1 5.7 5.9 +0.2 +3.5
This hypothesis is supported by increasing levels of Mehlich Exchange cmolc kgj1 2.4 1.5 j0.9 +38
1Yextractable Ca with higher levels of biochar addition (Table 3). acidity
An explanation for the decrease in Mg, Mn, and Zn concentra- C g kgj1 17.4 29.2 +11.8 +68
tions in day-27 leachates is a bit more complex, because their Ca mg kgj1 437 692 +255 +58
Mehlich 1Yextractable concentrations of these elements either Cu mg kgj1 0.6 0.8 +0.2 +33
decreased or did not show substantial changes with increasing K mg kgj1 35 69 +37 +106
biochar additions. These observations suggest that the Mg, Mn, Mg mg kgj1 117 89 j28 j24
and Zn were either specifically adsorbed or very highly selec-
Mn mg kgj1 12 10 j2 j17
tively adsorbed by exchange sites associated with the biochar.
N g kgj1 1.24 1.21 j0.03 j2.4
The P concentrations generally decreased with increasing
biochar application in both 25- and 67-day leachates by approxi- Na mg kgj1 5 4 j1 j20
mately 40% (0 vs. 2% biochar addition; Table 4). The field from P mg kgj1 31 34 +3 +10
which the Norfolk soil was collected has a long history of row S mg kgj1 9 4 j5 j56
crop production; past P fertilizer applications were the likely Zn mg kgj1 13 10 j3 j23
source of much P in the soil. The declines in leachate P concen- †
Percent change based on values in Norfolk Ap soil + 0% biochar.
trations with increasing biochar additions are probably due to a

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Soil Science & Volume 174, Number 2, February 2009
biochar addition through a 38% reduction in monomeric Al
species on exchange sites (lower exchangeable acidity values).
Soil Ca and K concentrations had marked increases (+58 and +
106%, respectively) implying that biochar additions can increase
these important plant macronutrients.
The Norfolk Ap horizon had an 11.8 g C kgj1 increase
after mixing in 2% biochar, and there was no detectable loss of
SOC during the 67-day incubation, suggesting that biochar ad-
ditions may be an effective means of sequestering C; longer-term
studies need to be quantified in terms of C sequestration. Two
percent biochar application to field soil is approximately 40 tons
biochar haj1. This is a large amount of biochar to apply to soil,
but the benefits of long-term C sequestration coupled with ad-
ditions of several plant nutrients, neutralizing soil acidity, and
reduced nutrient leaching should not be overlooked. Although
we observed decreased leaching of several plant nutrients with
increasing biochar additions, we did not observe significant
increases in CEC or EC for the Norfolk Ap soil. The influence of
biochar additions on Mehlich 1Yextractable micronutrient con-
centrations was minimal.
CONCLUSIONS
Biochar has been used as a fertility amendment in soils of
tropical regions for thousands of years, although scientific in-
vestigations of the effects on soil fertility are few. This same
technology may improve fertility-poor soils in the southeastern
U.S. Coastal Plain region. A laboratory study was conducted
where a pecan shellYbased biochar was incubated in a sandy,
acidic Norfolk Ap soil. Biochar additions increased the SOC
content but did not significantly improve the soil N status.
Biochar additions seem to potentially be an effective means of
sequestering C in soils as no detectable loss of SOC occurred
during the 67-day incubation. A goal of this study was to in-
crease the Norfolk Ap soil CEC by adding biochar. For soil
CEC to increase, surfaces of the biochar must be oxidized to
produce negatively charged carboxylate groups. No increase in
soil CEC after 67 days was obtained, implying that this high-
temperature biochar was not suited for that purpose within the
time frame of this study. Soil pH and three major plant nutrients
(Ca, K, and P) concentrations, however, increased after applying
this biochar. Most soil micronutrient concentrations were not
influenced by the biochar additions. Water leaching of biochar-
treated Norfolk Ap soil showed K enrichment but net sorption of
P and most multivalent cations.
This biochar was highly recalcitrant because the pecan
shells were pyrolyzed at a high temperature forming primarily
condensed aromatic C structures. On one hand, the recalcitrant
nature of biochar may be important if the key goal is to sequester
C in the highly stable SOC pool. On the other hand, if the goals
are to improve soil fertility and also increase C sequestration,
then a biochar having more readily oxidizable structural groups
and a low C:N ratio may be more appropriate. Eventually, the
biochar will oxidize and soil pH will decrease and CEC increase.
The conditions under which feedstocks are pyrolyzed can po-
tentially be designed to produce biochars with single or dual
targeted characteristics either as a C-sequestration amendment,
a soil fertility correction, or both.
ACKNOWLEDGMENTS
This publication is based on work supported by the US
Department of AgricultureYAgricultural Research Service under
the ARS-GRACEnet project.
The authors thank Ms. Sheeneka Green, Mr. Dean Evans,
Dr. Baoshan Xing, and Ms. Mary Kay Amistadi for laboratory
analyses.
* 2009 Lippincott Williams & Wilkins
Copyright @ 2009 by Lippincott Williams & Wilkins. Unauthorized reproduction of this article is prohibited.
Biochar in Coastal Plain Soil
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 Poster Black carbon from rice residues as soil amendment and for carbon sequestration
identificat
ion card 1Haefele SM, 2Konboon Y, 3Knoblauch C, 4Koyama S, 1Gummert M, 1Ladha JK
1 International Rice Research Institute, 2 Ubon Ratchathani Rice Research Center, 3 University of Hamburg, 4 Japan International Cooperation Agency Tsukuba, INTERNATIONAL RICE RESEARCH INSTITUTE
IRRI

1 Background and objectives 3. The effect of carbonized rice husks in greenhouse and field experiments
On highly weathered soils in tropical and subtropical climates, maintenance of soil To investigate the effect of carbonized residue application, we conducted
organic matter is essential to sustain system productivity and avoid rapid soil greenhouse studies and established field experiments in different rice production
degradation. But climatic conditions as well as soil characteristics favor the rapid systems (irrigated and rainfed). Preliminary results indicate beneficial agronomic
decomposition of organic matter. However, several recent studies indicated that effects on poor soils only. Analysis of the effect on soil characteristics has not yet
carbonized plant residues (black carbon), the product of incomplete combustion of been completed.
organic material, could combine characteristics highly beneficial for soil nutrient
Greenhouse experiment
dynamics with high stability against chemical and microbial breakdown. Our ab
a
ab

project investigates i) past and present uses of carbonized rice residues, ii) the b
b
b

agronomic and environmental effects of carbonized rice residues in rice-based

systems, iii) the bio-degradability of carbonized residues in rice soils, and iv) the
possibilities to integrate residue use into the rice production process.

2. Past and current use of carbonized rice residues

Carbonized rice husk are traditionally used in Japan. In and outside of Japan they Siniloan, Philippines Greenhouse experiments were conducted at IRRI and URRRC (NE
Thailand), comparing treatments without, with plain and with
are mainly used in seed beds, in hydroponics and for the cultivation of ornamental carbonized husk application. Fertilizer treatments (none and medium
plants. NPK rate) were superimposed. Four field trials with identical
treatments were established in irrigated systems in the Philippines
Kuntan production in Japan (photo In Japan, carbonized rice husk (Kuntan) has been produced and
provided by A. Kubota) and India (Los Baños, Modipuram), in a rainfed upland system in the
used in agriculture since a long time. Several books on the utilization
Philippines (Siniloan), and in a rainfed lowland system in NE
of Kuntan in agriculture were already published in the 1910’s in
Thailand (Ubon).
Japan. In and outside of Japan, Kuntan was and is used to cover
and protect rice nurseries, as an additive to the culture medium for
ornamental plants, and as a component of growth media for Preliminary results: greenhouse experiments did indicate small
seedlings and hydroponics. It was also reported to function as Los Banos, Philippines biomass and yield increases due to the application of carbonized rice
absorptive for moisture and gas and as water purifier. husks. Even high application rates did not result in increased
leaching of organic carbon. Results of the field trials did not show
Because of the known use and production of carbonized rice husk in
any significant agronomic effect (biomass or grain yield) in irrigated
Japan, the same material and procedure for our experiments was
systems with medium soil fertility (Los Baños, Modipuram), but
Kuntan production at IRRI used. Weight loss due to carbonization was 68%, but the bulk
considerable yield increases were observed in rainfed systems with
density did not change (128 g per liter). Data of the chemical
poor soil fertility. Particularly interesting at these sites and in the
analysis is given below. Note that these results depend highly on the
greenhouse experiments was that the addition of carbonized husks
temperature and duration of the carbonization process.
did increase the fertilizer use efficiency. Analysis of soil related
C N P K Ca Mg Si Fe Zn results is still ongoing.
-----------------------------------g kg-1--------------------------------- ppm
Plain rice husk 362 6.9 1.4 4.5 0.6 0.8 95 0.9 17
Carbonized husk 1 398 19.7 7.8 13.4 1.4 3.7 187 1.2 63 5. Integrated use of rice residues (husks)
Carbonized husk 2 398 4.9 1.1 7.2 1.6 1.2 220 5.6 36
Charcoal (wood) 805 9.4 1.2 11.2 11.8 1.3 1 0.4 21 Husks constitute about 20% of the paddy weight. Using a newly developed rice hull
furnace, they could substitute fossil fuels in the paddy drying process, supply a
4. Bio-degradability of carbonized rice husks in incubation experiments material for soil improvement and contribute to carbon sequestration in the soil.
The decomposition of carbonized organic matter in soils was evaluated under oxic Carbonized rice husk can also be the end product
of rice hull furnaces used by small commercial rice
and anoxic conditions by measuring the carbon dioxide and methane production. mills to dry the paddy. A new semi-automatic
Under both conditions, carbonized rice husks were inert and not decomposed. down-draft rice husk furnace, developed by IRRI,
Hohenheim University (Germany) and Nong Lam
Fig 1: Experimental setup. Anoxic incubations (left panel) were
University (Vietnam) has an adjustable feed
done with field fresh soil in sealed glass flasks. CH4 and CO2-
mechanism for setting the retention time of the
concentrations were measured repeatedly and gas production
rice husk inside the burning chamber.
rates calculated. Oxic carbon turnover was studied in soil
samples incubated in gas tight glass flasks under ambient air
(right panel). A NaOH trap was used to precipitate CO2 At high feed rates the husks are incompletely
produced. After the incubation period (24-48 h) CO2 production burned resulting in a higher ash recovery, i.e. the
was quantified by titration with HCl. furnace produces a high percentage of carbonized
rice husk without loosing much efficiency. Hence,
rice husks could simultaneously substitute fossil
loamy Fluvisol sandy Fluvisol Fig 2: Production of CH4 and CO2 in two temperate Fluvisols. fuels (CO2 emission neutral) during the paddy
Upper panels show CH4 (black) and CO2 (grey) production drying process, supply a material for soil
under anoxic incubation conditions, lower panels CO2- improvement and contribute to carbon
Anoxic

production under oxic conditions. Error bars indicate standard sequestration in the soil.
deviation of quadruplicate incubations, the diamond the
content of Corg in the different treatments. Presented by SM Haefele, IRRI
(shaefele@cgiar.org)

Conclusions:
Oxic

Preliminary results: If dried rice husks were added, the initial Carbonized crop residues could contribute to increase soil
CO2 and CH4 production rates increased by a factor of 3 to 7. carbon storage (carbon sequestration), reduce climate-relevant
In case of the addition of carbonized material, no effect could
No rice
husk
Carboni
zed rice
Untreat
ed rice
No rice
husk
Carboni
zed rice
Untreat
ed rice be measured. These results indicate the strong resistance of gas emissions and improve the natural soil resource in rice-
added husk
added
husk
added
added husk
added
husk
added carbonized organic matter towards microbial breakdown. based production systems.
 Climate Change: Global Risks, Challenges and Decisions IOP Publishing
IOP Conf. Series: Earth and Environmental Science 6 (2009) 372052 doi:10.1088/1755-1307/6/7/372052

P37.45
Biochar as a soil amendment positively interacts with nitrogen fertiliser to improve barley yields in
the UK
Alfred Gathorne-Hardy, J Knight, J Woods
Imperial College, Centre for Energy Policy and Technology (ICEPT), London, UK

Introduction: Soil organic carbon (SOC) is vital for sustainable yields, retaining water and nutrients,
providing a habitat for soil biota and improving soil structure (Lorenz 2007). SOC is also a major carbon
store, containing over twice the total carbon present in the atmosphere. Land Use Change and arable
farming practises have already led to a marked reduction in SOC, and with the increased temperatures
expected with climate change SOC is likely to fall further (Raich, Potter et al. 2002). Its loss reduces soil
fertility and further exacerbates climate change.
Biochar, the use of charcoal as a soil amendment, has been proposed to increase both SOC levels and soil
fertility.
Biochar has two key properties:
1. a high affinity to nutrients and water, reducing onsite nutrient loss and offsite pollution from
nutrient leaching
2. a long residence time. Unlike soil amendments such as compost/manure biochar has a half life of
up to several centuries (Lehmann, Gaunt et al. 2006)

The long residence time has lead to biochar’s promotion for carbon sequestration, as it solves the lack of
permanence problem that plagues most other (non-geological) carbon capture and storage programmes, for
example afforestation. But before biochar can be widely taken up it is essential that its impacts on arable
cropping are understood. To date there has been almost no work looking at the use of biochar in temperate
agriculture.
Aim: To investigate the impacts of different rates of biochar on cereal growth within temperate agriculture,
and specifically the interaction of biochar and nitrogen fertiliser.
Methods: In 2008 a semi-randomised block design was established using spring barley on light land with
five levels of biochar (0, 5 10, 20 and 50 t/ha) and 5 levels of N (ammonium nitrate) fertiliser (0, 25, 50, 70
and 100 kgN/ha). Each biochar level was tested against each N level, giving 25 treatments. Each treatment
was repeated 5 times. Other nutrients were supersaturated on all plots. Results were analysed using
ANOVA in R.
Results and Conclusions: Interestingly the results showed no significant effect of yield for biochar alone, but
do show a significant interaction (p = 0.055) between biochar and N fertiliser. Biochar appears to increase
the nitrogen use efficiency. In this site the addition of 50t/ha of biochar increased the total yield by c. 30%
when high levels of N were used.
A likely explanation for the lack of effect from biochar alone on yield is that one of the most important
attributes of biochar – its ability to retain water – was not tested as the 2008 growing season was
exceptionally wet, so water was unlikely to have been a limiting factor.


c 2009 IOP Publishing Ltd 1
 Climate Change: Global Risks, Challenges and Decisions IOP Publishing
IOP Conf. Series: Earth and Environmental Science 6 (2009) 372052 doi:10.1088/1755-1307/6/7/372052

250
Barley yield (g/250cm2)

200

0 N kg/ha
150
10 N kg/ha
100 20 N kg/ha
80 N kg/ha
50
100 N kg/ha

0
0 5 10 20 50
Biochar application rate (t/ha)

Figure 1. The effect of different levels of biochar, N fertiliser, and their interactions on barley yield

In addition to the onsite experiment, an economic model is being developed to see which price mechanisms,
if any, are required to encourage farmers to use biochar as a regular soil amendment.
Conclusion: These results demonstrate that biochar can have an important role in addressing climate change
through carbon sequestration and increased nitrogen use efficiency, and at the same time improving yields
and food security.
It is hoped that through its high affinity to nutrients and water biochar can help to buffer climatic variability
and reduce the need for fertiliser inputs. Thus biochar could both adapt agriculture to, and mitigate it from,
climate change.

List of References
Lehmann, J., J. Gaunt, et al. (2006). "Bio-Char Sequestration in Terrestrial Ecosystems – a Review." Mitigation and Adaptation
Strategies for Global Change 11: 25.
Lorenz, K. (2007). "Strengthening the soil organic carbon pool by increasing contributions from recalcitrant aliphatic
bio(macro)molecules." Geoderma 142(1-2): 1-10.
Raich, J. W., C. S. Potter, et al. (2002). Interannual variability in global soil respiration, 1980-94. 8: 800-812.

2
African Journal of Agricultural Research Vol. 3 (11), pp. 759-774, November, 2008
Available online at http://www.academicjournals.org/AJAR
ISSN 1991-637X © 2008 Academic Journals

Full Length Research Paper

Preliminary evaluation of the effects of carbonized

chicken manure, refuse derived fuel and K fertilizer
application on the growth, nodulation, yield, N and P
contents of soybean and cowpea in the greenhouse
Seth O. Tagoe, T. Horiuchi and T. Matsui
United Graduate School of Agricultural Science, Gifu University, Yanagido 1-1 Gifu, 501-1193, Japan.
Accepted 3 November, 2008

Carbonized organic materials have not traditionally been applied to grain legumes such as soybean
(Glycine max L. Merrill) and cowpea (Vigna unguiculata L. Walp) although the potential for favourable
agronomic responses exist because of their high contents of available P. We explored the effects of
carbonized chicken manure and carbonized refuse derived fuel (RDF) from municipal organic waste
with or without inorganic K fertilization on the growth, nodulation, seed yield, N and P contents of
soybean and cowpea in a vinyl house pot experiment. Growth, nodulation, plant total N and P contents
were evaluated at peak flowering stage of legume growth. The application of carbonized chicken
manure only increased seed yield by 41 and 146% for soybean and cowpea respectively while the
carbonized chicken manure with inorganic K fertilizer increased seed yield by 53 and 185% for soybean
and cowpea respectively relative to the un-amended control. The application of carbonized RDF only
increased seed yield by 20 and 59% for soybean and cowpea respectively while the application of
carbonized RDF with inorganic K fertilizer increased seed yield by 45 and 126% for soybean and
cowpea respectively relative to the absolute control. The application of both carbonized organic
materials with inorganic K fertilizer increased number of nodules more than their sole application.
Results suggested that the application of carbonized chicken manure and carbonized RDF improved the
growth, nodulation, seed yield, N and P contents of both grain legumes due to their high content of P.
The application of the carbonized organic materials with inorganic K fertilizer further increased seed
yields of both grain legumes suggesting that K was limiting the response to P from the organic
materials in the experimental soil.

Key words: Carbonization, chicken manure, grain legumes, refuse derived fuel, seed yield.

INTRODUCTION

Soybean (Glycine max (L) Merrill) and Cowpea (Vigna
unguiculata (L) Walp) are important grain legumes grown
in the tropics and sub-tropics. Cowpea is particularly
important in West Africa where it occupies 6 million
hectares of agricultural land (Bationo et al., 1990) with
over 9.3 million metric tons of annual production (Ortiz,
1998). The main limiting nutrients for legume production
*Corresponding author. E-mail:sethtagoe@hotmail.com;
Tel: +81-90-1822-9702. Fax: +81-58-293-2846.
in West Africa are N and P (Fox and Kang, 1977). The
high cost and scarcity of inorganic fertilizers had renewed
interest in the use of unorthodox organic soil amendment
materials such as carbonized organic materials (Shinogi
et al., 2003) and bio-char (Ishii and Kadoya, 1994) for
cultivation of crops.
Carbonization has been proposed as a management
tool for agricultural and municipal wastes producing
fertilizer, renewable energy and bio-char. Carbonization is
achieved through pyrolysis of organic wastes at tempera-
o
tures ranging between 300 and 500 C and eliminates the
bad smell, reduces the volume and weight of organic
wastes (Popov et al., 2004). During carbonization, some
760 Afr. J. Agric. Res.
amount of N is lost in the form of ammonia gas but the
resulting carbonized material is higher in available P by
up to 5 times compared to the original waste (Shinogi et
al., 2003; Tagoe et al., 2008).
Carbonized organic wastes can be used as soil
amendments to supply plant nutrients especially N and P.
The application of carbonized organic wastes to soil
improve the physical properties of the soil, improve soil
fertility and nutrient retention (Sombroek et al., 1993;
Lehmann and Rondon., 2005), stimulate microbial
activities in soil (Tejada et al., 2006), increase mycorrhizal
abundance and/or functioning (Warnock et al., 2007),
increase nodule abundance in soybean (Tagoe et al.,
2008) as well as improve biological N2 fixation in common
beans (Rondon et al., 2007).
No work has been reported in the scientific literature on
the use of carbonized refuse derived fuel (CRDF) as soil
amendment for legume growth while only one work had
been reported on the use of carbonized chicken manure
as soil amendment for soybean growth (Tagoe et al.,
2008). In this study, we assess the effects of two
carbonized organic soil amendment materials (chicken
manure and RDF) on the growth, nodulation, yield, N and
P concentrations of two grain legumes (soybean and
cowpea) with or without inorganic K fertilizer. We
hypothesized that the application of carbonized organic
materials will improve the growth, nodulation, seed yield,
N and P concentrations of grain legumes more with K
fertilizer than without K fertilizer because K could limit the
response of grain legumes to P from carbonized organic
materials in low P status soils (Carsky, 2003) and also
because of the low K content of the experimental soil.
Our major objectives are to elucidate the effect of
carbonized chicken manure and carbonized RDF on 1)
the growth, nodulation and seed yield and 2) plant and
seed concentrations of N and P of both crops.
MATERIALS AND METHODS
Plant culture
Experiments were conducted from May to October, 2007 at the
greenhouse of Gifu University Experiment Farm (35o 27’ N, 136o 46’
E). Wagner pots (1/2000a) were filled with sandy loam soil with the
following characteristics: pH; 5.62, EC; 0.28 mS cm-1, total N;
0.100%, total C; 0.88%, available P; 1.16 mg 100 g-1, available K;
13.3 mg 100 g-1, available Ca; 90.0 mg 100 g-1, and available Mg;
31.0 mg 100 g-1. The nutrient concentrations, pH and EC of the
carbonized chicken manure and RDF used in the experiment are
shown in Table 1. Carbonized chicken manure was obtained from
Tokyo Yougyou Kabushiki Kaisha, Tajimi City, Gifu Prefecture,
Japan. Carbonized chicken manure was produced from pelleted,
dried chicken manure through pyrolysis at a temperature of 450oC
for one hour in a furnace. Carbonized RDF was obtained from
Kurimoto Tekkosho, In a City, Gifu Prefecture, Japan. Carbonized
RDF was prepared from municipal organic waste through pyrolysis
at a temperature of 500oC for 2 h in a kiln after drying the waste in a
furnace for 10 h. The amounts of the carbonized organic materials
applied per pot for the various treatments are shown in Table 2.
Carbonized organic amendments were applied to the appropriate
treatments three weeks before sowing. Potassium fertilizer was
applied as Muriate of Potash (KCl) at a rate of 83 kg K ha-1 to the
appropriate treatments at sowing. No chemical pesticides were
used in this experiment. Five seeds of soybean (Glycine max (L)
Merrill cv Akishirome) and cowpea (V. unguiculata (L) Walp cv
Tsurushi sasage) were sown per pot on 7th June, 2007. After
emergence, seedlings were thinned to two per pot. Plants were
watered as when necessary. All pots were kept completely free of
weeds within the duration of the experiment by hand-picking when
they appear.
Experimental design
The experiment was set up using two grain legumes (soybean and
cowpea), two carbonized organic materials (carbonized chicken
manure and carbonized RDF) with two rates of inorganic K fertilizer
with or without K, 83 kg K ha-1 and 0 kg K ha-1 respectively in a
factorial combination giving a total of 48 treatments (2 x 2 x 2 x 6)
arranged in completely randomized design (CRD) of six
replications. All data collected from the study were analyzed by
using Duncan’s Multiple Range Test (DMRT) (Excel Statistical
Package Version 6.0) and mean separations were done by the
same method.
Measurements
Plant height and relative chlorophyll content (SPAD) were
measured twice within the duration of the experiment. For both
parameters, the first (pre-flowering) and second (post-flowering)
samplings were done on 16th July and 31st July respectively. SPAD
was measured with a chlorophyll meter model SPAD-502 (Minolta
Co. Ltd., Japan). SPAD readings were taken from 12 randomly
selected youngest and fully expanded leaves of plants in each pot.
Each SPAD reading was taken on one side of the mid-rib of the leaf
blade, midway between the leaf blade and tip.
Sampling for leaf area, dry matter weight, and number of nodules
was done on 26th July for cowpea and 1st August for soybean
when the plants were at peak flowering stage. Shoots were
harvested by cutting the plants in each pot at the soil level. Roots
were harvested by lifting the soil in each pot and washing off the
soil under running water from a tap. The harvested shoots were
separated into leaves, stems (including petioles), flowers etc. The
roots were washed clean of soil and root nodules were separated
and counted. The leaves were used to estimate leaf area by the
core borer method after oven-drying harvested samples at 80oC for
72 h. The dried samples were then weighed to determine dry
weights of leaves, stems, roots and nodules and finally total dry
matter weight.
The total dry material was milled to pass 0.5 mm mesh sieve
using a wonder blender (Model WB-1). The milled plant samples
were used for total N and total P analyses and determination. Total
N was determined with an automatic high sensitive NC analyzer
(Sumigraph NC-95A, Shimadzu Co. Ltd., Japan). Total P was
determined colorimetrically (HITACHI-U-1800) according to Bray
and Kurtz (1945) and Murphy and Riley (1962).
Residual experimental soil exchangeable cations (K+, Ca2+, and
Mg2+) were measured with a Polarized Zeeman Atomic Absorption
Spectrophotometer (HITACHI-180-60) after extraction of samples
with 1.0N Ammonium acetate solution (pH 7.0).
RESULTS
Plant growth
At the pre-flowering sampling of the relative chlorophyll
 Tagoe et al. 761

Table 1. Nutrient composition of carbonized organic materials used in the experiment after pyrolysis.

Property Carbonized chicken Manure Carbonized refuse derived fuel

pH † 9.93 7.76
EC (µS cm-1) 3.64 65.03
Total N (g kg-1) 44,0 20.2
NO3-N (mg kg-1) 1.90 -
Total C (g kg-1) 497.5 500.9
C/N ratio 11.31 25.00
Na (g kg-1) 2.37 3.38
P (g kg-1) 24.6 5.22
K (g kg-1) 37.00 3.45
Ca (g kg-1) 7.67 30.28
Mg (g kg-1) 1.93 2.10

† The pH and EC were measured in the extracts of carbonized organic materials in distilled water
[ 1:20 (w/v)] on dry weight basis.

Table 2. Treatment details showing amounts of carbonized organic materials and inorganic K fertilizer applied per pot and treatment
abbreviations.

Treatment Amount of carbonized Amount of K fertilizer Abbreviation

organic material applied as muriate of
applied (g) potash (KCl) (mg)
No carbonized organic material (Control) 0 0 Control (Without K)
No carbonized organic material (Control) 0 830.0 Control (With K)
Carbonized chicken manure 11.40 0 CCM (Without K)
Carbonized chicken manure 11.40 410.0 CCM (With K)
Carbonized refuse derived fuel 24.75 0 CRDF (Without K)
Carbonized refuse derived fuel 24.75 740.0 CRDF (With K)

content (SPAD) leaves of soybean plants treated with
carbonized chicken manure were the greenest. Leaves of
soybean plants treated with carbonized RDF were of
intermediate greenness while those of the control were
the least green with or without K fertilizer. Generally,
leaves of soybean plants without K fertilizer were greener
than those with K fertilizer (Figure 1). The observed
differences were significant according to DMRT at p<
0.05.
At the post-flowering sampling of SPAD, a similar trend
was observed for the control and carbonized RDF
treatments with or without K fertilizer. However, leaves of
soybean plants treated with carbonized chicken manure
were the greenest with or without K fertilizer (Figure 1). At
the pre-flowering sampling of SPAD of cowpea, leaves of
plants treated with carbonized chicken manure with or
without K fertilizer were the greenest. Cowpea leaves of
plants treated with carbonized RDF with or without K
fertilizer were of intermediate greenness. Leaves of
cowpea plants without any organic amendment (control)
with or without K fertilizer were least green (Figure 2). For
cowpea leaves of plants treated with carbonized chicken
manure and carbonized RDF, no difference in greenness
were observed for treatments with and without K fertilizer.
Leaves of control cowpea plants that received K fertilizer
were greener than those that did not (Figure 2). At the
post-flowering sampling of SPAD for cowpea, a similar
trend to the pre-flowering SPAD was observed except
that leaves of control cowpea plants without K fertilizer
were greener than those with K fertilizer (Figure 2). The
observed differences among treatments for both pre-
flowering and post-flowering were significant at p<0.05
according to DMRT.
Total dry weight was significantly highest according to
DMRT at p<0.05 in soybean plants treated with
carbonized chicken manure followed by those treated
with carbonized RDF. Control soybean plants were least
heavy in total dry weight (Table 3). For all treatments,
soybean plants with K fertilization produced heavier dry
matter than the corresponding treatments without K
fertilization (Table 3). There was a strong and significant
positive relationship between total dry weight and plant
2
total N content of soybean (R = 0.79 **, Figure 3a). A
strong and significant positive relationship was observed
between soybean total dry weight and plant total P
2
content (R = 0.90 **, Figure 3b).
Total dry weight of cowpea followed a similar trend as
that of soybean. Cowpea plants treated with carbonized
762 Afr. J. Agric. Res.

Figure 1. Effects of carbonized organic materials supply on the pre-flowering and post-flowering
SPAD values of soybean. Data show the means ± SD of three independent replications.

Figure 2. Effects of carbonized organic materials supply on the pre-flowering and post-flowering
SPAD values of cowpea. Data show the means ± SD of three independent replications.

chicken manure produced significantly the highest dry produced the least dry matter (Table 4). For all
matter followed by those treated with carbonized RDF treatments, cowpea plants that received K fertilizer
according to DMRT at p<0.05. Control cowpea plants produced heavier dry matter than those that did not
 Tagoe et al. 763

Table 3. Growth, nodulation, total N and P contents of soybean as affected by the application of carbonized organic materials.

Treatment K fertilizer Total dry Number of Plant total N Seed total N Plant total P Seed total P
weight (g) nodules content (g content (g content (g content (g
kg-1) kg-1) kg-1) kg-1)
Control Without K 38.0 a 380.7 a 27.8 a 66.1 a 0.35 a 0.09 a
With K 49.3 b 428.3 b 27.2 a 66.8 a 0.38 b 0.11 b
CCM Without K 64.2 c 529.3 c 31.9 c 72.8 d 0.44 d 0.27 c
With K 81.8 d 611.0 d 33.0 d 73.8 e 0.47 f 0.34 e
CRDF Without K 54.1 b 437.3 b 30.4 b 70.8 b 0.43 c 0.26 c
With K 69.2 c 567.0 c 30.8 b 71.8 c 0.45 e 0.31 d
Within each column, means having a common letter(s) are not significantly different according to DMRT at p < 0.05.

Table 4. Total dry weight, number of nodules, total N and P contents of cowpea as affected by the application of carbonized
organic materials.

Treatment K fertilizer Total dry Number of Plant total Seed total Plant total Seed total
weight (g) nodules N content N content P content P content
(g kg-1) (g kg-1) (g kg-1) (g kg-1)
Control Without K 17.5 a 103.3 a 23.1 a 40.3 a 0.54 a 0.12 a
With K 31.7 bc 140.0 b 22.9 a 41.3 b 0.56 b 0.14 b
CCM Without K 34.8 c 122.0 ab 27.3 c 46.6 e 0.64 d 0.18 d
With K 38.3 d 215.0 d 31.9 d 47.9 f 0.67 e 0.21 f
CRDF Without K 29.3 b 113.0 a 24.9 b 43.5 c 0.62 c 0.16 c
With K 37.7 d 189.0 c 28.0 c 44.9 d 0.64 d 0.19 e
Within each column, means having a common letter(s) are not significantly different according to DMRT at p < 0.05.

(Table 4).There was a significant positive relationship
between total dry weight and plant total N content of
2 2
cowpea (R = 0.56 *, Figure 4a). A strong and significant
positive relationship was observed between total dry
2 2
weight and plant total P content of cowpea (R = 0.70 **,
Figure 4b).
Number of nodules of soybean was significantly highest
in soybean plants treated with carbonized chicken
manure followed by those treated with carbonized RDF
according to DMRT at p<0.05. Control soybean plants
produced the least number of nodules. For each
carbonized organic amendment treatment, soybean
plants that received K fertilizer produced more nodules
than those that did not (Table 3). There was a strong and
significant positive relationship between number of
2
nodules and plant total N content of soybean (R =
0.75**, Figure 5a). Also, a strong and significant positive
relationship was observed between number of nodules
2
and plant total P content of soybean (R = 0.83**, Figure
5b).
Cowpea plants of both carbonized organic amendment
treatments without K fertilization produced similar number
of nodules. However, cowpea plants treated with
carbonized chicken manure and K fertilizer produced
significantly the greatest number of nodules followed by
cowpea plants treated with carbonized RDF and K
fertilizer according to DMRT at p<0.05. Control cowpea
plants with K fertilizer produced the least number of
nodules (Table 4). There was a significant positive
relationship between number of nodules and plant total N
content of cowpea (R = 0.68 *, Figure 6a). No significant
positive relationship was observed between number of
nodules and plant total P content of cowpea (R = 0.43
ns, Figure 6b).
Yield and yield components
Dry pod yield and seed yield of soybean followed a
similar trend. Soybean seed yield was heaviest in
treatments amended with carbonized chicken manure
with or without K fertilizer followed by carbonized RDF
amended treatments with or without K fertilizer. Soybean
seed yield was least in control treatments with or without
K fertilizer (Table 5). Generally, carbonized organic
amendment treatments that received K fertilizer produced
better dry pod and seed yields than those without K
fertilizer. The application of carbonized chicken manure
only increased soybean seed yield by 41% while the
application of carbonized RDF only increased soybean
seed yield by 20%. The application of carbonized chicken
manure and K fertilizer increased soybean seed yield by
53% while the application of carbonized RDF and K
fertilizer increased soybean seed yield by 45%.
Dry pod yield and seed yield of cowpea followed a
similar trend. Cowpea plants treated with carbonized
764 Afr. J. Agric. Res.

= Without K fertilizer; = with K fertilizer; ** = significant at 1%.

= Without K fertilizer; = with K fertilizer; ** = significant at 1%.

Figure 3. Relationship between total dry weight, plant total N content and plant total P content of soybean
at peak flowering.
3a. Relationship between total dry weight and plant total N content of soybean at peak flowering stag
3b. Relationship between total dry weight and plant total P content of soybean at peak flowering stage
 Tagoe et al. 765

= Without K fertilizer; = with K fertilizer; ** = significant at 1%.

= Without K fertilizer; = with K fertilizer; * = significant at 5%.

Figure 4. Relationship between total dry weight, plant total N content and plant total P content of cowpea
at peak flowering.
4a. Relationship between total dry weight and plant total N content of cowpea at peak flowering stage.
4b. Relationship between total dry weight and plant total P content of cowpea at peak flowering stage.
766 Afr. J. Agric. Res.

= Without K fertilizer; = with K fertilizer; ** = significant at 1%.

= Without K fertilizer; = with K fertilizer; ** = significant at 1%.

Figure 5. Relationship between number of nodules, plant total N content and plant total P content of soybean at
peak flowering.
5a. Relationship between number of nodules and plant total N content of soybean at peak flowering stage.
5b. Relationship between number of nodules and plant total P content of soybean at peak flowering stage.
 Tagoe et al. 767

= Without K fertilizer; = with K fertilizer; = significant at 5% .

(6a)

= Without K fertilizer; = with K fertilizer; ns = not significant .

6b)
Figure 6. Relationship between number of nodules, plant total N content and plant total P content of cowpea at peak
flowering.
6a. Relationship between number of nodules and plant total N content of cowpea at peak flowering stage.
6b. Relationship between number of nodules and plant total P content of cowpea at peak flowering stage.
768 Afr. J. Agric. Res.

Table 5. Yield and yield components of soybean as affected by the application of carbonized organic materials.

Treatment K fertilizer Dry pod Seed yield No. of No. of 100 seed Harvest
yield (g) (g) pods seeds wt (g) index
plant-1 pod-1
Control Without K 49.5 a 38.1 a 50.4 a 1.8 a 26.4 a 0.54 ab
With K 68.7 c 52.9 c 108.7 c 1.8 a 28.0 b 0.63 e
CCM Without K 69.8 c 53.7 c 79.0 c 1.9 b 28.9 c 0.57 bc
With K 71.9 cd 58.4 d 137.5 d 2.0 c 29.9 e 0.59 bc
CRDF Without K 59.7 b 45.9 b 68.9 b 1.9 b 27.8 b 0.67 e
With K 75.8 d 55.4 cd 103.7 d 1.9 b 29.6 d 0.52 a

Table 6. Yield and yield components of cowpea as affected by the application of carbonized organic materials.

Treatment K fertilizer Dry pod Seed yield No. of pods No. of 100 seed Harvest
yield (g) (g) plant-1 seeds pod-1 wt (g) index
Control Without K 26.1 a 14.2 a 9.4 a 11.7 ab 17.4 a 0.66 d
With K 40.6 c 27.6 c 12.0 bc 11.3 ab 17.3 a 0.60 ab
CCM Without K 47,9 d 34.9 e 11.9 b 12.0 b 19.8 b 0.61 bc
With K 54.9 e 40.5 f 14.9 d 12.3 b 20.2 b 0.63 c
CRDF Without K 35.6 b 22.6 b 11.4 b 10.7 a 18.1 a 0.59 ab
With K 47.1 d 32.1 d 13.0 c 11.3 ab 19.6 b 0.58 a

chicken manure with or without K fertilizer produced the
heaviest seed yield followed by those treated with
carbonized RDF. Control cowpea plants with or without K
fertilizer produced the least heavy seed yield. Generally,
cowpea plants that received K fertilizer produced heavier
seed yield than those that did not receive K fertilizer
(Table 6). Cowpea seed yield increased 146% by the
application of carbonized chicken manure only while it
increased by 49% as a result of the application of
carbonized RDF only. The application of carbonized
chicken manure and K fertilizer increased cowpea seed
yield by 185% while the application of carbonized RDF
and K fertilizer increased it by 126%.
Number of pods/plant of soybean followed a similar
pattern to dry pod and seed yields of soybean. In all
treatments, number of pods/plant of soybean was better
for treatments that received K fertilizer than for those
without K fertilizer (Table 5).
Number of pods/plant was highest in cowpea plants that
received K fertilizer than those that did not for all
treatments. Number of pods/plant was significantly
highest in cowpea plants treated with carbonized chicken
manure followed by carbonized RDF. Number of
pods/plant was least in control cowpea plants.
Number of seeds per pod was only slightly increased by
K fertilizer in soybean plants treated with carbonized
chicken manure (Table 5).
Number of seeds/pod of cowpea was neither affected
by carbonized organic amendment material supply nor K
fertilizer application (Table 6).
100 seed weight of soybean was increased by K fertilizer
in all organic amendment treatments. 100 seed weight
was significantly heaviest in carbonized chicken manure
treated plants according to DMRT at p< 0.05 followed by
carbonized RDF treated plants. 100 seed weight was
least heavy in control soybean plants (Table 5).
100 seed weight of cowpea was not affected by K
fertilizer application in control and carbonized chicken
manure treated plants but was increased slightly in
carbonized RDF treated plants. 100 seed weight of
cowpea was highest in plants treated with carbonized
chicken manure with or without K fertilizer and similar to
plants treated with carbonized RDF and K fertilizer (Table
6).
Harvest index was increased by K fertilizer application
in control soybean plants and reduced in carbonized RDF
treated soybean plants. K fertilizer application did not
affect harvest index in carbonized chicken manure
treated soybean plants (Table 5).
Harvest index of cowpea did not follow any particular
trend but was highest in control cowpea plants without K
fertilizer and lowest in carbonized RDF treated plants with
K fertilizer (Table 6).
N contents
Plant total N content of soybean was highest in
carbonized chicken manure amended plants followed by
carbonized RDF amended plants. Control soybean plants
were least in plant total N content (Table 3). K fertilizer

only slightly increased plant total N content of carbonized
chicken manure amended soybean plants while it had no
effect on plant total N content of carbonized RDF
amended soybean plants (Table 3). Seed total N content
of soybean followed a similar trend as plant total N
content of soybean. The only difference is that K fertilizer
application increased seed total N content of carbonized
chicken manure and carbonized RDF amended soybean
plants while it had no effect on seed total N content of
control soybean plants (Table 3). There was a significant
positive relationship between seed total N content and
2
seed yield of soybean (R = 0.50*, Figure 7).
Plant total N content of cowpea was highest in plants
treated with carbonized chicken manure with or without K
fertilizer followed by plants treated with carbonized RDF
with or without K fertilizer. Control cowpea plants were
lowest in plant total N content with or without K fertilizer
(Table 4). K fertilizer application increased plant total N
content in carbonized chicken manure and carbonized
RDF amended cowpea plants but not in control cowpea
plants (Table 4). Seed total N content of cowpea followed
a similar trend (Table 4). However, K fertilizer application
increased cowpea seed total N content in all treatments
(Table 4). There was a strong positive relationship
between plant total N content and seed yield of cowpea
2
(R = 0.72**) as well as between seed total N content and
2
seed yield of cowpea (R = 0.81**, Figure 8).
P contents
Plant total P content of soybean was affected by both
carbonized organic amendment supply and K fertilizer
application. Plant total P content of soybean was highest
in plants amended with carbonized chicken manure with
or without K fertilizer followed by carbonized RDF treated
plants with or without K fertilizer. Control soybean plants
were lowest in plant total P content (Table 3). K fertilizer
application increased plant total P contents of all
treatments (Table 3). Seed total P content of soybean
followed a similar trend as plant total P content of
soybean (Table 3). There was a positive relationship
between plant total P content and seed yield of soybean
2
(R = 0.62*, Figure 9).
Plant total P content of cowpea was affected by both
carbonized organic amendment and K fertilizer
application. Plant total P content of cowpea was highest
in carbonized chicken manure amended cowpea plants
with or without K fertilizer followed by carbonized RDF
amended plants with or without K fertilizer. Plant total P
content was lowest in control cowpea plants (Table 4). K
fertilizer application increased plant total P contents of
cowpea in all treatments. Seed total P content of cowpea
followed a similar trend as plant total P content of cowpea
(Table 4). There was a strong positive relationship
between plant total P content and seed yield of cowpea
2
(R = 0.71**, Figure 10). There was a strong positive
relationship between seed total P content and seed yield
Tagoe et al. 769
2
of cowpea (R = 0.83**, Figure 10).
DISCUSSION
The effects of two carbonized organic materials that is,
chicken manure and refuse derived fuel (RDF) with or
without inorganic K fertilizer on the growth, nodulation,
yield, N and P contents of two grain legumes i.e. soybean
and cowpea were explored in this greenhouse trial. The
application of carbonized chicken manure only increased
seed yield by 41 and 146% in soybean and cowpea
respectively while the application of carbonized chicken
manure and inorganic K fertilizer increased seed yield by
53 and 185% in soybean and cowpea respectively. The
application of carbonized RDF only increased seed yield
by 20 and 59% in soybean and cowpea respectively
while the application of carbonized RDF and inorganic K
fertilizer increased seed yield by 45 and 126% in soybean
and cowpea respectively. The trends for dry pod yield and
total dry weight of both grain legumes were similar to that
of their respective seed yields. This result is consistent
with that reported by Rondon et al. (2007) who observed
a 46% increase in common bean yield over the control in
response to bio-char application. Chan et al. (2007)
observed in their work that in the absence of N fertilizer,
green-waste bio-char application to soil did not increase
radish yield even at higher rates but reported significant
bio-char and N fertilizer interaction highlighting the role of
bio-char in improving N fertilizer use efficiency of the
plant. Although few works are available for comparison
with the results of this study, our results are consistent
with previous works done using ordinary un-carbonized
chicken manure. Garcia and Blancaver (1983) reported
that the application of poultry manure increased soybean
seed yield by 62% over the control. Schmidt et al. (2001)
found that soybean seed yield increased linearly with
increasing swine manure rate. Several researchers have
reported soybean seed yield increases with applied
commercial N fertilizer (Lamb et al., 1990; Wesley et al.,
1998). The higher seed yields of both grain legumes in
response to the application of carbonized organic
materials especially carbonized chicken manure suggest
that both N and P are important nutrients that influence
the growth and yield of soybean and cowpea. Carbonized
chicken manure and carbonized RDF contain high levels
of macro-nutrients especially N and P as well as micro-
nutrients that would be available to plant roots and soil
biota. Hariston et al. (1990) and Schmidt et al. (2001)
observed that soybean not only requires a considerable
amount of N to produce a crop, but also a constant
supply of available P to maintain rapid growth and
development. Carbonized chicken manure and
carbonized RDF are particularly rich in both total and
available P and their application to soil at high rates can
supply a considerable amount of available P for plant
uptake. Again, carbonized organic materials such as
770 Afr. J. Agric. Res.

= Without K fertilizer; = with K fertilizer; = significant at 5%.

Figure 7. Relationship between seed total N content and seed yield of soybean.

Figure 8. Relationship between plant total N content, seed total N content and seed yield of cowpea.

carbonized chicken manure, carbonized RDF and bio- retaining nutrients and improving soil physical and
char can act as soil conditioners to enhance plant growth biological properties (Glaser et al., 2002; Lehmann and
by supplying macro and micro-nutrient elements, Randon, 2005). Abdelhamid et al. (2004) found increased
 Tagoe et al. 771

= Without K fertilizer; = with K fertilizer; * = significant at 5%.

Figure 9. Relationship between plant total P content at peak flowering and seed yield of soybean.

Figure 10. Relationship between plant total P content, seed total P content and seed yield of cowpea.
772 Afr. J. Agric. Res.
dry weight of faba bean as a result of chicken manure
supply. Cowpea responded positively to the application of
carbonized chicken manure and CRDF in this study in
terms of growth parameters and seed yield. This
response underscores the importance of P as a critical
nutrient element influencing the performance of cowpea
and soybean. Several researchers have reported
significant responses of cowpea and soybean to P
application (Tenebe et al., 1995; Ankomah et al., 1995;
Okeleye and Okelana, 1997). They observed significant
increases in total dry matter yield, number of flowers, pod
and seeds per plant, seed yield and nodulation as a
result of P application. Rondon et al. (2007) reported a
39% increase in biomass production of common bean in
response to bio-char application which they attributed to
greater availability of K, Ca and P. Chan et al. (2007)
observed additional increase in dry matter of radish in the
presence of N fertilizer varied from 95% in the control to
266% in the 100 t/ha bio-char amended soils. Carsky
(2003) working on the response of cowpea and soybean
to P and K on terre de barre soils in southern Benin,
reported that soybean and cowpea grain yield increased
by 147% and 95% respectively in response to P
application.
Number of nodules of soybean and cowpea increased
with the application of carbonized organic materials. This
observation is consistent with that of Tagoe et al. (2008)
who reported increased nodule abundance of soybean as
a result of carbonized chicken manure application and
attributed it to the high available P content of carbonized
chicken manure. The application of carbonized chicken
manure increased nodule abundance of both grain
legumes more than carbonized RDF. This observation
can be attributed to the high P content of carbonized
chicken manure relative to carbonized RDF. Rondon et
al. (2007) also observed that biological N2 fixation of
common bean increased with bio-char application. They
reported that the proportion of fixed N increased from
50% without bio-char addition to 72% with 90 g kg-1 bio-
char added and attributed this observation to greater
availability of B, and Mo and to a lesser extent K, Ca and
P availability. The application of the carbonized organic
materials plus K fertilizer further increased the number of
nodules of both legumes thus confirming the hypothesis
that K could limit the response of P from the carbonized
organic materials in low P status soils (Carsky, 2003). P
fertilization has been shown to increase number of
nodules and their weight in soybean (Cassman et al.,
1993; Jones et al., 1977) and in cowpea (Wan Othman et
al., 1991). This is because P is known to initiate nodule
formation, increases the number of nodule primordia and
is essential for the development and functioning of
formed nodules (Waluyo et al., 2004). Nodules are known
to be a strong sink for P and P concentration in nodules
can be three times higher than in other plant organs with
a minimum effect from P deficiency (Vadez et al., 1999).
Several researchers have reported that the supply of P
plays important roles in establishment, growth and
function of nodules (Israel, 1987; Beck and Munns, 1984;
Leung and Bottomley, 1987) and growth of host plants
(Munns et al., 1981). Number of nodules is important
because a positive correlation has been reported
between nodule number and total nitrogenase activity of
soybean (Singleton and Bohlool, 1984) and alfalfa
(Porter, 1983).
According to Gates and Muller (1979) the application of
fertilizer containing N, P and S to soybean contributed to
forming a stronger symbiotic mechanism and more active
N2-fixation. Since the carbonized organic materials used
in this experiment especially carbonized chicken manure
contain N, P and S (Sharpley et al., 1993) their
application to soybean and cowpea could be beneficial to
the symbiotic N2-fixing mechanism. Low P availability is
especially problematic for leguminous crops because
legume nodules responsible for N2-fixation have high P
requirement (Vance, 2001). P is essential for plant
growth, nodulation and N2-fixation (Pereira and Bliss,
1989). Nodule number as well as nodule dry weight is
greatly reduced by P deficiency (Ribet and Drevon, 1995)
and nitrogenase activity varies with P availability (Israel,
1987). Acute P deficiency is known to prevent nodulation
of grain legumes. P deficiency is more likely to affect N2-
fixation legumes than other species because symbiotic
N2-fixation is an energetically expensive process which
requires more P than does plant growth (Olivera et al.,
2004). The strong positive relationship between number
of nodules and plant total P content of soybean confirms
the importance of P in legume nodulation.
Total N and total P contents of plant and seed of
soybean and cowpea increased with the application of
carbonized organic materials with or without inorganic K
fertilizer. This observation is consistent with that reported
by Jassen (1998) that nutrient uptake requires N, P and K
in balance to reach maximum values. Adeli (2005)
observed increased N concentration in the above ground
biomass of soybean in response to poultry manure
application. The application of chicken manure to faba
bean increased total N contents in roots, shoots and the
whole plant (Abdelhamid et al., 2004). Belle (2006)
reported significant increases in the uptake of N and P in
common bean as a result of the application of N and P
fertilizers. Barker and Sawyer (2005) observed that N
concentration in plant dry matter of soybean was
increased significantly with applied N. Studies with
several legumes have consistently shown a positive
response to P application; whole plant N concentration
and plant dry matter were found to increase in response
to phosphate in the growth media (Pereira and Bliss,
1987). P application has been reported to influence the
contents of other nutrients in cowpea leaves (Kang and
Nangju, 1983), shoots (Bagayoko et al., 2000) and seed
(Omueti and Oyenuga, 1970). Seed total N content of
soybean was positively correlated with seed yield. Also
plant total N and seed total N contents of cowpea were

correlated with seed yield of cowpea. These observations
suggest the importance of N in the seed yields of these
grain legumes due to the high protein contents of their
seeds especially soybean. Plant total P content of
soybean was positively correlated with soybean seed
yield. Also, plant total P and seed total P contents were
positively correlated with seed yield in cowpea. These
observations underscore the importance of not only N,
but P as well in increasing the growth and seed yield of
these grain legumes.
Bio-char addition can result in elevated quantities of
bio-available nutrients such as N, P and metal ions, in the
soil (Tryon, 1948; Lehmann et al., 2003; Gundale and
DeLuca, 2006; DeLuca et al., 2006). Addition of bio-char
to soil alters important soil physical and chemical
properties such as pH (Lucas and Davis, 1961) and
typically increase soil cation exchange capacity (CEC)
(Glaser et al., 2002), and can lead to greater water
holding capacity (WHC) while generally decreasing bulk
density (Tryon, 1948) as well as increase bio-available P
and cations in soils. Improvement in physical properties
of the soil as a result of the addition of carbonized
chicken manure and carbonized RDF to a lesser extent,
may have contributed to the increased growth and yield
of the grain legumes.
Conclusion
The growth, nodulation, seed yield, total N and P
contents of plant and seed of soybean and cowpea all
increased with the supply of carbonized chicken manure
and carbonized RDF due to their high available P
contents. The application of carbonized organic materials
and inorganic K fertilizer increased grain yield and other
parameters in both crops more than the sole application
of the carbonized organic materials thus suggesting that
it is a good potential option to be evaluated for use in the
field and home garden for grain legume production.
ACKNOWLEDGEMENT
The authors would like to express their profound gratitude
to the Ministry of Education, Culture, Sports, Science and
Technology, Japan for financial support without which this
research would not have been possible.
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Soil Science and Plant Nutrition (2006) 52, 489–495 doi: 10.1111/j.1747-0765.2006.00065.x

ORIGINAL ARTICLE
Effect
M. Yamato
of charcoal
Blackwell et al. application
Publishing, Ltd. on crop yield

Effects of the application of charred bark of Acacia mangium on

the yield of maize, cowpea and peanut, and soil chemical
properties in South Sumatra, Indonesia
Masahide YAMATO1, Yasuyuki OKIMORI1, Irhas Fredy WIBOWO2,
Saifuddin ANSHORI2 and Makoto OGAWA3
1
Biological Environment Institute, Research and Development Department, The General Environmental Technos Company,
Kyoto 611-0021, 3Osaka Institute of Technology, Osaka 535-8585, Japan; and 2Research and Development, PT Musi Hutan
Persada, Jl. Residen H. Abdul Rozak 99, Palembang 30114 South Sumatra, Indonesia

Abstract
Charred bark of Acacia mangium (bark charcoal), which is made of wood waste from pulp production,
was applied as soil amendment for the cultivation of maize, cowpea and peanut to examine its effects on
crop yield and soil chemical properties in South Sumatra, Indonesia. The yields of maize and peanut signifi-
cantly increased after the application of bark charcoal under a fertilized condition in an infertile soil
environment. In addition, increases in the root amount and colonization rate of arbuscular mycorrhizal
(AM) fungi after bark charcoal application were also observed in maize. In general, the application of bark
charcoal induced changes in soil chemical properties by increasing the pH value, total N and available P2O5
contents, cation exchange capacity, amounts of exchangeable cations and base saturation, and by decreas-
ing the content of exchangeable Al3+. The amelioration of the soil chemical properties could be effective in
highly weathered infertile tropical soils. The application of charcoal in agriculture is expected to lead to
the formation of a carbon sink in soil and to increase crop yield because it has been suggested that charcoal
is highly resistant to abiotic and biotic degradation, even in a soil environment.
Key words: Arachis hypogaea L., arbuscular mycorrhizal fungi, bark charcoal, Vigna unguiculata L., Zea mays L.

INTRODUCTION cation exchange capacity (CEC), and amelioration of

It is widely recognized that charcoal can be used as soil
amendment in agriculture (Ogawa 1994). The effect of
charcoal application on yield increases in soybean, pea,
moong (Iswaran et al. 1980), sugarcane (Uddin et al.
1994, 1995) and sweet potato (Islam et al. 2000) has
been reported. Igarashi (2002) reported that rice husk
charcoal application increased the yield of maize, soy-
bean and peanut in Indonesia. Glaser et al. (2002) pointed
out that the effects of charcoal can be attributed to
increases in soil pH and the contents of available nutri-
ents, improvement of nutrient retention by increased
Correspondence: M. YAMATO, Biological Environment
Institute, Research and Development Department, The General
Environmental Technos Company, 8-4 Ujimatafuri, Uji, Kyoto
611-0021, Japan. Email: yamato_masahide@kanso.co.jp
Received 3 April 2006.
Accepted for publication 14 March 2006.
soil physical properties such as soil water retention and
aggregation.
In South-East Asia, industrial plantations of the fast-
growing tree, Acacia mangium, have been promoted for
pulp production, and a large amount of wood waste,
mainly bark, is being discharged from pulp mills.
To effectively use the wood waste, Ogawa (1997)
proposed a scheme to utilize charred wood waste as soil
amendment in agriculture. Ogawa (1997) designated
this scheme the “Carbon Sequestration by Forestation
and Carbonization (CFC)” because a carbon sink could
be formed in the soil and the wood waste could be used
effectively. The feasibility of CFC has been examined by
Okimori et al. (2003) in cooperation with PT Musi
Hutan Persada (MHP), an industrial plantation com-
pany, and PT Tanjungenim Lestari Pulp and Paper
(TELPP), a pulp production company, in South
Sumatra, Indonesia. To implement the CFC project, it
would be necessary to promote the utilization of bark

© 2006 Japanese Society of Soil Science and Plant Nutrition

490 M. Yamato et al.
charcoal. Although bark composts have been used as
soil amendment in agriculture and horticulture, there
are few studies examining the application of bark
charcoal.
The purpose of the present study was to investigate
the effects of the application of bark charcoal of
A. mangium on the yield of maize, cowpea and peanut
and its effect on the soil chemical properties. For maize,
the effects on the root dry weight and colonization
rate of arbuscular mycorrhizal (AM) fungi were also
examined. In addition, the role of charcoal application
in the formation of a carbon sink was considered.
MATERIALS AND METHODS
Experimental site
For the experiment examining the application of bark
charcoal and agricultural crops, three types of farm-
land, Sites A, B and C, were selected near the boundary
of the tree plantation of MHP. Site A was located in a
garden of a farmhouse. Site B was located in a garden
reclaimed from a chicken farm. Site C was located in a
farmland reclaimed from grassland. All the sites had
been newly reclaimed for the current experiment and no
fertilizers had been applied previously.
Charcoal
Bark charcoal of A. mangium was produced in flat kilns
built near the pulp mill. A flat kiln is a built-up brick
furnace with an open top for the continuous addition of
materials (Okimori et al. 2003). This method is often
adopted to char powdered or granular materials such as
bark and sawdust. The charring temperature was low,
ranging from 260 to 360°C. The air-dried charcoal sample
was mixed with distilled water or 1 N 1 mol L−1 KCl at
a ratio of 1:5 to determine the pH (H2O) and pH (KCl),
respectively. The contents of total C and N were deter-
mined by the dry combustion method using an NC-
analyzer 1000, Sumigraph (Shimadzu A-6000, Kyoto,
Japan). The sample, oven-dried at 105°C, was analyzed
to determine the chemical properties as follows. The
dried sample was subjected to extraction with 0.1 N
0.1 mol L−1 HCl and 0.03 N 0.03 mol L−1 ammonium
fluoride to determine the amount of available P2O5 (Bray 1),
with 1 N 1 mol L−1 ammonia acetate to determine the CEC
and amounts of exchangeable cations (K+, Na+, Ca2+
and Mg2+), and with 1 N 1 mol L−1 KCl to determine
the exchangeable acidity (Al3+ and H+). All analyses,
except for the total N and C contents, were carried out
at the Soil Research Institute, Bogor, Indonesia.
Experimental design
The experiments were done twice in two different years,
2003 (Experiment 1) and 2004 (Experiment 2).
Experiment 1: Effect on crop yield
In this experiment, the effect of bark charcoal applica-
tion on the yield of maize (Zea mays L.), cowpea (Vigna
unguiculata [L.] Walp.) and peanut (Arachis hypogaea
L.) was examined at Sites A and B. Three treatments, no
application (Con), application of chemical fertilizer 15-
15-15 (BASF, Aktiengesellschaft, Germany) at 50 g m−2
(NPK), and application of the fertilizer at 50 g m−2 and
bark charcoal at 10 L m−2 (Char-NPK), were conducted
for each crop. The amount of applied chemical
fertilizer, 50 g m−2, which was equivalent to 75 kg ha−1
of each component, was determined in a preliminary
experiment because this amount was found to be effec-
tive in the growth and production of cowpea under
charcoal application. For each treatment, three ridges
measuring 1 m × 2 m in size were prepared and the
applied chemical fertilizer and bark charcoal were
mixed with topsoil at a depth of approximately 10 cm.
For the experiment on maize, nine seed sowing posi-
tions were prepared in the ridge at an inter-row spacing
of 30 cm and an intra-row spacing of 50 cm. For each
position, four seeds were sown, which were thinned to
two plants each after 2 weeks. Thus, 18 plants were
grown in total in each ridge. For the experiment on
cowpea, six seed sowing positions were prepared in
each ridge at an inter-row spacing of 60 cm and an
intra-row spacing of 60 cm. For each position, five
seeds were sown, which were thinned to two plants
each after 2 weeks. Thus, 12 plants were grown in total
in each ridge. For the experiment on peanut, 28 seed
sowing positions were prepared in each ridge at an
inter-row spacing of 20 cm and an intra-row spacing of
20 cm. For each position, three seeds were sown, which
were thinned to one plant each after 2 weeks. Thus, 28
plants were grown in total in each ridge.
The seeds were sown on 19 September 2003 at
Site A and on 1 October 2003 at Site B. For cowpea,
seedpods longer than 40 cm were harvested every 3 days
for 1 month from day 60 after seed sowing. After
3 months, pieces of corn with cob were harvested for
maize, and seedpods were harvested for peanut. The
total fresh weight of the harvested crops was measured
in each ridge for each crop.
Experiment 2: Effects on yield, root amount and
colonization rate of AM fungi in maize
The effects of bark charcoal application on yield, root
amount and colonization rate of AM fungi in maize at
Site C were examined. Four treatments, no application
(Con), application of chemical fertilizer at 50 g m−2
(NPK), application of bark charcoal at 10 L m−2 (Char),
and application of fertilizer at 50 g m −2 and bark
charcoal at 10 L m−2 (Char-NPK), were conducted. For
© 2006 Japanese Society of Soil Science and Plant Nutrition

each treatment, three ridges measuring 1 m × 2 m in size
were prepared and the applied chemical fertilizer and
bark charcoal were mixed with topsoil at a depth of
approximately 10 cm. The number and size of the pre-
pared ridges for each treatment, the method of applica-
tion of the chemical fertilizer and bark charcoal, the
seed sowing position and the number of plants were the
same as those described for maize in Experiment 1.
The seeds were sown on 12 September 2004. The
total fresh weight of harvested corn with cobs was
measured in each ridge at 3 months after seed sowing.
At harvest, roots were collected from four samples of
core topsoil (5 cm in diameter and 10 cm in depth) col-
lected around the center of each ridge. The root samples
in each ridge were gathered to determine the dry weight
and colonization rate of AM fungi. The colonization
rate of the AM fungi was determined for the randomly
collected fine roots using the grid-line intercept method
(Giovannetti and Mosse 1980) after staining with a
0.05% Trypan blue solution.
Soil analyses
Topsoil samples, 10 cm in depth, were randomly collected
from 10 spots at each experimental site, Sites A, B and
C, before the experiments and from three spots in each
Figure 1 Effect of the application of
chemical fertilizer 15-15-15 at 50 g m−2
(NPK) and bark charcoal at 10 L m−2
(Char) on the fresh weight (FW) of
harvested maize, cowpea and peanut
crops. Control (Con): no application.
Values are means ± standard error
(n = 3). Columns with the same letter
(a, b and c) are not significantly
different (P < 0.05).
© 2006 Japanese Society of Soil Science and Plant Nutrition
Effect of charcoal application on crop yield 491
ridge after harvest, which were mixed together at each
experimental site or in each treatment.
For each of the collected soil samples, the chemical
properties, pH (H2O), pH (KCl), amount of available
P2O5 (Bray 1), CEC, amounts of exchangeable cations
(K +, Na +, Ca 2+ and Mg 2+) and exchangeable acidity
(Al3+ and H+) were determined. The analytical methods
were the same as those described above for the charcoal
sample. For the determination of the total N content
(Kjeldahl), the samples, oven-dried at 105°C, were
decomposed in heated sulfuric acid and then distilled
ammonia was collected. All the analyses were carried
out at the Soil Research Institute, Bogor, Indonesia.
Statistical analysis
For the results on crop yield, root dry weight and
colonization rate of AM fungi, one-way anovas were
carried out to evaluate the difference between the
treatments.
RESULTS AND DISCUSSION
The yield of maize, cowpea and peanut at Sites A and B
is shown in Fig. 1. The yield of maize and peanut was
significantly higher after the application of bark charcoal
492 M. Yamato et al.
Figure 2 Effects of the application of chemical fertilizer 15-15-15 at 50 g m−2 (NPK) and bark charcoal at 10 L m−2 (Char) on the
fresh weight (FW) of harvested maize, root dry weight and the colonization rate of arbuscular mycorrhizal (AM) fungi in maize.
Control (Con): no application. Values are means ± standard error (n = 3). Columns with the same letter (a, b and c) are not
significantly different (P < 0.05).
Table 1 Properties of the applied bark charcoal
Available
pH pH Total C Total N P2O 5 CEO
(H2O) (KCl) (kg kg−1) (g kg−1) (mg kg−1) (cmolc kg−1)
7.4 7.1 0.398 10.4 63.1 37.14
and fertilizer (Char-NPK) than after the application of
fertilizer only (NPK) at Site A. Particularly for maize, an
approximate twofold increase in yield was obtained by
bark charcoal application. At Site B, the average yield
of cowpea and peanut in the different treatments
showed the same tendency as that at Site A, namely,
Con < NPK < Char-NPK, although no significant
difference (P < 0.05) was observed. The yield of maize at
Site C is shown in Fig. 2. The yield increased significantly
after Char-NPK application. It was observed that the
standard errors for the means of yield among the repli-
cates were smaller in the Char-NPK application than in
the application of other materials. This suggests that the
yield after Char-NPK application was more stable than
that after a single application of NPK or Char. As the
amount of applied chemical fertilizer, 50 g m−2, which
was equivalent to 75 kg ha−1 of each component, was
approximately half of that of the standard fertilizer
application in the region, it was expected that the
amount of fertilizer applied could be reduced by the
application of bark charcoal.
Exchangeable
Bulk
Exchangeable cations acidity (cmolc kg−1)
density
Ca2+ Mg2+ K+ Na+ Al3+ H+ (g cm−3)
48.82 2.78 4.03 1.85 0.01 0.08 0.37
The applied bark charcoal displayed a neutral pH,
high carbon content, high CEC and high content of
exchangeable cations, particularly Ca2+ (Table 1). The
amount of Ca 2+ was higher than the CEC, which
indicates that free Ca2+ was included in bark charcoal.
Glaser et al. (2000) suggested that the high CEC of
charcoal resulted from slow oxidation on the edges
of the aromatic backbone of charcoal for the formation of
carboxylic groups. Based on the contents of total N and
available P in charcoal (Table 1), charcoal application,
10 L m−2, was equivalent to a total N dose of 38.5 g m−2
and an available P dose of 0.23 g m−2, respectively. A
large amount of total N, approximately fivefold as high
as that applied using chemical fertilizer, was supplied
by charcoal application.
The results of the soil analysis are shown in Table 2.
Before the experiments, the amounts of available P2O5
(Bray 1) and exchangeable cations and base saturation
were higher at Site B than at Sites A or C. The previous
chicken farming activities probably influenced the fer-
tility of the soil at Site B. Soil analysis at harvest revealed
© 2006 Japanese Society of Soil Science and Plant Nutrition
 Effect of charcoal application on crop yield 493

Table 2 Soil chemical properties before the experiment and at harvest

Exchangeable Exchangeable
Available Base
cations (cmolc kg−1) acidity (cmolc kg−1)
Site and pH pH Total N P2O 5 CEC saturation
Treatments (H2O) (KCl) (g kg−1) (mg kg−1) (cmolc kg−1) Ca2+ Mg2+ K+ Na+ (%) Al3+ H+

Site A
Before experiment 3.9 3.9 1.7 47.8 10.12 1.21 0.30 0.15 0.25 18.9 2.67 0.26
At harvest
Maize
Con 4.0 3.8 1.7 22.0 9.87 0.80 0.20 0.12 0.22 13.6 2.93 0.20
NPK 4.0 3.8 1.9 44.5 10.23 0.79 0.18 0.12 0.07 11.3 2.97 0.22
Char-NPK 5.1 4.5 2.4 80.2 12.72 6.21 0.45 0.25 0.13 55.3 0.12 0.12
Cowpea
Con 4.2 3.9 1.2 25.4 10.81 1.30 0.24 0.08 0.08 15.7 2.49 0.15
NPK 4.1 3.9 1.8 44.0 10.66 1.01 0.28 0.12 0.02 13.4 2.81 0.15
Char-NPK 5.3 4.8 1.6 91.8 16.42 7.80 0.43 0.29 0.14 52.7 0.02 0.12
Peanut
Con 4.2 3.9 1.9 55.3 10.21 1.27 0.29 0.08 0.07 16.7 2.41 0.15
NPK 4.0 3.9 1.8 137.5 10.01 0.91 0.22 0.10 0.03 12.6 2.77 0.20
Char-NPK 5.1 4.6 2.5 176.0 12.66 5.98 0.37 0.18 0.07 52.1 0.15 0.10
Site B
Before experiment 4.7 4.1 2.0 210.1 11.77 4.05 1.04 0.40 0.25 48.8 0.48 0.20
At harvest
Maize
Con 4.4 3.9 1.7 23.4 11.64 2.97 0.81 0.22 0.09 35.1 1.64 0.18
NPK 4.4 3.9 2.1 113.2 12.76 3.11 0.73 0.22 0.13 32.8 1.44 0.19
Char-NPK 5.6 5.1 2.4 126.6 15.42 9.32 0.82 0.31 0.13 68.6 0.00 0.02
Cowpea
Con 5.2 4.6 1.9 75.3 11.93 3.77 1.35 0.41 0.13 47.4 0.04 0.11
NPK 4.6 4.1 1.8 151.2 11.23 5.22 1.48 0.22 0.13 62.8 0.44 0.09
Char-NPK 5.8 5.3 2.4 188.2 15.11 9.30 1.10 0.53 0.13 73.2 0.00 0.02
Peanut
Con 4.6 4.1 2.0 302.9 12.31 4.56 1.39 0.59 0.13 54.2 0.26 0.15
NPK 4.5 4.0 2.0 298.7 13.21 4.51 1.19 0.53 0.10 47.9 0.40 0.16
Char-NPK 5.5 5.0 2.8 355.6 17.18 9.73 1.25 0.65 0.20 68.9 0.00 0.05
Site C
Before experiment 4.1 3.9 1.4 94.2 8.54 0.64 0.21 0.07 0.06 11.5 2.35 0.35
At harvest
Maize
Con 4.5 3.9 1.3 108.1 8.85 0.79 0.27 0.07 0.06 13.4 2.31 0.31
NPK 4.4 3.8 1.3 144.5 8.60 0.79 0.25 0.14 0.05 14.3 2.10 0.33
Char 5.4 4.5 2.1 84.3 12.38 5.86 0.55 0.21 0.09 54.2 0.11 0.21
Char-NPK 5.3 4.4 2.1 148.7 13.04 6.06 0.45 0.18 0.06 51.8 0.23 0.17

that the pH, contents of total N and available P2O5,
CEC, contents of exchangeable cations and base satura-
tion were generally higher after the application of bark
charcoal and fertilizer (Char-NPK) than after the application
of fertilizer only (NPK). The high level of available P2O5
after Char-NPK application indicated that the application
of bark charcoal led to a high retention of nutrients. The
increase in base saturation as well as CEC showed that
a large amount of exchangeable cations was introduced
by bark charcoal application. These effects of charcoal
application on soil chemical properties were also reported
by Tryon (1948). In contrast, the amount of exchange-
able Al3+ markedly decreased after the application of
bark charcoal. Low pH, low base saturation, high Al3+
content and low fertility, which are typical characteris-
tics of soils in the tropical region, lead to a low produc-
tivity of crops in this region. Therefore, amelioration of
the soil properties by bark charcoal application could
be effective in increasing crop yield particularly in the
tropical region. The effect of bark charcoal application on
crop yield was not significant at Site B, which indicates
that the effect would not be appreciable on fertile soils.

© 2006 Japanese Society of Soil Science and Plant Nutrition

494 M. Yamato et al.
The effects on the root dry weight and colonization
rate of AM fungi of maize at Site C are shown in Fig. 2.
The root amount significantly increased after the applic-
ation of bark charcoal. Ishii and Kadoya (1994) also
reported an increase in the root amount after the appli-
cation of charcoal. Because it has been shown that char-
coal application increases soil water retention (Piccolo
et al. 1996) and the gaseous phase (Ezawa et al. 2002),
such amelioration of the soil physical and chemical
properties could be effective in enhancing root growth.
The enhancement of root growth may account for
the stable crop production, as evidenced by the lower
standard error in the application of bark charcoal
(Figs 1,2), because enhanced root growth may reduce
the effect of uneven soil conditions. The colonization
rate of AM fungi was highest in the case of bark
charcoal application without fertilizer (Fig. 2). A large
number of studies on the effect of charcoal application
on the enhancement of AM fungal colonization have
been conducted (Ezawa et al. 2002; Ishii and Kadoya
1994; Ogawa 1989; Saito 1990). Ogawa (1989, 1994)
suggested that the porous structure of charcoal may
create a favorable habitat for symbiotic microorganisms.
Further studies on the microenvironment should be
carried out to clarify the relationship between charcoal
application and such symbiosis.
Based on calculations using the bulk density
(0.37 g cm−3) and the carbon content (39.8%) of applied
bark charcoal, the application at 10 L m−2 was found
to be equivalent to a carbon dose of 1.47 kg m−2 (14.7
Mg ha−1). The aromatic structure of charcoal is highly
resistant to abiotic and biotic degradation (Glaser
et al. 2002; Schmidt et al. 1999). Shindo (1991) showed
that charred plant residues applied to volcanic ash
soil were hardly decomposed by microorganisms
even after 40 weeks. In the Brazilian Amazon region,
patchy distribution of black soil containing large
amounts of charred carbon, the so-called Terra Preta,
was found. The soil environment is assumed to be
anthropogenic probably because of charcoal production
in hearths by pre-Columbian Indios (Glaser et al.
2002). The long persistence of charcoal carbon in soil
environments was suggested by the presence of black
carbon for 1000–2000 years in Terra Preta, as indicated
by 14C dating (Glaser et al. 2000, 2002).
In the present study, it was shown that bark charcoal
application is effective in increasing the yield of crops
through the amelioration of the soil chemical properties
and the creation of an appropriate environment for
root growth and AM fungal colonization. Consider-
ing the long persistence of charcoal carbon in the soil
environment, charcoal application in agriculture
could contribute to the formation of a carbon sink in
farmlands.
ACKNOWLEDGMENTS
This study was financially supported by The Kansai
Electric Power Company. We thank Mr Minoru Sugai for
the instructions for charcoal making and Mr Naohiro
Matsui for his help in the analysis of charcoal carbon.
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 PRODUCT RESEARCH - 06/07

Effects of Carbon-Based Soil Amendment (CSA) on Crop Yield of

Irish Potato, Sweet Corn, Tomato and Bell Pepper
By
Ronald Morse, Ph.D. Paul Stevens—Graduate Student
Professor Emeritus, Vegetable Crops Department of Horticulture, VA Tech
Department of Horticulture, VA Tech Blacksburg, VA 24061-0327
Blacksburg, VA 24061-0327

Summary

This research evaluated preplant in-row application of CSA on crop yield of Irish potato, sweet
corn, tomato and bell pepper at the Kentland Agricultural Research Farm, located near
Blacksburg, VA. Overall, our data are encouraging, indicating that beneficial microorganisms
(microbes) in CSA granules improved productivity of sweet corn (2006 and 2007) and tomato
(2007). Based on limited data and observations, enhanced mineralization in the rhizosphere of
CSA-treated sweet corn and tomato plants is at least partially responsible for increased uptake of
nitrogen and improved crop yield. Apparently, placement in near proximity to sweet corn and
tomato seed stimulated rapid rhizosphere development of microbial biomass and improved
effectiveness of CSA. Crop yield of Irish potato and bell pepper was not affected by CSA. Distant
placement of CSA (3-4 inches above the seed pieces in 2007) possibly accounted for lack of crop
yield response with Irish potato. Reasons for lack of yield response of CSA to bell pepper are not
know, since tomato and pepper transplants were treated the same throughout these experiments.

Materials and Methods

Description of field soil and treatments. In 2006 and 2007, all field plots were grown in an
organic transition high-quality silt loam soil having a pH of 6.4, with medium to high levels of
phosphorus, potassium, calcium and magnesium.

Irish potato and sweet corn. The research field plots both years were divided into four
replications (424 ft long and 24 ft wide), consisting of four raised beds (6 ft center to center, and
6 inches high). The top of each bed was approximately 42 inches wide, and the alleyways
between beds (bed shoulders and bottoms) were 30 inches. In September preceding each year,
two beds in each replication were seeded with forage radish (Raphanus sativus) and two beds to
crimson clover (Trifolium incarnatum). Alleyways in all plots were seeded with cereal rye
(Secale cereale). Forage radish was frost killed at approximately 20 F and crimson clover and
rye over-wintered. In mid April, bed tops were flail mowed and the residues were shallow
incorporated (2-3 inches deep) with a rototiller (Table 1).

Biomass produced (and thus mineralizable nitrogen, N) of both cover crops was considerably
less in 2006/2007 than 2005/2006. Although actual amounts of cover crop biomass and
mineralizable N were not measured, the estimated quantity of plant-available N would be
approximately 30 and 15 lb N/acre in 2006, compared to 10 and 5 lb N/acre in 2007, from
crimson clover and forage radish, respectively. Because nitrogen availability was a major
limiting factor both years, especially in 2007, these relatively small amounts of plant-available N
would have contributed relatively little to improved crop yield. In mid April of 2006 (but not in
2007), before seeding Irish potato or sweet corn, all beds were fertilized using 800 lb/acre of
Renaissance 8N-1P-5K organic fertilizer (composed of feather meal, bone meal, soybean and
potassium sulfate). The organic fertilizer was precision placed in-row and incorporated to a depth
of 5-6 inches deep in grow zones located 20-inches apart on top of the raised beds. Grow zones
are the designated row areas on raised beds where vegetables were seeded.

Growing Irish potato. Whole seed potatoes (Chieftain in 2006 and Kueka Gold in 2007) were cut
into 2-oz seed pieces, treated with Oxidate for 5 minutes, stored for 1-2 weeks and planted by
hand in 2006 and using a potato seeder in 2007 (Table 1). Seed pieces were set 10-12 inches
apart and 5-6 inches deep in the two grow zones on each bed (212 ft long). In 2006, CSA
granules (30 lb/acre) were applied before planting the seed pieces. CSA granules were hand
applied to subplots (two beds, 76 ft long) in the grow zones and incorporated to a depth of 3-5
inches with a wheel hoe. In 2007, CSA granules (20 lb/acre) were applied (after planting potato
seed pieces with the seed planter) in the grow zones and shallow incorporated (only 1-2 inches
deep) with a hand hoe. Each year, the remaining row area (136 ft long) of each row was left
untreated, of which the middle 60 ft was designated as a buffer zone separating the CSA-treated
(+CSA, 76 ft) from the untreated (no CSA, 76 ft). Plots were irrigated and hand weeded as
needed throughout the growing season. Stand counts were made and potato tubers were
harvested in late September using a two-row potato digger. In 2007, each subplot was further
divided into two equal sub-subplots (one 6-ft bed, 76 ft long, received 2 gallon of fish
concentrate/acre and a second adjacent bed received no fish concentrate—i.e., control). Each
sub-subplot was further divided into two equal sub-sub-subplots (one bed, 38 ft long) (one-half
of each sub-subplot received 80 lb N/acre as a sidedress fertilizer and the other half received no
sidedress fertilizer—i.e., control).

Growing sweet corn. Sweet corn seed (Spring Treat in 2006 and Sugar Queen in 2007) was hand
planted with and Earthway Seeder in the two grow zones (212 ft long and 20 inches apart) of the
raised beds. Immediately before seeding in both years, CSA granules were applied to subplot in-
row grow zones (76 ft long) of each bed. In 2006, CSA granules (20 lb/acre) were hand applied
and incorporated to 1-2 deep with a wheel hoe. In 2007, CSA granules (30 lb/acre) were hand
applied using the Earthway Seeder to a depth of 1-2 inches. The remaining area (136 ft) of each
bed was left untreated, of which the middle 60 ft was designated as a buffer zone, separating the
CSA-treated (+CSA, 76 ft) and the untreated (no CSA, 76 ft) sections. Plots were irrigated and
hand weeded as needed throughout the growing season. Stand counts were made and sweet corn
ears were harvested by hand (Table 1). In 2007, subplots were further divided into sub-subplots
and sub-sub-subplots, as described above for Irish potato.

Nitrogen sidedressing (2006). Two weeks after immergence, potato plants in untreated subplots
(no CSA) were sidedressed by hand at the rate of 60 lb N/acre (20 lb N from sodium nitrate and
40 lb N from feather meal). Plants in the CSA-treated (+CSA) were not sidedressed. Five weeks
after planting, sweet corn plants of both +CSA and no-CSA subplots were divided into three sub-
subplots (25 ft long) and were sidedressed by hand at three rates of nitrogen fertilizer (0, 50 and
100 lb N/acre, from a mixture of sodium nitrate and feather meal).
Tomato and bell pepper (2007). In two separate experiments, tomato (Mountain Fresh) and bell
pepper (Aristotle) were grown on small plastic covered plots (24 ft wide and 72 ft long). The
experimental design was a randomized split block, with four replications. Main plots were
transplant-growing mixes: McEnroe Lite (ML—an organic potting mix) and Metro Mix 360
(MM—an inorganic potting mix). Subplots were addition of CSA granules to the growing mixes
(1 cup CSA granules/5 gallons of growing mix): untreated control (no CSA) and CSA-treated
(+CSA—granules were thoroughly blended in the growing mixes). Seeds of tomato or bell
pepper were placed into 72-cell trays containing either untreated or CSA-treated ML or MM
growing mixes, and grown to maturity (about 7 weeks for tomato and 9 weeks for bell pepper).
At maturity, the tomato and bell pepper transplants were set by hand in previously established
raised beds covered with black plastic mulch. A starter solution consisting of hydrolyzed fish
concentrate (2 gallons/acre) was applied with water (200 gallon/acre) as a liquid drench around
the base of each transplant in all treatments. To avoid contamination, untreated (no CSA) and
CSA-treated subplots were separated by 150 ft.

A cereal rye/hairy vetch (R/HV) cover crop biculture was seeded in the tomato/bell pepper field
site in early October of 2006. Before laying off raised beds and applying black plastic mulch, the
R/HV cover crop was flail mowed and incorporated using a disk plow. No fertilizer was applied
at planting or during the growing season for either tomato or bell pepper; however, the
incorporated R/HV residues would have provided approximately 40-60 lb N/acre.

Results and Discussion

Effects of CSA on marketable crop yield. Marketable crop yield was highest in CSA-treated
plots for sweet corn (2006 and 2007) and tomato (2007). Application of CSA had no crop yield
effects for Irish potato and bell pepper (Table 2). Why different yield responses to CSA occurred
for tomato (17% increased fruit yield) and bell pepper (no response) is unknown, since both
crops were treated identically throughout the duration of the experiments. In-row placement of
CSA granules was distinctly different for Irish potato and sweet corn and could possibly account
for different yield responses to application of CSA. Granules of CSA were placed in close
proximity to sweet corn seed in both 2006 and 2007; however, in 2007, shallow incorporation of
CSA granules and deep placement of potato seed pieces (5-6 inches below the soil surface)
resulted in relatively reduced CSA-seed contact with potato, compared to sweet corn. In 2006,
CSA-seed contact was adequate; however, yield response to application of CSA granules was
confounded, because the untreated control (no CSA) potato plants were sidedressed with 60 lb
N/acre, while the CSA-treated plants were not sidedressed.

How (what mechanisms) CSA improves crop yield is mere speculation. Perhaps, CSA microbes
(at proper concentration, placement and timing) can improve absorption of plant-available soil
moisture, and/or improve rate of mineralization (release of plant-available N). Drip irrigation
was applied uniformly across all plots both years; hence, we have no evidence that improved
water absorption contributed to improved crop yield in CSA-treated plots. On the other hand,
enhanced mineralization could have accounted for the increase crop yields, as evidence by the
potato data for 2006. Potato tuber yield in 2006 was nearly identical in CSA-treated and untreated
(no CSA) plots, although the untreated plants received 60 lb N/acre as a fertilizer sidedressing,
while CSA-treated plants were not sidedressed. Since the soils in our experimental sites were
nitrogen deficient, these data indicate that more plant-available N was released (mineralized) in
CSA-treated than untreated plots.
Effect of growing mixes and CSA on marketable fruit yield of tomato. An interaction
occurred between potting mixes used for growing tomato transplants and application of CSA
granules. Tomato transplants grown in the inorganic Metro Mix 360 (MM) out yielded plants
grown in the organic McEnroe Lite (ML) by 6%; however, when CSA granules were added to
the potting mixes, tomato transplants grown in MM out yielded ML by 16% (Table 3).
Apparently, proliferation of beneficial microbes was uninhibited in the more “sterile” CSA-
treated MM, while microbial competition possibly reduced buildup of beneficial microbes in
CSA-treated ML. The result was a 22% tomato yield increase in CSA-treated MM (vs. untreated),
compared to only a 12% yield increase in CSA-treated ML (Table 3).

Effect of CSA on growth of tomato and bell pepper transplants. Application of CSA granules
to MM and ML potting mixes (priming) enhanced growth rate and size of both tomato and bell
pepper. Although not measured, transplant size at time of field setting was approximately 15-
25% larger. Enhancement in growth response to CSA priming appeared to be greater for tomato
than bell pepper. The potential of CSA priming to shorten the time required to produce
marketable transplants and subsequent increased marketable fruit yield merits further research.
(Refer to the 2006 report).

Effect of cover crops on marketable yield of Irish potato and sweet corn. There was no yield
response to cover crops in 2007 (data not shown). No yield response is highly predictable, since
growth of cover crops was severely curtailed because of delayed seeding, drought and poor plant
stands. Refer to the 2006 report for discussion of the cover crop effects on crop yield in 2006,
when growth of cover crops was excellent.

Effect of N sidedressing and liquid fish concentrate on marketable yield of Irish potato and
sweet corn in 2007. In accordance with the agreed-upon protocol, no preplant fertilizer was
applied in 2007. Therefore, since the soil at the research sites is relatively low in plant-available
N and little cover crop biomass was produced, these unfertilized plots showed a dramatic yield
response to N sidedressing (80 lb N/acre) for both Irish potato and sweet corn (Table 4), and
even showed a slight response to in-row application of liquid fish concentrate for Irish potato
(Table 5).

Table 1. Dates of important cultural practices.

Cultural practice 2006 2007

Seeded cover crops Sept. 6 (05) Sept. 18 (06)

Applied preplant fertilizer Mid April ------

Planted Irish potato April 19 May 9

Planted Sweet corn June 13 May 31

Transplanted tomato and bell pepper ------ June 11

Applied nitrogen sidedressing—Irish potato* May 24 June 13

Applied nitrogen sidedressing—sweet corn July 18 July 2

*Only untreated (no CSA) potato plants were sidedressed (60 lb N/acre) in 2006.
Table 2. Effect of application of CSA on marketable organic crop yield.
2006 2007
Vegetable Yield (cwt/acre) Yield (cwt/acre)
Crop No CSA CSA Sign. No CSA CSA Sign.

Irish potato 189* 183 ns 88 82 ns

Sweet corn 93 107 .05 61 67 .10

Tomato ---- ---- ---- 634 740 .10

Bell pepper ---- ---- ---- 352 357 ns

*In 2006, untreated (no CSA) potato plots received 60 lb N/acre as a sidedressing; CSA-treated
potato plots were not sidedressed.
Cwt = hundred weight units (100 lb); ns = not statistically significant at p = .10

Table 3. Effect of priming growing mixes with CSA on marketable organic crop yield, 2007.

Growing mix Yield (cwt/acre) Difference (cwt/acre)

No CSA CSA Avg. CSA- no CSA (%)
Tomato

ML = McEnroe Lite 615 687 651 +72 +12

MM = Metro Mix 360 653 794 724 +141 +22

Avg. 634 740 ---- +106 +17

Difference: MM-ML +38 +107 +73 ---- ----

(%) +6 +16 +11 ---- ----

Bell Pepper

ML = McEnroe Lite 366 368 367 +2 0

MM =Metro Mix 360 339 346 342 +7 +2

Avg. 352 357 ---- +5 +1

Difference: MM-ML -27 -22 -25 ---- ----

(%) -7 -6 -6 ---- ----

CSA = Carbon Based Soil Amendment; cwt = hundred weight units (100 lb); Avg. = average;
CSA-no CSA = CSA minus no CSA; MM-ML = MM minus ML.
Table 4. Effect of nitrogen fertilizer sidedressing on marketable organic crop yield of Irish
potato and sweet corn.

2006 2007

Vegetable N rate Yield N rate Yield

Crop (lb/acre) (cwt/acre) (lb/acre) (cwt/acre)

Irish potato ----- ----- 0 73

----- ----- 80 98
Sign. .001

Sweet corn 0 102 0 31

50 98 80 96

100 100 Sign. .001

Sign. ns
Cwt = hundred weight units (100 lb); ns = not statistically significant at p = .05.

Table 5. Effect of liquid fish concentrate applied in row at planting on marketable organic
crop yield of Irish potato and sweet corn, 2007.

Liquid fish Yield (cwt/acre)

(Gallons/acre) Irish potato Sweet corn

0 80 61

2 91 66

Significance .05 ns
Cwt = hundredweight units (100 lb); ns = not statistically significant at p = .05.
Biochar, climate change and soil:
A review to guide future research

Saran Sohi1, Elisa Lopez-Capel2, Evelyn Krull3 and Roland Bol4

Corresponding author and editor: Evelyn Krull
CSIRO Land and Water Science Report 05/09
February 2009
CSIRO Land and Water Science Report series ISSN: 1834-6618

Copyright and Disclaimer

© 2008 CSIRO To the extent permitted by law, all rights are reserved and no part of this
publication covered by copyright may be reproduced or copied in any form or by any means
except with the written permission of CSIRO.

Important Disclaimer:
CSIRO advises that the information contained in this publication comprises general
statements based on scientific research. The reader is advised and needs to be aware that
such information may be incomplete or unable to be used in any specific situation. No
reliance or actions must therefore be made on that information without seeking prior expert
professional, scientific and technical advice. To the extent permitted by law, CSIRO
(including its employees and consultants) excludes all liability to any person for any
consequences, including but not limited to all losses, damages, costs, expenses and any
other compensation, arising directly or indirectly from using this publication (in part or in
whole) and any information or material contained in it.

Citation: Sohi, S1.Loez-Capel, E2., Krull, E3., Bol, R4., 2009. Biochar's roles in soil and climate
change: A review of research needs. CSIRO Land and Water Science Report 05/09, 64 pp..
1
Centre for Bioenergy and Climate Change, Rothamsted Research, Harpenden, AL5 2JQ, UK; now at
UK Biochar Research Centre, School of Geosciences, University of Edinburgh, UK, Email:
saran.sohi@ed.ac.uk
2
School of Civil Engineering and Geosciences, University of Newcastle, Newcastle uponTyne, NE1 7RU,
UK, Email: elisa.lopez-capel@ncl.ac.uk
3
CSIRO Land and Water, PMB2, Glen Osmond SA 5064, Australia, Email: evelyn.krull@csiro.au
4
Biogeochemistry of Soils and Water group, North Wyke Research, UK, Okehampton, EX20 2SB, UK,
Email: roland.bol@bbsrc.ac.uk

Cover Photographs:
Cover photo 1 Fragments of biochar 0.2-0.5mm diameter comprising willow stems carbonised at 600
C. Source: Saran Sohi

Cover photo 2 Scanning electron micrograph of biochar formed from rubber tree stems at
800 C in a 200kW gasification plant in rural Cambodia. Source: Simon Shackley and Erik
Middelink

© 2008 CSIRO
ACKNOWLEDGEMENTS

The preparation of this report was funded by the CSIRO Land and Water Opportunity
Development Fund.
Rothamsted Research and North Wyke Research received grant-aided support from the
Biotechnology and Biological Sciences Research Council (BBSRC) in the UK. Rothamsted
Research is an Institute of the UK Biotechnology and Biological Sciences Research Council.
Newcastle University receives grant-aided support from the Engineering and Physical
Sciences Research Council (EPSRC) in the UK.
Keith Goulding (Manager of the Cross-Institute Programme for Sustainable Soil Function
Department of Soil Science, Rothamsted Research), Peter Hairsine (Leader, Science
Development CSIRO Land and Water) and Mike Grundy (Theme Leader, Managing
Australia’s Soil & Landscape Assets. CSIRO Land and Water) provided helpful reviews and
comments that improved the final version of the report.

iii
EXECUTIVE SUMMARY
Biochar is the charred by-product of biomass pyrolysis, the heating of plant-derived material
in the absence of oxygen in order to capture combustible gases. The objective of this report
was to review and evaluate published studies with regard to what evidence and arguments
currently exist that assess the application of biochar to soil to a) sequester carbon and b)
produce secondary agronomic benefits. Current analyses suggest that there is global
potential for annual sequestration of atmospheric CO2 at the billion-tonne scale (109 t yr-1)
within 30 years. So far, however, the underlying published evidence arises mainly from
small-scale studies that do not currently support generalisation to all locations and all types
of biochar.

From the available published and peer-reviewed literature the following general findings can
be made for eight key questions. It is noted that for each of these key questions there
remains major research questions that require the attention of researchers. This summary
provides these outstanding research issues along side the major findings.

1. Is all biochar the same?

Key chemical and physical properties of biochar are greatly affected both by choice of
feedstock (crop waste, energy crop, wood chip, municipal waste, manure, etc.) and
process conditions (mainly temperature and time). These properties affect the
interactions biochar has within the environment of its application as well as its fate. A
rapid screening technique that provides the means for biochar products to be compared
or matched to a particular use does not currently exist.

2. How stable is it?

Studies of charcoal from natural fire and ancient anthropogenic activity indicate
millennial-scale stability. However, it is difficult to establish the half-life of modern biochar
products using short experiments due to the presence of small amounts of labile
components, partial oxidation and biotic or abiotic surface reactions. At the moment
there is no established method to artificially-age biochar and assess likely long-term
trajectories.
3. Is it safe to use?
Analysis of a limited number of biochar samples has indicated concentrations of toxic
combustion products such as polycyclic aromatic hydrocarbons that are not at
environmental risk level. However, a more systematic evaluation for a more complete
range of other potentially harmful chemical contaminants associated with combustion, as
well as toxic substances within feedstocks, has not been made. An environmental risk
assessment that includes the impacts of these on terrestrial or aquatic ecosystems is
outstanding. Given the stability of biochar, safe rates of applications need to be
determined for individual soil types to avoid possible detrimental effects due to over-
application (e.g. reduction in soil productivity).
4. What are the agronomic benefits?
Currently, a large number of studies have been conducted where biochar application has
shown significant agronomic benefits with a minor number of studies showing no
significant effects of biochar application on crop productivity and some studies reporting
adverse effects. This suggests that the extent of the effect of biochar on crop productivity
is variable, due to the different bio-physical interactions and processes that occur when
biochar is applied to soil, which are not yet fully understood.. There is a need for models
to allow extrapolation of location-specific findings by accounting for mechanistic effects
of geographic variations in soil type, climate, cropping and pyrolysis feedstock.
5. Is it economically viable?
The economic value of sequestered carbon will be determined within complex carbon
markets that are influenced by energy supplies and demand, the supply and demand for

iv
 low emissions technologies, the availability of alternative carbon sequestration
technologies and global policy responses to climate change. The growing price of waste
disposal is likely to make the production and application of biochar for electricity and
waste management economically viable. Carbon offsets will have a greater role once
biochar is certified under the Clean Development Mechanism (CDM) of the Kyoto
Protocol. Uncertainty over market interventions may risk the investment in energy
facilities that are able to produce biochar.
6. What are the environmental and societal benefits?
Models exist for viable agronomic use of biochar (without energy capture) in subsistence
agriculture. However, relevant technological innovation is required to deal with
environmental issues such as smoke production from traditional char production and
associated health risks. Socio-economic constraints are not adequately researched.
Higher crop yields resulting from biochar applications would be expected to mitigate
pressures on land and would also have relevance to land restoration and remediation.
Other environmental benefits of biochar application may lie in its application to
rehabilitate contaminated wetlands and as a means to assist in managing (avoiding)
algal blooms in aquatic ecosystems through adsorption of nutrients.

7. Are the benefits of biochar in mitigating greenhouse gases widely accepted?

It is generally accepted that biochar is a highly stable form of carbon and as such has the
potential to form an effective C sink, therefore sequestering atmospheric CO2. Several
projects are currently underway assessing/monitoring greenhouse gas emissions in fields
amended with biochar in USA, Colombia, Kenya and Australia
(<http://www.css.cornell.edu/faculty/lehmann/research/biochar/biocharproject.html>).
While biochar surpasses other biological forms of C with regard to its stability, estimates
on the mean turnover time of biochar in soil vary from hundreds to thousands to tens of
thousands of years. The mitigation potential of biochar with regard to other greenhouse
gases, such as N2O and CH4, through its application to soil is less well established and
requires further research.

8. What are the research gaps and future challenges?

Biochar performance
Currently, the predictive capacity for biochar ‘performance’ does not exist and how to
best optimise the multiple useful characteristics as a function of feedstock has not been
assessed. This is currently inhibiting the realisation and application of this technology.

Interaction of biochar with soil microbial communities and plants

The physical, biological and chemical processes that biochar may exert on microbial
communities and their symbiotic interaction with plants, and possibly enhanced nutrient
use efficiency, are not yet understood. The apparent contradiction between the high
stability of biochar, soil organic matter accumulation and apparent enhancement of soil
microbial activity needs to be resolved. Research in Japan and in Germany has
indicated that biochar can complex the carbon from dead micro-organisms. Further
research work is required to determine under what conditions this complexation takes
place.

Cation exchange capacity (CEC)

While the CEC of fresh char itself is not very high, biochar that has resided in soil for
hundreds of years has been shown to have much higher CECs, comparable to those of
zeolites. However, several studies have reported an increase in soil CEC after the
application of fresh biochar. Thus, the processes that are instrumental in developing
CEC over time as well as the effects that lead to an increase in CEC by addition of fresh
(low CEC) biochar require detailed understanding.

v
Water holding capacity and aggregate stability
While some studies report positive effects of biochar application on water-holding
capacity, the specific mechanism that biochar exerts on water retention, macro-
aggregation and soil stability are poorly understood – yet should be of critical importance
in climate change adaptation, where mitigating drought, nutrient loss and erosion are
critical.

Erosion, transport and fate

The loss of biochar through vertical or lateral flow is not quantified, and only recently
have studies been initiated to examine movement through soil profiles and into
waterways. These processes complicate the task of confining the range of current
estimates (from hundreds of years to millennia) of the mean residence time of biochar in
soil. Long-term monitoring research stations (analogous to the Waite permanent rotation
trial in Australia or the Rothamsted long-term experiments in the UK) are required to
adequately assess the long-term stability and dynamics of biochar in soil.

Decreased emissions of non-CO2 greenhouse gases (e.g. N2O and CH4)

The currently available data on the effect of biochar additions on trace gas emission is
very limited, but has a potentially great impact on the net benefit of biochar application.
Development of cost effective means of measuring decreased emissions will ensure this
potentially large greenhouse saving can be compliant with emissions trading schemes.

Soil carbon modelling

Modelling of the linked carbon and nitrogen cycles in soil with and without application of
biochar is essential to understanding the fundamental mechanisms referred to above
and the impact on soil-based emissions of greenhouse gases.

Project specific Life Cycle Assessment (LCA)

The total environmental life-cycle assessment has been conducted for some biochar
case studies. Greenhouse balances, for example, are very project specific and hence
there is opportunity to assess the benefits over a large range of feedstock, process and
biochar application scenarios.

vi
CONTENTS

1. Introduction ......................................................................................................... 1
2. Biochar ................................................................................................................. 2
2.1. What is biochar? ....................................................................................................... 2
2.1.1. Why and how is biochar made?.............................................................................3
2.1.2. Biochar feedstocks ................................................................................................5
2.1.3. Biochar production systems ..................................................................................6
2.1.3.1. Slow pyrolysis........................................................................................................7
2.1.3.2. Fast pyrolysis.........................................................................................................8
2.1.3.3. Intermediate pyrolysis............................................................................................9
2.1.3.4. Carbonisation ........................................................................................................9
2.1.3.5. Gasification............................................................................................................9
2.1.3.6. Production of ammonia during pyrolysis ..............................................................10
2.2. Biogeochemical characterisation of biochar ........................................................... 10
2.2.1. The purpose of biogeochemical characterisation ................................................10
2.2.2. Physical and chemical characterisation...............................................................11
2.2.3. Quantification of biochar and char .......................................................................13
2.2.4. Background and biochar history: from terra preta to terra nova...........................14

3. Biochar application in Agriculture................................................................... 16

3.1. Biochar and climate change ................................................................................... 16
3.1.1. Soil organic matter and climate change...............................................................17
3.1.2. Carbon stabilisation and sequestration using biochar .........................................19
3.1.3. Combined bioenergy and biochar production ......................................................20
3.1.4. Evaluation of biochar systems .............................................................................21
3.1.5. Stability of biochar in soil .....................................................................................21
3.1.5.1. Stability of biochar in natural systems .................................................................22
3.1.5.2. Influence of biochar feedstock on stability ...........................................................23
3.1.5.3. Climatic effects on biochar mineralisation ...........................................................23
3.1.5.4. Soil biological activity and the stability of biochar ................................................24
3.1.5.5. Effects of biochar on ease of tillage and mechanical disturbance .......................24
3.1.6. Indirect impacts of biochar on CO2-equivalent emissions....................................25
3.1.6.1. Indirect effects of yield benefits and nutrient requirement ...................................25
3.1.6.2. Impact of biochar on nitrous oxide emission from soil .........................................26
3.1.6.3. Impact of biochar on methane emission from soil................................................27
3.1.6.4. Biological activity and stabilisation of soil organic matter ....................................27
3.1.6.5. Effects of biochar on tillage and irrigation requirements ......................................28
3.1.7. Biochar scenarios for agriculture .........................................................................28
3.1.8. Notes on the natural cycling of char in soil ..........................................................31
3.2. Biochar, crop productivity and resource management ........................................... 32
3.2.1. Soil fertility ...........................................................................................................32
3.2.2. Crop yield ............................................................................................................33
3.2.3. Soil moisture retention.........................................................................................35
3.2.4. Nutrient retention and use-efficiency ...................................................................35
3.2.5. Use of biochar to manage water quality ..............................................................37
3.2.6. Potential risks to soil and water from use of biochar............................................37

4. Policy context and Analysis ............................................................................. 38

4.1. A framework to evaluate applications of biochar .................................................... 38
4.2. Scenarios for the uptake of biochar for use in soil.................................................. 38
4.3. Market intervention and carbon trading .................................................................. 39
4.4. Market acceptability issues..................................................................................... 40
4.5. Research................................................................................................................. 41

vii
5. Research Priorities and Future Challenges .................................................... 41
5.1. Fundamental mechanisms...................................................................................... 42
5.2. Properties, qualities and environmental risk assessment....................................... 43
5.3. Carbon cycle modelling .......................................................................................... 44
5.4. Beneficiaries ........................................................................................................... 44
5.5. Commentary on likely barriers to the adoption of a large scale enterprises utilising
Biochar.................................................................................................................... 45
References .................................................................................................................. 46

TABLE OF FIGURES

Figure 1. Biochar as a C sequestration tool

Figure 2. Summary of pyrolysis processes
Figure 3. Illustrative slow pyrolysis process
Figure 4. Schematic process diagram for fast pyrolysis
Figure 5. Simplified flow chart of how biomass releases energy as it captures CO2 as
ammonium carbonate
Figure 6. Scanning electron microscope image of biochar from pelletised peanut shell
Figure 7. Components of black carbon assessed by available quantification methods
Figure 8. Comparison of profiles of terra preta and adjacent soils
Figure 9. The key physical (purple arrows), natural (orange arrows) and anthropogenic (red
arrows) interactions of biochar in the environment
Figure 10. Scenario for multi-feedstock production of biochar, and multi-application use,
emphasising the spatial context

TABLE OF TABLES

Table 1. Fate of initial feedstock mass between products of pyrolysis processes

Table 2. Reported elemental composition for a range of bio-oil and biochar products
Table 3. Properties of biochar from bagasse carbonisation
Table 4. Physico-chemical properties of terra preta and adjacent soils
Table 5. Summary of experiments assessing the impact of biochar addition on crop yield

viii
1. INTRODUCTION
Periodic fire across Australian landscapes results in a natural process of carbon (C)
sequestration from atmosphere to soil by the conversion of biomass to charcoal. The extent
of this process has recently been quantified for Australia (Lehmann, 2009). There are
increasing calls to mirror and enhance this process by the concerted use of ‘biochar’, a form
of charcoal produced with the simultaneous production and capture of bio-energy which is
then applied to the soil. A measure of the need and interest for a concerted effort in this area
has been the evolution of an organised consortium, known as the International Biochar
Initiative (IBI) (<www.biochar-international.org>).

The inspiration for the supplementation of soil with charcoal stems from observations made
in the ancient agricultural management practices that created terra preta, deep black soils.
These soils, found throughout the Brazilian Amazon, are characterised by high levels of soil
fertility compared with soils where no organic C addition occurred (Harder, 2006; Marris,
2006; Lehmann, 2007a; Renner, 2007). The evident value of the terra preta led to the
suggestion that investment into biochar and application to agricultural soil may be both
economically viable and beneficial. Rising fossil fuel prices, the need to raise yields in light of
the global food crisis, and the emergence of a significant global market for trading carbon
appear to promise added economic incentives in the future.
At the same time the need to protect soils under an increasingly uncertain climate makes the
apparent ability of biochar to increase the capacity for soil to absorb and store water vitally
important. It also appears that adding biochar to soil may be one of the only ways by which
the fundamental capacity of soils to store and sequester organic matter could be increased.
There are a number of detailed reviews describing charcoal formation (Knicker, 2007) and
associated C dynamics (Preston et al., 2006; Czimczik et al., 2007), including its role in the
global carbon cycle (Schmidt et al., 2000). Forthcoming is a compendium of review articles
(“Biochar for Environmental Management: Science and Technology”), which will place
existing studies in the context of pyrolysis bioenergy (Lehmann and Joseph, 2009b ).
A number of studies have now highlighted the net benefit of using biochar in terms of
mitigating global warming and as an active strategy to manage soil health and productivity
(Figure 1) (Lehmann, 2007a; Lehmann, 2007b; Lehman et al., 2005; Ogawa et al., 2006;
Laird, 2008; Mathews, 2008; Woolf, 2008). However, relatively few studies exist that make a
quantitative assessment of biochar-based soil management scenarios with regard to
greenhouse gas, energy, and economic perspectives (Fowles, 2007; Gaunt et al., 2008).
Nonetheless, the concept and value of biochar production and application is gradually
incorporated by policy makers and governments (Winsley, 2007).
Current studies are in many cases conceptually or geographically limited, and are often
constrained by limited experimental data. In particular, mechanistic descriptions of the
characteristics of biochar and its function in the soil and experimentation relevant to wide-
scale applications of biochar are currently limited. In this report we examine existing
published research within a framework constrained by a policy context. Thereby, we aim to
identify gaps where new research should be focused in a way that will enable biochar to
engage with climate change mitigation and to maintain soil productivity.

1
 Fossil fuel Bioenergy Bioenergy with
biochar to soil

atmospheric carbon dioxide

combustion combustion pyrolysis

Figure 1. Biochar can

soil carbon result in a net removal
biochar
of carbon from the
atmosphere, especially
with enhanced net
fossil carbon primary productivity

2. BIOCHAR

2.1. What is biochar?

Biochar is a fine-grained and porous substance, similar in its appearance to charcoal
produced by natural burning. Biochar is produced by the combustion of biomass under
oxygen-limited conditions. The definition adopted by the International Biochar Initiative (IBI)
furthermore specifies the need for purposeful application of the material to soil for agricultural
and environmental gain.
The term biochar was originally associated with a specific type of production, known as ‘slow
pyrolysis’. In this type of pyrolysis, oxygen is absent, heating rates are relatively slow, and
peak temperatures relatively low (Section 2.1.3.1). However, the term biochar has since
been extended to products of short duration pyrolysis at higher temperatures known as ‘fast
pyrolysis’ (Section 2.1.3.2) and novel techniques such as microwave conversion.
It is important to note that there is a wide variety of char products produced industrially. For
applications such as activated carbon, char may be produced at high temperature, under
long heating times and with controlled supply of oxygen. In contrast, basic techniques for
manufacture of charcoal (such as clay kilns) tend to function at a lower temperature, and
reaction does not proceed under tightly controlled conditions. Traditional charcoal production
should be more accurately described as 'carbonisation' (Section 2.1.3.4), which involves
smothering of biomass with soil prior to ignition or combustion of biomass whilst wet. Drying
and roasting biomass at even lower temperatures is known as ‘torrefaction’ (Arias et al.,
2008).
A charred material is also formed during 'gasification' of biomass, which involves thermal
conversion at very high temperature (800°C) and in the partial presence of oxygen (Section
2.1.3.5). This process is designed to maximise the production of synthesis gas (‘syngas’).
Materials produced by torrefaction and gasification differ from biochar in physico-chemical
properties, such as particle pore size and heating value (Prins et al., 2006) and have
industrial applications, such as production of chemicals (methanol, ammonia, urea) rather
than agricultural applications.
In order to differentiate biochar from charcoal formed in natural fire, activated carbon, and
other black carbon materials, the following list of terms aims to better define the different
products. The differences, however, are relatively subtle since all products are obtained from
the heating of carbon-rich material.

2
• Char: the solid product arising from thermal decomposition of any natural or synthetic
organic material. Examples are char from forest fire and soot resulting from the
incomplete combustion of fossil hydrocarbon.
• Charcoal: produced from the thermal decomposition of wood and related organic
materials, mainly for use as an urban fuel for heating and cooking, but also traditional
uses as soil amendment or control of odour (Okimori et al., 2003). Temperatures in
traditional kilns approach 450-500°C, which is similar to that of industrial pyrolysis but
with lower yields: conversion of feedstock dry mass may be as low as 10 % compared to
35% using more formal production technology. Also, all heat as well as gaseous and
liquid co-products are lost during the combustion process.
• Activated carbon: manufactured by heating carbonaceous material at a high temperature
(above 500°C) and over long (>10 hours) periods of time. The resulting material is
characterised by a very high adsorptive capacity. It is not used as a soil amendment but
has been applied for cleansing processes, such as water filtration and adsorption of gas,
liquid or solid contaminants (Tomaszewski et al., 2007).
• Black carbon: a general term that encompasses diverse and ubiquitous forms of
refractory organic matter that originate from incomplete combustion (Baldock et al.,
2002). The diversity of burning conditions results in black carbon occupying a continuum
of material. The review by Schmidt (1999) provides a thorough account of the ‘black
carbon’ continuum, its constituents and definitions.
Biochar from pyrolysis and conventional charcoal and char share key characteristics which
are related to carbon sequestration (long residence time) and soil fertility (soil conditioning
effect). This is important since there is currently a much greater amount of research for char
(Glaser et al., 2002) than for biochar. Biochar produced in association with bioenergy
generation may be more applicable in some countries than others, depending on economic
circumstance, political priorities, technology and infrastructure.
The central quality of biochar and char that makes it attractive as a soil amendment is its
highly porous structure, potentially responsible for improved water retention and increased
soil surface area. Addition of biochar to soil has also been associated with increased nutrient
use efficiency, either through nutrients contained in biochar or through physico-chemical
processes that allow better utilisation of soil-inherent or fertiliser-derived nutrients.
Importantly, it is the apparent biological and chemical stability that allows biochar to both act
as a carbon sink, as well as provide benefits to soil that are long-lived.
Using pyrolysis to turn sustainably produced biomass into a recalcitrant substance that is
decomposed at a much slower rate, constitutes both a tool for carbon sequestration and
avoided emission. It is argued that sequestration of carbon in biochar allows for a much
longer storage time compared with other terrestrial sequestration strategies, such as
afforestation (Schulze et al., 2000). The stability and carbon sequestration potential of
biochar in soil is examined in Section 3.1.2.

2.1.1. Why and how is biochar made?

Modern industrial bioenergy systems involve pyrolysis and gasification, the heating of a
biomass feedstock under controlled conditions to produce combustible synthesis gas
(‘syngas’), and oil (‘bio-oil’) that can be burnt to produce heat, power, or combined heat and
power. Biochar, the third combustible product produced in pyrolysis, is the solid charred and
carbon-rich residue.
The balance in energy release and biochar formation can be optimised. Effectively, it is a
‘combustion’ process that may be curtailed at a point where any desired ratio in these
products has been achieved. This ratio can then be adjusted and re-optimised to satisfy
changing objectives. Whereas simple combustion of a feedstock maximises energy yield per
unit mass, combusting syngas from pyrolysis gives – where optimised for biochar – a much
greater energy yield per unit of carbon release.

3
If incorporating biochar into soil can reliably deliver the environmental benefits outlined in
3.1.2, the carbon-equivalent saving resulting from conversion of biomass using pyrolysis can
be increased further, relative to the production of energy only (i.e. combustion).
Globally, there is an estimated 15x109 ha of cropland (Ramankutty et al., 2008). On the basis
of calculations by Gaunt (2008), the application of biochar once every ten years to this land
area would result in a CO2 -equivalent gain of 0.65 GtC yr-1.

FEEDSTOCKS PROCESS PRODUCT USES and APPLICATIONS

Biomass energy Fast pyrolysis Synthesis gas - Heat

crops (corn, (anhydrous) Bio-oil liquid - Fuel (combusted to generate electricity
cereals, wood Biochar solid or converted to syngas)
pellets, palm oil, - High value biochemicals used as food
oilseed rape) Slow pyrolysis additives or pharmaceuticals
(low temp. 450-
450- Syngas
- Soil conditioners / fertilisers
550°C, O2-free,
-free,
Bioenergy residues sometimes
some-times steam)
steam)
“cake” Biochar - Soil amendment (neutral / alkaline pH,
porosity retains water, cation exchange
Slow pyrolysis Activated capacity: robust benefits to plant growth
Agricultural waste (high temp.600-
(wheat straw, Biochar compared to high-temp
high-temp char)
char)
900°C, O2-free) --Fuel
Fuel(cooking
(cookingand
andheat)
heat)
hazelnut and
peanut shells, Combustible
waste wood, etc) Gasification (high - Extreme porosity and surface area
ethane,
temp., fast heating methane - Water filtration and adsorption of
rate.,OO2 present)
rate, contaminants (gas, liquid or solid)
Compost (green
waste) Biochar
- Fuel (low yield, high reactivity)
Fermentation, - Contamination of some feedstocks
Manure/ animal
anaerobic digestion ethanol
Ethanol (e.g. metal and plastic in kitchen waste)
waste (chicken)
and mechanical may preclude use of sludge / char in soil
bio-treatment
Kitchen waste Methane and
sludge - Fuel (for electricity or cooking)
plastic, food, etc
Carbonisation - Bi-products
By-products(wood
(woodspirits,
spirits,wood
woodtar)
tar)
(‘brown’ at 300°C, - Substitute for coal
coal-derived
-derived coke
cokeinin
Sewage sludge ‘black’ at 380°C) Charcoal metal smelting

Figure 2. Summary of pyrolysis processes in relation to their common feedstocks, typical

products, and the applications and uses of these products

Pyrolysis has a requirement for initial energy, in the same way as in straight combustion
some heat in the flame is used to initiate combustion of new feedstock. But the relative
requirements must be carefully compared, together with any difference between pyrolysis
and alternative bioenergy technologies in the energy requirement of feedstock transportation
and drying.
The potential advantage of pyrolysis-derived bioenergy over other bioenergy strategies in
terms of greenhouse gas emissions results not only solely from the retention of up to 50% of
the feedstock carbon in stable biochar, but from indirect savings that may result from the use
of biochar in agriculture, specifically the soil (Gaunt et al., 2008).
Biomass pyrolysis and gasification are well established technologies for the production of
biofuels and syngas. However, commercial exploitation of biochar by-products as a soil
amendments is still in its infancy. In Japan, which has the largest market for such products,
approximately 15 000 t yr-1 is traded annually for soil use (Okimori et al., 2003). More usually
biochar products are gasified for extraction of residual energy, or used in production of high
value products such as activated carbon (Demirbas et al., 2006b).
The pyrolysis process greatly affects the qualities of biochar and its potential value to
agriculture in terms of agronomic performance or in carbon sequestration. The process and
process parameters, principally temperature and furnace residence time, are particularly
important; however, the process and process conditions also interact with feedstock type in
determining the nature of the product.

4
These variables together are key influences on the chemical, biological and physical
properties, which limit the potential use for biochar products. A summary of feedstock,
production processes and products is illustrated in Figure 2. The assignment of feedstock to
process in Figure 2 is based on examples from the literature and does not indicate that they
should be an exclusive classification.
Each category of pyrolysis process is characterised by a contrasting balance between
biochar, bio-oil and syngas (Table 1). The precise ratio in these products may vary between
plants, and may be optimised at a particular installation; however, it is critical that maximising
the production of biochar relative to mass of initial feedstock (Demirbas, 2006), is always at
the expense of usable energy in the liquid or gaseous form. Although a greenhouse gas
mitigation strategy may favour maximising the biochar product (Gaunt et al., 2008), the
balance that is realised is a function of market and engineering constraints.
In a generalised analysis, the economic cost of maximising the retention of carbon in biochar
using slow pyrolysis has been assessed against the possible net gain in CO2 –equivalent
emissions from using the product in soil, after accounting for the additional fossil-carbon
offset that could be obtained through complete combustion of the feedstock (Gaunt et al.,
2008). The net carbon gain over fossil fuel scenarios was 2–19 t CO2 ha-1 y-1, encompassing
figures 2–5 times higher than those for strategies based on biomass combustion. The eligible
portion of this added saving would have to attract CO2 -offset at a value sufficient to cover
the USD 47 t-1 value of residual energy in biochar. A more detailed description of this
analysis and discussion of the competing processes associated with energy and char
production is discussed in detail in Chapter 3.

Table 1. Fate of initial feedstock mass between products of pyrolysis processes (IEA, 2007)

Process Liquid Solid Gas

(bio-oil) (biochar) (syngas)

FAST PYROLYSIS
75%
Moderate temperature (~500 °C) 12% 13%
(25% water)
Short hot vapour residence time (<2s)

INTERMEDIATE PYROLYSIS
50%
Low-moderate temperature, 25% 25%
(50% water)
Moderate hot vapour residence time

SLOW PYROLYSIS
30%
Low-moderate temperature, 35% 35%
(70% water)
Long residence time

GASIFICATION
5% tar
high temperature (>800 °C) 10% 85%
5% water
Long vapour residence time

2.1.2. Biochar feedstocks

Although current results suggest that the type of feedstock used for pyrolysis is more
important where biochar is to be applied as a soil conditioner (Section 3.2.) there is little
consensus as to what constitutes optimal feedstock for energy production.
This is mainly due to the fact that the number of existing commercial plants is small, and that
these plants are dedicated to specific waste streams, giving little incentive to experiment with
this parameter. However, some research-scale pyrolysis plants have conducted experiments
with a wider range of feedstocks (Day et al., 2005; Das et al., 2008; Gaunt et al., 2008).
Feedstocks currently used at a commercial-scale or in research facilities include wood chip
and wood pellets, tree bark, crop residues (including straw, nut shells and rice hulls), switch
grass, organic wastes including distillers grain, bagasse from the sugarcane industry and

5
olive waste (Yaman, 2004), chicken litter (Das et al., 2008), dairy manure, sewage sludge
(Shinogi et al., 2002) and paper sludge.
The elemental ratios of carbon, oxygen and hydrogen are key feedstock parameters in
commercial use and the quality of fuel products (Friedl et al., 2005). The feedstocks which
are favoured for bio-oil and fuel-gas are those that have low mineral and N content. These
include wood and biomass from energy crops, including short-rotation woody plants (such as
willow), high productivity grasses (such as Miscanthus spp.), and a range of other
herbaceous plants. They may also include abundant, available and low-cost agricultural by-
products, including cereal straw.
The proportions of hemi-cellulose, cellulose and lignin content determine the ratios of volatile
carbon (in bio-oil and gas) and stabilised carbon (biochar) in pyrolysis products. Feedstocks
with high lignin content produce the highest biochar yields when pyrolysed at moderate
temperatures (approx. 500 °C) (Fushimi et al., 2003; Demirbas, 2006). In the future, selection
of feedstock may be dictated by the desired balance between pyrolysis products (gas, oil and
biochar), and whether the production process is slow pyrolysis, or a related process.
Charring of agricultural waste products such as nut shells and rice hulls for energy
production may be advantageous compared to disposal as waste by some other means
(Demirbas, 2006; Demirbas et al., 2006a). Alternative use for such materials includes
composting and mulching. However, it is important to recognise that continuous removal of
crop residues from the same land compromises soil cover and diminishes soil nutrient
supply. This is further discussed in Chapter 3.

2.1.3. Biochar production systems

Biochar is a multi-process product whose qualities are dependent on each process and also
the material to which the process is applied. Since the technology is still in a period of
development and not yet optimised to producing a product for use in soil, it is useful to review
the various technologies currently in use. The processes of slow- and fast-pyrolysis are
exemplified in Figure 3 and 4 and discussed in the subsequent sections.

6
Figure 3. Illustrative slow pyrolysis process (Source: BEST Energies)

Char
separation
Gas

Pyrolysis
Fluid bed
Drying Grinding CFB Bio-oil
Biomass to < 10% water to < 3mm Transported bed
Rotating cone cool and
Entrained flow collect
Ablative
Etc. Char

Figure 4. Schematic process diagram for fast pyrolysis (modified from Bridgewater (2006))

2.1.3.1. Slow pyrolysis

Slow pyrolysis is the thermal conversion of biomass by slow heating at low to medium
temperatures (450 to 650°C) in the absence of oxygen, with the simultaneous capture of
syngas. Feedstocks in the form of dried biomass pellets or chips of various particle sizes are
fed into a heated furnace and exposed to uniform heating, generally through the use of
internal or external heating as retort furnace or kilns, respectively. Conventional production
conditions have been summarised by Peacocke and Joseph in <http://www.biochar-
international.org/images/Terminology.doc> :
• residence times: >5 seconds for the production of syngas; minutes, hours or days for
biochar production
• relatively low reactor temperatures (450-650°C)
• reactor operating at atmospheric pressure
• very low heating rates, ranging from 0.01–2.0°C s-1
• very short thermal quenching rate for pyrolysis products: minutes to hours.

7
Several commercial facilities generate syngas and biochar using a continuous flow system in
which feedstock passes slowly through a kiln in an auger feed, with combustible syngas
continuously drawn away.
Biochar, bio-oil and syngas are formed in approximately equal proportions due to the slow
speed of the combustion process, which promotes extensive secondary reactions within
biochar particles and in the gas and vapour phases, leading to condensation.
The pyrolysis reaction itself is mildly endothermic, with the bulk of energy capture being in
the form of the syngas and bio-oil condensates. The biochar has a residual energy content of
about 30–35MJ kg-1 (Ryu, 2007), and conventionally this is extracted within the plant by
burning or gasification, providing heat to drive the primary pyrolysis (Demirbas, 2006), or to
dry incoming feedstocks. The syngas product may be combusted on site to generate heat or
electricity (via gas or steam turbine), or both.
A variant of slow pyrolysis includes a steam gasification step, a technology developed by
Eprida in collaboration with the University of Georgia (<http://www.eprida.com/hydro/>).
Adding steam to the pyrolysis reaction liberates additional syngas from the biochar product,
mainly in the form of hydrogen. The biochar that remains after this ‘secondary’ pyrolysis
displays rather different properties from the primary product, differing in pore size and carbon
to oxygen ratio (Demirbas, 2004).
Syngas can be purified through a sequence of operations to yield pure streams of the
constituent gases: hydrogen (50% of gas yield), carbon dioxide (30%), nitrogen (15%),
methane (5%), and lower molecular weight hydrocarbons, as well as some carbon monoxide
(Day et al., 2005). There is a small energy penalty associated with these steps.
Slow pyrolysis research plants currently process feedstock at a rate of 28–300 kg hr-1 on a
dry mass basis, and commercial plants operate at 48–96 t d-1. Comparing the efficiency of
pyrolysis plants is complex since the mix and use of products vary, and the composition and
heat value of syngas differs. Feedstock quality and moisture content is also variable, and
there is a conversion loss in the generation of electrical power through steam or gas turbines.

2.1.3.2. Fast pyrolysis

Very rapid feedstock heating leads to a much greater proportion of bio-oil and less biochar
(Table 1). It was with the objective of achieving this high yield of liquid fuel that fast pyrolysis
technology was developed.
The time taken to reach peak temperature of the endothermic process (the ‘resistance time’)
is approximately one or two seconds, rather than minutes or hours as is the case with slow
pyrolysis. The lower operating temperature also enhances the overall conversion efficiency
of the process relative to slow pyrolysis.
Maintaining a low feedstock moisture content of around 10% and using a fine particle size of
<2mm permits rapid transference of energy despite relatively moderate peak temperatures of
around 450°C (and in the range 350 to 500°C). In many systems the transfer is further
increased by mechanically enhancing feedstock contact with the heat source or maximising
heat source surface area. Various technologies have been used and proposed or tested
including: fixed beds, augers, ablative methods, rotating cones, fluidized beds and circulating
fluidized beds. Surface charring must be continuously removed during reaction to prevent
pyrolysis of particle interiors being inhibited by its insulating effect.
Bio-oil is condensed from the syngas stream under rapid cooling, with the combustion of
syngas providing the pyrolysis process heat. The bio-oil is a low grade product with a calorific
value, on a volume basis, approximately 55% that of regular diesel fuel. It is unsuitable as a
mainstream liquid transport fuel even after refining, and is most suitable as a fuel-oil
substitute. It is considered to have an advantage over typical fuel oils in zero SOx and low
NOx emission on combustion (Bridgewater, 2004). In addition to combustion for electricity
generation, bio-oil may be converted to syngas for production of clean fuels (gasification).

8
Bio-oil also contains high value bio-chemicals of relevance to food and pharmaceutical
industries.
The biochar product of fast pyrolysis is granular and displays a lower calorific value (23–
32 MJ kg-1) than that of slow pyrolysis (Demirbas, 2001). However, there are currently no
published studies to assess the effects of biochar from fast pyrolysis when it is applied to
soil. It is highly likely that condensed volatiles will be present in the product and that this will
affect its performance and desirability.

2.1.3.3. Intermediate pyrolysis

This term describes a hybrid technology under evaluation at Aston University, UK, designed
to produce bio-oil with very low tar content, with perceived potential for use as a motor fuel.
The process has been tested with woody and non-woody feedstocks, and produces biochar
in greater quantity and of contrasting quality as compared to fast pyrolysis.

2.1.3.4. Carbonisation

Carbonisation describes a number of pyrolysis processes that most closely resemble

traditional, basic methods of charcoal manufacture, and which produce biochar of the highest
carbon content. The auto-thermal carbonisation process is the small-scale method widely
used in rural communities around the world (FAO, 1987). The second requires fossil fuel to
provide an external heat source, and is associated with industrial, mass production of
charcoal (FAO, 1985). The process is optimised for the solid products of pyrolysis, but
condensed gases provide an industrial product known as ‘wood vinegar’, which as well as
providing the basis for food flavouring ingredients, is considered to have a fertiliser value to
plants.
The auto-thermal process as the most realistic option has been proposed for the participation
of local communities in using biochar to build soil fertility, especially in developing countries.
It is lower cost, and easier and simpler than pyrolysis systems where ratios of solid, liquid
and syngas products have to be optimised. A comparison of three alternative options has
recently been investigated for the carbonisation of biomass wastes from tree plantations: a
drum kiln, a Hume pipe kiln, and a brick kiln (Okimori et al., 2003; Ogawa et al., 2006). For
wood, 24% of wood mass was converted to biochar of 76% carbon content at 400 to 500°C,
but carbonisation at 600°C gave 28% biochar with a higher carbon content of 89%. The 50%
of feedstock carbon stabilised in each case was similar to the maximum yield obtained in
slow pyrolysis.
Brazil has the largest concentration of industrial charcoal manufacture. This is associated
with the pig-iron (smelting) industry, where substitution of charcoal for fossil-derived coke has
been achieved in a number of very large projects under the Clean Development Mechanism,
and associated with large-scale plantations of eucalypt.

2.1.3.5. Gasification

Gasification is the process by which any carbonaceous material (coal and petroleum as well
as biomass) is substantially converted into a stream of carbon monoxide and hydrogen in a
high temperature reaction and controlled-oxygen environment, sometimes at high pressures
of 15–50 bars (Bridgewater, 2006). The gas mixture is the key energy output and the
gasification process has an application as a clean waste disposal technique (Bapat et al.,
1998). In slow pyrolysis facilities, gasification is often used to generate further syngas from
biochar ‘waste’.
Syngas may be used for electricity generation via gas or steam turbines (or both), used to
manufacture chemicals and fertilisers, or further cleaned for use as liquid fuel. Since
conversion of feedstock to syngas is often the main objective, the process is maximised for
gas production and so the biochar yield from gasification tends to be very low (Table 1).
However, this also carries the risk of higher levels of metals and minerals that may be

9
concentrated in biochar, with potential safety implications with regard to application to soil
(Fernandes et al., 2003a). Although biochar from gasification has a high calorific value
(Demirbas, 2006; Demirbas et al., 2006a), it has high value use as the basis for activated
carbon production. Its potential to act as the carrier in a slow nutrient release fertiliser
product has also been noted (Ueno, 2007).
Worldwide, gasification has been used on a commercial scale worldwide for more than 50
years, mainly in the refining, fertiliser, and chemical industries, and for more than 35 years in
the electric power industry. There are more than 140 gasification plants currently in
operation, with worldwide gasification capacity projected to grow 70% by 2015, with 80% of
this growth occurring in Asia (Anon, 2008). The majority of gasification plants produce
chemicals, mainly methanol or ammonia and urea, and use coal or refuse-derived feedstocks
(Kedco, 2008). However, biomass feedstocks are also used, for example wood pellets, wood
chips or paper and sugar cane bagasse (Ueno et al., 2007), and rural gasification projects
are expanding.

2.1.3.6. Production of ammonia during pyrolysis

In the Haber-Bosch process, a fossil hydrogen source (usually methane) is used to fix
atmospheric nitrogen and create ammonia for manufacture of fertiliser. Hydrogen in syngas
streams from pyrolysis (7-8% of slow pyrolysis syngas) can substitute methane and be used,
potentially not only to create ammonia but, if conducted on the same site as the pyrolysis, fix
ammonia to a biochar co-product. This offers the prospect of a crop fertiliser that
simultaneously adds stabilised carbon to soil, possibly with slow release characteristics. This
could offer greater net benefit in terms of CO2 -equivalents than using syngas to, for
example, generate electricity. The process of fixing ammonia from the hydrogen syngas
stream at atmospheric pressure and ambient temperature has been demonstrated (Day et
al., 2005) and is illustrated diagrammatically in Figure 5. However, agronomic evaluation of
the product has not been published and the concept has not yet been commercialised.

Figure 5. Simplified flow chart of how biomass releases energy as it captures CO2 as
ammonium carbonate (modified from Day et al., 2005 )

2.2. Biogeochemical characterisation of biochar

2.2.1. The purpose of biogeochemical characterisation

Although commercial biochar products are being developed for use in soil, credit for carbon
storage will require predictable levels of stability and the ability to verify actual rates of
degradation through quantitative soil analysis (Ogawa et al., 2006; Matthews, 2008).
Identifying the characteristics that determine the stability of biochar will enable its properties
to be optimised and standardised in production. Techniques to unambiguously detect and

10
measure its abundance in soil are necessary to verify its application and to trace the fate of
particular biochar products.
Biogeochemical characterisation techniques may also help understand the function of
biochar products in soil at the process level. With specific knowledge of the nature of
biochar-driven processes in the soil, predictive capacity for the longevity and reactions of
biochar in soil that will determine its value as a soil carbon sink and soil conditioner are
possible.
From a practical point of view it is important that the methods devised enable biochar
characteristics to be determined sufficiently rapidly and inexpensively to permit widespread
application and use.

2.2.2. Physical and chemical characterisation

Kuwagaki (1990) proposed that seven properties should be measured for a quality
assessment for agronomically-used biochar: pH, volatile compound content, ash content,
water holding capacity, bulk density, pore volume, and specific surface area.
Feedstock is a primary factor governing the chemical and physical properties of biochar. The
elemental composition reported for a range of bio-oil and biochar products from various
feedstocks are compared in Table 2.

Table 2. Reported elemental composition for a range of bio-oil and biochar products (% dry
ash-free mass)
Elemental composition (%) HHV*
Product C H N O (MJ/kg)

Beech-trunk bark biochar 87.9 2.9 0.6 10.6 33.2

Beech-trunk bark bio-oil 68.8 8.9 0.8 21.5 34.6
Rapeseed cake biochar 66.6 2.5 6.1 24.3 30.7
Rapeseed cake bio-oil 73.9 10.8 4.7 10.6 36.5
Wood bark biochar 85.0 2.8 – 12.2 30.8
Wood bark bio-oil 64.0 7.6 – 28.4 31.0
Cotton stalk biochar 72.2 1.2 – 26.6 21.4
Cotton stalk bio-oil 59.7 7.8 1.8 30.6 26.0
Bio-char from hazelnut shell 95.6 1.3 – 3.1 32.0
Sunflower bio-oil 72.1 9.8 5.2 12.9 36.2

*HHV= higher heating value (enthalpy of complete combustion of a fuel including the
condensation enthalpy of former water); Demirbas et al.,(2004)

In general, the carbon content of biochar is inversely related to biochar yield. Increasing
pyrolysis temperature from 300 to 800°C decreased the yield of biochar from 67 to 26% and
increased the carbon content from 56 to 93% (Tanaka, 1963). Beyond a certain threshold,
the mass of biochar may decrease without any affect on the amount of carbon retained within
it; but as mass is lost, the ash content of biochar increases. In one study, the proportion of
biochar comprised of ash increased from 0.67 to 1.26% between 300°C and 800°C
(Kuwagaki and Tamura, 1990).
For a particular feedstock, the elemental composition of the pyrolysis products can still be
greatly affected by the processing temperature and pyrolysis residence time. The effect of
temperature on the composition of biochar from sugarcane bagasse is listed in Table 3.

11
There was a corresponding impact on the pH of the biochar from 7.6 at 310°C to 9.7 at
850°C (Kuwagaki and Tamura, 1990).

Table 3. Properties of biochar from bagasse carbonisation (Ueno et al., 2007)

Parameter or property Biochar Feedstock

o
Preset temperature ( C) 500 600 700 800
Average temperature (oC) 490 690 740 830
Specific surface area (m2/g) Nd 270 322 273 Nd
-1
EC (mS m ) 7.78 7.15 6.95 7.83 Nd
pH 7.46 7.59 7.68 7.89 Nd
TN (%) 0.58 0.45 0.32 0.44 0.19
TC (%) 70.5 71.0 65.2 73.9 46.1
-1
Minerals (mg 100g ) 3361 4601 5359 4363 841

nd=not determined

There are inevitable accompanying differences in the physical and other chemical properties
of biochar. Scanning electron microscopy (SEM) is often used to describe the physical
structure of biochar, and the architecture of cellulosic plant material is clearly retained (Figure
6). It has been suggested that the porous structure of biochar can explain its impact on soil
water holding and adsorption capacity (Day et al., 2005; Ogawa et al., 2006; Yu et al., 2006).

Figure 6. Scanning electron microscope (SEM) image of biochar produced at 400°C from
pelletised peanut shell (Jason Nadler, Eprida, Day et al., 2005)

12
Process temperature greatly affects the surface area of pyrolysis products. In one study,
surface area was shown to increase from 120 m2g-1 at 400°C to 460 m2g-1 at 900°C (Day et
al., 2005). This effect of temperature has led to suggestions that biochar created at low
temperature may be suitable for controlling the release of fertiliser nutrients (Day et al.,
2005), whilst high temperature biochars would be more suitable for use as activated carbon
(Ogawa et al., 2006). The surfaces of low temperature biochar are, however, hydrophobic
and this may limit the capacity to store water in soil.
The form and size of the feedstock and pyrolysis product may affect the quality and potential
uses for the biochar product. Initially, the ratio of exposed to total-surface-area of biochar is
affected by its particle size. However, although low temperature biochar is stronger than high
temperature products, it is brittle and prone to abrade into fine fractions once incorporated.
Thus, over the long term, surface area, i.e. of weathered biochar, may not be greatly affected
by this parameter.
Biochar comprises part of a continuum of materials described as ‘black carbon’ (Schmidt et
al., 2001), which are difficult to quantify. Techniques that have been used to characterise this
wider class of materials – which includes soot, charcoal, and char from vegetation fire – may
be applied to detect the presence of biochar in soil, sediments and air.
These methods include extractive techniques analogous to those applied to soil organic
matter, solid state 13C nuclear magnetic resonance (NMR) spectroscopy (with cross-
polarization, CP, or Bloch decay, combined with magic angle spinning, MAS), diffuse
reflectance infrared Fourier transform spectroscopy (DRIFT), and near-edge X-ray
absorption fine structure (NEXAFS) spectroscopy (Baldock et al., 2002; Fernandes et al.,
2003b; Lehmann et al., 2005b).
Biochar occurs naturally in soil as a consequence of wildfire (see Section 3.1.5.1.) and is
considered to be a distinct soil carbon pool (Krull et al., 2003). Research to quantify and
characterise this material has highlighted the limitations of various analytical techniques
when applied to soil and separation procedures to physically separate char from other soil
organic matter. A key problem is the interference that the mineral matrix poses, particularly
with its associations with highly stabilised soil organic matter, which chemically resembles
char. This has been successfully addressed through the use of high energy ultraviolet photo-
oxidation, which oxidises most non-char carbon (Smernik and Oades, 2000; Smernik et al.,
2000). Thermal oxidation to remove lignin (De la Rosa et al., 2008) and chemical oxidation
using hydrofluoric acid to remove mineral interferences (Simpson and Hatcher, 2004a) have
also been used.

2.2.3. Quantification of biochar and char

The categorisation of organic carbon in soil is in general a major challenge. Quantifying

different components of black carbon is particularly difficult, on account of its chemical
complexity and its inherently un-reactive nature. To compare the efficacy of candidate
methods depicted in Figure 7, a ring trial was recently undertaken using a selection of 12
materials of contrasting origin and source (Hammes et al., 2007).
Methods included in the trial included approaches based on biomarkers, particularly benzene
polycarboxylic acids (BPCA). Chemical oxidative treatments using acid dichromate or sodium
chlorite were represented; other oxidative approaches used ultra-violet treatment (photo-
oxidation) with correction for non-black aromatic carbon through 13C NMR analysis of
oxidised residues. A chemo-thermal oxidation method used a temperature threshold of
375°C, also with 13C NMR and elemental analysis of residues. A purely thermal (gravimetric)
analysis was represented using a helium gas flow containing 20% oxygen. Purely
spectroscopic methods included thermal or optical laser transmittance and reflectance, and
mid-infrared. Additional methods assessed were matrix assisted laser desorption ionisation
(MALDI –TOF) and hydrogen pyrolysis.
The most promising reference techniques for determination of biochar in soils were UV or
chemical oxidation with elemental analysis and 13C NMR analysis of residues (Skjemstad et

13
al., 1999; Smernik et al., 2002), thermal analysis (De la Rosa et al., 2008; Hammes et al.,
2007) and hydrogen pyrolysis (Ascough et al., 2008).
Mid-infrared spectroscopy shows promise as a means to rapidly estimate black carbon (Janik
et al., 2007) and has been applied to large sample sets (Lehmann et al., 2008). This is a
correlative technique calibrated against a reference method.

S lig h tly
c h a rre d Char C h a rc o a l Soot GBC
b io m a s s

m m a n d la rg e r m m to s u b m ic ro n
p a rtic le s s u b m ic ro n

h ig h ly in e rt
re a c tiv e

U V O X ID A T IO N – 13C NM R

u ltra -h ig h re s o lu tio n
m a s s s p e c tro m e try

th e rm o -c h e m ic a l
v is u a l / m e th o d s
m ic ro s c o p ic
c h e m ic al
m e th o d s

BCPAs

o th e r m o le c u la r
m a rk e rs

Figure 7. Components of black carbon assessed by available quantification methods across

a continuum of recalcitrance increasing from left to right (Hammes et al., 2007)

2.2.4. Background and biochar history: from terra preta to terra nova

The terra preta of the Brazilian Amazon are anthropogenic dark earths, characterised by
enhanced levels of soil fertility and popular locally for growing cash crops such as papaya
and mango. These crops are said to grow three times faster than on surrounding land, a
landscape characterised by soils of generally low fertility. Although the terra preta occur in
small patches averaging 20 ha, sites as large as 350 ha have been reported (Smith, 1999).
Similar soils have not only been identified elsewhere within the region, namely Ecuador and
Peru, but also beyond, in West Africa (Benin, Liberia), and the savanna of South Africa
(Lehmann et al., 2003).
The terra preta display high levels of soil organic matter (SOM) and nutrients such as
nitrogen, phosphorus, potassium and calcium (Table 4). These characteristics and their high
fertility is attributed in part to a high char content (Glaser et al., 2001), which is the main
reason why the terra preta tend to be much darker in colour than adjacent soils (Figure 8).
The source of the large amounts of char is considered to have been incompletely combusted
biomass carbon, such as wood from kitchen fires or in-field burning, conducted by relatively
large and settled communities in the distant past.

14
The terra preta phenomenon has widespread public appeal and has attracted extensive
coverage in popular science publications, TV and film, and a plethora of websites. The
potential relevance of the terra preta as a model for modern day variants using by-products
of bioenergy is now well established and recognised in popular science (Lehmann, 2007b;
Baskin, 2006) as well as high impact scientific journals (Marris, 2006; Lehmann, 2007a).
From this, the idea of terra nova has emerged: soils whose properties would be enhanced by
modern variants of the management practices that created terra preta,

Figure 8. Comparison of profiles of terra preta and adjacent soils (Source: IBI website)

Table 4. Physico-chemical properties of terra preta and adjacent soils (data from Solomon et
al., 2007; Liang et al., 2006)

Total
Organic soil
Depth Clay carbon nitrogen
range content (mg g-1 (mg g-1 C:N
Site Soil type (cm) Age (yr) (% soil) pH soil) soil) ratio

terra preta 43-69 600-1000 27.0 6.4 22.0 1.0 23

Hatahara
Adjacent soil 0-10 600-1000 35.9 4.6 21.8 1.6 14

terra preta 0-16 900-1100 22.6 5.9 31.5 1.8 18

Lago
Grande
Adjacent soil 0-8 900-1100 26.7 4.2 17.5 1.3 14

terra preta 48-83 2000-3000 10.4 5.6 15.7 1.0 16

Acutuba
Adjacent soil 0-30 2000-3000 8.5 4.7 15.4 0.8 20

terra preta 190-210 6700-8700 0.3 5.0 16.5 1.1 15

Dona
Stella
Adjacent soil 0-12 6700-8701 0.3 3.9 10.2 0.4 27

15
3. BIOCHAR APPLICATION IN AGRICULTURE
In the context of this report, carbon sequestration is the primary driver for considering the
application of biochar to soil. Policy makers charged with meeting greenhouse gas emission
targets and addressing public concern over increasingly evident climate change may
recognise the potential for biochar-based strategies. The land-owner or farmer is likely to
have a more practical or financial perspective.
A particular combination of feedstock, pyrolysis technology, energy conversion and by-
product usage can comprise a biochar-based system. Alternative systems have different
greenhouse gas balances. In Section 3.1.4, economically and politically conceivable systems
for different regions of the world are considered. The future price of carbon and the inclusion
of biochar in carbon-trading schemes is a key factor. The likely additional benefits of biochar
to agricultural production are reviewed in Section 3.2. These factors are critical since they
dictate whether relevant practices are adopted on a large scale through their effect on the
decision making of individual farmers.
From a global and policy perspective the potentially negative impacts of biomass use on
climate forcing must be considered. These include the effects of soot and trace gases that
are emitted into the atmosphere during combustion. Airborne transport and deposition of soot
has been implicated in the acceleration of polar ice melt, but conversely in facilitating cloud
formation and ‘global dimming’ (McConnell et al., 2007; Ramanathan et al., 2008).
Currently biomass burning accounts for 10% of global CH4 emissions and 1% of N2O
(Crutzen et al., 1990). Although current charcoal production activity could account for a
component of these emissions (Woolf, 2008), a general shift to pyrolysis-based systems
would decrease, if not eliminate, them. However, the net result with great expansion of
alternative bioenergy systems has not been assessed.
As is apparent in the terra preta, the addition of biochar can dramatically darken the colour of
soil, especially in soils that are low in organic matter. A relationship between soil colour and
low temperature fire occurrence has been demonstrated (Ketterings et al., 2000). Oguntunde
(2008) found soil at charcoal manufacturing sites to have 8% greater hue, and 20% higher
value and chroma. Since dark soils absorb more solar energy they may, depending on water
content and plant cover, display higher soil temperatures (Krull et al., 2004). This will affect
rate processes, enhancing the cycling of nutrients and potentially extending the growing
season in seasonal climates. In Japan it is a traditional farming practice to apply charcoal to
accelerate snow melt. Anecdotal evidence suggests more rapid crop establishment in
temperate soils enriched in char, but to date no quantitative relationships between biochar
application rate and these parameters have been reported. The study of Oguntunde (2008)
showed a one-third reduction in soil albedo in soils enriched in char. On a large spatial scale,
the application of biochar could affect the albedo of the Earth’s surface. Increasing surface
albedo has been proposed as a possible mitigation measure for climate forcing (Crutzen,
2006).
The frequency with which potentially toxic compounds materialise in biochar and their
concentration is inadequately researched. Two classes of compounds are of generic
concern, since they can potentially from in the pyrolysis of any feedstock: polycyclic aromatic
hydrocarbons (PAH) and dioxins. These compounds and available evidence for their
presence in biochar is examined in Section 3.2.6.

3.1. Biochar and climate change

The natural carbon cycle includes natural char production from wildfires, the ensuing
transport of char from the soil to watercourses and the ultimate burial in marine or estuarine
sediments. Since routine and universally acceptable methods for black carbon and
specifically charcoal are still outstanding, the precise magnitude of the rates and processes,
and the relative size and stability of char in the soil and sediment pools are still uncertain

16
(Schmidt, 2004; Simpson and Hatcher, 2004b). However, the potential to enhance the
contribution that char makes to the natural carbon cycle through the addition of biochar in soil
is a topic of much public discussion and a rising profile in influential policy circles, for
example, in Australia (Garnaut, 2008). The contribution that such strategies can have on
climate change mitigation depends on attaining a much more extensive research base and
detailed economic analyses.
It is useful to consider a biochar-based strategy against more established approaches to
increase the organic carbon stored in soil, such as the use of manures and composts. The
longevity of biochar in the soil is an important element when comparing pyrolysis bioenergy
and biochar production with conventional bioenergy strategies, in mitigating climate change.
However, it is also vital to assess any indirect reduction in net greenhouse gas emissions
from agriculture through the use of biochar. There may be additional benefits arising from the
contribution of biochar to facilitating agricultural development and improving the socio-
economic circumstances of farmers in developing countries. Figure 9 captures the
complexity of potentially beneficial interactions of biochar in the context of natural cycles and
anthropogenic interventions.

3.1.1. Soil organic matter and climate change

In order to understand the potential significance of carbon in soil in the form of biochar, its
characteristics and dynamics should be compared to those of the remaining soil organic
matter, which accounts for most of the carbon that exists in soil (the exception being
calcareous soils which contain stocks of inorganic carbon in carbonate minerals). Depending
on land-use and climate, most soils contain up to approximately 100 t ha-1 carbon as organic
matter. Peat soils, though, comprise mainly organic matter and contain much more carbon
on a per unit area basis. It is increasingly recognised, however, that a greater proportion of
the total carbon may comprise an accumulated store of the products of burning or fire
(Skjemstad et al., 2004a), and that this has implications for the response of the wider soil
carbon pool to climate change (Skjemstad et al., 1999; Lehmann et al., 2008).
Modelling indicates that about 90% of the organic matter present in soils turns over on
decadal to centennial timescales (Coleman et al., 1996; McGill, 1996). Most organic matter in
soil is derived from plant roots, plant debris and microbially re-worked substances. The
presence of soil organic matter is important for a range of useful soil properties, which has
been comprehensively reviewed by Krull (2004). The process of microbial energy acquisition
(and concomitant CO2 release) from substrate is accompanied by a release of various
nutrient elements, which may be conserved in the soil in microbial biomass or the particulate
residues of substrate decomposition. A portion of certain nutrients may also be released in
soluble form, and a fraction may be lost from the soil through leaching or run-off; which is
essential to crop nutrition. This is particularly the case where external nutrient provision (from
fertiliser or manure) is limited or absent.
Overall, a balance slowly develops between the rate of carbon addition and the emission of
CO2, which are specific to the land-use and environmental conditions. The amount of organic
matter maintained once this balance is reached, depends on its average rate of turnover. To
date there have been few means proposed that permit manipulation of this rate, so that soil
carbon can be permanently increased. Beyond simply increasing the amount of external
organic matter inputs (Smith et al., 2000), the main strategies are to disturb the soil less by
using less intensive tillage or zero tillage (Lal, 1997; Smith et al., 1998), or by selecting
particularly recalcitrant, lignin-rich amendments (Palm et al., 2001).
Although conversion to no-till soil management has been widely promoted as an approach to
enhance soil organic matter as well as to control erosion and conserve water, the main effect
appears to be a vertical re-distribution of organic matter, and an increase toward the surface
more or less matched by a corresponding depletion at depth (Bhogal et al., 2007; Blanco-
Canqui et al., 2008). Nonetheless, the Chicago Climate Exchange includes a specification for
‘conservation tillage’ amongst its Carbon Financial Instruments for carbon sequestration and
thus a precedent for the active engagement of farming in the carbon market

17
(<http://www.chicagoclimatex.com/>). Managing decomposition in soil by manipulating the
quality of inputs has been explored extensively in tropical environments where decay is rapid
(Palm, 2001). But simply altering the composition of soil inputs has only a relatively minor
impact on the composition and long-term fate of the small portion that is stabilised, with
incorporation and repeated decomposition inside the dominant, slow turnover pool.
Thus the main emphasis in the sequestration debate has been focused on increasing soil
carbon by increasing organic matter additions in the form of straw or other crop residues, and
from external sources such as manures and a range of organic wastes: sewage sludge,
municipal compost, paper waste, and so on. Although there is a large amount of such
material available, the quantity is relatively small compared with the total flux through soil,
particularly the size of the global soil carbon pool. When a soil is at an equilibrium, only about
10% of the carbon added to soil is stabilised for more than one year. During a transition,
progress to new equilibrium is slow, with the annual increase being small relative to the
carbon invested. As equilibrium is approached the annual rate of accumulation decreases,
and once reached, the new level of input has to be sustained simply to maintain it.
Furthermore, the capacity to store organic matter is ultimately limited (with the capacity
varying with soil type, water regime and climatic factors); thus the improvement in carbon
storage that is possible for each incremental increase in input (Stewart et al., 2007; Gulde et
al., 2008). As well as added carbon being rapidly re-emitted into the atmosphere, carbon is
lost in the formation of soil organic matter through digestion in the animal gut or oxidation in
conversion to compost. The level of carbon sequestration or offset that could be realised
through an alternative use of these materials, including fossil fuel substitution, must be
considered when assessing the efficacy of these strategies from the perspective of climate
mitigation alone (Schlesinger, 2000). For example, the carbon cost of producing N fertiliser is
relevant when proposing to increase soil carbon storage indirectly through enhanced plant
growth (Schlesinger, 2000).

CO2

SOIL MANAGE PLANT

organic matter climate change biomass

CROP ENERGY
productivity capture

water N supply N2O effect

soot charcoal biochar

AGRICULTURAL
development

Figure 9. The key physical (purple arrows), natural (orange arrows) and anthropogenic (red
arrows) interactions of biochar in the environment

18
In the context of the interventions generically referred to as ‘management options’, important
soil physical benefits may be gained by accumulating soil organic matter (Janzen, 2006).
However, these must be balanced against the opportunity costs, the forgone benefits that
might arise from its breakdown and turnover, most importantly the release of crop nutrients
(Janzen, 2006). In general, however, any form of organic matter added to the soil degrades
resulting relatively quickly in CO2 emission. Thus adding degradable organic matter into the
soil is inefficient in terms of climate change mitigation, with the energy contained being
captured and dissipated by soil microbes rather than in power plants where it can offset fossil
fuel use (Woolf, 2008).

3.1.2. Carbon stabilisation and sequestration using biochar

Turning biologically-derived organic matter into a highly stabilised form can decrease CO2
emission from soil by considerably lowering its rate of decomposition. Whether stabilisation
by conversion to biochar represents a net carbon saving depends on the time horizon for the
comparison. However, the immediate CO2 emission from syngas released by pyrolysis
would, within a few months, be exceeded by the CO2 emitted in decomposition if the same
material had been added to soil directly (Lehmann et al., 2005a). Also, although there is a
CO2 emission associated with provision of heat for the pyrolysis process, the calculations of
Gaunt (2008) indicate that it is relatively small; in an example where pyrolysis consumed
40% of the carbon in the feedstock (in producing syngas), the CO2 resulting from provision of
process heat for that process would equate to only a further 10%. Even in temperate
environments where decomposition in soil is relatively slow, it is calculated that within two to
five years, the effective emission in the pyrolysis scenario is already less than that which
would have accrued from the soil (Gaunt et al., 2008). Over a period of the one to five
decades relevant to mitigation of climate change, the net saving is therefore considerable.
Even within the first few years, however, the higher initial loss of CO2 may be offset by the
effects of biochar on other soil processes, in particular prevention of N2O and CH4 release
from soil. Natural emissions of N2O from soil are a function of soil moisture status and
possibly tillage (Pekrun et al., 2003). Because biochar in soil may modify the moisture regime
and physical location of water within the soil matrix, it may mitigate the enhanced emission of
N2O that may occur in no-till systems. Methane emissions produced from agricultural soils,
mainly under paddy rice agriculture, account for 12% of the global methane emission from all
sources. Some studies have suggested that addition of biochar may partially suppress
methane emissions. The evidence for these effects is examined in Section 3.1.6.2.
There may be additional, potentially important offsets of other indirect emissions. These
could include avoided emission of CO2 if the fertiliser required to produce a tonne of product
is decreased via a positive effect of biochar on crop use efficiency. This extends to avoided
emissions of N2O during the manufacture of the fertiliser as well as from the soil upon
application – it is estimated that a carbon-equivalent emission of 1.8 tC is associated with
production and use of 1000 kg of fertiliser nitrogen (Mortimer, 2003). Any effect of biochar on
increased crop yield could significantly ease pressure on natural lands if implemented on a
large scale. Since the conversion of forest or savannah to agriculture can result in an
emission as high as 100 tC ha-1 (from above- and below-ground carbon stocks) this could be
important (Searchinger et al., 2008).
It has been suggested from visual observation of the terra preta that biochar could also lead
to a net stabilisation of other organic matter (Sohi et al., 2006; Lehmann and Sohi, 2008). If
this is the case, this could be another factor benefiting the overall net carbon gain from
biochar-based soil management strategies. This is a particularly intriguing prospect since the
capacity for soils to store biochar, not relying on protective capacity of a limited clay surface
area, is not finite in the way that it appears to be for other soil organic matter. Thus in
addition to representing a carbon store of its own, it is possible that biochar can enhance the
intrinsic soil organic carbon storage capacity of soil by affecting the turnover of indigenous
carbon. However, apparently contradictory data have been published, which seems to
suggest an accelerated decomposition of leaf litter in soil amended with biochar (Wardle et

19
al., 2008). The mechanisms and predictive description are, however, still to be determined
and defined.
A carbon balance for a pyrolysis scenario that appears positive with respect to the
atmosphere in the short term (i.e. a net emission) may be rendered negative if the feedstock
for biochar production comprises a new, additional and sustainably supplied resource. This
resource could derive from a managed increase in the above-ground productivity of existing
crops, or by maintaining vegetation in a state of high net primary productivity through growth
continuous cropping. Maintaining high net primary productivity above ground also promotes
higher productivity below ground, with an associated increase in root exudation and root
turnover. Plant and root productivity can also be enhanced by the use of fertiliser; noting,
however, the overall carbon balance of a strategy based on increasing soil carbon through
increased use of fertiliser. In assessing the carbon balance of a biochar strategy, it is
important to include the carbon cost of transporting feedstock and biochar between field and
pyrolysis facility and vice versa. In the scenario where there is no energy capture from
pyrolysis of organic matter, gains will depend on positive impacts on crop yield and soil
health.

3.1.3. Combined bioenergy and biochar production

Greatest overall impact on greenhouse gas emissions are likely to be realised where gases
produced during pyrolysis are captured and utilised in a manner that offsets fossil fuel
energy. In comparing a strategy that involves biochar production and its use in soil, it can be
argued that the reference scenario should be the use of the same feedstock in some other
form of bioenergy capture, such as simple combustion or, alternatively, pyrolysis with
gasification or combustion of the biochar by-product.
Bioenergy in general is often described as carbon neutral, since the carbon emitted in the
use of the energy approximates to the amount removed from the atmosphere to create the
feedstock in photosynthesis. For a biochar-based strategy to be carbon-negative, the
avoided carbon emission – or rather CO2 -equivalent emissions (since other greenhouse
gases have to be considered) – from use of 1t carbon in feedstock must exceed 1tC
(Renner, 2007). If it is assumed that the feedstock would otherwise decompose and return to
the atmosphere as CO2, as is the case for organic material added to soil, the carbon
emission from producing biochar and adding the stabilised residue to soil may alone, over a
few years, be close to one unit. Thus, with any net energy capture through use of gases (or
oils) produced during pyrolysis, the technology may be considered carbon negative
(Lehmann, 2007). The indirect effects on carbon emissions resulting from a positive impact
on agricultural productivity, plus an effect of biochar or the emission of non-CO2 greenhouse
gases from soil, may enhance this.
However, the carbon-negative status of the technology has been questioned by Bruun
(2008), who considered the additional fossil fuel offset that could be obtained if biochar was
used for non-energy purposes. Furthermore, biomass pyrolysis itself did not extract more
energy from a feedstock when compared to another bioenergy use. Bruun (2008) also
pointed out that over longer timescales, an increasing proportion of the carbon initially
sequestered in biochar would be slowly returned to the atmosphere through its slow
degradation back to CO2.
In terms of energy captured per unit of CO2 released it may be correct that biochar
production is not associated with less carbon emission than other forms of bioenergy.
However, when expressed in terms of energy captured per unit of carbon in the feedstock,
this is not the case. During pyrolysis the majority of energy embodied in feedstock (about
70%) is converted into combustible syngas, but with the liberation of only half of the
feedstock carbon (Lehmann et al., 2005a). This is because energy rich but less
carbonaceous functional groups are liberated first.
In the pyrolysis of organic wastes and crop residues, the emission avoided in preventing its
natural decomposition in soil, composting or landfill (for example) is an important part of the
overall CO2 -equivalent savings. In the utilisation of non-waste feedstocks, such as biomass

20
crops, there is no such avoided emission. Indirect CO2-equivalent savings resulting from use
of biochar in soil must then exceed the energy embodied in the biochar and allowance made
for the implications of land use.

3.1.4. Evaluation of biochar systems

Since pyrolysis is a more carbon-efficient way to capture bioenergy compared with other
bioenergy systems (in terms of CO2 MJ-1), manufacture and storage of biochar would add
significant benefits for climate change mitigation alone. From this perspective, storage of
biochar does not need to be in the soil, and it had been proposed that entire valleys could be
used as storage facilities for biochar (Seifritz, 1993). However, applying biochar to
agricultural soils is currently the most widely proposed path, since it is more likely to
overcome the opportunity cost in energy production (the recoverable energy forgone in the
biochar). If biochar can provide reliable agronomic benefit it may command a value in crop
production in addition to a potential carbon credit.
However, whilst the potential for management of the terrestrial carbon cycle is the reason for
the current interest in biochar, to be workable a biochar-based scenario must: (1) assess the
monetary value of direct and indirect emission savings arising from the use of biochar
against the opportunity cost of biochar combustion or alternative use, (2) provide certainty,
verification and possibly evidence for carbon-equivalent savings and (3) consider the indirect
costs and benefits to land users and upstream food processors from the use of biochar in
soil. The latter might include the cost of biochar application, weighed against the marketing
benefits gained through carbon-neutral food products.
In short, a full life-cycle analysis of alternative scenarios is required. However, greater
certainty is required on the following in order to fully assess biochar-based soil management
for specific applications: (a) the stability of biochar carbon in soil, (b) the indirect impacts of
biochar on carbon-equivalent emissions and (c) the security, reliability and constancy of price
for pyrolysis feedstocks. These are reviewed in more detail in the next sections.
The potential for technological developments in pyrolysis to enhance flexibility and overall
efficiency is a separate topic, and will be facilitated by its expansion and forums such as IBI,
and national networks such as the Network of Australian and New Zealand Biochar
Researchers, and the UK Biochar Research Centre. It should be highlighted, however, that
from the perspective of the economics of energy capture, the value of biochar and the overall
outcome of the analysis is sensitive to the price of heat and power generated from other
fuels. It is also affected by any subsidy for renewable energy, which may have the effect of
inflating the monetary value of the energy in biochar (Woolf, 2008).

3.1.5. Stability of biochar in soil

Its extraordinary stability means that charcoal particles in soil have been used as a tool for
dating and paleo-environmental reconstruction as well as evaluation of cropping practices
over centennial and millennial timescales (Ferrio et al., 2006, Scott et al 2000). Studies of the
age of the carbon in terra preta of Brazil, as well as similar black carbon accumulations in the
soils of other natural ecosystems that have resulted from natural fire events, provide
considerable reassurance for the general long-term stability of at least some significant
component of biochar. However, laboratory-based studies using freshly-made char tend to
show some mass loss – sometimes large – in a period of days to years.
The paradox of apparent long-term stability against measurable short-term decomposition
suggests that biochar comprises both stable and degradable components. At the moment
there is insufficient data in the literature to compare the responses between short- and long-
term stability under different climates and in different soils, which could enable the relative
size of these fractions to be assessed.
Combustion conditions during pyrolysis as well as the type of feedstock are probably
influential in determining the proportion of relatively labile components in biochar products.
Measuring the influence is essential for the optimisation of pyrolysis for maximum net carbon

21
stability. In optimising biochar production against energy capture to address climate change
most effectively, a consistent level of stability in biochar is the aim. Maximising retention of
carbon into biochar is counter-productive however, in both carbon and economic terms, if the
additional material is in fact associated with short- or medium-term loss to the atmosphere.
The chemical composition of biochar provides the principal explanation for its generally high
level of stability and is reflected in broad terms by its elemental composition: highly aromatic
and with a very high carbon content. It is likely that its stability is strongly modified by its
physical properties and structure, however. If the biotic and abiotic processes determining
the fate of biochar are the same as those for other soil organic matter, higher soil
temperature, moisture availability, lower clay content and intensive tillage will accelerate
decomposition rate.
The soil system has a remarkable propensity to degrade organic substrates introduced into
it. As a substrate generally very low in the concentration of key crop nutrients, the rate at
which biochar degrades in situ may also be influenced by the exudation of labile, nutrient rich
substances in the rhizosphere. This, in turn, is affected by cropping pattern.

3.1.5.1. Stability of biochar in natural systems

Soils that contain large amounts of char are those that have experienced relatively frequent
natural fires over a period of millennia (Lehmann et al., 2008). Converting a fraction of
standing plant biomass to black carbon in soil constitutes net removal of CO2 from the
atmosphere (Forbes et al., 2006). A relatively minor under-estimation in our estimates for the
percentage converted could explain up to one-fifth of the so-called missing carbon sink, that
is the imbalance between carbon eliminated from forest and fossil fuels, against observed
atmospheric CO2 (Kuhlbusch, 1998).
In natural systems it is not possible to determine the exact amount of biochar added to a soil
over the long-term history and the biochar in these soils will be of disparate age. However, a
loss rate constant can be derived mathematically by assuming that current and historic levels
of standing biomass are representative (Graetz et al., 2003; Mouillot et al., 2005; Forbes et
al., 2006), and a simple factor applied to capture the rate of conversion of biomass to
charcoal in burning episodes. This approach has yielded the best estimates for long-term
mean residence times, which are in excess of 1000 years (Lehmann et al., 2009). Direct
monitoring of archived soils from medium-term experiments seems to support such stability
(Skjemstad et al., 2001).
A portion of black carbon in soils globally also comprises condensed aromatic carbon in the
form of soot particles. This, in turn, can be confused with soot similarly formed in fossil fuel
combustion. Such black carbon is considered to lie at the most stable end of a black carbon
continuum (Masiello, 2004). Over extended timescales the physical transport of this material
through the soil and into water and sediment is inevitable and is seen in its accumulation in
marine sediment (Masiello et al., 1998). This observation highlights the potential to confuse
physical transport of biochar from a trial site for oxidative loss. It also indicates the
importance of such processes to the sequestration of carbon in the natural carbon cycle
(Smittenberg et al., 2006).
Although char from wildfire offers opportunities to study the long-term dynamics of pyrolysed
biomass, the fire and low rate of biomass conversion to char suggests probable differences
in composition and function. Simple charcoal manufacture probably began with the discovery
of fire; therefore, it is not surprising that sites of ancient habitation are associated with soils
enriched in char. Whilst the terra preta provide a neat and convincing example for the
deliberate use of biochar in agriculture, there is circumstantial evidence for its informal use in
other regions, not only in the distant past but in recent centuries (Young, 1804) and in the
current day (Lehmann and Joseph, 2009a).

22
3.1.5.2. Influence of biochar feedstock on stability

In soils where charcoal has been known to be abundant, it is associated mainly with the very
fine, sub-50 µm soil fraction (Brodowski et al., 2006). This suggests that through some
abrasive physical process or the destructive physical impacts of freeze-thaw and shrink-
swell, the initial size of brittle biochar particles may be relatively unimportant to its long term
interactions in soil. Ponomarenko (2001) found biochar evenly distributed across particle size
and biochar particles to become increasingly rounded over time at a rate dependent upon the
abrasiveness of the soil. Ponomarenko (2001) also found the concentration of clay-sized
mineral particles embedded in biochar pores to increase with time and noted that this would
inhibit their isolation using density-based techniques.
Brodowski (2006) found a small proportion of biochar particles in soil to be associated with
micro-aggregates, but for this biochar to account for a rather large proportion of the carbon
that is present in such structures i.e. compared to other particulate organic matter. The
authors used this as evidence for physical protection of biochar against degradation and also
noted that biochar might act as a binding agent for organic matter in aggregate formation.
Although they did not specify whether the latter would be a purely physical interaction or a
consequence of biological activity, Watts (2005) previously found no effect of charcoal on
aggregation at low temperatures designed to preclude biological activity. In an earlier
publication Glaser (2000) found a large proportion of biochar in terra preta to be present in
unprotected fractions. Obtaining similar results, Murage (2007) noted that the misleading
impression might be gained that active soil fractions turn over more slowly in soils that are
enriched in biochar.
Currently there is no published information to demonstrate whether the physical diminution of
biochar in soil is accompanied by oxidative loss of carbon. However, X-ray photoelectron
spectroscopy showed abiotic oxidation (with proliferation of carboxyl groups) to occur in the
porous interior of biochar, whilst biotic oxidation affected external surfaces only (Cheng et al.,
2006). Biotic oxidation might therefore be enhanced as particle size decreases, although in
the study of Cheng (2006) it was also quantitatively less important. These findings have been
further validated (Lehmann et al., 2005b) using synchrotron-based techniques (near-edge X-
ray absorption fine structure spectroscopy).
At the macro-scale biochar products range from powdery to brittle, depending upon the
physical microstructure of the material from which they are derived. Those produced from
woody feedstock display a predominantly xylemic structure that is coarse and strong. These
also display the highest carbon contents, in excess of 70% C and up to 90% C, and are low
in trace elements. Those produced from rye grass, maize and digested feedstocks i.e.
manures are powdery, lower in C (over 60%), and enriched in minerals and nutrients. Thus
the latter are not only less physically recalcitrant, they are also a more attractive microbial
substrate.

3.1.5.3. Climatic effects on biochar mineralisation

Climate determines soil temperature, which affects the rate at which both biotic process and
abiotic reactions occur in the soil. However, in the absence of water the effect of temperature
is irrelevant, as water is essential for biological cell function, and solution phase reactions
proceed by definition only where water is present. Thus synchrony in a conducive
environment of rainfall and temperature and rainfall conditions is required in order to
maximise overall soil activity.
Biochar should modify not only the soil water holding capacity of the bulk soil, but also the
physical location of water within the soil matrix, since the smallest pores become water-filled
first and remain moist the longest (Gaskin et al 2007). The size of these pores makes them
relevant to microbial populations as a physical niche. There is also evidence that mean soil
temperature and diurnal temperature fluctuations are impacted by the effect of biochar on
soil colour (Krull et al., 2004; Oguntunde et al., 2008).

23
The effect of climate on biochar oxidation has been evaluated in a transect study spanning
several climatic zones. Sites along the transect had each received inputs of biochar during a
brief and defined period in industrial history 130 ybp, but received greatly contrasting mean
annual temperatures (Cheng et al., 2008). The accumulation of cation exchange capacity
was correlated to mean temperature and the extent of oxidation was seven times higher on
the external surfaces of biochar particles than in the interior.
The potential consequence of a warmer global climate releasing CO2 through accelerated
mineralisation of black carbon was first scoped by Druffel (2004) who indicated potential
emission rates of 2 to 7 GtC y-1 by 2100. However, if a larger proportion of the global soil
carbon stock comprises recalcitrant black carbon than is assumed in current models, the
more active pools must be smaller and the overall response to warming significantly slowed
(Lehmann et al., 2008). General arguments concerning the relative response of recalcitrant
carbon and more active carbon pools to changing temperature has not been resolved (Fang
et al., 2006).

3.1.5.4. Soil biological activity and the stability of biochar

A number of relatively short-term laboratory incubation experiments have been conducted

using soil mixed with biochar, with or without added substrate, to assess the biological and
chemical degradation of biochar to CO2. Early studies found that even graphitic carbon could
significantly be mineralised biologically (Schneur, 1966).
Laboratory studies where loss routes can be controlled have generally found some
measurable initial loss of carbon from biochar (Cheng et al., 2006); however, a few studies
have found a much larger loss (Hamer et al., 2004). In the Hamer (2004) study, not only was
charcoal substantially degraded within 60 days, but the mineralisation of simultaneously
added glucose solution was also enhanced, suggesting a priming effect. Combined products
comprising labile nitrogen and stable carbon have been proposed (Day et al., 2005).
However, the evidence (Hamer et al., 2004) suggests that the addition of nitrogen could
impact on the stability of biochar and its value in carbon sequestration. This has not,
however, been experimentally assessed to date.
Further studies seem to indicate that biochar may prime the decomposition of native soil
organic matter. After nine years in the field, Wardle et al. (2008) measured mass loss greater
in a charcoal-litter mixture, than for the sum loss in a charcoal-only plus litter-only. However,
the mesh bags in which the samples were contained had been buried in a forest litter layer,
and thus separated from mineral soil. Although the loss could have been from the priming of
litter decomposition, it was also possible that the loss could result from priming of charcoal
loss (Lehmann and Sohi, 2008).
If the level of organic matter input to the soil by plants is steady for a particular ecosystem,
enhanced microbial activity over the long term is not consistent with a net accumulation of
non-black organic matter (as well as charcoal) that is observed in terra preta, compared to
their adjacent soils. But in fact these observations need not be contradictory if carbon that is
mobilised in the litter layer is, once transported to the mineral soil, rapidly stabilised. The
latter effect could directly involve biochar surfaces or minerals in subsoil, as well as mineral
material in the surface horizon (Lehmann and Sohi, 2008).

3.1.5.5. Effects of biochar on ease of tillage and mechanical disturbance

If, as proposed, natural soil movement influences the breakdown of biochar through its
reduction in size, then the rate of breakdown would be expected to be further accelerated by
tillage. This is important to consider since tillage is perhaps envisaged as the primary means
to incorporate biochar into soil.
Quénéa (2006) reported a 60% decrease in both the soot and charcoal content of sandy soil
under temperate forest during 22 years after conversion to intensive agriculture with annual
tillage. The loss of total soil carbon over the same period was 30%, suggesting that biochar
and charcoal were relatively less resistant to degradation than bulk soil organic matter after

24
disturbance. However, the analysis of the charcoal data was based on larger hand-picked
fragments and it seems likely that particles broken down into very fine fractions might have
led to an overestimate of the loss. The initial soot content was very low. In contrast, 50 years
of cropping and cultivation had no measurable change in the aromatic aryl carbon, taken to
reflect charcoal, whilst other fractions declined rather rapidly (Skjemstad et al., 2001).
More research may enable likely rates of breakdown to be predicted. If, for example, tillage
was a key factor, maximum longevity of biochar targeted by application on land where
minimum tillage is practised. In no-till systems, biochar could be sequestered into soil
through a one-time addition at the time of conversion from a tilled system.

3.1.6. Indirect impacts of biochar on CO2-equivalent emissions

The net carbon gain resulting from stabilisation of carbon into biochar, and its storage in soil,
needs to be refined. There is even less information available for the impact of biochar in soil
on the emission of greenhouse gases other than CO2. The contribution of nitrous oxide (N2O)
and methane (CH4) are major contributors to climate forcing and have significant agricultural
sources, including soils. There is evidence that biochar may suppress the emission of these
gases from soil.
There may be additional indirect benefits in terms of greenhouse gas emissions, resulting
from improved crop production through use of biochar. Although the global possibilities are
currently hypothetical, in an era of falling global grain stocks, the value of biochar could
extend beyond a purely financial consideration for land users and become increasingly
relevant politically and economically. Growing concerns around both energy and food
security may accelerate the development and application of biochar technology and
associated Governmental or inter-Governmental market interventions to support it.

3.1.6.1. Indirect effects of yield benefits and nutrient requirement

The direct impacts of biochar on crop yield are reviewed in Section 3.2.2.
Where economically optimal fertiliser rates are currently applied, biochar has the potential to
deliver the same crop yield with a lower application rate – with potentially significant
greenhouse benefits. In more detailed assessments for the overall carbon balance of a
biochar strategy (Gaunt et al., 2008), an assumed 10% reduction in the fertiliser required to
maintain current crop yield was found to be a particularly important component of the net
carbon benefit. This reflected the energy intensive nature of nitrogen fertiliser production and
the N2O emissions that result from fertiliser application and use.
The purpose of biochar application might not simply be to attain a greater yield, however, but
possibly to achieve predictability in yield through a lower susceptibility to climatic events such
as floods and drought.
Also, the economic optimum after biochar application could be gained through a gain in crop
yield at the same current or possibly higher rate of application, in which case the net result
would be higher per hectare yields.
Furthermore, biochar could increase, maintain or at least limit gradual decreases in crop
yield on land where soil fertility and productivity is currently in decline. Under changing
climate the benefit of biochar in response to increasingly erratic or intense rainfall events
could be more acute, and/or enable plants to better exploit higher CO2 concentrations. This
suggests a potential role for biochar to assist in adaptation to climate and environmental
change.
These factors would benefit global carbon balance in several very important ways. Firstly it
could reduce the degradation of existing agricultural land, and thus alleviate pressure on
natural systems, which usually represent a significant store of carbon as well as biodiversity.
Maintaining or enhancing productivity of existing land may also make relatively more land
available for bioenergy or other alternative crop production systems.

25
Modelling of these possibilities, at this stage, may not be possible due to too many
uncertainties (economic as well scientific). However, if we view biochar as a technology with
the potential to be practised on a very large scale, they have to be assessed in the long term.
Importantly, they highlight the complexity of the biochar topic and the need to think carefully
about the system boundaries drawn in assessing benefits and impacts.

3.1.6.2. Impact of biochar on nitrous oxide emission from soil

The global warming potential (GWP) of a gas reflects two aspects: the efficiency of the
molecule in absorbing incoming solar radiation and its rate of chemical breakdown in the
atmosphere. By definition the global warming potential (GWP) of CO2 is 1.0; by comparison
the GWP of nitrous oxide is 310. Under anaerobic conditions N2O is emitted from soil through
denitrification, a process in which specialised microbes that obtain energy from the reduction
of nitrate (NO3-), or intermediate gases, to dinitrogen (N2). However, it appears that nitrifying
bacteria generally involved in conversion of N2 to ammonium (NH4+), i.e. nitrification, may
simultaneously denitrify (Bateman, 2005).
The availability of NH4+ is generally controlled by organic matter mineralisation, which is
climate driven, but its concentration is greatly enhanced by the application of nitrogen
fertiliser or, in livestock and gazed grassland systems, from dung or slurry application.
Regardless of the setting or source, the majority of soil nitrogen is in organic form and N2O
emanates from the utilisation of a relatively small and dynamic nitrogen pool.
Life cycle assessments quantifying the benefits of biochar-based strategies for energy
depend quite heavily on a decrease in the emission of N2O that frequently follows the
addition of mineral nitrogen fertiliser. Accounting for this effect makes a great difference to
the overall analysis of how a biochar to soil strategy impacts on net greenhouse gas balance
(Gaunt et al., 2008). The expectation for this effect relates to the general impact of biochar
on retention of N in the soil in a way that also enhances crop nutrition (see Section 3.1.6.1.).
If the latter effect is surface-mediated, it seems unlikely that biochar confines N to a physical
location – such as very small pores – where it is inaccessible to denitrifying bacteria. It may
be that, instead, biochar inhibits the process by sequestering dissolved mineral N.
Published data demonstrating the effect of biochar on suppression of N2O remains very
limited. In the most widely cited study to date (Yanai et al., 2007) ‘bio-waste’ charcoal was
applied during a re-wetting of a former grassland soil, high in organic matter, in laboratory
incubation (25°). Nine-tenths of N2O was suppressed in five-day emission episodes after
wetting of soils to 73% and 78% water filled pore space. At a slightly higher water filled pore
space (83%), charcoal had the opposite effect, increasing N2O emission. The rate of biochar
addition used in the study equated to a relatively high application rate of 180 t ha-1 in topsoil.
However, the authors were able to exclude the possibility that the alkalinity of their charcoal,
or its nutrient content, were significant factors in their observations.
In an arable soil with much lower C content (2.2 %C), Sohi (2008) has studied the effect of
willow charcoal at a much lower rate of 10 t C ha-1 which were assessed during 20°C
incubation of wet (70% water holding capacity) and re-wetted (from 20% water holding
capacity) soils, with and without simultaneous addition of small amounts of inorganic N
(equivalent 75 kg N ha-1). A more modest suppression of 15% was proportionally similar for
all treatments where there was any response at all (the already-wet soil did not emit
significant N2O). After six months, available soil N would have been largely consumed and
the soils thoroughly equilibrated. A second inorganic N addition (without new charcoal) at this
time showed no difference in N2O emissions between amended and control soils.
If any correspondence exists between the two studies it appears that not only is effect of
biochar on N2O likely to be non-linear with respect to rate of application (and significant but
not large at realistic rates) but – as authors of both papers conclude – the effects are likely to
reflect the impacts of biochar on soil physical properties, particularly modification of pore-size
distribution (of which water holding capacity is not a sensitive measure). In particular the
effect may only be seen during re-wetting, and not when soils are maintained wet. Whether
the benefit upon re-wetting is repeatable remains to be established.

26
Measurements of N2O emission in the field environment are difficult due to the transient and
spatially variable nature of denitrification. Like the studies described in Chapter 3, the
availability of sample biochar in the quantities required to assess its many effects in true
randomised plot designs presents a major challenge. Biochar field experiments with periodic
measurement of N2O in cover boxes is currently in a third year in upstate New York
(<http://www.css.cornell.edu/faculty/lehmann/research/biochar/biocharproject.html>).
In tropical environments field experiments have been established in Columbia and in Kenya.
Results from the Columbian trials indicate 80% suppression of N2O emissions (Renner,
2007).

3.1.6.3. Impact of biochar on methane emission from soil

Methane has a relatively low GWP of 21, but is six times more abundant in the atmosphere
than N2O (1.8 ppm compared to 0.3 ppm for methane), and has an annual flux approximately
50 times higher. Aside from industrial emissions, including natural gas exploitation and
distribution (accounting for about 20%), methane emanates primarily from the soil of natural
habitats and thus, uniquely for the main greenhouse gases, increasing rates of emission
have begun to stabilise. Within agriculture, paddy cultivation of rice and the guts of the
growing ruminant population (primarily grazing domestic animals) are key methane sources
(IPCC-2001,
<http://www.grida.no/publications/other/ipcc%5Ftar/?src=/climate/ipcc_tar/wg1/index.htm>)
Specialised methanotrophic bacteria make most aerobic soils a net sink for methane,
however, and the relatively rapid consumption of methane explains to a large extent its
relatively low GWP. The link between methane consumption and aerobicity is important in
the context of reduced tillage, however. These practices have been widely promoted for their
sequestration of carbon. Whilst higher topsoil organic matter is liable to improve both water
infiltration and holding capacity, increased moisture status is likely to result in a relative
increase of methane emissions (e.g. Castro et al., 1995).
Field experimentation with biochar in Columbia showed the elimination of CH4 emission
(Renner, 2007). A number of studies are currently assessing the impact of biochar on the
emission of methane from paddy soils. At this time there are no results published in the
scientific literature.

3.1.6.4. Biological activity and stabilisation of soil organic matter

The stability of biochar and the biological activity that results from its application are
intrinsically linked, as are soil properties such as clay content, pH and cation exchange
capacity (CEC) and climatic variables.
The ancient terra preta are higher in organic matter compared with adjacent soils that do not
contain black carbon (Lehmann et al., 2003). This has lead to the hypothesis that black
carbon in soil leads to increased stabilisation and hence accumulation of other carbon. This
may provide one of the few mechanisms by which the intrinsic capacity of a soil to store
organic matter can be modified from a management perspective, and if correct, increases the
net carbon gain from the use of biochar.
However, some studies have reported increased microbial activity in soils enriched in biochar
(Steiner et al., 2003; Steiner et al., 2008). Upon addition of biochar to soil for the first time,
mineralisation may be stimulated by the presence of an active fraction and associated
soluble nutrients or labile carbon fractions. It has also been noted that the physical structure
of typical biochar products provides a secure environment for microbial colonies (Ogawa,
1994). However, it should be noted that experiments must be designed such that the system
is correctly monitored, e.g. all soil layers, gaseous losses and plant growth. Currently,
findings to date should be viewed as provisional.
Microbial biomass is not a measure of microbial activity but the abundance of microbial cells.
Thus whilst an increased microbial population associated with increased soil organic matter

27
without any obvious increase in substrate supply may seem paradoxical, it might suggest a
decrease in microbial efficiency, possibly associated with a change in the balance between
different groups of microbes. A simultaneous increase in CO2 emission, however, would
indicate enhanced plant growth and higher inputs of labile carbon via the plant roots.
In soils where char is present naturally in higher concentrations as a result of natural forest
fire or prescribed burning, the microbial community may include specialised species with a
capacity to degrade a relatively scarce and challenging substrate. This may explain the
distinct communities observed in terra preta, but is not clear whether these species are truly
ubiquitous to other soils (if scarce), or whether the capacity for soils to degrade such
materials is acquired over relatively long periods.

3.1.6.5. Effects of biochar on tillage and irrigation requirements

Lower bulk density and/or higher organic matter reduce the fuel requirement for mechanical
tillage, which can be managed in few other ways. It may also facilitate the process of
reducing the tillage used in agricultural systems. Soils with higher organic matter content also
tend to display higher water-holding capacity. Thus a further saving in energy costs can
come from reduced irrigation frequency or intensity. Surprisingly little information has been
collected on the impact of biochar on such parameters. However, in Ghana, kiln sites
showed topsoil bulk density approximately 10% lower than in adjacent soils (Oguntunde,
2008).

3.1.7. Biochar scenarios for agriculture

In addition to the price of biochar, land-users will incur the direct costs of applying biochar to
the soil. There are also potential non-monetary costs associated with the collection of straw
from their land as a pyrolysis feedstock, which affects the readiness of land owners to
engage in the market. In ‘closed’ systems where biochar is returned to the same land that the
feedstock originated, there may be opportunity costs. Currently, no socio-economic studies
exist that would address questions on these matters. However, one key advantage of a
biochar strategy is that, assuming that the provision of key functions is limited only by the
longevity of the biochar, its stability would dictate that annual or even regular applications
would be unnecessary to obtain benefits.
Widespread use of non-waste feedstocks for energy and biochar (or only biochar) could
impact not only commodity prices but, in a manner analogous to that seen with large-scale
bio-ethanol production in the USA, impact on the economics of continued energy production
through feedbacks on land and input prices. This raises complex socio-economic issues that
must be considered (The Royal Society, 2008) and modelled (Rokityanskiy et al., 2007).
Also, as for any assessment of bioenergy systems, it is essential to define boundaries that
spatially allow all possible land-use effects to be assessed, in the context of the overall net
greenhouse gas benefit (Searchinger et al., 2008).
The proximity of a pyrolysis facility to an adequate catchment for feedstock must be
economically and logistically viable, and can potentially affect the CO2 -equivalent savings.
This is the case for biomass and bioenergy facilities generally. However, for biochar the
proximity of suitable locations for biochar application to soil is important as well. If the
gathering of feedstock and distribution of biochar occur over the same area, the logistical and
cost impacts may not be greatly affected. However, it is important to think about biochar
scenarios in a spatial context (Figure 10).

28
Figure 10. Scenario for multi-feedstock production of biochar, and multi-application use,
emphasising the spatial context

Possible off-farm resources include municipal green waste from gardens and parks,
composted or compostable urban waste, digested sewage sludge and mixed municipal
waste. In addition, in the future, by-products of other bioenergy or bio-fuel systems may be
available.
Utilisation of off-farm wastes for biochar production holds the attraction of potential cost
savings from avoiding landfill or other disposal charges. In addition, compared to typical or
existing disposal methods, there may be a lower emission of CH4 and N2O greenhouse
gases than that emanating from direct placement in soil, enhancing the net gain in carbon
equivalents through avoided emissions of high GWP gases. However, many such wastes
have a high water content which will incur increased emissions (and cost) associated with
higher requirement for process energy in pyrolysis.
In a ‘closed loop’ scenario, biochar is incorporated into the same land, or at least the same
enterprise or groups of enterprises, from which the pyrolysis feedstock originates. A typical
scenario would involve utilisation of cereal crop straw that in intensive arable areas is often,
effectively, a waste product. Although there is no published laboratory work to support the
use of biochar produced from wheat straw, there is limited existing information on the relative
stability of biochar from rice husk, sugarcane bagasse and straw from maize. In industrial
agriculture crop straw may constitute 2 t C ha-1. Putting this scenario in the context of the UK

29
example, with approximately 4 M ha of cereal crops grown, annual total fossil fuel CO2
emissions amount to 170 M t.
Theoretical comparisons have been made for the carbon-equivalent gain offered by the
pyrolysis of maize straw versus use of a dedicated biomass crop (Gaunt et al., 2008).
Utilising biomass crops for energy on a large scale has a potential impact on the land
available for food production and may exert new pressure on non-agricultural land use.
Biochar produced from the pyrolysis of biomass crops might be incorporated into different
agricultural land from where the biomass feedstock was grown. This could be for greater
agronomic gain, to apply the product in rotation over a wider area to maximise benefits, or to
deal with the cumulative quantity of product. In the combined energy and bio-oil co-
production case study considered by Ogawa (2006), the biochar by-product was also
returned to adjacent arable land.
Most scenarios considered to date have focused on conventionally managed arable land,
where biochar could be added to soil as part of an existing tillage regime. Biochar could be
incorporated during conversion of land to no-till, but a strategy of integration into no-till and
grazed grassland systems has not yet been considered.

3.1.7.4 Forestry

Ogawa (2006) outlined a scheme for carbon sequestration by forestation and carbonisation.
This scheme revolved around fast growing plantation tree species fixing atmospheric CO2,
with the products comprising not only conventional timber, wood chip, and pulp, but diversion
of wastes and residues to a carbonisation procedure and re-application of this stabilised
carbon back to the plantation soil. This approach has been formally proposed under the
Clean Development Mechanism for a scheme in Sumatra, Indonesia.
In the state of Minas Gervais, Brazil, an existing commercial project claims, under CDM, a
carbon credit for substitution of coal-derived coke for smelting of iron by pyrolysed plantation
eucalypt. The project produces 300,000 t y-1 charcoal. The charcoal ‘fines’ that account for
about 5% of the product are utilised for briquette manufacture rather than application into
soils. In Australia, the potential for integration of oil production from oil mallee trees with
processing of wood waste for production of biochar for use in crop production has been
examined (McHenry, 2009).
Seifritz (1993) evaluated the size and cost of the carbon gain that could be realised by
straight conversion of plantation forest biomass to stockpiled biochar. The scenario included
no capture of energy in the conversion, highlighting instead the net primary productivity that
is maintained by cropping, and the long lifetime of charcoal compared to the nature and fate
of traditional timber products.
In the tropical context, ‘slash and char’ scenarios have been discussed, where one-off inputs
of biochar are made during conversion of land from forest to agriculture (Steiner, 2006), or
perhaps ‘crop and char’, with cycle of positive feedback between one-off, occasional or
rotational inputs of biochar and increasing biomass productivity and feedstock resource. In
both cases, pyrolysis would be performed using the most basic (and perhaps relatively
inefficient) technology such as simple pits of clay kilns. However, as examples of viable,
village-scale bioenergy based on gasification technology in developing countries increase, it
is conceivable that technological development in tandem with increased income from ‘crop
and char’ practices might ultimately realise combined biochar production and energy capture
at the same scale.
In the absence of adequate technological development, charring and manufacture of
charcoal may not offer the same benefits to human health as, for example, the substitution of
existing biomass burning practices for basic but cleaner and more efficient combustion
technology (Wang et al., 1999). Conventional charcoal production may also release methane
and other trace greenhouse gases (Edwards et al., 2003). Sub-micron soot particles
produced by condensation reactions in gas streams from combustion comprise the most
recalcitrant forms (Figure 7) of black carbon but, despite the relatively small quantities of

30
carbon involved, may be having an important impact on the albedo of both the global
atmosphere and ice caps, altering the radiative balance and exacerbating climate change
(Ramanathan et al., 2008). Currently the global emission of soot is predicted to decline as
rural users of biomass in developing countries switch to clean burning fossil sources (Streets
et al., 2004). Charcoal manufacture produces less soot than open burning, but despite the
possible scale of future biochar production, its future contributions to the global soot
inventory has not been formally examined.

3.1.8. Notes on the natural cycling of char in soil

Observation shows that wildfires are a routine natural and indeed often essential feature of
many natural grassland, forest and woodland ecosystems, and fire is gradually being
incorporated into global ecosystem models (Thonicke et al., 2001). Fire frequency does not
appear to decrease soil C by affecting plant productivity (Ansley et al., 2006) and may
increase it (Czimczik et al., 2005; Ansley et al., 2006); fire does not destabilize black carbon
already present (Ansley et al., 2006) or only partially in organic boreal soils (Czimczik et al.,
2005). Studies using remote sensing have attempted to quantify the pattern and frequency of
burnings (Seiler et al., 1980) and most estimates suggest approximately 1-5% of standing
biomass is converted to black C (Schmidt et al., 2000; Forbes et al., 2006). This figure is
much lower than the figure previously proposed by Kuhlbusch (1996), who suggested
sequestration into black C during the relatively recent era of forest clearance could explain
up to 20% of the so-called missing carbon sink. A detailed analysis in Australia has
suggested that natural fire might provide a sink equivalent to 8.3 MtC yr-1 (Graetz et al.,
2003).
In fired-affected systems standing biomass remains in equilibrium, viewed over the long
term. However, a modest rate of stable charcoal formation during burning means that the net
result of natural fire is that the carbon content of the wider system, including the soil, should
gradually increase. However, this is a slow process and difficult to measure in the field with
repeated experimental burning (Dai et al., 2005). Given that the net primary productivity of
biomass (60 GtC yr-1) is quite large relative to the soil C pool (1500 Gt) the black C pool
should become dominant in the soil over geological time if this fraction did not degrade at all,
even with occasional fire and a low charcoal conversion rate (Graetz and Skjemstad, 2003).
Thus the fact that soil carbon dynamics can be modelled by postulating a relatively small
inert- or very slow-turnover pool (Falloon and Smith, 2000; Smith et al., 2000) is an indication
that biochar must degrade, at some slow rate, even once transport into water and estuarine
sediments is considered (Schmidt, 2004; Simpson and Hatcher, 2004b; Smittenberg et al.,
2006) and accounted for (Masiello et al., 1998).
The Roth-C soil carbon model explicitly incorporates char as one of its pools (Falloon and
Smith, 2000). The inert pool in the Roth-C soil carbon model represents more than simply
char, but also exceptionally degraded, highly stabilised organic matter (Falloon and Smith,
2000). Radiocarbon dating can be used to experimentally refine the site-specific size of the
inert pool to model total soil carbon. However, estimates for a range of sites where such data
are available has suggested only a general relationship with soil texture (Falloon et al.,
1998). It is likely, however, that long-term field experiments of duration useful in
parameterisation of soil C models will provide the data needed to improve on the char pool in
the model.
It may be significant that one experimental site where the model does not simulate field
measurements well is the site of the Waite plots in Australia, where there is a documented
history of burning (Coleman et al., 1997). Also in Australia, Skjemstad (2004b) demonstrated
that by re-allocating carbon between soil pools according to a direct measurement of char,
the Roth-C model may simulate the trajectory of carbon for a range of soils with similar
burning history.
In addition, new techniques for quantifying char experimentally (see Section 2.2.3.) reveal
that, when applied to a wider variety of sites, observed levels of char were considerably
higher than accommodated by the modelled inert pool (Schmidt et al., 1999; Preston et al.,

31
2006). The implications of this finding for our prediction of climate change feedbacks from
enhanced decomposition will become important, as soil models are increasingly linked into
global climate modelling (Lehmann and Joseph, 2009b).
Unfortunately the most reliable and direct techniques for quantifying char are currently not
sufficiently practical for application at the scale useful to assessing the carbon stock that it
represents, at the relevant (global) scales. However, the development of new spectral
analysis techniques using mid-infrared wavelengths (Section 2.2.3) may lead to an approach
that is both rapid and low cost (Janik et al., 2007).

3.2. Biochar, crop productivity and resource management

Conceptually three main mechanisms have been proposed (described in detail below) to
explain how biochar might benefit crop production: (i.) direct modification of soil chemistry
through its intrinsic elemental and compositional make up, (ii.) providing chemically active
surfaces that modify the dynamics of soil nutrients or otherwise catalyse useful soil reactions,
(iii.) modifying physical character of the soil in a way that benefits root growth and/or nutrient
and water retention and acquisition.
The first mechanism may result in a temporary change in crop productivity, the size and
duration of which will be dictated by the natural process of biochar weathering and the effects
of crop off-take. This could occur where the biochar has significant mineral nutrient content,
or conversely increase in CEC over time as the biochar weathers. If biochar releases these
elements, then establishing the fate of biochar carbon during this release is extremely
important in the context of the underlying rationale for biochar production and application in
soil.
Benefits provided through the second and third mechanisms depend on the long-term
physical persistence of biochar and may thus also be finite, although over a much longer
timeframe. This would include the impact of porous biochar on water retention or lowering
soil bulk density. The magnitude and the relative importance of the three mechanisms in a
particular setting will evolve over time as the slow process of chemical and physical
modification results in a gradually increasing concentration of smaller, partially-oxidised
particles. Evidence for the general resistance of biochar to chemical and biological oxidation
is addressed in an earlier section (3.1.5.).
Quantitative evidence for the stability of biochar does not equate to constancy in functional
characteristics, since the chemical properties of biochar itself have been shown to develop
over time, with implications for functional interactions in the soil environment. In a key multi-
parameter study Cheng (2008) showed that properties that become enhanced over time are
CEC and pH, as a result of gradual surface oxidation (Section 3.1.5.). The size of biochar
particles is relatively rapidly decreased, concentrating in size fractions <5µm diameter
(Sections 3.1.5.1. and 3.1.5.5.).
In assessing the agronomic performance of biochar, comparisons should be made against
the properties of both the same feedstock un-pyrolysed and alternative biochar produced
from other feedstocks. If the feedstock is produced from the same land to which the material
is returned (e.g. cereal straw), biochar would not normally substitute all deliberate returns of
organic matter to a soil, but rather a one-time or occasional amendment. This is important
since as previously noted (Section 3.1.1.) soil fertility depends on degradation of organic
matter, and the recycling of plant nutrients. Biochar should be viewed as a mechanism to
enhance that process through its moderation, and not its termination. In any case, the
material potentially used in pyrolysis, is roughly matched by the amount of labile organic
matter exuded into the soil by the plant roots.

3.2.1. Soil fertility

32
Expectation of increased soil fertility benefits arise from studies of the terra preta that
contains high proportions of black carbon (Haumaier et al., 1995; Glaser et al., 2002;
Lehmann et al., 2003; Lehmann and Rondon, 2006). The evident fertility of the terra preta is
generally attributed to high soil organic matter content – organic matter assists in the
retention of water, soil solution and cations – and the retentive capacity of aged biochar itself
for nutrients and water (Section 3.2.3 and 3.2.4),
The black carbon present in terra preta is thought to originate from partially-combusted
biomass residues derived from a range of anthropogenic activities, including kitchen fires and
field burning. A particularly striking characteristic is a stronger relationship between soil
carbon content and soil CEC in these soils relative to adjacent land, indicating that biochar
comprises a greater proportion of soil carbon (Liang et al., 2006). Since CEC is indicative of
the capacity to retain key nutrient cations in the soil in plant-available form and minimise
leaching losses (Section 3.2.4), this is cited as a key factor where differences in crop
productivity are observed.
High rates of biochar addition in the tropical environment have been associated with
increased plant uptake of P, K, Ca, Zn and Cu (Lehmann and Rondon., 2006). In contrast to
mainstream chemical fertilizer, biochar also contains bioavailable elements such as selenium
that have potential to assist in enhancing crop growth.
There has been much speculation concerning the potential effects of biochar on microbial
activity in soil, which in the context of terra preta has been reviewed in detail by Steiner
(Steiner et al., 2003). Assuming that plant inputs and hence microbial substrate remain
unchanged, enhanced microbial activity alone would diminish soil organic matter. However,
this is contrary to the observation in terra preta, where soil organic matter is generally higher
than in similar surrounding soil (Liang,2006). However, a change in the balance of microbial
activity between different functional groups could benefit crop nutrition, specifically
enhancement of mycorrhizal fungi (Ishii et al., 1994), and this could feed back into higher net
primary productivity and carbon input.
There is relatively extensive literature documenting stimulation of indigenous arbuscular
mycorrhizal fungi by biochar, and this has been reflected in plant growth e.g. Rondon (2007),
Nishio (1996). This literature has been reviewed in some detail by Warnock (2007), who
proposed four mechanistic explanations, of which a combined nutrient, water and CEC effect
was considered most probable.

3.2.2. Crop yield

The majority of currently published studies assessing the effect of biochar on crop yield are
generally small scale, almost all short term, and sometimes conducted in pots where
environmental fluctuation is removed. These limitations are compounded by a lack of
methodological consistency in nutrient management and pH control, biochar type and origin.
Studies in a wide range of climates, soils and crops have been conducted. It is not therefore
possible at this stage to draw any quantitative conclusion, certainly not to project or compare
the impact of a particular one-time addition of biochar on long-term crop yield. Nonetheless,
evidence suggests that at least for some crop and soil combinations, moderate additions of
biochar are usually beneficial, and in very few cases negative.
Glaser (2001) reviewed a number of early studies conducted during the 1980s and 1990s.
These tended to show marked impacts of low charcoal additions (0.5 t ha-1) on various plant
species. Higher rates seemed to inhibit plant growth. In later experiments, combination of
higher biochar application rates alongside NPK fertiliser increased crop yield on tropical
Amazonian soils (Steiner et al., 2007) and semi-arid soils in Australia (Ogawa, 2006). Due to
the year to year variation in climate and its impact on short-term dynamics, results from a
number of field experiments recently set up are, whilst generating data, not yet published.
The nature and mechanistic basis for interactions between crop, soil type, biochar feedstock,
production method and application rate will have to be understood to gain predictive capacity
for the performance of biochar in soil, and open the possibility for large scale deployment.

33
Table 5. Summary of experiments assessing the impact of biochar addition on crop yield
Authors Study outline Results summary
Iswaran et Pea, India 0.5 Mgha-1 char increased biomass
al (1980)* 160%
Iswaran et Mung bean, India 0.5 Mgha-1 char increased biomass
al (1980) * 122%
Kishimoto & Soybean on volcanic ash 0.5 Mgha-1 char increased yield
Sugiura loam, Japan 151%
(1985) * 5 Mgha-1 char decreased yield to
63%
15 Mgha-1 char decreased yield to
29%
Kishimoto & Sugi trees on clay loam, 0.5 Mgha-1 wood charcoal
Sugiura Japan increased biomass 249%
(1985) * 0.5 Mgha-1 bark charcoal increased
biomass 324%
0.5 Mgha-1 activated charcoal
increased biomass 244%
Chidumayo, Bauhinia trees on Charcoal increased biomass by
(1994}* alfisol/ultisol 13% and height by 24%
Glaser (2002) Cowpea on xanthic ferralsol 67 Mgha-1 char increased biomass
150%
135 Mgha-1 char increased biomass
200%
Lehmann (2003) Soil fertility and nutrient retention. Bio-char additions significantly
Cowpea was planted in pots and rice increased biomass production by 38
crops in lysimeters at the Embrapa to 45% (no yield reported)
Amazonia Ocidental, Manaus, Brazil
Oguntunde (2004) Comparison of maize yields Grain yield 91% higher and biomass
between disused charcoal yield 44% higher on
production sites and charcoal site than control.
adjacent fields.
Kotokosu watershed, Ghana
Yamato (2006) Maize, cowpea and peanut trial in area Acacia bark charcoal plus fertiliser
of low soil fertility increased maize and peanut yields
(but not cowpea)
Chan (2007) Pot trial on radish yield in heavy soil 100 t ha-1 increased yield x3; linear
using commercial greenwaste biochar increase 10 to 50 t ha-1
(three rates) with and without N - but no effect without added N
Rondon (2007) Enhanced biological N-2 fixation (BNF) Bean yield increased by 46% and
by common beans through bio- char biomass production by 39% over the
additions. Colombia control at 90 and 60 g kg(-1) bio-
char, respectively.
Steiner (2007) Four cropping cycles with rice (Oryza Charcoal amended with chicken
sativa L.) and sorghum (Sorghum manure amendments resulted in the
bicolor L.) highest cumulative crop yield (12.4
Mgha-1)
Kimetu et al. Mitigation of soil degradation with doubling of crop yield in the highly
(2008) biochar. Comparison of maize yields in degraded soils from about 3 to
degradation gradient cultivated soils in about 6 tons/ha maize grain yield
Kenya.
*source of selected references (Woolf 2008)

34
3.2.3. Soil moisture retention

In soil science the following principles are widely accepted and published (summarised in
Krull et al. 2004): The mineral and organic components of soil both contribute to soil water
holding capacity, but only the latter can be actively managed. Water is held more tightly in
small pores, so clayey soils retain more water. The lower soil bulk density generally
associated with higher soil organic matter is a partial indication of how organic matter
modifies soil structure and pore size distribution.
Many studies where the effect of biochar on crop yield has been assessed have cited
moisture retention as a key factor in the results. Given that the pore size of biochar is
relatively fixed, whereas that of the mineral soil is determined primarily by texture, it may be
expected that charcoal increases available moisture in sandy soil, has a neutral effect in
medium textured soils, and decreases available moisture in clayey soil. Any effect of biochar
particle size may be short-lived, as it appears to physically break down into fine fractions
relatively quickly (Section 3.1.5)
Experimentally, the usual technique for assessing pore size characteristics is the moisture
release curve, which indicates how quickly soil moisture is drawn from a soil under
increasing tension. The method is well suited to discriminating differences between soils of
contrasting texture, but its sensitivity may be less satisfactory for discriminating the effect of
contrasting management at one specific location: high levels of replication may be necessary
to show a significant impact of a management intervention of realistic magnitude.
In a more recent study (Gaskin, 2007), moisture release curves were determined using
samples of loamy sand soil from a field experiment where biochar had been added at rates
up to 88 t ha-1. For soils where biochar was added at rates up to 22 t ha-1 there was no
difference compared to non-amended soil, but at the highest rate the difference was
significant at water potentials in the range 0.01–0.20 MPa. At the highest potential the mean
volumetric water content impact was doubled by the biochar addition.
Soil temperature, soil cover, evaporation and evapotranspiration affect available water in soil.
Comparison of actual volumetric water content between biochar-amended and control soils
in field experiments may therefore be confounded by any indirect effect of biochar on plant
growth and soil thermal properties.
Soil organic matter increases soil water holding capacity and in the biochar-enriched terra
preta with their associated higher levels of soil organic matter, Glaser (2002) reported a
water retention capacity that was 18% higher than in adjacent soils in which charcoal was
low or absent. This was likely a combined effect of the char itself and the higher levels of
organic matter that this promotes (Section 3.1.6.4).

3.2.4. Nutrient retention and use-efficiency

There are several reasons to expect that biochar might decrease the possibility of nutrient
leaching in soils, and enhanced nutrient cycling has been cited in various field studies for
positive impacts on yield. However, very few studies have demonstrated the effect or
attempted quantitative description of the mechanism.
In general, the mineral and organic fractions of soil can both contribute to overall CEC, which
affects the ability for soils to buffer periodic flushes of ammonium that result from application
of chemical fertilisers or manures, or bursts of organic matter mineralisation during
favourable, seasonal conditions. The adsorption of ammonium ions is a relatively loose
association that does not necessarily prevent plant acquisition, yet greatly mitigates the
potential for leaching loss and the diffuse pollution issues of drinking water quality and
eutrophication of riverine and estuarine water bodies. Since considerable fossil energy is

35
required to fix nitrogen into fertiliser (Section 2.1.3.6), a low ratio of fertiliser nitrogen
application to crop nitrogen uptake can impact the overall carbon balance of agricultural
activities (see Section 3.1.6.1). Higher fertiliser use efficiency should lead to a lower fertiliser
requirement per unit yield and usually lower nitrous oxide emission (Section 3.1.6.2).
Only certain mineral constituents of soil contribute to CEC on account of abundance, and
hence surface area, and mineralogy, with certain types of clay being most important. On a
mass basis the exchange capacity of soil organic matter may be greater than for any clay
(and up to 50 times greater), but it is a relatively small proportion of soil mass in most
agricultural situations, particularly under tropical conditions. Given these factors, heavy
textured soils under climates that favour higher levels of organic matter show the highest
contributions of organic matter – about one-third – to total soil CEC (Stevenson, 1982). Since
mineralisation of organic matter is a major source of ammonium release in soil, attempts to
raise soil organic matter by increasing rates of input may not decrease – and can potentially
increase – leaching losses. In addition to the chemical stabilisation of nutrients, the physical
structure of soil determines its capacity to hold water, and hence soil nutrients in solution
(Section 3.2.3 above).
There are several reasons to expect that biochar might modify leaching potential in soils.
Available evidence suggests that on a mass basis, the intrinsic CEC of biochar is
consistently higher than that of whole soil, clays or soil organic matter. An analogy may be
drawn to the extreme CEC of activated carbon, which is relevant to its function as a sorption
medium for decolourisation and decontamination. Since secondary thermal treatment of
charcoal is one means of carbon activation, it is not surprising that the process parameters
impact the CEC of primary biochar products with temperature increasing this property
(Gaskin, 2007). This is a function of both enhanced specific surface area and the abundance
of carboxyl carbon groups that they display. The indirect affect of biochar that may result
from its modification of soil pH has not yet been included in most studies by, for example,
applying lime to the control soil.
Whilst determination of CEC and water release curves in homogeneous materials such as
biochar should be straightforward, it is more complex to quantitatively determine the
contribution of biochar once added to soil. Furthermore, the observation that CEC of biochar
may develop over time through both abiotic and biotic modification of its surfaces (Cheng et
al., 2006) implies that in order to develop a predictive, quantitative understanding, methods
to recover aged biochar from soil is required.
Information on the CEC of pyrolysis products is limited mainly by the availability of materials
produced from a sufficiently diverse range of feedstock under different production conditions.
Information on the CEC of char naturally present in soils is limited by isolation methods, so
available studies tend to rely on a comparison of whole soils amended and non-amended
with biochar (Lehmann, 2003; Liang, 2006).
The second mechanism for mitigation of leaching relates to the physical retention of soil
water, which may be enhanced by biochar in coarse-textured soils and any indirect effect of
biochar on the accumulation of soil organic matter (see previous, Section 3.2.3).
The inherent stability of biochar confers a distinction between the CEC benefits that are
possible compared to other soil organic matter; importantly there is no immediate constraint
to the level that can be attained by repeated addition, so in principal this capacity could be
incrementally enhanced. Provided that biochar is biologically stable (Section 3.1.5), the
benefit of higher CEC may be obtained without the risk of contributing to seasonal flushes of
nitrate.
The possible contribution of modified soil water dynamics and CEC to the apparent effects of
biochar on nitrous oxide emission were discussed in Section 3.1.6.2. In addition to mitigating
greenhouse gas emissions, limiting gaseous nitrogen loss can be relevant to crop fertiliser
requirement.
A beneficial impact of biochar on the plant-available phosphorus has been observed in soils
enriched with biochar, which in contrast to ammonium, is not a characteristic generally

36
associated with soil organic matter (Lehmann, 2007b; Steiner et al., 2007). In the context of
nutrient availability, the impact of biochar addition on pH may be important.

3.2.5. Use of biochar to manage water quality

Biochar may offer benefits in reducing diffuse pollution originating from agriculture through
deployment in soils from which polluting elements arise. It may also be possible to utilise its
sorptive capacity to remove contamination in the water treatment process. Studies that
demonstrate the capacity for biochar to remove nitrate (Mizuta, 2004) and phosphate
(Beaton, 1960) in this context have been cited, and in by-passing the complexity of the soil
system, controllability is achieved. However, whilst biochar may loosely hold nutrient
elements in a plant-available form, the by-product of water treatment could also be
contamination by toxic organic compounds in wastewater; biochar also has an affinity for
organic compounds (Kookana, 2006). This could confound use of the post-treatment biochar
product on land; the economic and overall carbon and environmental gain to be achieved
from centralised versus diffuse deployment for management of water quality have yet to be
assessed. The precedent for a centralised approach is the current use of activated carbon for
the removal of chlorine and organic chemicals such as phenols, polychlorinated biphenyls,
trihalomethanes, pesticides and halogenated hydrocarbons, heavy metals, and organic
contaminants (Boateng 2007). It is not clear whether the higher surface area and sorptive
capacity resulting from activation of biochar from agricultural crop wastes (Zanzi, 2001)
results in significant differences compared to biochar.

3.2.6. Potential risks to soil and water from use of biochar

Charcoal production and use appears engrained in many cultures and the apparent success
and longevity of the civilisation that created the terra preta provides some reassurance as to
the long-term safety of biochar incorporation to soil. Currently in Japan, a strong tradition in
the use of charcoal as an authorised soil improver for horticultural and agricultural
applications means that 15,000 t of carbonized material is annually applied to soil (Okimori et
al., 2003).
Nonetheless, a critical and non-prescriptive experimental analysis of risks that might arise
from the deployment of biochar has not been undertaken according to modern criteria, taking
into account all risks associated with production, distribution and physical application of
biochar, as well as its impacts in the soil. The analysis must also be based around products
of slow and fast pyrolysis, rather than simply biomass carbonisation.
This assessment is critical for three reasons: the irretrievability of biochar once added to soil,
the apparent general permanency of biochar once in the soil and the scale and speed at
which the strategy needs to be implemented to contribute to climate change mitigation. In
addition, the issue of responsibility and liability with respect to large scale application to land
is an impediment for companies seeking to invest in the production of biochar or the sale of
food products from treated land, as well as being a moral and political issue for Government
and regulatory bodies.
To date, available information is focused on the two classes of toxic compounds that are
associated most often with combustion processes, namely PAHs and dioxins. Dioxins
predominantly form at temperatures in excess of 1000°C and there are no published studies
to confirm their absence in biochar products (Garcia-Perez, 2008).
The proliferation of PAH in secondary pyrolytic reactions above 700°C is well established
(Ledesma et al., 2002), but smaller quantities may form in the temperature range of pyrolysis
reactors (Garcia-Perez, 2008). Unpublished analyses of several biochar samples also found
a PAH content no greater than that of bulk soil (Manning, pers. comm.); a single published
study examined the full PAH profile (40 individual PAH compounds) in a number of synthetic
char samples manufactured at relatively high heating rate concentrations (Brown, 2006).
Total PAH concentration was 3–16 μg g-1, depending on peak temperature, compared to
28 μg g-1 in char from a prescribed burn in pine forest. Information contained in PAH may

37
provide a measure of thermal history (Brown 2006), but empirical relationships to relate them
to process parameters have not been defined.
The above analyses determine principally the total initial content. It is not clear over what
timescale these compounds are altered in the soil, and most importantly, the bio-available
component in soil is not known. However, it has been stated by Ahmed (1989) that whilst
biochar should contain systems of PAH, existing evidence indicates that no leachable PAH is
present.
No results of bio-assays using biochar in soil have been reported, nor have the appropriate
biochar concentrations been defined in the context of the accumulations that might occur in
water and marine sediments.

4. POLICY CONTEXT AND ANALYSIS

4.1. A framework to evaluate applications of biochar

The strategies for the use of biochar considered here are those that result in biochar being
applied to soil on a significant scale. The strategies take a broad geographic perspective and
look to avoid significant practical, regulatory or economic obstacles. They therefore are those
situations where the benefits exceed the price of the biochar product; benefits may apply to
the economy as a whole, arise from economic benefits for the individual enterprise or provide
other non-monetary benefits from the use of biochar in soil. Non-monetary benefits include
the opportunity cost to a biochar producer of not utilising the pyrolysis residue in combustion
to realise its residual energy content.
However, the application of biochar to land must also be in accordance with regulatory
frameworks and law and until relevant standards are defined, the direct costs to individual
users in addressing these controls may be prohibitive. Although scenarios might involve
large scale bioenergy and industrial agriculture, the same framework should be used to
evaluate its potential contribution to subsistence or slash-and-burn agriculture. However,
although these practices contrast quite starkly, like soil and climatic factors, farming practices
occur over a continuum of different scales, and these examples sit at opposite extreme ends.
As such, it will be highly advantageous to define a single framework for assessment and
comparison of different biochar scenarios for their net carbon benefit and socio-economic
impacts.
Separate evaluations should be made for the economic and environmental sustainability of
alternative biochar scenarios. If the assured carbon-equivalent gain available using biochar is
positive but the economic analysis for mainstream agriculture negative, then utilisation of
economic instruments – most likely carbon trading or a subsidy that ensures biochar is used
in soil rather than for combustion – is essential. The introduction, expansion or revision of
such instruments that place a monetary value on the utilisation or disposal of organic waste,
maintenance of soil quality and support for renewable and bioenergy as a whole may then be
considered.
For any biochar scenario it is possible that the agronomic value for biochar is sufficient to
render the economic evaluation positive, without resorting to carbon markets or Government
incentives. Then concerted research effort will be sufficient to establish certainty around the
extent and realisation of such benefits.

4.2. Scenarios for the uptake of biochar for use in soil

Even pending further research, biochar may be attractive to producers of high value crops,
where certain characteristics of biochar (such as water storage) have high economic value.
In these markets the price of biochar may be acceptable even in the absence of subsidy or
payment. Some additional brand value may be derived from the carbon balance of the
production system but would not be the driver of the system. Biochar could also be profitably

38
employed on recreational land or on sports turf. Such applications may increase recognition,
but will not provide the extent of use required to contribute to climate change mitigation.
Currently, uncertainty around the expected benefits and potential returns at current prices is
likely to limit widespread use in mainstream agriculture. This uncertainty stems from
inadequate understanding or quantitative description of the underlying processes and the
multiplicity of potential benefits and interactions.
Key biochar characteristics will vary according to the nature of the agricultural management
system, soil and climate, and may not be static over time. The number of useful biochar
properties and their relative importance will vary accordingly, but at the moment the
understanding necessary to produce biochar optimised to deliver a particular balance of
properties – especially in tandem with viable energy capture – does not yet exist. The
complexity and diversity of decision making on farms, and the susceptibility of soil
management strategies to commodity prices and external economic forces is important too.
To date, assessments of the benefits to be derived from applying biochar to soil have been
made on the basis of very limited experimental evidence, and are often scoping studies
based on generalised situations. The sensitivity of proposed scenarios to the spatial
dimension presented by climate is improving but still inadequate in predictive terms.
Comprehensive, whole-system life-cycle analyses (LCA) with full accounting are required in
order to avoid unintended negative consequences. In such analysis it will be recognised that
deliberate accentuation of one biochar characteristic may impact on the delivery of others, for
example porosity versus nutrient value. Thus not only does the predictive capacity for
biochar ‘performance’ not currently exist, but the feasibility of optimising multiple useful
characteristics is not known. This is inhibiting realisation of other benefits to the wider
system.
In the financial evaluation of biochar technologies it is important to consider not only current
prices, which are known or can be determined, but also their likely future value. Given the
current trajectory of global greenhouse gas emissions, the price of emissions as a tradable
commodity will be increasingly important. The future price of fossil fuel and subsidy levels for
renewable energy are difficult to predict. However, the indirect gains from the use of biochar
will probably increase with the future price of fossil energy, reflecting the generally energy-
intensive nature of the key inputs associated in mechanised agriculture, and possibly with
finite supply of water (irrigation costs). There will be a corresponding increase in value of the
residual energy in biochar as fossil energy prices rise. The price of bioenergy may also be
enhanced by Government subsides designed to improve energy security and promote
environmental goals. In many industrialised countries the opportunity cost associated with
using biochar in soil is artificially enhanced by renewable energy subsidies. The future price
of grains and other commodities is difficult to predict, but changing diet, a growing global
population, and increasingly limited supply of new agricultural land is likely to increase
demand relative to supply.

4.3. Market intervention and carbon trading

Markets for the sale of pyrolysis feedstocks are not currently accessible, and markets for
potential feedstocks are ill-developed. A market for credits relating to ‘avoided emissions’ in
which land managers could engage does not yet exist. In general, there also remains a lack
of knowledge and awareness of bioenergy and carbon markets, how to access these
markets, and particularly a way to accurately evaluate costs and benefits associated with the
use of biochar in soil.
In the absence of research to support the optimisation of biochar and its agronomic
evaluation, the viability of biochar-based soil management based on carbon-offsets alone is
important. However, no framework exists within which the carbon sequestered in biochar can
be certified as a tradable commodity. This barrier extends beyond carbon trading under the
UN Clean Development Mechanism (CDM) to the voluntary carbon markets. To date the
methodology required to recognise the stabilisation of degradable organic matter as an

39
‘avoided emission’ of CO2 has not been presented, although this is a current activity within
the International Biochar Initiative.
Additional evidence for the portion of biochar that can be considered stable over the long
term is required. Furthermore, research is required to determine whether the carbon-
equivalent benefits of biochar application to soil extend beyond direct carbon sequestration
or the avoidance of other greenhouse gas emissions. At the moment these effects are not
fully understood or proven in the predictive sense, nor have the timescales over which they
mainly occur been established (Section 3.1.5.). However, the existence of a current
methodology for stabilisation of organic matter in avoidance of methane emission represents
an important precedent (UNFCCC).
Various national Governments have implemented carbon trading schemes in order to meet
emission reduction commitments under the Kyoto protocol. This has resulted in the
European Union Greenhouse Gas Emission Trading Scheme and subsidiary schemes such
as the UK Emissions Trading Scheme. More recently, Australia has announced plans for a
Carbon Pollution Reduction Scheme (Anon, 2007c).
In these countries, their regional neighbours and in the USA there is a strong market for
voluntary carbon trading carbon. Individuals or organisations engage outside of Government
requirements in a market currently valued in excess of USD 30 billion yr-1. Offsets traded in
the voluntary market are unregulated, and their credentials have been questioned. Biochar-
based schemes should offer the opportunity to trade a more demonstrable offset (by virtue of
the visibility and durability of biochar in soil), that has community as well as climate benefits
through its impact on agriculture. However, there is an expectation within some NGOs that
the inclusion of biochar into any carbon trading scheme will ultimately lead to large projects
based around dedicated biomass crops that will not benefit agriculture and put additional
pressure on rural livelihoods in developing countries as a consequence of land acquisition.
Other major NGOs have not yet adopted a specific position on biochar.
Alongside the IBI, pressure groups such as the US-based Clean Air Task Force have been
promoting biochar based offsets alongside other bioenergy schemes for trading in all carbon
markets (Baum et al., 2006).

4.4. Market acceptability issues

There are significant organisational and institutional obstacles to the use of biochar in soil.
Since biochar could be used on a wide scale and cannot be removed from soil once applied,
there is a need to carefully assess any potential negatives in occupational health (possible
inhalation risks from physical application to soil), environmental pollution (particulates
travelling into water or air), water quality (impacts on aquatic life and water treatment) and
food safety (surface and systemic contamination of food products). Since several sectors are
involved – broadly water, waste and food safety – it requires a concerted effort to evaluate
potential products, and ideally define product standards. Support for the use of biochar in
meeting policy objectives will draw upon life-cycle analysis with full greenhouse accounting,
backed by a body of experimental data. Where biochar is designated as a regulated waste
material, land-users in many countries may be subject to a complex and expensive approval
process pending defined standards.
However, the political and economic case for using food crops as feedstock for liquid biofuel
production (bioethanol), or devoting large land areas to biomass production for bioenergy, is
increasingly challenged in the context of rising commodity prices and increasing land
pressure. Strategies based around pyrolysis with the use of biochar in soil are distinct from
these, since productivity and sustainability of land is potentially enhanced. Globally, rising
prices favour cash-crop farming, and the economic case for applying biochar to land is likely
to improve.
The lack of mechanistic understanding as to the function of biochar and its interaction with
already complex soil processes, mean predicting the return to an investment in biochar
between locations in terms of extent, predictability and durability of benefits does not yet

40
exist. Providing a measure of certainty to the many possible benefits is a key challenge to be
addressed by further research.

4.5. Research
To date there are a limited number of examples of large-scale publicly funded research
initiatives that assess the use and optimisation of biochar for use in soil. In New Zealand the
Massey University has a Biochar Research Initiative. In the UK the Engineering and Physical
Sciences Research Council supports two established projects, one building capacity in
technology for fast pyrolysis (as part of a wider bioenergy initiative, SUPERGEN) and testing
by-products in soil, and the other developing Carbon Sequestration and Capture technology.
Brazil has funded second and third tier levels within ‘macro-programs’ defined by the
research organisation, EMBRAPA. These will extend field experimentation to create new
terra preta (terra preta nova). At Federal Government level, the US has created the Farm Bill
that supports ‘biochar research development and demonstration’ which seeks to enhance
agricultural energy programs (Anon, 2007b). Biochar was specifically mentioned in the
Garnaut Climate Change Review in Australia (Garnaut, 2008) and has been raised in
Environment Select Committee discussions in the UK.
Currently much of our understanding of the long-term dynamics of biochar is based on
studies of charcoal from natural fire, new charcoal produced using traditional methods or
analogous procedures undertaken in the laboratory. Studies in Brazil use charcoal fines
(waste) from industrially produced charcoal. However, only a small number of
comprehensive studies using the products of commercial bioenergy plants currently exist,
and although the conditions used to produce these products has been guided by preliminary
studies, those conditions may not have been optimised for the soil into which they have been
incorporated. The only full-scale field trials using biochar from slow pyrolysis energy plants
are being conducted in NY, USA
(<http://www.css.cornell.edu/faculty/lehmann/research/biochar/biocharproject.html>).
.

5. RESEARCH PRIORITIES AND FUTURE CHALLENGES

Based on the results of this review, the following research priorities have been identified:
1) Determine a predictive relationship for properties and qualities of biochar and its
manufacture such that it can be optimised for use in soil.
2) Examine how the possibility of adverse impacts on the soil and atmosphere can be
eliminated with certainty.
3) Model the impact of alternate bioenergy systems on the carbon cycle at the global scale,
and in the context of national targets, in order to support policy decisions and devise
suitable market instruments.
Since the underlying context for biochar-based strategies is that of global climate change,
research needs to provide answers that are applicable under diverse combinations of
climate, agriculture and energy production systems. This requires a fundamental,
mechanistic understanding of how biochar provides its unique functional characteristics,
probably embodied in models, and would include its interactions with other living and non-
living components of soil.
Globally coordinated research activity across a range of countries and climates is necessary
if the global applicability of knowledge gained is to be rigorously assessed.

41
5.1. Fundamental mechanisms
The fundamental mechanisms by which biochar could provide beneficial function to soil and
the wider function of the agro-ecosystem are poorly described in terms of providing the
predictive capacity that is required.
In short-term experiments of months to a few years, biochar addition seems to generally
enhance plant growth and soil nutrient status and decrease N2O emissions. Yet surprisingly
little is yet published concerning how these benefits occur, or particularly why the effects are
quantitatively so variable according to crop, soil and application rate. Soil-biochar dynamics
need to be investigated. The required understanding will have to be obtained in the following
areas:

a) Functional interactions with soil microbial communities. Biochar may modify the
symbiotic relationships that exist in the rhizosphere, that is between plants and microbes
in close proximity to the root. At the moment the net effect of physical protection provided
to microbial colonies, and adequate access of the same colonies to labile and soluble
carbon substrates is not yet known. The component of biochar stability provided by
association of individual biochar particles and fine mineral particles has not been
established, and the role of microbial and rhizosphere secretions in promoting them will be
important. Fundamentally, the apparent conflict between high stability, soil organic matter
accumulation and apparent enhancement of soil microbial activity needs to be resolved.
Useful methods will separate indirect effects of increased water holding capacity or altered
water release characteristics, pH effects, and allow for their potentially transitory nature.
b) Surface interactions. It seems that as the exchange capacity of biochar surfaces
develops over time, the contribution of feedstock and production parameters to the
trajectory of its development will need to be established. Once the relative importance of
biotic and abiotic processes in promoting this development are known, and the net effect
of any simultaneous change in the ratio of external- to internal-surface resulting from
physical disintegration under soil movement quantified, the net effect of climate might be
predicted. Interaction of biochar with anions, most importantly phosphate, needs to be
established, and the extent to which nutrient effects are internal, i.e. derived from within
the biochar (finite), and external, supplied by the wider soil, must be determined.
c) Nutrient use efficiency. Understanding the link between biochar function and its
interaction with nutrient elements and crop roots may enable fertiliser use efficiency to be
enhanced and diffuse pollution of watercourses and wetlands.
d) Soil physical effects. The intrinsic contribution that biochar can make to the wetability of
soil, water infiltration, water retention, macro-aggregation and soil stability is poorly
understood – yet should be of critical importance in tropical environments in combating
erosion, mitigating drought and nutrient loss, and in general to enhance groundwater
quality. The loss of biochar through vertical or lateral flow is not well understood; only
recently have studies been initiated to examine movement down the soil profile.
e) Fate of biochar. The stability of biochar carbon is intrinsic to fulfilling its role as a
significant CO2 sink, but in order to perform an agronomic role, it must also remain within
the soil to which it is applied. The environmental role or impact of biochar once it has
moved through a soil profile, or into watercourses, is yet to be assessed. Information on
the extent to which physical breakdown of biochar changes the balance in its properties,
particularly with respect to soil water dynamics, exchange capacity and soil micro-and
macro-aggregation is lacking. Methods are urgently required to assess the long-term
biological stability of specific biochar samples, possibly extrapolating from the dynamics of
atypically high initial rates of decomposition.
f) Impacts on soil N2O and CH4 emission. Published data for the effect on trace gas
emission is extremely limited, but has a potentially great impact on the net benefit of a
biochar strategy. Good predictive models will be necessary for this to be reflected in future

42
 accounting for biochar projects, and at the moment there is not a clear mechanism for the
effect.
g) Plant physiological responses to biochar ultimately dictate impacts on growth and yield
and need to be directly targeted in future work.

5.2. Properties, qualities and environmental risk assessment

As the mechanisms underlying biochar function in soil become understood, biochar products
can be optimised to deliver specific benefits using a defined screening procedure based on
relevant properties. Research tools for comparing different biochar products already exist
from at least two decades of work on charcoal and other black carbon in soils: 13C nuclear
magnetic resonance spectroscopy has proved perhaps the most useful in revealing gross
differences in composition at the functional group level. However, a screening approach
should be defined by relevant properties rather than available measurement techniques. Also
a cheap and rapid method procedure appropriate to routine analysis has not been identified.
The following research is needed:
a) Scoping. A comprehensive and systematic analysis of commercially available biochar
products, including charcoal produced by industrial- and farm-scale carbonisation
processes, to scope the boundaries of variation in compositional and functional
characteristics of biochar. This should utilise all available techniques and link to a
database recording the type of biomass from which the biochar was produced, and the
type and details of the production process for correlative analysis.
b) Benefits and risks. Thorough environmental and agronomic evaluation of biochar in soil
will enhance its acceptability as a large-scale offsetting and sequestration strategy for
CO2. However, the incorporation of biochar into soil is irreversible and therefore must be
safe, with biochar products free of potentially harmful contaminants.
a) Inventory. A systematic screening of biochar products is required, in tandem with the
above, to establish the presence and absence of phytotoxic materials that could
conceivably be formed during pyrolysis. This will establish the range of potentially
harmful chemical contaminants present and their peak concentrations, providing
evidence essential to the preparation of risk assessments. Key compounds will be
polycyclic aromatic hydrocarbons, established products of partial combustion, and
residual oils and acids.
b) Air pollution. The environmental impact of vapours and gases produced in open
combustion associated with traditional charcoal production and in sub-surface
combustion, needs to be carefully evaluated, along with the technological developments
required to address it. These emissions can cancel out carbon sequestration if not
contained or used in the process.
c) Waste pyrolysis. The potential for urban waste to be used in biochar production needs
to be assessed. A risk-based approach may dictate that such materials are more
suitable for gasification than pyrolysis if harmful compounds are abundant. It is not
known whether the low quality biochar produced from these waste streams is suitable to
deliver sequestration and soil benefits.
d) Indirect impacts. The implications of rapid expansion in biomass pyrolysis on
agricultural and natural land areas is a concern through, for example, the expansion of
fast-growing plantation forest for production of feedstocks for biochar or charcoal
production. There is also the possibility of increased deforestation if the technology is
allowed to expand in an uncontrolled way.
e) Scrubbing air pollutants. Biochar has been reported to scrub CO2, nitrous oxides and
sulphur dioxide from fuel gas, creating a nitrogen-rich biochar product that could
substitute conventionally produced chemical fertiliser. Since this approach addresses the
practicality of application and offers added benefits, it needs to be pursued and refined.

43
5.3. Carbon cycle modelling
Two types of carbon modelling are required: static spreadsheet models to compare
alternative scenarios for their relative carbon-equivalent gain, and mechanistic soil simulation
models that capture information from short term experiments to predict longer-term impacts
on soil function. In addition, socio-economic models that incorporate a spatial dimension are
required to assess the workability of particular scenarios.
a) Modelling net carbon gain. So far only generic, theoretical analyses have been
published. Full assessment spreadsheet models based on improved experimental
evidence are required to conduct ‘what-if’ comparisons of alternative strategies for
specific feedstock streams and pyrolysis facilities, taking the spatial dimension of
feedstock supply and biochar use into account.
b) Mechanistic soil modelling. Modelling of the linked carbon and nitrogen cycles in soil
with and without intervention using biochar is essential to understanding the fundamental
mechanisms referred to above, and the impact on soil-based emissions of greenhouse
gases. Modelling of soil carbon currently relies on conceptual pools, and essentially
ignores black carbon from a mechanistic perspective. Progress in this area is dependent
on improved quantification methods for biochar in soil.
c) Economic models. Conceptual and actual geographic boundaries must be carefully set
when assessing a particular scenario, accounting for the entire supply chain. Socio-
economic constraints relevant to the application of biochar must be recognised.
d) Audit. A standard methodology for validation and audit of biochar application is required
that ideally permits the source of a particular biochar application to be confirmed
retrospectively.
e) Databases. International support for a global system that enables optimal biochar
products to be selected for application in a particular location and system. The only such
database initiated to date is CharDB, at Terra Carbona
(<http://terracarbona.org/chardb/>).

5.4. Beneficiaries
Pyrolysis enterprises. In areas where biochar may be produced for agricultural or
environmental gain rather than energy production, NGOs may be interested in exploring the
pyrolysis biochar technology. Market development would facilitate the dialogue between
producers and investors as well as researchers and users. Currently the amount of biochar
available for use as a soil amendment (and hence carbon sequestration) is limited to an
extent where even assessment of products for non-energy use is limited.
Charcoal producers. Traditional producers may experience expansion in the market for
charcoal if its use as a soil amendment is supported by land-users for carbon sequestration
or enhancing soil fertility. There is a precedent for charcoal being produced on a commercial
scale within Europe, with almond charcoal supplied to power companies in Spain.
Water companies. Larger scale production of activated charcoal from pyrolysis-derived
biochar could reduce costs. Large scale use of biochar on agricultural land in intensively
farmed areas may also reduce diffuse pollution and the need, and hence costs, for treatment
of water.
Land users. Individual farms or farming consortia would benefit due to greater profitability as
a result of savings on energy and fertilisers. Remediation of degraded or contaminated land
using biochar could be supported through Government, engaging the agricultural community
and supported by environmentally-oriented incentives and subsidy.

44
5.5. Commentary on likely barriers to the adoption of a large scale
enterprises utilising Biochar
If the technical limitations described above are addressed and if biochar-related techniques
are judged to be viable methods to address climate and agricultural problems, then it is likely
that additional barriers to widespread adoption will remain. This section provides a
commentary on these barriers.
The economics of biochar production will be fundamentally altered once methodology has
been proposed and adopted with regard to carbon offset values, in the voluntary market, and
as a longer-term possibility, certification under the United Nations Framework Convention on
Climate Change (UNFCCC) clean development mechanism (CDM).
A streamlined regulatory framework may be essential to avoid stifling initial interest in the use
of biochar products in mainstream agriculture. Government policies to help ensure continuity
of feedstock supply to pyrolysis enterprises could assist in establishment of the industry;
subsidy arrangements could be adopted that favour rather than discriminate against the use
of biochar in soil.
A routine standard method to quantify biochar in soil is essential to realise the research
agenda. Supply of biochar material from commercial pyrolysis facilities is currently extremely
limited and localised, inhibiting research activity. Biochar is highly heterogeneous, so
standards and quality control need to be defined and certified.
Association of pyrolysis with wider bioenergy technology and specifically the biofuel debate
presents an image problem in the wake of diminished global grain stocks and rising
commodity prices. However, this could also present forums where the principle and multiple
benefits of biochar-based strategies can be discriminated and promoted.
The multi-disciplinary nature of the biochar concept seems to inhibit large-scale funding of
the extensive research agenda, particularly the large sums required for long term trials. It
also appears that assigning responsibility for researching biochar within Government and
between public and private sectors is challenging. Funding for testing commercial products
may be borne by companies producing the products, but such activities will need to be
integrated with public sector science to address the wider climate change agenda.

45
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World Journal of Agricultural Sciences 3 (5): 572-575, 2007
ISSN 1817-3047
© IDOSI Publications, 2007

Response of Broiler Chickens to Wood Charcoal and Vegetable Oil Based Diets

A.A. Odunsi, T.O. Oladele, A.O. Olaiya and O.S. Onifade

Department of Animal Production and Health,

Ladoke Akintola University of Technology, P.M.B. 4000, Ogbomoso, Nigeria

Abstract: An experiment was carried out to determine whether Wood Charcoal (WC) would affect growth
performance, carcass characteristics and blood profiles of broilers and to determine whether Vegetable Oil (VO)
supplementation would influence similar parameters in broiler chickens fed WC. Dietary WC was incorporated
into broiler starter and finisher diets at 0, 2.5, 5.0 and 7.5% levels while diets containing 2.5 and 5.0%WC were
each supplemented with 1.5%VO to make six dietary treatments. Results showed that feed intake (F1) was
significantly increased in broilers fed 5% WC with or without VO while birds on other diets had similar (p>0.05)
intake. Body Weight Gain (BWG) and feed conversion ratio (FCR) were significantly better (p<0.05) on birds
fed without WC compared to those fed on WC based diets. Supplementation with VO only gave marginal
improvement on performance indices when compared with the corresponding WC based diets without VO.
However, the performance criteria used (F1, BWG and FCR) were still not commensurate to the control diet.
Carcass yield, liver, spleen and kidney did not show any major variations (p>0.05) among dietary treatments
while lung, heart and gizzard indicated significant differences (p<0.05). The packed cell volume, haemoglobin,
red blood cell and white blood cell, MCV, MCH, MCHC values obtained for the six treatments were not
statistically (p>0.05) different from each other. The present study appeared not to justify the dietary inclusion
of WC in broiler chicken diets and its use in broiler diets is not recommended.

Key words: Wood charcoal % vegetable oil % broilers % growth performance % carcass yield % hematology

INTRODUCTION whether Vegetable Oil (VO) supplementation would affect

In Nigeria and indeed many other countries, various
feeds and additives are incorporated into poultry diets to
ensure maximum productivity. Most of the additives are
used depending on area and the ease of use. Moreover,
most of these materials are not cited in the scientific
literature but are used locally, for instance wood charcoal
[4, 6]. It was reported by Kutlu et al. [4] that some local
poultry producers in Turkey claimed that 20-50 g wood
charcoal per kg diet prevents fatness and improve
performance of broilers and layers. In view of these
assertions, we have attempted to validate this claim in our
laboratory because wood charcoal is widely available in
Nigeria and so far, no reports can be cited as to its
nutritional use apart from being a suitable and alternative
cheaper source of generating heat.
The present study was therefore conceived to
determine whether dietary wood charcoal applied at
graded levels would influence growth performance,
hematology and carcass characteristics and to determine
similar parameters in broiler chicken fed wood charcoal.
MATERIALS AND METHOD
Experimental diets and their composition: Wood
Charcoal (WC) was obtained from a local market in
Ogbomoso and ground through a mill to pass a 1mm
sieve. As ground, it contained 946g DM/Kg, 154g ash/kg,
97.5g crude fibre/kg, 10.8g ether extract/kg, 19.6g crude
protein/kg and 664.1 g nitrogen free extracts/kg. Six diets
each were formulated during the starter (1-5 weeks) and
finisher (5-9 weeks) phases. Diet I was designated as the
control without WC while diets 2, 3 and 4 contained 2.5,
5.0 and 7.5% WC respectively. Diets 5 and 6 were
formulated to contain 2.5 and 5.0% WC respectively each
supplemented with 1.5% Vegetable Oil (VO). The diet
composition for the two phases is shown in Table 1.
Experimental birds and management: A total of 200
unsexed Anak 2000 broiler chicks were procured from

Corresponding Author: Dr. A.A. Odunsi, Department of Animal Production and Health, Ladoke Akintola University of
Technology, P.M.B. 4000, Ogbomoso, Nigeria
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 World J. Agric. Sci., 3 (5): 572-575, 2007

Table 1: Composition and nutrient contents of broiler starter and finisher diets (%)
Starter diets Finisher diets
---------------------------------------------------------------------------------- ----------------------------------------------------------------------------------
1 2 3 4 5 6 1 2 3 4 5 6
Ingredients 0 2.5 5.0 7.5 2.5+VO 5.0+VO 0 2.5 5.0 7.5 2.5+VO 5+VO
Maize 52.7 49.7 43.7 47.7 47.8 44.9 58.4 55.4 53.4 49.4 52.5 49.5
Groundnut cake 30.6 31.1 31.6 32.1 31.5 32.0 25.4 25.9 26.4 26.9 27.3 27.8
Wood charcoala 0.0 2.5 5.0 7.5 2.5 5.0 - 2.5 5.0 7.5 2.5 5.0
Vegetable oil - - - - 1.5 1.5 - - - - 1.5 1.5
Fish meal 5 5 5 5 5 5 2.5 2.5 2.5 2.5 2.5 2.5
Wheat offal 8 8 8 8 8 8 10 10 10 10 10 10
Bone meal 2 2 2 2 2 2 2 2 2 2 2 2
Oyster shell 1 1 1 1 1 1 1 1 1 1 1 1
Salt 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25
Methionine 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2
Premix 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25
Analyses
Dry matter 91.8 91.8 91.8 91.8 91.7 91.8 91.3 91.5 91.6 91.3 91.5 91.2
Crude protein 22.9 22.8 22.9 22.9 22.9 23.0 19.3 19.4 19.6 19.5 19.4 19.5
Ether extract 8.79 8.73 8.66 8.59 8.68 8.57 5.64 5.35 5.47 5.31 6.38 6.74
Crude fibre 3.46 3.50 3.52 3.56 3.46 3.49 4.84 4.05 4.25 4.65 4.28 4.15
Crude ash 3.47 3.66 3.86 4.05 3.67 3.86 2.85 2.96 2.39 3.14 2.94 2.59
NFE 53.2 53.1 52.9 52.7 53.0 52.9 59.5 59.8 58.6 58.6 58.4 58.2
Wood charcoal contains ( %); DM, 94.6, CP, 1.96; EE, 1.08; CF, 9.75; Ash, 15.4, NFE, 64.41
a

Farm Support Services, Ibadan and fed a commercial
broiler starters mash (24%CP/2900 ME kcal/kg) for 1 week.
Subsequently, one hundred and eighty birds were
weighed and randomly allotted to the six dietary
treatments in triplicate lots of 10 chicks each using the
completely randomized design.
The six experimental starter and finisher diets were
provided to the birds during the starter (1-5 weeks) and
finisher (5-9 weeks) phases respectively. The groups were
kept in a floor-littered poultry house situated at the
University Teaching and Research farm, Ogbomoso. Feed
and water were provided ad libitum. Other routine
management practices such as vaccination, drug
administration and maintenance of cleanliness in and out
of the poultry house were applied.
Initial body weights of the birds were taken on
replicate basis at the start of the study and thereafter on
weekly basis. Weekly feed intake was also recorded. The
mean daily weight gain, daily feed intake and feed to gain
ratio were thus calculated from the data obtained during
the starter, finisher and overall experimental period. On
day 63, 2 birds of mean weight close to the average group
weight were randomly selected from each of the 18
replicates and starved of feed for 12 hours in order to
empty their crops. The birds were exsanguinated,
defeathered, eviscerated and dressed. Each bird’s carcass,
cut-up parts and organs were separately weighed and
expressed as a percentage of dressed weight. Blood
samples were collected on the 63rd day of the trial from 3
birds per treatment during slaughter. The samples were
collected in bottles containing ethylene tetra-acetic acid
(EDTA) as anticoagulant. They were then taken to the
laboratory for hematological analyses that included
Packed Cell Volume (PCV), erythrocyte (red blood cell),
leucocytes (white blood cell) and haemoglobin. The Mean
Cell Volume (MCV), Mean Cell Haemoglobin (MCH) and
Mean Cell Haemoglobin Concentration (MCHC) were
calculated. The hematological parameters were determined
as described by Davice and Lewis [2].
The proximate compositions of wood charcoal
and experimental diets were determined according to
AOAC [1]. Data collected were analyzed by analysis of
variance technique and the Duncan’s multiple range
technique was used to detect differences among treatment
means [8].
RESULTS AND DISCUSSION
The growth performance data of broiler chickens fed
wood charcoal based diets supplemented with or without
vegetable oil during the starter, finisher and overall
experimental periods is shown in Table 2. At the starter

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 World J. Agric. Sci., 3 (5): 572-575, 2007

Table 2: Effect of providing wood charcoal and supplemental VO on growth, performance of broilers at starter, finisher and the overall experimental period
Variable Control 2.5%WC 5%WC 7.5%WC 2.5%WC+VO 5.0%WC*VO SEM
Days 7-35
BWG (g/b) 20.5a 19.5a 22.4a 17.3b 22.2a 21.4a 0.95
Feed intake (g/b) 48.6b 50.9b 54.0a 49.0b 50.5b 51.0b 1.44
FCR(F1/BWG) 2.37c 2.61b 2.41c 2.84a 2.73b 2.38c 0.46
Days 35-63
BWG 50.3a 44.5b 42.6b 41.8b 44.7b 44.5b 1.30
Feed intake 129.2b 127.9b 135.4b 122.4c 127.4b 132.3a 2.55
FCR 2.57c 2.87b 3.18a 2.93a 2.84b 2.97a 0.12
Days 7-63
BWG 35.3a 32.0b 32.5b 29.6b 33.5b 33.0b 0.60
Feed intake 88.8b 89.4b 94.7a 85.7c 88.8b 93.2a 2.06
FCR 2.52b 2.791b 2.91a 2.90a 2.65c 2.68c 0.07
a, b, c: Means within a row with similar superscripts are not significantly different (p>0.05)

Table 3: Effect of dietary charcoal wood on carcass weight, carcass yield, cut-up parts and organ weights of broilers
Diets
----------------------------------------------------------------------------------------------------------------------------------------------------------
0%WC 2.5%WC 5%WC 7.5%WC 2.5%WC+P0 5%WC+P0
Variable 1 2 3 4 5 6 SEM
Carcass
Carcass weight (g/b) 1445.3a 1260.4b 1303.1ab 1155.3c 1470.8a 1387.9a 28.02
Carcass yield (% ) 73.1a 70.3a 71.6a 69.7b 78.4a 75.1a 0.34
Abdominal fat (%) 0.95a 0.65b 0.61b 0.54c 0.69b 0.71b 0.09
Relative cut-up parts (% of CW)
Wing 12.7 12.6 11.4 11.5 11.8 11.5 0.79
Thigh 19.2 16.7 22.9 19.2 19.0 17.1 0.30
Drum stick 14.6 13.7 14.8 15.1 14.7 15.7 0.47
Breast 25.3 25.3 27.6 25.1 24.4 24.5 0.35
Back 18.9 21.4 19.4 19.2 19.9 20.8 0.24
Neck 6.96 6.96 6.90 7.03 7.89 7.07 0.07
Relative organ weights (% of CW)
Liver 2.17 2.3 2.23 2.46 1.95 2.41 0.07
Kidney 0.51 0.97 1.0 0.98 0.83 0.96 0.04
Lung 0.54 0.74 0.66 0.74 0.73 0.73 0.02
Heart 0.54 0.57 0.47 0.53 0.42 0.50 0.01
Spleen 0.12 0.11 0.11 0.23 0.07 0.11 0.12
Gizzard 2.72 3.30 2.39 2.80 2.60 2.51 0.72

phase, feed intake was highest (p<0.05) for broilers on 5%
WC diet while other treatments had similar intake. Body
weight gain was similar across the dietary groups with the
exception of broilers fed 7.5% WC based diets. Feed/gain
ratio was significantly (p<0.05) better for birds fed on the
control diet, 5% WC and 5% WC+VO diets compared to
the other dietary groups however, broilers fed on 7.5%
WC had the worst feed conversion ratio.
During the finisher phase, birds fed the control diet
significantly (p<0.05) gave the highest BWG. Broilers on
5% WC with or without VO had the highest (p<0.05) feed
consumption while the least intake was recorded for birds
fed 7.5% WC diet. The control diet exhibited the best-feed
conversion ratio.
Considering the overall feeding period (combined
starter/finisher phases), feed intake was significantly
increased in broilers fed 5% WC with or without VO
while birds fed 7.5%WC diet had the least (p<0.05)
consumption. Body Weight Gain (BWG) and Feed
Conversion Ratio (FCR) were significantly better
(P<0.05) on birds fed the control diet compared to those
fed WC based diets without VO. The increase in feed
consumption for birds fed 5%WC did not really translate
to higher weight gain. However, the slight improvement in
weight gain observed with the addition of VO is a
reflection of the positive attributes of vegetable oil in
poultry diets. Vegetable oil increases energy density,
reduces dustiness and increase vitamin A concentration
in diets [7, 9].
Charcoal supplementation was reported to induce a
small reduction in feed intake, egg production and feed
conversion ratio [4]. The reduction in feed intake was

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 World J. Agric. Sci., 3 (5): 572-575, 2007

Table 4: Effect of dietary wood charcoal and supplemental vegetable oil on hematology of broilers
Diets
----------------------------------------------------------------------------------------------------------------------------------------
Variables 1 2 3 4 5 6 SEM
Packed cell volume (%) 29.00 26.50 25.50 24.50 26.00 25.00 0.24
Haemoglobin (gm %) 9.67 8.83 8.50 8.17 8.60 8.33 0.124
Red blood cell x106 4.83 4.42 4.25 4.05 4.30 4.15 0.58
White blood cell x103 8.83 9.03 9.95 9.60 6.23 6.35 0.57
Mean cell volume 59.90 60.00 60.00 60.60 60.50 60.40 0.58
Mean cell Haemoglobin 3.33 3.33 3.33 3.33 3.31 3.33 0.003
Mean cellHaemoglobin
concentrations 199.90 199.90 200.00 201.80 200.00 201.20 1.89

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with the use of WC or supplemental VO in their diets. performance of Big 6 heavy tom turkeys. J. Anim.
However, the lung, heart and gizzard showed slight Feed Sci., 11: 135-141.
(p>0.05) changes among the dietary groups. These 7. Odunsi, A.A. and A.A. Onifade, 1998. Effect of zinc
showed that wood charcoal used in this study had no bacitracin supplementation of broiler chick diets
major physiological effects on tissue or organ containing a low or high vegetable oil concentration
development and functions. However a positive in the tropics. Trop. Vet., 16: 51-57.
development is the reduction in the abdominal fat 8. Steel, R.G.D. and J.H. Torrie, 1980. Principles and
deposition in broilers fed WC based diets relative to the procedures of statistics. A biometrical approach. 2nd
control group. edition McGraw Hill Books Co., New York, USA
This pilot study reported here demonstrated that the 9. Zollitscha, W., W. Knausa, A. Aichinegera and F.
wood charcoal incorporated into broiler chicken diet did Lettnera, 1997. Effects of different dietary fat sources
adversely affect broiler performance during the entire on performance and carcass characteristics of
feeding period as opposed to reports by Kutlu et al. [4] broilers. Anim. Feed. Sci. Technol., 66: 283-287.

575
Majewska T., Pyrek D., Faruga A.

T. Majewska, Department of Poultry Science, University of

Warmia and Mazury in Olsztyn, Oczapowskiego 5, 10-718
Olsztyn, Poland

A note on the effect of charcoal supplementation on the

performance of Big 6 heavy tom turkeys

The experiment was conducted on 204 Big 6 heavy tom

turkeys. One-day-old chicks were allocated to two feeding
groups, each with three replicates of 34 birds. All of the birds
were fed identical granulated standard feeds in a three-stage
system. The control birds were fed unsupplemented feed,
the birds in the treatment group received a feed
supplemented with pulverized hardwood charcoal at a dose
of 3 kg/ton. Charcoal was given from day one of life for the
entire period of rearing. The use of charcoal had a beneficial
effect on performance. After 18 weeks of rearing, turkeys
given charcoal-supplemented feed were 5.9% heavier (on
average 870 g) and had a 6.5% better feed conversion ratio
than the control birds. Survival in the group receiving
charcoal was 99% as compared with 87.3% in the control
group. The crude protein content of the breast muscles of
the experimental group increased significantly. The
European Production Index equaled 393 for the control
group and 504 for the charcoal-supplemented group.

Journal of Animal and Feed Sciences 2002, vol: 11,

number: 1, pages: 135-141
Majewska T., Zaborowski M.

Charcoal in the nutrition of broiler chickens

The aim of the research was to define the influence of

charcoal added to the standard feed mixtures during their
production or directly before feeding broiler chickens on the
latter’s development. The experiment was conducted on 180
Starbro chickens. One-day-old chicks were allocated into 3
feeding groups, with two replicates of 30 birds each. The
birds of all groups were fed identical standard mixtures in a
three-stage system: Starter, Grower and Finisher. The
control birds were fed only with standard mixtures. The birds
in group 2 received the same mixtures but supplemented
with pulverized hard-wood charcoal at a dose of 3 kg/ton
(together 100.3%), added at the moment of production of the
mixtures, about 2 weeks before the feeding. The birds from
group 3 received mixtures with charcoal supplemented
during the feeding at a dose of 0.3% (together 100.3%). In
both cases the use of charcoal had a beneficial effect on the
development of the chickens. After 7 weeks of growth, birds
which received supplemented charcoal were from 22 g to
157 g (about 1 to 6.5 %) heavier, had a 5 to 9 % better feed
conversion ratio and a 1.6% better survival rate than the
control group. The Fattening Efficiency Index equaled 209
for the control group, 246 for group 2 and 262 for group 3.

Medycyna Weterynaryjna, 2003, vol: 59, number 1, pages

81-83
156 Evaluation of char and active carbon for the reduction of
ammonia volatilization from poultry manure. C. Ritz*, A. Tasistro,
B. Fairchild, and B. Bibens, University of Georgia, Athens.
Locally available biomass materials, such as peanut hulls or tree clippings,
have the potential for use as raw materials for producing chars
and active carbons. Chars have long been known to possess properties
that make them valuable environmental tools. One such application is
the adsorption of NH3. By properly activating the char, ammonia can be
captured on its surface. Active carbon can be found in numerous products
designed to absorb moisture and odors, such as carbon filters for use
in drinking water filtration and air purification. Chemical adsorption of
ammonia is mostly due to its interaction with oxygen functional groups
via hydrogen bonding. It is expected that chars used with poultry litter
application will perform mostly based on their chemisorption capacity.
The purpose of this study was to evaluate the effectiveness of char and
active carbon on reducing ammonia volatilization from poultry manure
when used as a surfaced-applied litter treatment. Char from peanut hull
waste was produced by pyrolysis, wherein the material was heated to
400°C in the absence of oxygen for 30 minutes. The unamended peanut
hull char (pH 9.20) and an acidified poultry litter char (53% sulfuric
acid) were applied to replicate pens at rates of 50, 75 and 150 lbs per
1000 square feet. Broilers were raised in the pens at 0.75 square feet per
bird to generate the manure ammonia. The unamended chars actually
increased the release of ammonia an average of 14% over the control.
The acidified char applied at the 50, 75 and 150 lb rates reduced overall
ammonia release by 11, 25, and 35%, respectively over the control. The
reduction in ammonia from the acidified char treatment is most likely due
to litter pH reduction from the acid application and not from chemisorption
by the carbon. Further investigation into the use of activated carbon
products for ammonia control in poultry houses is warranted.
Key Words: broiler, ammonia, char, active carbon, pyrolysis

Reference:
Poultry
Science
Vol
88
(Suppl
1)
,
p.
49.

2009
USDA ARS

Research Project: Agricultural by-Products As Adsorbents for

Environmental Remediation
Location: Commodity Utilization Research

Title: Efficacy of Activated Carbon from Broiler Litter in the

Removal of Litter Generated Ammonia

Submitted to: Meeting Proceedings

Publication Type: Proceedings/Symposium
Publication Acceptance Date: July 19, 2007
Publication Date: September 16, 2007
Citation: Fitzmorris, K.B., Miles, D.M., Lima, I.M. 2007. Efficacy of
Activated Carbon from Broiler Litter in the Removal of Litter
Generated Ammonia. In: Proceedings of the International
Symposium on Air Quality and Waste Management for Agriculture.
ASABE. 6 pages.

Interpretive Summary: Over the past 10 years, the production of

broilers has increased by 29 percent to approximately 9 billion in
2005. Ammonia (NH3) pollution from broiler excreta is a primary
concern for industry viability which requires innovative treatment
options. This research focused on the use of broiler litter as activated
carbon (BAC) to reduce aerial NH3 generated by litter, an
opportunity to not only reuse the manure, but also treat the
emissions from or within broiler houses. The objective of this study
was to evaluate the efficacy of BAC to remove NH3 volatilized from
litter samples in a laboratory acid-trap system. The BAC is a much
cheaper alternative than commercially produced activated carbons.
Preliminary studies using NH3/air mixture indicated that the BAC
NH3 uptake was approximately double that of Vapure 612, a
commercial carbon. In the litter emission study, the BAC and Vapure
performance was comparable. The NH3 emission reductions using the
activated carbon columns were 25% for BAC and 36% for Vapure
relative to the litter only control. The results of the study
demonstrate the potential for a cyclical waste utilization strategy in
using broiler litter activated carbon to capture NH3 volatilized from
litter.
Technical Abstract: Over the past 10 years, the production of broilers
has increased by 29 percent to approximately 9 billion in 2005. Ammonia
(NH3) pollution from broiler excreta is a primary concern for industry
viability which requires innovative treatment options. This research focused
on the use of broiler litter as activated carbon (BAC) to reduce aerial NH3
generated by litter, an opportunity to not only reuse the manure, but also
treat the emissions from or within broiler houses. The use of activated
carbon in the removal of NH3, specifically in broiler houses, has long been
discarded primarily due to the high cost and low efficiency of the carbons.
However, the study of BAC is a relatively new field that has focused on the
removal of organics and/or metals from water. The objective of this study
was to evaluate the efficacy of BAC to remove NH3 volatilized from litter
samples in a laboratory acid-trap system. The BAC is a much cheaper
alternative than commercially produced activated carbons. Preliminary
studies using NH3/air mixture indicated that the BAC capacity to adsorb
NH3 was approximately double that of Vapure 612, a commercial carbon.
In the litter emission study, the BAC and Vapure performance was
comparable. The NH3 emission reductions using the activated carbon
columns were 25% for BAC and 36% for Vapure relative to the litter only
control. The results of the study demonstrate the potential for a cyclical
waste utilization strategy in using broiler litter activated carbon to capture
NH3 volatilized from litter.



 Release of Inorganic Nitrogen and Phosphorus from
Poultry Litter Amended with Acidified Biochar
S.A. Doydora1, M.L. Cabrera1, K.C. Das2, J.W. Gaskin2, L.S. Sonon3, W. Miller1, and C. Steiner2
1Department of Crop and Soil Sciences, University of Georgia, Athens, GA 30602
2Department of Biological and Agricultural Engineering, University of Georgia, Athens, GA 30602
3Soil, Plant and Water Laboratory, 2400 College Station Road, Athens, GA 30602
sdoydora@uga.edu
Introduction
Georgia generates an estimated average of two million Mg poultry
litter (PL) annually. While this material is used as a fertilizer, its nitrogen
(N) value decreases as a result of ammonia (NH3) volatilization, which
may lead to losses of up to 60% of the applied N (Cabrera et al., 1993;
Cabrera and Chiang, 1994). One possible way of minimizing NH3
volatilization from PL is by mixing it with biochar (or char), a by-product
from bio-fuel production. When acidified, biochar may reduce NH3
volatilization from PL. This study aimed to 1) evaluate the effect of
acidified biochar on NH3 volatilization, 2) examine its effect on CO2
evolution and 3) determine its influence on the release of inorganic N
and P from surface-applied or incorporated PL.
Materials and Methods
Pine chip (PC) or peanut hull (PH) biochars (produced at 400 or
600°C) were acidified using 0.5 N HCl (1 g:10 mL). Poultry litter (2.1 g)
with or without acidified PC or PH (400 or 600) chars (2.1 g) were
surface-applied or incorporated into the soil (64.5 g) and was incubated
for 21 d. Volatilized NH3 was trapped in 0.1 N H2SO4 and measured
colorimetrically. Carbon dioxide evolved was measured using a CO2
analyzer. Release of inorganic N and P was determined by leaching the
soil with 0.01 M of CaCl2 followed by a N- and P-free nutrient solution
at 14 and 21 d of incubation. At the end of the incubation period, each
of the treatments was extracted with 1 M KCl.
Results
Surface-applied, unamended PL volatilized 17% of its total N while
PL amended with acidified biochars lost only 6 to 11% (Fig.1a). PC400
performed better than PC600 (F=20.92; p=0.0132) for reducing NH3
loss but there was no difference between PH400 and PH600 (F=2.12;
p=0.2293). When PL was incorporated into the soil, the same trends
were observed (Fig.1b).
Adding acidified chars to PL depressed CO2 evolved under surface
incubation (F=9.24; p=0.0083) (Fig.2a). Conversely, when char-
amended PL was incorporated into the soil, total CO2 was greater
compared to unamended PL (F=6.88; p=0.0255) (Fig. 2b).
Inorganic P in the leachate and final KCl extraction was not different
among treatments under both surface (F=0.68; p=0.6270) and
incorporated (F=1.08; p=0.4227) incubations (Fig. 3 and 4). However,
greater amounts of inorganic P were extracted from PL amended with
PC400 than from PL amended with PC600 for surface (F=43.66;
p=<0.0001) and incorporated (F=6.23; p=0.0341) incubations.
Inorganic N released from surface-applied PL with acidified chars
was comparable to that of unamended litter (F=3.05; p=0.1146) (Fig. 5).
PL+PC400 had more inorganic N leached than PL+PC600 (F=9.06;
p=0.0142). PL amended with PH biochar had greater inorganic N
leached compared to PL amended with PC biochar (F=6.74; p=0.0289).
No differences among treatments were observed under incorporated
incubation (F=2.56; p=0.1093) (Fig. 6).
Fig. 1. Cumulative NH3-N lost during a 21-d incubation
of poultry litter (PL, 2.1 g) with or without acidified chars
(PC400, PC600, PH400, PH600, 2.1 g) a) surface-
applied, or b) incorporated. (Error bars are standard
deviations.)
Fig. 3. Inorganic P leached from soil (s, 64.5 g) and
poultry litter (PL, 2.1 g) with or without acidified chars
(PC400, PC600, PH400, PH600, 2.1 g) a) surface-
applied, or b) incorporated. (Error bars are standard
deviations.)
Nitrate leaching was the same for all treatments under surface and
incorporated incubations (Fig. 5 and 6). Nitrate concentration in the leachate
was generally smaller than ammonium concentration particularly in the first
leaching. However, at the end of the studies, KCl-extracted concentrations
were larger for nitrate than for ammonium (Fig. 7).
Fig. 2. Cumulative CO2-C lost during a 7-d incubation
(20°C) of soil (s, 64.5 g) and poultry litter (PL, 2.1 g)
with or without acidified chars (PC400, PC600,
PH400, PH600, 2.1 g) a) surface-applied, or b)
incorporated. (Error bars are standard deviations.)
Fig. 4. Final inorganic P extracted from soil (s, 64.5
g) and poultry litter (PL, 2.1 g) with or without
acidified chars (PC400, PC600, PH400, PH600, 2.1
g) a) surface-applied, or b) incorporated. (Error bars
are standard deviations.)
Fig. 5. Inorganic N leached from Fig. 6. Inorganic N leached from Fig. 7 Inorganic N extracted from
soil (s, 64.5 g) and surface-applied soil (s, 64.5 g) and incorporated soil (s, 64.5 g) and poultry litter
poultry litter (PL, 2.1 g) with or poultry litter (PL, 2.1 g) with or (PL, 2.1 g) with or without acidified
without acidified chars (PC400, without acidified chars (PC400, chars (PC400, PC600, PH400,
PC600, PH400, PH600, 2.1 g) PC600, PH400, PH600, 2.1 g) PH600, 2.1 g) when a) surface-
during the a) first, or b) second during the a) first, or b) second applied, or b) incorporated. (Error
leaching. (Error bars are standard leaching. (Error bars are standard bars are standard deviations.)
deviations.) deviations.)
In the incorporated incubation (Fig 7b), char-amended PL had greater
amounts of inorganic N (mainly nitrate) extracted than unamended PL
(F=8.47; p=0.0173). Combining PL with PC600 and PH600 had larger
amounts of extracted nitrate compared to PL with PC400 (F=17.35;
p=0.0024) and PH400 (F=28.31; p=0.0005), respectively. However, in the
surface incubation, this was only observed between PH chars (F=13.29;
p=0.0054). Surface-applied char-amended PL did not increase the
amounts of inorganic N, or nitrate, extracted at the end of the incubation
(F=2.00; p=0.1908).
Conclusions
Amending PL with acidified chars:
  Reduced NH3 loss in PL by 63 to 36% with surface incubation and by 60
to 43% with incorporated incubation.
  Decreased CO2 evolved from surface-applied PL by as much as 21%
but increased that from incorporated PL by as much as 37%.
  Did not affect the release of both leachable and extractable inorganic P
from both surface-applied and incorporated PL.
  Did not affect the release of leachable inorganic N from PL under both
surface and incorporated incubations but led to greater amounts of
inorganic N, primarily in the form of nitrate, extracted from incorporated
PL.
References
Cabrera, M.L. and S.C. Chiang. 1994. Water content effect on
denitrification and ammonia volatilization in poultry litter. Soil Sci.
Soc. Am. J. 58:811-816.
Cabrera, M.L., S.C. Chiang, W.C. Merka, S.A. Thompson, and O.C.
Pancorbo. 1993. Nitrogen transformations in surface-applied poultry
litter: Effect of litter physical characteristics. Soil Sci. Soc. Am. J.
57:1519-1525.
Effects of feeding fowls with rice
hull charcoal on the egg-laying
performance and the odor of
chicken droppings.
Abstract; An investigation was conducted on the effects of feeding
laying hens with charcoal made from rice hull upon laying performance
and the odor of chicken droppings. When laying hens were fed with
feed containing rice hull charcoal at 1%, ammonia gas concentration in
accumulated chicken droppings and ammonia gas concentration in
chicken house were significantly lower than those of a control with no
addition of rice hull charcoal by 52% and 39%, respectively; and the
rate of hen-day egg production, daily egg production, and the fructure
strength of egg shell tended to be improved by 5%, 4%, and 8%,
respectively, compared with those of the control. On the other hand,
the amount of feed intake, body weight, Haugh Unit, egg shell weight,
and the amount of chicken droppings were not different from those of
the control, remaining within .+-.3% from those of the control. The
above result indicated that the addition of rice hull charcoal to feed at
1% was effective for the suppression of odor in accumulated chicken
droppings and within chicken house and for the improvement of the
rate of hen-day egg production and fructure strength of egg shell.

Accession number;03A0266617
Title;Effects of feeding fowls with rice hull charcoal on the egg-laying performance and
the odor of chicken droppings.
Author;SAITO KATSUMI(Aomori Prefect. Livest. Exp. Stn., JPN)
KUZUMAKI TAKEFUMI(Kitasato Univ.) HOSOKAWA YOSHIHARU(Kitasato Univ.)
Journal Title;Tohoku Agricultural Research
Journal Code:F0596B
ISSN:0388-6727
VOL.;NO.55;PAGE.141-142(2002)
Figure&Table&Reference;TBL.2, REF.3
Pub. Country;Japan
Language;Japanese
 Reasons for Pyrolysis of Poultry
Litter
• Traditionally, poultry litter is disposed by land
application and used as cattle feed
• Disposal of poultry litter in the U.S. poultry
industry is becoming a major problem because
of :
– Excess nutrient in the soil due to land application
– Contamination of drinking water
– Eutrophication of surface waters
– Ammonia emission from poultry houses
– Soil acidification through nitrification and leaching
– Biosecurity concerns
 Pyrolysis conditions
• Feedstock---air-dried poultry litter
• Temperature---450 to 500 oC
• Residence time--- 2 to 5 s
• Fluidizing medium--- nitrogen or producer
gas
• Feed rate --- 200 g/h
Bench-Scale pyrolysis reactor
system
Bio-oil derived from Broiler litter
Poultry litter biooil
Pyrolysis Char of Broiler Litter
 Bio-oil properties
Sample C H O N S (%) Moit pH Ash HHV
(%) (%) (%) (%) (%) (%) (MJ/kg

Chicken 55.25 6.54 37.58 <0.5 <0.05 5.3 2.7 <0.08 22.64
bedding

Flock-1 63.24 7.22 23.89 5.05 0.46 4.6 6.1 <0.09 28.25
litter

Flock-2 64.06 8.14 22.27 4.94 0.41 4.6 6.3 <0.09 28.0
litter

Broiler 62.84 8.31 20.72 7.23 <0.9 4.0 6.3 0.17 29.57
litter

Starter 64.90 8.44 20.31 5.60 0.4 3.7 4.2 0.10 29.76
turkey
litter
 2500

2000

1500
Viscosity (cP)

Neat Biooil
MeOH + Biooil
EtOH + Biooil
1000
Ace + Biooil

500

0
0 10 20 30 40 50 60

Time (days)
Potential Applications of bio-oil
 SYNTHESIS GAS OPTIONS
Cat: Ni, Fe, HYDROGEN
Cu-Zn
Cat: Cu-Zn, ETHANOL,
Bio-
Bio-oil Cu-Co MIXED ALCOHOLS

Cat: Cu-ZnO METHANOL, DME

Cat: H3PO4,
Cr2O3 OLEFINS
FEED PREP

LPG
Cat: Fe
NAPHTHA
GASIFICATION Cat: Ni SYNGAS FTL UPGRADING
KEROSINE/DIESEL
Cat: Co/K LUBES
WAXES
Cat: Cu-ZnO MeOH Cat: Zeolite GASOLINE
CLEANUP

Cat: Mixed Bases OXOCHEMICALS

Na, Ca
e.g., KETONES

CaCN AMMONIA

Cat: Ni/Mg SNG

Cat = Catalytic Combined

Cycle CHP
Conversion Process

Cofiring/Reburn CHP
 Biomass Pyrolysis (liquefaction)
Bio-Oil FromBio-oil Uses

Pyrolysis • Fuel oil substitute

• Chemicals
• Hydrogen
• Turbines (Power)

75 Green ton/day (40 Dry) Commercial RTP™

Facility at Rhinelander, WI operating since 1995 Oriented Strand Boards and Plywood
made from Bio-Oil – Phenolic Resins
are being tested at mill scale
Ensyn Technologies
 Flow chart of transportable pyrolysis unit
Exhaust to Atmosphere

Dolomite

Poultry Litter Hot Filtration Pyrolysis Oil

Fluidized Bed Cyclone Condensation
Poultry System
Pyrolysis Filter
Litter
Reactor System Cooli
Feeding
System ng
Tower
Ash
and
Char

Raw
Compress LBG LBG ESP and Oil Product
ed Reheat Compression Coalescing
Air Burner System

Excess LBG
to Feed
Startup
Dryer
LPG
Thermochemical conversion of biomass
to biofuels

Foster A. Agblevor,
Biological Systems Engineering
Virginia Tech, Blacksburg, VA.
Email: fagblevo@vt.edu

ICS-UNIDO Biofuels Workshop,

Accra, Ghana, Dec 11-13, 2007
 Outline of presentation
• Overview of pyrolysis phenomena
• Slow pyrolysis
• Fast pyrolysis of biomass
• Fast pyrolysis reactors
• Biomass pyrolysis products
• Applications of biomass pyrolysis products
 Outline of presentation
• Environmental application of pyrolysis
• Fractional catalytic pyrolysis
• Where do we go from here?
 What is pyrolysis?
• Pyrolysis is the thermal conversion of
organic materials in the absence of
oxidizing agents such as oxygen.
• Pyrolysis always occurs before any
combustion process
• Pyrolysis leads to thermochemical
decomposition of organic materials into a
complex mixture of compounds
Schematic depiction of Biomass
pyrolysis
 What is pyrolysis?
• Pyrolysis products are usually not well defined
especially when applied to complex natural
materials such as biomass.
• The pyrolysis phenomena can be divided into
several regimes depending on the heating rate
• Slow pyrolysis, fast pyrolysis, ultrafast pyrolysis,
vacuum pyrolysis and high pressure pyrolysis
 Fast pyrolysis biomass reactor
designs
• Bubbling fluid bed
• Circulating fluid bed
• Entrained flow
• Vortex
• Rotating cone
• Vacuum
 Biomass Feedstocks

Forest Wood Residues Agricultural Residues Energy Crops

Thinning Residues Corn stover Hybrid poplar

Wood chips Rice hulls Switchgrass
Urban Wood waste Sugarcane bagasse Willow
pallets Animal residues
crate discards
wood yard trimmings
Hybrid poplar nursery
 Biomass H3CO
H3CO

Constituents
HO OCH3 O
O O OH
OCH3OCH3 O
O HO OH
O
OCH3
OH OCH3
O O
HO
HO OH O OH
O O HO
HO O OH
O
OH OH OH
OH OH O
OCH3 H3CO HO
OCH3 O O HO

Lignin: 15-25% O
OCH3OCH3

OCH3
HO

O
HO
OH
O
OH

OH O
O
OH
HO
OH
O

OH O OH
O HO O HO
Complex aromatic structure O
O
OH OH
O
OH OH
O OH O
OH OH
O
HO OH OH O OH O

Very high energy content OH

H3CO HO
O
OH
O OH
HO
O HO
OH
O
OH
HO
O HO
OCH3 O O
OH OH
O OH O OH O
OH HO HO
O O HO O HO
O OH
O OH O OH
OH OH OH
O
OH
OH OH O OH O
HO HO
O O HO O HO
HO OH OH
OH
O O OH
O O
OH OH

Hemicellulose: 23-32%
O
HO OH O HO OH O
O O
O HO O HO
O OH O OH
OH OH
O OH OH
O
OH O OH O
HO HO

Polymer of 5 & 6 carbon OH

O
OH
O HO
OH
O
OH
O
OH
O HO

OH O HO OH O
sugar OH O
O HO
OH Solid
OH OH
O
OH O
HO
OH O HO
O
OH
OH O

Cellulose: 38-50% OH

Polymer of glucose, very good Gas or Liquid

biochemical feedstock
 Minor biomass constituents
• Extractives----2-10 wt%
• Ash----1-20 wt%
– N, P, K, Si, Ca, Cd, Hg, As
• These minor constituents strongly
influence the pyrolysis reactions
• quality of the pyrolysis oil
Environmental and fuel applications
Poultry Powered!!!
Broiler chicken litter
 Reasons for Pyrolysis of Poultry
Litter
• Traditionally, poultry litter is disposed by land
application and used as cattle feed
• Disposal of poultry litter in the U.S. poultry
industry is becoming a major problem because
of :
– Excess nutrient in the soil due to land application
– Contamination of drinking water
– Eutrophication of surface waters
– Ammonia emission from poultry houses
– Soil acidification through nitrification and leaching
– Biosecurity concerns
 Pyrolysis conditions
• Feedstock---air-dried poultry litter
• Temperature---450 to 500 oC
• Residence time--- 2 to 5 s
• Fluidizing medium--- nitrogen or producer
gas
• Feed rate --- 200 g/h
Bench-Scale pyrolysis reactor
system
Bio-oil derived from Broiler litter
Poultry litter biooil
Pyrolysis Char of Broiler Litter
 Bio-oil properties
Sample C H O N S (%) Moit pH Ash HHV
(%) (%) (%) (%) (%) (%) (MJ/kg

Chicken 55.25 6.54 37.58 <0.5 <0.05 5.3 2.7 <0.08 22.64
bedding

Flock-1 63.24 7.22 23.89 5.05 0.46 4.6 6.1 <0.09 28.25
litter

Flock-2 64.06 8.14 22.27 4.94 0.41 4.6 6.3 <0.09 28.0
litter

Broiler 62.84 8.31 20.72 7.23 <0.9 4.0 6.3 0.17 29.57
litter

Starter 64.90 8.44 20.31 5.60 0.4 3.7 4.2 0.10 29.76
turkey
litter
 2500

2000

1500
Viscosity (cP)

Neat Biooil
MeOH + Biooil
EtOH + Biooil
1000
Ace + Biooil

500

0
0 10 20 30 40 50 60

Time (days)
Potential Applications of bio-oil
 SYNTHESIS GAS OPTIONS
Cat: Ni, Fe, HYDROGEN
Cu-Zn
Cat: Cu-Zn, ETHANOL,
Bio-
Bio-oil Cu-Co MIXED ALCOHOLS

Cat: Cu-ZnO METHANOL, DME

Cat: H3PO4,
Cr2O3 OLEFINS
FEED PREP

LPG
Cat: Fe
NAPHTHA
GASIFICATION Cat: Ni SYNGAS FTL UPGRADING
KEROSINE/DIESEL
Cat: Co/K LUBES
WAXES
Cat: Cu-ZnO MeOH Cat: Zeolite GASOLINE
CLEANUP

Cat: Mixed Bases OXOCHEMICALS

Na, Ca
e.g., KETONES

CaCN AMMONIA

Cat: Ni/Mg SNG

Cat = Catalytic Combined

Cycle CHP
Conversion Process

Cofiring/Reburn CHP
 Biomass Pyrolysis (liquefaction)
Bio-Oil FromBio-oil Uses

Pyrolysis • Fuel oil substitute

• Chemicals
• Hydrogen
• Turbines (Power)

75 Green ton/day (40 Dry) Commercial RTP™

Facility at Rhinelander, WI operating since 1995 Oriented Strand Boards and Plywood
made from Bio-Oil – Phenolic Resins
are being tested at mill scale
Ensyn Technologies
 Flow chart of transportable pyrolysis unit
Exhaust to Atmosphere

Dolomite

Poultry Litter Hot Filtration Pyrolysis Oil

Fluidized Bed Cyclone Condensation
Poultry System
Pyrolysis Filter
Litter
Reactor System Cooli
Feeding
System ng
Tower
Ash
and
Char

Raw
Compress LBG LBG ESP and Oil Product
ed Reheat Compression Coalescing
Air Burner System

Excess LBG
to Feed
Startup
Dryer
LPG
Transportable Pyrolysis Unit
Rockingham County Cooperator
• A pilot pyrolysis unit will be demonstrated
on the property of our cooperator, Mr.
Oren Heatwole.
• The unit will be transportable from farm to
farm, but most of the initial research will be
done on-site.
• Poultry litter from a neighboring farm will
be used as the feedstock for the pilot
project.
Modified Furnace for Using Bio-
Oil
 ADI Stirling Engine System
Runs on Heat

Replace diesel Fuel Flexible

generators for
distributed power • Natural gas
market • Propane
• Ethyl alcohol
• Biomass
Quiet
• Hydrogen
• Waste heat
Output
shaft • Solar Heat
power to
Generator

Highest Efficiency of all heat engines with patented

“Dual Shell Pressure Balancing Technology”
 Fractional catalytic pyrolysis
• A catalyst is used for the fluidizing bed
• Catalyst fractionates biomass insitu
• Carbohydrates gasified to CO, H2, CH4,
CO2
• Lignin depolymerized and demethoxylated
to monomeric phenols
Schematic Diagram of Fluidized Bed Reactor

1. Fluidized Bed Reactor

2. Furnace

3. Thermocouple

4. Mass Flow Controller

5. Heat Exchanger

6. Hopper

7. Sample Feeder

8. Computer

9. Heating Tape

10. Cyclone

11. Reservoir

12. Condenser

13. Electrostatic Precipitator

14. AC Power Supply

15. Filter

16. Wet Gas Meter

17. Gas Chromatograph

Conventional wood pyrolysis oil C-13 NMR
Fractional catalytic pyrolysis oil
Lignin moieties in biomass
Molecular weight distribution of conventional and FCP oils

FCP oil

Fractionated sugar cane

Bagasse pyrolysis oil
 Acknowledgement
• We greatly appreciate the contribution of Virginia
Poultry Federation, Chesapeake Bay
Foundation and Shenandoah RC & Council for
their foresight in funding this project.
• National Fish and Wildlife Federation for Scale-
up funding support
• Mr Robert Clark for initiating the project,
collecting samples and getting the growers in
the Valley involved in the project.
• Waste Solutions Forum for promoting the project
 Acknowledgement
• Dr Sedat Beis
• Dr Seug-Soo Kim
• Dr Serpil Besler-Guran
• Ryan Tarrant
• Ofei Mante
• Frederick Teye
 Thank you
• Questions?
Fluidized Bed Reactor
 Viscosity of bioiol: Flock 1 litter, 450 ℃, 500 ℃,
550 ℃
450 500
550
16
50 oC 35
14 60 oC 50 oC
30 60 oC
16
12
Viscosity [ Pa.sec ]

50 oC

Viscosity [ Pa.sec ]
25 14 60 oC
10

20 12
8

Viscosity [ Pa.sec ]
10
6 15

8
4
10
6
2
5
4
0
0 100 200 300 400 500 0
0 20 40 60 80 100 120 2
-1
Shear rate [ min ]
Shear rate [ min-1 ] 0
0 100 200 300 400 500
-1
Shear rate [ min ]
 Viscosity of biooil ; Turkey Litter, 450 ℃, 500 ℃,
550 ℃

14
50 oC
60 oC
12

10
Viscosity [ Pa.s]

14
50 oC
8
12 60 oC

Viscosity [ Pa.sec ]
10

70
4
8 o
50 C
o
60 60 C
2
6

Viscosity [ Pa.sec ]
50
0
4
0 100 200 300 400 500
-1 40
Shear rate [ min ] 2
30
0
0 100 200 300 400 500
20
-1
Shear rate [ min ]
10

0
0 20 40 60 80 100 120
-1
Shear rate [ min ]
 Comparison of HHV of biooil and
litter samples
Raw sample Biooil HHV
Sample
HHV (Btu/lb) (Btu/lb)
Chicken 7,792 8,408
Bedding
Flock 1 6,514 11,829

Flock 2 7005 12,689

Broiler 6,781 12,040

Turkey 8,064 11,291

 Demonstration Unit
• We propose to use the funding from the National
Fish and Wildlife Federation to build a
transportable pyrolysis unit to convert poultry
litter into bio-oil and slow-release fertilizer in the
Shenandoah Valley.
• Work has began on the project.
• A demonstration unit will be built on the farm of
Mr Oren Heatwole, Poultry Specialties Inc,
Dayton, VA.
 Pyrolysis Project Overview
• The project seeks to demonstrate a
nutrient reduction and renewable energy
technology called pyrolysis to convert
poultry litter to a bio-oil, slow-release
fertilizer, and producer gas.
• This approach was identified as a priority
during the 2005 Waste Solutions Forum –
to create usable energy from animal
manure and poultry litter while also
reducing nutrients.
Site and Preparations for
Pyrolysis
Water and Heat Lines
 Slow pyrolysis

• The slow pyrolysis process is very old

and has been used since ancient times
to produce charcoal.
• In modern era, the slow pyrolysis
process has been modified and used to
produce charcoal.
• The modifications are focused on the
improvement in the yield of charcoal.
 Soil Biology & Biochemistry 36 (2004) 2067–2073
www.elsevier.com/locate/soilbio

Activated carbon amendments to soil alters nitrification rates

in Scots pine forests
L.M. Berglunda,*, T.H. DeLucaa,b, O. Zackrissona
a
Department of Forest Vegetation Ecology, Swedish University of Agricultural Sciences, 901 83 Umeå, Sweden
b
Department of Ecosystem and Conservation Sciences, The University of Montana, Missoula, MT 59812, USA
Received 20 February 2004; received in revised form 25 June 2004; accepted 28 June 2004

Abstract
The influence of charcoal on biotic processes in soils remains poorly understood. Charcoal is a natural product of wildfires that burned on a
historic return interval of w100 years in Scots pine (Pinus sylvestris L.) forests of northern Sweden. Fire suppression and changes in forest
stand management have resulted in a lack of charcoal production in these ecosystems. It is thought that charcoal may alter N mineralization
and nitrification rates, however, previous studies have not been conclusive. Replicated field studies were conducted at three late-succession
field sites in northern Sweden and supporting laboratory incubations were conducted using soil humus collected from these sites. We used
activated carbon (AC), as a surrogate for natural-occurring fire-produced charcoal. Two rates of AC (0 and 2000 kg haK1), and glycine (0 and
100 kg N as glycine haK1) were applied in factorial combination to field microplots in a randomized complete block pattern. Net nitrification,
N mineralization, and free phenol concentrations were measured using ionic and non-ionic resin capsules, respectively. These same
treatments and also two rates of birch leaf litter (0 and 1000 kg haK1) were applied in a laboratory incubation and soils from this incubation
were extracted with KCl and analyzed for NHC 4 and NO3 . Nitrification rates increased with AC amendments in laboratory incubations, but
K

this was not supported by field studies. Ammonification rates, as measured by NHC 4 accumulation on ionic resins, were increased
considerably by glycine applications, but some NHC 4 was apparently lost to surface sorption to the AC. Phenolic accumulation on non-ionic
resin capsules was significantly reduced by AC amendments. We conclude that charcoal exhibits important characteristics that affect
regulating steps in the transformation and cycling of N.
q 2004 Elsevier Ltd. All rights reserved.

Keywords: Activated carbon; Charcoal; Boreal forest; Birch litter; Fire; Nitrogen transformations; Nitrification; Phenolic compounds

1. Introduction cycling and changes in species composition, plant growth

There is limited understanding of the role of charcoal in
N mineralization and nitrification rates in boreal forest
ecosystems. Charcoal is a product of forest fires that
historically burned on a w100 years fire return interval in
northern Fennoscandia (Zackrisson, 1977) and amounts up
to 2000 kg haK1 of charcoal can be present in the soil of
such ecosystems (Zackrisson et al., 1996). Forest fires are a
major form of disturbance that has an immediate and long-
term effect on ecosystem processes. This effect vary
depending on fire severity, but involve alterations to nutrient
* Corresponding author. Tel.: C46-90-786-86-01; fax: C46-90-786-
81-66.
E-mail address: linda.berglund@svek.slu.se (L.M. Berglund).
0038-0717/$ - see front matter q 2004 Elsevier Ltd. All rights reserved.
doi:10.1016/j.soilbio.2004.06.005
and soil biota (Hart et al., 2004). The presence of fire in
these forests has been reduced considerably due to active
fire suppression and passive fire exclusion as a result of road
building and changes in forest stand management. A
prolonged absence of fire results in a change in understory
species composition shifting to a predominance of dwarf
ericaceous shrubs including Empetrum hermaphroditum
Hagerup, a ground cover of feathermosses such as
Pleurozium schreberi (Bird.) Mitt. and an increase in the
presence of Norway spruce (Picea abies Karst. (L.)) in the
overstory (Steijlen and Zackrisson, 1987; Linder et al.,
1997; DeLuca et al., 2002).
It is clear that fire temporarily increases N mineralization
and nitrification rates (Neary et al., 1999; DeLuca and
Zouhar, 2000; Choromanska and Deluca, 2001). The high
2068 L.M. Berglund et al. / Soil Biology & Biochemistry 36 (2004) 2067–2073
concentrations of available ammonium (NHC 4 ) present after
fire are produced chemically by soil heating during fire
and also by increased microbial activity following fire
(Choromanska and DeLuca, 2002). A pulse of available
nitrate (NOK 3 ) occurs within a year after fire and since NO3
K
is not produced directly by heating this is commonly
attributed to increased nitrification in excess of ammonium
(Wan et al., 2001). The long-term effect of fire on N
mineralization rates is not clear. Although fire appears to
reduce stocks of mineralizable N (Monleon et al., 1997;
Kaye and Hart, 1998; DeLuca and Zouhar, 2000) through
volatilization and fuel combustion, other changes in
N transformation are normally attributed to changes in
the composition of plant communities and soil biota (Hart
et al., 2004).
Many of the dwarf shrubs commonly found in late
successional Swedish boreal forests are known to produce
large quantities of phenolic compounds (Gallet and
Lebreton, 1995; Nilsson et al., 1998) that may directly
influence nutrient turnover and microbial activity (Wardle
et al., 1997). Polyphenolic compounds can affect N cycling
by stimulating microbial soil processes. Microorganisms
use polyphenols as a C source and with increased need of N
this promotes N immobilization. Formation of protein–
tannin complexes by polyphenolic compounds may also
impede mineralization and help to build up thick mor layers
of recalcitrant organic material (Northup et al., 1995).
The influence of charcoal on maintaining N availability
has been addressed by Wardle et al. (1998) and DeLuca
et al. (2002). Fire-produced charcoal has a porous structure
and sorptive surfaces which are shown to have the capacity
to adsorb significant amounts of polyphenolic compounds
(Zackrisson et al., 1996) and this effect is similar to
commercially-produced activated carbon (AC). Such
adsorption may result in deactivating the phenols through
chemical condensation of the phenols (Piccolo et al., 1999)
preventing toxic allelopatic effects and an increase in
microbial activity due to accumulation of dissolved C as a
food source and thereby increase mineralization. Charcoal
has been shown to harbour and support specific or unique
microbial communities (Pietikäinen et al., 2000), although
little is known on the functional differences of these special
communities. The porous structure of charcoal has also been
suggested to shelter microbial organisms from soil faunal
predators (Wardle et al., 1998).
Studies in our laboratory suggested that nitrification rates
are directly influenced by the addition of AC, used as a
surrogate for soil charcoal (DeLuca et al., 2002). In these
Table 1
General soil and site characteristics for each site
Site Location pH (GSE)
Reivo 65846 0 N, 19806 0 E 4.06 (0.03)
Vaksliden 65842 0 N, 18845 0 E 4.14 (0.02)
Strömforsheden 65808 0 N, 18853 0 E 4.06 (0.06)
short-term studies, we only observed a nitrification response
to AC at a relatively high and potentially unrealistic
application rate and the work was performed at only one
field site. Thus it is not clear if nitrification rates would be
significantly altered by AC additions to forest soils at lower,
naturally-occurring quantities. Furthermore, it was not clear
whether the presence of birch (Betula pubescence Ehrh)
litter (as is common in early succession) would also
influence nitrification rates as has been found in studies
involving aspen litter (Ste-Marie and Paré, 1999).
We hypothesized that charcoal would enhance rates of
nitrification in boreal forest soils. We used AC, as a
surrogate for natural-occurring fire-produced charcoal with
the intention of investigating the effect of AC on net N
mineralization and nitrification rates in Scots pine forests.
We also attempted to determine whether these effects were
related to AC adsorption of free phenolic compounds from
the soil.
2. Materials and methods
2.1. Site descriptions
Studies were conducted on a total of three forest stands in
northern Sweden. All sites are situated in forests reserves
within the northern boreal zone and consist of late-
successional Pinus sylvestris L. dominated forest of
ericaceous-cladina type on sandy glacial soils (Ahti et al.,
1968), classified as either Typic or Entic Haplocryods.
Pleurozium schreberi (Bird.) Mitt. and Cladina spp are the
dominant bottom-layer moss and lichen taxa and the field-
layer mainly consists of the ericaceous shrubs Empetrum
hermaphroditum Hagerup and Vaccinium vitis-idaea L.
Composite samples of the forest duff layer (Oe/Oa horizons)
were collected at each site and analyzed for pH, total C and
total N. General soil and site characteristics are presented in
Table 1.
2.2. Laboratory study
A laboratory incubation experiment was conducted to
assess the influence of charcoal and litter on net N
mineralization rates and nitrification rates in boreal soils
under controlled conditions. Samples from humus layer
were collected from the three P. sylvestris dominated sites.
The humus was sieved (!5 mm) to remove coarse roots,
mixed thoroughly and then air dried (20 8C) for 3 days.
Humus C (%) Humus N (%) Soil C (%) Soil N (%)
45.85 1.05 3.3 0.09
44.98 1.00 1.2 0.04
48.31 1.20 2.3 0.10
 L.M. Berglund et al. / Soil Biology & Biochemistry 36 (2004) 2067–2073
Then 10 g of humus (dry weight) was placed in 300 ml
(9 cm diameter) polyethylene jars. The humus was then
amended with one of the following treatments: 1) 0 or
100 kg N haK1 as glycine (Sigma Scientific) as an organic
N source; 2) 0 or 2000 kg AC haK1; 3) 0, or 1200 kg birch
litter haK1. This combination of treatments resulted in a
2!2!2 factorial experiment of 6 replicates for each of the
three sites, in total 144 jars. Treatments were applied by
mixing each component thoroughly with humus and then
placing the combined material into the polyethylene jar. The
AC was applied as 1.28 g of activated C (representing
2000 kg haK1) added to each pot and the glycine was
applied to the humus as a solution of 340 mg glycine
(535 kg glycine haK1 representing 100 kg N haK1) mixed
with distilled water. Dried birch leaf litter (Betula
pubescence Ehrh) was fragmented (4–5 mm) and 800 mg
was mixed into the humus. All the samples were adjusted to
a moisture content of 60% of maximum water holding
capacity (MWHC) and this was checked, and amended once
a week during the experiment. The jars were covered with
plastic lids with air-holes and incubated for 8 weeks at 13 8C
and 70% relative humidity. At the end of the experiment soil
samples were dried in 70 8C for 3 days and stored until
analysis.
2.3. Field study
Field experiments were carried out at the three late-
succession pine-dominated forests at forest reserves in
northern Sweden. Treatments consisted of two rates (0 or
100 kg N haK1) of glycine (Sigma Scientific) as an organic
N source and two rates (0 or 2000 kg AC haK1) of AC
resulting in a 2 by 2 factorial experiment. With ten
replicates of four treatments at three sites, this represent in
total 120 plots. Treatments were applied to 20 cm!20 cm
microplots from 0 to 10 cm depth within the forest floor as a
solution (glycine) or suspension (AC) by using a 50 ml
syringe with a 80 mm long, 2.10 mm wide steel needle. This
approach allowed for rapid incorporation of the AC
throughout the microplot with minimal disturbance to the
forest floor. The AC treatment was applied as 200 ml of the
40 g AC lK1 suspension. The glycine treatment was applied
as 100 ml of glycine solution (21.4 g glycine lK1). Distilled
water was added at the same rate as in the glycine and AC
treatment.
Net N mineralization and nitrification were monitored
in situ at these sites by using ionic resin capsules and free
polyphenols were monitored in situ by using non-ionic
carbonaceous resin capsules (DeLuca et al., 2002). This
approach allows NOK 3 , NH4
C
and polyphenols to be
assessed over a specific period rather than taking a single
point-in-time sample or using disruptive net N mineraliza-
tion techniques. One polyester capsules containing 10 ml
(approximately 1 g dwt) of mixed bed ionic resins (PST-2,
Unibest, Bozeman, MT) were placed at the bottom of the Oa
horizon or near the interface with the mineral soil in each
2069
microplot. Free phenols were monitored by placing one
polyester capsule (Unibest, Bozeman, MT) filled with
approximately 1 g dwt (about 1100 m2 of surface area) of
XAD-7 resin (Rohm and Hass Inc., Philadelphia, PA), near
the ionic resin capsule at the interface of the O horizon and
mineral soil in each microplot. In total, 120 ionic and 120
non-ionic resin capsules were used. In order to locate the
buried capsules, a thin nylon-line had been attached to each
resin capsule and the line was then attached to a marker
above ground when the capsule was buried. The resin
capsules were allowed to remain in the field for 2.5 months
from 11–18 June to 23 September, 2002. All capsules were
then removed from each plot, and stored at K20 8C until
analysis.
2.4. Chemical analyses
Forest O horizon samples were analyzed for total C and
N by dry combustion using an elemental analyzer
(PerkinElmer 2400 CHN, Norwalk, Connecticut, USA).
Soil pH was determined in distilled water using a 2:1
solution to soil ratio.
Soil mineral N in 2.5 g of the incubated soil samples was
extracted with 25 ml 2 M KCl on a shaker for 1 h, followed by
filtration through Whatman No 42 filters. The extractable
NHC 4 and NO3 were then analyzed by using a segmented flow
K
injection analyzer (Autoanalyzer III, Bran Luebbe, Chicago,
IL) and reported as mg NHC 4 –N or NO3 –N g soil
K K1
.
Ionic resin capsules were extracted for NH4 and NOK
C
3
analysis by placing each capsule into 10 ml of 1 M HCl and
shaking for 30 min. The extracting solution was then
decanted into a clean storage bottle and this process was
repeated for a second and third time to create a total
extraction volume solution of 30 ml. The extractant was
then analyzed for NHC 4 and NO3 as described above and
K
reported as mg NH4 –N or NO3 –N capsuleK1. Non-ionic
C K
resin capsules were analyzed for polyphenols following a
sequential extraction with water followed by 50% aqueous
methanol (Morse et al., 2000). Each capsule was placed in a
polypropylene centrifuge tube with 30 ml of distilled water,
shaken for 30 min, and the aqueous fraction was decanted
into a glass vial and stored at K20 8C until analysis. The
resin capsule was then immersed in 10 ml of 50% aqueous
methanol and shaken for 30 min. Extracts were decanted
into clean glass vials and this process was repeated for a
second and third time to create a total extraction volume of
30 ml. The methanol extracts were stored at K20 8C until
analysis for polyphenolic compounds. Polyphenol analysis
was performed by using the Prussian blue technique (Stern
et al., 1996) measured against catechin standards at 720 nm.
2.5. Statistical analysis
The N and polyphenol data were analyzed using
ANOVA. All data were analyzed for normality and
homogeneity of variance. Data found to violate assumptions
2070 L.M. Berglund et al. / Soil Biology & Biochemistry 36 (2004) 2067–2073

Table 2
Average pH in laboratory incubation from all three boreal forest sites,
incubated for 8 weeks

Treatment pH
0 3.62
CC 3.88
CL 3.85
CG 5.59*
CCCL 4.06
CLCG 5.62*
CCCG 5.75*
CCCGCL 5.73*

0Zcontrol; CCZAC addition; CGZglycine addition. Values followed

Fig. 1. Average soil NOK3 –N concentration (mg g
K1
soil) for all three boreal
forest sites in northern Sweden after 8 weeks of incubation. 0Zcontrol,
CCZAC addition, CGZglycine addition, CLZbirch litter addition.
Values followed by an asterisk are significantly different at P!0.05.
of ANOVA were analyzed using the non-parametric
Kruskal–Wallis test. Correlations were analyzed using
Pearson’s correlation test. All data analyses were
performed using SPSS Version 10.0 for Windows (SPSS
Inc. Chicago, IL).
by an asterisk are significantly different at P!0.05.
amendment of glycine and AC was 20% lower compared to
when only glycine were added (Fig. 2). Birch litter resulted
in the immobilization of added N (Fig. 2) and did not have a
significant effect on net nitrification (Fig. 1). Addition of AC
or birch litter did not significantly increase soil pH, but the
addition of glycine resulted in a slight, yet significant
increase in pH (Table 2).
3.2. Field study

3. Results Accumulation of NHC 4 on ionic resin capsules during the

2.5 months in the field was approximately 20 times higher in
3.1. Laboratory incubation plots, where glycine had been added as an organic N source
(Fig. 3). The glycine treatment also exhibited higher
Amendment of soils with AC in the presence of glycine amounts of NHC 4 –N (mg capsule
K1
) than did the treatment
increased net nitrification compared to the control and in the with both glycine and charcoal (Fig. 3). The NOK 3 –N

treatment, where glycine was added alone (Fig. 1). The accumulation on ionic resin capsules in the field was small
amount of NOK (Fig. 4) and there were no significant differences in the
3 in the presence of AC and glycine was
almost double (5–6 mg NOK compared to 3 mg amount of accumulated NOK between treatments or
3 –N g soil
K1 3

NO3 –N g soil ) that in the control treatment. Although

K K1 between sites. Total resin-sorbed polyphenols were signifi-
glycine greatly increased net ammonification (Fig. 2), this cantly lower on plots, where AC and glycine were added in
did not translate to an increase in nitrification unless AC was combination compared to the amounts in control plots
added. The amount of extractable NHC (Fig. 5). Amendment of glycine increased the amount of
4 –N in soil after
resin-sorbed phenols. Despite this increase, AC did adsorb
significant amounts of phenols. When AC was added alone

Fig. 2. Average soil NHC4 –N concentration (mg g

K1
soil) for all three boreal
forest sites in northern Sweden after 8 weeks of incubation. 0Zcontrol,
CCZAC addition, CGZglycine addition, CLZbirch litter addition.
Values followed by different letters are significantly different at P!0.001.
Fig. 3. Average resin-sorbed NHC 4 –N (mg capsule
K1
) for all three boreal
forest sites in northern Sweden after 2.5 month in the field. 0Zcontrol,
CCZAC addition, CGZglycine addition. Values followed by an asterisk
are significantly different at P!0.01.
 L.M. Berglund et al. / Soil Biology & Biochemistry 36 (2004) 2067–2073
Fig. 4. Average resin-sorbed NOK3 –N (mg capsule
K1
) for each of three
boreal forest sites in northern Sweden after 2.5 month in the field.
0Zcontrol, CCZAC addition, CGZglycine addition.
the amount of polyphenols was lower as well, although this
effect was not statistically significant (Fig. 5). There was no
clear correlation (data not shown) between net nitrification
rates and total phenols in the field.
4. Discussion
4.1. Laboratory incubation
Amendment of soils with AC in the presence of glycine
increased net nitrification compared to the control and in the
treatment, where glycine was added alone. Although
glycine greatly increased net ammonification this did not
result in an increase in nitrification unless AC was added.
This positive effect by AC confirms the findings by DeLuca
et al. (2002) and suggests that AC improves specific
mechanisms or environmental conditions that otherwise
limit the nitrification process and underscores the notion that
net nitrification in boreal forests is not limited by
availability of substrate (NHC 4 ). Nitrification in boreal
humus depends on a range of factors such as pH
Fig. 5. Average sorption of free polyphenols (mg capsuleK1) for all three
boreal forest sites in northern Sweden, after 2.5 month in the field.
0Zcontrol, CCZAC addition, CGZglycine addition. Values followed
by different letters are significantly different at P!0.05.
2071
and moisture, and is generally thought to be substrate-
limited. Many studies have shown a strong correlation
between the amount of NHC 4 in the soil and NO3 -
K
conversion (Currie, 1999; De Boer and Kowalchuk, 2001).
It is important to note that there was no increase in
nitrification with glycine alone in spite of the increase NHC 4
hydrolysis induced increase in soil pH following rapid
ammonification of the added glycine. Conversely, there was
no increase in pH with addition of AC alone, but there was a
significant increase in net nitrification when AC was added
with glycine. This suggests that pH by itself is not a major
determining factor for nitrification in our study, but may
reflect some synergy between the AC and increased pH
associated with the glycine. The positive effect of AC on
nitrification could also be a result of the adsorption of
phenolics, since phenolics have a generally negative effect
on nitrifying bacteria (Paavolainen et al., 1998). The main
negative effect of phenolics on nitrification is suggested to
be inhibition of enzymes crucial for transformation of NHC 4
to NOK 3 (White, 1994). It is also plausible that nitrifiers
aggregate around particles of AC, as AC can act as food
source supply for microorganisms due to the adsorption of
various soluble C substances. This aggregation of nitrifiers
could then produce a biofilm structure essential for
nitrification. De Boer et al. (1991), De Boer and Kowalchuk
(2001) have suggested that nitrifiers in acid environment
have developed a mechanism by which it is possible to
escape the negative effect of low soil pH on nitrification via
biofilm formation. Charcoal also have strongly hydrophobic
characteristics and could, when mixed into soil, alter
moisture conditions in soils, which may affect nitrifying
bacteria (Tate, 1995). Fierer and Schimel (2002) studied the
influence of repeated dry-wetting disturbance on nitrifying
bacteria and found an increase in population size with
increased number of disturbance events, and suggested that
nitrifiers are more drought-tolerant than other bacteria.
Our results show that the amount of extractable NHC 4 –N
in soil after amendment of glycine and AC was 20% lower
compared to when only glycine were added. Since
nitrification increased with addition of AC, a part of the
extractable NHC 4 is oxidized to NO3 . However, this does
K
not explain the whole difference, it is plausible that some of
the glycine could have been adsorbed by AC.
Birch litter resulted in an immobilization of added N.
This could be due to the high glucose content in newly shed
birch litter (Pietikäinen et al., 2000) which increased
microbial activity and subsequently increased the demand
for N. This did not occur in the treatment with no N
amendment, probably because here N was a limiting
resource. The addition of birch litter to the soil did not
have any significant effect on net nitrification. This result is
somewhat surprising because we had expected an increase
of nitrification after litter addition because birch stands
are often associated with higher nitrification potential.
Ste-Marie and Paré (1999) found that net nitrification rates
in deciduous forest floors were significantly higher than in
2072 L.M. Berglund et al. / Soil Biology & Biochemistry 36 (2004) 2067–2073
nitrifying coniferous samples. The reason for the lack of
response in our study could simply be due to the short time
period and the fact that we used coniferous humus samples
which could have buffered the nitrification response.
4.2. Field study
Ammonium accumulation on ionic resin capsules during
the 2.5 months in the field was approximately 20 times
higher in plots, where glycine had been added as an organic
N source. This was not surprising considering that boreal
humus is generally assumed to be N limited, and glycine is
an easily mineralized organic N source. The glycine
treatment also exhibited higher amount of mg NHC 4 –N
capsuleK1 than did the treatment with combined glycine and
AC amendment. Although this effect was not statistically
significant, it corresponds well to the results found in
the soil incubation study which showed a hampering effect
of AC.
The NOK 3 –N accumulation on ionic resin capsules in the
field was small and there were no significant differences in
the amount of accumulated NOK 3 between treatments or
between sites. This result diverges from the results found in
the laboratory incubation and from field investigations that
used higher rates of AC (DeLuca et al., 2002). Microbial
assimilation and plant uptake from areas outside these
microsites might have consumed excess of NOK 3 , and
masked any increase in NOK 3 production. Thus, there might
have been an increase in gross nitrification but not net
nitrification. Stark and Hart (1997) demonstrated that there
is large gross nitrification in late coniferous soil although the
net nitrification is hardly detectable. In our previous field
studie, a similar injection approach was used to distribute
the AC, however, the rate that effectively altered nitrifica-
tion was 5 times greater (DeLuca et al., 2002) than the rate
used in this study, thereby allowing for greater coverage of
the humus in the microplots.
AC addition did reduce the amount of total free
polyphenols in the soil. Total resin-sorbed polyphenols
were significantly lower on plots, where AC and glycine
were added in combination compared to the amounts in
control plots. Amendment of glycine increased the amount
of resin-sorbed phenols, probably because glycine may have
increased decomposition of polyphenolic compound and
thus increased the amount of free phenolics. Despite this
increase, AC did adsorb significant amounts of phenols.
However, the amount of polyphenols accumulated on the
resin capsules was highly variable and there was a site by
treatment interaction. There was no clear correlation
between net nitrification rates and total phenols in the
field as was found by DeLuca et al. (2002). Total free
phenols are a large and variable group of compounds, so it is
likely that only some of the phenolic compounds influence
nitrification. Further work would have to be performed to
determine how soluble phenolic compounds in boreal forest
soils actually influence nitrifier activity.
5. Conclusions
We conclude that charcoal exhibits important character-
istics that affect regulating steps in N cycling. In this study
we found that nitrification was limited both by a lack of
available NHC 4 and a physical or biochemical factor that
was altered by the AC amendment. Further studies are being
conducted to elucidate the mechanism responsible for
charcoal induced nitrification. Understanding how N is
regulated in fire-dominated ecosystems is important given
the dramatically altered fire regimes in northern Sweden, as
well as other portions of the globe, over the last century.
Acknowledgements
The authors wish to thank Derek MacKenzie and Tracie
Graafstra for their efforts in the laboratory.
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Biol Fertil Soils (2007) 43:303–311
DOI 10.1007/s00374-006-0106-5
ORIGINAL PAPER
Charcoal effects on soil solution chemistry and growth
of Koeleria macrantha in the ponderosa pine/Douglas-
fir ecosystem
Michael J. Gundale & Thomas H. DeLuca
Received: 26 July 2005 / Revised: 10 April 2006 / Accepted: 12 April 2006 / Published online: 3 June 2006
# Springer-Verlag 2006
Abstract We conducted laboratory and greenhouse experi-
ments to determine whether charcoal derived from the
ponderosa pine/Douglas-fir ecosystem may influence soil
solution chemistry and growth of Koeleria macrantha, a
perennial grass that thrives after fire. In our first experi-
ment, we incubated forest soils with a factorial combination
of Douglas-fir wood charcoal generated at 350°C and
extracts of Arctostaphylos uva-ursi with and without the
addition of glycine as a labile N source. These results
showed that charcoal increased N mineralization and
nitrification when glycine was added, but reduced N
mineralization and nitrification without the addition of
glycine. Charcoal significantly reduced the solution con-
centration of soluble phenols from litter extracts, but may
have contributed bioavailable C to the soil that resulted in
N immobilization in the no-glycine trial. In our second
experiment, we grew K. macrantha in soil amended with
charcoal made at 350°C from ponderosa pine and Douglas-
fir bark. Growth of K. macrantha was significantly
diminished by both of these charcoal types relative to the
control. In our third experiment, we grew K. macrantha in
soil amended with six concentrations (0, 0.5, 1, 2, 5, and
10%) of charcoal collected from a wildfire. The data
showed increasing growth of K. macrantha with charcoal
addition, suggesting some fundamental differences between
M. J. Gundale (*) : T. H. DeLuca
Department of Ecosystem and Conservation Sciences
University of Montana,
32 Campus Drive,
Missoula, MT 59812, USA
e-mail: mjgundal@yahoo.com
T. H. DeLuca
e-mail: tom.deluca@cfc.umt.edu
laboratory-generated charcoal and wildfire-produced char-
coal. Furthermore, they suggest a need for a better
understanding of how temperature and substrate influence
the chemical properties of charcoal.
Keywords Charcoal . Soil solution chemistry .
Douglas-fir and ponderosa pine ecosystems
Introduction
It is well-established that fire alters N cycling in the
ponderosa pine/Douglas-fir (Pinus ponderosa/Psuedotsuga
menziesii) ecosystem (Neary et al. 1999; Hart et al. 2005).
Nitrogen availability has been shown to increase immedi-
ately after fire (Covington and Sackett 1990, 1992; DeLuca
and Zouhar 2000) and may remain elevated on the scale of
months to years as a result of enhanced mineralization
(Covington and Sackett 1990, 1992; Monleon et al. 1997;
Kaye and Hart 1998; Gundale et al. 2005). Numerous
processes that increase N mineralization after fire have been
identified, including improved substrate quality (White
1991, 1994; Fernandez et al. 1997; Pietikainen et al.
2000a), death of roots and soil organisms resulting in a
large labile organic N pool (DeBano et al. 1979; Dunn et al.
1979; Diaz-Ravina et al. 1996; Neary et al. 1999), and a
reduction in C to N ratios due to preferential loss of C
during combustion (Gundale et al. 2005). A potentially
overlooked factor that may also enhance N cycling after fire
is the addition of charcoal to soils.
Several recent studies have shown that charcoal has the
potential to greatly enhance soil fertility. Amazonian forest
soils amended centuries ago with charcoal and manure still
maintain some of the highest biodiversity and productivity
304
of any soils within the Amazon basin (Glaser et al. 2001,
2002; Mann 2002). In boreal forest soils, charcoal was
shown to enhance N cycling by ameliorating the inhibitory
effects of litter extracts from late-successional species,
which in turn promotes growth of early-successional
species (Zackrisson et al. 1996; Wardle et al. 1998; DeLuca
et al. 2002; Berglund et al. 2004). Recently, DeLuca et al.
(2006) found that the addition of wildfire-formed charcoal
to ponderosa pine forest soils increased nitrification rates.
Charcoal may enhance soil fertility through a variety of
mechanisms. Increased N turnover may occur by charcoal
sorption of high C:N organic molecules from the soil
solution (Zackrisson et al. 1996; Wardle et al. 1998; Glaser
et al. 2002), resulting in reduced microbial N immobiliza-
tion and higher net mineralization and nitrification rates. In
addition, charcoal may remove specific groups of organic
molecules, including polyphenol or monoterpene com-
pounds that are thought to inhibit nitrification (Rice and
Pancholy 1972; Zackrisson et al. 1996; DeLuca et al. 2002;
Berglund et al. 2004). Sorption of organic molecules, along
with the gradual breakdown of charcoal, may initiate
humus formation and, thus, enhance long-term soil fertility
(Glaser et al. 2002). Charcoal may also enhance soil
fertility by creating habitat for microbes within its porous
structure (Pietikainen et al. 2000b).
Despite these potential roles that charcoal may have in
increasing soil fertility, its ecological role in forest ecosys-
tems, such as ponderosa pine/Douglas-fir, has received little
attention. We conducted three separate experiments using
low-temperature charcoal to investigate whether charcoal
influences soil solution chemistry and growth of an early
successional species. In our first experiment, our objective
was to determine whether charcoal had an influence on soil
solution chemistry after addition of the extracts of a late
successional species, Arctostapholos uvi-ursi, via surface
adsorption of phenolic compounds. We hypothesized that
charcoal added to a ponderosa pine forest soil will
effectively sorb the phenol fraction in litter extracts, which
would correspond with enhanced N cycling.
In our second experiment, our objective was to compare
the influence of charcoal made from the bark of two
species, ponderosa pine and Douglas-fir, on growth of
Koeleria macrantha, a perennial grass species that thrives
after fire disturbance in western Montana ponderosa pine/
Douglas-fir forests. Bark charring during low-intensity
wildfire is a potentially significant source of charcoal in
this system. Charred bark may gradually slough from trees
after fire and become incorporated in the soils surrounding
trees. It is recognized that ponderosa pine is a more fire-
adapted species than Douglas-fir; thus, an intriguing
hypothesis is that charred bark of the more fire-adapted
species will have a stronger positive effect on N cycling
processes and plant growth.
Biol Fertil Soils (2007) 43:303–311
In our third experiment, our objective was to determine
whether charcoal generated during a wildfire would have
any effect on K. macrantha growth and to determine
whether this relationship is dependent on soil charcoal
concentration. We hypothesized that wildfire charcoal will
positively influence K. macrantha and that this effect will
increase as a function of soil charcoal concentration.
Collectively, these three experiments address our central
hypothesis that charcoal will alter solution chemistry by
sorbing phenols and enhancing N cycling, which in turn
will improve the growth of early successional species.
Materials and methods
All three experiments utilized field-collected soil, which
was collected from the subsurface horizon (20–30 cm, Bw
Horizon) of a forest soil associated with low elevation
(1,100 m) ponderosa pine/Douglas-fir vegetation in western
Montana, USA. The soil is a sandy-skeletal, mixed, frigid
Typic Dystrustepts. This ecosystem is characterized by low
annual rainfall (<350 mm annually) with approximately
50% falling as snow during the winter months. Soil was
collected during the month of September, returned to the
lab, upon which they were sieved (4 mm) and homoge-
nized. We then added one part sand to three parts field
moist soil (by mass) to decrease fertility and increase
porosity and gas exchange, such that nitrification would not
be limited by low O2 availability. The sand fraction was
purchased as filter grade silica sand (for pool filters) and
was washed with 1 M HCl, followed by distilled water,
before being homogenized with field collected soil. This
sand-amended soil had a pH of 6.8, electrical conductance
of 91.2 μS m−1, and had a textural distribution of 71%
sand, 21% silt, and 8% clay.
All experiments also included the addition of either
laboratory-generated charcoal from Douglas-fir and ponder-
osa pine or charcoal collected in the field after a wildfire.
Laboratory charcoal was generated by burying wood or bark
from these species in silica sand and heating at 350°C for 2 h.
Charcoal was then ground and sieved as specified for each
experiment. Various physical and chemical properties of these
charcoals were measured (Table 1). Charcoal pH was
measured from a 4:1 slurry of deionized water to charcoal.
Electrical conductance (EC) was measured from charcoal
paste (2:1 distilled water and charcoal). Cation exchange
capacity (CEC) was estimated on charcoal samples via NHþ 4
replacement where 1 g of charcoal was rinsed twice with
25 ml of 1 M ammonium acetate (pH 7) to saturate exchange
sites. Excess saturating solution was removed from charcoal
samples with three consecutive washes with 25 ml of 95%
ethyl alcohol. Sorbed NHþ 4 was then extracted with 25 ml of
2 M KCl and analyzed on a segmented flow analyzer (Auto
Biol Fertil Soils (2007) 43:303–311 305

Table 1 Chemical properties of four charcoal types (df Douglas-fir, extract was 267.5 mg/l. Extract treatments received 25 ml
pp ponderosa pine, wildfire wildfire collected) used in the laboratory of this extract. No-extract treatments received an equivalent
and greenhouse experiments
volume of deionized water. Soils were homogenized
df Wood df Bark pp Bark Wildfire following this addition. This addition brought the soil in
each mason jar to a water content of approximately 60%
pH 4.15 4.18 4.81 5.74
WHC. Mason jars were incubated in the dark for 14 days
EC (μS g−1) 20.5 24.2 111.6 141.5
CEC (cmolc kg−1) 20.66 19.42 34.48 29.35
after which a portion of the soil was extracted and analyzed.
Density (g cm−3) 0.30 0.08 0.21 0.29 This entire experiment was repeated exactly as described
Total C (%) 72.9 72.4 71.5 82.3 above but with glycine added to all mason jars as a source of
Soluble phenols (μg g−1) 34.9 30.7 43.4 48.2 highly labile organic N to stimulate a more marked N
Total phenols (μg g−1) 441.7 148.1 810.4 393.8 response. Glycine, a simple amino acid that is readily
−1
PO3
4 (μg g ) 0.94 0.84 2.46 0.95 mineralized to NHþ
−1 4 , was added to each mason jar at a rate
NH4 (μg g )
þ
7.12 9.0 5.6 38.3
−1 of 75 mg/jar (250 mg/kg of soil). These two experiments will
NO3 (μg g ) 0.12 0.3 0.0 4.4
hereafter be referred to as the glycine and no-glycine trials.

Analyzer III, Bran Luebbe, Chicago, IL) using the Berthelot
reaction (Willis et al. 1993). Charcoal density was measured
by measuring dry mass of intact charcoal pieces and
measuring volume displacement in deionized water. Total
C was measured via dry combustion on a Fissions Elemental
Analyzer (Milano, Italy). Soluble and total phenols were
measured by extracting 1 g of ground charcoal in 25 ml of
deionized water and 50% methanol, respectively, and were
analyzed using the Prussian Blue Method (Stern et al. 1996).
Extractable NHþ 4 and NO3 (Mulvaney 1996) were extracted

by shaking 1 g of charcoal for 30 min in 25 ml of 2 M KCl,
and then filtering through Whatman #2 filters. The extracts
were analyzed for NHþ 4 –N using the Berthelot reaction
(Willis et al. 1993) and NO 3 –N by the cadmium reduction
method (Willis and Gentry 1987) on a segmented flow
analyzer (Auto Analyzer III). Soluble PO34 was extracted by
placing 1 g of charcoal in 25 ml of 0.01 M CaCl2 for 30 min.
Extracts were filtered through Whatman #42 filter paper and
then analyzed on a segmented flow analyzer using the
molybdate method as described by Kuo (1996).
Experiment 1: charcoal sorption potential
We conducted a laboratory incubation study using the soil
described above, where Douglas-fir charcoal and extract of
Arctostaphylos uva-ursi were added in a factorial combi-
nation yielding four treatments (Charcoal/Extract, Charcoal/
No extract, No Charcoal/Extract, and No Charcoal/No
extract). Each treatment was replicated five times and
consisted of 300 g of soil and placed into mason jars. The
treatments receiving charcoal addition received a 2%
charcoal amendment (20 g/kg). Charcoal was generated in
a muffle furnace by submerging Douglas-fir wood in sand
and heating it at 350°C for 2 h. Charcoal was ground and
sieved through a 4.75-mm sieve. A. uva-ursi extract was
made by extracting 100 g of A. uva-ursi leaves in 1 l of
deionized water for 24 h and filtering this extract through
Whatman #42 filters. The total phenol concentration of this
Experiment 2: effects of bark charcoal on plant growth
This greenhouse experiment consisted of three treatments
(Douglas-fir charcoal, ponderosa pine charcoal, and a
control) using the sand-amended soil described above to
evaluate the influence of charcoal source on K. macrantha.
Each treatment consisted of 20 replicate pots where each
pot received 1.5 kg of soil, and charcoal treatments received
a 2% (by mass) charcoal amendment. One percent of this
charcoal was homogenized into the soil, while the other 1%
was evenly distributed on the soil surface. We made
charcoal from Douglas-fir and ponderosa pine in the
laboratory by burying bark of each species in silica sand
and heating to 350°C in a muffle furnace for 2 h. Charcoal
was ground and sieved (<1 mm) using a Wiley mill.
Organic horizons (Oi, Oe, and Oa) were added to the surface
of each pot to add an additional and substantial mineraliz-
able pool of plant essential nutrients, as well as to provide a
source of bioavailable organic C that may influence soil
nutrient transformations. This organic material was ran-
domly collected (as described in Gundale et al. 2005) from
a ponderosa pine/Douglas-fir forest that had not been
exposed to fire for approximately 80 years and originated
from numerous species, including understory and overstory
species, but appeared to be primarily composed of
undecomposed ponderosa pine and Douglas-fir litter. The
organic material was homogenized and 100 g was added to
the surface of each pot. A mixed bed ionic resin capsule
(Unibest, Bozeman, MT) was placed in the center of each
pot to sorb nutrients throughout the duration of the
experiment.
K. macrantha was grown in these pots between October
2004 and March 2005 under ambient light conditions. An
average greenhouse temperature of 21°C was maintained. K.
macrantha seeds (Western Native Seeds, Coaldale, CO)
were germinated in a separate soil medium, and a single
seedling was transplanted into each pot. Pots were watered
3 days a week throughout the duration of the experiment. At
306
the end of the experiment, resin capsules were recovered,
and soil was rinsed from roots. Plants were oven-dried at 65°C,
and above- and belowground masses were measured.
Experiment 3: effect of wildfire charcoal on plant growth
Charcoal collected from a wildfire site was added to the soil
described above at a rate of 0, 0.5, 1, 2, 5, and 10%, and
placed in greenhouse pots seeded with K. macrantha to
determine whether an increase in soil charcoal content has
any influence on the growth of K. macrantha. Each
treatment (n=10) was established by adding 1.0 kg of
charcoal-amended soil per pot. The charcoal used in this
experiment differed from both previous experiments because
it was collected after a wildfire rather than generated in the
laboratory. Large particles (>5-cm diameter) of charcoal
were collected in the spring of 2004 from the Black
Mountain Fire (August 2003), Missoula, MT, (DeLuca et
al. 2006). It was impossible to decipher the species origin of
this charcoal, but it was likely primarily Douglas-fir and
ponderosa pine wood and bark char. The charcoal particles
were crushed, using a mallet, producing fragments ranging
from a diameter of 2 cm to microscopic. No attempt was
made to discriminate against any size class in an attempt to
simulate the range of charcoal particle sizes likely incorpo-
rated into the soil under natural conditions. Organic horizon
materials (50 g) were collected from a forest stand not
exposed to fire for over 80 years and added to the surface of
each pot as described earlier. All other experimental
conditions were run identically to experiment 2.
Laboratory analyses
At the end of experiment 1, 30 g of soil were extracted with
2 M KCl and analyzed for NHþ 4 and NO3 as described

above. Amino N was measured on these same extracts
using the ninhydrin method (Moore 1968). Soluble phenols
were extracted by shaking 30 g of soil for 1 h with 50 ml of
deionized water followed by filtration. Sorbed phenols were
extracted by shaking 30 g of soil with 50% methanol for
24 h followed by filtration. Phenols in these extracts were
measured using the Prussian blue method (Stern et al.
1996). Respiration was measured at the end of the
incubation by incubating 50 g dry weight equivalent soil
in a sealed container with 20 ml 1 M NaOH traps for 3 days
(Zibilske 1994).
Mixed bed ionic resin capsules (Unibest) were used in
experiments 2 and 3 to determine solution NHþ 4 ; NO3 , and

PO4 throughout the duration of the experiments. Capsules

were placed in the center of each pot, directly beneath each
plant, and were removed and extracted in 10 ml of 2-M KCl
three consecutive times. We analyzed NHþ 4 ; NO3 , and

PO4 from these extracts as described previously.
3
Biol Fertil Soils (2007) 43:303–311
Statistical analyses
Data in experiment 1 meeting assumptions of normality and
homoscedasticity were analyzed using two-factor analysis
of variance (ANOVA), where extract and charcoal were
entered as fixed factors under the general linear model.
Variables not meeting these assumptions were analyzed
using a Kruskal–Wallis test (K–W test). This analysis tests
for differences among treatments but does not evaluate the
significance of individual factors or interactions between
factors.
Data in experiments 2 and 3 were analyzed using one-
factor ANOVA followed by the Student–Newman–Keuls
post hoc procedure. Different letters are used to display post
hoc differences. Data not meeting assumption of normality
and homoscedasticity were compared using K–W tests,
which were not followed by post hoc procedures. All
analyses were conducted using SPSS 12.0 software.
Results and discussion
Experiment 1: low temperature charcoal sorption potential
Both charcoal and litter extract significantly influenced
numerous soil chemical variables (Fig. 1). In both glycine
and no-glycine trials, litter extract negatively influenced
extractable NO 3 concentrations. The negative influence of
A. uva-ursi on extractable NO 3 reported here is consistent
with our previous studies in ponderosa pine forest soils
(DeLuca et al. 2006) and with studies that showed that litter
from late-successional boreal species, such as the ericaceous
shrub Empetrum hermaphroditum, diminishes net nitrifica-
tion (DeLuca et al. 2002; Berglund et al. 2004). Charcoal
had an unexpected negative effect on NO 3 in the no-glycine
trial. In contrast, the addition of charcoal increased NO 3
concentrations in the glycine trial. These results may be a
function of the charcoal we used in this study, which was
generated at a low temperature (350°C). Charcoal contains a
significant concentration of bioavailable C, specifically
soluble phenols (Table 1) that may have caused net NO 3
immobilization (Schimel et al. 1996) in the no-glycine trial
where low NHþ 4 concentrations existed (Rice and Tiedje
1989). The NO3 immobilization effect did not occur in the

glycine trial because NHþ 4 limitations were drastically
reduced with glycine addition. In addition, higher rates of
nitrification in the glycine trial likely occurred because this
process was not limited by a lack of substrate availability
(glycine additions resulted in high NHþ 4 concentrations).
The higher rate of nitrification associated with charcoal
in the glycine trial is consistent with the finding reported by
DeLuca et al. (2006), which suggests that charcoal may
sorb compounds from litter extract and the soil solution that
Biol Fertil Soils (2007) 43:303–311 307

20 200
Amino N Amino N
a Amino N Charcoal *
b Charcoal ***

Extractable N (µg g-1)

Extract *
NH4+ Extract ***
15 Charcoal x Extract ** 150 Charcoal X Extract NS
NO3- Ammonium Ammonium
KW test *** Charcoal ***
Nitrate Extract **
10 Charcoal *** 100
Charcoal x Extract NS
Extract *** Nitrate
Charcoal x Extract * KW test ***

5 50

0 0

20 20
c Soluble Phenols
Soluble Phenols
d Soluble Phenols
KW test *
KW test **
Sorbed Phenols Sorbed Phenols Sorbed Phenols
Phenols (µg g-1)

15 15 Charcoal ***
Charcoal ***
Extract *** Extract **
Charcoal x Extract NS Charcoal x Extract NS

10 10

5 5

0 0

0.5 0.5
e f Respiration
Respiration (µg CO day )

Respiration
-1

Charcoal NS Charcoal NS
Extract * Extract NS
0.4 0.4
Charcoal x Extract NS Charcoal x Extract **
2

0.3 0.3

0.2 0.2

0.1 0.1

0.0 0.0
S S+E S+C S+C+E S S+E S+C S+C+E

No-Glycine Trial Glycine Trial

Fig. 1 Extractable amino N, NH4þ , and NO 3 [mean (SE)] without
(a) and with (b) glycine addition; soluble (water extracted) and sorbed
phenols (methanol extracted) [mean (SE)] without (c) and with (d) glycine
addition; and basal soil respiration [mean (SE)] without (e) and with
(f) glycine addition, from a 14-d soil incubation experiment where soils
were amended with a factorial combination of charcoal and extracts from
Arctostaphylos uva-ursi leaves (S soil only, S+E soil plus extract, S+C soil
plus charcoal, S+C+E soil plus charcoal plus extract). Data were analyzed
with a two-factor ANOVA where significance was tested for Charcoal,
Extract, and Charcoal × Extract interaction. Data that did not meet
parametric assumptions of normality or homoscedasticity were analyzed
using a Kruskal–Wallis (KW) test. Asterisks represent statistical signifi-
cance (p value, ns >0.1, *<0.05, **<0.01, ***<0.001)

are inhibitory to nitrifying bacteria, or sorb carbon-rich
molecules that would otherwise stimulate microbial immo-
bilization of N.
A. uva-ursi extract had a strong positive effect on NHþ4
in both no-glycine and glycine trials because it likely
contained some NHþ 4 and substrates that are rapidly
mineralized to NHþ 4 . Charcoal had a strong negative effect
on NH4 in both no-glycine and glycine trials. The
þ
mechanisms for this pattern may differ between the two
trials. In the no-glycine trial, the most likely explanation for
reduced NHþ 4 is that immobilization occurred as a function
of N limitations in these soils. In the glycine trial, higher
rates of nitrification associated with charcoal likely con-
tributed to lower NHþ 4 concentrations.
Both charcoal and extract significantly influenced con-
centrations of amino N that represent a highly labile
308 Biol Fertil Soils (2007) 43:303–311

Table 2 Plant mass and resin sorbed nutrients (mean±SE, n=20) from wood, such as lignin that are degraded during charcoal
a greenhouse experiment where soil was amended with 2% charcoal formation. It is unclear what effect these phenols have on
made from Douglas-fir (df) and ponderosa pine (pp) bark at 350°C
soil processes, but it is likely that they could be utilized as a
df pp no p food source by microbes, stimulating N immobilization.
Charcoal Charcoal Charcoal value Soil respiration showed little response to charcoal in
glycine or no-glycine trials. In the no-glycine trial, the extract
Total mass (g) a1.6 (0.2) a1.9 (0.1) b2.5 (1.0) <0.001
Root mass (g) a0.8 (0.1) a0.9 (0.1) b1.2 (0.1) <0.05
significantly increased soil respiration. Extract and charcoal
Aboveground a0.7 (0.1) b1.0 (0.1) c1.3 (0.1) <0.001 had no individual effect on soil respiration in the glycine trial;
mass (g) however, the interaction between charcoal and extract showed
Root to shoot a1.1 (0.1) b0.9 (0.1) b0.9 (0.1) <0.01 a significant effect. We speculate that this response may
ratio reflect that amines and degradable carbon substrates were
NHþ4 (μg resin a4.6 (0.9) b1.4 (0.6) b2.2 (0.8) <0.05 better utilized by microbes when phenolic molecules in the
capsule−1)
NO
3 (μg resin a1,770.8 b935.8 b581.5 <0.01 same extract were sorbed by charcoal.
capsule−1) (286.2) (241.9) (211.8) These data demonstrate that low-temperature charcoal
PO3
4 (μg resin a5.1 (1.4) a5.6 (0.9) b0.5 (1.2) <0.05 effectively sorbs soluble phenols from A. uva-ursi extracts,
capsule−1) which in turn stimulates nitrification, provided nitrification is
Letters in bold indicate differences using the Student–Newman–Keuls not substrate-limited. Our results are consistent with
post hoc procedure Berglund et al. 2004 and DeLuca et al. (2002), who showed
that the effect of charcoal on nitrification only occurred when
a labile N source was also present. These studies are also
consistent with the Terra Preta phenomenon reported in the
fraction of organic N that can be rapidly mineralized. Amazonian basin where charcoal and manure (high labile N
Glycine, which is a simple amino N molecule, stimulated concentration) were historically incorporated into the soil
rapid rates of N mineralization and resulted in increased (Glaser et al. 2001, 2002). Today, these soils maintain the
amino N concentrations, which suggests that the added highest fertility in the region, which may in part be a
glycine was not completely utilized and that substrate function of the interactive effect of charcoal and manure.
limitations were eliminated during this trial. In glycine and
no-glycine trials, the litter extract resulted in higher Experiment 2: effects of bark charcoal on plant growth
concentrations of amino N to soils. The effect of charcoal
on amino N, however, differed in glycine and no-glycine In this experiment, we unexpectedly found that charcoal
trials. In the no-glycine trial, charcoal significantly in- from both species diminished growth of K. macrantha
creased amino N concentrations. This response may have relative to the control with reduced mass in both above-
occurred because charcoal sorbed phenolic molecules that ground and belowground growth (Table 2). K. macrantha
otherwise would form insoluble complexes with amino N growing in pots with Douglas-fir charcoal had a signifi-
groups. In contrast, charcoal had a negative effect on amino cantly higher root to shoot ratio than the other treatments
N in the glycine trial, which is likely the result of charcoal that appeared to be primarily driven by low aboveground
enhancing microbial utilization of glycine. biomass. This data suggests that there is likely no difference
As expected, A. uva-ursi extract significantly increased in the effect of ponderosa pine and Douglas-fir charcoal on
phenols (soluble and sorbed) in both trials. The addition of plant species in this ecosystem.
charcoal to soil significantly diminished the soluble phenol We found that resin-sorbed NHþ 4 and NO3 were


concentration while increasing the pool of sorbed phenol.
This result is consistent with several studies in the boreal
forest that have demonstrated a high capacity of charcoal to
adsorb phenolic compounds (Zackrisson et al. 1996; Wardle
et al. 1998; DeLuca et al. 2002; Berglund et al. 2004).
Solubility of these fractions likely influences the degree to
which they are bioavailable and, therefore, their ability to
interfere with N transformations (Harborne 1997). It is
interesting to note that total phenols (sorbed and soluble)
was higher in the charcoal-only treatment of both trials than
the control, demonstrating that charcoal itself adds a
substantial amount of total phenol to the soil (Table 1).
These phenols are likely derived from the components of
significantly higher in the Douglas-fir charcoal treatment
relative to the ponderosa pine charcoal treatment and the
control. Resin-sorbed PO3 4 was significantly higher in both
Douglas-fir and ponderosa pine charcoal treatments than
the control. These results may be interpreted in several
ways. First, they may indicate higher mineralization and
nitrification rates in the presence of charcoal as suggested
by experiment 1. If higher mineralization occurred in the
presence of charcoal, it is unclear why a corresponding
increase in plant growth did not occur. It is possible that
some toxic substance was generated during charcoal
formation that inhibited root growth of K. macrantha,
despite a positive effect on nutrient availability (Fritze et al.
Biol Fertil Soils (2007) 43:303–311 309

Table 3 Plant mass and resin sorbed nutrients (mean±SE, n=10) from a greenhouse experiment where soil was amended with 0, 0.5, 1, 2, 5, and
10% charcoal collected from a wildfire
Percent charcoal

0% 0.5% 1% 2% 5% 10% p

Total mass (g) a0.5 (0.2) ab1.0 (0.3) ab1.1 (0.2) ab1.1 (0.2) b1.3 (0.1) b1.4 (0.1) <0.05
Root mass (g) 0.3 (0.2) 0.6 (0.2) 0.7 (0.1) 0.7 (0.1) 0.8 (0.1) 0.8 (0.1) >0.05
Aboveground mass (g) a0.2 (0.1) ab0.4 (0.1) ab0.4 (0.1) ab0.4 (0.1) b0.5 (0.1) b0.6 (0.1) <0.01
Root to shoot ratio 1.5 (0.2) 1.5 (0.3) 1.8 (0.2) 1.7 (0.3) 1.6 (0.2) 1.3 (0.1) >0.05
−1
NHþ4 (μg resin capsule ) 55.6 (4.0) 49.8 (4.6) 36.9 (6.0) 42.7 (2.2) 43.0 (1.3) 44.4 (2.9) <0.05a
NO3 (μg resin capsule−1)

a1,539.8 b947.9 bc552.3 bc556.1 bc561.8 c248.6 <0.001
(463.4) (128.4) (116.3) (93.4) (278.7) (29.8)
−1
PO3
4 (μg resin capsule ) a10.1 (1.4) a8.8 (2.1) ab5.8 (1.2) ab6.5 (1.7) bc1.7 (1.0) c0.0 (1.2) <0.001

Letters in bold indicate differences using the Student–Newman–Keuls post hoc procedure
All p values are for one-way ANOVA, unless otherwise noted
a
Kruskal–Wallis test p value

1998; Villar et al. 1998). These toxic substances are likely
to be more abundant in low temperature charcoals, such as
used in this experiment, and may be prone to volatilization
at higher temperatures. An additional explanation is that
charcoal may have enhanced soil macroporosity, allowing
more soil solution to pass through capsules, resulting in
misleading resin-sorbed nutrient concentrations.
Experiment 3: effect of wildfire charcoal on plant growth
In support of our hypothesis, natural charcoal collected from a
wildfire showed a positive effect on growth of K. macrantha
(Table 3). Both total and aboveground masses were signifi-
cantly higher in pots amended with 5 and 10% charcoal
addition than the control. Pots with lower charcoal content
(0.5–2%) showed an intermediate growth response. No
significant shift in allocation to above- or belowground
structures was detected across the charcoal gradient. As in
experiment 2, resin-sorbed NO 3 and PO4 decreased as plant
3
growth increased. These results suggest that these measure-
ments do not reflect any direct effect charcoal may have on
nutrient cycling, but are rather indicative of the solution nutrient
concentration as influenced by plant uptake. No difference in
resin sorbed NHþ 4 occurred across the charcoal gradient.
The different responses of K. macrantha to charcoal in
experiments 2 and 3 suggest that charcoal produced in a
laboratory may be greatly different from charcoal generated
during wildfire. Differences in charring conditions may
influence the chemical and structural nature of charcoal and
may therefore change its influence on soil solution
chemistry. One potentially important difference between
laboratory- and wildfire-collected charcoal was the ratio of
soluble phenols to NHþ 4 concentration extracted from the
charcoals (Table 1). While all charcoal had relatively
similar soluble phenol contents, which may stimulate
microbial N immobilization, high NHþ 4 concentrations
may have offset this immobilization effect when wildfire
charcoal was used. Another potentially important difference
is the different pH of laboratory charcoal and wildfire
charcoal (Table 1). The low pH associated with the lab
charcoals may have indirectly diminished P availability
in these treatments. Another difference between the
charcoal used in experiments 2 and 3 was the range
of charcoal particle size used. Experiment 3 incorporated
charcoal ranging from large (1–2 cm) to microscopic
fractions. We noted substantial root penetration into large
charcoal particles at the end of this greenhouse experiment,
which suggests that some resource, such as water, is more
available inside large charcoal particles. It is also possible
that grinding charcoal to a smaller size class, in some way,
eliminates its beneficial effects on soil fertility. For
instance, grinding may enhance the availability of organic
carbon because it is very immobile, whereas N ions are
significantly more mobile; thus, nutrient immobilization
may be more substantial when charcoal is ground.
Conclusion
It is clear that charcoal has the potential to significantly alter
soil solution chemistry and growth of K. macrantha. Charcoal
did not appear to stimulate N cycling in a low-nutrient
setting, but when glycine was added to soil, charcoal greatly
enhanced N mineralization and nitrification. This result may
indicate that low temperature charcoal contributes bioavail-
able carbon that causes N immobilization under low nutrient
conditions. As hypothesized, charcoal effectively sorbed
soluble phenols from litter extracts. This sorption may
effectively reduce the inhibitory effect of litter extracts on
soil microorganisms, plants, and biogeochemical processes.
Low-temperature, laboratory-generated charcoal had a nega-
tive effect on growth of K. macrantha, possibly as a result of
310
a toxicity effect caused by some compound formed during
low temperature charring or by N immobilization, as
suggested by the no-glycine soil incubation. In contrast,
charcoal created during a wildfire had a positive effect on the
growth of K. macrantha, suggesting low-temperature, labo-
ratory charcoal may not adequately represent field-collected
charcoal. Field-collected charcoal may have been generated
in a higher oxygen, higher temperature environment and may
have been exposed to leaching by rainwater and occlusion by
soil organic compounds before collection. Further investiga-
tion is required to evaluate how charcoal formation conditions
alter its effect on soil processes and plant growth and how
these processes manifest themselves in natural ecosystems.
Acknowledgements We thank V. Kurth, D. Mackenzie, and T.
Burgoyne for their assistance in the laboratory and greenhouse. We
also acknowledge funding from the NSF (NSF-DEB-03171108) and
the USDA Joint Fire Sciences Program (FFS #107) for this research.
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Springer 2005
DOI 10.1007/s11104-004-5485-5

Adsorption of allelopathic compounds by wood-derived charcoal:

the role of wood porosity

Olivier Keech1,2,4, Christopher Carcaillet3 & Marie-Charlotte Nilsson1

1
Department of Forest Vegetation Ecology, Swedish University of Agricultural Sciences, SE-901 83, Umeå,
Sweden. 2Present address: Department of Plant Physiology, Umeå Plant Science Centre, University of
Umeå, SE-901 87, Umeå, Sweden. 3Centre of Bio-Archaeology and Ecology (UMR 5059), CNRS/
Universite´ Montpellier 2, EPHE, Institut de Botanique, 163 rue Broussonet, F-34090 Montpellier, France.
4
Corresponding author*

Received 21 July 2004. Accepted in revised form 26 October 2004

Key words: adsorption, allelopathy, boreal forests, Empetrum hermaphroditum, germination, wood-
derived charcoal anatomy

Abstract
In Swedish boreal forests, areas dominated by the dwarf shrub Empetrum hermaphroditum Hagerup are
known for their poor regeneration of trees and one of the causes of this poor regeneration has been
attributed to allelopathy (i.e. chemical interferences) by E. hermaphroditum. Fire-produced charcoal is
suggested to play an important role in rejuvenating those ecosystems by adsorbing allelopathic com-
pounds, such as phenols, released by E. hermaphroditum. In this study, we firstly investigated whether
the adsorption capacity of charcoal of different plant species varies according to the wood anatomical
structures of these, and secondly we tried to relate the adsorption capacity to wood anatomical struc-
ture. Charcoal was produced from eight boreal and one temperate woody plant species and the adsorp-
tion capacity of charcoal was tested by bioassays technique. Seed germination was used as a
measurement of the ability of charcoal to adsorb allelochemicals. The charcoal porosity was estimated
and the pore size distribution was then calculated in order to relate the wood anatomical features to the
adsorption capacity. The results showed that the adsorption capacity of charcoal was significantly differ-
ent between plant species and that deciduous trees had a significantly higher adsorption capacity than
conifers and ericaceous species. The presence of macro-pores rather than a high porosity appears to be
the most important for the adsorption capacity. These results suggest that fire-produced charcoal has
different ability to adsorb phenols in boreal forest soil, and therefore may have differing effects on the
germination of seeds of establishing tree seedlings.

Introduction 1977). In Northern Sweden, early post-fire suc-

Wildfire is the most important disturbance factor
in boreal forests (Johnson, 1992; Payette, 1992)
occurring with 50–100 years of intervals in Fen-
noscandia (e.g., Engelmark, 1984, 1999; Hellberg
et al., 2004; Lehtonen and Huttunen, 1997;
Niklasson and Granström, 2000; Zackrisson,
* FAX No: +46-90-786-6676
E-mail: Olivier.Keech@plantphys.umu.se
cessions are usually dominated by Pinus sylvestris
L., Populus tremula L. and Betula pubescens
Ehrh. in the overstorey and, Vaccinium myrtillus
L. in the understorey. Long periods without fire
favour the growth of Picea abies (L.) H. Karst
and of the small fire sensitive dwarf shrub Empe-
trum hermaphroditum Hagerup (Haapasaari,
1988; Steijlen and Zackrisson, 1987). The fire
suppression over the last 100 years in Swedish
boreal forests has led to an intensive use of
292
mechanical soil scarification on clear-cuts and
has triggered the expansion and the dominance
of E. hermaphroditum (Zackrisson, 1977). E. her-
maphroditum is today one of the most common
species on northern inland clear-cuts (Data from
the Swedish National Forest Survey) where it
through chemical interference causes tree regener-
ation failures (Nilsson and Zackrisson, 1992).
Allelopathy, which involves the release of sec-
ondary plant metabolites, is a key process regu-
lating plant regeneration in many world
ecosystems (e.g., Fisher, 1987; Keeley et al.,
1985; Li and Romane, 1997; Pellissier, 1993;
Rice, 1979; Richardson and Williamson, 1988;
Waller, 1987; Wardle et al., 1998a). In the boreal
forests of northern Sweden, E. hermaphroditum
releases phenolic compounds, and in particularly
the dihydrostilbene Batatasin-III, from green
leaves and litter (Nilsson et al., 1998; Odén et al.,
1992; Wardle et al., 1998b; Zackrisson and Nils-
son, 1992) which inhibits seed germination and
seedling emergence, disturbs the plasmalemma
integrity of target roots’ cells (Wallstedt et al.,
2001) and negatively affects mycorrhizal symbio-
sis of coniferous trees (Nilsson et al., 1993). Fur-
ther, Batatasin-III impairs soil microbial activity
and slows down decomposition which both con-
tribute to an increased accumulation of soil
organic material with time since last fire distur-
bance (DeLuca et al., 2002; Zackrisson et al.,
1996). Many phenolic compounds also form
recalcitrant complexes with soil organic nitrogen
reducing the nitrogen accessibility to vascular
plants (Bending and Read, 1996; Gallet and
Lebreton, 1995; Wardle et al., 1998b). Therefore,
the phenol-regulated accumulation of humus and
the reduction of nitrogen available for plants
inhibit tree seedling establishment and growth of
P. sylvestris (Nilsson and Zackrisson 1992; Zac-
krisson et al., 1996), one of the most commonly
occurring tree species in the European and the
Asian boreal forests and one of the most impor-
tant species for the forest industry.
In field experiments, activated carbon has
been shown to adsorb phenols released by E. her-
maphroditum vegetation and to eliminate the
inhibitory effects of E. hermaphroditum on tree
seedlings establishment and growth (DeLuca
et al., 2002; Nilsson, 1994; Thoss et al., 2004;
Zackrisson and Nilsson 1992). When added to
field plots, activated carbon also increases humus
nitrogen mineralization and stimulates soil micro-
bial activity (Zackrisson et al., 1996). After a for-
est fire, up to 2000 kg ha)1 of wood charcoal is
produced (Zackrisson et al., 1996) and wood
charcoal produced by wild fire has similar prop-
erties as commercially manufactured activated
carbon (Bansal et al., 1988; Chereminisoff and
Ellerbusch, 1978). Fire-produced charcoal is thus
able to adsorb phenolic compounds released by
ericoid plants (Zackrisson et al., 1996). However,
the possible differential phenolic adsorption
capacities of charcoal produced by different
woody species have not been investigated before
whereas the wood anatomy varies between spe-
cies (e.g., Hellberg and Carcaillet, 2003; Schwe-
ingruber, 1990).
The aim of the present study is (1) to deter-
mine whether the charcoal from different plant
species differs in its ability to adsorb allelopathic
compounds produced by E. hermaphroditum and
(2) to investigate whether this adsorption can be
related to the wood charcoal anatomy of individ-
ual plant species. The ultimate aim of this study
is to contribute to a better understanding of the
functional role of fire-produced charcoal in the
boreal forest ecosystem.
Materials and methods
Plant species and production of charcoal
Twigs of eight plant species abundant in the
north European boreal forest were collected close
to Umeå, northern Sweden (6349¢N; 2018¢E).
The species consisted of five angiosperms (Betula
pubescens Ehrh., Empetrum hermaphroditum Hag-
erup, Ledum palustre L., Populus tremula L. and
Vaccinium myrtillus L.) and three gymnosperms
(Juniperus communis L., Picea abies (L.) H. Karst
and Pinus sylvestris L.). In addition, twigs of
Ulmus minor Mill. were collected in southern
France (4226¢N; 310¢E). Ulmus minor has been
selected for its very large vessels (Jacquiot et al.,
1973) and therefore provides a better possibility
to test the effects of vessels size on the adsorption
capacity of charcoal.
Twigs of 3–4 mm in diameter from each spe-
cies were cut into segments of approximately
30 mm of length and were then left to dry at
room temperature for 1 month. Charcoal was

produced in a ‘‘muffle furnace’’ (Nabertherm,
L9/S27) according to the following protocol:
wood fragments of each species were put in an
iron pan. Wood fragments from one species were
separated from the others by vertically inserting
a glass slide into the pan. The position of wood
samples was randomized in the pan and samples
were covered with sand to reduce the exposure to
oxygen during the burning process. This proce-
dure facilitates production of charcoal and
avoids total combustion of organic material and
production of ashes. The pan was put in the muf-
fle furnace for 35 min to reach 450 C and then
for an additional 15 min at 450 C after which
the charcoal samples were removed from the
muffle furnace. The selected temperature mimics
the temperature at the ground surface during
wildfire (Chandler et al., 1983; Wiedemann et al.,
1988). Then, charcoal samples were sieved to
retain material of 0.8–1.6 mm in size that corre-
sponds to the main size of soil charcoal (Carcail-
let and Talon, 2001). The burning procedure was
replicated three times for each species.
Adsorption of allelopathic compounds
The ability of charcoal to adsorb phenolic com-
pounds from an aqueous solution produced from
green leaves of E. hermaphroditum was deter-
mined by the use of a bioassay method following
Zackrisson and Nilsson (1992). In short, this
method involved collecting green leaves of E. her-
maphroditum (At Rovågern, N. Sweden, 6350¢N;
2015¢E), which were allowed to air-dry for 2
weeks. Deionized water was added to 50 g of dry
leaves per litre and the solution was agitated dur-
ing 48 h on a rotary shaker, and producing in
this way a 5% weight/volume water extract. This
solution was then filtered through a Munktell
No. 3 filter paper and diluted with deionized
water to produce a 2% solution E. hermaphrodi-
tum extract. This extract had a total inhibitory
effect on seed germination of Populus tremula
seeds (see below). For each species of charcoal,
0.4 g of charcoal fragments were added to 20 mL
of the 2% solution E. hermaphroditum and
placed on a rotary shaker for 12 h. Charcoal
fragments were then removed from the solution
by filtration through a filter paper (Munktell
No. 3) and 2 mL of the remaining solution was
added to each of five Petri dishes (50 mm in
293
diameter). This latter procedure is reiterated for
the three replicates of burning. To each dish,
25 P. tremula seeds (99.6% viability, stored
at )18 C) are added on a Munktell No. 3 filter
paper. The inhibitory effect of the solution on
P. tremula seeds was monitored and the number
of germinated seeds was used as a measure of
charcoal adsorption capacity (Zackrisson et al.,
1996). To verify whether the charcoal itself might
influence on P. tremula germination, five dishes
were set up with 2 mL of deionized water that
was soaked with 0.4 g of charcoal for 12 h; this
experiment is performed for all species. The Petri
dishes is placed in a climate chamber at 20 C
during 20 h per day of artificial illumination. The
total seed germination in each dish was recorded
after 7 days.
Charcoal porosity: estimation and size of pores
In this study, pores were defined as all longitudi-
nal cells that represent more than 95% of the
total wood composition in the selected species,
i.e. the vessels, the fibres and the parenchyma in
angiosperms and most of the tracheids in gymno-
sperms (Figure 1). The porosity of charcoal is
defined as a ratio between the total volume of all
pores and the total volume of wood. A transver-
sal section of each fragment of charcoal was used
to estimate the charcoal porosity. The total area
covered by pores was measured within an obser-
vation surface of 5250–5500 lm2. The ratio
between the total area of all pores and the obser-
vation surface is performed for each species of
charcoal to obtain a two-dimensional measure-
ment of porosity called ‘transversal porosity’.
This ratio serves to investigate whether the
adsorption capacity of charcoal is related to the
porosity of charcoal. A high value of this ratio
corresponds to a high charcoal porosity and con-
versely, a low value to a low charcoal porosity.
The charcoal porosity was measured on 10 trans-
versal sections for each species of charcoal.
The final ratio is thus based on the average of
the 10 measurements by species of charcoal. All
measurements were performed under an episcop-
ic-analysing microscope (magnification: ·500).
Values of surface were obtained with the image
analysing software ‘OPTIMAS 5.2’. The pore
area from each observation surface was used to
test whether the pores size might be related to
294

Figure 1. Scanning electronic microscope (SEM) pictures of transversal sections of wood charcoal. All pictures are at the same
scale and magnification (·200). The top three pictures (ericoids) show small diameter vessels and fibres. Betula pubescens and Popu-
lus tremula have larger but less abundant vessels. Elm (Ulmus minor) has little but very large vessels. The porosity of the three gym-
nosperms (Juniperus communis, Picea abies and Pinus sylvestris) is mostly composed of tracheids with thin wall in the early wood
and thick wall in the late wood.

the adsorption capacity. The pores are classified
according to the size of their area into micro-
pores (<50 lm2), meso-pores (from 50 to
250 lm2) and macro-pores (>250 lm2). This
classification is based on the frequency of pores
area per species (Figure 2). A ratio between
the total area occupied by each class of pores
and the transversal porosity was calculated.
The transversal porosity is thus defined as the
sum of three areas i.e. micro-, meso- and macro-
pores. Only charcoal produced from wood of
ericoı̈d plants (E. hermaphroditum, L. palustre
and V. myrtillus) and broad-leaved deciduous
trees (B. pubescens, P. tremula and U. minor)
were selected because conifers (gymnosperms) are
mostly composed of tracheids that do not show
such a distinction between micro-, meso- and
macro-pores.
Statistical analysis of data
Homogeneity of variances (Levene test) of data
was tested in accordance with the assumptions of
ANOVA. Firstly, data relative to adsorption
capacity of charcoal (Figure 3) was arsine square
root transformed prior to analysis. ANOVA were
performed to determine whether the adsorption
capacity of different species of charcoal was sig-
nificantly different from each other. Significant
differences between species (P
0.05) were log10
transformed and further analyzed by LSD test
(least significant difference). Secondly, ANOVA
used to determine whether the transversal poros-
ity of each species of charcoal was significantly
different between species of charcoal (Figure 4).
Significant differences (P
0.05) between species
are analyzed by Tukey test (honestly significant
difference). All statistics are computed with the
statistical package ‘SPSS 10.0’.
Results
The ability of charcoal to adsorb allelopathic com-
pounds in E. hermaphroditum leaf water extract
differs significantly (ANOVA: F8.18 ¼ 9.633;
 295

100

Populus
Betula
Ledum

Frequency of pores
80 Vaccinium
Empetrum
Ulmus

60
Meso-
pores Macro-
40 pores

20
Micro-
pores
0
0 50 100 150 200 250 300 350 400 500 1500 2500 3500

Transversal pore area (µm2)

Figure 2. Frequency of pores per area of wood charcoal of different plant species.

100.0
A
Adsorption capacity of charcoal (%)

90.0

80.0

70.0

60.0

50.0

40.0 BC
B
30.0
BC
20.0
CD D CD
D
10.0 D

0.0
m
s

ris
is
s
re
or

s
a

en

itu

lu

un

ie
ul

st
st
in

til
c

ab
em

od

e
lu

m
es
.m

yr

lv
pa

m
hr

P.
ub
tr

sy
m
U

co
ap
P.

L.
.p

V.

P.
J.
rm
B

he
E.

Figure 3. Adsorption capacity of charcoal from different plant species measured as the mean (±SE) number of germinated Populus
tremula seeds. The shaded bars correspond to angiosperm and open bars to gymnosperms. Data are expressed as the percentage of
seeds germinated in deionised water (control). Bars topped with different letters are significantly different from each other at P
0.05
(LSD test following ANOVA).

P
0.001) between species (Figure 3). With the highest adsorbing capacity amongst the tested
exception of Juniperus communis, the adsorption plant species, i.e. about three times higher than
of allelopathic compounds by charcoal produced P. tremula and about 15 times higher than P. syl-
from broad-leaved deciduous trees (B. pubescens, vestris. Juniperus communis adsorbs allelopathic
P. tremula and U. minor) are higher than the compounds in the same range as B. pubescens and
adsorption by charcoal from ericoid species P. tremula. The overall lowest adsorption capaci-
(E. hermaphroditum, L. palustre and V. myrtillus) ties are found for charcoal produced from V. myrt-
and conifers (P. abies and P. sylvestris) (Figure 3). illus and P. sylvestris (<5% of germinated
Charcoal produced from U. minor wood shows the P. tremula seeds). Adsorption capacity of charcoal
296

80.0
B
70.0 BD
BCD BCD

Transversal porosity (%)

ACD BCD
60.0 B AC
A
50.0

40.0

30.0

20.0

10.0

0.0
m

ns
s

is
ris
e

a
r

es
o

lu

tr

ul
itu

un
in

ce

st

bi
s
til

m
lu
od
.m

m
ve

a
es
yr

re
pa

P.
hr

yl
m

ub
U

co
P.

s
ap

L.
V.

.p

P.

J.
m

B
er
h
E.

Figure 4. Mean (±SE) of transversal porosity of charcoal from different plant species expressed as percent of the relative total area
of pores in the wood. Shaded bars correspond to angiosperms and open bars to gymnosperms. Bars topped with different letters
are significantly different from each other at P
0.05 (Tukey’s HSD test following ANOVA).

produced from J. communis, B. pubescens, P. abies representing more than 75% of the transversal
and E. hermaphroditum are not significantly differ- porosity (Figure 6). For E. hermaphroditum and
ent from each other at P
0.05 (Figure 3). V. myrtillus, species with relatively low adsorp-
The overall transversal porosity varies tion capacity, the surface of the sum of
between 47 and 67% among the species (Fig- meso-pores ranges between 40 and 50% of the
ure 4). The highest transversal porosity is found transversal porosity. The percentage of micro-
for P. abies and the lowest for P. sylvestris. The pores does not differ between species and ranges
values are significantly different from each other between 20 and 40% (Figure 6). Examples of
at P
0.05 (ANOVA: F8.81 ¼ 7.986; P
0.001). transversal wood pattern of selected species are
The transversal porosity for P. sylvestris is signif- displayed in Figure 1.
icantly lower than the transversal porosity
(<55%) of the other species (Figure 4). The
transversal porosity of U. minor is also relatively Discussion
low, but is only significantly different from
V. myrtillus, E. hermaphroditum and P. sylvestris. The results of the present study show that char-
When transversal porosity is tested against the coal of different plant species has differing capac-
adsorption capacity of charcoal, no correlation ity to adsorb phenolic compounds released by
between the two is evidenced (Figure 5). E. hermaphroditum. The adsorption capacity of
The frequency of pores per transversal pore charcoal from U. minor, deliberately selected for
areas displays three classes of pores that differ its large pores (vessels), is about double than all
between species (Figure 2). For the broad-leaved other species (Figure 3). Amongst the angio-
deciduous trees, the surface corresponding to the sperms, the adsorption capacity is higher for
sum of macro-pores represents more than 50% charcoal produced from broad-leaved deciduous
of the transversal porosity (i.e. U. minor and trees than charcoal from ericoid species. Most
P. tremula) and almost 40% for B. pubescens Ericaceous species are characterized by small
(Figure 6). The species with the largest relative pores diameter, whereas broad-leaved trees have
amount of macro-pores are also the species with larger pores diameter (Figure 1). The adsorption
highest adsorption capacity (Figure 3). Macro- capacity of gymnosperm charcoal differs signifi-
pores are almost non-existent in L. palustre, cantly among the selected species. We expected,
which is mainly constituted of meso-pores however wrongly, the total surface of adsorption
 297

90.0

80.0 U. m
Adsorption capacity of charcoal (%)
70.0

60.0

50.0

40.0 y = -0.7577x + 63.9

2
R = 0.0443
30.0
P. t
J. c
20.0
B. p

10.0 E. h P. a
L. p
P. s V .m

0.0
40 45 50 55 60 65 70

Transversal porosity (%)

Figure 5. Adsorption capacity and transversal porosity of charcoal from different plant species. U. m: Ulmus minor; P. t: Populus
tremula; B. p: Betula pubescens; E. h: Empetrum hermaphroditum; L. p: Ledum palustre; V. m: Vaccinium myrtillus; J. c: Juniperus
communis; P. a: Picea abies; P. s: Pinus sylvestris.

to be higher for those charcoal containing small macro-pores is generally higher for broad-leaved
diameter pores than those charcoal having large deciduous trees than for ericoid species where
diameter pores. This is because the relative macro-pores can be almost non-existent like in
porosity could have been lower when there was L. palustre (Figure 6). Charcoal species with a
a higher abundance of cell wall material. How- high density of macro-pores are also those with
ever, although different species differ in adsorp- the highest adsorption capacity, e.g., U. minor,
tion capacity (presumably due to wood anatom B. pub- escens and P. tremula (Figure 3 and 6).
ical differences), transversal porosity of the wood This observation suggests that a high adsorption
does not emerge as a driver of differences in capacity is linked with the total volume of macro-
adsorptive capacities among species because the pores. This result is strengthened by the observa-
process seems more complex and involves other tions of Chereminisoff and Ellerbusch (1978)
factors such as pore size distribution. If the total showing that adsorption capacities of charcoal
porosity was a significant factor explaining the were the results of both chemical and physical
adsorption, we should expect charcoal of P. properties. Macro-pores may have a lower surface
abies which had the largest transversal (Figure 4) tension than meso- and micro-pores, which should
to also have the largest adsorption capacity (Fig- in turn facilitate the penetration of water and dis-
ure 3), whereas this is not the case (Figure 5). solved compounds within charcoal.
Furthermore, the transversal porosity of P. abies Among the gymnosperms, J. communis and
and of P. tremula are not significantly different P. abies have a higher total porosity than P. syl-
from each other (Figure 4), while the adsorption vestris (Figure 4). SEM pictures (Figure 1) and
capacity is significantly different (Figure 3). literature on wood anatomy (Schweingruber,
The pore size distribution for each charcoal 1990) show that Pinus is generally characterized
species shows that the surface represented by by thick late wood relatively to the thickness of
298

100.0

90.0 Micro-pores
Meso-pores
Pore size distribution (%)
80.0
Macro-pores
70.0

60.0

50.0

40.0

30.0

20.0

10.0

0.0

m
ns

e
a
ro

lu

tr
ul

itu
in

ce

s
til
em

lu
.m

od
es

yr

pa
tr

hr

m
ub
U

P.

L.
ap

V.
.p

rm
B

he
E.

Figure 6. Pore size distribution in charcoal of six angiosperm species. Micro- ( lm2), meso- (50–250 lm2) and macro-porosity
(>250 lm2) are expressed as percentage of the total number of transversal pores of each species.

early wood, whereas Juniperus and Picea enced by other physical properties such as the
generally have a narrow late wood. Although the length of the pores (i.e. mostly length of vessels
diameter of conifer tracheids is relatively inde- for angiosperms and length of tracheids for gym-
pendent of the position within the tree ring, the nosperms), the density and the diameter of pit
wall thickness varies significantly which influ- orifices and the shape of the perforations that
ences the volume of the porosity. Our data indi- control the penetration of phenolic compounds
cate that species with a high proportion of late into charred cells.
wood material in their tree-rings appear to be To conclude, the present study supports previ-
linked to a lower adsorption capacity. ous work showing that fire-produced charcoal is
The differences in adsorption capacity able to regulate soil phenolic compounds released
between all species tested might also be influ- by E. hermaphroditum in the European boreal
enced by the presence of wood tar. Tar is forests (Pietikainen et al., 2000; Wardle et al.,
produced during the burning process and could 1998b; Zackrisson et al., 1996). Obviously, the
block the pores reducing the adsorption capac- simple volumetric process of adsorption occurs
ity of charcoal (www.fao.org/docrep/X5328f/ during the first days, but we stress the need to
x5328f00.htm#Contents) but also modify the understand the mechanism and kinetics of the
chemical properties of the charred surface. How- physico-chemical process of adsorption. The
ever, it is also likely that the different tree species identity of the plant producing the charcoal can
do not produce the same amount of tar during be important for determining the adsorption
the burning process, which also could explain capacity. Therefore, it is expected that variations
why the adsorption capacity is not linked to in seed germination and success of establishing
porosity. The presence of resins within conifers new trees seedlings are dependent on differences
trees might also influence the adsorption capacity in charcoal properties. It is likely that the regen-
of the charcoal produced, by modifying the inter- eration of trees is less important after a fire in
nal chemical structure of the pores. One of the areas dominated by ericoid species associated
limitations of the present study relates to the two with conifers than in areas dominated by broad-
dimensional estimation of the porosity that we leaved deciduous tree species notably those with
used. The adsorption by charcoal could be influ- large and abundant vessels, e.g., Ulmus, Quercus,

Fraxinus, Castanea in temperate forests and,
Salix, Populus, Betula and Sorbus in boreal
forests. The present study highlights the need for
conservation of broad-leaved deciduous trees in
boreal forests.
Acknowledgements
This research was funded through a European
Union scholarship to Olivier Keech and the
French Centre National de la Recherche Scientif-
ique (CNRS) to Christopher Carcaillet.
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Section editor: H. Lambers

Fire-Derived Charcoal Causes Loss of
Forest Humus
David A. Wardle,* Marie-Charlotte Nilsson, Olle Zackrisson
oreal forests serve as important global
BREVIA
Our results extend these findings by indicating
that charcoal can promote rapid loss of forest
humus and belowground C during the first dec-
ade after its formation. Fire often causes sub-
stantial losses of ecosystem C, and our results
provide evidence for a previously unreported mech-
anism that could contribute to these losses. Our
results are specific to boreal forests and to the type
of charcoal that we used, and further work is
needed to determine the importance of this mech-
versus Mix (exp)]. Further, nitrogen immobi-

B
sources or sinks of carbon (C), and wild-
fire is a major driver of C storage in these
forests. Although fire releases CO2 to the atmo-
sphere, it also converts plant biomass into forms
of black carbon, such as charcoal, that are resist-
ant to microbial attack and persist in the soil for
thousands of years (1). It has frequently been
lization was less than expected in the mixture
bags (Fig. 1). For these measurements, substrate
mixing effects [i.e., values for (observed – expected)/
expected] never differed significantly across the
three sites [P value always greater than 0.20
according to analysis of variance (ANOVA)]. This
result is despite the sites differing in both stand
anism in other biomes and for other types of
charcoal (11). Although several studies have
recognized the potential of black C for enhancing
ecosystem C sequestration (2, 3), our results
show that these effects can be partially offset by
its capacity to stimulate loss of native soil C, at
least for boreal forests. The effect of charcoal on
native soil C needs to be explicitly considered to

Downloaded from www.sciencemag.org on May 2, 2008

suggested that, because of its resistance, black C
can serve as an important long-term C sink that
may help offset the release of human-induced
CO2 to the atmosphere (2, 3). However, charcoal
is not biologically inert and can have important
effects on soil biological processes (4, 5). The
influence of charcoal on the decomposition of na-
tive soil organic matter remains poorly understood.
We conducted a simple experiment in each
of three contrasting boreal forest sites in northern
Sweden (6). Mesh bags were filled with pure
humus collected from the forest, pure charcoal
created in the laboratory, or a 50:50 mixture of
humus plus charcoal (6). These bags were left in
the field and harvested over 10 years. This ap-
proach is conceptually identical to that used for
the litter-mix studies that have greatly advanced
our understanding of the consequences of mix-
ing different litter types (7). This approach al-
lowscomparisons of observed valuesinthe mixture
with what would be expected on the basis of each
of the components of the mixture considered
separately.
We found that, over the 10-year period, loss
of mass and C from the bags containing mixtures
of charcoal and humus was substantially greater
than what was expected on the basis of the com-
ponents considered separately [Fig. 1, Mix (obs)
history and soil fertility (6) and points to similar
effects of charcoal across contrasting sites. Given
that charcoal decomposition rates in soil are ex-
tremely slow (2, 8) and that in our study system
charcoal persists for thousands of years in the
humus layer without evidence of mass loss (4),
most of the enhanced loss of mass and C caused
by mixing charcoal and humus must have resulted
from charcoal promoting humus loss rather than
humus promoting charcoal loss.
Substrate (i.e., glucose)-induced respiration
(SIR), a relative measure of active microbial bio-
mass (6), was always significantly greater in the
mixture bags than the value predicted on the ba-
sis of the charcoal and humus components con-
sidered separately [Fig. 1, Mix (obs) versus Mix
(exp)]. These results are consistent with charcoal
particles serving as foci for adsorption of organ-
ic compounds and microbial growth and activity
(4, 5), leading to enhanced decomposition rates
and mass loss of associated humus. The en-
hanced microbial activity in the mixture bags may
have led to greater mass and C loss through either
greater respiration or greater leaching of soluble
compounds (9).
Previous short-term laboratory studies have
shown that charred plant material causes accel-
erated breakdown of simple carbohydrates (10).
better understand the potential of black C as an
ecosystem C sink and agent of C sequestration.
References and Notes
1. J. W. Harden et al., Global Change Biol. 6 (suppl.), 174 (2000).
2. M. W. Schmidt, A. C. Nowak, Global Biogeochem. Cycles
14, 777 (2000).
3. J. Lehmann, Nature 447, 143 (2007).
4. O. Zackrisson, M.-C. Nilsson, D. A. Wardle, Oikos 77, 10
(1996).
5. J. Pietikäinen, O. Kikkila, H. Fritze, Oikos 89, 231 (2000).
6. Materials and methods are available on Science Online.
7. B. Gartner, Z. G. Cardon, Oikos 104, 230 (2004).
8. C. M. Preston, M. W. Schmidt, Biogeoscience 3, 397 (2006).
9. J. C. Neff, D. U. Hooper, Global Change Biol. 8, 872 (2002).
10. U. Hamer, B. Marschner, S. Brodowski, Org. Geochem.
35, 823 (2004).
11. K. Hammes et al., Global Biogeochem. Cycles 21, GB3016
(2007).
12. We thank A. Sundberg, K. Boot, G. Rattray, and
A. Mahomoud for technical assistance and T. Fukami,
M. Gundale, and anonymous reviewers for helpful
comments.
Supporting Online Material
www.sciencemag.org/cgi/content/full/320/5876/629/DC1
Materials and Methods
8 January 2008; accepted 3 March 2008
10.1126/science.1154960
Department of Forest Ecology and Management, Swedish
University of Agricultural Sciences, SE901-83 Umeå, Sweden.
*To whom correspondence should be addressed. E-mail:
david.wardle@svek.slu.se

Fig. 1. Changes in litterbag properties over a 10-year period. Humus, Charcoal,
and Mix (obs) correspond to litterbags containing pure humus, pure charcoal, and
a 50:50 mixture of charcoal and humus, respectively. Mix (exp) corresponds to
expected values for litterbags containing 50:50 mixtures of charcoal and humus if
no interactive effects between the components occur (6). Each data point is the
average of all three sites with 11 replicates per site, and vertical bars are mean
within-site standard error. For all measurements at all dates and sites, values for
Mix (exp) and Mix (obs) differ significantly at P = 0.01 (paired t tests). (A) Total
mass loss. (B) SIR. (C and D) Loss of C and N from litter bags (per unit initial mass)
over 10 years; negative values in (D) reflect net N gain through immobilization.

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