Journal of Environmental Management 111 (2012) 150e158

Contents lists available at SciVerse ScienceDirect

Journal of Environmental Management

journal homepage: www.elsevier.com/locate/jenvman

Pilot-scale passive bioreactors for the treatment of acid mine drainage: Efficiency
of mushroom compost vs. mixed substrates for metal removal
Hocheol Song a, Gil-Jae Yim b, Sang-Woo Ji b, Carmen Mihaela Neculita c, *, Taewoon Hwang c
a
Department of Environment and Energy, Sejong University, 98 Gunja-dong, Gwnagjin-gu, Seoul 143-747, South Korea
b
Geologic Environment Division, Korea Institute of Geoscience and Mineral Resources (KIGAM), 92 Gwahangno, Yuseong-gu, Daejeon 305-350, South Korea
c
Department of Civil and Environmental Engineering, Korea Advanced Institute of Science and Technology (KAIST), 335 Gwahangno, Yuseong-gu, Daejeon 305-701, South Korea

a r t i c l e i n f o a b s t r a c t

Article history: Pilot-scale field-testing of passive bioreactors was performed to evaluate the efficiency of a mixture of
Received 21 March 2012 four substrates (cow manure compost, mushroom compost, sawdust, and rice straw) relative to mush-
Received in revised form room compost alone, and of the effect of the Fe/Mn ratio, during the treatment of acid mine drainage
1 June 2012
(AMD) over a 174-day period. Three 141 L columns, filled with either mushroom compost or the four
Accepted 28 June 2012
Available online 11 August 2012
substrate mixture (in duplicate), were set-up and fed with AMD from a closed mine site, in South Korea,
using a 4-day hydraulic retention time. In the former bioreactor, effluent deterioration was observed over
1e2 months, despite the good efficiency predicted by the physicochemical characterization of mushroom
Keywords:
Pilot-scale passive bioreactors
compost. Steady state effluent quality was then noted for around 100 days before worsening in AMD
Acid mine drainage source water occurred in response to seasonal variations in precipitation. Such changes in AMD quality
Mixed substrates resulted in performance deterioration in all reactors followed by a slow recovery toward the end of
Mushroom compost testing. Both substrates (mushroom compost and mixtures) gave satisfactory performance in neutral-
Mn removal izing pH (6.1e7.8). Moreover, the system was able to consistently reduce sulfate from day 49, after the
initial leaching out from organic substrates. Metal removal efficiencies were on the order of Al
(w100%) > Fe (68e92%) > Mn (49e61%). Overall, the mixed substrates showed comparable performance
to mushroom compost, while yielding better effluent quality upon start-up. The results also indicated
mushroom compost could release significant amounts of Mn and sulfate during bioreactor operation.
Ó 2012 Elsevier Ltd. All rights reserved.

1. Introduction (Fe2O3), and magnetite (Fe3O4) at moderate pH (
3.5) accounts for

The use of passive bioreactors continues to increase worldwide
for the treatment of AMD (acid mine drainage)-impacted waters,
which are typically characterized by low pH and high metal and
sulfate concentrations. Passive bioreactors benefits include the ease
of installation and affordability, whereas the leading drawback is
their relatively limited lifetime (w10 years) (Thomas et al., 2010).
For typical bioreactors, the overall metal removal efficiencies
usually follow the order Al > Fe > Mn (Neculita et al., 2007), with
little dependence on the type of substrate materials. Precipitation
of Al as gibbsite [Al(OH)3] at pH > 5 and of Fe as oxi(hydroxi)des,
such as goethite (a-FeOOH), lepidocrocite (g-FeOOH), hematite
* Corresponding author. Present address: Research Institute e Mines and Envi-
ronment, University of Québec in Abitibi-Témiscamingue (UQAT), 445 Boul. de
l’Université, Rouyn-Noranda, Québec J9X 5E4, Canada. Tel.: þ1 819 762 0971x2278;
fax: þ1 819 797 4727.
E-mail addresses: hcsong@sejong.ac.kr (H. Song), gjyim@kigam.re.kr (G.-J. Yim),
swji@kigam.re.kr (S.-W. Ji), cmn@kaist.edu, Carmen-Mihaela.Neculita@uqat.ca
(C.M. Neculita), ht1029@kaist.ac.kr (T. Hwang).
most of their removal. Mn is generally acknowledged as hard to
treat due to various inhibitory factors, especially in the presence of
high concentrations of Fe (Edwards et al., 2009). These inhibitory
factors include the following: 1) the high solubility of Mn sulfide
(alabandite, MnS) (Stumm and Morgan, 1996); 2) the inhibition of
Mn precipitation under the condition of Fe/Mn >4 (Skousen et al.,
2000); 3) the anaerobic reductive dissolution of Mn oxides by
organics (Stone and Morgan, 1984), sulfides (Burdige and Nealson,
1986), and Fe(II) (Villinski et al., 2003); and 4) the relatively high
pH (>8) required for chemical precipitation as oxides (Mariner
et al., 2008).
The efficiency of AMD treatment in passive bioreactors relies on
sulfate-reducing bacteria (SRB), which require a source of available
organic carbon and appropriate environmental conditions (pH 5e8,
ORP <100 mV, and sulfate). Various waste materials (animal and
poultry manure, compost, woodchips and sawdust, and chitinous
materials) could be utilized as physical supports and substrates for
sustaining the growth of anaerobic bacteria, including SRB. Bacte-
rial activity improves mine water quality by increasing the pH and
reducing sulfate and metal concentrations. Among various waste

0301-4797/$ e see front matter Ó 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.jenvman.2012.06.043
 H. Song et al. / Journal of Environmental Management 111 (2012) 150e158 151

materials, spent mushroom compost, generally consisting of The pH was measured by mixing the solids with deionized water at
a mixture of rice straw, peat, animal manure (chicken, cow, or a 1:1 volume ratio (ASTM, 1995) and using a HACH pH-meter (HQ-
horse), gypsum, and limestone, is one of the most commonly used 40D). The water content of the organic substrates was measured by
organic substrates for anaerobic bacteria in the first generation of subtracting the weight of a sample dried at 105  C for 24 h from that
passive bioreactors (Dvorak et al., 1992; Stark and Williams, 1994; Ji before drying. The total volatile solids (TVS), which are usually used
et al., 2008). However, supplementation of easily available organic as an estimate of the organic matter content of solid materials, were
carbon is often required to compensate its depletion after contin- determined at 550  C as per Karam (1993). Dissolved organic
uous operation and, therefore, to extend the bioreactor lifespan carbon (DOC) was extracted from solid samples using deionized
(Stark et al., 1994). Recently, very high Mn removal efficiencies (up water and a solid: liquid ratio of 1:10 and a 2 h mixing period,
to 100%) were found in treating synthetic mine drainage (pH 6.2; following the procedure outlined by Zagury et al. (2006), and
90 mg L1 Mn) (Edwards et al., 2009; Karathanasis et al., 2010) and determined using an Apollo 9000 TOC analyzer (Teledyne Tekmar,
in synthetic AMD (pH 2.2; 567 mg L1 Mn) in the presence of Fe USA). Carbonenitrogenehydrogen content was measured with an
(Cheong et al., 2010). In addition, crab-shell chitin (high in elemental analyzer (Flash EA, Thermo Finnigan, USA). Before
carbonates and phosphates) was reported as a multifunctional elemental analysis, solid samples were prepared by drying and
(sorption media and substrate) amendment, offering significant grinding to a fine powder. The selected physicochemical properties
benefits to a mushroom compost-limestone-based bioreactor, of the organic substrates are presented in Table 1. Along with the
especially for Mn removal, when added in fractions of 50e100% substrates, creek sediment was used as a source of acclimated SRB,
(Robinson-Lora, 2009). However, highly variable Mn removal effi- which was collected from the bottom of a creek approximately
ciencies have been reported in the literature, ranging from 23% 1 km downstream from a coal mine located near Samchuk city
(Reisman et al., 2009) to 86% (Venot et al., 2008) in pilot-scale (South Korea). The sediment was kept in a headspace-free bottle
testing. Apparently, these inconsistent results are related to the and refrigerated at 4  C until use. Enumeration of SRB in organic
types of organic substrates used and to AMD quality and flow rate materials and the creek sediment was performed using the Most
(Guo et al., 2001; Neculita et al., 2011). Probable Number technique as per ASTM (2009).
There are approximately 300 coal mines and more than 2000
metal mines presently closed in South Korea. Mine drainage from 2.2. Pilot-scale bioreactor experiments
these sites has a wide range of pH values (2.1e7.4) and is generally
dominated by Fe (0.3e508 mg L1), Al (0.4e129 mg L1), and Mn Three pilot tests (Fig. 1) were performed over a 174-day period
(0e59 mg L1) (Ji et al., 2008; MIRECO, 2011). From 1996 to 2002, to evaluate the efficiency of a reactive mixture (in duplicate)
35 passive treatment systems at 29 mines were constructed to treat comprised of four organic substrates relative to mushroom
AMD, all of which utilized mushroom compost as a substrate compost alone (control), following the procedure detailed in
material (Ji et al., 2008). However, a relatively recent investigation Neculita et al. (2008) and Song et al. (2012). This reactive mixture
revealed highly variable removal efficiencies of acidity, sulfate, and was found to be highly efficient for pH neutralization and for
metals, depending on the substrate age, metal loading, and mine removal of sulfate and dissolved metals (Fe, Al, and Mn) during
drainage flow rates. Some systems also had various operational batch (35-day; Neculita et al., 2011) and laboratory (167-day)
problems such as overflow, leakage, clogging, broken pipes, and testing (Song et al., 2012). Unaltered AMD from a mine site in
inefficient sulfate and metal removal (Ji et al., 2008). Munkyoung County, South Korea was used as feed water for the
A comprehensive study was performed to identify alternatives bioreactors (Table 2). In order to provide a constant gravity-
to mushroom compost for AMD biological treatment in passive controlled flow rate to the bioreactors, the AMD was first stored
treatment systems (Neculita et al., 2011). Preliminary data obtained in a 200 L tank, from which the AMD flow rate was valve-controlled
in laboratory batch bioreactors indicated that a mixture of following weekly measurements. Columns (0.3 m in diameter and
substrates composed of cellulosic and organic wastes yielded 2 m in height) built with acrylic polymer with a total volume of
higher removal efficiencies for Mn and other metals relative to 141 L were initially packed with approximately 70 L of thoroughly-
mushroom compost (Neculita et al., 2011). This study is a follow-up mixed solid mixtures (1 m height for #1 and #2, and 1.2 m for #3),
investigation of the previous batch testing in an attempt to by slightly compacted layers of approximately 20 cm. The percent
demonstrate the enhanced performance of substrate mixtures in
treating a natural AMD source under scaled-up field conditions. The
testing was performed with pilot field-scale column bioreactors at Table 1
a closed mine site in South Korea over a 174-day period. The relative Characterization of natural organic materials used as substrates in pilot-scale
efficiency of a mixture of four substrates (cow manure compost, bioreactors.
mushroom compost, woodchips, and sawdust) compared to Mushroom Cow manure Cut rice Sawdust
mushroom compost for pH neutralization, sulfate reduction, and compost compost straw
metal removal was examined, with special attention to Mn removal Physicochemical parametersa
in both types of bioreactors. pH 7.64e7.67 6.64e6.70 6.11e6.16 4.78e4.85
Water content (%) 49.3 66.3 11.0 9.0
2. Materials and methods Total volatile solids (%) 55.8 79.2 84.6 99.2
DOC (mg L1) 3146 2011 2923 633
Elemental analysis (%)
2.1. Organic substrates and their characterization N 2.5 2.2 0.7 0.1
C 25.6 38.6 38.3 47.8
Cow manure compost, mushroom compost, sawdust, and rice H 3.3 5.6 5.7 6.6
S 1.8 < 0.01 < 0.01 < 0.01
straw were used as organic substrates. The composts (cow manure
C/N ratio 10 18 54 442
and mushroom) were obtained from a regional farmhouse located Microbiological counts (cells mL1)
in Hongseong city, South Korea. Sawdust and rice straw were Sulfate-reducing 1.7  103 7.0  101 <20 <20
purchased from Hanyoung Corp. (Seoul, South Korea). Rice straw bacteria (SRB)
pieces were provided uncut. Organic substrates were used without a
Physicochemical parameters were calculated as average values from two
pretreatment, while being kept refrigerated at 4  C until use. replicas.
152 H. Song et al. / Journal of Environmental Management 111 (2012) 150e158

Fig. 1. Schematic of downward-flow pilot-scale bioreactors.

composition of the mixture in reactors #1 and #2 was as follows: operation began with downward vertical flow with a 4-day
mushroom compost (20%), cow manure compost (10%), sawdust hydraulic retention time (HRT).
(20%), rice straw (10%), creek sediment (20%), and limestone (20%),
whereas reactor #3 received 100% mushroom compost. Note that 2.3. Sampling and analyses
the mass of organic substrates was corrected for water content
when mixed with other solids. To prevent clogging at the effluent The first sample was collected at the end of the first week, i.e.
port, the lower ends of the columns were filled with coarse quartz after the acclimation period. Samples were then collected weekly
gravel (2e5 cm diameter), which consists almost entirely of quartz, (for a total of 20 sampling-events) from the AMD reservoir and
to a height of around 30 cm. Also, an 80-mesh (0.177 mm) nylon from the exit end of the effluent tubing, which drained to an
disc was placed on the bottom of the gravel layer to prevent loss of effluent pond. Unfiltered samples were immediately measured for
organic particles to the effluent port. The columns were then pH (Hach PHC101-01, USA), redox potential (Hach MTC101, USA),
saturated with the mine-site AMD, by allowing the water to flow dissolved oxygen (DO) (LDOTM), electrical conductivity (CDC401),
vertically downward through the solid material, while slightly total dissolved solids (TDS) (CDC401), and salinity (CDC401) or
inclined or gently shaken to remove air pockets within the were preserved for culturable SRB counts (at every two sampling-
substrate layer. The AMD feed was then stopped for one week for events). The recorded values of oxidation-reduction potential
bacterial acclimation. This acclimation period was necessary to (ORP, mV) were converted into Eh by adding the reference elec-
grow a large population of SRB that would produce enough sulfide trode potential (Eref) of 210 mV, corresponding to a 3 M KCl solu-
and alkalinity in order to withstand the shock of the AMD feed tion. A portion of the samples (about 10 mL) was filtered with
(Waybrant et al., 2002; Neculita et al., 2008). After acclimation, the 0.45 mm membrane mixed cellulose ester filters (Whatman, USA).
The filtrate was then immediately analyzed for sulfide (methylene
blue method), sulfate (sulfaVer 4 method), and ferrous iron (ortho-
Table 2 phenanthroline method) or was preserved for DOC, metal (Al, Fe,
Composition of acid mine drainage (AMD) fed into pilot-scale bioreactors.
and Mn), and micronutrient (Na, K, Ca, and Mg) analysis in the
Component Units Valuesa laboratory. Detection limits of the methods were as follows:
pH e 2.93e4.07 5 mg L1, 1 mg L1, and 0.02 mg L1 for sulfides, sulfate, and ferrous
ORP mV 461  41 iron, respectively. Metal concentrations were measured using
DO mg L1 6.4  2.0 inductively coupled plasma (HORIBA Jobin Yvon, France) on an
Conductivity mS cm1 2.1  0.4
TDS mg L1 1118  174
acidified sample with concentrated HCl (1:9). Unless stated differ-
Salinity & 1.1  0.2 ently, all parameters were determined using standard methods
Fe2þ mg L1 114.2  58.8 (APHA, 2005). Culturable SRB enumeration was performed with an
Al3þ mg L1 97.2  34.9 unfiltered sample using the Most Probable Number (MPN) tech-
Mn2þ mg L1 29.0  8.2
nique as per ASTM (2009). A modified Postgate C was used as
Ca2þ mg L1 310.7  94.2
Mg2þ mg L1 170.0  49.7 growth media, with a pH 7 and containing lactate as carbon source.
Naþ mg L1 91.7  146.5 More details on its composition are provided in Neculita et al.
Kþ mg L1 9.4  4.4 (2011). It should be noted that the MPN counts in the liquid
SO2
4 mg L1 1277  726 samples might give only limited information about the bacteria in
a
Mean and standard deviations were calculated with n ¼ 20. the reactor since the majority of the SRB probably lived in a biofilm.
 H. Song et al. / Journal of Environmental Management 111 (2012) 150e158
MPN counting in the effluent gives information about detachment
rates rather than active bacteria. All reagents were of ACS quality.
3. Results and discussion
3.1. Characterization of natural organic materials
Consistent with previously reported findings, the sawdust had
a typical wood waste acidic pH (4.8) (Table 1), while other organic
waste compost (mushroom and cow manure) had neutral pH
values (6.6e7.7) (Neculita and Zagury, 2008). On the other hand,
the pH of cut rice straw (pH 6.1) fell between the two groups of
natural organic materials. Based on the TVS values, the sawdust
consisted of more than 99% organic material and cut rice straw
consisted of 85%, whereas the mushroom compost had only 56%,
with the leftovers of this last substrate consisting of inorganic
materials such as calcite and gypsum, in additions to ashes. Yet, the
DOC concentrations (the higher DOC, the more organic carbon is
available for microbial activity) released in water from mushroom
compost (and eventually available for bacterial consumption) were
the highest (3 g L1), which is within the typical range of
0.8e11 g L1 (Guo et al., 2001). This observation, together with a C/
N ratio of 10 (the optimal value for the biodegradation of complex
organic materials), indicated that the mushroom compost was the
best substrate (Cheong et al., 2010), followed by cow manure
compost. However, in a recent study, mushroom compost showed
poor performance in sulfate removal and acid neutralization of
mine drainage (Robinson-Lora, 2009). Furthermore, mushroom
compost was proven to be a good substrate for AMD in the short
term but a poor one over the long-term operation of passive
bioreactors (Ji et al., 2008). Similarly, the poor performance of
manure as a sole substrate has also been reported, despite its high
DOC concentrations and the favorable C/N ratio (Zagury et al.,
2006). In fact, in field passive systems, a higher initial DOC
Fig. 2. Effluent chemistry changes during AMD treatment in pilot-scale bioreactors. Reactors #1 and #2: mushroom compost (20%), cow manure compost (10%), sawdust (20%), rice
straw (10%), creek sediment (20%), and limestone (20%); Reactor #3: mushroom compost (100%).
153
concentration could entail a faster depletion of the source of
available organic carbon for anaerobic bacteria. Bacterial counts
showed that culturable SRB were present only in compost (cow
manure and mushroom; up to 2  103 cells mL1), which is
consistent with previously reported results (102 cells mL1 in
composted poultry manure and leaf compost) (Neculita and Zagury,
2008).
3.2. Pilot-scale bioreactor tests
3.2.1. pH, Eh, DO, conductivity, TDS, and salinity
pH values sharply increased from <4 to a moderate range and
remained in the moderate region (6.1e7.8) despite temporal vari-
ation throughout the entire testing period. The pH profile of the
reactors generally followed the pH changes of the source AMD,
with the exception of reactor #2, which registered an unusually low
pH (4.6) on day 152 (Fig. 2). An increase in pH is typically observed
in bioreactors (Waybrant et al., 2002; Neculita et al., 2008; Song
et al., 2012). This increase is associated with the alkalinity
increase that results from the formation of carbonate species either
through dissolution of limestone from reactive mixtures or through
the oxidation of readily available organic substrates by SRB, as well
as with the dissolution of the surface-bound hydroxyl ion from
substrate materials, under acidic conditions (Neculita et al., 2011).
The deterioration of AMD quality, including a drop in pH values, is
often observed at the end of the wet season in South Korea and is
attributed to limited dilution of AMD, as the monsoon ends, in the
summer (Kim et al., 2003).
The reactors quickly developed reducing conditions during the
acclimation period, yielding Eh values of nearly þ210 mV, at the
first sampling. The Eh steadily decreased thereafter until day 80
and remained relatively constant reaching values as low
as 147 mV (on day 117, #3) or 158 mV (on day 145, #1), before it
began to rise and approach the initial values at the end of the
154 H. Song et al. / Journal of Environmental Management 111 (2012) 150e158
experiment. The increase in Eh toward the end would suggest the
gradual inactivation of SRB as indicated by the increase of sulfate
concentrations over the same period (Fig. 3). Eh values were also
consistent with DO concentrations that were maintained at around
0 mg L1 up to day 152 but increased to up to 2.4 mg L1 on day 174.
The conductivity, TDS, and salinity, which are greatly influenced
by concentrations of metals and sulfate leached from substrate
materials, showed very similar breakthrough patterns throughout
the experiment, consistent with previously reported behavior in
a pilot-scale test (Guo et al., 2001). The peak values of the three
parameters were recorded in a mushroom compost-based reactor
(#3) on day 27 (9.1 mS cm1, 4.9 g L1, and 5.1& of conductivity,
TDS, and salinity, respectively). The bioreactors containing mixed
substrates consistently yielded lower values than the mushroom
compost reactor up to day 80. Hence, the data suggested that the
mushroom compost released higher amounts of ionic components
into the solution relative to the reactors #1 and #2 that also washed
out metals, giving rise to a peak of their concentration in the
effluent, but to a lower extent.
3.2.2. Sulfate, sulfide, DOC, and SRB counts
The mushroom compost released a significant amount of
sulfate, yielding higher concentrations of sulfate in the early reac-
tion times than the AMD source, especially for the reactor with
100% mushroom compost (Fig. 3). In a separate analysis, acid
soluble sulfate, determined as per Sobek et al. (1978), showed
a value of 27.5 g kg1 in mushroom compost. Such a release of
sulfate from mushroom compost was also reported in a previous
study (31.1 g kg1; Guo et al., 2001). The potential source of sulfate
in the mushroom compost could be either cow manure or gypsum
(CaSO42H2O), or both, since the former is a base material of
mushroom compost and the dissolution of the latter can be
enhanced through an ion-pair mechanism and the common ion
effect (Truesdale, 2011). In a previous study, as much as 7 g kg1 of
sulfate was found to originate from a composted poultry manure
Fig. 3. Sulfate reduction evolution during AMD treatment in pilot-scale bioreactors. Reactors #1 and #2: mushroom compost (20%), cow manure compost (10%), sawdust (20%), rice
straw (10%), creek sediment (20%), and limestone (20%); Reactor #3: mushroom compost (100%).
substrate (Zagury et al., 2006). However, after the initial leaching
out of sulfate from the organic substrates, the system was able to
consistently reduce sulfate concentrations in all three reactors
below the values in the AMD within 50 days, after which it
remained relatively stable until day 125, except for reactor #2. The
average sulfate removal within this period was of 40, 48, and 20%
for reactors #1, #2, and #3, respectively. The highest sulfate
removal in the reactors with mixed substrates (#1 and #2) was
observed on day 49, yielding a peak of 78 and 67% removals,
respectively. Meanwhile, the sulfate level in the mushroom
compost reactor (#3) was consistently higher than those in the
mixed substrate reactor until day 125, with its greatest removal
observed on the same day (47%). At the same time, the highest
concentrations of produced sulfides were recorded on day 125,
ranging from 26 (#3) to 51 mg L1 (#2). Sulfides were measurable
from day 14 onward, and their concentrations were generally
higher in the reactors with mixed substrates than in the mushroom
compost reactor, consistent with the greater sulfate reduction in
the mixed substrate reactors.
After day 125, sulfate and ferrous iron concentrations in AMD
significantly increased, and this deterioration in input water quality
had an adverse impact on the biological activity, leading to
increases in sulfate concentrations and decreases in sulfide
concentrations and SRB counts (especially in reactor #2). Never-
theless, in the most efficient reactor (#1), the sulfate level remained
well below that of the AMD throughout the entire experiment. DOC
concentrations gradually decreased with time to reach final values
of 5.2 (#1), 9.4 (#2), and 3.6 mg L1 (#3), with the exception of an
abnormally high value measured in reactor #2 on day 91
(609 mg L1). Apparently, DOC for all substrates was leached out in
a similar manner. These results suggest that a substrate with a high
initial DOC is likely to leach out a significant amount of DOC to the
effluent, losing available substrate for microbial growth. Conse-
quently, after some period of operation, the activity of anaerobic
bacteria in the reactor could become limited by the lack of
 H. Song et al. / Journal of Environmental Management 111 (2012) 150e158
substrate. SRB counts in reactor #3 are characterized by the highest
values at early time, followed by a sharp drop from 6-log to 3-log,
within three weeks (from day 125 to day 146). Wide variations in
SRB counts were also observed in reactor #2, whereas reactor #1
was the steadiest.
3.2.3. Total Fe, Fe2þ, Al, and Mn
The major mechanisms involved in metal removal in passive
bioreactors have been reported to be sorption and precipitation,
with some minor contribution from filtration through precipitate
formation (Neculita et al., 2007). The sorption saturation of
mushroom compost during the treatment of synthetic AMD (pH
3.5, 48 mg L1 Fe, and 22 mg L1 Mn) under high flow rates could be
reached in as few as 15 days (Stark et al., 1994), after which metal
removal is dominated by precipitation. In the present study, metal
removal efficiencies were consistent with the results from batch
bioreactor testing (Neculita et al., 2011). They showed the following
order: Al > Fe > Mn. Exceptions were recorded on two occasions,
which were related to the mushroom compost-based reactor, in
which Al concentrations in the effluents were always less than
5 mg L1, despite the concentration variation in AMD from 42.5
(day 125) to 172.5 mg L1 (day 49) (Fig. 4). There was a large
variation in Fe2þ concentration in the AMD, showing a trend of
gradual increase during the testing period. The increase in Fe2þ in
the AMD presumably stemmed from the seasonal effect that
induced enhanced photo-reduction of ferric hydroxide by sunlight
radiation during the summer season (McKnight and Duren, 2004).
The peak of Fe2þ in AMD was recorded on day 158 (86 mg L1),
which corresponded to the beginning of the dry season. The
concentrations of Fe2þ in the treated effluents were similar or
greater than those in AMD until day 49, but became lower after-
ward with the peak concentrations on day 158 in the mixed
substrate reactors (26 mg L1 in #1 and 59 mg L1 in #2). The
Fig. 4. Metal removal evolution during AMD treatment in pilot-scale bioreactors. Reactors #1 and #2: mushroom compost (20%), cow manure compost (10%), sawdust (20%), rice
straw (10%), creek sediment (20%), and limestone (20%); Reactor #3: mushroom compost (100%).
155
abrupt increase of Fe2þ in reactor #2 on day 158 coincides with the
sharp increase of sulfate and decrease of soluble sulfide concen-
trations occurring on the same day. While this observation is a clear
indication of the deterioration in performance of the reactor, it also
suggests that the formation of metal sulfide was the major pathway
of iron removal in the reactor.
Total Fe concentrations in the AMD ranged from 51 (day 23) to
235 mg L1 (day 49), with no clear trend in the temporal varia-
tion. One notable observation in the changes of total Fe concen-
tration was a significant increase in reactor #2 toward the end of
the reaction period (Fig. 4). This could be partially explained by
the reactor failure as reflected by the release of significant
amounts of Fe2þ and sulfate, and the sharp decrease of effluent
pH in the reactor. Furthermore, such changes in the geochemistry
within the reactor possibly re-dissolve precipitated or adsorbed
iron minerals and leach out a significant amount of iron, even
exceeding the level in AMD source water. Nevertheless, from day
49 to the end of testing, in reactors #1 and #3 (or up to day 158 in
reactor #2), satisfactory Fe removal was found, in the range of
68e92%, including day 49, when the total Fe concentration in
AMD was the highest (235 mg L1). Meanwhile, Mn concentra-
tions in input water varied from 13.8 (day 125) to 40.5 mg L1
(day 91), while in treated water it varied from 8.7 (#2 on day 125)
to 47.4 mg L1 (#3 on day 158) (Fig. 4). Overall, the effluent
quality of the mushroom compost-based reactor was even poorer
than the AMD over the first two months because of leaching of
sulfate and major cations. Mn release from this substrate material
led to higher concentrations in the treated effluent than AMD
during early reaction times. Similar results, i.e. marginal to very
low Mn removal efficiencies, were obtained in laboratory
column-bioreactors filled with Korean mushroom compost and
operated at 20-d of HRT and a Fe/Mn ratio of 10 (Chang et al.,
2000).
156 H. Song et al. / Journal of Environmental Management 111 (2012) 150e158
3.2.4. Na, K, Ca, and Mg
The significance of measuring these ions was, apart from
ensuring the micronutrients are present to sustain SRB activity, to
assess the influence of treatment system on water chemistry.
Previously reported results showed that passive biological treat-
ment systems (e.g. wetlands) could act as sink of Na, K, Ca and Mg
from inputs of acid mine drainage (Wieder and Lang, 1984).
The bioreactors released large amounts of Na during early
reaction times, as indicated by significantly higher Na concentra-
tions in the effluent as compared to AMD. Such a release of Na was
more apparent in the mushroom compost reactor, indicating the
major source of Na was mushroom compost (Fig. 5). Na concen-
trations dropped quickly after the initial leaching phase before they
increased again on day 91, which could be attributable to the
temporal Na shock in AMD source water that occurred on day 34
and 49. A similar leaching effect was also observed for K, but its
concentration in all reactors dropped to below 30 mg L1 after day
49 since the K concentration in AMD was constantly maintained at
a very low level.
Ca concentrations in the AMD varied between 176 and
420 mg L1, with less fluctuation than Mg, which ranged between
95 and 274 mg L1. The concentrations of Ca and Mg in the reactors
varied considerably during the reaction period, but in general, the
Ca concentrations remained higher than those in the AMD, while
no such observation was obvious for Mg except early on in the
reaction time. This indicates that the substrate materials contrib-
uted to the overall amount of Ca to some extent throughout the
reaction period, but the release of Mg was rather short-lived.
Increase in Ca concentrations is likely (and mainly) due to lime-
stone dissolution. A notable observation in the Ca and Mg profiles
was the sharp increase of concentrations between days 124 and
158, which may be related to a 0.6 unit decrease of the pH in AMD
occurring over the same period (Fig. 3). In other words, the
Fig. 5. Micronutrient evolution during acid mine drainage (AMD) treatment in pilot-scale bioreactors. Reactors #1 and #2: mushroom compost (20%), cow manure compost (10%),
sawdust (20%), rice straw (10%), creek sediment (20%), and limestone (20%); Reactor #3: mushroom compost (100%).
deterioration in AMD quality strongly affected the condition of all
reactors and led to the release of high concentrations of Ca and Mg,
especially in reactor #2.
3.2.5. Fe/Mn ratio effect on Mn removal efficiency
In addition to the possible reasons provided above for the very
poor Mn removal in passive bioreactors, the Fe/Mn ratio in fed
water may also play an important role in Mn removal within
bioreactors (Skousen et al., 2000). Mn is ubiquitous in AMD-
impacted waters, but it is one of the most problematic metals
since it does not readily form sulfidic minerals and requires high pH
(>8) for chemical oxidation to the less soluble Mn(IV) form
(Mariner et al., 2008). At pH <8, the kinetics of Mn oxidation are
very slow, requiring a secondary polishing treatment of bioreactor-
treated waters (Mariner et al., 2008). Some studies that reported
successful Mn removal to date were performed with a treated
effluent with low Fe concentrations (Mariner et al., 2008), or with
water with high Mn concentrations (up to 567 mg L1; Cheong
et al., 2010), i.e. a Fe/Mn ratio <2. The Fe/Mn ratio in previous
studies varied from 0, i.e. AMD did not contain Fe (Edwards et al.,
2009; Karathanasis et al., 2010) to 0.8 (Cheong et al., 2010) and
up to 10 (Chang et al., 2000).
In the present study, the Fe/Mn ratio in AMD water varied from
1.7 to 3.5 within the first 104 days, except on day 76 (4.8) and day
49 (6.2) (Fig. 6). Better removal of Mn was observed during this
period, with the highest removal efficiencies of 49% (#1) to 61%
(#2). After day 104, the Fe/Mn ratio increased progressively up to 8,
and Mn concentrations in treated water increased up to twice the
original concentrations in AMD, regardless of the type of substrate
material. In some cases where the bioreactor effluent had a higher
level of Mn than that of source AMD (e.g. day 24), the excess Mn
appears to originate from the reductive dissolution of solid Mn-
bearing minerals or of Mn complexes associated with substrate
 H. Song et al. / Journal of Environmental Management 111 (2012) 150e158
Fig. 6. Efficiency of Mn removal in relation to the Fe/Mn ratio from AMD in pilot-scale
bioreactors. Reactors #1 and #2: mushroom compost (20%), cow manure compost
(10%), sawdust (20%), rice straw (10%), creek sediment (20%), and limestone (20%);
Reactor #3: mushroom compost (100%).
materials (Fig. 6). The sharp increases of Mn in the reactor effluents
observed toward the end of the reaction time presumably resulted
from the dissolution of Mn-sulfides in response to a pH decrease of
source AMD.
Very optimistic results generated in small scale experiments
(Cheong et al., 2010) have not yet been confirmed in larger scales of
practical significance, especially at a higher Fe/Mn ratio and under
continuous flow. In fact, laboratory column-reactors tested for
short periods of time (less than 6 months) do not provide pertinent
data applicable to field-scale treatment systems. As a leading
example, the chitinous materials, which apparently seemed to be
the most promising substrate for Mn retention in laboratory
passive systems (Robinson-Lora, 2009), failed to provide consistent
results at a larger pilot-scale (Reisman et al., 2009).
4. Conclusions
The efficiency of a reactive mixture (in duplicate) comprised of
four organic substrates (cow manure compost, mushroom compost,
sawdust, and rice straw) was evaluated relative to mushroom
compost alone (control), in three pilot-scale passive bioreactors fed
for 174 days with AMD from a closed South Korean mine site.
Results indicated that mixed-substrates could yield comparable
performance to mushroom compost for reducing sulfate and
removing metals from AMD, while providing better effluent quality
in an initial phase of reaction (1e2 months). Deterioration of AMD
quality, which occurred after 100 days of operation, presumably
due to the reduced dilution of AMD with the onset of the dry
season, led to performance deterioration of all reactors, but the
reactors slowly recovered the treatment efficiencies toward the end
of testing. The reactors were effective for pH neutralization and
metal removal, showing a relative removal efficiency of
Al > Fe > Mn. However, the mushroom compost reactor released
significant amounts of sulfate and major cations that impaired the
effluent quality at early reaction times and reduced overall treat-
ment efficiency. Nevertheless, after the initial leaching out of
sulfate from the organic substrates, the system was able to reduce
sulfate concentrations in all three reactors below the values in the
AMD within 50 days, after which it remained relatively stable until
157
day 125. Furthermore, DOC loss from mushroom compost was
more significant than the mixed substrates, suggesting that
performance limitation due to a shortage of available organic
carbon could occur sooner in mushroom compost reactors. Satis-
factory Mn removal was achieved when the Fe/Mn ratio in the
solution was small (3.5), regardless of the substrates used. The
overall results of this study suggested that the use of mixed
substrates could be a viable option to overcome the limitations of
mushroom compost during AMD treatment.
Acknowledgments
This work was primarily supported by the KIGAM annual
research fund (project 10-3411) and, in part, by the Mine Recla-
mation Corporation Research Foundation. Any opinions, findings,
conclusions, or recommendations expressed are those of the
authors and do not necessarily reflect the views of the funding
agencies.
References
APHA, 2005. Standard Methods for the Examination of Water and Wastewater, 21st
ed. American Public Health Association, Washington, DC.
ASTM, 1995. Standard Test Method for pH of Soils. D4972e95a. ASTM International,
West Conshohocken, PA.
ASTM, 2009. Standard Methods for Sulfate-Reducing Bacteria in Water and Water-
Formed Deposits. D4412-84. ASTM International, West Conshohocken, PA.
Burdige, D.J., Nealson, K.H., 1986. Chemical and microbiological studies of sulfide-
mediated manganese reduction. Geomicrobiol. J. 4, 361e387.
Chang, I.S., Shin, P.K., Kim, B.H., 2000. Biological treatment of acid mine drainage
under sulphate-reducing conditions with solid waste materials as substrate.
Water Res. 34, 1269e1277.
Cheong, Y.W., Das, B.K., Roy, A., Bhattacharya, J., 2010. Performance of a SAPS-based
chemo-bioreactor treating acid mine drainage using low-DOC spent mushroom
compost, and limestone as substrate. Mine Water Environ. 29, 217e224.
Dvorak, D.H., Hedin, R.S., Edenborn, H.M., McIntire, P.E., 1992. Treatment of metal-
contaminated water using bacterial sulfate reduction: results from pilot-scale
reactors. Biotechnol. Bioeng. 40, 609e616.
Edwards, J.D., Barton, C.D., Karathanasis, A.D., 2009. A small-scale sulfate-reducing
bioreactor for manganese removal from a synthetic mine drainage. Water Air
Soil Pollut. 203, 267e275.
Guo, M., Chorover, J., Rosario, R., Fox, R.H., 2001. Leachate chemistry of
fielddweathered spent mushroom substrate. J. Environ. Qual. 30, 1699e1709.
Ji, S.W., Kim, S.J., Ko, J., 2008. The status of the passive treatment systems for acid
mine drainage in South Korea. Environ. Geol. 55, 1181e1194.
Karam, A., 1993. Chemical properties of organic soils. In: Carter, M.R. (Ed.), Soil
Sampling and Methods of Analysis. Lewis Publishers, Boca Raton, FL,
pp. 459e471.
Karathanasis, A.D., Edwards, J.D., Barton, C.D., 2010. Manganese and sulfate removal
from a synthetic mine drainage through pilot scale bioreactor batch experi-
ments. Mine Water Environ. 29, 144e153.
Kim, J.J., Kim, S.J., Choo, C.O., 2003. Seasonal change of mineral precipitates from the
coal mine drainage in the taebaek coal field, South Korea. Geochem. J. 37,
109e121.
Mariner, R., Johnson, D.B., Hallberg, K.B., 2008. Characterization of an attenuation
system for the remediation of Mn(II)contaminated waters. Hydrometallurgy 94,
100e104.
McKnight, D., Duren, S., 2004. Biogeochemical processes controlling midday ferrous
iron maxima in stream waters affected by acid rock drainage. Appl. Geochem.
19, 1076e1077.
MIRECO (Mine Reclamation Corporation), 2011. Yearbook of MIRECO Statistics.
available in Korean at. http://www.mireco.or.kr.
Neculita, C.M., Zagury, G.J., 2008. Biological treatment of highly contaminated acid
mine drainage in batch reactors: long-term treatment and reactive mixture
characterization. J. Hazard. Mater. 157, 358e366.
Neculita, C.M., Zagury, G.J., Bussière, B., 2007. Passive treatment of acid mine
drainage in bioreactors using sulfate-reducing bacteria: critical review and
research needs. J. Environ. Qual. 36, 1e16.
Neculita, C.M., Zagury, G.J., Bussière, B., 2008. Effectiveness of sulphate-reducing
passive bioreactors for treatment of highly contaminated acid mine drainage:
I. Effect of hydraulic retention time. Appl. Geochem. 23, 3442e3451.
Neculita, C.M., Yim, G.J., Lee, G.Y., Ji, S.W., Jung, J.W., Park, H.S., Song, H., 2011.
Comparative effectiveness of mixed organic substrates to mushroom compost
for treatment of mine drainage in passive bioreactors. Chemosphere 83, 76e82.
Reisman, D., Rutkowski, T., Smart, P., Gusek, J., Sieczkowski, M., 2009. Passive
treatment and monitoring at the standard mine superfund site Crested Butte,
CO. In: Barnhisel, R.I. (Ed.), Revitalizing the Environment: Proven Solutions and
Innovative Approaches. ASMR, Lexington, KY, pp. 1107e1128.
158 H. Song et al. / Journal of Environmental Management 111 (2012) 150e158
Robinson-Lora, M.A., 2009. Evaluation of Chitinous Materials as a Multifunctional
Substrate for the Remediation of Mine Impacted Water. PhD dissertation,
Pennsylvania State University, State College, PA, 133 pp.
Skousen, J.G., Sexstone, A., Ziemkiewicz, P.F., 2000. Acid mine drainage control and
treatment. In: Hartfield, J.L., Volenec, J.G., Dick, W.A. (Eds.), Reclamation of
Drastically Disturbed Lands, pp. 131e169.
Sobek, A.A., Schuller, W.A., Freeman, J.R., Smith, R.M., 1978. Field and Laboratory
Methods Applicable to Overburdens and Mine Soils, EPA-600/2-78-054, 60e62.
Washington, DC.
Song, H., Yim, G.J., Ji, S.W., Nam, I.H., Neculita, C.M., Lee, G.Y., 2012. Performance of
mixed organic substrates during the treatment of acidic and moderate mine
drainage in column bioreactors. ASCE J. Environ. Eng. 138. http://dx.doi.org/
10.1061/(ASCE)EE.1943-7870.0000567.
Stark, L.R., Williams, F.M., 1994. The roles of spent mushroom substrate for the
mitigation of coal mine drainage. Compost Sci. Util 2, 84e94.
Stark, L.R., Williams, R.W., Williams, F.M., 1994. Restoring the capacity of spent
mushroom compost to treat coal mine drainage by reducing the inflow rate:
a microcosm experiment. Water Air Soil Pollut. 75, 405e420.
Stone, A.T., Morgan, J.J., 1984. Reduction and dissolution of manganese(III) and
manganese(IV) oxides by organics: 2. Survey of reactivity of organics. Environ.
Sci. Technol. 18, 617e624.
Stumm, W., Morgan, J.J., 1996. Aquatic Chemistry, Chemical Equilibria and Rates in
Natural Waters, third ed.. John Wiley & Sons, New York.
Thomas, R.C., Albano, C., Bays, J.S., 2010. Advances in passive treatment
of mine impacted water. In: Proc. of the 12th IWA Int. Conf. Wetland
Syst. Water Pollut. Control. Venice, Italy, October 4e8, 2010, vol. II, pp.
975e982.
Truesdale, V.W., 2011. Rate equations and an ion-pair mechanism for batch disso-
lution of gypsum: repositioning the shrinking object model at the core of
hydrodynamic modeling. Aquat. Geochem. 17, 141e164.
Venot, C., Figueroa, L., Brennan, R.A., Wildeman, T.L., Reisman, D.,
Sieczkowski, M., 2008. Comparing chitin and organic substrates on the
national tunnel waters in Blackhawk, Colorado for manganese removal. In:
Barnhisel, R.I. (Ed.), New Opportunities to Apply Our Science. ASMR, Lex-
ington, KY, pp. 1352e1366.
Villinski, J.E., Saiers, J.E., Conklin, M.H., 2003. The effects of reaction-product
formation on the reductive dissolution of MnO2 by Fe(II). Environ. Sci. Tech-
nol. 37, 5589e5596.
Waybrant, K.R., Ptacek, C.J., Blowes, D.W., 2002. Treatment of mine drainage using
permeable reactive barriers: column experiments. Environ. Sci. Technol 36,
1349e1356.
Wieder, R.K., Lang, G.E., 1984. Influence of wetlands and coal mining on stream
water chemistry. Water Air Soil Pollut. 23, 381e396.
Zagury, G.J., Kulnieks, V., Neculita, C.M., 2006. Characterization and reactivity
assessment of organic substrates for sulfate-reducing bacteria in acid mine
drainage treatment. Chemosphere 64, 944e954.