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KSCE Journal of Civil Engineering (2013) 17(4):718-728

DOI 10.1007/s12205-013-0149-8

Geotechnical Engineering

www.springer.com/12205

Improvements in Engineering Properties of Soils through


Microbial-Induced Calcite Precipitation
Ng Wei Soon*, Lee Min Lee**, Tan Chew Khun***, and Hii Siew Ling****
Received March 22, 2012/Revised May 25, 2012/Accepted July 12, 2012

Abstract
Microbial-Induced Calcite Precipitation (MICP) has recently emerged as a sustainable technique for soil improvement. This paper
aims to study the effectiveness of MICP in improving the shear strength and reducing the hydraulic conductivity of soils. A species of
Bacillus group, B. megaterium was used to trigger the calcite precipitation. The experimental variables included soil types (tropical
residual soil and sand), soil densities (85%, 90%, and 95% of their respective maximum densities), and treatment conditions
(untreated, treated with cementation reagents only, treated with B. megaterium only, and treated with B. megaterium and cementation
reagents). The results showed that MICP could effectively improve shear strength and reduce hydraulic conductivity for both residual
soil and sand. The improvements, however, varied with soil densities, soil types, and treatment conditions. With MICP treatment, the
improvement ratios in shear strength of the residual soil specimens were significantly higher (1.41-2.64) than those of the sand
specimens (1.14-1.25). On the contrary, the sand specimens resulted in greater hydraulic conductivity reduction ratios (0.09-0.15)
than those of the residual soil specimens (0.26-0.45). These observations can be explained by the particle-particle contacts per unit
volume and pore spaces in the soil specimens. Both soil specimens when treated with cementation reagents only exhibited slight
alterations in the shear strength (ranging from 1.06-1.33) and hydraulic conductivity (ranging from 0.69-0.95). The results implied
that natural calcite forming microorganisms only exist for insignificant amount. The amount of calcite precipitated in the treated
residual soil specimens ranged from 1.080% to 1.889%. The increments of calcite content in the treated sand specimens were
comparatively higher, ranging from 2.661% to 6.102%. The results from Scanning Electron Microscope (SEM) analysis confirmed
the experimental findings.
Keywords: microbial-induced calcite precipitation, soil improvement, shear strength, hydraulic conductivity, B. megaterium

1. Introduction
Nowadays, construction on problematic soils is inevitable
owing to the growing scarcity of land worldwide. Problematic
soils are commonly characterized by low strength and high
compressibility (Ho and Chan, 2011; Huat, 2006; Kazemian et
al., 2011). In tropical regions like Malaysia, soils are subject to
further softening due to infiltration of intense and prolonged
downpours. Consequently, development on the problematic soils
is highly susceptible to severe geohazards including excessive
settlement of embankment or foundation, debris flow, and
catastrophic landslide.
Studies on the soil improvement techniques can be found in
abundance. The important features of soil improvement include:
improving the shear strength of soil, reducing the potential for
total and differential settlement, reducing the time during which

the settlement takes place, reducing the potential for liquefaction


in saturated fine sand or hydraulic fills, reducing the hydraulic
conductivity of soil, removing or excluding water from the soil
etc (Kazemian and Huat, 2009; Krebs and Walker, 1971;
Leonards, 1962). Conventionally, the norm is to replace low
strength soil deposits with engineering fill. Presently, the use of
chemical grouting is becoming increasingly popular owing to its
economical benefit. Chemical grouting can be achieved with a
variety of additives including Portland cement, lime, asphalt,
sodium silicate, acrylate, lignin, urethane, and resins. While many
of these additives have proven successful (Anagnostopoulos and
Hadjispyrou, 2004; Basha et al., 2005; Karol, 2003; Peethamparan
et al., 2009; Xanthakos et al., 1994), the additives often modify
the pH of soils, and may contaminate the soils and groundwater
(Dejong et al., 2006; Karol, 2003).
In recent years, with increasing awareness of environmental

*Postgraduate Candidate, Faculty of Engineering and Science, Universiti Tunku Abdul Rahman, Kuala Lumpur 53300, Malaysia (E-mail: just_ws@hotmail.com)
**Assistant Professor, Faculty of Engineering and Science, Universiti Tunku Abdul Rahman, Kuala Lumpur 53300, Malaysia (Corresponding Author, E-mail:
mllee@utar.edu.my)
***Assistant Professor, Faculty of Engineering and Green Technology, Universiti Tunku Abdul Rahman (Perak Campus) 31900, Malaysia (E-mail: tckhun@utar.edu.my)
****Associate Professor, Faculty of Engineering and Science, Universiti Tunku Abdul Rahman, Kuala Lumpur 53300, Malaysia (E-mail: hiisl@utar.edu.my)
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Ng Wei Soon, Lee Min Lee, Tan Chew Khun, and Hii Siew Ling

issues, there has been a remarkable shift toward green and


sustainable technologies. As all chemical grouts except sodium
silicate are toxic and hazardous, there are expressed concerns
over their use for soil improvement (Dejong et al., 2010). A new
soil improvement technique that utilizes a biological process, which
is termed technically as MICP, has emerged recently. MICP has
been enabled through interdisciplinary researches at the confluence
of microbiology, geochemistry, and geotechnical engineering, to
find natural treatments for soil improvement (Dejong et al., 2010).
MICP is a biological process occurs in nature. It is intensified by
introducing a large population of urease-producing microorganisms
and cementation reagents into the soil matrix, whereby a cement
compound is generated to improve the engineering properties of the
soil. The environmental friendly characteristics of the ureaseproducing microorganisms will cause very little, if not no
impairment to the soil, human health, and environment.
Despite of the fact that the MICP soil improvement technique
is still a relatively young technology, many studies pertaining to
the topic have been reported including Baveye et al. (1998),
Castainer et al. (1999), Ehrlich (1999), Mitchell and Santamarina
(2005), Lian et al. (2006), Ivanov and Chu (2008), Dejong et al.
(2010), Okwadha and Li (2010), Harkes et al. (2010) and, Lu et
al. (2010). Dejong et al. (2006) performed laboratory experiments
to investigate the MICP on loose, collapsible sand specimens
using B. pasteurii. They found that MICP-treated specimens
exhibited a noncollapsing strain softening shear behavior, with
higher initial shear stiffness and ultimate shear capacity than
untreated loose specimens. Baskar et al. (2006) isolated the
calcite-forming microorganisms, B. thuringiensis and B. pumilis
from stalactites, sampled from three caves in Sahastradhara,
Dehradun, India. They concluded that microbial activity and
temperature are the key factors promoting calcite precipitation.
Harkes et al. (2010) researched on the methodology to distribute
and fix B. pasteurii homogeneously in sand bed. They found that
the homogeneous distribution can be achieved by performing twophase injection procedures: injection of a bacterial suspension into
the sand body, immediately followed by injection of a fixation fluid
of high salt content. Ivanov and Chu (2008) provided a detailed
review on MICP applications for soil improvement. Although there
are diverse potential applications of MICP, they concluded that, at
present, promising applications are only focusing on bioclogging
and biocementation. Biocementation is defined as the improvement
of soil strength by the production of particle-binding materials
through microbial means, while bioclogging is the reduction of
hydraulic conductivity of soil or porous rock by pore-filling
materials generated by microbial processes.
Most Bacillus strains can precipitate calcite through the
conversion of urea into ammonia and carbon dioxide (Hammes
et al. 2003). The previous reported studies however, mainly
adopted B. pasteurii as the urease-producing microorganism to
consolidate loose sand columns. Studies which employed other
Bacilli and different soil types are still very limited. The present
experimental work investigates the effectiveness of a different
Bacillus strain, B. megaterium, in improving the shear strength

(biocementation) and reducing the hydraulic conductivity


(bioclogging) of two soil types, i.e., tropical residual soil and sand.

2. Testing Materials
2.1 Soil Specimens
The two types of soil employed in this study were tropical
residual soil and sand. The tropical residual soil specimen was
obtained from a site in Universiti Tunku Abdul Rahman, Kuala
Lumpur campus compound, while the sand specimen was of
typical concrete sand. Table 1 tabulates the values of physical
indices of the soil specimens obtained from the standard soil
properties tests. The soil particle size distributions are presented
in Fig. 1. Based on the British Standard soil classification
system, the tropical residual soil specimen was classified as
Sandy Silt with high plasticity, while the sand specimen was
classified as Well Graded Sand.
Standard proctor compaction test was performed to obtain the
compaction curve of the tropical residual soil. The Maximum
Dry Density (MDD) of the soil was found to be 1563 kg/m3. To
investigate the effectiveness of the MICP treatment for soils of
varying density, the residual soil specimens were compacted into
Table 1. Physical Properties of the Soil Specimens

Composition
Gravel (%)
Sand (%)
Silt (%)
Clay (%)
LL (%)
PL (%)
PI
Soil Classification BSCS
Maximum Dry Density (MDD)
Maximum Index Density (max)
Minimum Index Density (min)

719

Residual Soil
(Sandy Silt)

Sand

0
29
55
16
58.0
44.3
13.7
MHS
1563 kg/m3
-

2
96
2
0
SW
1842 kg/m3
1439 kg/m3

Fig. 1. Particle Size Distributions of the Soil Specimens


KSCE Journal of Civil Engineering

Improvements in Engineering Properties of Soils through Microbial-Induced Calcite Precipitation

three different densities, i.e., 85% of MDD, 90% of MDD, and


95% of MDD. For the sand specimen, the minimum (min) and
maximum (max) index densities were determined in compliance
with the procedures of ASTM D4254 (ASTM 2000) and ASTM
D4253 (ASTM 2000), respectively. Three densities were compacted
within the range of the minimum (min = 1439 kg/m3) and
maximum (max = 1842 kg/m3) index densities, i.e., 85% of max,
90% of max, and 95% of max.

residual soil specimens, the desired densities were achieved by


compacting the soils into prefabricated moulds with the moisture
content corresponding to the desired density determined from the
compaction curve. For the sand specimens, the desired densities
were achieved by tamping the predetermined amount of sand
specimens in layers. Upon compaction, all the soil specimens
were saturated to ensure steady flow of cementation reagents
through the soil specimens during the MICP treatment.

2.2 Microorganism
The urease-producing microorganism used in this study was B.
megaterium ATCC 14581. It is a rod-shaped, Gram positive soil
bacterium of diameter 2 m to 5 m. It can be found in diverse
environment from rice paddies to dried food, seawater,
sediments, fish, and even in bee honey (Wittmann et al., 2010).
The role of B. megaterium is to produce enzyme urease
through its metabolic activity under proper cultivation process.
The enzyme urease triggers the MICP biochemical reaction by
hydrolyzing urea (CO(NH2)2) through the following reaction:

3.2 MICP Treatments


As explained in the previous sections, MICP can be achieved
by introducing the urease-producing microorganisms into soil
together with the external sources of urea and calcium chloride
solution (Martinez et al., 2011; Stocks-Fischer et al., 1999;
Whiffin et al., 2007). The technology has been applied successfully
in diverse engineering fields including the consolidation of sand
columns, improvement of concrete strength, repairing the
concrete cracks etc (Achal et al., 2011; De Muynck et al., 2010;
Van Tittelboom et al., 2010).
The rate of calcite precipitation is an important process
variable. The rate should not be too high to avoid rapid
precipitation and consequent clogging near the inlet of the
specimen mould. The precipitation rate also should not be too
low as the specimen would take a long duration to cement (Van
Paassen et al., 2010).
To obtain a significant improvement in strength for loose
granular sand, at least 60 kg calcite per meter cube of soil has to
be precipitated. This corresponds to approximately 2 mol CaCO3
precipitate per liter of pore space (Van Paassen et al., 2010;
Whiffin et al., 2007). Qabany et al. (2011) found that this
amount of precipitated calcite can be achieved by injecting one
pore volume of 0.25 M CaCl2 / urea solution into the soil
specimen at intervals of 6 hours for a treatment period of 38-48
hours.
In the present study, B. megaterium was cultivated in nutrient
broth (pH 7.0 prior to autoclaving) at 130 rpm and incubation
temperature of 37oC. 13.0 g of nutrient broth was dissolved in
one liter of distilled water, which consisted of 5.0 g of peptone,
5.0 g of sodium chloride, 2.0 g of yeast extract, and 1.0 g of meat
extract. The culture with concentration of 5 107 cfu/mL
(optical density of 1.3) was then mixed with air-dried soil
specimens. The concentration of B. megaterium was determined
by the spread plate technique. Serial dilution was performed on
the incubated culture. The diluted culture was then spread on the
surface of agar plate where the colonies were counted after 16
hours of incubation. Subsequently, the soil mixture with the
addition of appropriate moisture content was compacted to the
desired densities.
Figures 2(a) and (b) show the schematic diagram of laboratory
setup and soil specimen mould used for the MICP treatment,
respectively. Prior to commencing the MICP treatment process,
the soil specimens were flushed through with a fixation fluid (50
mM CaCl2) to saturate the soil specimen and fix the B. megaterium
homogenously in soil specimen. The MICP treatment was

CO (NH2)2 + 3H2O 2NH4+ + HCO3 + OH

(1)

The ammonium (NH4+) increases the pH and causes the


bicarbonate (HCO3) to precipitate with calcium ion (Ca2+) from
the calcium chloride supplied in order to form the calcium calcite
(CaCO3):
Ca2+ + HCO3 + OH CaCO3 + H2O

(2)

The calcite generated is responsible for cementing and clogging


the soil specimens. Biocementation is achieved when the calcite
crystals precipitate on the surface or form bridges between the
existing soil grains. These calcite crystals bond the soil particles
and forbid movement of the grains, and hence enhance the
strength and stiffness properties of the soil (Harkes et al., 2010).
Bioclogging, on the other hand, is the formation of pore-filling
materials (calcite) through microbial process and subsequently
results in a reduction in soil porosity and hydraulic conductivity
(Ivanov and Chu, 2008).
2.3 Cementation Reagents
Cementation reagents serve as the raw materials for calcite
formation in the MICP process. The cementation reagents
employed in this study comprised 0.25 M solution of urea (CO
(NH2)2) and calcium chloride (CaCl2). The cementation reagents
also contained 3 g nutrient broth, 10 g ammonium chloride
(NH4Cl), and 2.12 g sodium bicarbonate (NaHCO3) per litre of
deionized water (Dejong et al., 2006; Qabany et al., 2011;
Stocks-Fischer et al., 1999; Stoner et al., 2005). All chemicals
used were of Analytical Reagent (AR) grade.

3. Methodology
3.1 Preparation of Soil Specimens
Prior to the MICP treatment, the soil specimens were air-dried
in the laboratory at room temperature for 1 week. For the
Vol. 17, No. 4 / May 2013

720

Ng Wei Soon, Lee Min Lee, Tan Chew Khun, and Hii Siew Ling

hours. The flow velocity was controlled at approximately 1.7


105 m/s. This was done by regulating the pressure of the air
compressor. The pressurized air compressed the reagent solution
in the pressure tank, and eventually created a hydraulic gradient
between the inlet and outlet of the soil specimen mould. This
treatment configuration was applied for all soil specimens.
3.3 Shear Strength and Hydraulic Conductivity Tests
Shear strength and hydraulic conductivity of the treated soil
specimens were tested in the laboratory. For the residual soil
specimens, the shear strength was determined by performing the
unconfined compression test on 50 mm diameter saturated
specimen. The hydraulic conductivity was determined by the
falling head permeability test. For the sand specimens, the shear
strength was determined by the standard direct shear box test on
60 mm 60 mm dry specimens. The hydraulic conductivity was
determined by the constant head permeability test. All test
procedures were in compliance with the British Standard 1377
(BSI, 1990). The same mould was used for the MICP treatment
and subsequent testing in order to ensure minimal disturbance on
the tested soil specimens.
Fig. 2. (a) Schematic Diagram of Laboratory Setup: (b) Soil Specimen Mould

performed by injecting one pore volume of 0.25 M CaCl2/urea


solution into the soil specimens at intervals of 6 hours over 48

3.4 Experimental Design


The experimental design focused mainly on the effects of soil
types (residual soil and sand), soil densities (85%, 90%, and 95%
of maximum density), and treatment conditions (untreated, treated
with B. megaterium only, treated with cementation reagents only,

Table 2 Experimental Design


No.
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
20.
21.
22.
23.
24.

Experiment abbreviation
0.95RU
0.95RM
0.95RR
0.95RT
0.90RU
0.90RM
0.90RR
0.90RT
0.85 RU
0.85RM
0.85RR
0.85RT
0.95SU
0.95SM
0.95SR
0.95ST
0.90SU
0.90SM
0.90SR
0.90ST
0.85SU
0.85SM
0.85SR
0.85ST

Soil type
Residual soil
Residual soil
Residual soil
Residual soil
Residual soil
Residual soil
Residual soil
Residual soil
Residual soil
Residual soil
Residual soil
Residual soil
Sand
Residual soil
Sand
Sand
Sand
Residual soil
Sand
Sand
Sand
Residual soil
Sand
Sand

Soil density
0.95 MDD
0.95 MDD
0.95 MDD
0.95 MDD
0.90 MDD
0.90 MDD
0.90 MDD
0.90 MDD
0.85 MDD
0.85 MDD
0.85 MDD
0.85 MDD
0.95max
0.95max
0.95max
0.95max
0.90max
0.90max
0.90max
0.90max
0.85max
0.85max
0.85max
0.85max
721

Treatment method
Untreated
B. megaterium only
Reagents only
B. megaterium & Reagents
Untreated
B. megaterium only
Reagents only
B. megaterium & Reagents
Untreated
B. megaterium only
Reagents only
B. megaterium & Reagents
Untreated
B. megaterium only
Reagents only
B. megaterium & Reagents
Untreated
B. megaterium only
Reagents only
B. megaterium & Reagents
Untreated
B. megaterium only
Reagents only
B. megaterium & Reagents
KSCE Journal of Civil Engineering

Improvements in Engineering Properties of Soils through Microbial-Induced Calcite Precipitation

and treated with B. megaterium and cementation reagents) on the


shear strength and hydraulic conductivity of soils. Each experiment
was repeated for three times, and the average of these three
values was considered for the shear strength and hydraulic
conductivity determinations.
The untreated soil specimens served as controls. The treatment
with cementation reagents only was used to investigate the
existence of naturally inhibited calcite forming microorganisms
in the soil specimens. The specimens treated with microorganism
only were used to monitor the effect of biomass on the shear
strength and hydraulic conductivity of the soils. The details of
the experimental design are tabulated in Table 2. A total of 144
laboratory experiments were performed to cater for the 24
combinations of soil specimens and treatment conditions.

4. Experimental Results

Fig. 4. Shear Strength Improvements of Residual Soil Specimens

4.1 Shear Strength of Residual Soil


Figure 3 compares the stress-strain curves of the untreated
residual soil specimens (RU) and those treated with B.
megaterium and cementation reagents (RT). The unconfined
compressive strength (qu) of the soil is defined as the peak stress
or the stress that yields 20% of axial strain, whichever is lower.
The shear strengths of the residual soils in this study were
characterized by the undrained shear strength parameter (cu),
which was taken as half of the unconfined compressive strength
(qu), i.e., cu = qu. Fig. 4 summarizes the shear strength results
of the residual soil specimens. The shear strength of MICPtreated residual soils was improved for all densities (0.85RT,
0.90RT, 0.95RT). The shear strength improvement ratio increased
with increased density, i.e., 1.41 (41%), 2.59 (159%) and 2.64
(164%) for specimens of 0.85RT, 0.90RT, and 0.95RT, respectively.
The soil specimens treated with cementation reagents only
(0.95RR, 0.90RR, and 0.85RR), also exhibited increased shear
strength. The undrained shear strength parameter improved by
1.11, 1.25 and 1.33 for the specimens of 0.85RR, 0.90RR, and
0.95RR, respectively. The results implied that MICP was triggered
by the microorganisms inhibiting naturally in the soil deposits.
The improvements (ranging from 1.11-1.33), however, were

Fig. 3. Stress-strain Curve for Residual Soil Specimens


Vol. 17, No. 4 / May 2013

lower compared to the specimens treated with B. megaterium


and cementation reagents (ranging from 1.41-2.64). This was
because the introduction of B. megaterium resulted in a higher
production of urease enzyme. The enzyme triggered more calcite
precipitation and led to greater enhancement in shear strength.
The results for the specimens treated with microorganism only, i.e.,
0.85RM, 0.90RM and 0.95RM were not shown in Fig. 4 because
no visible improvement in shear strength was observed in these
specimens. The results implied that biomass was ineffective in
improving the shear strength of the residual soil.
4.2 Hydraulic Conductivity of Residual Soil
Figure 5 shows the results of hydraulic conductivity for the
residual soil specimens. The saturated hydraulic conductivity
(ksat) of untreated residual soil specimens ranged between 1.0
107 m/s and 9.3107 m/s, in which the values were directly
proportional to the soil density. The saturated hydraulic conductivity

Fig. 5. Saturated Hydraulic Conductivity Reductions of Residual


Soil Specimens
722

Ng Wei Soon, Lee Min Lee, Tan Chew Khun, and Hii Siew Ling

of tropical residual soil typically lies in the range of 1106 to


1108 m/s (Tan et al., 2008).
The saturated hydraulic conductivity of MICP-treated soil was
markedly reduced for all densities. The reduction in hydraulic
conductivity inflicted by the calcite can be explicitly seen by
observing the margin between the saturated hydraulic conductivities
of the untreated specimens (0.85RU, 0.90RU and 0.95RU) and
those treated with B. megaterium and cementation reagents
(0.85RT, 0.90RT and 0.95RT). The greatest reduction in hydraulic
conductivity occurred in the densest specimen (0.95RT) where
the ratio of saturated hydraulic conductivity between treated and
untreated specimens was 0.26 (a reduction of 74%). The
reduction ratios for 0.90RT and 0.85RT specimens were 0.40
(60%) and 0.45 (55%), respectively.
Similar to the observations in shear strength, the residual soil
specimens treated with cementation reagents only (0.85RR,
0.90RR and 0.95RR) exhibited slight alteration in saturated
hydraulic conductivity (decreased by not more than 30% of
untreated). These observations confirmed the earlier finding that
a relatively small amount of calcite precipitating microorganism
exists naturally in the residual soil. Furthermore, the residual soil
specimens treated with microorganism only, without the supply
of cementation reagents (0.85RM, 0.90RM, and 0.95RM)
experiences no significant diminution in hydraulic conductivity.
In the nutshell, diminution in hydraulic conductivity of soil was
mainly inflicted by calcite and the effect was proportional to the
soil density. The formation of calcite clogged most of the pores
and reduced the saturated hydraulic conductivity of the residual
soil effectively.
4.3 Shear Strength of Sand
Figure 6 shows a sample of shear stress versus horizontal
displacement results obtained from the direct shear tests for the
untreated sand specimens (SU), and those treated with B.
megaterium and cementation reagents (ST). The shear strength
of the dry sand specimens used in this study was characterized by
the effective internal friction angle ( ') only. Fig. 7 summarizes
the improvement in the effective internal friction angle of the
sand specimens.
The effective internal friction angles of the untreated sand

Fig. 6. Stress-strain Curve for Sand Specimens

Fig. 7. Shear Strength Improvements of Sand Specimens

specimens were between 39.9o and 48.8o. The internal friction


angles of the MICP-treated sand specimens (0.85ST, 0.90ST and
0.95ST) were generally higher than the untreated specimens
(0.85SU, 0.90SU and 0.95SU). The 0.85ST specimen had the
greatest improvement ratio (1.25), followed by 0.90ST specimen
(1.17) and 0.95ST specimen (1.14). The improvement ratio
decreased with increased density. The trend was opposite to the
results observed in the residual soil specimens. Furthermore, the sand
specimens (1.14-1.25) exhibited significantly lower improvement
ratios than the residual soil specimens (1.41-2.64).
For the specimens treated with cementation reagents only
(0.85SR, 0.90SR and 0.95SR), the shear strength improvement
ratio was markedly lower (1.06-1.15) compared to the specimens
treated with B. megaterium and cementation reagents (1.141.25). The specimens treated with microorganism only (0.85SM,
0.90SM, 0.95SM) barely had any effect on the shear strength
alterations.
4.4 Hydraulic Conductivity of Sand
The saturated hydraulic conductivity (ksat) of the sand
specimens is illustrated in Fig. 8. The saturated hydraulic
conductivities of the untreated sand specimens were in the orders
of 104 to 103 m/s. These values are in close agreement with
typical saturated hydraulic conductivity of fine to medium sand
specimens (Brassington, 1988).
Similar to the trend observed in shear strength of sand
specimens, the reduction in hydraulic conductivity becomes less
effective with the increased density. The greatest reduction in
hydraulic conductivity occurred in the 0.85ST specimen in
which the hydraulic conductivity decreased by approximately
one order of magnitude from 3.5 103 m/s to 3.2 104 m/s (a
reduction ratio of 0.09). As the density of the specimen
increased, the reduction ratios of hydraulic conductivity were
marginally lesser as observed in 0.90ST (0.14) and 0.95ST
specimens (0.15).
For the sand specimens treated with cementation reagents only,

723

KSCE Journal of Civil Engineering

Improvements in Engineering Properties of Soils through Microbial-Induced Calcite Precipitation

4.5 Quantitative Analysis of Calcite Precipitated


The quantitative measurements of the carbonate content in the
soil specimens were carried out by gravimetric analysis using the
acid-treatment weight loss technique. Dry soil samples of 20 g
were prepared for the tests. The carbon dioxide deliberated from
the reaction between diluted hydrochloric acid (2 molar) and
carbonate in soil was indicated by the effervescence. The residue
was collected using a filter paper and oven-dried at temperature
of 105C. The measured weight loss of the soil sample was used
to estimate the percentage of carbonate content in the soil
specimens. It was assumed that the increment of carbonate
content in the soil specimens after MICP treatment was caused
by the formation of calcium carbonate.

The carbonate contents of the untreated residual soil and sand


were 0.670% and 0.274%, respectively. Figure 9 correlates the
carbonate content of the MICP-treated residual soil specimens
(0.85RT, 0.90RT and 0.95RT) with their shear strength and
hydraulic conductivity alterations. The carbonate contents of the
0.85RT, 0.90RT and 0.95RT soil specimens were found to be
1.750%, 2.559%, and 2.381%, respectively. By subtracting their
initial content in untreated specimens, the increments of calcite
contents after the MICP treatment were 1.080%, 1.889%, and
1.711%, respectively.
The 0.85RT specimen recorded the lowest carbonate content,
and hence contributed to the lowest alterations in both shear
strength and hydraulic conductivity. The highest carbonate
content was measured in the specimen of 0.90RT, followed by
the specimen of 0.95RT with a slightly lower carbonate content.
The overall trend of the carbonate content showed reasonably
good comparisons with the trends of the shear strength
improvement and hydraulic conductivity reduction. Dense soil
has a close arrangement of soil particles, and this contributes to
more inter-particle contact points per unit volume. The
precipitation of calcite at these inter-particles contact points
reduces the pore size (reduces hydraulic conductivity) and
improves the bonding between soil particles (improves shear
strength).
The correlations between the carbonate content of the MICPtreated sand specimens (0.85ST, 0.90ST and 0.95ST) and their
shear strength and hydraulic conductivity alterations are
demonstrated in Fig. 10. The calcite contents of the 0.85ST and
0.90ST specimens were almost identical to each other, i.e.,
6.376% and 5.943%, respectively. However, a significant drop of
carbonate content (2.935%) was observed in the 0.95ST
specimen. The dense sand (0.95ST) experiences low calcite
precipitation after the MICP treatment because it has a relatively
smaller pore space.
By comparing the amounts of calcite precipitated in both the
sand and residual soil specimens, it can be found that the soil
particle size has a significant effect on the calcite precipitation.
The amounts of calcite in the treated sand specimens (ranging
from 2.661% to 6.102%) were generally higher than those of the

Fig. 9. Correlations between Carbonate Content and Alterations in


Shear Strength and Hydraulic Conductivity of Residual Soil
Specimens

Fig. 10. Correlations between Carbonate Content and Alterations


in Shear Strength and Hydraulic Conductivity of Sand Specimens

Fig. 8. Saturated Hydraulic Conductivity Reductions of Sand


Specimens

the reductions in hydraulic conductivity were negligible. The


hydraulic conductivities of the 0.85SR, 0.90SR and 0.95SR
were only reduced by an average of 7%. Similarly, the
reductions in hydraulic conductivity of sand specimens treated
with microorganism only (0.85SM, 0.90SM, and 0.95SM) were
also negligible.

Vol. 17, No. 4 / May 2013

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Ng Wei Soon, Lee Min Lee, Tan Chew Khun, and Hii Siew Ling

residual soil specimens (ranging from 1.750% to 2.559%). This


observation can be explained by the pore spaces available in the
soils. For the residual soil, some portions of the soil particles
were smaller than 2 m. As the bacteria used in this study has a
diameter of 2 m to 5 m, the limited pore spaces controlled the
passage of bacteria through the soil, and hence lower calcite
amount was precipitated. The large pore space in sand can
accommodate for more bacteria and reagent solutions, and allow
for more calcite precipitation.
From the Fig. 10, it is apparent that the overall trend of the
calcite content showed good agreement with the trends of
improvement in shear strength and reduction in hydraulic
conductivity of the treated sand specimens. It should be noted
that despite of significantly higher calcite content precipitated in
sand as compared to the residual soil, the improvements in shear
strength of sand specimens were still much lower than those of
residual soil specimens. The reason could be the calcite was not
formed at the inter-particle contact points in view of large pore
size in sand. Whiffin et al. (2007) found that calcite content of
above 3.5% was effective in improving the shear strength of
sand. However, the present experimental results showed that the
shear strength was not improved considerably despite calcite
content of 6% had been produced. The possible explanation of
this discrepancy is that the sand specimen used in the study of
Whiffin et al. (2007) was finer than the sand adopted in the
present study. For example, the median grain size (D50) of the
sand adopted in the study of Whiffin et al. (2007) was 165 m,
which was much finer than the D50 of 520 m adopted in the
present study. The results suggested that the shear strength of
soils with fine grain size could be effectively improved at lower
calcite content due to the dominant role of inter-particle contact
points per unit volume. Nevertheless, there was still a great
reduction in hydraulic conductivity due to the high calcite
contents in treated sand. The precipitated calcite caused clogging
at the sand pore spaces and pore throats.
4.6 Scanning Electron Microscopy Analysis
Scanning Electron Microscope (SEM) imagery was performed
on the selected soil specimens to visualize the calcite precipitation
in the soil specimens. The SEM images were captured using the

Fig. 11. SEM Images for the Residual Soil Specimens: (a) Untreated,
(b) Treated with Cementation Reagents Only, (c) Treated with
B. megaterium and Cementation Reagents

Hitachi S-3400N Scanning Electron Microscope. The tests were


of particular interest on the formation of calcite crystals upon
MICP treatment. Fig. 11 shows the SEM images of the residual
soil specimens treated under three different conditions, i.e.
untreated, treated with cementation reagents only, and treated
with B. megaterium and cementation reagents. Relatively smooth
particle surfaces were observed in the untreated specimen (Fig.
11a). For the treatment with cementation reagents only, some
calcite crystals were observed on the soil particles (Fig. 11b).

Fig. 12. SEM Images for the Sand Specimens: (a) Untreated, (b) Treated with Cementation Reagents Only, (c) Treated with B. megaterium
and Cementation Reagents
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KSCE Journal of Civil Engineering

Improvements in Engineering Properties of Soils through Microbial-Induced Calcite Precipitation

Abundance of calcite crystals were observed in the specimens


treated with B. megaterium and cementation reagents (Fig. 11c).
On closer observations, rod-shaped B. megaterium was found in
intimate contact with the calcite crystals.
Similar patterns were observed for the SEM images of sand
specimens (Fig. 12). Comparatively, the quantity of the precipitated
calcite crystal for the sand specimen treated with cementation
reagents only (Fig. 12b) was less than that observed in residual
soil specimen (Fig. 11b). This observation confirmed the results
from the shear strength and hydraulic conductivity tests that the
natural microorganisms only exist for an insignificant amount in
the sand specimens, and hence induced slight alterations to their
properties.
Furthermore, Energy Dispersion Spectroscopy (EDS) was
performed on the MICP-treated specimens using EDAX (AMETEK
Materials Analysis Division) to analyze the concentrations of
calcium (Ca), carbon (C), oxygen (O), and silica (Si) elements. It
is an analytical technique used for elemental analysis or chemical
characterization of samples. Fig. 13(a) illustrates the EDS
performed on a soil particle. The intensity of Si element was the
highest among all elements concerned since it is the major
element in soil. As for the Fig. 13(b), Ca and O recorded the

Fig. 13. EDS on (a) Soil Particles, (b) Crystal Calcite


Vol. 17, No. 4 / May 2013

highest intensity. An increased intensity was also encountered for


C. As these three elements form the main components of calcite
(CaCO3), the EDS was probably performed on a calcite crystal.

5. Discussion
Microbial induced calcite precipitation has been shown to be
an effective method to enhance the shear strength and reduce the
hydraulic conductivity of soil. The soil with enhanced strength
can contribute to a greater ground bearing capacity, while
reduced hydraulic conductivity can minimize settlement, shrinkswell tendency, seepage, and infiltration of rainfall into soils.
The experimental results indicated that MICP was more
effective in improving shear strength for residual soil (1.40-2.64
increment ratios) than for sand (1.14-1.25 increment ratios).
With respect to the reduction in hydraulic conductivity, the sand
specimen (0.09-0.15 reduction ratios) was found to be more
effective than the residual soil (0.26-0.46 reduction ratios). The
improvement in shear strength and diminution in hydraulic
conductivity of residual soil increased with increased density.
However, the sand specimens exhibited a reverse trend. These
different observations between residual soil and sand need
clarification through an insight into the behaviour of the B.
megaterium in soil.
According to Achal et al. (2009), the effectiveness of MICP
treatment on a soil specimen can be attributed to both the ability
of the microorganism to move freely throughout the pore space
and the sufficient particle-particle contacts per unit volume at
which cementation occurs. These two attributes, however, are
contradicting each other as the soil with large pore space tends to
have less particle-particle contacts per unit volume, and vice
versa. These conditions require a balance relationship between
the microorganism size and the pore structure characteristics,
namely the pore throats.
The relatively low improvements in shear strength of sand
compared to residual soil can be explained by the insufficient
concentrations of particle-particle contacts per unit volume. This
is because the sand specimen contains coarser granular particles.
The contacts between the coarse particles are lesser compared to
the residual soil specimen that consists of wide range of particle
sizes (ranging from smaller than 1 mm to 2 mm). The pores
between the coarse particles in residual soil are filled with the
smaller grains, thus results in greater particle-particle contacts.
The improved shear strength and reduced hydraulic conductivity
with increased density of residual soil can also be explained by
the particle-particle contacts. As the residual soil compacted to a
higher density, the particle-particle contacts increase. This
facilitates greater calcite bonding at particle-particle contacts.
Pore spaces also decrease with increased compaction. However,
the long treatment duration (2 days) and intermittent cementation
reagents injection method (flush the cementation reagents
through the sample every 6 hours at a moderately high velocity
of 1.7 105 m/s) employed in this study are believed to have
minimized the inverse impact of small pore throat on the MICP

726

Ng Wei Soon, Lee Min Lee, Tan Chew Khun, and Hii Siew Ling

treatment.
The sand specimen has a higher hydraulic conductivity
reduction ratio than the residual soil. This can be explained by
the greater porosity of sand. Greater porosity means more pore
space available for calcite deposition by B. megaterium, and
hence results in greater reduction in hydraulic conductivity.
As mentioned earlier, the pore throat size can also affect the
effectiveness of MICP. The improvement in shear strength and
reduction in hydraulic conductivity of the sand specimens
decreased with increased density. This is because denser sand
contributed to a smaller pore throat size. Consequently, the
movements of B. megaterium and reagent solutions within the
sand specimen were restrained, and hence retarded the MICP
process slightly. Although denser specimen may have greater
particle-particle contacts leading to enhanced improvement in
the soil properties, however, particle-particle contacts of sand
have not been improved by greater compaction due to lack of
finer particles that act as filler to the voids between the large
particles. The smaller pore throat in the sand specimen plays a
more dominant role in controlling the effectiveness of MICP.
The residual soil and sand specimens treated with cementation
reagents only exhibited slight increment in the shear strength and
diminution in the hydraulic conductivity. The results imply that
the amount of natural calcite forming microorganisms is insufficient
to trigger effective MICP. Comparatively, the residual soil
specimens show marginally greater improvements than the sand
specimens. This is because the residual soil specimen was taken
in-situ from a site which could be rich of natural microorganisms,
while the sand specimen was of typical concrete sand which was
left exposed under the extreme tropical climate. As the result, the
natural inhibited microorganisms in the sand specimens are
lesser than the residual soil specimens.

with the increased density. The results implied that the particle-particle contacts of sand have not been improved markedly by the higher degree of compaction.
4. The saturated hydraulic conductivities of the MICP-treated
residual soils exhibited reduction ratios of 0.26-0.45. The
reduction is less significant than the reduction ratios of the
sand specimens (0.09-0.15). This can be explained by the
greater porosity and pore spaces in sand that are available
for bioclogging.
5. The amounts of calcite precipitated in the treated residual
soil specimens ranged from 1.080% to 1.889%. The calcite
content increased with the increased soil density. Opposing
trend was observed for the sand specimens, whereby the
calcite content decreased with the increased sand density.
Nevertheless, the amounts of calcite in the treated sand
specimens (ranging from 2.661% to 6.102%) were generally higher than those of the residual soil specimens.
6. For both residual soil and sand specimens, treatment with
cementation reagents only exhibited slight alterations in
shear strength and hydraulic conductivity. The results indicated the presence of natural calcite forming microorganisms which existed in insignificant amount. The results
from SEM analysis confirmed this finding.
7. The effects of microorganism only (biomass) on the shear
strength and hydraulic conductivity of both residual soil and
sand specimens were negligible.

Acknowledgements
This project is funded by the Ministry of Higher Education,
Malaysia under Fundamental Research Grant Scheme (FRGS).
Besides, the authors acknowledge the useful comments from the
reviewers.

6. Conclusions
Twenty four configurations of experimental variables were
designed to investigate the effectiveness of MICP in improving
the shear strength and reducing the hydraulic conductivity of two
types of soil selected for the study. The following findings are
drawn from the study:
1. B. megaterium was demonstrated to enhance the shear strength
and reduce the hydraulic conductivity of both residual soil
and sand specimens. The improvement in the engineering
soil properties varied with soil densities, soil types, and
treatment conditions.
2. The MICP-treated residual soils exhibited significant increment ratios in shear strength, i.e., 1.41-2.64. The rate of
improvement increased with increased density. This can be
explained by high particle-particle contacts in residual soil
particles.
3. The MICP-treated sands improved in shear strength by
ratios of 1.14-1.25. The lower improvement compared to
residual soils can be attributed to the lesser contacts
between sand particles. The rate of improvement decreased

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