Lee / Jurnal Geoteknik HATTI IX(1)(2014) 30 - 39 | ISSN 0853 4810

Bio-Mediated Soil : A Sustainable Ground Improvement Technique

Lee Min Lee

Faculty of Engineering and Science, Univesiti Tunku Abdul Rahman, Kuala Lumpur, Malaysia
Email: mllee@utar.edu.my

ABSTRACT: Bio-mediated soil has recently emerged as a new and sustainable soil improvement technique. This
paper presents the results of investigation on the performances of bio-mediated soil improvement on sand and a
typical tropical residual soil (sandy silt). A species of Bacillus group, B. megaterium was used to trigger calcite
precipitation. The treatments were performed at various soil densities (85%, 90% and 95% maximum density), and
treatment conditions (untreated, treated with cementation reagents only, and treated with B. megaterium and
cementation reagents). Results showed that the MICP treatment which was often attempted on sand was equally
robust in tropically residual soil. The shear strength of MICP-treated residual soil was improved by 40 - 164%
compared to its untreated counterpart. The saturated hydraulic conductivity of the MICP-treated residual soil was
reduced by 54 - 74%. The effectiveness of the bio cementation and bio clogging in residual soil was mainly
governed by the particle-particle contacts, porosity and pore spaces between soil particles. Future research may
focus on in-situ applications of bio-mediated soil improvement and its effectiveness in fine-grained soils.

Keywords: bio-mediated soil improvement, microbially induced calcite precipitation, biocementation, bioclogging,
residual soil

ABSTRAK: Tanah bio - mediasi muncul sebagai teknik perbaikan tanah baru dan berkelanjutan. Makalah ini
menyajikan hasil penelitian mengenai kinerja perbaikan tanah bio - mediasi terhadap pasir dan tanah residual tropis
(sandy silt). Satu spesies dari kelompok Bacillus, B. megaterium digunakan untuk memicu produksi kalsit.
Perlakuan dilakukan pada berbagai kepadatan tanah (kepadatan maksimum 85 % , 90 % dan 95 % ), dan kondisi
perawatan ( tanpa campuran, dengan capuran reagen sementasi saja, dan dengan campuran B. megaterium dan
reagen sementasi ). Hasil penelitian menunjukkan bahwa perlakuan MICP yang sering dicoba pada pasir sama-sama
kuat di tanah tropis residual. Kekuatan geser tanah residual yang dicampur MICP meningkat 40-164 %
dibandingkan dengan tanah yang tidak dicampur . Konduktivitas hidrolik jenuh tanah residual yang dicampur MICP
berkurang sebesar 54-74 %. Efektivitas biocementation dan bioclogging di tanah residual terutama dikontrol oleh
kontak partikel - partikel, porositas dan ruang pori antara partikel tanah. Penelitian lanjutan dapat difokuskan pada
aplikasi metode perbaikan tanah bio - mediasi dan efektivitasnya dalam tanah butir halus .

Kata Kunci: perbaikan tanah dengan bio-mediasi, produksi kalsit yang dipicu oleh biomedia, biocementation,
bioclogging, tanah residual

1. INTRODUCTION
Ground improvement, in the context of geotechnical engineering, refers to enhancement of engineering
properties of soil, e.g. shear strength, hydraulic conductivity, compressibility, stiffness, etc. to
accommodate requirements of a specific construction. Common ground improvement techniques used in
current practice include mechanical compaction, dewatering, soil reinforcement and chemical grouting.
Selection of ground improvement technique lies on several factors such as soil type, thickness of soft soil
deposit, construction activities involved, and targeted soil properties to be improved. Chemical grouting is
commonly used to stabilize expansive and sensitive soil near the ground surface. Recent studies, however,
show that most of the chemical grouting are toxic and hazardous, despite of their proven effectiveness in
practice (DeJong, et al. 2010).
Bio-mediated soil improvement is a relatively new and innovative technique of ground improvement.
The technique takes advantages of natural bio-activities, technically termed as microbially induced calcite
precipitation (MICP), to produce calcite in soil matrix. The calcite generated is responsible for improving
engineering properties of soil. The MICP process has also shown promising applications in other
construction materials, i.e. improvements of strength (Siddique et al., 2008; Raijiwara et al., 2009) and
durability (De Muynck et al., 2008; Achal et al., 2011) of concrete / mortar, and durability of brick (Sarda
et al., 2009).
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The chemical reactions involved in the process of MICP is expressed as follows:

CO (NH2)2 + 2H2O 2NH4+ + CO32-

(1)

First, urea (CO(NH2)2) and water (H2O) are decomposed into ammonium (NH4+) and carbonate (CO32-)
ions with the presence of urease enzyme. This process is scientifically known as urea hydrolysis. It is
important to supply urease positive type bacteria, i.e. genera Bacillus, Sporosarcina, Spoloactobacilus,
Clostridium and Desulfotomaculum (Kucharski et al., 2008) into soil to promote the production of urease
enzyme, and hence the urea hydrolysis process .

The release of ammonium (NH4+) in the Eq. [1] is essential for increasing the pH of soil as the subsequent
calcite precipitation process favors a slightly alkaline environment. The carbonate (CO32-) ions will react
with the calcium ion (Ca2+) from the supplied calcium chloride to form calcium carbonate or calcite
(CaCO3):

Ca2+ + CO32- CaCO3

(2)

The calcite (CaCO3) precipitated is responsible for improving inherent engineering properties of soil
through biocementation and bioclogging. Biocementation is defined as an improvement of soil strength
by production of particle-binding materials through microbial means, while bioclogging is a reduction of
hydraulic conductivity of soil or porous rock by pore-filling materials generated by microbial processes
(Ivanov and Chu, 2008).
Bio-mediated soil improvement has been widely recognized as a 'greener' and more sustainable ground
improvement technique than the conventional chemical grouting method.
The treatment process involves relatively straight forward bio-chemical processes as depicted in Eqs.
(1) and (2). It is apparent that the MICP process requires the supply of three main ingredients / materials
only, namely urea (CO(NH2)2), calcium chloride, and urease positive bacteria. None of these materials are
toxic or hazardous to human health and soil organisms.
Current research of bio-mediated soil improvement has focused primarily on fine sands (DeJong et al,
2006). Studies on other types of soil are still very limited. This paper aims to investigate and compare the
performances of bio-mediated soil improvement on sand and a typical tropical residual soil (sandy silt).
The effectiveness of the MICP treatment was evaluated by direct measurements of shear strength and
saturated permeability of the soil specimens.

2. MATERIALS AND METHODS

2.1. Laboratory Setup

Figure 1 shows a schematic diagram of the experimental setup for MICP treatment. The apparatus
comprised a steel mold, an air compressor, a pressure tank, and an effluent collector. Air-dried residual
soil was first mixed with a culture medium containing urease-producing microorganism. The soil
specimen was then compacted into the steel mold to the desired densities. The specimen mold was
connected to the pressure tank that filled with cementation reagent solutions. The pressure in the pressure
tank was regulated by the air compressor to fix the flow pressure into the specimen mold at 1.1 bar.

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Figure 1. Schematic diagram of the laboratory setup

2.2. Soil Specimens

Two types of soil specimens were employed in this study, namely tropical residual soil and sand. The
tropical residual soil was obtained from a site in Universiti Tunku Abdul Rahman, Kuala Lumpur campus
compound, while the sand specimen was of typical concrete sand. The particle size distributions of the
two soil specimens are presented in Figure. 2. 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 to three different densities, i.e. 85% of MDD, 90% of MDD, and 95% of MDD. For the sand
specimen, 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.

Figure 2. Particle size distributions of soil specimens

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2.3. Microorganism and Growth Conditions

The urease-producing microorganism used in the present study was Bacillus megaterium (ATCC 14581).
The B. megaterium was cultivated at pH 7 under aerobic batch conditions in a sterile culture medium of 5
g/l peptone, 5 g/l sodium chloride, 2 g/l yeast extract, and 1 g/l beef extract. Incubation was performed in
a shaking incubator at 200 rpm and constant temperature of 37C. It was grown to early stationary phase
before harvesting at a concentration of approximately 1108 cfu/ml.

2.4. Cementation Reagent Solutions

The cementation reagent for the MICP treatment consisted of 0.5 M of urea and calcium chloride, and 3
g/l of nutrient broth supplement.

2.5. Soil Engineering Properties Tests

For the residual soil specimens, the shear strength was determined by performing unconfined
compression test on 50 mm diameter saturated specimens. The hydraulic conductivity was
determined by falling head permeability test. For the sand specimens, the shear strength was
determined by standard direct shear box test on 60 mm 60 mm dry specimens while the
hydraulic conductivity was determined by constant head permeability test.

2.6. Treatment Variables

The experimental design focused primarily 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
cementation reagents only, and treated with B. megaterium and cementation reagents) on the shear
strength and hydraulic conductivity of soils. 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.

3. RESULTS AND DISCUSSIONS

3.1. Shear Strength of Residual Soil

Figure. 3 shows the photographs of an untreated and a MICP-treated residual soil samples, while the
shear strength results are summarized in Fig. 4. The shear strength of the MICP-treated residual soils was
improved for all densities (85%, 90%, and 95% of maximum density). The shear strength improvement
increased with increased density, i.e. improved by 40%, 160% and 164% for specimens of 85%, 90%, and
95% MDD, respectively
The specimens treated with cementation reagents only also exhibited increased shear strength. The
undrained shear strength parameter improved by 11%, 26% and 33% for the specimens of 85%, 90%, and
95% MDD, respectively. The results implied that MICP was triggered by the microorganisms inhibiting
naturally in the soil deposits. The improvements (11-33%), however, were significantly lower than the
specimens treated with B. megaterium and cementation reagents (40-164%). 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.

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Figure 3. Photographs of untreated (left) and MICP-treated residual soil samples (right)

Figure 4. Shear strength improvements of residual soil specimens

3.2. Hydraulic Conductivity of Residual Soil

Figure 5 shows the results of hydraulic conductivity for the residual soil specimens. The saturated
hydraulic conductivity of MICP-treated soil was markedly reduced for all densities. The greatest
reduction in hydraulic conductivity occurred in the densest specimen (95% MDD) where the hydraulic
conductivity of the MICP-treated specimen was reduced by 74% compared to its untreated counterpart.
The reductions for 90% and 85% MDD specimens were 61% and 54%, respectively. In summary, the
reduction in hydraulic conductivity of soil was mainly inflicted by calcite and the effect was proportional
to the soil density. As the residual soil compacted to a higher density, the particle-particle contacts
increase. This facilitates greater calcite formed at particle-particle contacts. The formation of calcite
clogged most of the pores and reduced the saturated hydraulic conductivity of the residual soil effectively.

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Figure 5. Hydraulic conductivity reductions of residual soil specimens

3.3. Shear Strength of Sand

Figure 6 shows how the loose sand was transformed into a hardened cube upon MICP treatment. The
quantitative results of the sand effective internal friction angle are summarized in Fig. 7. The effective
internal friction angles of the untreated sand specimens were between 39.9o and 48.8o. The internal
friction angles of the MICP-treated sand specimens were generally higher than the untreated specimens.
The 85% max specimen had the greatest improvement (increased by 25%), followed by 90% max
specimen (increased by 17%) and 95% max specimen (increased by 14%). The shear strength
improvement decreased with increased density. The trend was opposite to the results observed in the
residual soil specimens. Furthermore, the sand specimens (improved by 14-25%) exhibited significantly
lower improvement than the residual soil specimens (improved by 40-164%). This phenomenon can be
explained by 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 a wide range of particle sizes (ranging from
smaller than 1 m to 2 mm). The pores between the coarse particles in residual soil are filled with the
smaller grains, and hence results in greater particle-particle contacts.
For the specimens treated with cementation reagents only, the shear strength improvement was
markedly lower (6-15%) compared to the specimens treated with B. megaterium and cementation
reagents (14-25%). The results implied that the amount of naturally inhibiting calcite forming
microorganisms in the sand specimens was negligible.

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MICP-treated sand
Loose sand

Figure 6. Photographs of untreated (left) and MICP-treated sand samples (right)

Figure 7. Shear strength improvements of sand specimens

3.4. Hydraulic Conductivity of Sand

The saturated hydraulic conductivities (ksat) of the sand specimens are presented in Fig. 8. Similar to the
trend observed in shear strength of sand specimens, the reduction in hydraulic conductivity becomes less
effective with the 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 absence of finer particles

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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 greatest reduction in hydraulic conductivity occurred in the 85% max specimen in which the
hydraulic conductivity decreased by approximately one order of magnitude from 3.5 10-3 m/s to 3.2
10-4 m/s (reduced by 91%). As the density of the specimen increased, the reductions of hydraulic
conductivity were marginally lesser as observed in 90% max
specimens (reduced by 85%).

3.5. Scanning Electron Microscopy Analysis

Scanning electron microscope (SEM) imagery was performed on selected specimens to visualize calcite
precipitations in the soil specimens. Fig. 9 compares the SEM images of an untreated and a MICP-treated
residual soil specimens. The untreated specimen has a considerably clean and smooth particle surface
(Figure. 9a). For the MICP-treated specimen, abundance of calcite crystals were clearly observed. Some
of the calcite crystals were precipitated between particle-particle contacts forming a strong calcite bond
between the soil particles (Figure. 9b).

Figure 8. Hydraulic conductivity reductions of sand specimens

(a)

(b)

Figure 9. SEM imagery of (a). Untreated specimen. (b). MICP-treated specimen

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4. DEVELOPMENT OF RESEARCH IN BIO-MEDIATED SOIL

Despite being a relatively young technology, many studies on bio-mediated soil improvement have been
reported over the past decade. In fact, the ability of MICP has been identified since early 2000, or earlier
(Bachmeier et al., 2002; Stocks-Fischer et al., 1999). DeJong et al. (2006) was among the pioneers who
attempted the MICP in geotechnical applications. Thereafter, many groups of researchers around the
world including Netherlands, United States, United Kingdom, India, China, and Australia have carried
out extensive research in this field. In recent years, bio-mediated soil improvement has become one of the
hottest and fastest growing research topics among geotechnical communities.
Ivanov and Chu (2008) presented a detailed review on the applications of MICP for soil improvement.
At present, promising applications focus primarily on biocementation, bioclogging and biogas. A
comprehensive review on the state of art research of bio-mediated soil has also been reported by a group
of experts in the area (DeJong et al., 2013). These reviews are found extremely useful for new researchers
who are interested to venture into this exciting soil improvement technique.
Most studies of MICP improvement have been conducted on a laboratory scale (Dejong, et al. 2006;
Dejong, et al. 2010; Whiffin, et al. 2007). Van Paassen (2011) provided an overview of the latest research
development in Netherlands, using scale-up laboratory tests and field-scale experiments. The MICP
technique has been applied successfully in field to strengthen the wall of borehole to avoid soil collapse
during drilling process. Future researches may look into the viability of the technique in improving
bearing capacity of in-situ soil. The performance of the bio-mediated soil improvement in fine-grained
soil also warrants an insightful research to broaden the applications of this relatively new soil
improvement technique in solving real-life geotechnical problems.

5. CONCLUSIONS
A series of laboratory tests were performed to investigate and compare the effectiveness of MICP in
improving shear strength and reducing hydraulic conductivity of sand and tropical residual soil.
Following findings are drawn from the study:

i) MICP was demonstrated to enhance the shear strength and reduce the hydraulic conductivity of
both residual soil and sand specimens. The improvement in the soil engineering properties varied
with soil densities, soil types, and treatment conditions.
ii) The MICP-treated residual soils exhibited significant increments in shear strength, i.e. improved by
40-164%. The rate of improvement increased with increased density. This can be explained by high
particle-particle contacts in residual soil particles.
iii) The MICP-treated sands improved in shear strength by 14-25%. The lower improvement compared
to residual soils can be attributed to the lesser contacts between sand particles.
iv) The saturated hydraulic conductivities of the MICP-treated residual soils were reduced by 54-74%.
These reductions were less significant than those of sand specimens (85-91%). This can be
explained by the greater porosity and pore spaces in sand that are available for bioclogging.
v) Future research in bio-mediated soil may focus on its in-situ applications and effectiveness of the
technique in improving fine-grained soils.

ACKNOWLEDGEMENTS
This project is funded by the Ministry of Higher Education, Malaysia under Fundamental Research Grant
Scheme (FRGS). Besides, the author would like to acknowledge the useful comments from the reviewer

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