Chowdariguda, Korremula (V), Ghatkesar (M), Medchal-Malkajgiri Dist.

, Ph: +918415255777
Fax:08415-25 5666, Telangana State-500088, Email: admin@nnrg.edu.in

SCHOOL OF ENGINEERING
DEPARTMENT OF CIVIL ENGINEERING

“Soil Stabilization by Bacterial Cementation”

submitted in partial fulfilment of the requirement for the award of the
degree of
BACHELOR OF TECHNOLOGY
in
CIVIL ENGINEERING

G SANNITH REDDY (H. No. 187Z1A0114)

K SHIVA KUMAR (H. No. 197Z5A0110)

V SRIJA (H. No. 197Z5A0120)

V KRISHNA SAI (H. No. 197Z5A0122)

Guide

Mr. M KARTHIK KUMAR R

Assistant Professor
 DECLARATION

We hereby declare that the Major Project entitled “Soil Stabilization by Bacterial Cementation”

presented in this report in partial fulfilment for the award of the degree of Bachelor of Technology

during the academic year 2021-22 in the Department of Civil Engineering, Nalla Narasimha Reddy

Education Society’s Group of Institutions affiliated to Jawaharlal Nehru Technological University,

Hyderabad.

We solemnly declare that to the best of my knowledge; no part of this report has been

submitted here or elsewhere in a previous application for the award of a degree. All sources of

knowledge used have been duly acknowledged.

PROJECT MEMBERS

G SANNITH REDDY (H. No. 187Z1A0114)

K SHIVA KUMAR (H. No. 197Z5A0110)

V SRIJA (H. No. 197Z5A0120)

V KRISHNA SAI (H. No. 197Z5A0122)

 Chowdariguda, Korremula (V), Ghatkesar (M), Medchal-Malkajgiri Dist., Ph: +918415255777
Fax:08415-25 5666, Telangana State-500088, Email: admin@nnrg.edu.in

SCHOOL OF ENGINEERING
DEPARTMENT OF CIVIL ENGINEERING

CERTIFICATE
This is to certify that the Major Project entitled

“Soil Stabilization by Bacterial Cementation”

carried out by
G SANNITH REDDY (H. No. 187Z1A0114)

K SHIVA KUMAR (H. No. 197Z5A0110)

V SRIJA (H. No. 197Z5A0120)

V KRISHNA SAI (H. No. 197Z5A0122)

have submitted and approved as per requirements and regulations of Governing Body for the award ofthe
degree of Bachelor of Technology in Civil Engineering affiliated to Jawaharlal Nehru Technological
University, Hyderabad, Telangana, India.

Mr. M. Karthik Kumar Date

(Internal Guide)

Mr. Y. Srinivas Date

(HOD-CE)

EXTERNAL EXAMINER Date

 ACKNOWLEDGEMENT

We wish to express our sincere thanks to Dr. C.V. Krishna Reddy, Director and
Dr. G. Janardhan Raju, Dean - School of Engineering, NNRESGI for providing all the necessary
facilities and their support.

We place on record, our sincere thanks to Mr. Y. Srinivas, Professor and HoD of Civil
Engineering Department NNRESGI for his whole-hearted cooperation, providing excellent lab
facilities with constant encouragement and unfailing inspiration.

We are highly grateful to Mr. M Karthik Kumar Reddy, Assistant Professor, Department of Civil
Engineering for extending his support and assisting us throughout the major project work. We are
immensely thankful to our guide for guiding us from the beginning of the project.

We whole heartedly thank Dr. C. Krishna Mohan, Dean- School of Pharmacy, NNRESGI and Mrs.
K. Prashanthi, Associate Professor, School of Pharmacy for their cooperation in providing lab
facilities and guiding us as a part of the project work.

We take this opportunity to record our sincere thanks to all the faculty of the Department of Civil
Engineering for giving timely suggestions during the progress of the mini project work.

Finally, we would like to thank our parents who have always encouraged us to do our best.

PROJECT MEMBERS

G SANNITH REDDY (H. No. 187Z1A0114)

K SHIVA KUMAR (H. No. 197Z5A0110)

V SRIJA (H. No. 197Z5A0120)

V KRISHNA SAI (H. No. 197Z5A0122)

 CONTENTS
Chapter No Description Page No.
Abstract i
List of Figures ii
List of tables iii-iv
List Graphs v
Symbols/Notations vi
1 Introduction 01-09
1.1 Expansive soils 02-03
1.2 Occurrence Expansive soils 04
1.3 Nature of expansive soil 04
1.4 Clay mineralogy 04-07
1.4.1 Kaolinite group 05
1.4.2 montmorillonite Group 06
1.4.3 Illite group 07
1.5 Identification &classification of
expansive soils 08
1.2 MICP Application 09

2 Literature Review 10-12

3 Materials & Methodology 13-40
3.1 Expansive soils 13
3.2 Bacteria 14
3.2.1 Bacillus subtills 14
3.3 Methodology of the Project 14-19
3.4 Methodology Adopted 19-40
3.4.1 Grain size distribution/sieve analysis 20-21
3.4.2 Change in plasticity index of soil -MICP 21-22
3.4.3 Standard Proctor Test 23-26
3.4.4 California Bearing Ratio 26-33
3.4.5 Unconfined Compressive Strength 33-40
Chapter No Description Page No.

4 Results 41

5 Conclusion 42
References 43-44
 ABSTRACT

In India nearly 51.8 million hectares of land area are covered with Expansive soil
(mainly Black Cottonsoil). The property of these expansive soils, in general, is that
they are very hard when in dry state, but they lose all of their strength when in wet
state. In light of this property of expansive soils, these soils pose problems worldwide
that serve as challenge to overcome for the Geotechnical engineers. One ofthe most
important aspects for construction purposes is soil stabilization, which is used widely
in foundation and road pavement constructions this is because such a stabilization
regime improves engineering properties of the soil, such as volume stability, strength
and durability. In this process, removal or replacing of the problematic soil is done
replacement is done by a better-quality material, or the soil is treated with an additive.
In the present study, using MICP(Microbiological Induced Calcite Precipitation).
stabilization of black cotton soil obtained from Huzurabad is attempted. With various
proportions of this additive i.e 5%, 10%, 15%, 20% & 25%, expansive soils is
stabilized. plasticityindex (P.I.) of clay-MICP mixes show a decrease in value with
increasing MICP content. In conclusion, addition of MICP results in decreases the
plasticity of the expansive soil, and increase in workability by changing its grain size
and colloidal reaction. the UCS & CBR values of clay with MICP mixeswere observed.
Analysis of the formerly found result exposes the potential of MICP as an additive
that could be used for improving the engineering properties of expansive soils.
Keywords : Soil stabilization, Black cotton soil,CBR,UCS.

i
 LIST OF FIGURES

Figure No Description Page No.

1.1 Major Soil Types in India 3

1.2 Atomic Structure of Kaolinite 5

1.3 Atomic Structure of Montmorillonite 6

1.4 Atomic Structure of Illite 7

3.1 Chemicals used for development of 16

bacteria
3.2 Adding of chemicals in conical flask 16

3.3 Sterilization of petri dishes & Conical 17

flask
3.4 Transformation of bacteria into the petri
dishes for development culture in UV 17
chamber
3.5 UV Chamber 18

3.6 Growth of the Cultured bacteria 18

3.7 Grain Size Distribution 20

3.8 Liquid Limit 21

3.9 Plastic Limit 21

3.10 Compacting the soil for tests 23

3.11 Soil Sample 23

3.12 Soil Compaction 26

3.13 Sample mould 26

3.14 UCS testing Machine 33

ii
 LIST OF TABLES

Table No. Description Page No.

1.1 Swelling potential vs Plasticity index 08

3.1 Sieve Analysis 20

3.2 Variation of plasticity index with Bacteria

22
in expansive soil.
[

3.3 Standard proctor test for expansive soil

23
only

3.4 Standard proctor test for expansive soil +

23
5% MICP by soil weight

3.5 Standard proctor test for expansive soil +

24
10% MICP by soil weight

3.6 Standard proctor test for expansive soil +

24
15% MICP by soil weight

3.7 Standard proctor test for expansive soil +

24
20% MICP by soil weight

3.8 Standard proctor test for expansive soil +

25
25% MICP by soil weight

3.9 CBR test for expansive soil only 27

[

3.10 CBR test for expansive soil + 5% MICP

28
by soil weight

3.11 CBR test for expansive soil + 10% MICP

29
by soil weight
[

3.12 CBR test for expansive soil + 15% MICP

30
by soil weight

iii
Table No. Description Page No.

3.14 CBR test for expansive soil + 20% MICP 31

by soil weight

3.15 CBR test for expansive soil + 25% MICP 32

by soil weight

3.16 UCS test for expansive soil + 5% MICP 35

by soil weight

3.17 UCS test for expansive soil + 10% MICP 36

by soil weight

3.18 UCS test for expansive soil + 15% MICP 37

by soil weight

3.19 UCS test for expansive soil + 20% MICP 38

by soil weight

3.20 UCS test for expansive soil + 25% MICP 39

by soil weight

5.1 Comparison of Index and Engineering 41

properties of MICP is mixed with soils

iv
 LIST OF GRAPHS

Graph no. Description Page No.

1 Sieve Analysis 21

2 Variation of plasticity index with MICP 22

3 Variation of MDD with MICP 25

4 Variation of OMC with MICP 26

5 Variation of CBR values with MICP by

soil weight 33

6 UCS test results 40

7 variation of UCS with MICP 40

v
 SYMBOLS/NOTATIONS

G Specific Gravity
% Percentage
OMC Optimum moisture content
MDD Maximum dry density
γd Dry density
γb Bulk Density
K Co-efficient of permeability
C Cohesion
 Angle of shearing resistance
SP Standard Publications
IS Indian Standard
BIS Bureau of Indian Standard
Wp Liquid Limit

Ws Plastic Limit

γdmax Maximum Dry Density

DST Direct shear test
PRR Proving ring reading
σ Normal Stress
σ Shear Stress
CBR California bearing ratio
µ Microns
MICP Microbiological Induced Calcite/Carbonate Precipitation
BCS Black cotton Soil

vi
 CHAPTER 1

INTRODUCTION
The need for stabilizing soils becomes necessary mainly because of two reasons: i) weak or
inconsistent soil properties and ii) need for urbanization especially in areas with problematic
soils such as expansive or high plasticity clays. These highly plastic soils cause heaving on the
ground surface (volume change) with change in moisture content. The change in moisture
content could be due to seasonal or climatic variations and evapotranspiration of vegetation.
Structures built on expansive soils tend to undergo moderate to severe cracking problems. In
particular, lightly loaded structures such as one or two story residential and industrial structures
and pavements often experience severe damage associated with substantive repair and
mitigation costs. Snethen, Townsend, Johnson, Patrick & Vedras (1975) in their report stated
that expansive soils are so widely distributed in United States that altering the highway routes
to avoid the expansive soils was virtually impossible. It is believed that the demand for new
and sustainable soil stabilization techniques, continues to grow with more than 40,000 soil
stabilization projects being carried out worldwide with total costs exceeding US$ 6 billion/year.
The artificial cementation of soil particles due to soil stabilization is often achieved through the
use of chemical stabilizers via shallow/deep mixing or injecting chemical grouts that can
permeate through soils ,Ismail, Joer, Sim & Randolph.

Physical properties of soil can be modified by the use of mechanical compaction and or
compaction grouting while chemical properties of soil can be modified by the use of chemical
stabilizers such as Portland cement, lime and fly ash. Mechanical compaction is recommended
for sandy soils and is effective or economical to a depth less than 10 m. Chemical stabilization
is typically recommended for expansive soils. Environmentally safe techniques such as
pre-wetting and moisture barriers are only possible for small confined spaces, and are not
suitable for larger construction projects such as highways and railways which spread for miles.
As mentioned above, artificial cementation techniques are not always feasible and
environmentally friendly. However, reduction in the use of artificial cementation techniques
can be practiced by substituting with environmentally friendly techniques or materials.
One such method of soil stabilization technique is, Microbial Induced Calcite Precipitation
(MICP). This technique employs microbes as a primary factor for stabilization.
Successful implementation of MICP will have its application in a wide variety of civil
engineering fields such as stability of retaining walls, embankments and dams, controlling soil
1
erosion, stabilizing cohesionless soils to facilitate the stability of underground constructions,
increasing bearing capacity of shallow and piled foundation and reducing the liquefaction
potential of soils.

1.1 EXPANSIVE SOILS

Expansive soils, which are also called as swell-shrink soil, have the tendency to shrink and
swell with variation in moisture content. As a result of this variation in the soil, significant
distress occurs in the soil, which is subsequently followed by damage to the overlying
structures. During periods of greater moisture, like monsoons, these soils imbibe the water, and
swell; subsequently, they become soft and their water holding capacity diminishes. As opposed
to this, in drier seasons, like summers, these soils lose the moisture held in them due to
evaporation, resulting in their becoming harder. Generally found in semi-arid and arid regions
of the globe, these types of soils are regarded as potential natural hazard – if not treated, these
can cause extensive damage to the structures built upon them, as well causing loss in human
life. Soils whose composition includes presence of montmorillonite, in general, display these
kinds of properties. Tallied in billions of dollars annually worldwide, these soils have caused
extensive damage to civil engineering structures. Also called as Black Cotton soils or Regur
soils, expansive soils in the Indian subcontinent are mainly found over the Deccan trap, which
includes Maharashtra, Andhra Pradesh, Gujarat, Madhya Pradesh, and some scattered places
in Odisha. These soils are also found in the river valley of Narmada, Tape, Godavari and
Krishna. The depth of black cotton soil is very large in the upper parts of Godavari and Krishna,
and the north-western part of Deccan Plateau. Basically, after the chemical decomposition of
rocks such as basalt by various decomposing agents, these are the residual soils left behind at
the place of such an event. Cooling of volcanic eruption (lava) and weathering another kind of
rock – igneous rocks – are also processes of formation of these type of soils. Rich in lime,
alumina, magnesia, and iron, these soils lack in nitrogen, phosphorus and organic content.
Consisting of high percentage of clay sized particles, the colour of this soil varies from black
to chestnut brown. 20% of the total land area, on an average, of this country is roofed by
expansive soils. These soils are suitable for dry farming and for the growth of crops like cotton,
rice, jowar, wheat, cereal, tobacco, sugarcane, oilseeds, citrus fruits and vegetables; the reason
behind it is owed to the moisture retentive capacity of expansive soils, which is high In the
semi-arid regions, just in the last couple of decades, damages due to the swelling- shrinking
action of expansive soils have been observed prominently in form of cracking and break-up of

2
roadways, channel and reservoir linings, pavements, building foundations, water lines,
irrigation systems, sewer lines, and slab-on-grade members.

Fig 1.1 Major soil types in India

3
1.2 Occurrence of expansive soil
Clay mineral is the key element which divulges the swelling characteristics to any ordinary
non- swelling/non-shrinking soil. Montmorillonite, out of several types of clay minerals has
the maximum amount of swelling potential. In-situ formation of chief clay minerals occurs
under alkaline conditions, or sub-aqueous decomposition of blast rocks can be seen the origin
of such soil – expansive soil. This type of soil can also be formed due to weathering under
alkaline environments, and under adequate supply of magnesium or ferric or ferrous oxides.
Given there’s a good availability of alumina and silica, the formation of Montmorillonite is
favored.
1.3 Nature of expansive soil
Swelling in clays can be sub-categorized into two distinctive types, namely:
1. Elastic rebound in the compressed soil mass due to reduction in compressive force.
2. Imbibing of water resulting in expansion of water-sensitive clays.
Swelling clays are the clays that exhibit latter type of swelling, where the clay minerals
with largely inflating lattice are present. One of the fundamental characteristics of
clayey soil is that they display little cohesion and strength when wet, but they become
hard when devoid of water. However, all of them do not swell due to wetting action.
Decrease in ultimate bearing capacity at saturation, and large differential settlement due
to this occurs. Thus, clayey soils exhibit foundation problems.
1.4 Clay Mineralogy
On the basis of their crystalline arrangement, clay minerals can be categorized into
three general groups,namely:

• Kaolinite group
• Montmorillonite group
• Illite group

4
1.4.1 Kaolinite group

A clay mineral which has a chemical composition 𝐴𝑙2𝑆𝑖205(𝑂𝐻)4 is called Kaolinite. This type
of clay mineral has a layered silicate, with linkage to one octahedral sheet of alumina through
oxygen atoms. China clay or Kaolin is the name given to rocks that are rich in this mineral. A
thickness of 7Å is exhibited by the stacked layers of kaolinite; as a result of this, kaolin group
of minerals are seen to be the most stable, which is also because of the fact that water cannot
enter between the sheets to inflate that unit cell.

Figure 1.2 Atomic structure of Kaolinite

5
1.4.2 Montmorillonite group

Two silica tetrahedral sheets combined with a central alumina octahedral sheet comprise the
structural arrangement of Montmorillonite. The bond between crystal links is weak here. Thus,
the soil containing higher percentage of Montmorillonite minerals demonstrate high shrinkage
and swelling characteristics, depending on the nature of exchangeable cation present. The
common layer of a Montmorillonite unit is formed by one of the hydroxyl layers of the
octahedral sheet and the tips of the tetrahedrons from each silica sheet. Atoms which are
common to both silica and gibbsite layers never participate in the process of swelling. During
weak bond between the crystal forms, water can penetrate, breaking the structures to 10Å
structural units.

Figure 1.3 Atomic structure of Montmorillonite

6
1.4.3 Illite group

As far as structural arrangement is concerned, Illite minerals fall between Montmorillonite and
Kaolinite group. As in case of Montmorillonite unit structure, two silica tetrahedral sheets
combined with a central alumina octahedral sheet comprise the structural arrangement of Illite.
The spacing between the elementary silica-gibbsite-silica sheets depend largely upon the
availability of water to occupy the space. Owing to this reason, Montmorillonite is believed to
have an expanding lattice. However, in presence of excess water, Illite can split up into
individual layers of 10Å thick.

Figure 1.4 Atomic structure of Illite

7
 1.5 Identification and classification of expansive soils

Some laboratory tests are available for the identification purposes of swelling soils.
By differential thermal analysis, microscopic examination, and X-ray diffraction. The presence
ofMontmorillonite in clay minerals allows the judgement of the expansiveness of the soil. This
aspect is however very technical in nature. A simple aspect, as opposed to the aforementioned
methods, is the free-swell test, that’s done in the laboratory. This test is conducted by adding
10 gm of dry soil, passing through a 425 μ sieve into two separate 100 cc graduated jar – one
filled with water, and the other with kerosene. Swelling occurs in the jar containing water. The
swelled volume of the soil is then noted (after 24 hours period), and subsequently, the free swell
index values, in percentage, are calculated. IS: 2720-II was followed for free swell indextest.

Free swell value [𝐼𝑛]

(in %age) = (𝐹𝑖𝑛𝑎𝑙 𝑣𝑜𝑙𝑢𝑚𝑒−𝑖𝑛𝑖𝑡𝑖𝑎𝑙 𝑣𝑜𝑙𝑢𝑚𝑒) 𝑥 100
𝑖𝑛𝑖𝑡𝑖𝑎𝑙 𝑣𝑜𝑙𝑢𝑚𝑒
Good grade, high swelling, commercial Bentonite has been reported to have free swell values
varying from 1200% to 2000%. In general, the swelling potential of a soil is related to plasticity
index. With corresponding range of plasticity index, various degrees of swelling capacities are
as indicated through the following table:

Table 1.1 Swelling potential vs. Plasticity Index

Swelling potential Plasticity Index

Low 0-15
Medium 15-24
High 24-46
Very High >46

Several factors participate in deciding whether or not a soil with high swelling potential exhibit
swelling characteristics. One of these factors, that occupy greatest importance, is the difference
between soil moisture content at the time of construction, and final (equilibrium) moisture content
finally achieved under various conditions allied with the complicated structure. The soil has a high
swelling capacity if the equilibrium moisture content is higher than the soil moisture content. Large
swelling pressure may develop as a result of the upheaving of the soil or structure, causing swelling.

8
1.6 MICP APPLICATIONS

Microbes are often responsible for the chemical cementation of soils in nature due to the precipitation
of cementing materials into the voids of soils and rocks. Microbes are able to precipitate cementing
materials such as calcium, magnesium, iron, manganese and aluminium which are crystallized to form
carbonates, silicates, phosphates, sulphides and hydroxides. The prime role of microbes in the
precipitation of minerals is their ability to create an alkaline environment through various
physiological activities. Calcium carbonate (calcite) precipitation is observed to be a general mineral
precipitation process in the microbial world under ambient environment.

Soil stabilization via MICP is one of several applications of bio-remediated processes. MICP can be
used for the elimination of soluble calcium from wastewater generated by industries. The high
calcium concentration in water can clog pipes and malfunction reactors. concluded that soluble
calcium was precipitated in the form of calcite by the use of bacteria as an alternative for chemical
precipitation of calcite. Thus, making MICP as an alternative and environmentally friendly technique
for the removal of calcium from industrial waste water. Ramachandran, Ramakrishnan & Bang
studied the effect of MICP on the compressive strength of Portland Cement by mixing urease enzyme
producing bacteria, Sproutarian. Pasteurian with cement mortar. For this purpose, they prepared 5 cm
cube moulds containing cement and bacteria, and were cured for 28 days in urea/calcium solution.
The cube was tested for compressive strength. It was reported that the strength increased by 24%
compared to untreated cube. Urease producing bacteria have been used in the oil industry to reduce
the permeability of the surface and subsurface media thus reducing the flow of the fluid and enhancing
the recovery of oil from reservoirs and limiting the spread of the contaminants from a spill site. This
process is called mineral plugging. The increase in pH due to the formation of ammonia as a by-
product during the breakdown of urea in the presence of urease enzyme as a catalyst, this increase in
pH provides a favourable condition for the precipitation of calcite in the presence of calcium ions.

MICP technique is considered to be a better and more environmentally friendly alternative to the
conventional technologies. However, more investigations are needed to properly understand the
possibilities and limitations. Further, its application in effectively stabilizing expansive soils is still a
hypothesis and this research is an initial step in evaluating this hypothesis and understanding the
applicability of MICP technique to stabilize expansive soils.

9
 CHAPTER 2

LITERATURE REVIEW
Krishna Kumari (2021) The challenges to enhance the weak soil constantly lay a need for research
and exploration to flourish a new and advance method for soil stabilization. The feasible technique is
Bacterial cementation which has evolved recently and raised a sustainable technique for soil
improvement. To analyse the potency of MICP (Microbiologically induced calcium carbonate
precipitation) in developing shear strength and lowering the hydraulic conductivity of soil.
Laboratory has been done to certify the effect of Bio grout (Bacterial Cementation) process on the
strength of 2 types of sand (High Plastic clay and Low plastic clay). The species of Bacillus group,
Magisterium was utilized to activate the calcite precipitation. The strain of Bacillus group was utilized
to trigger and catalyse the calcite precipitation due to chemical reaction of calcium chloride and urea.
MICP utilizes bacteria to hydrolyse urea and provide carbonate ions and that reacts with the calcium
chloride solution which provides calcium carbonate (calcite) that binds and holds the soil particles
together leads to increase the strength of soils and raise stiffness. It was also found that the strength
developed with advancement in treatment duration. The outcome of this experiment was evidently
shows from the pictures of Scanning Electron Microscope (SEM).

Balu Soundarya, Ponnusamy, Kulanthaivel, Subramaniam, Nithipandian and Venkata

salapathi Soundaryan 2020 To gratify the escalating need for land, it is vital to improve the lands
with loose weak soils. These problematic loose soils can be optimized by nurturing the shear strength,
lessening the time and potential for settlement and also reducing the hydraulic conductivity of soil,
by using Microbial induced calcite precipitation (MICP) process. This process of MICP is carried out
by adding bacterial solution into soil specimen which is continued with inoculation of cementation
reagents having urea and one calcium salt (CaCl2) for enormous times. As a result, calcite precipitate
(CaCO3) is formed in the soil and stabilize the soil. In this study, a number of factors that are
responsible for better formation of calcite precipitate are examined. This process mainly depends
upon soil type, bacteria species, concentration of cementation solution, injection methods and also on
pH, temperature conditions and curing period. The aim of this paper is to review the main factors that
influence the MICP process, such as soil properties, bacterial species, nutrients, concentration of
cementation solution, pH, temperature and injection methods.

Jinhua Zhang, Xizhi Shi, Xin Chen, Xiaofeng Hue, and Zi Yu 2020 School of Resources and
Safety Engineering, Central South University (CSU), Based on recent literary sources, this survey

10
discusses the effects of main factors influencing the microbial-induced calcium carbonate
precipitation (MICP), including the bacterial species, bacterial concentration, temperature, and pH
value. While the MICP technology has been widely adopted to improve rock and soil characteristics,
it has excellent development prospects in many other fields. Positive breakthrough solutions in the
MICP technology are improving geotechnical and foundation sand properties, repairing cement-
based materials, using mineralized film mulching to protect cultural relics, enhancing properties of
tailings, desert control, and heavy metal environmental restoration, etc., are discussed. experimental
findings prove that MICP can improve the strength, stiffness, liquefaction resistance, erosion
resistance, and permeability of geotechnical materials and maintain the good permeability and
permeability of the soil and improve the growth environment of plants. It is an environment friendly
bioengineering technology. Because microbial mineralization involves a series of biochemical and
ionic chemical reactions, there are many reaction steps in the solidification process and the
solidification effect of MICP is restricted and affected by many factors. +e comprehensive analysis
and optimization strategy on MICP industrial implementation should account for micro- and macro-
scale effects: the type of bacteria, the concentration of bacteria and cementation solutions, ambient
temperature, pH value, and other factors directly affect the crystallization type, morphology, and size
of calcium carbonate from the microscopic standpoint, while the macro-scale factors control the rock
and soil mineralization. +e limitations and prospects of the MICP technology are outlined.

Yasin Dursun Sari 2014 Civil Engineering Department, Faculty of Engineering and Architecture,
Istanbul Gallisin University, Istanbul, Turkey 6 August 2014 the biologically induced cementation
(Bio Grout) method has been used in this study to improve engineering properties of soil. Laboratory
tests have been carried out to quantify the effect of Bio Grout treatment on the strength of two types
of sand by using Sproutarian Pasteurian bacteria. Bearing strength and other parameters, such as
colony forming units (number of bacteria), pH level, temperature, and amount of CaCO3, have also
been studied. Calcification (cementation) through precipitation of CaCO3 on the mineral surfaces has
been clearly observed from the images of a scanning electron microscope (SEM). Finally, the effect
of sand material on the effectiveness of bio cementation was investigated via employment of different
sand types.

Cheng, L. and Shahin, M. 2017 The use of microbiological processes to improve the mechanical
properties of soil has gained some attention. This paper explores an emerging and promising
biological soil stabilization technique, known as bio-cementation, using microbially induced calcite
precipitation (MICP). In this work, uniform silica sand was treated using bio-cementation as well as
ordinary Portland cement (OPC), and the results were compared in terms of the unconfined

11
compressive strength (UCS) and permeability. The results indicate that bio-cementation is an
effective soil stabilization technique in improving soil strength, with higher achieved UCS values and
retained permeability than those of OPC-treated soil, demonstrating a major advantage for bio-
cementation. The effectiveness of bio-cementation in harsh environment of extremely low and high
temperatures was also examined, investigating the potential use of this technique in broader
conditions in cold and arid regions. Furthermore, the performance of bio-cementation in marine
environment was evaluated, showing the possibility of utilizing seawater as a natural calcium source
to replace commercially available calcium chloride and demonstrating the feasibility of this technique
in marine applications.

Masharu Fukue, Shin-Ich Ono and Yoshio Sato 2011 Microbial precipitation of carbonate was
observed using high microbial urease activity, and it was found that the ratio of Mg/Ca asserted the
types of crystals produced. Without Mg2＋, calcite was produced using only CaCl2, while the
presence of Mg produced Mg-calcite, magnesite and/or possibly dolomite of round, spherical or bores
shapes, depending on reaction time, pH and Mg/Ca ratio. The carbonate produced contributed to the
development of cementation for sands. The presence of Mg showed a relatively strong cementation
of the carbonate.

12
 CHAPTER 3
MATERIALS & METHODOLOGY

3.1 Expansive soil

As a part of this investigation, the expansive black cotton soil was acquired from the Huzurabad,
Telangana. The black cotton soil thus obtained was carried to the laboratory in sacks. A small amount
of soil was taken, sieved through 4.75 mm sieve, weighed, and air-dried before weighing again to
determine the natural moisture content of the same.

3.2 Bacteria

Bacteria are microscopic organisms, single-celled prokaryotic creatures. Bacteria come in

different shapes and the sizes. Bacteria are ubiquitous in every habitat on Earth, growing in soil,
acidic hot springs, radioactive waste, water, and deep in the Earth’s crust, as well as in organic
matter and the live bodies of plants and animals. There are typically 40 million bacterial cells in
a gram of soil and a million bacterial cells in a milliliter of fresh water; in all, there are
approximately five nonillion (5×1030) bacteria on Earth (Whitman et al. 1998, Vol.95) forming
much of the world’s biomass.

Bacteria were first observed by Antoine van Leeuwenhoek in 1676, using a single-lens
microscope of his own design. He called them “animalcules” and published his observations in
a series of letters to the Royal Society. The name bacterium was introduced much later, by
Christian Gottfried Ehrenberg in 1838.

Louis Pasteur demonstrated in 1859 that the fermentation process is caused by the growth of
microorganisms. Along with his contemporary, Robert Koch, Pasteur was an early advocate of
the germ theory of disease.

There are broadly speaking two different types of cell wall in bacteria, called Gram- positive
and Gram-negative. The names originate from the reaction of cells to the Gram stain, a test long
employed for the classification of bacterial species.

In the laboratory, bacteria are usually grown using solid or liquid media. Solid growth media
such as agar plates are used to isolate pure cultures of a bacterial strain. However, liquid growth
media are used when measurement of growth or large volumes of cells are required. Growth in
stirred liquid media occurs as an even cell suspension, making the cultures easy to divide and
transfer, although isolating single bacteria from liquid media is difficult. The use of selective

13
media (media with specific nutrients added or deficient or with antibiotics added) can help
identify specific organisms.

Bacterial growth follows three phases. When a population of bacteria first enters a high- nutrient
environment that allows growth, the cells need to adapt to their new environment. The first phase
of growth is the lag phase, a period of slow growth when the cells are adapting to the high-
nutrient environment and preparing for fast growth.

The lag phase has high biosynthesis rates, as proteins necessary for rapid growth are produced.
The second phase of growth is the logarithmic phase (log phase), also knownas the exponential
phase. The log phase is marked by rapid exponential growth. The rate at which cells grow during
this phase is known as the growth rate, and the time taken for the cells to double is known as the
generation time. During log phase, nutrientsare 14metabolized at maximum speed until one of
the nutrients is depleted and starts limiting growth. The final phase of growth is the stationary
phase and is caused by depleted nutrients. The cells reduce their metabolic activity and consume
non-essentialcellular proteins.

3.2.1 Bacillus subtillus

Researchers with different bacteria proposed different bacterial cementation in soils. The various
bacteria used in the bacterial cementation are Bacillus pasteurii, Bacillus sphearicus, E. coli etc.
In the present study an attempt was made by using the bacteria Bacillus subtilis strain no. JC3.
The main advantage of embedding bacteria in the soils is that it can constantlyprecipitate calcite.
This phenomenon is called microbiologically induced calcite precipitation (MICP). Calcium
carbonate precipitation, a widespread phenomenon among bacteria, has been investigated due
to its wide range of scientific and technological implications. Bacillus subtilis is a laboratory
cultured soil bacterium andits effect on the strength and durability is studied here.

3.3 Methodology of the project

The present work is divided into two phases, they are

Phase I – Developing Culture of bacteria

Phase II– Conducting tests in Laboratory, i.e., Grain size distribution, Liquid limit, Plastic limit,
CBR, UCS, etc.

14
Phase I – Developing Culture of bacteria

Petri Dishes, Glass Tubes, Conical Flasks are sanitized using the Autoclave machine in the
laboratory. Raw Bacteria Bacillus Subtillus is brought from the market. Bacteria culture is
developed by taking a small amount of the raw bacteria under the UV radiation zone in the
pharmacy laboratory in our institution. Distilled Water and Food for bacteria is taken with
different chemicals mixed with the distilled water.
The chemicals which are used as food nutrient for the development of bacteria cultureis that:

In a 250ml of conical flask, the following are taken:

➢ Beef extract powder- 1%

➢ Peptone- 1%

➢ Sodium chloride- 0.5%

➢ Agar agar Powder – 2%

This mixture is stored in a 10ml glass tubes and raw bacteria of 1 ml is inserted into theglass
tubes. These tubes are done again sterilization in the Autoclave machine at a pressure of
50lb/inch2 by keeping the cotton plugs such that no liquid comes out it when high pressure is
applied in the machine.
Then, this liquid is applied on the petri dishes and waited until it becomes a solid (in the form of
a wax). Streaks are drawn on it so that to identify the development of cultureof the bacteria. These
petri dishes are placed in the BOD incubator for 48 hours and thebacteria culture is developed
after the removal of the petri dishes from the Incubator. The developed bacteria can be seen with
the naked eye which forms in a white color.

15
Fig 3.1 Chemicals used as Nutrients for development of bacteria culture

Fig 3.2 Adding the chemicals in the conical flask

16
 Fig 3.3 Sterilization of Petri Dishes and Conical Flask

The developed culture is taken with a small amount using stirrer and placed in the
required quantity of water that is used in the bacterial cementation production. A conical
flask is taken and distilled water poured into it and a small number of bacteria is
inoculated into the flask. This flask is again placed in the Autoclave machine for the
proper mixture of bacteria into the water.

Fig 3.4 Transformation of Bacteria into the petri dishes for developmentof culture in the
presence of ignition in UV Chamber

17
 Fig 3.5 UV Chamber

Fig 3.6 Growth of the Cultured Bacteria

18
 Phase II – Conducting tests in laboratory
i.e. Grain size distribution, Liquid limit, Plastic limit, CBR, UCS, etc.…

3.4 Methodology Adopted

To evaluate the effect of MICP as a stabilizing additive in expansive soils, series of tests, where the
content of MICP in the expansive soil was varied in values of 5% to 25% (multiples of 10) by weight
of the total quantity taken. The Indian Standard codes were followed during the conduction of the
following experiments:

o Liquid & Plastic limit test – IS 2720 (Part 5) – 1985

o Standard proctor test – IS 2720 (Part 7) – 1980

o California bearing ratio (CBR) test – IS 2720 (Part 16) – 1987

o Unconfined compressive strength (UCS) test – IS 2720 (Part 10) – 1991

B5: Bacterial cementation by 5% of replacement of water with bacteria

B10: Bacterial cementation by 10% of replacement of water with bacteria
B15: Bacterial cementation by 15% of replacement of water with bacteria
B20: Bacterial cementation by 20% of replacement of water with bacteria
B25: Bacterial cementation by 25% of replacement of water with bacteria

The above tests are done with proper attention and recorded the values. Compare the bacterial
Cementation recorded values with-without Bacterial cementation values and plotted the graph
between them.

19
 3.4.1 Grain size distribution

Fig 3.7 Grain Size distribution

Table 3.1 Sieve Analysis

Sieve size Weight Cumulative weight Cumulative %

% Finer
Retained retained retained
4.75 83 83 27.67 72.33
1.00 19 102 34 66
0.600 75 177 59 41
0.425 13 190 63.33 36.67
0.300 34 224 74.67 25.33
0.212 17 241 80.33 19.67
0.150 18 259 86.33 13.67
0.075 31 290 96.67 3.33
Pan 10 300 100 0

20
 70
Sieve Analysis
60

50

40
% of finer

30

20

10

0
0 0.2 0.4 0.6 0.8 1 1.2
Sieve size

Graph 1 Plot of grain size analysis

3.4.2 Liquid Limit & Plastic Limit

Fig 3.8 Liquid Limit Fig 3.9 Plastic Limit

21
Table 3.2 Variation of plasticity index with Bacteria in expansive soil.
Mixture Liquid limit Plastic limit Plasticity index
Only soil 62.3 34.6 27.7
Soil + 5% 59.4 33.2 26.2
Soil + 10% 52.6 30.2 22.4
Soil + 15% 49.9 29.5 20.4
Soil + 20% 46.7 28.4 18.3
Soil + 25% 43.2 26.3 16.9

variation of plasticity index with MICP

30

25

20
Plasticity Index

15

10

0
0 5 10 15 20 25 30
% of MICP

Graph 2 Plot variation of plasticity index with MICP

22
 3.4.3 Standard Proctor Compaction Test

Fig 3.10 Compacting the soil for tests Fig 3.11 Soil sample

Table 3.3 Standard proctor test for expansive soil only

Weight of soil Moist unit Dry unit

Volume of Moisture
in mould weight weight
Mould (c𝒎𝟑) content (%)
(g) (g/𝐜𝒎𝟑) (Kg/𝐜𝒎𝟑)
997.46 5560 5.56 17.76 1.32
997.46 5730 5.73 19.53 1.45
997.46 5860 5.86 22.65 1.52
997.46 5870 5.87 24.87 1.50
997.46 5820 5.82 27.92 1.42

Table 3.4 Standard proctor test for expansive soil + 5% MICP by soil weight

Weight of Moist unit Dry unit

Volume of Moisture
soil in mould weight weight
Mould (c𝒎𝟑) content (%)
(g) (g/𝐜𝒎𝟑) (g/𝐜𝒎𝟑)
997.46 5600 5.60 15.18 1.39
997.46 5760 5.76 19.09 1.48
997.46 5830 5.83 22.13 1.50
997.46 5800 5.80 27.96 1.41
997.46 5770 5.77 32.71 1.33

23
 Table 3.5 Standard proctor test for expansive soil + 10% MICP by soil weight

Weight of Moist unit Dry unit

Volume of Moisture
soil in mould weight weight
Mould (c𝒎𝟑) content (%)
(g) (g/𝐜𝒎𝟑) (g/𝐜𝒎𝟑)
997.46 5620 5.62 19.6 1.36
997.46 5720 5.72 20.95 1.42
997.46 5860 5.86 22.56 1.52
997.46 5870 5.87 25.21 1.50
997.46 5820 5.82 29.13 1.41

Table 3.6 Standard proctor test for expansive soil + 15% MICP by soil weight

Weight of soil Moist unit Dry unit

Volume of Moisture
in mould weight weight
Mould (c𝒎𝟑) content (%)
(g) (g/𝐜𝒎𝟑) (g/𝐜𝒎𝟑)
997.46 5640 5.64 15.12 1.43
997.46 5760 5.76 18.96 1.48
997.46 5860 5.86 21.27 1.53
997.46 5870 5.87 24.71 1.50
997.46 5820 5.82 28.13 1.42

Table 3.7 Standard proctor test for expansive soil +20% MICP by soil weight

Weight of Moist unit Dry unit

Volume of Moisture
soil in mould weight weight
Mould (c𝒎𝟑) content (%)
(g) (g/𝐜𝒎𝟑) (g/𝐜𝒎𝟑)
997.46 5590 5.59 16.22 1.37
997.46 5710 5.71 18.52 1.45
997.46 5820 5.82 23.57 1.47
997.46 5840 5.84 26.42 1.46
997.46 5180 5.80 31.81 1.36

24
Table 3.8 Standard proctor test for expansive soil + 25% MICP by soil weight

Weight of soil Moist unit Dry unit

Volume of Moisture
in mould weight weight
Mould (c𝒎𝟑) content (%)
(g) (g/𝐜𝒎𝟑) (g/𝐜𝒎𝟑)

997.46 5530 5.53 14.11 1.34

997.46 5610 5.61 17.23 1.37
997.46 5730 5.73 21.34 1.42
997.46 5780 5.78 25.76 1.42
997.46 5760 5.76 31.2 1.34

Variation of MDD with MICP

1.54

1.52

1.5

1.48
Yd

1.46

1.44

1.42

1.4
0 5 10 15 20 25 30
% of MICP

Yd

Graph 3 Plot variation of MDD with MICP

25
 Variation of OMC with MICP

25

20
Moisture content %

15

10

0
0 5 10 15 20 25 30
% of MICP by soil weight

Graph 4 Plot variation of OMC with MICP

3.4.4 California Bearing Ratio

Fig 3.12 Soil Compaction Fig 3.13 Sample Mould

26
Table 3.9 CBR test for expansive soil

Standard
load
Plunger Dial CBR CBR
Applied intensity
penetration gauge
Sl. No. stress (𝒌𝒈/ intensity
(𝒎𝒎) readings load (𝒌𝒈)
(𝒌𝒈/𝒄𝒎𝟐) 𝒄𝒎𝟐) (% 𝒂𝒈𝒆)

1 0.0 1 2.47 0.12

2 0.5 6 14.82 0.75
3 1.0 17 42.01 2.14
4 1.5 24 59.30 3.02
5 2.0 28 69.19 3.52
6 2.5 32 79.07 4.03 70 5.75
7 3.0 36 88.95 4.53
8 3.5 40 98.84 5.03
9 4.0 44 108.72 5.54
10 4.5 47 116.14 5.91
11 5.0 50 123.55 6.29 105 5.99
12 5.5 52 128.49 6.54
13 6.0 53 130.96 6.67
14 6.5 54 133.43 6.79
15 7.0 55 135.90 6.92
16 7.5 57 140.84 7.17 134 5.35
17 8.0 58 143.32 7.30
18 8.5 58 143.32 7.30

27
Table 3.10 CBR test for expansive soil + 5% MICP by soil weight

Standard
load
Plunger Dial CBR CBR
Applied intensity
penetration gauge
Sl. No. stress (𝒌𝒈/ intensity
(𝒎𝒎) readings load (𝒌𝒈)
(𝒌𝒈/𝒄𝒎𝟐) 𝒄𝒎𝟐) (% 𝒂𝒈𝒆)

1 0.0 4 9.88 0.49

2 0.5 10 24.71 1.23
3 1.0 28 69.19 3.45
4 1.5 44 108.72 5.43
5 2.0 52 128.49 6.42
6 2.5 59 145.79 7.28 70 10.40
7 3.0 66 163.08 8.14
8 3.5 72 177.91 8.88
9 4.0 78 192.74 9.63
10 4.5 83 205.09 10.24
11 5.0 90 222.39 11.11 105 10.58
12 5.5 95 234.74 11.72
13 6.0 98 242.16 12.09
14 6.5 100 247.10 12.34
15 7.0 101 249.57 12.46
16 7.5 102 252.04 12.59 134 9.39
17 8.0 102 252.04 12.59

28
Table 3.11 CBR test for expansive soil + 10 % MICP by soil weight

Standard
load
Plunger Dial CBR CBR
Applied intensity
penetration gauge
Sl. No. stress (𝒌𝒈/ intensity
(𝒎𝒎) readings load (𝒌𝒈)
(𝒌𝒈/𝒄𝒎𝟐) 𝒄𝒎𝟐) (% 𝒂𝒈𝒆)

1 0.0 8 19.77 1.01

2 0.5 25 61.77 3.15
3 1.0 42 103.78 5.29
4 1.5 60 148.26 7.55
5 2.0 73 180.38 9.19
6 2.5 83 205.09 10.45 70 14.93
7 3.0 96 237.21 12.08
8 3.5 110 271.81 13.85
9 4.0 122 301.46 15.36
10 4.5 133 328.64 16.74
11 5.0 139 343.47 17.50 105 16.67
12 5.5 146 360.76 18.38
13 6.0 153 378.06 19.26
14 6.5 157 387.94 19.76
15 7.0 159 392.89 20.02
16 7.5 161 397.83 20.27
17 8.0 163 402.77 20.52 134 15.31
18 8.5 164 405.24 20.65
19 9.0 165 407.71 20.77
20 9.5 165 407.71 20.77

29
Table 3.12 CBR test for expansive soil + 15% MICP by soil weight

Standard
load
Plunger Dial CBR CBR
Applied intensity
penetration gauge
Sl. No. stress (𝒌𝒈/ intensity
(𝒎𝒎) readings load (𝒌𝒈)
(𝒌𝒈/𝒄𝒎𝟐) 𝒄𝒎𝟐) (% 𝒂𝒈𝒆)

1 0.0 3 7.41 0.37

2 0.5 9 22.24 1.13
3 1.0 19 46.95 2.39
4 1.5 29 71.66 3.65
5 2.0 37 91.42 4.66
6 2.5 52 128.49 6.54 70 9.35
7 3.0 57 140.84 7.17
8 3.5 65 160.61 8.18
9 4.0 71 175.44 8.94
10 4.5 77 190.26 9.69
11 5.0 83 205.09 10.45 105 9.95
12 5.5 86 212.50 10.82
13 6.0 89 219.92 11.20
14 6.5 91 224.86 11.45
15 7.0 92 227.33 11.58
16 7.5 93 229.80 11.71 134 8.73
17 8.0 94 232.27 11.83
18 8.5 94 232.27 11.83

30
Table 3.13 CBR test for expansive soil + 20% MICP by soil weight
[

Standard
Plunger Dial CBR CBR
load
penetration gauge Applied
stress intensity intensity
Sl. No. (𝒎𝒎) readings load (𝒌𝒈)
(𝒌𝒈/𝒄𝒎𝟐) (𝒌𝒈/𝒄𝒎𝟐) (% 𝒂𝒈𝒆)

1 0.0 2 4.94 0.25

2 0.5 7 17.30 0.88
3 1.0 17 42.01 2.13
4 1.5 28 69.19 3.51
5 2.0 38 93.90 4.77
6 2.5 46 113.67 5.77 70 8.25
7 3.0 52 128.49 6.53
8 3.5 58 143.32 7.28
9 4.0 63 155.67 7.91
10 4.5 68 168.03 8.53
11 5.0 72 177.91 9.04 105 8.61
12 5.5 76 187.80 9.54
13 6.0 80 197.68 10.04
14 6.5 83 205.09 10.42
15 7.0 86 212.51 10.79
16 7.5 89 219.92 11.17 134 8.34
17 8.0 91 224.86 11.42
18 8.5 93 229.80 11.67
19 9.0 94 232.27 11.80
20 9.5 94 232.27 11.80
21 10 94 232.27 11.80 162 7.28

31
Table 3.14 CBR test for expansive soil + 25% MICP by soil weight

Standard
Plunger Dial CBR CBR
load
penetration gauge Applied
stress intensity intensity
Sl. No. (𝒎𝒎) readings load (𝒌𝒈)
(𝒌𝒈/𝒄𝒎𝟐) (𝒌𝒈/𝒄𝒎𝟐) (% 𝒂𝒈𝒆)

1 0.0 2 4.94 0.25

2 0.5 5 12.36 0.63
3 1.0 14 34.59 1.76
4 1.5 23 56.83 2.90
5 2.0 33 81.54 4.16
6 2.5 41 101.31 5.16 70 7.37
7 3.0 48 118.61 6.04
8 3.5 55 135.91 6.93
9 4.0 61 150.73 7.68
10 4.5 66 163.09 8.31
11 5.0 71 175.44 8.94 105 8.51
12 5.5 75 185.33 9.44
13 6.0 79 195.21 9.95
14 6.5 82 202.62 10.32
15 7.0 86 212.51 10.83
16 7.5 89 219.92 11.21 134 8.36
17 8.0 90 222.39 11.33
18 8.5 91 224.86 11.46
19 9.0 91 224.86 11.46

32
 Variation of CBR values with MICP
18

16
CBR Intensity (% of age )

14

12

10

0
0 5 10 15 20 25 30
% of MICP

Graph 5 Plot variation of CBR values with MICP

3.4.5 Unconfined Compressive Strength (UCS)

3.14 UCS Testing Machine

33
 Table 3.15 UCS test for expansive soil only
[

Dial Proving Corrected Compressive

Sl. Deformation Load Strain
gauge ring area Strength
(𝒎𝒎) (𝒌𝑵) (%)
No. reading reading (𝒎𝒎𝟐) (𝑵/𝒎𝒎𝟐)
1 0 0.0 0 0 0 1133.54 0
2 50 0.5 14 0.019 0.6 1141.04 0.017
3 100 1.0 36 0.052 1.3 1148.65 0.044
4 150 1.5 69 0.098 1.9 1156.36 0.085
5 200 2.0 101 0.144 2.6 1164.17 0.123
6 250 2.5 111 0.158 3.3 1172.09 0.135
7 300 3.0 131 0.186 3.9 1180.12 0.158
8 350 3.5 149 0.212 4.6 1188.26 0.178
9 400 4.0 159 0.226 5.3 1196.51 0.189
10 450 4.5 166 0.236 5.9 1204.88 0.196
11 500 5.0 168 0.240 6.6 1213.36 0.197
12 550 5.5 169 0.241 7.2 1221.97 0.197
13 600 6.0 169 0.241 7.9 1230.70 0.196
14 650 6.5 168 0.240 8.5 1239.55 0.193
15 700 7.0 168 0.240 9.2 1248.53 0.191
16 750 7.5 167 0.238 9.8 1257.65 0.189
17 800 8.0 165 0.235 10.5 1266.89 0.185
18 850 8.5 165 0.235 11.2 1276.28 0.184
19 900 9.0 163 0.232 11.8 1285.80 0.180
20 950 9.5 162 0.231 12.5 1295.47 0.178

34
Table 3.16 UCS test for expansive soil + 5% MICP by soil weight

Dial Proving Corrected Compressive

Sl. Deformation Load Strain
gauge ring area Strength
(𝒎𝒎) (𝒌𝑵) (%)
No. reading reading (𝒎𝒎𝟐) (𝑵/𝒎𝒎𝟐)
1 0 0.0 0 0 0 1133.54 0
2 50 0.5 10 0.014 0.6 1141.04 0.012
3 100 1.0 24 0.034 1.3 1148.65 0.029
4 150 1.5 49 0.069 1.9 1156.36 0.060
5 200 2.0 67 0.095 2.6 1164.17 0.082
6 250 2.5 90 0.128 3.3 1172.09 0.109
7 300 3.0 101 0.144 3.9 1180.12 0.122
8 350 3.5 119 0.169 4.6 1188.26 0.142
9 400 4.0 126 0.179 5.3 1196.51 0.150
10 450 4.5 137 0.195 5.9 1204.88 0.162
11 500 5.0 144 0.205 6.6 1213.36 0.169
12 550 5.5 146 0.208 7.2 1221.97 0.170
13 600 6.0 169 0.241 7.9 1230.70 0.195
14 650 6.5 168 0.239 8.5 1239.55 0.193
15 700 7.0 168 0.239 9.2 1248.53 0.191
16 750 7.5 167 0.238 9.8 1257.65 0.189
17 800 8.0 165 0.235 10.5 1266.89 0.185
18 850 8.5 165 0.235 11.2 1276.28 0.184

35
Table 3.17 UCS test for expansive soil + 10 % MICP by soil weight

Dial Proving Corrected Compressive

Sl. Deformation Load Strain
gauge ring area Strength
(𝒎𝒎) (𝒌𝑵) (%)
No. reading reading (𝒎𝒎𝟐) (𝑵/𝒎𝒎𝟐)
1 0 0.0 0 0 0 1133.54 0
2 50 0.5 11 0.015 0.6 1141.04 0.013
3 100 1.0 35 0.049 1.3 1148.65 0.043
4 150 1.5 71 0.101 1.9 1156.36 0.087
5 200 2.0 98 0.139 2.6 1164.17 0.119
6 250 2.5 109 0.155 3.3 1172.09 0.132
7 300 3.0 132 0.188 3.9 1180.12 0.159
8 350 3.5 153 0.218 4.6 1188.26 0.183
9 400 4.0 164 0.233 5.3 1196.51 0.195
10 450 4.5 170 0.242 5.9 1204.88 0.201
11 500 5.0 173 0.246 6.6 1213.36 0.203
12 550 5.5 177 0.252 7.2 1221.97 0.206
13 600 6.0 177 0.252 7.9 1230.70 0.204
14 650 6.5 178 0.253 8.5 1239.55 0.204
15 700 7.0 176 0.250 9.2 1248.53 0.201
16 750 7.5 175 0.249 9.8 1257.65 0.198
17 800 8.0 175 0.249 10.5 1266.89 0.196
18 850 8.5 174 0.248 11.2 1276.28 0.194
19 900 9.0 173 0.246 11.8 1285.80 0.191

36
Table 3.18 UCS test for expansive soil + 15% MICP by soil weight

Dial Proving Strain Corrected Compressive

Sl. Deformation Load
gauge ring (%) area Strength
(𝒎𝒎) (𝒌𝑵)
No. reading reading (𝒎𝒎𝟐) (𝑵/𝒎𝒎𝟐)
1 0 0.0 0 0 0 1133.54 0
2 50 0.5 6 0.008 0.6 1141.05 0.007
3 100 1.0 24 0.034 1.3 1148.65 0.029
4 150 1.5 48 0.068 1.9 1156.36 0.059
5 200 2.0 71 0.101 2.6 1164.17 0.086
6 250 2.5 93 0.132 3.3 1172.09 0.113
7 300 3.0 107 0.152 3.9 1180.12 0.129
8 350 3.5 128 0.182 4.6 1188.26 0.153
9 400 4.0 141 0.201 5.3 1196.51 0.168
10 450 4.5 147 0.209 5.9 1204.88 0.174
11 500 5.0 150 0.213 6.6 1213.36 0.176
12 550 5.5 151 0.215 7.2 1221.97 0.176
13 600 6.0 151 0.215 7.9 1230.70 0.175
14 650 6.5 150 0.213 8.5 1239.55 0.172
15 700 7.0 149 0.212 9.2 1248.53 0.170
16 750 7.5 148 0.211 9.8 1257.65 0.167

37
Table 3.19 UCS test for expansive soil +20% MICP by soil weight

Dial Proving Strain Corrected Compressive

Sl. Deformation Load
gauge ring (%) area Strength
(𝒎𝒎) (𝒌𝑵)
No. reading reading (𝒎𝒎𝟐) (𝑵/𝒎𝒎𝟐)
1 0 0.0 0 0 0 1133.54 0
2 50 0.5 8 0.011 0.6 1141.05 0.01
3 100 1.0 21 0.030 1.3 1148.65 0.026
4 150 1.5 46 0.065 1.9 1156.36 0.057
5 200 2.0 71 0.101 2.6 1164.17 0.087
6 250 2.5 92 0.131 3.3 1172.09 0.111
7 300 3.0 114 0.162 3.9 1180.12 0.137
8 350 3.5 132 0.188 4.6 1188.26 0.158
9 400 4.0 140 0.199 5.3 1196.51 0.166
10 450 4.5 144 0.205 5.9 1204.88 0.170
11 500 5.0 145 0.206 6.6 1213.36 0.170
12 550 5.5 145 0.206 7.2 1221.97 0.169
13 600 6.0 145 0.206 7.9 1230.70 0.168
14 650 6.5 144 0.205 8.5 1239.55 0.165
15 700 7.0 143 0.203 9.2 1248.53 0.163

38
Table 3.20 UCS test for expansive soil + 25% MICP by soil weight

Dial Proving Corrected Compressive

Sl. Deformation Load Strain
gauge ring area Strength
(𝒎𝒎) (𝒌𝑵) (%)
No. reading reading (𝒎𝒎𝟐) (𝑵/𝒎𝒎𝟐)
1 0 0.0 0 0 0 1133.54 0
2 50 0.5 12 0.017 0.6 1141.05 0.015
3 100 1.0 36 0.051 1.3 1148.65 0.044
4 150 1.5 69 0.098 1.9 1156.36 0.085
5 200 2.0 92 0.1311 2.6 1164.17 0.112
6 250 2.5 104 0.148 3.3 1172.09 0.126
7 300 3.0 124 0.176 3.9 1180.12 0.149
8 350 3.5 130 0.185 4.6 1188.26 0.156
9 400 4.0 139 0.198 5.2 1196.51 0.165
10 450 4.5 141 0.201 5.9 1204.88 0.166
11 500 5.0 142 0.202 6.5 1213.36 0.166
12 550 5.5 142 0.202 7.2 1221.97 0.165
13 600 6.0 140 0.199 7.9 1230.70 0.162
14 650 6.5 140 0.199 8.5 1239.55 0.16
15 700 7.0 139 0.198 9.2 1248.53 0.158
16 750 7.5 138 0.196 9.8 1257.65 0.156

39
 Comparison of results from UCS Test
0.25
UCS Strength (N/mm^2)
0.2

0.15

0.1

0.05

0
0 2 4 6 8 10 12 14
Strain %

Only soil soil+5%of MICP Soil+10% of MICP

Soil+15% of MICP Soil +20% of MICP Soil+25% of MICP

Graph 6 Plot comparison results from UCS Test

variation of UCS with MICP

0.25

0.2

0.15
UCS Strength (N/mm^2)

0.1

0.05

0
0 5 10 15 20 25 30
MICP % in soils

Graph 7 Plot variation of UCS with MICP

40
 CHAPTER 4
RESULTS
Table 4.1 Comparison of Soil properties mixed with and without % of MICP

Soil+5% Soil+10% Soil+15% Soil+20% Soil+25%

S No Property Soil of of of of of
MICP MICP MICP MICP MICP
1
Atterberg limits
Liquid Limit 62.3 59.4 52.6 49.9 46.7 43.2

Plastic Limit 34.6 33.2 30.2 29.5 28.4 26.3

Plasticity Index 27.7 26.2 22.4 20.4 18.3 16.6

2
Standard proctor compaction test
a) MDD(g/cm3) 1.52 1.5 1.52 1.53 1.47 1.42

b) OMC(%) 22.62 22.13 22.56 21.27 23.57 21.34

3
California Bearing Ratio
Intensity (%) 5.99 10.58 16.67 9.95 8.61 8.51

4
Unconfined Compressive Strength
Strength(N/mm2) 0.197 0.195 0.206 0.176 0.17 0.166

41
 CHAPTER 5

CONCLUSION

Based on the results obtained & comparison made in the present study.

1. With the increasing bacterial percentage in the soil-MICP, the decrease in

value of plasticity was observed. Plasticity index values are directly
proportional to percent swell in an expansive soil, thus affecting the swelling
behavior of the soil-MICP.

2. The Maximum Dry Density (MDD) value of the black cotton soil initially
decreased with the addition of Bacteria. Then, it showed increment with
increasing Bacteria % in the soil- mixture. The maximum value of MDD
was observed for a mixture of soil and 15% of Bacterial percentage by
weight. The MDD values consistently decreased thereafter.

3. The Unconfined Compressive Strength (UCS) of the soil with variation of

bacterial percentage showed similar trend as that of the MDD values, except
the fact that the peak value was observed for a Bacteria percentage 10 % by
weight.

4. In California Bearing Ratio (CBR) tests of soil conducted with varying

MICP, the CBR increased gradually with the increase in MICP %. Till it
10% thereafter decreased.

5. Thus, MICP is an additive decreases the swelling, and increases the strength
of the black cotton soil.

Finally, we conclude that 10-15% of bacteria by weight of soils is sufficient for

soils to decrease the swelling (voids) and increase the strength of the soils.

42
 REFERENCES

➢ Achal, V., A. Mukherjee, P. C. Basu, and M. S. Reddy. 2009. Lactose

mother liquor as an alternative nutrient source for microbial concrete
production by Sporosarcina Pasteurian. Journal of Industrial
Microbiology and Biotechnology 36: 433–438.

➢ Dennis, M. L. and J. P. Turner. 1998. Hydraulic conductivity of

compacted soil treated with biofilm. Journal of Geotechnical and
Environmental Engineering 124(2): 120–127.

➢ DeJong, J. T., M. B. Fritzes’, and K. Nussle in. 2006. Microbially

induced cementation to control sand response to undrained shear.
Journal of Geotechnical and Geo-environmental Engineering 132(11):
1381–1392.

➢ 4. DeJong, J. T., B. M. Mortensen, B. C. Martinez, and D. C. Nelson.

2010. Bio-mediated soil improvement. Ecological Engineering 36:
197–210. Deutsche Sammlung von Micro-organisms und
Zellkulturen. Accessed 2011. http://www.dsmz.de/

➢ Hardcastle, J. H. (2003). Evaluation and treatment of expansive

volcanic soils US95, Owyhee County, Idaho. Moscow: University of
Idaho.

➢ Hames, F., & Verstraete, W. (2002). Key roles of pH and calcium

metabolism in microbial carbonate precipitation. Reviews in
environmental science and biotechnology, 1(1), 3-7.

➢ Whiff in, V. S., van Paassen, L. A., & Harks, M. P. (2007). Microbial
carbonate precipitation as a soil improvement technique.
Geomicrobiology Journal, 24(5), 417-423.

43
➢ Whiff in, V. S. (2004). Microbial CaCO3 precipitation for the
production of bio-cement (Doctoral dissertation, Murdoch University).

➢ Wiseman, G., Komornik, A. and Greenstein, J. (1985). Experience

with roads and buildings on expansive clays. Transportation Research
Record 1032, 60-67.

➢ Achal, V., Mukherjee, A., & Reddy, M. S. (2010). Microbial concrete:

way to enhance the durability of building structures. Journal of
materials in civil engineering, 23(6), 730-734.

➢ Bang, S. S., Galinat, J. K., & Ramakrishnan, V. (2001). Calcite

precipitation induced by polyurethane-immobilized Bacillus pasteurii.
Enzyme and microbial technology, 28(4), 404-409.

44
 Soil Stabilization by Bacterial Cementation

M Karthik Kumar Reddy1, V Srija2, K Shiva Kumar3, G Sannith Reddy4, V Krishna Sai

Assistant Professor 1, B. Tech Scholars

ABSTRACT In India nearly 51.8 million hectares of land area are covered with Expansive soil (mainly Black Cotton soil). The
property of these expansive soils, in general, is that they are very hard when in dry state, but they lose all of their strength when in
wet state. In light of this property of expansive soils, these soils pose problems worldwide that serve as challenge to overcome for
the Geotechnical engineers. One of the most important aspects for construction purposes is soil stabilization, which is used widely
in foundation and road pavement constructions this is because such a stabilization regime improves engineering properties of the
soil, such as volume stability, strength and durability. In this process, removal or replacing of the problematic soil is done
replacement is done by a better-quality material, or the soil is treated with an additive. In the present study, using
MICP(Microbiological Induced Calcite Precipitation). stabilization of black cotton soil obtained from Huzurabad, Telangana state
is attempted. With various proportions of this additive i.e 5%, 10%, 15%, 20% & 25%, expansive soils is stabilized. plasticity index
(P.I.) of clay-MICP mixes show a decrease in value with increasing MICP content. In conclusion, addition of MICP results in
decreases the plasticity of the expansive soil, and increase in workability by changing its grain size and colloidal reaction. the UCS
& CBR values of clay with MICP mixes were observed. Analysis of the formerly found result exposes the potential of MICP as an
additive that could be used for improving the engineering properties of expansive soils.

Keywords : Soil stabilization, Black cotton soil, CBR,UCS.

This technique employs microbes as a primary factor

1 INTRODUCTION
The need for stabilizing soils becomes necessary
mainly because of two reasons: I) weak or inconsistent
soil properties and ii) need for urbanization especially
in areas with problematic soils such as expansive or
high plasticity clays. These highly plastic soils cause
heaving on the ground surface (volume change) with
change in moisture content. The change in moisture
content could be due to seasonal or climatic variations
and evapotranspiration of vegetation. Structures built
on expansive soils tend to undergo moderate to severe
cracking problems .In particular, lightly loaded
structures such as one or two story residential and
industrial structures and pavements often experience
severe damage associated with substantive repair and
mitigation costs.
Physical properties of soil can be modified by the use
of mechanical compaction and/or compaction
grouting while chemical properties of soil can be
modified by the use of chemical stabilizers such as
Portland cement, lime and fly ash. Mechanical
compaction is recommended for sandy soils and is
effective or economical to a depth less than 10 m.
Chemical stabilization is typically recommended for
expansive soils. Environmentally safe techniques such
as pre-wetting and moisture barriers are only possible
for small confined spaces, and are not suitable for
larger construction projects such as highways and
railways which spread for miles. As mentioned above,
artificial cementation techniques are not always
feasible and environmentally friendly. However,
reduction in the use of artificial cementation
techniques can be practiced by substituting with
environmentally friendly techniques or materials. One
such method of soil stabilization technique is,
Microbial Induced Calcite Precipitation (MICP).
1
for stabilization. Successful implementation of MICP
will have its application in a wide variety of civil
engineering fields such as stability of retaining walls,
embankments and dams, controlling soil erosion,
stabilizing cohesionless soils to facilitate the stability
of underground constructions, increasing bearing
capacity of shallow and piled foundation and reducing
the liquefaction potential of soils (Kucharski, Cord-
Ruwisch, Whiff in & Al-thaw Adi, 2012; Ivanov &
Chu, 2008).
1.1 EXPANSIVE SOILS
Expansive soils, which are also called as swell-shrink
soil, have the tendency to shrink and swell with
variation in moisture content. As a result of this
variation in the soil, significant distress occurs in the
soil, which is subsequently followed by damage to the
overlying structures. During periods of greater
moisture, like monsoons, these soils imbibe the water,
and swell; subsequently, they become soft and their
water holding capacity diminishes. As opposed to this,
in drier seasons, like summers, these soils lose the
moisture held in them due to evaporation, resulting in
their becoming harder. Generally found in semi-arid
and arid regions of the globe, these types of soils are
regarded as potential natural hazard – if not treated,
these can cause extensive damage to the structures
built upon them, as well causing loss in human life.
Soils whose composition includes presence of
montmorillonite, in general, display these kinds of
properties. Tallied in billions of dollars annually
worldwide, these soils have caused extensive damage
to civil engineering structures. Also called as Black
Cotton soils or Regur soils, expansive soils in the
Indian subcontinent are mainly found over the Deccan
trap (Deccan lava tract), which includes Maharashtra,
Andhra Pradesh, Gujarat, Madhya Pradesh, and some
scattered places in Odisha. These soils are also found
in the river valley of Narmada, Tape, Godavari and
Krishna. The depth of black cotton soil is very large in
the upper parts of Godavari and Krishna, and the
north-western part of Deccan Plateaut. The colour of
this soil varies from black to chestnut brown. 20% of
the total land area, on an average, of this country is
roofed by expansive soils. In the semi-arid regions,
just in the last couple of decades, damages due to the
swelling- shrinking action of expansive soils have
been observed prominently in form of cracking and
break-up of roadways, channel and reservoir linings,
pavements, building foundations, water lines,
irrigation systems, sewer lines, and slab-on-grade
members.
1.2 MICP APPLICATIONS
Microbes are often responsible for the chemical
cementation of soils in nature due to the precipitation
of cementing materials into the voids of soils and
rocks (Ivanov & Chu, 2008). Microbes are able to
precipitate cementing materials such as calcium,
magnesium, iron, manganese and aluminium which
are crystallized to form carbonates, silicates,
phosphates, sulphides and hydroxides (DeJong,
Fritzes’ & Nüsslein, 2006). The prime role of
microbes in the precipitation of minerals is their ability
to create an alkaline environment through various
physiological activities (Douglas & Beveridge, 1998).
Calcium carbonate (calcite) precipitation is observed
to be a general mineral precipitation process in the
microbial world under ambient environment (Bang,
Galina & Ramakrishnan, 2001). Soil stabilization via
MICP is one of several applications of bio-remediated
processes. MICP can be used for the elimination of
soluble calcium from wastewater generated by
industries. The high calcium concentration in water
can clog pipes and malfunction reactors.
MICP technique is considered to be a better and more
environmentally friendly alternative to the
conventional technologies. However, more
investigations are needed to properly understand the
possibilities and limitations. Further, its application in
effectively stabilizing expansive soils is still a
hypothesis and this research is an initial step in
evaluating this hypothesis and understanding the
applicability of MICP technique to stabilize expansive
soils.
2 LITERATURE REVIEW
Krishna Kumari (2021) The challenges to enhance
the weak soil constantly lay a need for research and
exploration to flourish a new and advance method for
soil stabilization. The feasible technique is Bacterial
cementation which has evolved recently and raised a
sustainable technique for soil improvement. To
analyse the potency of MICP (Microbiologically
induced calcium carbonate precipitation) in
developing shear strength and lowering the hydraulic
conductivity of soil. Laboratory has been done to
2
certify the effect of Bio grout (Bacterial Cementation)
process on the strength of 2 types of sand (High Plastic
clay and Low plastic clay). The species of Bacillus
group, Magisterium was utilized to activate the calcite
precipitation. The strain of Bacillus group was utilized
to trigger and catalyse the calcite precipitation due to
chemical reaction of calcium chloride and urea. MICP
utilizes bacteria to hydrolyse urea and provide
carbonate ions and that reacts with the calcium
chloride solution which provides calcium carbonate
(calcite) that binds and holds the soil particles together
leads to increase the strength of soils and raise
stiffness. It was also found that the strength developed
with advancement in treatment duration. The outcome
of this experiment was evidently shows from the
pictures of Scanning Electron Microscope (SEM).
Yasin Dursun Sari 2014 Civil Engineering
Department, Faculty of Engineering and Architecture,
Istanbul Gallisin University, Istanbul, Turkey 6
August 2014 the biologically induced cementation
(Bio Grout) method has been used in this study to
improve engineering properties of soil. Laboratory
tests have been carried out to quantify the effect of Bio
Grout treatment on the strength of two types of sand
by using Sproutarian Pasteurian bacteria. Bearing
strength and other parameters, such as colony forming
units (number of bacteria), pH level, temperature, and
amount of CaCO3, have also been studied.
Calcification (cementation) through precipitation of
CaCO3 on the mineral surfaces has been clearly
observed from the images of a scanning electron
microscope (SEM). Finally, the effect of sand material
on the effectiveness of bio cementation was
investigated via employment of different sand types.
Masharu Fukue, Shin-Ich Ono and Yoshio Sato
2011 Microbial precipitation of carbonate was
observed using high microbial urease activity, and it
was found that the ratio of Mg/Ca asserted the types of
crystals produced. Without Mg2＋, calcite was
produced using only CaCl2, while the presence of Mg
produced Mg-calcite, magnesite and/or possibly
dolomite of round, spherical or bores shapes,
depending on reaction time, pH and Mg/Ca ratio. The
carbonate produced contributed to the development of
cementation for sands. The presence of Mg showed a
relatively strong cementation of the carbonate.
3 Materials & Methodology
3.1 Bacteria
Bacteria are microscopic organisms, single-celled
prokaryotic creatures. Bacteria come in different
shapes and the sizes. Bacteria are ubiquitous in every
habitat on Earth, growing in soil, acidic hot springs,
radioactive waste, water, and deep in the Earth’s crust,
as well as in organic matter and the live bodies of
plants and animals. There are typically 40 million
bacterial cells in a gram of soil and a million bacterial
cells in a milliliter of fresh water; in all, there are
approximately five nonillion (5×1030) bacteria on
Earth (Whitman et al. 1998, Vol.95) forming much of
the world’s biomass. Bacteria were first observed by
Antoine van Leeuwenhoek in 1676, using a single-
lens microscope of his own design. He called them
“animalcules” and published his observations in a
series of letters to the Royal Society. The name
bacterium was introduced much later, by Christian
Gottfried Ehrenberg in 1838.
There are broadly speaking two different types of cell
wall in bacteria, called Gram- positive and Gram-
negative. The names originate from the reaction of
cells to the Gram stain, a test long employed for the
classification of bacterial species.
3.2 Bacillus subtillus
Researchers with different bacteria proposed different
bacterial cementation in soils. The various bacteria
used in the bacterial cementation are Bacillus
pasteurii, Bacillus sphearicus, E. coli etc. In the
present study an attempt was made by using the
bacteria Bacillus subtilis strain no. JC3. The main
advantage of embedding bacteria in the soils is that it
can constantly precipitate calcite. This phenomenon is
called microbiologically induced calcite precipitation
(MICP). Calcium carbonate precipitation, a
widespread phenomenon among bacteria, has been
investigated due to its wide range of scientific and
technological implications. Bacillus subtilis is a
laboratory cultured soil bacterium and its effect on the
strength and durability is studied here.
3.3 Methodology
The present work is divided into two phases,
Phase I – Developing Culture of bacteriaPhase II–
Conducting tests in Laboratory, i.e., Grain size
distribution, Liquid limit, Plastic limit, CBR, UCS,
etc.
Phase I – Developing Culture of bacteria
Petri Dishes, Glass Tubes, Conical Flasks are sanitized
using the Autoclave machine in the laboratory. Raw
Bacteria Bacillus Subtillus is brought from the market.
Bacteria culture is developed by taking a small amount
of the raw bacteria under the UV radiation zone in the
pharmacy laboratory in our institution. Distilled Water
and Food for bacteria is taken with different chemicals
mixed with the distilled water.
The chemicals which are used as food nutrient for the
development of bacteria culture is that:
In a 250ml of conical flask, the following are taken:
Beef extract powder- 1%
Peptone- 1%
Sodium chloride- 0.5%
3
Agar agar Powder – 2%
This mixture is stored in a 10ml glass tubes and raw
bacteria of 1 ml is inserted into the glass tubes. These
tubes are done again sterilization in the Autoclave
machine at a pressure of 50lb/in2 by keeping the
cotton plugs such that no liquid comes out it when high
pressure is applied in the machine. Then, this liquid is
applied on the petri dishes and waited until it becomes
a solid (in the form of a wax). Streaks are drawn on it
so that to identify the development of culture of the
bacteria. These petri dishes are placed in the BOD
incubator for 48 hours and the bacteria culture is
developed after the removal of the petri dishes from
the Incubator. The developed bacteria can be seen with
the naked eye which forms in a white color. he
developed culture is taken with a small amount using
stirrer and placed in the required quantity of water that
is used in the bacterial cementation production. A
conical flask is taken and distilled water poured into it
and a small number of bacteria is inoculated into the
flask. This flask is again placed in the Autoclave
machine for the proper mixture of bacteria into the
water.
Adding shows
Requir Culturi
the UV Growth
ed
chemic
steriliz
chamb
ng the
of
Chemi ation bacteri
cals
als in er
a
bacteri
flask a
Fig 1 Bacteria culture
Phase II – Conducting tests in Laboratory
i.e., Grain size distribution, Liquid limit, Plastic limit,
CBR, UCS, etc.…
B5: 5% Bacteria by weight of soil
B10: 10% Bacteria by weight of soil
B15: 15% Bacteria by weight of soil
B20: 20% Bacteria by weight of soil
B25: 25% Bacteria by weight of soil
The above tests are done with proper attention and
record the values. Compare the bacterial Cementation
recorded values with without Bacterial cementation
values and plot the graph between them.
3.4 Methodology Adopted Graph 1 Sieve Analysis Graph

To evaluate the effect of MICP as a stabilizing 70

additive in expansive soils, series of tests, where the Sieve Analysis
content of MICP in the expansive soil was varied in 60
values of 5% to 25% (multiples of 10) by weight of
the total quantity taken. The Indian Standard codes 50
were followed during the conduction of the following
experiments:

% of finer
40

Standard proctor test – IS 2720 (Part 7) – 1980

30
Unconfined compressive strength (UCS) test – IS
2720 (Part 10) – 1991 20

California bearing ratio (CBR) test – IS 2720 (Part 16) 10

– 1987
0
Liquid & Plastic limit test – IS 2720 (Part 5) – 1985
0 0.2 0.4 0.6 0.8 1 1.2
Sieve size
4.2 plasticity index with Bacteria in expansive soil.
4 RESULTS Table 2 Variation of plasticity index with Bacteria
in expansive soil.
4.1 Sieve Analysis
Mixture Plasticity
Liquid limit Plastic limit
Table 1 Sieve Analysis index

Only soil 62.3 34.6

Sieve Weight Cumulative Cumulative % 27.7
size Retained weight % retained Finer
Soil + 5% 59.4 33.2
retained 26.2

Soil + 10% 52.6 30.2 22.4

4.75 83 83 27.67 72.33
Soil + 15% 49.9 29.5 20.4
1 19 102 34 66
Soil + 20% 46.7 28.4
0.6 75 177 59 41 18.3

0.425 13 190 63.33 36.67 Soil + 25% 43.2 26.3 16.9

0.3 34 224 74.67 25.33

0.212 17 241 80.33 19.67

Graph 2 Plot of variation plasticity Index with MICP
0.15 18 259 86.33 13.67

0.075 31 290 96.67 3.33 variation of plasticity index with MICP

30
pan 10 300 100 0
25
Plasticity Index

20

15

10

0
0 10 20 30
% of MICP

4
 Graph 6 Plot variation of UCS with MICP
Graph 3 Plot Variation of MDD with MICP
variation of UCS with MICP
0.25

UCS Strength (N/mm^2)

Variation of MDD with MICP 0.2
1.55 0.15
0.1
1.5
0.05
Yd

1.45 0
0 10 20 30
MICP % in soils
1.4
0 10 20 30
% of MICP
Table 3 Comparison of Soil properties mixed with and
without % of MICP
Yd
Soil Soil Soil Soil Soil
S Propert So +5 +10 +15 +20 +25
Graph 4 Plot Variation of OMC with MICP
N y il % % of % % of % of
o of MIC of MIC MIC
MI P MIC P P
Variation of OMC with MICP CP P
25 1 Standard proctor compaction test
Moisture content %

20 a MDD(g/ 1. 1.5 1.52 1.53 1.47 1.42

cm3) 52
15 b OMC(% 22 22.1 22.5 21.2 23.5 21.3
) .6 7
10
2 California Bearing Ratio
5
Intensity 5. 10.5 16.6 9.95 8.61 8.51
0 (%) 99 8 7
0 10 20 30
3 Unconfined Compressive Strength
% of MICP by soil weight
Strength 0. 0.19 0.20 0.17 0.17 0.16
(N/mm2 19 5 6 6 6
) 7
Graph 5 Plot Variation of CBR values with MICP

5 CONCLUSION
Variation of CBR values with MICP Based on the results obtained & comparison made in
the present study.
20
CBR Intensity (% of age )

1.The MDD was observed for mixture of soil and 15%

15
of bacteria. The MDD values consistently decreased
10 thereafter.

5 2.The bacteria is an additive material it decreases

swelling and increase the strength of the soil.
0
0 10 20 30 3.In California Bearing Ratio (CBR) tests of soil
% of MICP conducted with varying MICP, the CBR increased
gradually with the increase in MICP %.Till it 10%
thereafter decreased.
4.The variation of UCS of the soil with MICP were
observed in the 10% of MICP to the weight of soil.

5
REFERENCES
1. Achal, V., A. Mukherjee, P. C. Basu, and M. S.
Reddy. 2009. Lactose mother liquor as an
alternative nutrient source for microbial concrete
production by Sporosarcina Pasteurian. Journal of
Industrial Microbiology and Biotechnology 36:
433–438.
2. Dennis, M. L. and J. P. Turner. 1998. Hydraulic
conductivity of compacted soil treated with
biofilm. Journal of Geotechnical and
Environmental Engineering 124(2): 120–127.
3. DeJong, J. T., M. B. Fritzes’, and K. Nussle in.
2006. Microbially induced cementation to control
sand response to undrained shear. Journal of
Geotechnical and Geo-environmental
Engineering 132(11): 1381–1392.

4. Whiff in, V. S., van Paassen, L. A., & Harks, M.

P. (2007). Microbial carbonate precipitation as a
soil improvement technique. Geomicrobiology
Journal, 24(5), 417-423.
5. Whiff in, V. S. (2004). Microbial CaCO3
precipitation for the production of bio-cement
(Doctoral dissertation, Murdoch University).
6. Wiseman, G., Komornik, A. and Greenstein, J.
(1985). Experience with roads and buildings on
expansive clays. Transportation Research Record
1032, 60-67.
7. Achal, V., Mukherjee, A., & Reddy, M. S. (2010).
Microbial concrete: way to enhance the durability
of building structures. Journal of materials in civil
engineering, 23(6), 730-734.
8. Alexander, M. (1961). Introduction to soil
microbiology. Wiley, New York.