2.1.

INTRODUCTION:

The process of improving the strength and durability of soil is known as soil
stabilization. The main aim of stabilization is cost reduction and to efficiently use the locally
available material. The objectives of any stabilization technique used are to increase the
strength and stiffness of soil, improve workability and constructability of the soil and reduce
the Plasticity Index. For any given soil many stabilization methods, using different stabilizing
agents, may be effective to improve the soil properties in-place rather than removing and
replacing the material. Most common application of stabilization of soil is seen in
construction of roads and airfields pavement.
There are various methods of soil stabilization based on additives using to stabilize
the ground. Following are the commonly used stabilization techniques
 Mechanical stabilization
Mechanical stabilization is done by using rollers, compactors, vibrators.
 Chemical stabilization
Chemical stabilization is done by adding chemical additives to the soil that physically
combines with soil particles and alter the geotechnical properties of soil.
 Bio enzymatic stabilization
Enzymatic stabilization is a non-traditional stabilization method. This stabilization is
done by adding enzymes to the soils.
 Microbial stabilization
Microbial stabilization is also a non-traditional stabilization method. This stabilization
is done by adding bacteria and giving nutrient to the bacteria.

2.2. Concept of bio enzyme

Bio enzymes are catalytic enzymes that speed up chemical reactions they work extremely fast
in anaerobic or aerobic aqueous environment. Many materials change their form over a
period of time but transformations that would take years are accomplished within a fraction
of a second with the appropriate catalyst. Enzymes are protein molecules that speed up
chemical reactions some enzymes break down complex compound to form simpler ones,
while others build complex compounds from simpler ones. A single enzyme can perform its
entire function a million times a minute therefore chemical reactions occur thousands and
millions of time faster with an enzyme.
 It is a natural, non-toxic, non-flammable, non-corrosive liquid enzyme formulation
fermented from vegetable extracts that improves the engineering qualities of soil, facilitates
higher soil compaction densities, and increases stability. Enzymes catalyse the reactions
between the clay and the organic cat-ions and accelerate the cat-ionic exchange process to
reduce adsorbed layer thickness. For other types of chemical stabilization, chemicals are
mixed with soil, which is difficult to mix thoroughly, but Bio-enzyme is easy to use as it can
be mixed with water at optimum moisture content and then it is sprayed over soil and
compacted.
2.3. Bio-enzyme as soil stabilizer in road construction
Bio enzyme during soil stabilization breaks down the electric double layer between
the clay and static water. By this the clay particles looses its inherent charge and looses the
adhered static water layer. In this mechanism the clay particles segregate and are so fixed
crystallographically that it prevents any further volume changes on exposure to water.
The other aspect of bio enzymes of the newly developed variety is to block the micro
pores in the clay particles which gives it a large surface area and crevices for anions to attach
itself this prevents the clay particle from forming any further double layers. These larger
aggregates that are segregated together to form macromolecules are bound together
chemically. This chemical bond is relatively more flexible than that formed by cement and
lime stabilization hence can take increased repetitive loading constantly. Since all or most of
the plant extracted enzymes are produced by fermentation process, that utilizes
microorganisms to produce enzymes, they always have many materials which reduce voids in
the soil they also help to maintain the moisture balance in the soil thus it results in a
substantial cost savings on watering of the stabilized soil. This water retaining capacity not
only bonds the soil but also greatly reduces the suspended particulate matter in the
surrounding area of an unpaved road.
Bio enzymes are chemical, organic, and liquid concentrated substances which are
used to improve the stability of soil sub-grade for pavement structures. Bio-Enzyme is
convenient to use, safe, effective and dramatically improves road quality.
LACUOTURE AND GONZALEZ (1995) conducted a comprehensive study of the
TerraZyme soil stabilizer product and its effectiveness on sub-base and sub-grade soils. The
variation in properties was observed over a short period only and it was found that in
cohesive soils there was no major variation in properties during the early days but the soil
showed improved performance progressively.
HITAM AND YUSOF (1998) of Palm Oil Research Institute of Malaysia conducted field
studies on improvement on plantation roads. TerraZyme was treated to 27.2 km of the road,
which was having serious problems during the monsoon season or after heavy downpour. The
sections were then monitored on the surface erosion due to rainwater and wear due to usage.
After two monsoon seasons the road was found to be in very good condition in spite of large
exposure to heavy rainfall. No surface damage was observed, thus requiring no repair works
to the road section. TerraZyme stabilization can convert the road to an all-weather road that
has minimum destruction in hot and wet season.

BRAZETTI AND MURPHY (2000) conducted field experiments in Brazil to study the use
of TerraZyme as the bio-enzyme stabilizer for road construction. The selected soils were
sandy clay, silty clay, sandy silt, plastic and non-plastic clay, sandy loam, loam mixed with
clay, soil mixtures with pieces of recycled pavement. The field stretches were periodically
tested with DCP (Dynamic Cone Penetrometer) equipment. After the evaluation it was
concluded that the enzyme stabilization is a good technique for the effective and economic
solution for pavement construction.

SHARMA (2001) has conducted laboratory studies on use of Bio-Enzyme stabilization of

three types of soils namely, clay of high plasticity, clay of low plasticity and silt of low
plasticity. It was found that soil shows a marginal improvement in CBR value and
substantiates reduction in saturation moisture after four weeks of stabilization. The soil shows
a marginal improvement in Unconfined compressive strength.
ANDREW et al (2003) conducted laboratory scale testing program to evaluate the
effectiveness of enzyme treatment on sub grade soil. The effectiveness of enzyme treatment
was evaluated on the basis of statistical measurement of change in CBR, strength, soil
stiffness and soil modulus. The CBR test appears to be a relatively poor indicator of direct
soil strength for testing conditions.

ISAAC et al (2003) conducted a comprehensive study of the TerraZyme and its

effectiveness on lateritic soil and clay type soil collected from Kerala. The reactions of the
soils treated with the enzyme was observed and recorded and compared to the untreated
samples for the period of 8 weeks. It was found that in all soil types considered, the CBR
value has increased by addition of TerraZyme, which proved its suitability as a stabilizing
agent. The increase in CBR was of the range of 136 to 1800 times that of the original value.
TerraZyme is useful for clay soil and sand but is less significant to silty soils and clayey and
sandy soil had increase in CBR by 700 percent.

MANOJ et al (2003) conducted a study to assess the suitability of Bio-Enzyme as soil

stabilizer on five types of soils with low clay content to very high clay content. Laboratory
tests were conducted to determine the engineering properties of soil and strength
characteristics of soil with and without stabilization with Bio-Enzyme. The Bio-Enzyme
stabilization has shown little to very high improvement in physical properties of soil. This
little improvement may be due to chemical constituent of the soil, which has low reactivity
with Bio-Enzyme. In the cases of highly clay moderate soil, like silty soil to sandy soil, the
effect of stabilization has improved the CBR and unconfined compression strength.

MIHAI et al (2005) have done experiments on two types of soils with two enzyme products.
The soils are treated with two types of Enzyme with variable dosage 0.5 cc, 1cc, 1.5cc/5 l.
The result of the test indicated that soils treated with enzymes shows improvement in strength
and sustainability of soils. After the addition of enzyme the shear strength of soil also
increases considerably.
RAVI SHANKAR (2009) conducted study on Bio Enzyme stabilization as a highway
material on Lateritic soil and blended Lateritic soil are tested for engineering properties,
CBR, UCC with different dosages of Enzyme (Terrazyme). After treating of soils with
enzymes the tests was conducted it shows a medium improvement in physical properties with
enzyme dosage of 200ml/2m3. CBR value increased 300 % after curing of 4 weeks UCC
increased by 450% and permeability decreases by 42%.

SUREKA NAAGESH et al (2010) conducted a number of experiments on an expansive soil

treated with a bio enzyme. Soil Specimens were treated with different dosages of bio enzyme
and they concluded that soil specimens treated with bio enzyme exhibit lesser percent swell
and swell pressure compared to untreated soil specimens. Soils exhibited 29 % lesser swell
potential after 30 days curing.
VENKATASUBRAMANIAN & DHINAKARAN (2011) performed test on 3 different
soils with different properties. These soils were tested with different dosage of enzyme. The
liquid limit and plasticity index of soil were reported as 28, 30, 46 % and 6, 5 and 6 %
respectively. An increase of 157 to 673 % is seen in CBR after 4 weeks of curing and 152 to
200 % in UCC.
VENKATASUBRAMANIAN (2011) has conducted laboratory studies on effect of Bio
Enzymatic soil stabilization. Three different kinds of soils were taken and tested. In UCC
maximum rate of increase was observed with 21 days curing period
LEKHA (2013) has done experimental work on black cotton soil stabilization with
terrazyme. The black cotton soils are tested for engineering properties, CBR, UCC,
compaction, durability and Permeability. Changes are marginal for MDD of enzyme treated
soil whereas decrease in OMC is observed to be 13.46% from 17.08% after 8 weeks of
curing. The UCS value increases from 268 kN/m2 to 859 kN/m2 when compared to the
original soil after 8 weeks of curing period. The treated soaked CBR values are increased as
the curing periods increase. The percentage loss in weight of soil samples is reduced by 22%
for treated soil in comparison with untreated soil samples for freeze and thaw cycles, whereas
the soil samples could not withstand for wet and dry cycles.
SUNEET KAUR (2014) carried out a theoretical evaluation of enzyme. Reduction of about
18 to 26 % is seen in cost of construction of roads by using TerraZyme as a soil stabilizer,
constructed by public work department in Maharashtra. Structures made of bio enzyme are
economical and have greater strength.
PUNEET AGARWAL (2014) conducted study black cotton soil treated with Enzyme. UCC
test were conducted the relative rate of increase in the UCC value from 1 to 7 days of curing
is maximum at Terrazyme dosage of 1ml. Thus keeping economic criteria (getting maximum
strength at minimum cost) in mind the optimum dosage of Terrazyme for improving the UCC
of Black Cotton soil is about 1ml/5kg of soil.

VIJAY RAJORIA (2014) has been studied on stabilization of soil using permazyme.
PermaZyme is a compaction enzyme, when it is added to a soil and aggregate mixture; it
causes the compaction of clays and silts with a much faster rate than that occurs in nature. It
increases the lubricity of soil particles which allows the designated soil density to be reached
with less compactive effort. It promotes rapid saturation and inhibits surface evaporation,
thus reduces the requirement of water upto 25% of the O.M.C.
MADHAVI VEDULA (2014) has been studied on innovative rural road construction
techniques. In this Fujibeton the product is an inorganic polymer that chemically binds with
all compounds, where blended with ordinary Portland cement in 1 to 3% by weight of OPC.
The blended mix is called ‘Fujibeton Mix’, which is used for stabilization of soil that
improves the engineering properties of soil. Fujibeton improves CBR of the sub-grade and
does not create shrinkage cracks and is therefore highly effective for clayey/soils. With
Fujibeton, a high dry density is obtained with only minor compaction.
CHANDRASHEKAR (2014) has been studied on innovative rural road construction, in
these Renolith is used. Renolith is polymer based chemical, which is environmentally
friendly and which facilitates the bonding of soil particles. Soil-cement with Renolith has a
high modulus of elasticity and can disperse the wheel loads very effectively. The use of
Renolith, when used in soil stabilization with cement, gives strong and durable base
MANJUNATH (2014) studied on effect of dosage of bio enzyme on strength and plasticity
characteristics of soil. Two types of soils were considered clayey sand and highly
compressible clay (CH) respectively. Dosage of 2 ml/kg of dry soil increases UCC of clayey
sand and CH by 30% and 74% respectively. The effect of Terrazyme on UCC of black cotton
soil is more than clayey sand because of increased fines content
SWATHY (2015) has done experimental work on CH soil stabilization using enzyme
The soil properties have been much improved by stabilising with enzyme dosage of 0.2ml/kg.
The treated soil was observed to be having lesser plasticity index values. For the optimum
dosage of 0.ml/kg, the MDD of the soil increases up to 6% and OMC decreases up to 19%.At
the optimum dosage, there was an increase of 351% in the UCC strength of soil, Increase of
352% in the CBR value of soil.
VENIKA SAINI (2015) has done experimental studies on clayey silt (CM) treated with
enzyme. Different tests were conducted with and without treating of soil with enzymes.
2m3/200ml dosage of enzyme is the optimum one because the consistency limits are reduced
and the soaked CBR increased after curing period of two weeks.
SANDEEP PANCHAL (2017) conducted study on low compressible clayey soil treated
with Terrazyme with different curing periods. Different tests were conducted, CBR shows
improvement at 900ml/1m3 dosage of enzyme, curing period of 2 weeks. Engineering
properties and consistency limits shows an improvement in its properties after treating with
enzyme.

VENKATESH (2017) has conducted studies on black cotton soil used as subgrade material
with terrazyme. Shear Strength of the soil increased form 5.39 kPa at 0% Terrazyme to 27.5
kPa at 4% Terrazyme, percentage increase is 410%. With increase in percentage of
Terrazyme the un-soaked CBR value was increased from 3.93 to 8.03. Percentage increase is
104.32. Tri-Axial results showed with increase in percentage of Terrazyme, cohesion of the
soil increased from 6.85 kPa at 0% Terrazyme to 38.60 kPa at 4% Terrazyme, percentage
increase is 463.50.
1.1.MICROBIAL STABILIZATION
Microbial stabilization is an emerging method in Geotechnical Engineering that deals
with the application of Microbiological methods to improve the mechanical properties of soil
to make it more fitting or appropriate for construction and environmental purposes. In these
regard two noteworthy applications, Bio-clogging and Bio-cementation have been explored.
Bio-clogging is the production of pore-filling materials through microbial means so that the
porosity and hydraulic conductivity of soil can be reduced whereas Bio-cementation is the
generation of particle binding materials through microbial processes in situ so that the shear
strength of soil can be increased.

EDWARD.K AND ISMAIL (2008) conducted comprehensive study of microbiological

improvement of physical properties of soil. The applicability of microbiological process to
soil improvement will likely depend on a variety of factors including the type of microbial
metabolism.

F.D.MEYER S.BANG (2011) studied on effect of stabilisation of microbial induced soil.

Application of microbes for fugitive dust control. Environmentally friendly means to supress
airborne dust particles. The soil type and environment conditions play a significant role.

B.C.MARTINEZ AND T.R.GINN (2011) conducted experiment on upscaling of microbial

induced calcite precipitation in 0.5m columns modelling results were given. Numerical model
shows promising results for modelling microbial induced calcite precipitation. The regression
shows non uniqueness due to the lack of quantity and variety of data collected.

LIANG CHENG (2012) study on cementation of sand soil by microbial induced calcite
precipitation at various degree of saturation. Engineering properties of sand soil is varies
significantly, mainly because of the different location of the calcite deposited. The calcite
crystals formed under lower degree of saturation showed that more crystal are formed in the
contact points, where they contribute the most to the strength of the cemented samples.

AHMED AL QABANY (2012) study the factors affecting efficiency of microbial induced
calcite precipitation highlights the significance of treatment method to ensure that the
treatment used not only suits field conditions but also optional for the application it was
designed for and achieves the best possible results from the process.
BRENDAN C (2014) study on improving sand with microbial induced calcite precipitation.
SEM observations of treated sands were conducted. Higher cementation solution
concentrations were also found to produce greeted improvements in stiffness/ strength
particularly for non equimolar solutions

RAMAKRISHNAN R (2016) study on effect of microbial induced calcite precipitation on

strength of fine grained soils. Microbial induced calcite precipitation was found to increase
unconfined compressive strength of CI and CH soils. Microbial induced calcite precipitation
found to increase the undrained cohesion of soil. Strength parameter can be co related with
other properties and possible implication will further lead to increase bearing capacity,
minimised settlements, reduced permeability of soil.