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EXPERIMENTAL STUDY ON SELF HEALING

CONCRETE
1SURAJ BHAN SINGH SIKARWAR, 2Mr. Rameezut Tauheed

ABSTRACT
Cracking of concrete is a common phenomenon without immediate and proper treatment, cracks
in concrete structures tends to expand further and eventually require costly repairs. Even though
it is possible to reduce the extent of cracking by available modern technology, remediation of
cracks in concrete has been the subject of research for many years. Cracks and fissures are a
common problem in building structures, pavements, and historic monuments.

We have introduced a novel technique in fitting cracks with environmentally friendly biological
process that is a continuous self-remediating process. In this study bacillus substile that is
abundant in soil has been used to induced CaCo3. It is therefore vital to understand the
fundamentals of microbial participation in crack remediation. Cracks in concrete form an open
pathway to reinforcement can lead to durability problems like corrosion of the steel rebar’s.
furthermore, cracks can cause leakage in case of liquid retaining structures, due to alkali,
sulphate, and drying shrinkage to overcome this problem, a variant of smart concrete is rapidly
developing, which is known as “self-healing concrete”. The self-healing concrete is one that
senses its crack formation and react to cure itself without human intervention. The impact of
Eco-friendly micro-organism has been made use of the self-healing process in the current project.

The cement industry is a major global contributor to world CO2 emissions (7% in 2019). A major
cause of this high percentage is the durability issues associated with concrete, in recent years a
new breed on concrete that can heal cracks which are a major cause of these durability. This will
introduce this new breed of concrete in its various forms, with particular attention paid to the
form which incorporates use of microbes as the healing agents.

In recent years a bacteria-based self-healing concrete is being developed to extend the service
life. A two-component healing agent is added to the concrete mixture. The agent consists of
bacteria and mineral precursor compound. Whenever cracks occurs and water is present the
bacteria become active and convert the incorporated organic compounds into the mineral calcium
precipitates and can seal and block cracks, allowing autonomous healing. This paper aims to

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review the development of bacteria-based self-healing concrete, introducing the proposed

healing system. Different stages in the developing are discussed, and some recommendation for
further research.

Cement mortar durability is the function of its internal pore structure and distribution, porosity,
and its permeation properties. Research has shown that some specific bacterial species isolated
from soil can tolerate harsh and challenging alkaline environment and can be used in remediating
cracks in cement mortar structures. This state-of-the-art microbial based crack healing
mechanism is one such phenomenon on which studies were carried out to investigate the role of
calcite mineral precipitation in improvement of durability in bacteria integrated cement mortar.
The primary goal of this study is to explore the potential of biomineralization microbial calcium
carbonate deposition in cement mortar, to develop sustainable construction materials. The idea
has led to the conception of energy efficient and sustainable construction material called
‘Bacterial Cement mortar’. This paper primarily focuses on the studies related to the
characterization of bacteria produced calcium carbonate crystals using various nano
characterization techniques such as Scanning Electron Microscope (SEM), X-ray diffraction
(XRD), and Thermo gravimetric analysis (TGA) to validate that cracks/pore were sealed up by
calcite crystals grown due to complex metabolic mechanism of nitrogen cycle by Bacillus subtilis
JC3.

In concrete, cracking is a common phenomenon developed due to relatively low tensile strength.
High tensile strength may be developed in concrete due to external loads, imposed deformations,
plastic shrinkage, plastic settlement and expensive reaction. Proper and immediate treatment
should be done to prevent expansion of cracks which may eventually be of higher cost. For crack
repair, a variety of traditional repair system are available which possess several disadvantages
aspect such as different thermal expansion coefficient, environmental and hazards of health.
Bacterially induced calcium carbonate precipitation has been proposed as an alternative and
environmentally friendly crack repair technique. It is found that microbial mineral precipitation
as a result from metabolic activities of favorable bacteria in concrete improved the overall
behavior of concrete. It is expected that further development of this technique will result in a
more durable, sustainable, and crack free concrete that can be used efficiently for construction
in wet atmosphere where corrosion of reinforcement affects the durability, permeability, and
strength of concrete.

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CHAPTER - 1
INTRODUCTION

1.1 Definition: Self-healing concrete

The Self-healing Concrete is one that senses its crack formation and reacts to cure itself without
human intervention. Self-healing concrete is a product that will biologically produce limestone
to heal cracks that appear on the surface of concrete structures. Specially selected types of the
bacteria Bacillus subtilis, is added to the ingredients of the concrete when it is being mixed.
These self-healing agents can lie dormant within the concrete for up to 200 years.

1.2 Bacterial concrete

The “Bacterial Concrete” can be made by embedding bacteria in the concrete that are able to
constantly precipitate calcite. Bacillus subtilis is a soil bacterium, can continuously precipitate a
new highly impermeable calcite layer over the surface of an already existing concrete layer. The
favorable conditions do not directly exist in a concrete but have to be created. A Main part of the
research will focus on this topic. Tests are conducted to study the mechanical properties of the
above concrete with various percentages of Bacteria. The tests carried out are Compressive
strength test, Split Tensile strength test.

1.3 Mechanism

Some possible mechanisms for Self-healing are:

1. Formation of material like calcite.
2. Blocking of the path by sedimentation of particles.
3. Continued hydration of cement particles.
4. Swelling of the surrounding cement matrix

1.4 Objects of biological approach

To develop methods to enhance durability of concrete by adding bacteria. To regain the

maximum strength of concrete after cracking rectification. To develop methods of concrete after
cracking rectification.

1.5 Bio-mineralization

Natural processes such as weathering, faults, land subsidence, earthquakes and human activities
create fractures and fissures in concrete structures and historical stone monuments. These
fractures and fissures are detrimental since they can reduce the service life of the structure. In
the case of 20 monuments and buildings of historic importance, these cracks tend to disfigure
and destroy the structure. Use of bacterial concrete for remediating these structures will reduce
the crack width and increase the strength of the structure. Therefore, a novel technique for
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remediating damaged structural formations has been developed by employing a selective

microbial plugging process, in which microbial metabolic activities promote calcium carbonate
(calcite) precipitation. The technique is called “Microbiologically Enhanced Crack Remediation
(MECR). This technique comes under a broader category of science called “Biomineralization”.
It is a process by which living organism form inorganic solids. Bacterial deposition of a layer of
calcite on the surface of the specimens resulted in a decrease of capillary water uptake and
permeability towards gas. This bacterial treatment resulted in a limited change of the chromatic
aspect of mortar and concrete surface. The type of bacterial culture and medium composition had
a profound impact on CaCo3 crystal morphology.

Microbial mineral precipitation resulting from metabolic activities of some specific

microorganisms in concrete to improve the overall behavior of concrete has become a
considerable area of research. These bacteria can influence the precipitation of calcium carbonate
by the process of Ammonification (Ammo acid degradation). Precipitation of calcium carbonate
crystals occur by heterogeneous nucleation on bacterial cell walls once supersaturation is
achieved.

The application of concrete is rapidly increasing worldwide and therefore the development of
bacterial mediated concrete is urgently needed for environmental reasons. As presently, about
8% of atmospheric carbon dioxide emission is due to cement production, mechanisms that would
contribute to longer service life of concrete structures would make the material not only more
durable but also self-repair, i.e., the autonomous healing of cracks in concrete. The potential of
bacteria to act as self-healing agent in concrete has proven to be a promising future. This field
appears to be more beneficial as bacterial concrete appears to produce more substantially more
crack plugging minerals than control specimens (without bacteria). A promising sustainable
repair methodology is currently being investigated and developed in several laboratories, i.e., a
technique based on the application of mineral producing bacteria. The application for ecological
engineering purposes is becoming increasingly popular as is reflected by recent studies where
bacteria were applied for removal of chemicals from wastewater streams, for bioremediation of
contaminated soils and removal of greenhouse gases from landfills.

The applicability of specifically mineral producing bacteria for sand consolidation and limestone
monument repair and filling of pores and cracks in concrete have been recently investigated. In
all these studies so far, bacteria were externally applied on cracked concrete structures or test
specimens, i.e., as surface treatment or repair system.

An integrated healing agent would save manual inspection and repair and moreover increase
structure durability. Addition of such an agent to the concrete mixture would thus save both
money and the environment as less maintenance and use of environmentally friendly repair
material is needed. Microbial carbonate precipitation (bio deposition) decreases the permeation
properties of concrete. Hence, a deposition of a layer of calcium carbonate on the surface of
concrete resulted in a decrease of water absorption and porosity. Bacteria-based self-healing
concrete is produced by incorporating spores of bacteria of a special kind (Bacillus subtilisJC3),
in the concrete matrix at the stage of preparation of the concrete by mixing the spore suspension
in concrete mixing water. When crack is formed water enters the crack subsequently the
homogeneously distributed bacterial spore in hardened concrete matrix gets activated and
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germinate to become metabolically active vegetative cells that can convert the organic nutrient
compounds into insoluble inorganic calcium carbonate-based minerals.

Bio-mineralization is also defined as a biologically induced precipitation in which an organism

creates a local micro-environment with conditions that allow optimal extracellular chemical
precipitation of mineral phases. Numerous diverse microbial species participate in the
precipitation of mineral carbonates in various natural environments including soils, geological
formations, freshwater bio films, oceans, and saline lakes. The precise role of the microbes in
the carbonate precipitation process is still not clear. Almost all bacteria are capable of calcium
carbonate precipitation. A novel technique for the remediation of damaged structural formations
has been developed by employing a selective microbial plugging process, in which metabolic
activities promote precipitation of calcium carbonate in the form of calcite. Biomineralization of
calcium carbonate is one of the strategies to remediate cracks in building materials. In nature,
microorganisms can induce calcite mineral precipitation through nitrogen cycle by
ammonification of amino acids/ nitrate reduction/ hydrolysis of urea. The binding strength of the
precipitated crystals is highly dependent on the rate of carbonate formation and under suitable
conditions it is possible to control the reaction to generate hard binding calcite cement (or bio-
cement).

1.6 Chemistry of bio-classification by BACILLUS SUBTILIS JC3

Bacterial calcium carbonate precipitation results from both passive and active nucleation. Passive
carbonate nucleation occurs from metabolically driven changes in the bulk fluid environment
surrounding the bacterial cells. This increases the mineral saturation and induces nucleation. In
the Ammo acid degradation driven system, this occurs from an increase in pH due to
ammonification. Active carbonate nucleation occurs when the bacterial cell surface is utilized as
the nucleation site. The cell clusters exhibit a net electronegative charge which favors the
adsorption of Ca2+ ions. The Ca2+ ions attract CO3 2- and HCO3- ions, which will eventually
form calcium carbonate precipitates. Although it is known that there are many different types of
bacteria capable of calcium carbonate precipitation, it has been hypothesized that there are
specific attributes of certain bacteria that promote and affect CaCO3 precipitation more than
others. It has already been noted that cell walls have an inherent electronegative charge that affect
the binding of certain ions, but the extracellular polymeric substance associated with bio films
may also be involved. Bio film cells are contained in the extracellular polymeric substance matrix
and may exhibit an overall negative charge. This negative charge is important in trapping metal
ions. Strain Bacillus subtilis JC3, selected for the present study, was distinguished as aerobic
alkaliphilic spore-forming soil bacteria. The medium used to grow Bacillus subtilisJC3 was
based on peptone, NaCl, yeast extract. The pure culture was isolated from the soil sample of
JNTU.

Microbiologically induced calcium carbonate precipitation occurs via more complicated

processes than chemically induced precipitation. In nutrients medium, it is possible that
individual microorganisms produce ammonia because of amino acids degradation to create an
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alkaline micro-environment around the cell. The high pH of these localized areas, without an
initial increase in pH in the entire medium, commences the growth of CaCO3 crystals around the
cell. Specific proteins present in biological extracellular polymeric substances cause the
formation of different calcium carbonate polymorphs. Some bacteria and fungi can induce
precipitation of calcium carbonate extracellularly through several processes that include
photosynthesis, ammonification, denitrification, sulfate reduction and anaerobic sulphide
oxidation. Although all the Bacillus strains were capable of depositing calcium carbonate,
differences occurred in the amount of precipitated calcium carbonate on agar plate colonies.
Oxidative deamination of amino acids by Bacillus subtilisJC3 is temperature dependent and that
the highest calcite precipitation rates occurred near the point of critical saturation. B. SubtilisJC3
member of the genus Bacillus is Gram-positive, rod-shaped, endospore forming bacteria
commonly found in soil; precipitate calcium carbonate (CaCO3) in its micro-environment by the
ammonification of amino acids into ammonium (NH4+) and carbonate (CO3) ^2- ions.

Microbiologically induced (also called ‘‘bacteriogenic’’) calcite carbonate precipitation by

Ammonification (Ammo acid degradation) comprises of series of complex biochemical
reactions. Amino acids released during proteolysis (the process of enzymatic breakdown of
proteins by the microorganisms with the help of proteolysis enzymes) undergo deamination in
which nitrogen containing amino (NH2-) group is removed. Thus, process of deamination which
leads to the production of ammonia is termed as "ammonification".

The process of ammonification is mediated by Bacillus subtilisJC3. Ammonification usually

occurs under aerobic conditions (known as oxidative de amination) with the liberation of
ammonia (NH3) or ammonium ions (NH4) when dissolved in water. The biochemical reactions
of ammonification in peptone-based medium is represented as follows

Ca2+ + B.subtilisCell → B.subtilisCell- Ca2+CH3CH(NH2)COOH (Peptone) + ½O2 --------->

C2H2 + H2CO3 + NH3
H2CO3 ----------> H+ + HCO3-
NH3 + H2O --------> NH4+ + OH- (pH increase)
HCO3- + H+ + OH- ---------->CO3^2- + H2O
B.subtilis Cell- Ca2+ + CO3^2- → B.subtilis Cell- CaCO3

Upon examination, bacterial cells were shown encased in calcite crystals, which indicated that
the bacteria acted as a nucleation site for the mineralization process, an example of active
nucleation.

1.7 Application of bacteria in concrete

1.7.1 Microbial concrete as an alternative surface treatment for concrete

An important measure to protect concrete against damage is diminishing the uptake of water.
many of the physical and chemical deterioration mechanisms of concrete are related to aggressive
substances present in aqueous solution. Surface treatments play an important role in limiting the
infiltration of water. Broad arrays of organic and inorganic products are available in the market
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for the protection of concrete surfaces, such as a variety of coatings, water repellents and pore
blockers. But these means of protection beside their favorable influences even show
disadvantageous aspects such as: Degradation over time, need for constant maintenance,
Different thermal expansion coefficient of the treated layers, Use of certain solvents contributes
to environmental pollution as well.

1.7.2 Microbial concrete as concrete crack remediation/healing

When cracks appear in the concrete, the possibility for corrosion of the embedded steel arises
which could eventually ruin the integrity of the structure. Without immediate attention, the cracks
can expand and cause extensive damage. Current forms of concrete crack remediation are
structural epoxy, resins, epoxy mortar, and other synthetic filler agents. These synthetic solutions
often need to be applied more than once as the cracks expand.

Clearly there is a need for an effective, long-term, environmentally safe method to repair cracks
in concrete structures. Several research groups have investigated the possibility of
biomineralization as an effective method to remediate cracks and fissures in concrete structures.
Cracks filled with a mixture of B. subtilis, and sand showed a significant increase in compressive
strength and stiffness, compared to cracks without cells. Microscopy confirmed the presence of
calcite crystals and cells near the surface of the cracks.

CHAPTER - 2
LITERATURE REVIEW

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• V. Ramakrishnan, R.K.Panchalan, and S.S.Bang has published a paper on Bacterial Concrete

– A Concrete for the Future which says a common soil bacterium was used to induce calcite
precipitation. This technique is highly desirable because the mineral precipitation induced
because of microbial activities, is pollution free and natural. The effectiveness of this technique
was evaluated by comparing the compressive strength and stiffness of cracked specimens
remediated with bacteria and those of the control specimens (without bacteria). Experimental
investigation was also conducted to determine the strength regaining capacity (modulus of
rupture) of cracked beams remediated with different concentrations of bacteria. This paper also
presents the results of a durability study on cement mortar beams treated with bacteria, exposed
to alkaline, sulfate and freeze-thaw environments. Different concentrations of bacteria were used
for the investigation. It was found that the use of bacteria improved the stiffness, compressive
strength, modulus of rupture and durability of concrete. Scanning electron microscope (SEM)
was used to document the role of microbiologically induced mineral precipitation in improving
the strength and durability aspects of concrete.

• C. C. Gavimath, B. M. Mali, V. R. Hooli, J. D. Mallpur , A. B. Patil , D.P.Gaddi, C.R.Ternikar

has published a paper on potential application of bacteria to improve the strength of cement
concret in which the potential application of bacterial species i.e. B.sphaericus to improve the
strength of cement concrete is studied. Here they have made an attempt to incorporate dormant
but viable bacteria in the concrete matrix which will contribute to the strength of the concrete.
Water which enters the concrete will activate the dormant bacteria which in turn will give
strength to the concrete through the process of metabolically mediated calcium carbonate
precipitation. Concrete, however, is due to its high internal pH, relative dryness and lack of
nutrients needed for growth, a rather hostile environment for common bacteria, but there are
some extremophilic spore forming bacteria may be able to survive in this artificial environment
and increase the strength and durability of cement concrete. In this study they found that
incorporation of spore forming bacteria of the species Bacillus will not negatively affect the
compressive and split tensile strength of the cement concrete.

• Surendran and S. John Vennison has published a Journal on Occurrence and Distribution of
Mosquitocidal Bacillus sphaericus in Soil which says Bacillus sphaericus is one of the effective
biolarvicides to control Culex species and the monitoring of larval susceptibility is essential to
avoid resistance development. Mosquito larvicidal activity of B. sphaericus was assessed by
isolating them from ecologically different soil habitats in and around Devakottai of Tamil Nadu
inSouth India. The isolated organisms were
confirmed as Bacillus sphaericus based on biochemical characterization and microscopic
observations.

• Thirumalaichettiar has published a paper on bacterial concrete says a novel technique in

remediating cracks and fissures in concrete by utilizing microbiologically induced calcite
(CaCo3) precipitation is discussed. Microbiologically induced calcite precipitation (MICP) is a
technique that comes under a broader category of science called biomineralization. It is a process
by which living organisms form inorganic solids. Bacillus Pasteruii, a common soil bacterium
can induce the precipitation of calcite.

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• Ellie Zolfagharifard(2012) has published a article on Biological concrete could usher in a new
era of self-healing civil structures says its Far better would be to use a material that heals itself
just as a crack begins to appear. Existing research has focused on the use of synthetic materials
that can seal up cracks as they develop. But the work by Delft and Ghent universities is unique
in that they plan to use living bacteria to achieve what they hope will be better results. At Delft
University, DrHenk Jonkers has developed a biological concrete that uses specially selected
bacteria of the genus Bacillus, alongside a combination of calcium lactate, nitrogen and
phosphorus, to create a healing agent within the concrete. If untouched, these agents can remain
dormant in the concrete for centuries. But if water begins to seep into the cracks, the Emerging
Trends in Engineering Research 19 ISBN 978-93-82338-32-1| © 2012 Bonfring spores of the
bacteria start to generate and feed on the calcium lactate. This consumes oxygen, which in turn
converts the calcium lactate into limestone that solidifies and seals the surface. The removal of
oxygen also improves the durability of the steel reinforcement.

• Dr. Nele De Belie, Ghent University, Belgium has published a paper on Healing and Self-
Healing of Concrete has shown during their presentation how repair and consolidation of mineral
phases of building materials and the healing and self-healing of concrete with the help of bacteria
is possible. Micro-organisms play a crucial role in pedogenesis, transformation of minerals and
exchange of elements in structures. This also includes transformation of hard rocks to soft soil,
which supports plant growth and is a positive process in nature. However, when this rock is used
as a building block or a constituent of concrete, this biodegradation process is far from positive.
The building materials may be protected by traditional systems such as coatings and hydrophobic
sealers or with organic dispersions. In the bacterial treatment, the solution medium used was of
equimolar concentration of urea (20g/l) and CaCl2 or Ca(NO3)2 for 3 days and thereafter dried
for 3 days at 28oC. The bacteria used were B. Sphaericus(BS). But to protect these bacteria from
the strong alkaline environment in concrete, they were immobilized in Silica Sol-gel. The
treatments were applied by placing the samples on plastic rods in the treatment solution, where
the liquid level was 10 mm above their lower side. Remediation of cracks could be possible by
formation of biocers.

• KanthaD.Arunachalam , K.S. Sathyanarayanan , B.S. Darshan, R.Balaji Raja has published a

article on Biosealant properties of Bacillus sphaericus in which they say Bacillus spharecius was
yet another partially characterized species with similar entity, having the capability of
precipitating calcium carbonate. Earlier researchers have shown very less implementation of the
8 organism in remediation aspect. Bacillus spharecius was sub cultured and temperature, pH
were optimized at 7.4 and 37°C. Growth curve for Bacillus spharecius showed that the log phase
was between 4-11 hours and after 21 hours the bacterial growth was inhibited. EDTA titration
was performed to find out the amount of CaCO3 precipitate and it was highest at pH 8. The broth
culture was subjected to Atomic Force Microscope studies. The analysis confirmed the presence
of calcite in both the bacterial solution and dry scrapes. Optimum nickel ion concentration for
calcium carbonate precipitation was found to be 80μm. The cubes were treated for 5 days in
laboratory scale and to pilot scale in the second phase for 25 days. At the end of the study, the
potential of Bacillus spharecius in Bio-concrete was well established.

• P S Tan, M Q Zhang, D Bhattacharyya has published a article on Processing and Performance

of Self-Healing Materials says Two self-healing methods were implemented into composite
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materials with self healing capabilities, using hollow glass fibres (HGF) and microencapsulated
epoxy resin with mercaptan as the hardener. For the HGF approach, two perpendicular layers of
HGF were put into an E-glass/epoxy composite, and were filled with coloured epoxy resin and
hardener. The HGF samples had a novel ball indentation test method done on them. The samples
were analysed using micro-CT scanning, confocal microscopy and penetrant dye. Micro-CT and
confocal microscopy produced limited success, but their viability was established. Penetrant dye
images showed resin obstructing flow of dye through damage regions, suggesting infiltration of
resin into cracks. Three-point bend tests showed that overall performance could be affected by
the flaws arising from embedding HGF in the material. For the microcapsule approach, samples
were prepared for novel double-torsion tests used to generate large cracks. The samples were
compared with pure resin samples by analysing them using photo elastic imaging and scanning
electron microscope (SEM) on crack surfaces. Further double-torsion testing showed that healing
recovered approximately 24% of material strength. Self-healing materials are materials designed
to recover strength from low-level damage done to the material over the course of its service
lifetime. The self-healing
technique is particularly useful when applied to composite materials, since composites have low
damage detectability and is susceptible to sudden and brittle failure. This study is aimed at two
self-healing methods that had been implemented into composite materials with self-healing
capabilities, that is: (1) using embedded hollow glass fibres (HGF) storing epoxy resin and
hardener, and (2) using microencapsulated epoxy resin with 2- methylimidazole/CuBr2 as the
hardener. Current methods of evaluating the performance of self-healing polymer composites
involve inducing some form of damage into the material.

• SakinaNajmuddinSaifeeet .al1 published a paper on Critical appraisal on Bacterial Concrete.

In this paper they discussed about the different types of bacteria and their applications. The
bacterial concrete is very much useful in increasing the durability of Cementous materials, repair
of limestone monuments, sealing of concrete cracks to highly durable cracks etc. It also useful
for construction of low cost durable roads, high strength buildings with more bearing capacity,
erosion prevention of loose sands and low cost durable houses. They have also briefed about the
working principle of bacterial concrete as a repair material. It was also observed in the study that
the metabolic activities in the microorganisms taking place inside the concrete results into
increasing the overall performance of concrete including its compressive strength. This study
also explains the chemical process to remediate cracks. Bacterial Concrete:

• Meera C M and Dr. Subha V2, have published a paper on Strength and Durability assessment
Of Bacteria Based Self-Healing Concrete. In this paper they have discussed about the effect of
on the strength and durability of concrete. They used cubes of sizes 150mm x 150mm x 150mm
and cylinders with a diameter of 100mm and a height of 200mm with and without addition of
microorganisms, of M20 grade concrete. For strength assessments, cubes were tested for
different bacterial concentrations at 7 days and 28 days and cylinders were tested for split tensile
strength at 28 days. It was observed that the compressive strength of concrete showed significant
increase by 42% for cell concentration of 105 of mixing water. And, with the addition of bacteria
there is a significant increase in the tensile strength by 63% for a bacteria concentration of
105cells/ml at 28 days. For durability assessment, acid durability test, chloride test and water
absorption test were done. From the results it could be inferred that the addition of bacteria
prevents the loss in weight during acid exposure to a certain limit, proving the bacterial concrete
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to have higher Acid Attack Factor. The Water Absorption Test showed a lesser increase in weight
of bacteria concrete sample than control, from which it could be reckoned that the concrete will
become less porous due to the formation of Calcium Carbonate, due to which it resulted in lesser
water absorption rate. Chloride test results showed that the addition of bacteria decreases weight
loss, due to Chloride exposure and enhances the Compressive Strength.

• Ravindranatha, N. Kannan, Likhit M. L3, have published a paper on Self-Healing Material

Bacterial Concrete. In this paper a comparison study was made with concrete cubes and beams
subjected to compressive and flexural strength tests with and without the bacterium Bacillus
pasteurii. The concrete cubes and beams were prepared by adding calculated quantity of bacterial
solution and they were tested for 7 and 28 day compressive and flexural strengths. It was found
that there was high increase in strength and healing of cracks subjected to loading on the concrete
specimens. The microbe proved to be efficient in enhancing the properties of the concrete by
achieving a very high initial strength increase. The calcium carbonate produced by the bacteria
has filled some percentage of void volume thereby making the texture more compact and
resistive to seepage.

• A.T.Manikandan1, A.Padmavathi4, have published a paper on An Experimental Investigation

on Improvement of Concrete Serviceability by using Bacterial Mineral Precipitation. In this
paper, the bacteria Bacillus subtilis strain 121 was from Microbial Type Culture Collection and
Gene Bank, Chandigarh. Samples were prepared in sets of three for a water cement ratio of 0.5
by mass for conventional concrete and a water cement ratio of 0.25 and bacterial culture of 0.25
for bacterial concrete by mass. The cubes were tested by Non-Destructive Testing and HEICO
compression testing machine on the 3rd, 7th and 28th daysafter casting. There was an
improvement in compressive strength by B. subtilis strain 121 due to deposition of Calcite
(CaCO3) in cement-sand matrix of microbial concrete which remediate the pore structure within
the mortar. The temperature sustainability test of B. subtilis in bacterial concrete was carried out
at various temperatures and found that the B. subtilis was found to be alive at –30 C low
temperatures to 700 C high temperatures. There is increase in compressive strength of the
bacterial concrete with B. subtilis bacteria with microbial calcite precipitation in the crack sample
was examined in SEM. The sample showed the presence of calcite crystals grown all over the
surface of the crack and also the presence of B. subtilis bacteria is the evidence, that suggests
microbial remediation properties of bacterial concrete.

• Jagadeesha Kumar B G, R Prabhakara and Pushpa H5, published a paper on Effect of Bacterial
Calcite Precipitation on Compressive Strength of Mortar Cubes. This paper describes about the
experimental investigations carried out on mortar cubes which were subjected to bacterial
precipitation by different bacterial strains and influence of bacterial calcite precipitation on the
compressive strength of mortar cube on 7, 14 and 28 days of bacterial treatment. Three bacterial
strains Bacillus flexus, isolated from concrete environment, Bacillus pasturiiand Bacillus
sphaericus were used. The cubes were immersed in bacterial and culture medium for above
mentioned days with control cubes immersed in water and was tested for compressive strength.
The result indicated that there was an improvement in the compressive strength in the
Pappupreethi K, RajishaVelluvaAmmakunnoth and P. Magudeaswaran early strength of cubes
which were reduced with time. Among the three strains of bacteria, Cubes treated with Bacillus
flexus, which is not reported as bacteria for calcite precipitation has shown maximum
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compressive strength than the other two bacterial strains and control cubes. It was studied that
the increase in compressive strengths is mainly due to consolidation of the pores inside the
cement mortar cubes with micro biologically induced Calcium Carbonate precipitation. The
urease activity was determined for all the bacteria in Urease media by measuring the amount of
ammonia released from urea according to the phenolhypochlorit eassay method. All the three
strains of bacteria were tested for urease activity. The change of the color of the media from
yellow to pink indicated that it is urease positive. All the three strains were urease positive. X-
ray diffraction analysis was also carried out to determine chemical composition of the
precipitation that occurred due to bacterial mineralization.

• RA. B. Depaaand T. Felix Kala6, have published a paper on Experimental Investigation of Self-
Healing Behavior of Concrete using Silica Fume and GGBFS as Mineral Admixtures. In this
paper cubes have been prepared by adding silica fume in percentage of 2.5%, 5%,7.5%, 10%,
12.5% as a binder in addition to adding cement to concrete and by replacing 35% and 55% of
cement with GGBFS.A conventional mixture without any admixture is cast for comparing the
strength and durability properties of silica fume and GGBFS concretes. The specimens are first
tested for compressive strength at 28 days, and then 70% and 90% of the compressive load is
applied to another set of specimens to generate microcracks for studying the durability properties
of the specimens. The preloaded concrete specimens are tested for compressive strength at 7 and
28 days and sorptivity index tests after 28 days. The concrete mix containing cement replaced
with 35% GGBFS has given maximum compressive strength value. Further when silica fume is
added as mineral admixture, the mix has given maximum strength at 12.5% addition of silica
fume.

• Chithra P Bai and Shibi Varghese7, have published a paper on an experimental investigation
on the strength properties of fly ash based Bacterial concrete. In this paper, The bacteria Bacillus
Subtilis was used for study with different cell concentrations of 103, 105 and 107 cells/ml for
preparing the bacterial concrete. Cement was partially replaced by 10%, 20% and 30% of fly ash
by weight for making the bacterial concrete. Concrete of grade M30 was prepared and tests such
as Compressive strength, Split tensile strength, Flexural strength and Ultrasonic Pulse Velocity
were conducted after 28 and 56 days of water curing. For fly ash concrete, maximum
compressive strength, split tensile strength, flexural Strength and Ultrasonic Pulse Velocity
values were obtained for 10% fly ash replacement. For bacterial concrete maximum compressive
strength, split tensile strength, flexural strength, and UPV values were obtained for the bacteria
cell concentration of 105cells/ml. The improvement in the strength properties of fly ash concrete
is due to the precipitation of calcium carbonate (CaCO3) in the micro environment by the bacteria
Bacillus Subtilis.

• V Srinivasa Reddy, M V SeshagiriRaoand S Sushma8, have published a paper on Feasibility

Study on Bacterial Concrete as an innovative self-crack healing system. This paper describes
about the effect of bacterial cell concentration of Bacillus subtilis JC3, on the strength, by
determining the compressive strength of standard cement mortar cubes of different grades,
incorporated with various bacterial cell concentrations. This shows that the Improvement in
compressive strength reaches a maximum at about 105/ml cell concentration. The cost of using
microbial concrete compared to conventional concrete which is critical in determining the
economic feasibility of the technology, is also studied. The cost analysis showed an increase in
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cost of 2.3 to 3.9 times between microbial concrete and conventional concrete with decrease of
grade. And nutrients such as inexpensive, high protein- containing industrial wastes such as corn
steep liquor (CSL)
or lactose mother liquor (LML) effluent from starch industry can also be used, so that overall
process cost reduces dramatically. Precipitation of these crystals inside the gel matrix also
enhances the durability of concrete significantly. Furthermore, this analysis has shown an
increase in the cost of production and a significant decrease in carbon footprint compared to
conventional concrete.

• MohitGoyal and P. Krishna Chaitanya9 published a paper on Behavior of Bacterial Concrete

as Self-Healing Material. In this paper they have carried out laboratory investigations to compare
the different Bacterial Concrete: A Review parameter of bacterial concrete with ordinary
concrete and concrete, in which 70% cement was partially replaced with 30% of Fly Ash and
30% of GGBS. In this paper, Bacillus
pasteurii, is used to prepare M25 concrete. Various tests such as slump flow test, compressive
strength, flexural strength and split tensile strength were conducted for different specimens of,
bacterial concentrations of 40ml, 50ml and 60 ml for each specimen. To identify atomic and
molecular structure and to check the presence of formation of calcium carbonate X- Ray
diffraction test was conducted. There was significant improvement of compressive strength by
30% in concrete mix with bacteria and more than 15% in fly ash and 20% in GGBS. It was
observed that bacterial concrete achieves maximum split tensile strength and flexural strength
when 40 ml and 50 ml bacterial solution was used but loses this trend after 14 days with 60ml
bacterial solution when flexural strength test was performed. Also, 50ml bacterial solution
proved to be effective in increasing the split tensile strength, compressive strength and flexural
strength of the specimen as compared to 40ml and 60 ml bacterial solution. Also, from the XRD
analysis, it is proven that the presence on bacteria is contributing to CaCO3 production, which
has reduced the percentage of air voids, thus, increasing the strength of the structure
considerably.

• N. Ganesh Babu and Dr. S. Siddiraju10,has published a paper on an experimental study on

strength and fracture properties of self-healing concrete. In this paper they have tried is made to
arrest the cracks in concrete using bacteria and calcium lactate. The percentages of bacteria
selected for the study are 3.5% and 5% by weight of cement. In addition, calcium lactate was
used at 5% and 10% replacement of cement by weight. Bacteria produce calcium carbonate
crystals which blocks the micro cracks and pores in the concrete after reacting with calcium
lactate. Bacillus pasteuriiis used for different bacterial concentrations for M40 grade of concrete.
Various tests such as compressive strength, elastic modulus and fracture of concrete were
analyzed. The cubes of dimensions of 100x100x100 mm were used for compressive strength test.
It was observed that compressive strength for controlled concrete using calcium lactate, at 7 days
and 28 days were 19.8 MPa and 40.53 MPa respectively. With the addition of calcium lactate,
there is considerable decrease in compressive strength. Compressive strength of concrete with
5% bacteria was found to be 49.5 Mpa at 28 days, which is more than controlled concrete. With
the addition of calcium lactate at 10% (optimum percentage) and bacteria to concrete, there is
considerable increase in compressive strength. Hence calcium lactate along with 3.5% and 5%
bacteria can be used as an effective self-healing agent.

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CHAPTER - 3
MATERIALS REQUIRED FOR EXPERIMENTAL WORK

3.1 Materials required for experimental study

The ordinary concrete used in the test program consisted of cementing materials, mineral
aggregates and corrosion inhibitor with the following specifications:

 Ordinary Portland Cement

 Flyash
 Graded fine aggregates.
 Graded coarse aggregates.
 Water
 Chemical admixture.
 Bacteria – Bacillus Substills
 Calcium lactate
 Urea Cacl2
a. Ordinary Portland Cement

OPC 53 Grade at is produced Conformed to the Indian Standards Specifications as per IS: 12269-
1987. The cement that is prepared by grinding Portland Cement Clinker and suitable proportions
of Gypsum is called as Ordinary Portland cement. Small additions of performance improvers
such as Fly ash, Blast furnace slag etc. are permitted.
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53 grade Ordinary Portland Cement is high strength OPC and provides numerous advantages
wherever concrete for special high strength applications is required. The rate of development of
strength is faster than 43 grade OPC.
53 Grade OPC is different from Rapid Hardening Portland Cement IS: 8041and their purposes
of usage are different.
The Cement mortar that is prepared in 1: 3 proportions using standard sand (as per IS:650) and
tested at standard laboratory conditions and gains the compressive strength of not less than 53
MPa after 28 days, then the Cement is said to be 53 Grade Ordinary Portland Cement.

b. Flyash:
Fly ash is a byproduct from burning pulverized coal in electric power generating plants. During
combustion, mineral impurities in the coal (clay, feldspar, quartz, and shale) fuse in suspension
and float out of the combustion chamber with the exhaust gases. As the fused material rises, it
cools and solidifies into spherical glassy particles called fly ash. Fly ash is collected from the
exhaust gases by electrostatic precipitators or bag filters. The fine powder does resemble Portland
cement but it is chemically different. Fly ash chemically reacts with the byproduct calcium
hydroxide released by the chemical reaction between cement and water to form additional
cementitious products that improve many desirable properties of concrete. All fly ashes exhibit
cementitious properties to varying degrees depending on the chemical and physical properties of
both the fly ash and cement. Compared to cement and water, the chemical reaction between fly
ash and calcium hydroxide typically is slower resulting in delayed hardening of the concrete.
Delayed concrete hardening coupled with the variability of fly ash properties can create
significant challenges for the concrete producer and finisher when placing steel-troweled floors.

c. Graded Fine Aggregates

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The materials smaller than 4.75 mm size is called fine aggregates. Natural sand is generally
used as fine aggregate. In this experimental work replacement of river sand by quarry waste
(fineness modulus of crushed sand equal to 3.2) conforming to grading Zone III of IS – 383 –
1970 was used as fine aggregates.

d. Graded Coarse Aggregate

Locally available well graded granite aggregates of normal size greater than 4.75 mm and less
than 16mm having fineness modulus of 2.72 was used as coarse aggregates.

e. Water
Potable water has been used for casting concrete specimens. The water is free from oils, acids,
and alkalis and has a water-soluble Chloride content of 140 mg/lit. As per IS 456 – 2000, the
permissible limit for chloride is 500 mg/lit for reinforced concrete; hence the amount of
chloride present is very less than the permissible limit.

f. Chemical admixture
Chemical admixtures are the ingredients in concrete other than portland cement, water, and
aggregate that are added to the mix immediately before or during mixing. Producers use
admixtures primarily to reduce the cost of concrete construction; to modify the properties of
hardened concrete; to ensure the quality of concrete during mixing, transporting, placing, and
curing; and to overcome certain emergencies during concrete operations.

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g. Bacteria
Bacillus substilis is an obligate aerobe bacterium used as a larvicide for mosquito control. It
forms spherical endospores. Bacillus substilis is a gram-positive bacterium, with rod shaped cells
that form chains-Medium-sized, smooth colonies with an entire margin. and Rod-shaped cells.
Gram-variable, large, spore-forming rods with a diameter < 0.9 μm. Catalase -positive.
Lecithinase-negative. Does not attack sugars. Grow thing range of Temperature: 37oc Optimum
Temperature- 35-37^oC

3.2. Tests on Material

3.2.1. Specific Gravity and Water Absorption

Specific Gravity helps in measuring the quality of aggregate used. It is defined as the ratio of
mass of any substance to the mass of equivalent volume of water. Aggregates having lower
specific gravity are considered as weak than the aggregates having higher specific gravity. If the
water absorption value of aggregates is high, then they are weak and porous. It is determined
as per IS: 2386 (Part III) –1963.
In case of coarse aggregates, the specific gravity is obtained by using wire basket. About 2kg of
coarse aggregates are tested. The aggregates are kept in the wire basket and submerged in
water. Air entrapped on the surface of aggregate shall be expelled by gentle disturbance or by
rapid clockwise and anti-clockwise movement of wire basket. The basket and aggregate remain
submerged in water for 24 hrs. Then the aggregates are surface dried and weighed. After that
the aggregates are oven dried.

Specific gravity = W4 W3−(W1−W2)

Apparent Specific gravity = W4 W4−(W1−W2)
Water absorption = W3−W4 W4 × 100
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Where,
W1 = weight of wire basket containing sample and filled with distilled water, gm
W2 = weight of wire basket filled with distilled water only, gm
W3 = weight of saturated and surface-dry aggregate, gm
W4 = weight of oven-dry aggregate, gm

In case of fine aggregate pycnometer is used for determination specific gravity as per IS: 2386
(Part III) –1963. Sample of weight 500gm is taken for test. Saturated surface dry aggregates are
used for the testing. These aggregates are then deposited in pycnometer and distilled water is
filled to the top so that water in the hole is flat and its weight is taken. Weight of pycnometer
is taken when it is filled with water. Then fine aggregate is oven dried.

Specific gravity = W4 W1−(W2−W3)

Apparent specific gravity = W4 W4−(W2−W3)
Water absorption = (W1−W4) W4 × 100

Where,
W1 = weight of saturated and surface-dry fine aggregate, gm
W2 = weight of pycnometer containing fine aggregate and filled with distilled water, gm
W3 = weight of pycnometer filled with distilled water, gm
W4 = weight of oven-dried fine aggregate, gm
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Specific gravity of cement

In case of cementitious material specific gravity is obtained by using Le-Ch atelier flask.
Kerosene oil or Naphtha is filled in the flask in between 0- and 1-ml. Specific gravity of
cementitious material is determined as per IS: 4031 (part XI)- 1988.

Le-Ch atelier Flask Initial reading is noted down. Using funnel cementitious material is filled in
the flask of about 60 gm.
The final reading is taken. Specific gravity is given by, G = Mass of cement, gm Displaced volume,
cm 3

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3.2.2. Aggregate Crushing Test

Aggregate crushing test is carried out to determine the strength of aggregate. It is determined
as per the IS: 2386 (Part IV) -1963. Surface dry aggregate is used which passes through 12.5 mm
IS sieve and retain on IS sieve 10 mm is filled in three equal layers in a mould of cylindrical
shape, each layer being ramped 25 times by the tamper. The plunger is placed on the top of
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specimen and a load of 40 tones is applied at certain rate by the compression machine. The
crushed aggregates are sieved through 2.36 mm IS sieve. Strong aggregate give low aggregate
crushing value.

Aggregate crushing value = W2 W1 × 100 percent (3.8)

Where W1 = weight of surface dry aggregate
W2 = weight of crushed aggregate passing 2.36 mm IS sieve.

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3.2.3. Aggregate Impact Test

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This test carried out to determine the toughness or resistance of aggregate to fracture under
repeated impacts. It is determined as per IS: 2386 (Part IV) -1963.
Impact Testing Machine The aggregates which passes 12.5mm sieve and retain on 10mm sieve
is filled in a mould of inner diameter 10.2cm and depth 5cm in three layers and giving each layer
25 blows. The hammer of weight 13.5-14 kg is lifted to a ht. of 380mm above the top surface
of mould in which aggregates are placed and allowed to drop on the specimen. The aggregates
are subjected to 15 blows with 1 sec interval. The crushed aggregates are sieved through
2.36mm.

Impact value = W2 W1 × 100 percent

Where W1 = weight of surface dry aggregate
W2 = weight of crushed aggregate passing 2.36 mm sieve.

3.2.4. Aggregate Abrasion Test

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This test is carried out to determine the hardness of aggregates. The test is carried out as per
IS: 2386 (Part IV) -1963. Los Angeles abrasion testing machine is used for the testing which is a
hollow steel cylinder closed at both ends and having internal diameter of 700mm and length of
500mm. A steel shelf is radially projected 88mm for the full length of cylinder. Specified weight
of aggregate depending upon the gradation is placed in the machine. The machine rotates at a
speed of 33rpm for specified number of rotations as per grading. Then the aggregates are taken
and sieved through 1.7mm sieve.

Los Angeles Abrasion Testing Machine Abrasion value = W2 W1 × 100 (3.10)

Where W1 = weight of surface dry aggregate

W2 = weight of crushed aggregate passing 1.7 mm sieve.

3.2.5. Consistency of Cementitious Material

Consistency is the percentage of water required for cement paste at which viscosity of the paste
becomes such that the plunger in a Vicat's apparatus penetrates a depth of 5 to 7mm, measured
21 from the bottom of Vicat mould. Consistency of cementitious material is determined as per
IS: 4031 (Part IV) – 1988. In this test measured quantity of cementitious material is mixed with
measured quantity of potable or distilled water, care should be taken such that the gauging
time should not be less than 3 minutes and not more than 5 minutes. The gauging time is the
time of mixing water to dry cementitious material up to the commencing of filling the mould.
The Vicat mould is kept on non-porous plate. Mould is filled with cement paste and leveled
using trowel.
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Vicat's Apparatus Mould is slightly shaken to expel air. Plunger is attached to the apparatus and
allowed to rest on the surface of the test mould. Then the plunger is quickly released to sink
into the mould. This procedure is repeated until plunger penetrates 5 to 7 mm from the bottom
by adjusting the quantity of water added.

Consistency = A B × 100 = P
Where A = quantity of water added
B = quantity of cementitious material used

3.2.6. Soundness

Test is carried out to detect the presence of uncombined lime in cement. It is the property by
virtue of which the cement does not undergo any appreciable expansion (or change in volume)
after it has set, thus eliminating any chances of disrupting the mortar or concrete. The
apparatus used for soundness test is Le-chatelier apparatus. Soundness test is carried out as
per IS: 4031 (Part III) - 1988. The mould and glass sheets are lightly oiled and cement paste
formed by adding cement with 0.78 times the water required to form paste of standard
consistency is placed in the Le chatelier’s mould by placing a glass sheet below and holding the
two edges together.
Le-chatelier Apparatus
The Le-chatelier mould is covered with other glass sheet and a weight is placed over the whole
assembly. Immediately submerge the whole assembly in water for 24 hours. The distance
between the two indicators is measured. Again, immerse the sample in water and boil for 3
hours and distance between both indicators are noted. The difference of both the reading
indicates the expansion of cement.

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3.2.7. Initial and Final Setting Time

Initial setting time is the time period that elapses from the time when water is added to the
cementitious material and the needle for initial setting time ceases to penetrate 5 to 7 mm
from bottom of the Vicat’s mould.

Vicat’s Apparatus Final setting time is the time period that elapses from the moment water is
added to the cementitious material and the needle for final setting time with annular collar at
the tip of needle just makes an impression on the paste. Initial and final setting time of
cementitious material is determined as per IS: 4031(PART V) – 1988. For this test measured
quantity of cementitious material is taken and mixed with 0.85times the water required to form
standard consistency paste. Gauging time is maintained. Needle is used for initial setting time
and for final setting time needle with annular attachment is used.

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3.2.8. Fineness

The fineness of cement is a measure of cement particle size and is denoted as terms of the
specific surface area of cement. The Fineness Test of Cement is done by sieving cement sample
through standard IS sieve. The weight of cement particle whose size is greater than 90 microns is

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determined and the percentage of retained cement particle are calculated. This is known as
the Fineness of cement.
Equipment’s required to check fineness of cement
 90µm IS Sieve,
 Weight Balance having capacity 10 mg to 100 g,
 Nylon or pure bristle brush
Fineness Test of Cement Procedure

1. Collect a sample of cement and rub with your hands. The Fineness test sample should be free
of lumps.
2. Take 100 gm of cement sample and note its weight as W1.
3. Drop 100 gm of cement in 90 µm sieve and close it with the lid.
4. Now, shake the sieve with your hands by agitating the sieve
in planetary and linear movements for 15 minutes.
5. After that take weight the retained cement on the 90 µm sieve as W2. To calculate fineness of
cement formula is given below

Fineness = (W2/W1) * 100

 Then, calculate the percentage of Weight of cement–retained on Sieve.

 Repeat this procedure with three different samples of cement and average the values for
accurate results.

3.2.9. Flakiness and Elongation Index

The flakiness and elongation index are determined as per IS: 2386 (Part IV) -1963. The flakiness
index of an aggregate is the percentage by weight of particles in it whose least dimension
(thickness) is less than three-fifths of their mean dimension. The test is not applicable to sizes
smaller than 6.3 mm. The thickness gauge is used for flakiness of aggregate.

The measured quantity of material is sieved through the sieve size mentioned in the metal
gauge and collected separately as per range. Then the fraction of material is gauged through
thickness in metal gauge. The total mass of each size fraction of the sample also shall be
determined. The mass of material passing the respective gauge to the total mass of aggregate
retained on 6.3mm sieve gives the flakiness index of aggregate which is expressed in
percentage. Elongation index of an aggregate is the percentage by weight of particles whose
greatest dimension (length) is greater than one and four-fifths times their mean dimension.
Normally, the properties of interest to the engineer are sufficiently covered by the flakiness or
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angularity tests. The elongation test is not applicable to sizes smaller than 6.3 mm. The fraction
of material is gauged through length in metal gauge. The total weight of material retained in
every range to the total weight of material retained.

3.2.10. Sieve Analysis of Aggregates

This method is adopted to determine the particle size distribution of fine and coarse aggregate.
It is carried out as per IS: 2386 (Part I) – 1963. Set of sieves are used for analysis of both fine
and coarse aggregates which are arranged in descending order. Measured quantity of air dry
26 aggregates are used. Aggregates are passed through the set of sieves and material retained
on each sieve is weighed. The result is calculated as cumulative percentage by weight of the
total sample passing each of the sieves, to the nearest whole number. The result is represented
graphically.

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3.3 Mix Proportioning

The quality of concrete mainly depends upon the proportioning of its constituent’s materials. The
mix proportioning mainly influences the permeability of concrete and cause cracks. Design mix
is more appropriate to know the exact behavior of cracks rather than nominal mix. In this
experiment the concrete used is 25 MPa having water to cement ratio of 0.42.

3.4 Mix Design

The aim of studying the various properties of materials of concrete, plastic concrete and hardened
concrete is to design a concrete mix for strength. Design of concrete mix needs complete
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knowledge of the various properties of the constituent material, the implications in place of
change on the conditions at site, the impact of the properties of plastic concrete on the hardened
concrete and the complicated interrelationship between the variables. Mix design can be defined
as the process of selecting suitable ingredients of concrete and determining their relative
proportions with the object of producing concrete of certain minimum strength and durability as
economically as possible. The mix design procedure is explained in the following section.

3.5 Indian Standard Mix Design Stipulations

a. Concrete grade : M25

b. Exposure : Moderate
c. Quality control : Fair
d. Size of aggregate : 20 mm
e. Degree of workability : Very good
f. Cement used : OPC 53 grade cement
g. Sand grading zone : II
h. Minimum Cementitious content : 300 Kg/cum, as per IS-456:2000,
Clause No.: 8.2.5,
i. Maximum Cement (OPC) content : 450 Kg/cum (As per IS: 456 :2000,
Clause no.8.2.4
ratio : 0.50 (As per IS-456:2000, Clause No.: 8.2.5,
Table – 5
k. Workability (Slump) : 75-150mm
l. Degree of supervision : Very good
m. Type of coarse aggregate : Single Sized Crushed Angular Aggregate of
Nominal Size 20mm (As per IS 383: 2016)
n. Type of fine aggregate : River sand conforming IS 383:2016
o. Chemical admixture : Superplasticizer conforming to IS :9103

3.6 Test data for raw material

1 Cement used :- ULTRATECH OPC 53

2 Specific Gravity of Cement :- 3.15
Source of Admixture
3 :- CHRYSO OPTIMA S857
Used
4 Specific Gravity of Admixture :- 1.12

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5 Specific Gravity of Aggregate

a) Coarse Aggregate
i) 20mm Agg. :- 2.614
ii) 10mm Agg. :- 2.603
b) Fine Aggregate :- 2.588
6 Water Absorption of Aggregate %
a) Coarse Aggregate
i) 20mm Agg. :- 0.56
ii) 10mm Agg. :- 0.84
b) Fine Aggregate :- 1.32
7 Free (Surface) Moisture
a) Coarse Aggregate
i) 20mm Agg. :- Nil
ii) 10mm Agg. :- Nil
b) Fine Aggregate :- Nil
8 Combined EI + FI of Aggregate :- 18.40%
9 Aggregate Impact Value :- 16.81%
Sieve analysis
10.
1) Coarse Aggregate:-

Coarse Aggregate
Percentage of Different Fractions
IS Sieve Fraction Limits As
Sizes per IS : 383
10
(mm) 20 mm (Table -7)
20 mm 10 mm mm Combined
0 50
40 100 100 50 50 100.0 100
20 92.42 100 46.21 50.00 96.21 90-100
10 5.52 92.44 2.76 46.22 48.98 25-55
4.75 0.92 5.43 0.46 2.72 3.18 0-10

2) Fine Aggregate: - Conforming to Grading Zone II of Table 9 of IS 383

Table 3.1: Mix design summary

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Material Weight(kg) Volume(m3)

Cement 255 0.081

Fly ash 85 0.039

Coarse Aggregate 570 0.218

(20MM)
Coarse Aggregate 569 0.218
(10MM)
Fine Aggregate 778 0.300

Water 142 0.142

Admixture 2.04 0.002

Total 2401 1.000

CHAPTER - 4
PREPARATION OF BACTERIA

4.1 Mixing of Bacteria

Luria Berta-powder form (6.75gms) +500ml of distilled water peptone(3gms)+yeast
extract(1.5gms)+Beef extract(1.5gms) +sodium chloride (3gms/100ml) +1Loop of Bacteria (gel
medium) = Incubator 37oC

4.2 Preparation of Nutrient Agar

Bacteriological media came a wide range of types. Nutrient Agar is a complex medium. Nutrient
Agar contains beef extract (0.3%), peptone (0.5%), and Agar (1.5%) in water. Beef extract is
prepared as dehydrated form of autolyzed beef and is supplied in the form of paste. Peptone is
casein (milk protein) that has been digested with the enzyme pepsin. Peptone is dehydrated and
supplied as a powder. peptone and Beef extract contains a mixture of amino acids and peptides.
Beef extract also contains water soluble digest products of all other macro molecules (nucleic
acids, fats, polysaccharides) as well as vitamins trace minerals. Agar is purified from red algae
in which it is an accessory polysaccharide (polyelectronic acid) of their cell walls. Agar is added
to microbiological media only as a solidification agent. Agar for most purposes has no nutrient
value. Agar is an excellent solidification agent because it dissolves at near boiling point solidifier
at 45ºC. Thus, one can prepare molten (liquid) agar at 45ºC, mix cells with it, then allow it to
solidify thereby trapping living cells. Below 45ºC agar is a solid and remains so as the
temperature is raised melting only when greater than 95oC is obtained.
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In this method Bacteria are added during casting of concrete. The amount of Bacteria added
in the range of 10ml & 15 ml/m3 of concrete. Concrete could soon be healing its own hairline
cracking. Holes and pores of wet concrete are healed. Combined calcium with oxygen and
carbon di oxide to form calcite is essential for healing tiny cracks which arrest the seepage of
water. Figure 4.4.1: View on Growthing of Bacteria
The technique of using soil bacterium is highly desirable because the mineral precipitation
induced because of microbial activities, is pollution free and natural. Bacillus spharecius was yet
another partially characterized species, having the capability of precipitating calcium carbonate.
Its Far better would be to use bacillus sphaericus as a material that heals itself just as the cell
divines and produces a visible mass. The colony isolated from other colonies; isolated colonies
are assumed to be pure culture.

4.3 Culturing and Isolation

Microorganism must have a constant nutrient supply if they are to survive. Media may be liquid
(broth) or solid (agar). Any desired nutrients may be in corporate into the broth (or) agar to grow
bacteria. Organism grown in broth cultures causes turbidity, (or) cloudiness, in the broth. On
agar, masses of cells known as colonies, appear after a period of incusation certain separated on
agar so that as the cell divines and produces a visible mass. The colony isolated from other
colonies; isolated colonies are assumed to be pure culture. A. Gandhimathi, N.
Vigneswari, S.M. Janani, D. Ramya, D. Suji and T. Meenambal 22 ISBN 978-93-82338-32-1| ©
2012 Bonfring

4.4 Ability of Bacterial Concrete to Repair the Cracks

Both attentions will be given on closure of cracks (blocking the path for ingress of water and
ions) and on regaining mechanical properties. Cracks in concrete specimen subjected to various
loading situations will be investigated before and after the healing. For this impregnation
techniques and SEM will be applied. (Scanning electron microscope). On the other hand, the
micro-organisms such as bacteria, cyono bacteria, algae, lichens, yeasts, fungi and mosses etc.
Which are omnipresent and omnipotent are responsible for metabolism action that results in a
microbial deposition of a protective CaCO3 layer. Also, this process results in re-establishment
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of the cohesion b\n particles of mineral building materials and protects against further decay of
stone material. To prove the positive effects of microbial CaCO3 precipitation. The increase in
porosity in concrete leads to increase in capillary water uptake, increase in gas permeability
along with higher carbonation rate, high chloride migration and freeze-thaw damage.

4.5 Processing of Bacteria

Concrete could soon be healing its own hairline cracking. Holes and pores of wet concrete are
healed. Combined calcium with oxygen and carbon di oxide to form calcite is essential for
healing tiny cracks which arrest the seepage of water

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CHAPTER – 5

EXPERIMENTAL STUDY

5.1 Process of manufacture of concrete

Production of quality concrete requires meticulous care exercised at every stage of manufacture
of concrete. If meticulous care is not exercised, and good rules are not observed, the resultant
concrete is going to be of bad concrete. Therefore, it is necessary for us to know what the good
rules are to be followed in each stage of manufacture of concrete for producing good quality
concrete. The various stages of manufacture of concrete are:
1. Batching
2. Mixing
3. Placing
4. Compacting
5. Curing

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5.1.1 Weigh Batching

Weigh batching is the correct method of measuring the materials. For important concrete,
invariably, weigh batching system should be adopted. Use of weight system in batching,
facilitates accuracy , flexibility and simplicity. Different types of weigh batchers are available,
the particular type to be used, depends upon the nature of job. When weigh batching is adopted,
the measurement of water must be done accurately using measuring jars.

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5.1.2 Electrical portable Mixer

The concrete mixer It is a machine used for the manufacture of mortar and concrete, after
mixing different components such as aggregates of different sizes, water, and cement, basically.
It is a machinery specially designed for the field of construction. It is made up of a body and a
cylindrical container that rotates with great force transmitted by an electric motor.

The electric concrete mixers They have the controls in the form of a button or push button. It is
necessary to take care of its installation to avoid further damage or injury, thus avoiding
accidental activation of the start-up switches and easy operation of the stop buttons. The push
buttons should not be next to the engine, but preferably on the outside, in an easily accessible
place, away from the transmission belt from the engine to the cylinder. It will only be possible
to place the start-up switch next to the transmission belt if it is fully protected.

5.1.3 Placing
It is not enough that a concrete mix correctly designed, batched, mixed, it is of utmost
importance that the concrete must be placed in systematic manner to yield optimum results. The
precautions to be taken and methods adopted while placing concrete in the moulds.

5.1.4. Hand Compaction

Hand compaction of concrete is adopted in case of small concrete works. Sometimes,
t h i s method is also applied in such situation, where a large quantity of reinforcement is used,
which cannot be normally compacted by mechanical means. Hand compaction consists of
rodding, ramming, or tamping. When hand compaction is adopted, the consistency of concrete
is maintained at a high level. Tamping is one of the usual methods adopted in compacting roof
or floor slab or road pavements where the thickness of concrete is comparatively less and
the surface to be finished smooth and level.

5.1.5 Curing
Concrete derives its strength by the hydration of cement particles. The hydration of cement is
not a momentary action but a process continuing for long time. Curing can also be described as
keeping the concrete moist and warm enough so that the hydration of cement can continue. More
elaborately, it can be described as the process of maintaining a satisfactory moisture content and
a favorable temperature in concrete during the period immediately following placement, so that
the hydration of cement may continue until the desired properties are developed to a sufficient
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degree to meet the requirement of service. The casted cubes and cylinders are immersed in water
tanks for 3 days, 7days, 14 days and 28 days.

5.1.6 Workability of Concrete

Workability is the amount of useful internal work required to produce full compaction of
concrete. It depends on,
1. Types of aggregate
2. Grading of coarse and fine aggregate
3. Quantity of cement paste
4. Consistency of the cement paste

5.1.7 Slump Test

Slump test is the most used method of measuring consistency of concrete which can be employed
either in laboratory or at site of work. It is not a suitable method for very wet or very dry concrete.
It is used conveniently as a control test and gives an indication of the uniformity of concrete from
batch to batch. The deformation shows the characteristics of concrete with respect for segregation
The thickness of the metallic sheet for the mould should not be thinner than 1.6mm. for tamping
the concrete, a steel tamping rod 16mm dia, 0.6 meter along with bullet end is used. The mould
is then filled in four layers, each approximately 1/3rd of the mould (As per IS 1199). Each layer
is tamped 25 times by the tamping rod taking care to distribute the strokes evenly over the cross
section. After the top layer has been rodded, the concrete is struck off level with a trowel and
tamping rod. The mould is removed from the concrete immediately by rising it slowly and
carefully in a vertical direction. This allows the concrete to subside. This subside is referred as
slump of concrete.
The value of slump = 90 mm

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5.2 Mixing, Compaction and Curing

Good concrete can be obtained only through and uniform mixing, better through and uniform
mixing, better through compaction and adequate curing. In the laboratory, the concrete was
mixed by electrical portable mixer. All the constituent materials were weighed, and dry mixing
was carried out for about 5 minutes and then water was added. The mixing was continued till
concrete of uniform consistency was obtained / the specimens were compacted using tamping
rod in 3 layers (each layer 35 blows). After 24 hours, the specimens were remolded and kept
immersed curing tank containing potable water till the required curing period is over. The mix
proportions are given in table. For control specimen the w/c ratio is 0.42. The same amount of
water is used for all other specimens.

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CHAPTER – 6
RESULT AND CONCLUSION

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6.1 Result and conclusion

Concrete plays a major role in the construction industry. For a durable structure, good quality
concrete must be used. A Self-Healing Concrete for the Future which says a common soil
bacterium was used to induce calcite precipitation which is highly desirable because the mineral
precipitation induced because of microbial activities is pollution free and natural. The
workability test of the bacterial concrete resulted in 90mm of slump value.
We have found out that the compressive strength of the bacterial concrete with 5% bacteria
+0.005 mol/lit. Calcium lactate+ Urea CaCl2 is maximum in comparison of other compositions

Bacteria is added with the following:

1. Urea CaCl2
2. Buffer -solution (phosphate buffer)

Bacteria will not survive in water. So, it cannot be mixed with water, and it was found out in the
Research when the bacteria mixed with Buffer - solution give better results. Even it will not
change the pH value when added with acid (or) alkali is added to it. The bacteria will be mixed
in different ratios in the specimen concretes for testing and Research. The cost of bacterial
concrete when compared to conventional concrete is more or less the same which will not require
any rehabilitation work which is costlier for rectification of crack after 15 years, but this self-
healing concrete will help in regaining of strength and healing of cracks automatically without
any human intervention
6.2 Compressive Strength Test

The compressive strength had considerable improvement with concrete specimens containing
bacteria and calcium lactate shown in detail given below. The highest increment of compressive
strength for both bacteria is with calcium lactate of 0.005 mol/l concentration. The increment of
concrete specimen strength with Ent.f and Ent.f with calcium lactate of 0.001 mol/l, 0.005 mol/l
and 0.01 mol/l concentration compared to control are 6.11%, 6.9%, 18.9% and 8.3% respectively.
Whereas, the increment of strength for concrete specimens containing B.sp and B.sp with
calcium lactate of 0.001 mol/l, 0.005 mol/l and 0.01 mol/l concentrations compared to control
are 2.7%, 6.9%, 10% and 5% respectively.

Irwan et al., ha started those materials added into concrete which does not participate in hydration
process with the binder could attribute to decrease in compressive strength. Bacteria added into
concrete does not directly involved in the hydration process itself but used the by-product of the
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reaction to produce calcium carbonate. The slight reduction of compressive strength with higher
concentration of calcium lactate shows that over production of calcium carbonate would affect
the compressive strength. Similar result is reported by Faiz and Steve, which stated that minimal
amount of calcium carbonate was found most effective in increment of strength. Researchers have
studied that certain concentration of bacteria increases compressive strength compared to control.

However, the concentration of bacteria depends on the bacteria itself. As every bacterium used
are of different origins. Thus, may have different reactions. In Xuet al. and Abo- El-Enein studies,
calcium source were added into bio concrete. After which compressive strength were tested and
compared. It was found that different calcium source precipitates different amount of calcium
carbonate. Different calcium source influences the rate of bacteria precipitation and amount.

The precipitation of calcium carbonate that fills the pore within the concrete matrix is the factor
for increment of compressive strength. Addition of Ent.fin concrete has
considerable impact on the compressive strength compared to B.sp. The obvious difference for
both bacteria is with 0.005 mol/l concentration of calcium lactate. Ent.fhas shown a tremendous
increase of compressive strength compared to B.sp. This is due to Ent.freaction to the addition of
calcium source. The addition of calcium source allows the bacteria to increase precipitation
which benefited the concrete. Calcium lactate with concentration of 0.01 mol/l has caused an
over production of calcium carbonate which cause the strength to reduce slightly. This is similar
for both bacteria. The increment of strength with addition of bacteria are due to deposition of
calcite facilitated by calcium nutrient.

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Table 6.1

Type of concrete Compressive strength of concrete after 7 days

Sample 1 Sample 2 Sample 3
Conventional 22.7 23.0 23.5
5% Bacillus sp 24.0 24.5 24.8
(B.sp)
5% B.sp+ 0.005 CL 25.7 26.3 26.5
mol/l
5% B.sp+ 0.005 CL 28 28.5 29.4
mol/l+ Urea CaCl2

Table 6.2

Type of concrete Compressive strength of concrete after 14days

Sample 1 Sample 2 Sample 3
Conventional 28.3 29.1 29.5
5% Bacillus sp 29.1 29.9 30.3
(B.sp)
5% B.sp+ 0.005 CL 31.0 30.8 31.9
mol/l
5% B.sp+ 0.005 CL 32.5 32.0 33.3
mol/l + Urea CaCl2

Table 6.3

Type of concrete Compressive strength of concrete after 28 days

Sample 1 Sample 2 Sample 3
Conventional 31.7 32.5 32.9

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5% Bacillus sp 33.2 34.5 34.9

(B.sp)
5% B.sp+ 0.005 CL 34.5 35.8 37.0
mol/l
5% B.sp+ 0.005 CL 35.4 36.5 37.5
mol/l + Urea CaCl2

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30

25

20

15
Sample 1

10 Sample 2

Sample 3

Conventional 5% Bacillus Substils 5% Bacillus 5% Bacillus

Substils+0.005CL mol/l Substils+0.005CL

mol/l+Urea Cacl2

Figure 6.1(Compressive Strength After 7 Days of Curing)

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Sample 1

Sample 2
Sample 3

Conventional 5% Bacillus substils 5% B.sp+ 0.005 CL 5% B.sp+ 0.005 mol/l CL+

mol/l Urea CaCl2

Figure 6.2(Compressive Strength After 14 Days of Curing)

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45

40

35

30

Sample 1
25
Sample 2

Sample 3
20

10

Conventional 5% Bacillus sp 5% B.sp+ 0.005 CL 5% B.sp+ 0.005 CL

(B.sp) mol/l mol/l + Urea CaCl2

Figure 6.3 Compressive Strength After 28 Days of Curing

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6.3 Test on bacteria

Healing agent(i.e, bacteria + calcium lactate) isapplied on the cracked surface

Day 1 healing Agent Applied

After 10 days Gentle curing results in

filling up the crack

20th day photo- healing of small portion of crack completely

58

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Another experiment was conducted in which 2ml of

bacteria was mixed with 2mg of calcium lactate. The
mixture was shaked in test tube and placed in lab for 24hrs
and then sealed for next 24 hours. This result inhardened
mass in tube which confirm formation of calcium
carbonate in tube.

REFERENCES

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