Int. J. Life. Sci. Scienti. Res.

eISSN: 2455-1716
Jokyani and Chouhan, 2018
DOI:10.21276/ijlssr.2018.4.5.10

Research Article

Isolation, Characterization, and Application of Calcite Producing

Bacteria for Self-Healing Concrete Preparation
1 2
Diksha H. Jokyani , Dharmvir Chouhan *
1
Student, P. G Department of Microbiology, Dhote Bandhu Science College, Gondia, India
2
Assistant Professor cum HOD, P. G Department of Microbiology, Dhote Bandhu Science College, Gondia, India

*Address for Correspondence: Prof. D. A. Chouhan, Assistant Professor cum HOD, P.G Department of Microbiology,
Dhote Bandhu Science College, Kudwa Road Gondia, Maharashtra- 441614, India
Received: 18 Apr 2018/ Revised: 05 Jun 2018/ Accepted: 02 Aug 2018

ABSTRACT
Concrete being the foremost building material broadly used in the construction sector is subjected to crack formation due to low
tensile strength, durability, and ductility. So this issue is of great curiosity to the researchers in pursuit for the concrete production
with better properties. Micro-cracks are the main reason for structural failure occurs, when the load applied exceeds its limits.
This causes the seepage of water and other salts. In order to overcome this, the carbonate precipitating, non-pathogenic, spore-
forming, alkaline resistant strain of Bacillus subtilis has been explored as a bio-cementing material. Cracks in M20 grade concrete
4 5 6
blocks are injected by direct means with screened bacterial strain Bacillus subtilis (BH3) at the cell concentration of 10 , 10 , 10
CFU/ml with silica gel as an immobilizing agent and calcium lactate as a food source. The cracks were allowed to heal for
appropriate time duration at specific pH, temperature, and urea concentration. These findings suggest the potential of Bacillus
subtilis in an autogenously healing process.

Key-words: Bacillus subtilis, BH3, Carbonate precipitation, Calcite, Self healing and Bio-cementation

INTRODUCTION
A large amount of non-renewable resource is consumed
by the construction engineering sector, most of which
contribute to the highest proportion of global CO2
emission at their production or application stage. The
concrete production process is an energy-intensive
process if in case of mining, transportation and
processing are considered. Its production level lies at
about 2.35 billion metric tonnes per year and contributes
an astonishing 10% of CO2 emission in the atmosphere [1].
Another issue concerns is the huge maintenance costs
for structure built in past. About 10% of the bridges in
the USA are considered structurally deficient and 10% of
it is considered functionally obsolete [2]. Apart from this
the factors like freeze-thaw reactions, shrinkage, results
How to cite this article
Jokyani DH, Chouhan D. Isolation, Characterization, and
Application of Calcite Producing Bacteria for Self-Healing Concrete
Preparation. Int. J. Life. Sci. Scienti. Res., 2018; 4(5): 2025-2036.
Access this article online
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in the cracking of concrete structures during its
hardening and this ultimately leads to structural
deformation. Latter the ingress of some hazardous
chemicals and moisture into the concrete may lead to
decrement in its serviceability whereas; the penetration
of sulphates and chlorides in cracks causes the durability
to be affected. Hence the more concern should be given
to this so as to prevent the expansion of cracks by a
sustainable means that might involve the natural
microbial mechanism of bio-cementation which is
promising.
Autonomously healed concrete is nothing but the
biologically produced limestone by which the cracks can
be healed effectively. The screened bacterial strain of
genus “Bacillus” along with the calcium based nutrient
like calcium lactate and an immobilizing agent like silica
gel is mixed in appropriate proportion and injected into
the preformed cracks by direct means. In case if it is
added initially to the concrete mixture then the
self-healing agent can lie in the dormant stage within the
concrete for up to about 200 years. Due to any of the
damage the water starts to percolate deep into the
structure and the dormant bacterial form if get the

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Int. J. Life. Sci. Scienti. Res.
growth favourable conditions to acquire its vegetative
form and starts to feed on calcium lactate consuming
oxygen and thereby converting soluble calcium lactate to
insoluble calcium carbonate better called limestone.
Here the consumption of oxygen by the bacterial strain is
advantageous as oxygen is responsible for steel
corrosion thereby enhancing the durability of steel
structures [3], Since 1980’s several researchers had been
working on this concept and had used the different
bacterial strains such as Jonker et al. [4] used Bacillus
cohnii bacteria to precipitate CaCO3 while Santhosh et al.
[5]
and Bang et al. [6] used Bacillus pasteurii.
As the bacterial cell wall is negatively charged, it will
draw the cations from the environment, Ca2+ and deposit
it on their cell surface serving as a nucleation site [6].
Ca2+ + Cell Cell-Ca2+
This bacterial genus is known to have specific urease
activity that is utilized for the hydrolysis of urea into 1
mole of ammonia and 1 mole of carbonic acid [7].
H2N-CO-NH2 + H2O NH3 + H2N-CO-OH
H2N-CO-OH + H2O NH3 + H2CO3 3
These two products was subsequently form 2 moles of
ammonium and hydroxide ion as,
H2CO3 HCO3- + H+
2NH3 + 2H2O 2NH4+ + 2OH-
Reaction 4 and 5 turned in results in pH increase and
shifts the equilibrium, resulting in the carbonate ion
formation as,
HCO3- + H++2NH4+ +2OH- CO32- +2NH4+ + 2H2O
The carbonate formed in reaction 6 was subsequently
reacted with Ca2+ ions deposited on bacterial cell wall,
leading to CaCO3 precipitation.
Cell-Ca2+ + CO32- Cell- CaCO3
MATERIALS AND METHODS
Materials and Chemical used- Soil samples used for
isolation of potent strain were collected from 12
different construction sites across the Gondia district,
India. All the required chemicals in this work were
purchased from Hi-media laboratories, India. The
research work was carried out in the P.G Department of
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Jokyani and Chouhan, 2018
DOI:10.21276/ijlssr.2018.4.5.10
Microbiology, Dhote Bandhu Science College, Gondia,
India from August 2017 to March 2018.
Preparation and artificial cracking of concrete blocks-
The concrete blocks of M20 grade were first prepared
with the mould size of 100 mm x 60 mm and left for 28
days curing process. Further, the blocks were subjected
to indigenous cracking by plastic shrinkage method
(plastic shrinkage cracks usually ranges between 1-2 mm
depth) [8].
Determining Size of cracks- Size of the cracks in concrete
block was measured using scale [9].
Isolation of new ureolytic bacteria- Soil samples
pasteurized at 800C for 15 minutes then subjected to
tenfold dilution. The 0.2 ml of pasteurized diluted sample
1 was then plated on sterile urea agar plates with 5% urea
concentration. Plates were incubated at 370C for 24 - 48
hrs and observed the isolated colonies. Pink color
colonies were selected and further screened based on
the capability to degrade urea in Christensen’s agar
2 medium and to tolerate high alkaline condition based on
pH optimization studies. The single selected potent
organism was in turn identified based on its
morphological and biochemical characteristics. Latter on
the identified isolate was pure cultured on urea agar
4 slants and preserved at low temperature (40C) [10].
5 Study on effect of environmental and nutritional
condition on urease production a capability of BH3
strains- Bacillus subtilis strain with promising result was
selected and the effect of environmental condition like
temperature, and nutritional condition such as urea
concentration were characterized. For temperature
6
optimization, 1 ml of overnight growth culture was
inoculated in 3 tubes containing urease broth except the
control tube and incubated at 3 different temperature
ranges as 250C, 350C, and 450C then after 24 hours the
tubes were observed for change in color intensity and
7 biomass was determined nephlometrically. For urea
concentration, optimization purpose 1 ml of overnight
growth culture was inoculated in 3 tubes containing urea
in concentration range of 2%, 5%, and 10% except the
control tube and incubated at 350C. After 24 hours the
tubes were observed for change in color intensity and
biomass was determined nephlometrically. [11]
 Int. J. Life. Sci. Scienti. Res.
Growth Curve of Bacillus subtilis strains: The growth
curve of screened isolate was determined to set a
growth comparison point at optimum condition. Three
ml of overnight growth culture was used to inoculate in
30 ml of urea broth in 250 ml conical flask then the
culture was incubated for 27 hours at 350C, pH 8, and 5%
urea concentration. Inoculation time was considered as
zero time and the quantitative determination of growth
was carried out by spectrophotometer at 600 nm.
Simultaneously the viable cell count was determined as a
colony forming units/ml (CFUs) [12]. The desired isolate
was purified and mixed with silica gel in 1: 100 dilution
factors with the cell count 104, 105, 106 cells so as to
immobilize the bacterial cells and hence it can remain
embedded for the long time in the concrete matrix.
Along with it, calcium lactate is also added to it with
molar mass 218 g/mole that serve as food source for
bacterial cells. As it gets the growth favorable conditions
it will soon start to precipitate CaCO3 and the bacterial
cells will be in turn coated with it resulting in the
autogenous healing [13].
Water Permeability Test of Concrete- To determine the
water absorption ability of bacterial concrete, this test
was carried out in a comparative manner. For this, the
control and bacteria treated concrete blocks were kept
overnight saturated in the saline buffer and weighed.
Then both blocks were dried in the oven at 1000C for 24
hours, cooled and again weighed. The obtained values
were then put into the formula given below to
determine percent water absorption by both the
eISSN: 2455-1716
Jokyani and Chouhan, 2018
DOI:10.21276/ijlssr.2018.4.5.10
[14]
blocks .
WSaturation – WOven Dried
% water absorption = X 100
WOven Dried
Where,
WSaturation = Weight of block after saturation
WOven Dried= Weight of block after drying
RESULTS
As the durability of concrete is affected by the cracks
leading to corrosion of reinforcing bars, the general
method of repairing is time consuming and expensive, so
bio based calcite precipitation has been proposed as an
alternative and sustainable, environment friendly cracks
repair technique. The bio agent selected for this purpose
was based on its tolerance to high pH, and continuous
formation of dense CaCO3 in the liquid medium.
According to analysis and study of blocks by the visual
examination of concrete core, the estimated size of
cracks varied from 0.3 to 0.5 mm [9]. Total 12 urease
positive Bacillus subtilis strains were isolated from soil
sample. Out of these 12 isolates Bacillus subtilis BH3
strains were found to be most potent.
Optimization- The optimum temperature required for
growth of Bacillus subtilis was 350C. Table 1 and Fig. 1
shows the optimization of temperature by using 3
different temperature ranges.

Table 1: Measurement of bacterial growth at 3 different temperature ranges

Growth measure
Temperature range
S. No. Bacterial strains Ureolytic activity Interpretation
(0C) OD600
Color changes to The maximum
1. 25 1.51 ureolytic activity
mild pink
with effective
Color changes to
2. BH3 35 1.52 growth was
intense pink
observed at
Color changes to temperature range
3. 45 1.519
light pink of 350C
BH3= Bacillus subtilis

The specific urea concentration supporting the growth Table 2 and Fig. 2 show the effect of urea concentration
of selected ureolytic bacterial strains (BH3) was 5%. The on the bacterial growth and urea degrading ability of it.

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Jokyani and Chouhan, 2018
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Effect of Temperature on growth of BH3

strain

Absorbance at 600 nm
1.525
1.52
1.515
1.51
1.505
25 35 45
Temperature range (0C)

Fig. 1: Effect of temperature on growth of Bacillus subtilis strains

Table 2: Measurement of bacterial growth at 3 different urea concentrations

Bacterial Urea Growth measure

S. No. Ureolytic activity Interpretation
strains concentration OD600

1. 2% 0.54 Color changes to light pink The maximum

ureolytic activity
Color changes to Intense with effective
2. BH3 5% 1.75
pink growth was
observed at 5% urea
3. 10% 1.69 Color changes to mild pink
concentration

Effect of urea concentration on growth of BH3

Absorbance at 600 nm

strain
2

1.5

0.5

0
2% 5% 10%
Urea concentration

Fig. 2: Effect of urea concentration on growth rate of Bacillus subtilis strains

Spectroscopic growth curve- The growth curve of referred as log phase. Nephlometric reading showed that
Bacillus subtilis was elucidated using the partially the cultures reached the stationary phase after 10 hours.
optimized condition. Table 3 and Fig. 3 indicate the After 24 hours the bacterial growth was inhibited due to
growth curve of B. subtilis isolates, which is considered media component depletion and the release of
as the selected isolate of this research paper. secondary metabolite that may be toxic to viable cells.
The maximum OD was seen between 6 to 10 hours and
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Table 3: Nephlometric absorbance values to construct growth curve of screened isolate of B. subtilis

Absorbance at 600 nm
Time in hours Trial 1 Trial 2 Trial 3 Average
1 0.162 0.164 0.180 0.168

2 0.372 0.343 0.417 0.377

3 0.651 0.621 0.682 0.651

4 0.911 1.001 1.073 0.995

6 1.400 1.545 1.521 1.488

8 1.618 1.647 1.690 1.651

9 1.671 1.677 1.700 1.682

10 1.720 1.677 1.710 1.702

11 1.723 1.694 1.721 1.712

12 1.731 1.717 1.729 1.725

25 1.019 1.724 0.940 1.227

26 0.911 0.853 0.848 0.870

27 0.908 0.847 0.769 0.841

Growth curve of strain BH3

2
OD at 600 nm

1.8
1.6
1.4
1.2
1
0.8
0.6
0.4
0.2
0
1 2 3 4 6 8 9 10 11 12 25 26 27

Hours of incubation

Fig. 3: Growth curve of screened potent isolate of B. subtilis strains

Studies on concrete crack healing potential of BH3
strains- In this investigation, the 3 concrete blocks, Block
1, Block 2, and Block 3 were used for autogenous healing
by the selected strains of Bacillus subtilis. Block 1 with
crack size of 0.45 mm width was injected with 105 CFU/
ml along with silica gel and calcium lactate showed the
healing of cracks at 21st day of inoculation and the width
of crack was reduced to 0.18 mm.
Similarly the Block 2 with crack size of 0.52 mm when
injected with 106 CFU/ml showed the efficient healing on
19th day of inoculation, whereas the Block 3 with crack
size 0.48 mm width, when injected with 104 CFU/ml,
showed the healing on 23rd day of inoculation with the
reduction in crack size up to 0.25 mm.

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Int. J. Life. Sci. Scienti. Res. eISSN: 2455-1716
Jokyani and Chouhan, 2018
DOI:10.21276/ijlssr.2018.4.5.10
5
Table 4: Concrete crack healing potential of B. subtilis strains in block 1 (10 cells/ml)

S. No. Date Initial Characteristics Final characteristic (Width) Interpretation

1. 22nd January 0.45 mm

2. 23rd January 0.45mm

3. 24th January 0.44 mm
4. 25th January 0.43 mm
5. 26th January 0.41 mm
6. 27th January 0.4 mm
7. 28th January 0.4 mm
8. 29th January 0.38 mm
9. 30th January 0.37 mm
Block 1, inoculated
10. 31st January Grade: M20 0.37 mm
with the 105 cells/ml
11. 1st February Size: 100 X 60 mm 0.35mm of BH 3 strains,
Curing: 28 days showed the
12. 2nd February Crack developed by: 0.34 mm
effective healing
13. 3rd February plastic shrinkage type 0.31 mm within 21 days from
Width: 0.45 mm the day of
14. 4th February 0.28 mm
inoculation
15. 5th February 0.27 mm
16. 6th February 0.25 mm
17. 7th February 0.23 mm
18 8th February 0.2 mm
19. 9th February 0.19 mm
20. 10th February 0.19 mm
21. 11th February 0.18 mm

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Jokyani and Chouhan, 2018
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Fig. 4: Crack healing study in bacteria treated cracked concrete block (Block 1)

Table 5: Concrete crack healing potential of B. subtilis strains in block 2 (106 cells/ml)

Block characteristics Final characteristic

S. No. Date Interpretation
(Width)
1. 22nd January 0.52 mm
rd
2. 23 January 0.52 mm
th
3. 24 January 0.52 mm
th
4. 25 January 0.518mm
th Grade: M20 Block 2, inoculated with the
5. 26 January 0.516 mm
Size: 100 X 60 mm 106 cells/ml of BH 3 strains,
6. 27th January Curing: 28 days 0.514 mm showed the effective healing
th
7. 28 January Crack developed by: 0.51 mm within 19 days from the day of
th
8. 29 January plastic shrinkage type 0.48 mm inoculation
th Width: 0.52 mm
9. 30 January 0.47 mm

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10. 31st January 0.47 mm

st
11. 1 February 0.46 mm
nd
12. 2 February 0.44 mm
rd
13. 3 February 0.44 mm
th
14. 4 February 0.4 mm
th
15. 5 February 0.4 mm
th
16. 6 February 0.4 mm
th
17. 7 February 0.39 mm
th
18. 8 February 0.39 mm
th
19. 9 February 0.38 mm

Fig. 5: Crack healing study in bacteria treated cracked concrete block (Block 2)

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Int. J. Life. Sci. Scienti. Res. eISSN: 2455-1716
Jokyani and Chouhan, 2018
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4
Table 6: Concrete crack healing potential of B. subtilis strains in block 3 (10 cells/ml)

S. No. Date Block characteristics Final characteristic (Width) Interpretation

1. 22nd January 0.52 mm

2. 23rd January 0.52 mm
3. 24th January 0.52 mm
4. 25th January 0.518mm

5. 26th January 0.516 mm

6. 27th January 0.514 mm

7. 28th January 0.51 mm

8. 29th January 0.48 mm

9. 30th January 0.47 mm

10. 31st January Grade: M20 0.47 mm

Size: 100 X 60 mm Block 3, inoculated with the
11. 1st February 0.46 mm
Curing: 28 days 104 cells/ml of BH3 strains,
12. 2nd February Crack developed by: 0.44 mm showed the effective healing
plastic shrinkage within 23 days from the day
13. 3rd February 0.44 mm
type of inoculation
14. 4th February Width: 0.48 mm 0.4 mm

15. 5th February 0.4 mm

16. 6th February 0.4 mm

17. 7th February 0.39 mm
18. 8th February 0.39 mm
19. 9th February 0.38 mm
20. 10th February 0.26 mm
21. 11th February 0.26 mm
22. 12th February 0.25 mm
23. 13th February 0.25 mm

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Jokyani and Chouhan, 2018
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Fig. 6: Crack healing study in bacteria treated cracked concrete block (Block 3)

Water absorption test by using saline buffer- The saline
buffer is used in this research work so as to determine
the increased resistance of concrete block towards the
water penetration and this test was conducted at a
laboratory level. According to this test, the normal
concrete block had shown the higher water absorption
as compared to the concrete block treated with potent
ureolytic bacterial BH3 strains. Table 7 numerically
represents the bacterial influence on the water
permeability of blocks.

Table 7: Water saturation test results of bacteria healed cracked concrete

Blocks No. Weight (Saturation) Weight (Dried) Water absorption (%)

Block 1 192.73 183.70 4.91%

Block 2 188.05 174.80 7.58%

Block 3 152.30 140.10 8.70%
Control 131.1 119.4 9.79%

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Int. J. Life. Sci. Scienti. Res.
DISCUSSION
This study has revealed the BH3 strains of bacterial
species (Bacillus subtilis) isolated from commercial
construction sites, have the high ureolytic activity and
can tolerate the pH range up to 12, which becomes the
primary factor for bio-cementation. Salmabanu L and
Suthar G [2] stated this genus of endolithic bacteria can
resist the pH range up to 13 and persist for the long
duration with efficient ureolytic activity. The isolated and
screened potent isolate (BH3) was identified
phenotypically and biochemically same as done by Achal
et al. [11] and Cheng et al. [15] while the species level
characterization was achieved by 16s gene sequencing
technique. From the further optimization studies of BH3
strains, it was clear that it shows the effective urea
degrading activity at temperature 350C with 5% urea
concentration. Similar conclusion was given by Steubing
[12]
, who stated that change in temperature and
nutritionally sound components can also affect the
CaCO3 precipitation efficiency by ureolytic bacteria.
Growth curve of this potent strains were determined
nephlometrically that showed the log phase of these
strains during initial 6-10 hours and become stationary
after 10 hours further reduction in the growth rate after
24 hours. The cell count in the range 105 CFU/ml when
injected in block 1 with the crack width size of 0.45 mm
shows the significant reduction in crack size up to 0.18 in
21 days after inoculation which was more effective as
compared to other 2 concrete block injected with 106
and 104 CFU/ml cells. A similar finding was reported by
Bai and Varghese [16] during an experimental
investigation on the strength properties of fly ash based
bacterial concrete.
The bacteria treated concrete block shows the significant
reduction in its water penetration ability compared to
normal concrete block due to the deposition of CaCO3
matrix like structure around the bacterial cells thereby
blocking the pores and inhibiting the water percolation
through it. As with constant development in the field of
civil engineering and increasing CO2 emission,
considerable efforts has been devoted to this alternative
concept of bio healing of concrete.
CONCLUSIONS
Our paper describes that due to its self-healing abilities,
eco-friendly nature, resistance to water absorption
thereby enhancing the strength of building materials
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along with prevention of steel corrosion, the microbial
concrete technology had proved to be better than the
conventional technologies. It may also be termed as
“Smart Bio Material”. The overall development of
strength in bio-concrete is attributed to significant
reduction in water absorption capability by using potent
bacterial strains of Bacillus subtilis immobilized by means
of silica gel and supplemented with calcium lactate as
food source. The efficient healing was shown with 105
cells, in block 1 with the reduction in crack size from 0.43
mm to 0.29 mm width at 21st day of inoculation.
More exploratory works at large scale should be
undertaken to determine the efficacy of bio cementation
for consolidations of building. To make the process
economical, microbial additives can repaired by
industrial growth of cells by employing the products as
lactose mother liquor and corn step liquor as nutrient
source. Also, the durability of bacterial concrete should
be studied under the various weathering conditions.
ACKNOWLEDGMENTS
The author is grateful to P.G. Department of
Microbiology, Dhote Bandhu Science College, India for
their help and support in carrying out the present study.
Technical assistance and advice was provided by Dr. D. A.
Chouhan so I would like to thank him for his
encouragement.
CONTRIBUTION OF AUTHORS
Research concept- Prof. D. A. Chouhan
Research design- Prof. D. A. Chouhan
Supervision- Prof. D. A. Chouhan
Data collection- Diksha H. Jokyani
Data analysis and interpretation- Diksha H. Jokyani
Literature search- Diksha H. Jokyani
Writing article- Diksha H. Jokyani, Prof. D. A. Chouhan
Critical review- Prof. D. A. Chouhan
Article editing- Prof. D. A. Chouhan
Final approval- Prof. D. A. Chouhan
REFERENCES
[1] Jayaraj G, Tapsi MS. Self-Healing of Concrete Using
Polymer Technique. International Journal of Recent
Engineering and Development, 2017; 2(1): 21-30.
[2] Salmabanu L, Suthar G. A Review on Self-Healing
Concrete. Journal of Civil Engineering Research,
2015; 5(3): 53-58.
Int. J. Life. Sci. Scienti. Res.
[3] Dinesh S, Shanmugapriyan R, Namitha Sheen ST. A
Review on Bacteria-Based Self-Healing Concrete.
Imperial Journal of Interdisciplinary Research, 2017;
3(1): 2454-1362.
[4] Jonkers HM, Thijssen A, Copuroglu O, Schalangen.
Application of bacteria as self-healing agent for the
development of sustainable concrete. Proceedings of
the 1st International Conference on Bio. Geo. Civil
Engineering, 2008; 36(2): 230-235.
[5] Santhosh K, Ramachandran SK, Ramakrishnan V,
Bang S. Remediation of concrete using
microorganisms. American Concrete Institute
Materials Journal, 2001; 98: 3-9.
[6] Bang SS, Galinat JK, Ramakrishnan V. Calcite
precipitation induced by polyurethane-immobilized
Bacillus pasteurii. Enzyme and Microbial Technology,
2001; 287(4): 404-409.
[7] Reinhardt HW, Joo M. Permeability and self-healing
of cracked concrete as a function of temperature and
crack width. Cement and Concrete Research, 2003;
33: 981-985.
[8] Dick J, De. Windt W, De Graef B, Saveyn H, Vander
Meeren P, De Belie N, Verstraete W. Bio-deposition
of a calcium carbonate layer on degraded limestone
by Bacillus species, Biodegradation, 2006; 17(4):
357-367.
[9] Shetty MS. Concrete Technology theory and practice.
Revised ed., India; Schand publication: 2006; p. 361-
362.
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Jokyani and Chouhan, 2018
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[10]Sahmaran M, Keskin SB, Ozerkan G, Yaman IO. Self
healing of mechanically-loaded self consolidating
concretes with high volumes of fly ash. Cement and
Concrete Composites, 2008; 30(10): 872-879.
[11]Achal V, Mukherjee A, Reddy S. Characterization of
two urease-producing and calcifying Bacillus spp.
isolated from cement. J. Microbiol. Biotechnol, 2010;
20(11): 1571–1576.
[12]Steubing P. Isolation of an Unknown Bacterium from
Soil. Proceedings of the 14th Workshop/Conference
of the Association for Biology Laboratory Education
(ABLE), 1993; p. 240.
[13]Kantha DA, Sathyanarayanan KS, Darshan BS, Raja
RB. Studies on the characterization of Biosealant
properties of Bacillus sphaericus. International
Journal of Engineering Science and Technology,
2010; 2(3): 270-277.
[14]Irwan JM, Faisal Alshalif A, Othman N, Anneza LH.
Effect of Ureolytic Bacteria on Compressive strength
and Water Permeability on Bio-concrete.
International Conference on Civil, Architectural,
Structural and Construction Engineering, 2015.
[15]Cheng L, Cord-Ruwisch R. In situ soil cementation
with ureolytic bacteria by surface percolation.
Ecological Engineering, 2012; 42: 64-72.
[16]Bai CP, Varghese S. An experimental investigation on
the strength properties of fly ash based bacterial
concrete. International Journal of Innovative
Research in Advanced Engineering, 2014; 3(8):
64-69.