Microbial Concrete- A Sustainable Solution for Concrete Crack Repair and Rehabilitation

Dr M V Seshagiri Rao, FIE, Professor in Civil Engineering, JNTUH College of Engineering, Hyderbad-85
V Srinivasa Reddy, MIE, Associate Professor in Civil Engineering, GRIET, Hyderbad-90

Abstract:
Introduction
Modern man-made engineering materials demonstrate excellent mechanical properties, but the
lack of the ability of self healing, i.e. the ability to remove or neutralize microcracks without
(much) intentional human interaction, which is typical for most materials as encountered in
nature. In contrast, natural materials, such as skin, wood, bones and skeletons, grass, etc, often
have the special ability that they can, more or less autonomously, heal cracks and other forms of
accidental damage.
Routes and mechanisms towards self healing behavior in engineering materials
In this paper the focus is on the physical or chemical mechanism used to obtain autonomous or
stimulated healing. In the routes described above all healing actions were due to physico
chemical reactions, not involving intermediate agents to control or accelerate the healing
reaction. However, in one route the use of calcium carbonate-producing bacteria, as agent to
control the healing action in
concrete, has been demonstrated successfully. Efficient sealing
of surface cracks by mineral precipitation was observed when
bacteria-based solutions where externally applied by spraying
onto damaged surfaces or by direct injection into cracks.
These treatments resulted in regained material strength and reduction
of surface permeability [48, 49]. However, in the latter
studies bacteria or their derived enzymes were not applied as
a truly self healing system but rather as an alternative, more
environmental friendly, repair system. To create autogenous
self healing behaviour, at the Technical University Delft the
option of using viable bacteria as a matrix-embedded healing
agent was explored [50, 51]. Major challenge in the latter
approach was to identify bacteria and their needed metabolic
components which are not only sustainable, but which also
do not negatively influence other concrete characteristics. Moreover, this biological system must
also feature a longterm
self healing functionality, preferably in agreement with
the constructions service life. In the latter studies a specialized
group of alkali resistant spore-forming bacteria affiliated
to the genus Bacillus was identified as an ideal self healing
agent as the spores of these bacteria appeared particularly resistant
to concrete incorporation and, moreover, feature life
spans of over 100 years [52]. Furthermore, once incorporated
in concrete, these bacteria appeared able to convert various

natural organic substances to copious amounts of large, over

100-m sized, calcium carbonate-based crystalline precipitates
Such a bacteria-based self-healing mechanism thus appears
to be a promising alternative to non-sustainable cementbased
healing systems particularly because the formation of
large crystalline precipitates potentially enables sealing of
larger than 100 m-wide cracks. The formation of large precipitates
can be explained by the high local bacterial CO2
production rates. Due to conversion of CO2 into carbonate
ions under alkaline conditions and subsequent reaction with
free calcium ions leached from the concrete matrix calcium
carbonate-based precipitates are formed. Furthermore, locally
produced CO2 directly reacts with matrix portlandite (calcium
hydroxide) crystals which contributes to calcium carbonatebased
mineral formation. The intrinsic CO2-producing capacity
with the concomitant production of large-sized precipitates
in the bacteria-based self healing concrete may result in a superior
self healing rate and capacity compared to traditional or
engineered non-sustainable self healing cementitious systems.
The beneficial effect of bacterial closure of the deformation
induced cracks has been demonstrated in water permeability
tests. Even for quite substantial cracking the bacterial self
healing concrete showed no residual permeability after allowing
two weeks for crack healing. Such a restoration of
water impermeability may be as important as the restoration
of mechanical properties, in particular for underground constructions
(tunnels, basements and underground garages) in
sloppy water carrying soils. While several technical issues
yet need to be solved, the route of bacteria supported self
healing concrete seems very promising and is now entering
a pre-commericalisation stage.
To turn these experimental approaches
under laboratory conditions into commercially available
materials and products, it is necessary to initiate research into
the quantification of the healing behaviour as a function of
the damage characteristics and the healing mechanisms. Such
an extension is now becoming overdue.
Effect of ureolytic bacteria on concrete properties
Recently, microbial mineral precipitation resulting from metabolic activities of some specific
microorganisms in concrete to improve the overall behavior of concrete has become an important
area
of research. It has been hypothesized that almost all bacteria are capable of CaCO3 production
because

precipitation occurs as a byproduct of common metabolic processes such as photosynthesis,

sulfate
reduction, and urea hydrolysis.
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).
Introduction
The applicability of specifically mineral producing bacteria for sand consolidation and limestone
monument repair [48] and filling of pores and cracks in
concrete have been recently investigated [912]. In all these studies
so far, bacteria or derived ureolytic enzymes 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 environmental friendly repair material is
needed.
Microbial carbonate precipitation (biodeposition) 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.
In the laboratory, bacteria are usually grown using solid or
liquid media. Solid growth media such as agar plates are used to
isolate pure cultures of a bacterial strain. However, liquid growth
media are used when measurement of growth or large volumes
of cells are required. Growth in stirred liquid media occurs as an
even cell suspension, making the cultures easy to divide and transfer,
although isolating single bacteria from liquid media is difficult.
The use of selective media (media with specific nutrients added or
deficient or with antibiotics added) can help identify specific
organisms.
Most laboratory techniques for growing bacteria use high levels
of nutrients to produce large amounts of cells cheaply and quickly.

However, in natural environments nutrients are limited, meaning

that bacteria cannot continue to reproduce indefinitely. This nutrient
limitation has led the evolution of different growth strategies.
carbonate biomineralization
Biomineralization is 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
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.
Recently, microbial mineral precipitation resulting from metabolic
activities of some specific microorganisms in concrete to
improve the overall behavior of concrete has become an important
area of research. It has been hypothesized that almost all bacteria
are capable of CaCO3 production because precipitation occurs as a
byproduct of common metabolic processes such as photosynthesis,
sulfate reduction, and urea hydrolysis [15].
When this hydrolysis occurs in a calcium-rich
environment, calcite (calcium carbonate) precipitates from solution
forming a solid-crystalline material. 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 biocement).
Members of the genus Bacillus are Gram-positive,
rod-shaped, endosporeforming bacteria commonly found in soil
[26]. B. pasteurii, a member of this genus, converts urea to ammonium
carbonate more actively than any other known bacterium.
Therefore, B. pasteurii and other members of the Bacillus genus
are pincorporated into studies to determine their influence on calcium
carbonate precipitation in various environments.
Microbiologically induced (also called bacteriogenic) calcium
carbonate precipitation is comprised of a series of complex biochemical
reactions
The ammonia increases
the pH in surroundings, which in turn induces precipitation of
CaCO3, mainly as a form of calcite.

In aqueous environments, the overall chemical equilibrium

reaction of calcite precipitation can be described as [27]:
The solubility of CaCO3 is a function of pH and affected by ionic
strength in the aqueous medium.
Ca2+ is not likely utilized by microbial metabolic processes;
rather it accumulates outside the cell. In medium, it is possible that
individual microorganisms produce ammonia as a result of enzymatic
urea hydrolysis to create an 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. Possible biochemical reactions in
ureaCaCl2 medium to precipitate CaCO3 at the cell surface

Bacteria-based self-healing concrete

Crack formation in concrete is common, but a typical phenomenon
related to durability. Percolation of cracks may lead to leakage problems or ingress
of deleterious materials, causing deterioration of the concrete matrix or corrosion of
embedded steel reinforcement. Durability can be enhanced by preventing further
ingress of water and other substances. In recent years a bacteria-based selfhealing
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
an organic mineral precursor compound. Whenever cracks occur and water is
present the bacteria become active and convert the incorporated organic compounds
into calcium carbonate, which precipitates and is able to seal and block
cracks. This paper aims to review the development of bacteria-based self-healing
concrete, introducing the proposed healing system. Different stages in the development
are discussed, and some recommendations for further research are given.
Self-healing is characterized by regaining performance after a defect occurs.
Damage targeted in bacteria-based self-healing concrete particularly relates to
increased durability and leakage prevention and extending service life of concrete
structures. Jonkers (2007) introduced a two-component healing agent to be added
to the concrete mixture, consisting of bacteria and a mineral precursor compound.
Upon cracking the system is activated by ingress water. Bacteria convert the mineral

precursor compound into the mineral calcium carbonate, better known as

limestone. Precipitation of the limestone on the crack surface enables sealing and
plugging of the cracks, making the matrix less accessible to water and other deleterious
materials. New studies will focus on further development
of the system in order to make practical application of the material feasible.
Target for selfhealing
concrete is to reduce matrix permeability by sealing or blocking cracks.
Healing agent is incorporated in the concrete matrix and acts without human intervention. To
make the material technically and economically competitive, healing
agent should be cheap in relation to the low price of concrete, remain potentially
active for long periods of time and be concrete compatible to not negatively affect
material characteristics.
Microbial healing
Microbial Concrete Sustainable Solution to enhance concrete durability
Microbial concrete is a new type of sustainable, eco-friendly material that enhances
concrete performance by the microbial deposition of calcium carbonate within concrete
pores through molecular reactions reducing fissures and cracks. The use of these reactions
to improve concrete performance at the microscopic and macroscopic level is investigated
in this paper in terms of increase in strength and durability, environmental impacts, and
economical aspects. Experimental results have shown a 27% increase in compressive
strength at 28 days, a 13% reduction in volume of pores, and a 12% decrease in water
absorption in concrete specimens. These outcomes were attributed to the activity of the
bacteria, as observed by scanning electron microscopy (SEM). The cost analysis showed an
increase in cost relative to conventional concrete. Finally, the environmental analysis
showed a 26.8% reduction in carbon dioxide produced per cubic meter of mix concrete
projecting to a 28.6% decrease of carbon dioxide emissions by the cement industry on a
global scale.
INTRODUCTION
Concrete is one of the most widely used construction materials in the world, and is actually the
second most consumable product after water
Microbial concrete is produced by incorporating a certain species of calcium depositing bacteria,
Sporosarcina pasteurii, formerly known as Bacillus pasteurii , into concrete. This bacterial
species has also been investigated as a remedy for the liquefaction of soils under buildings, to
replace potentially harmful epoxy chemicals [4]. However, the advantages of using this
bacterium in concrete have only been recently discovered; and it is expected to have a positive
impact on concrete properties, particularly permeability, strength, cost, and the effect on
environment.
Sporosarcina pasteurii is used to deposit calcite, a sealing agent, in the pores in concrete by the
process of microbiologically induced calcium carbonate precipitation,

MICCP. The structure, distribution, and connectivity of these pores have a great influence on
concrete strength and durability.
Inside the bacterial cell, urease hydrolyses urea into ammonia and carbonate. The latter directly
decomposes, producing ammonia and carbonic acid. The resulting ammonia and carbonic acid
equilibrate in water, inducing an increase in pH. According to Le Chatellier principle the
equilibrium will shift in such away as to reduce the effect of the induced change. In this case the
bicarbonate equilibrium is shifted to produce carbonate ions at the vicinity of the cell, and
eventually in the entire solution as shown in (2).

From equation 1, the increase in the concentration of carbonate ions, finally leads to the
deposition of calcium carbonate crystals.
Several experiments have been conducted to investigate the effect of adding the bacteria to
concrete. Achal et Al.(2010) mixed samples of mortar cement with a culture of bacteria. The
compressive strength of the samples mixed with bacteria was increased compared to the control
samples.
Ramachandran et Al. (2001) studied the remediation of cracks and fissures by MICCP in
concrete beams. Results showed better resistance to harsh environments in microbial concrete
beams compared to control beams.
In this paper, experimental work has been performed to test the qualities of microbial concrete,
including compressive strength, permeability, and absorption. In addition, samples of microbial
concrete and conventional concrete were observed under a scanning electron microscope.
Furthermore, the implementation of this technology was examined in terms of cost and
environmental effects, specifically its effect on the global carbon footprint of the cement
industry.

MATERIALS AND METHODOLOGY

A. Concrete mix
The Portland cement bags were purchased locally from Holcim Liban, and were in accordance to
ISO 9001:2008. The coarse aggregates used were mainly composed of crushed angular limestone
purchased from a local quarry. Natural river sand with a fineness modulus of 2.82 was used as
fine aggregates.
B. Bacterial suspension
Bacteria known as Sporosarcina Pasteurii ATCC 11859 were used in this research. It was
purchased from the American Type Culture Collection, USA through the Environmental
Engineering Research Center at the American University of Beirut. In order to preserve the
bacterial culture, a subculture was performed every 48 hours on Ammonia-Yeast Extract agar
plates and incubated with no agitation at 30C.

As for the bacterial solution, colonies of Sporosarcina pasteurii were inoculated in an Nutrient
Broth-Urea (NBU) solution containing Nutrient Broth (0.8%), Urea (2%), Sodium Chloride
(0.5%) and Calcium Chloride (2.775gr. per Lit). The bacteria-NBU solution was agitated for 24
hours at 37C.
Several experiments in the structural laboratory were conducted showing that the highest 28 day
compressive strength increase is obtained with a concentration of 105 cells/ml. The concentration
of bacteria was validated using miles and misra- serial dilution.
C. Concrete Mixing and Curing
Two types of Concrete mixes were prepared for this research. The mixes had similar proportions
of sand, coarse aggregates, cement and water. However, the main difference was in the water
component: Tap Water was used for specimens without bacteria (Mix A), while NBU-Bacteria
solution was used for specimens with bacteria (Mix B).
D. Compressive Strength Test
Three specimens of each type were tested under compression at 7, 14 and 28 days after casting.
Research done by Achal (2011) and Chattopadhvav (2010) showed that no increase in strength
will occur using a NBU solution without bacteria hence this research did not include specimens
of that type.
E. Water Absorption and Volume of Voids Test
To determine the total water absorption and measure the volume of voids in hardened concrete
cylinders, a series of tests were conducted according to the ASTM C642-13.The following steps
were performed: .
1. 28 days after casting, the specimens were oven dried at around 110C for 24 hours, and the
oven dried sample mass was measured.
2. The specimens were immersed in water at 21 oC for 52 hours, until two successive values of
the surface-dried specimens mass showed an increase of less than 0.5% of the larger value. The
final surface dry mass after immersion was obtained.
3. The specimens were boiled for 5 hours, cooled for 15 hours, and the surface-dried specimen
was weighed.
4. The apparent mass of the specimen in water after boiling was found.
F. Scanning electron microscope (SEM)
To prove presence of bacteria in the microbial concrete, the samples were observed under a
scanning electron microscope (SEM) and compared to concrete samples without bacteria.
Broken pieces of 28 day old concrete from the compression test that had been dried at room
temperature for 48 hours were crushed to a maximum size of 1mm and attached to SEM stubs
using carbon tape. Two stubs of each type of concrete were prepared.
G. Cost analysis
The cost of using microbial concrete compared to conventional concrete is critical in determining
the economic feasibility of the technology. To compare cost, the compressive strength of
concrete samples was fixed at 38MPa. The microbial concrete mix (B38) gave a compressive
strength of 37.85 MPa when tested at 28 days. The concrete mix design and corresponding prices
are given in table 1 and those of the NBU-bacterial solution are given in table 2. A conventional

concrete mix (mix A38) was designed for a compressive strength of 38 MPa according to
Mamlouk and Zaniewski (2011). The mix design and prices are given in table 3.

H. Environmental analysis

One of the major advantages of microbial concrete is the possible reduction of the carbon
footprint of concrete production. The production of cement, particularly clinker production, is the
major contributor to carbon dioxide emissions [10]. The carbon dioxide producing reaction in
cement manufacturing is given in (3)
Portland cement consists of 95% clinker of which 64% to 67% is Calcium Carbonate CaCO3
[10]; According to equation (3) this results in approximately 0.264 Kg of carbon dioxide
emissions per kilogram of cement produced.
Since microbial concrete yields the same compressive strength with decreased cement content,
the amount of carbon dioxide emissions during the production of cement for 38MPa concrete
was obtained for Mix A38 and Mix B38 and compared. Consequently, the ratio of carbon dioxide
reduction was used to obtain the projected global decrease in emissions.
I. Calcium carbonate quantification
A chemical experiment is being performed in the Environmental Engineering Research Center in
order to track the deposition of Calcium Carbonate by the Sporosarcina Pasteurii in mortar
cement.
Calcium carbonate CaCO3 reacts with hydrochloric acid (HCl) to produce Carbon Dioxide gas
through the following equation (4):
HCl (aq) + CaCO3 (s) CaCl2(aq) + H2O(l) + CO2 (g) (4)
Two types of cement specimens were casted: For the first type, 1 Kg of cement was mixed with
360 ml of water and for the second type; 1 Kg of cement was mixed with 360 ml of NBUbacteria solution.
1 gram of each oven-dried sample was crushed to powder and tested at the 1st and 2nd days.
Tests will continue on the 3rd, 4th,5th,7th , 14th , and 28th days respectively.
The cementeous powder was led to a complete reaction with 200ml of a 1.2M HCl solution with
agitation. The volume of gas produced was collected under water in an inverted measuring
cylinder. The experimental set up is shown in Fig. 3. When bubbled into lime water, the gas

produced turned it from colorless to milky white, hence confirming the nature of the gas as

carbon dioxide.
II. RESULTS AND DISCUSSION
A. Compressive strength test
The following bar chart (Fig. 4) shows the compressive strength results at 7, 14 and 28 days for
concrete cylinders of types A and B.

At 7 days-The average compressive strength of both types is comparable, marked by a 0.92%

increase. Provided that the cylinders were somehow wet and the concrete did not completely
harden, the compressive strength is mainly associated to the weakness of humid concrete during
the early stages. The following results confirm that during the beginning of the curing period, the
precipitation of calcite is slow and Sporosarcina pasteurii is being gradually activated.
Furthermore, the results confirm that the same mixing process and design was adopted for both
sets of specimens. Therefore, the comparative analysis can be accurately validated based on this
set of specimens.
At 14 days- The 14th day strength results show an increase of 12.48% in the specimens
containing NBU-bacterial cells. This improvement is mainly due to the strengthening of concrete
material and deposition of CaCO3 filling small sized pores. During bacterial growth, the calcium
precipitation process occurs continuously, clogging the internal pores with calcium precipitate.
The calcium carbonate precipitate prevents oxygen from reaching the bacterial cells leading to
their death. The cells are transformed into endospores that act as organic solid fibers.
The results are considered to be consistent since all obtained values are close within a range of
1.1 Mpa, and are within 60%-70% of maximum strength.
At 28 days- At this stage, the concrete has completely hardened, small and big size pores are
filled with bacteria. CaCO3 crystals were formed and developed on the external bacterial surface
within the pore structure. The 27% strength increase is mainly due to the calcium deposition
within the pores of the sample. The values obtained vary within a range of 1.35 Mpa, hence the
results are considered properly accurate.
Fig. 5 summarizes the compression test results showing an increase in compressive strength
compared to control samples.
B. Water absorption and volume of voids test
Water absorption experiment was conducted on three cylinders of each type (A and B). After
being dried at 110 oC, the cylinders went through three 52-hour cycles of immersion and
weighing.
Fig. 6 shows the mass of water absorbed in each cylinder after the last cycle.
It can be clearly seen that specimens of type B containing bacteria, absorbed 12.30% less water
than the ones of type A. The reduction in mass of water absorbed is consistent throughout the
samples and varies between 13g and 20g.

The volume of permeable pores was also calculated after boiling 3 cylinders of each type. The
results are presented in Fig. 7 showing an average of 13.1% decrease in the volume of permeable
pores.

The experimental results are associated to the decrease in the number of pores within the
concrete internal structure due to the calcium carbonate precipitation.
C. Scanning electron microscope (SEM)
Rod shaped structures, which were perceived to be Sporosarcina pasteurii, were observed in
both specimens with bacteria (type B). On the other hand, no such structures were found in the
samples without bacteria (type A). Fig. 8 shows a microbial concrete micrograph and Fig. 9

shows conventional concrete micrograph. Since the only difference between the micrographs is
the presence of the rod like structures, it is most likely that the activity of these structures has
caused the 27% increase in compressive strength that was observed at 28 days.
D. Cost Analysis
The cost analysis showed an increase in cost of 4.9 times between microbial concrete and
conventional concrete (mix B38 and A38 respectively). The major contributor to the cost of the
B38 mix is the nutrient broth, amounting to over 70% of the cost per cubic meter of concrete.
Therefore, further research needs to be devoted to decrease the amount of the nutrient broth used
or to find a cheaper alternative source for bacteria nutrition in order to make the technology
financially feasible. Nevertheless, it is believed that microbial concrete will yield cost reductions
on the long run through decreased need for rehabilitation and maintenance. Furthermore, the
high cost of the technology is outweighed by its positive environmental impact, which is
discussed in the following section.
E. Environmental Analysis
According to the calculations explained in the methodology, Mix A38 yields 125.4 kg of carbon
dioxide during cement production while Mix B38 yields 89.5 kg of carbon dioxide per cubic
meter of concrete, amounting to a 26.8 % decrease in carbon dioxide emissions per cubic meter
of concrete.
Using the percent reduction obtained above when using the NBU-bacterial solutions, the
reduction in carbon dioxide emissions was projected to the global scale yielding the data given in
Fig. 10.
There is a 28.6% reduction of cement related carbon dioxide emissions on a global scale,
specifically from 219.7 to 156.8 megatons of carbon dioxide by the cement industry.
F. Calcium carbonate quantification
The results for the experiments are presented in table 4. Over all, there is an increase in the
amount of gas produced in the samples containing bacteria showing that there is more Calcium
Carbonate in these samples. Since the samples have the same proportions of cement and water
the results are attributed to the presence of Sporosarcina pasteurii. Furthermore, to make sure
that the nutrient broth solution does not produce carbon dioxide when reacted with HCl, the same
test was done using NBU-bacterial solution and acid only; no gas was produced.

III. WORK IN PROGRESS

In addition to the experiments and results described earlier, this section outlines further
experiments that will be conducted, namely freeze and thaw, the effect of the bacterial solution
on steel, and splitting tensile strength test.

A freeze and thaw experiment will be conducted according to ASTM C67. The experiment
requires 15 freeze and thaw cycles for 15 days. In each cycle, the concrete will be placed in a
plate that contains 0.5 inch of water and then subjected to 16 hours of freezing at -4 1o C,
followed by 8 hours of thawing at room temperature. After the 15 cycles have been completed,
the compressive strength of the concrete will be determined.
Furthermore, the effect of microbial concrete on steel will be examined to ensure the feasibility
of the technology in reinforced concrete. Steel will be placed in the medium containing bacteria
and cement powder for a certain time period. The steel will be removed at regular time intervals
and any resulting rust will be analyzed. The results obtained will be compared to two control
experiments, in the first, the steel is placed in water and cement powder and in the second, the
steel is placed in cement and NBU-solution.
Finally, a tensile split test will be performed on concrete cylinders in order to assess the effect of
microbial calcium carbonate precipitation on tensile capacity
IV. CONCLUSION
The purpose of this study is to examine the properties of microbial concrete in addition to its cost
and its effect on the environment compared to conventional concrete. Experimental results have
shown increased compressive strength and decreased absorption and porosity which were
attributed to the precipitation of Calcite by the Sporosarcina pasteurii that was observed under
the scanning electron microscope. In addition, reacting the microbial mortar with hydrochloric
acid yielded more carbon dioxide than with control samples, indicating the presence of more
calcium carbonate. Furthermore, analysis has shown an increase in the cost of production and a
significant decrease in carbon footprint compared to conventional concrete. Microbial concrete is
thought to be a promising innovation. Nevertheless, research has to be devoted to finding
methods to decrease the cost of production and to implement microbial concrete on an industrial
scale to ensure the success of the technology.
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Executive
summary
[Online].
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[8] B. Chattopadhyay, "Bioremediase a unique protein from a novel bacterium BKH1, ushering a
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Bio-based Crack repair and Performance enhancements of Microbial Concrete
Application of bacteria-based repair system to damaged concrete structures
The goal of this study is to present the development of bacteria-based repair system which
features improved durability and sustainability characteristics compared to currently
commercially available
systems.
Concrete is strong and relatively cheap, but it is also
subjected to a number of degradation processes
which hamper the structure to reach its required service
life. To anticipate durability problems during
the lifetime of a structure, costly measures of
maintenance and repair have to be undertaken.
Currently available concrete curing and repair
system aiming to decrease porosity and repair of
cracks in aged concrete structures are largely based
on environmental unfriendly materials systems.
Moreover, periodic maintenance operations for concrete
structures are generally focused on repairing
concrete damages while not considering the relevant
durability issues of the repair system itself (Robery,
2011; Tilly and Jacobs, 2007).
Working principle of bacteria-based repair system
The bacteria used are alkaliphilic
species from the genus Bacillus which
grow in alkaline environment such as in concrete.
Different pathways appear to be involved in
calcium carbonate precipitation.
The first pathway involves the sulphur cycle, in
particular sulphate reduction, which is carried out
by Advanced Topics in Bio mineralization sulphate
reducing bacteria under anoxic conditions.
A second pathway involves the nitrogen cycle,
and more specifically,

(1) The oxidative deamination of amino acids in

aerobiosis,
(2) The reduction of nitrate in anaerobiosis or
microaerophily and
(3) The degradation of urea or uric acid in
aerobiosis (by ureolytic bacteria).
Finding Right Bacteria:
The starting point of the research is to find bacteria capable of
surviving in an extreme alkaline environment. Cement and
water have a pH value of up to 13 when mixed together,
usually a hostile environment for life most organisms die in an
environment with a pH value of 10 or above [13]. The search
concentrated on microbes that thrive in alkaline environments
which can be found in natural environments, Samples of
endolithic bacteria (bacteria that can live inside stones) will be
collected along with bacteria found in sediments in the lakes.
Strains of the bacteria genus Bacillus will be found to thrive in
this high-alkaline environment. Different types of bacteria
which can survive in such a high Ph environment is mentioned
in Table 1. It is found that the only group of bacteria that will
be able to survive is the ones that produced spores comparable
to plant seeds. Such spores have extremely thick cell walls that
enable them to remain intact for up to 200 years while waiting
for a better environment to germinate. They would become
activated when the concrete starts to crack, food is available,
and water seeps into the structure [19]. This process lowers the
pH of the highly alkaline concrete to values in the range (pH
10 to 11.5) where the bacterial spores become activated.
Preparation of Microbial Cement Paste and
Mortar Cubes:
5cm3 moulds were used. Cement paste and mortar (cement:
sand, 1:3) cubes were casted by mixing grown bacterial
cultures of concentration 105, 106, 107, and 108 cells/ml of
water, at a W/C ratio of 0.4. Conventional cement paste
specimens with regular water are also casted parallel. The
cubes were cured under tap water at room temperature and
tested at 1, 7, and 28 days.
Preparation of Concrete Samples with Bacteria:
Bacterial concrete casted by using ordinary Portland cement
mixed with bacterial concentration 106 cells/ml of water.
Conventional concrete samples were also casted in parallel.
The specimens were cured under tap water at room
temperature and tested at 7, and 28 days.

Introduction1
Self-healing concrete is defined as the ability of
concrete to repair its small cracks autonomously [1]. Idea
of self-healing concrete is inspired from natural
phenomenon at organisms such as trees or animals.
Damaged skin of trees and animals can be repaired
autonomously [2]. Remediating cracks in concrete
structure is necessary because cracks not only influence
the service durability, but also harmful for the structure
safety [3].
Recently developing of self-healing concrete
technology has been becomes an important objective of
researches in biotechnology and civil engineering area
Several processes have been suggested for selfhealing
concrete design. Self-healing processes are
classified into natural and man-made processes. Among
of proposed natural processes, formation of calcium
carbonate and calcium hydroxide are the most important
reason to heal concrete naturally [11-18]. Although,
Corresponding author: Muhd Zaimi Abd Majid,
Construction Research Alliance, Faculty of Civil
engineering, Universiti Teknologi Malaysia, UTM
Skudai, 81310 Johor Bahru, Malaysia, e-mail:
mzaimi@utm.my.
concrete has naturally ability of healing itself, only small
cracks can be naturally healed [19].
Chemical and biological processes (as man-made
processes) are useable to design self-healing concrete.
Many articles have been published about chemical and
biological self-healing concrete development. However,
number of articles on biological methods to design selfhealing
concrete is not considerable. Gollapudi et al.
introduced biological self-healing concrete as an
environmentally friendly process at the mid-1990s [20].
Especial strains of bacteria that are able to precipitate
especial chemicals such as polymorphic iron-aluminumsilicate
((Fe5AI3)(SiAl)Ol0(OH)5) and calcium carbonate
(CaCO3) are used to design a biological self-healing
concrete. Precipitation of these especial chemicals on the
concrete using microorganisms can reduce permeability
towards gas and capillary water uptake.
Although using of biological methods is an
environmental friendly, pollution free and natural way to
design self-healing concretes, these methods have some

disadvantages such as many prerequisites to be met,

measures should be taken to protect bacteria in concrete,
and mechanical properties recovery and effectiveness
under multiple damage events could be concerns [12].
Some researchers have been carried out by adding
microorganisms directly into fresh concrete. It is cheapest
way to conduct a research on biological self-healing
concrete [33]. pH of fresh concrete is between 10 to 13.
The temperature of fresh concrete can be also near 70
degree centigrade. After drying of concrete, there is not
enough water. Therefore, suitable bacteria have to have a
high resistance against high pH, temperature, and serious
limitation of water. Usually mesophilic microorganisms
cannot have a normal growing in this condition.
Dislike of bacteria, spore of bacteria is very
resistance against inappropriate condition and some
bacterial spores can live more than 60 years. Then in
some studies instead of direct using of microorganisms in
fresh concrete, spores were used. To avoid
microorganisms from inappropriate condition,
encapsulated microorganisms can be used. Encapsulation
of microorganisms is an expensive and complex way.
Using of vascular or microvascular networks to distribute
of a liquid contain microorganisms throughout of
concrete are other ideas to avoid microorganisms from
inappropriate condition. However, these methods are
extremely complex and they do not have constructability
using present technology.
The use of immobilized microorganisms onto silica gel or
activated carbon is a suitable way from aspect of cost.
However, effect of using these materials on the
strengthening of concrete is not completely clear.

Fig1.Shows direct stereomicroscopic observation of cracks

from control and bacteria-based specimens before and after
100 days of immersion in tap water. Width of completely
healed cracks was significantly larger in bacteria-based
specimens (0.46 mm) compared to control specimens
(0.18 mm).
ADVANTAGES AND DISADVASNTAGES
The Self Healing Concrete has comparatively very less
permeability, more durability and strain bearing capacity than
the conventional concrete.
A potential drawback of this reaction mechanism is
that for each carbonate ion two Ammonium ions are
simultaneously produced which may result in excessive
environmental nitrogen loading.
CONCLUSIONS
While most healing agents are chemically based,
more recently the possible application of bacteria
as self-healing agent has also been considered. In
a number of published studies the potential of
calcite precipitating bacteria for concrete or
limestone surface remediation or durability
improvement has been investigated.
Metabolically active bacteria consume oxygen;
the healing agent may act as an oxygen diffusion
barrier protecting the steel reinforcement against
corrosion. So far, bacteria have never been used to
remove oxygen from the concrete matrix to inhibit
reinforcement corrosion and further studies are
needed to quantify this potentially additional

beneficial process.
While, in this study many features of Self Healing
Concrete have been quantified but the life of
bacteria, cost of construction and efficiency still
needs a separate study.
Problem Statement
It was an eminent fact that concrete structures are extremely susceptible to microcracking which
allows water, gases and other potential harmful liquids enter and degrade the concrete, reducing
the performance of the structure in terms of strength and durability aspects. To defeat this
disadvantage it requires expensive continuous maintenance in the form of micro crack repairs.
When these microcracks propagate further deep, not only the concrete itself will be damaged, but
also leads to corrosion in the steel reinforced concrete structures. Microcracks are therefore the
major cause in reducing the durability of concrete structures. There are many crack repair
techniques available to surmount this problem but each technique has its own advantages and
disadvantages. Methods currently used for crack remediation often use synthetic polymers which
are expensive, incompatible, doubtful long-term performance, needs skilled human assistance
and aesthetically unpleasant (especially in repairing historic monuments) most importantly these
methods are not environment friendly and are very expensive. The idea of selfcrack healing
mechanism in concrete is developed to circumvent the above stated disadvantages. One such
alternative crack-repair mechanism is application of biominerilization of bacteria to seal and heal
cracks in concrete. Synthetic polymers such as epoxy treatment etc, at present being used for
repair of concrete, are harmful to the environment; therefore the use of a biological repair
technique in concrete was focused upon. The principle of self crack healing mechanism is that
certain kinds of healing agents will be released from the concrete when cracks occur. Every
repair method follows the procedure of detection, monitoring and repair of cracks but in the self
crack healing method the procedure of detection, monitoring and repair is autogeneous
throughout the structures life-cycle thus reducing the repair maintenance significantly. Repairs
can be particularly time consuming and expensive because it is often very difficult to gain access
to the structure to make repairs, especially if they are underground or at a great height. Currently
available concrete repair systems are largely based on environmental unfriendly material
systems. So research is focused on the biotechnology based crack remediation in concrete to
study the crack healing process of concrete with less or no human intervention and also examine
the effect of biogenic calcite precipitation on the mechanical and durability aspects of concrete
structures.
2.2.2 Research Significance
In the recent past, investigations attempted to study about the application of biomineralization in
civil engineering. As a part of those studies, researchers around the world started working on the
use of specific bacteria in cementitious materials to self-heal and seal cracks without human
intervention. Available literature has not reported any such suitable self-healing system which
has features such as long-term compatibility, eco-friendliness, good bonding with surrounding
cement matrix, less human intervention, inexpensive and organic in nature. Though it is reported
that the use of specific alkaliphilic mineral forming bacteria enhances the properties of cement
mortar but there exists little understanding of the effect of bacteria on the mechanical and

durability properties of concrete. In the present research work, studies related to characterization
of mineral precipitation, permeation properties, resistance to aggressive environment, resistance
to corrosion, behaviour at elevated temperature etc of bacteria incorporated concrete has been
reported.
Gaps in the Literature
Not much work has been reported on the use of microorganism Bacillus Subtilis JC3, isolated
and cultured at JNTUH Hyderabad, and its calcite mineral precipitation efficiency using the
nitrogen cycle as its microbial pathway in enhancing the mechanical and durability
characteristics of different grades of concrete for Indian conditions. An exhaustive comparative
study of mechanical and durability characteristics of ordinary grade (M20), standard grade
(M40) and high strength grade (M60 and M80) concrete with and without addition of bacteria
Bacillus Subtilis JC3 are made to understand the microstructure of bacteria incorporated concrete
at micro and macro level. Hence to address the gaps available in the research, investigations are
planned to study the effect of calcite mineral producing bacteria Bacillus subtilis JC3 on the
microstructure of various grades of concrete and its impact on its mechanical and durability
properties. Once the gaps available in the literature are identified, the main research objectives of
the present research work are outlined for obtaining detailed experimental data to address these
gaps, which will help to understand the bacteria incorporated concrete and its characteristics in
terms of strength and durability aspects under Indian conditions.
ASSESSMENT OF CRACK REPAIR EFFICIENCY
Studies on Crack Remediation and Strength Regain
Introduction
The aim of this test is to evaluate the efficiency of crack remediation technique using bacteria
Bacillus subtilis JC3 and determine the degree of strength regain in cracked concrete specimens.
Test methodology
To demonstrate the crack healing efficiency and strength regain, cracked cement mortar samples
were prepared in two different ways. The first method resulted in samples with standardized
cracks while the second method gave rise to more realistic cracked samples.
(a) Standardized cracks
Standardized cracks were made in twelve number of cement mortar cubes of size
70.6x70.6x70.6mm by making a cut to simulate a crack. The width of cut is kept at an average of
3mm width and a 20mm depth as shown in Fig 9.1. Cracks in the three cement mortar specimens
are filled up with Indian standard grade II sand (1mm to 0.5 mm) mixed with water and
specimens are cured in distilled water after air dried for one hr. The cracks in another set of three
specimens are closed with Indian standard grade II sand mixed with 10 5 cell concentration of
bacteria Bacillus subtilis JC3 and after air dried for one hr, cubes are cured in bacteria-nutrient
medium. The medium is changed after 14 days. Similarly another set of three cut cement mortar
specimens containing no filling were kept exposed to air. All the three sets of cement mortar
specimens were tested for the compressive strengths after 28 days.

(a) No filling in cut

(b) with filling in cut
Fig 9.1: Simulated standard cracks made in cement mortar cubes
(b) Realistic cracks
Realistic cracks were obtained in cement mortar specimens by applying at least 60 % of ultimate
load until a crack was visible with the naked eye. Then the cracked samples were placed in
bacteria Bacillus subtilis JC3 and nutrient medium for 28 days. During the period of immersion,
bacteria started to precipitate CaCO3 resulting in a compete filling of the crack. The deposition of
calcium carbonate was visually monitored periodically. At the end of the 28 day exposure, the
cubes were tested for compressive strength.
Test results and Discussion
The table 9.1 tabulates the compressive strength values of all three sets of cement mortar
specimens with standardized cracks.
Table Compressive strengths of cement mortar specimens with standard simulated cracks
Compressive strength
Specimen Type
at 28 days
(MPa)
Control specimen
52.6
(with no crack made)
Specimen with simulated Crack
28.91
(crack not filled up)
Specimen with simulated crack (crack filled up
34.56
with standard sand mixed with distilled water)
Specimen with simulated crack (crack filled up
with standard sand
mixed with bacterial 42.36
culture)

The above studies showed that the simulated cracked cube filled up with standard sand mixed
with bacterial culture has regained the strength of about 46%, when compared to the cut and non
remediated cube (cut with no fill up). Strength loss due to simulated crack left untreated (no fill

up) is 45%. Strength loss due to simulated crack treated with standard sand mixed with distilled
water is 34.3%. Strength loss due to simulated crack filled with standard sand mixed with
bacterial culture is 19.5 %. Strength gain due to biogenic treatment is 22.6% this is mainly due to
chemical bonding between CaCO3 precipitated by bacterial cells and sand particles which
consolidate the crack space.
Table Compressive strengths of cement mortar specimens with realistic cracks
Type of specimen
Compressive strength at 28 days (MPa)
Control cement mortar specimen
51.68
Cracked cement mortar specimen
44.99
(immersed in water)
Cracked cement mortar specimen
(immersed in bacterial culture)
49.17
(Bio-remediated)
Strength loss for cracked cement mortar specimens immersed in water is 13 %. Strength
loss for cracked cement mortar specimens treated in bacterial culture is 4.9 %. Strength gain due
to biogenic treatment is 9.3 %
Conclusions
The visual examination of crack surface of the cement mortar samples reveals the fully grown
calcite crystals, with distinct and sharp edges all over the surface of the crack, acts as an agent
for an eventual plugging and crack remediation. Bacteriogenic mineral precipitation contributed
to the bonding and regaining strength of the already cracked cubes. This microbial mixture with
sand filled in the cracks was found to remain intact after five days treatment confirming the
microbial calcite precipitation. Higher strength regain was obtained because of the bacterial
CaCO3 precipitation inside the simulated cracks of cement mortar specimens. This strength
recovery can be attributed to chemical bonding between CaCO3 precipitated by bacterial cells
and sand particles which consolidate the crack space.
Preliminary Cost Evaluation
The cost/benefit analysis of bacterial concrete balances the increased cost of the concrete against
substantial repair material costs, enhanced durability and aesthetic benefits. The benefits are
apparent at strength and performance of the finished product. Only expensive component in the
development of bacterial concrete is nutrients. In the market bacteria is available is lyophilized
state. So cost depends on the surface treatment area or volume of concrete used. Nutrients used
for this study are laboratory nutrients which are quite expensive so other inexpensive nutrient
sources can also be tried to reduce the commercial production cost of bacterial concrete.
However, any 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. These industrial effluents which are
potential environmental pollutants and also available locally with a price of nearly Rs 100 per
liter, which is very economic compared with standard laboratory nutrient medium. In this
project, to prepare one liter of nutrients mixed bacterial culture, it required 13 grams of nutrients
broth powder. The cost of 500 grams of HIMEDIA M002 nutrient broth powder costs about 2300

rupees so to prepare one litre of nutrients mixed bacterial culture costs Rs 60. In this project
nearly 125 liters of nutrients mixed bacterial culture was used costing nearly 7500 rupees.

[48] S.S. Bang, J.K. Galinat, and V. Ramakrishnan, Calcite precipitation

induced by polyurethane-immobilized Bacillus pasteurii,
Enzyme Microb. Tech. 28, 404409 (2001).
[49] W. De Muynck, D. Debrouwer, N. De Belie, and W. Verstraete,
Bacterial carbonate precipitation improves the durability
of cementitious materials, Cem. Concr. Res. 38, 1005
1014 (2008).
[50] H.M. Jonkers, Self healing concrete: a biological approach,
in Self Healing Materials an Alternative Approach to 20 Centuries
of Materials Science pp. 195204, ed. S. van der Zwaag,
Springer, Dordrecht, 2007.
[51] H.M. Jonkers and E. Schlangen, Development of a bacteriabased
self healing concrete, Proc. Int. FIB Symposium 1, 425
430 (2008).
[52] H.M. Jonkers and E. Schlangen, A two component bacteria
based self healing concrete, Concr. Repair, Rehab. and Retrofit.
1, 119120 (2009).
J. Aizenberg and P. Fratzl, Biological and biomimetic materials,
Adv. Mater. 21, 387388 (2009).

Bio-data of Authors
Author 1:
Dr M V Seshagiri Rao , FIE
(FIE No: 015739/9)
Professor in Civil Engineering
JNTUH College of Engineering
Hyderbad-85
Email: rao_vs_meduri@yahoo.com
Ph: 944 036 1817
Awards/ medals received:
1. Brij Mohan Lal best paper Award for the paper published in Vol.80; August 1999, IEI Journal
2. Best paper award for the paper presented at the 67th Annual technical session of
The Institution of Engineers awarded in October 2005.
3. Out standing Concrete technologist 2006 Award by Indian Concrete Institute(AP, Hyderabad )
4. Best paper award for the paper presented at the 70th Annual technical session of

The Institution of Engineers awarded in October 2008.

5. Best Teacher Award for the year 2009 by the Govt. of Andhra Pradesh
Research Publications

Memberships

175

Teaching & Research Experience: 38 yrs

FIE (Life Member), MISTE (Life Member), MICI (Life Member)

M.I.W.R.S (life Member)
Number of PhD Scholars Guided: 18
Author 2 :
Mr. V Srinivasa Reddy, MIE (M-146335-1)
Associate Professor in Civil Engineering
Gokaraju Rangaraju Institute of Engineering and Technology
Email: vempada@gmail.com
Ph: 970 468 3149
Research Publications

Memberships

50
MIE (Life Member), MISTE (Life Member), MICI (Life Member)