J Ind Microbiol Biotechnol (2017) 44:1511–1525

DOI 10.1007/s10295-017-1978-0

ENVIRONMENTAL MICROBIOLOGY - REVIEW

Microbial healing of cracks in concrete: a review

Sumit Joshi1 · Shweta Goyal2 · Abhijit Mukherjee3 · M. Sudhakara Reddy1 

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Received: 20 April 2017 / Accepted: 5 September 2017 / Published online: 12 September 2017
© Society for Industrial Microbiology and Biotechnology 2017

Abstract  Concrete is the most widely used construction Introduction

material of the world and maintaining concrete structures
from premature deterioration is proving to be a great chal- Concrete and steel are the two most commonly used struc-
lenge. Early age formation of micro-cracking in concrete tural materials. Among them, reinforced concrete is the most
structure severely affects the serviceability leading to high versatile and potentially one of the most durable materials
cost of maintenance. Apart from conventional methods of for almost any types of building structures. The factors that
repairing cracks with sealants or treating the concrete with affect the lifecycle performance of concrete include climate
adhesive chemicals to prevent the cracks from widening, change, higher temperature and extreme weather events
a microbial crack-healing approach has shown promising which lead to its premature deterioration. Most of these
results. The unique feature of the microbial system is that it threatening factors involve chemical attack on concrete or
enables self-healing of concrete. The effectiveness of micro- on the rebar embedded in it. The interconnected pore sys-
bially induced calcium carbonate precipitation (MICCP) tem and presence of micro-cracks also allow penetration of
in improving durability of cementitious building materi- harmful agents like ­CO2, ­SO42− and ­Cl− to cause corrosion
als, restoration of stone monuments and soil bioclogging of steel rebar [13]. The rate of deterioration of concrete
is discussed. Main emphasis has been laid on the potential structures mainly depends on two important factors: perme-
of bacteria-based crack repair in concrete structure and the ability of concrete matrix and development of micro-cracks
applications of different bacterial treatments to self-heal- [14]. As nearly 80% of world’s infrastructure is built in rein-
ing cracks. Furthermore, recommendations to employ the forced concrete, their maintenance needs a huge recurring
MICCP technology at commercial scale and reduction in the investment that few countries in the world can afford. There
cost of application are provided in this review. is a worldwide effort for sustainable technologies for main-
tenance of infrastructures that would offer economy without
Keywords  Microbial concrete · Autogenous healing · undue environmental or social costs.
Self-healing · Crack healing · Urea hydrolysis · Bacillus To minimize the cost of repair and rehabilitation of large-
scale civil infrastructure facing the deterioration from the
cracks, different strategies for the repair of cracks has been
studied. Intensive research to heal the concrete structures
suffering from cracks has been done by using organic and
* M. Sudhakara Reddy inorganic filling materials [90]. More recently, introduction
[email protected]
of applied biotechnology in the field of concrete has led to
1
Department of Biotechnology, Thapar University, Patiala, the development of a new domain called “microbial con-
Punjab 147004, India crete” or “bio-concrete”. It is a microbial-based strategy in
2
Department of Civil Engineering, Thapar University, Patiala, which concrete structures are treated with bacteria to induce
Punjab 147004, India calcium carbonate precipitation [4]. A positive benefit of
3
Department of Civil Engineering, Curtin University, biomineralization activity of various bacterial strains to
Bentley 6102, Australia enhance the durability and crack healing of concrete has

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1512 J Ind Microbiol Biotechnol (2017) 44:1511–1525

become an important research topic during the last decade. concrete was also cited as a probable healing mechanism.
Many reviews have been published addressing the applica- Swelling and hydration of cement paste, blocking of flow
tions of microbial concrete in various fields including the path by water impurities or by concrete particles broken
durability enhancement of building material [5, 10, 24, 26, from crack surface, and precipitation of calcium carbonate
28, 60, 68, 82, 87, 94]. The ecological benefits of MICCP crystals were suggested as possible chemical and physical
in urban development have been highlighted [5]. Different mechanisms contributing to the autogenous healing [37].
applications of the technology have been explored [10, 26, Formation of calcite in the crack seems to be the sole cause
60]. Different microbial pathways and their applicability for the autogenous healing and the crystal growth rate is
in construction have been analyzed [28, 94]. The sustain- dependent on the width of crack and water pressure, whereas
ability of this technology has been examined [82]. More concrete composition and water hardness have no influence
recently, the self-healing ability of the technology has been on autogenous healing [32]. The water entering the cracks
compared with the existing abiotic processes [87]. Self-heal- exhibits the pH value (5.5–7.5), ­CO2 content and certain

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ing is essentially autogenous filling of cracks that develop amount of ­Ca2+. As the ­CO2-containing water penetrates
in concrete. Its potential to make concrete sustainable has the hardened cement, it dissolves additional ­Ca2+ ions from
been explored [67]. However, there has been a rapid devel- Ca(OH)2 and calcium silicate hydrate phase of cement
opment in the understanding of causes of concrete cracking resulting in a rise in the pH of water. At pH >8 conversion of
and microbial techniques for healing the cracks. These are in-water bicarbonates into carbonates and C ­ a2+ ion concen-
the precursors for the development of self-healing concrete. tration favor precipitation of calcite. Reinhardt and Joss [63]
This paper presents a comprehensive review of the concrete reported that higher temperature favors a faster self-healing
cracking problems, different techniques for healing them while cracks ≤0.10 mm can be closed by self-healing. The
and finally, autogenous healing of concrete by the microbial main limitation of autogenous healing is the maximum crack
means. The mechanisms for microbially induced C ­ aCO3 width that can be healed is rather limited (~0.1 mm). Con-
precipitation and its applications in concrete structures are crete structures suffer from cracks that are much wider than
discussed. The efficacy of microbial techniques of healing that. Thus, engineered self-healing is essential.
them has been examined. Finally, a future direction for the
development of self-healing concrete is indicated. Engineered self‑healing in concrete

Supplementation of concrete with healing agents to stimulate

Approach in crack healing the healing action to repair cracks is known as engineered
self-healing. Many researchers used different healing agents
In the context of civil engineering research, healing is the and proposed their mechanisms on cementitious materials
phenomenon of restoration of concrete structure from a state (Table 1). Three different approaches are used to incorporate
of damage. In case of crack healing, chemical and physical self-healing material in concrete matrix and which activates
processes result in the introduction of secondary products its healing property when the crack appears in the structure.
that block or seal the cracks. To some extent, concrete is Based on the mechanisms, engineered self-healing concrete
capable of healing cracks autogenously, which is called self- are classified as vascular based, capsule based and intrinsic
healing or autogenous healing [39]. Engineered healing is based [15]. In vascular-based self-healing mechanism, heal-
warranted when autogenous healing is not adequate. Engi- ing material filled in hollow channels or fibers embedded in
neered self-healing is a process in which new substitute is concrete matrix are released when damage ruptures the hol-
incorporated into the cement material which is stimulated low channels or fibers (Fig. 1a). The healing agent stored in
on crack propagation resulting in sealing of crack [54, 87]. capsules is triggered through the release of healing material
in concrete matrix when capsules are ruptured by damage
Autogenous healing in concrete (Fig. 1b). Intrinsic self-healing materials embedded directly
in concrete matrix possess a latent self-healing functional-
Autogenous healing is a natural process of crack repair ity which is triggered by damage or by external stimulus
that occur in concrete in the presence of moisture and the (Fig. 1c) These materials rely on polymerizations, melting
absence of tensile stress. The repair is by a combination of of thermoplastic phases or ionic interactions to initiate self-
mechanical blocking by particles carried into the crack with healing. In the domain of engineered self-healing concrete,
the water and the deposition of calcium carbonate from the incorporation of hollow glass tubes filled with adhesive
cementitious material. Autogenous healing of cracks was chemicals in cement matrices was reported in earlier stud-
reported due to the formation of calcium carbonate pre- ies [31]. Use of polymer-based admixture was also reported
cipitation as consequence of water leakage through cracks to develop polymer modifying concrete or polymer modify-
[21]. Formation of calcite under wet conditions in cracks of ing mortar [57]. Application of different polymer modifiers

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J Ind Microbiol Biotechnol (2017) 44:1511–1525 1513
like acrylic, styrene- butadiene latex, polyvinyl acetate and
References

ethylene vinyl acetate in concrete crack sealing was reported

by Fowler [34]. Studies based on engineered self-healing
[31]

[57]

[40]

[66]

[71]
[9]

concrete investigated by researchers were epoxy repairing

of cracks by injection method and gravity filling method
Reduction in permeability and enhancement in flexural toughen-

Crack width of 0.2 mm was completely healed at 33-day curing

Surface crack introduced in specimen was submerged in water.

due to swelling effect, expansion effect and re-crystallization
Micro-cracks under stress in modified concrete are bridged by

[40], incorporation of high volume fly ash [66], expansive

Increase in tensile and compressive strength across a crack in

Mechanical and permeation properties of cracked specimens

­CaCO3 precipitation healed the surface crack in specimen

healing material, i.e., sodium aluminum silicate hydroxide

containing swelling agent montmorillonite [9], crystalline
additive and calcium sulfoaluminate-based expansive addi-
tive [71]. In aforementioned investigations, promising results
polymer films and prevent crack propagation
ing in active and passive mode, respectively

recovered on self-healing action of fly ash

in crack closure based on self-healing action were observed.

This technology has some limitations depending on the heal-
ing agents and strategies applied. In order to select effective Downloaded from https://academic.oup.com/jimb/article/44/11/1511/5996420 by guest on 10 November 2021
and reliable self-healing, care should be taken in choosing
a healing agent and a suitable strategy for application [87].
epoxy repair was observed

Application of microbially induced carbonate precipitation

Crack-healing outcomes

(MICCP) in civil engineering has become a topic of research

worldwide and this technology has been studied mainly for
application in the fields of surface protection of building
materials, crack remediation in concrete and soil improve-
Table 1  Overview of different healing agents and mechanism of healing action in cementitious matrix used by researchers

ment [28]. Adopting MICCP to fill the cracks is very innova-

tive and is pollution free and natural.
Expansive additive (calcium sulfoaluminate) 10% additive by mass of total cementitious material was admixed
Polymer latexes are used as admixtures in mortar and concrete or

Load-induced micro-cracks created in specimens were stored in

Incorporation of hollow porous polypropylene fibers filled with

Artificial cracks installed during casting in concrete cube were

Microbially induced calcium carbonate precipitation

Healing agents were used as admixture substitute in concrete
Fly ash in ratios of 35 and 55% by mass of binder was added

(MICCP)
repaired with epoxy resin by gravity filling method

MICCP is the capability of microbes to form calcium car-

bonate extracellularly through a metabolic activity. The
phenomenon of mineral formation by living organism due
specimens with cracks created artificially

to reaction of its metabolic products with the surrounding

environment is called biomineralization. The property of
healing agent in concrete structure

mineral formation by different bacterial species like sulfate-

reducing bacteria (SRB), silicate associated bacteria, unicel-
as grout repairing material
Mechanism of application

lular cyanobacteria and urea degrading bacteria has been

lime water for 30 days

reported [30]. Hammes and Verstraete [38] reported four key

factors such as (1) the calcium concentration; (2) the con-
centration of dissolved inorganic carbon (DIC); (3) the pH
and (4) the availability of nucleation sites which influence
the precipitation of ­CaCO3. Sufficient calcium and carbonate
ions are required for the ­CaCO3 precipitation so that the ion
activity product (IAP) exceeds the solubility constant (Kso)
Polymer latex-modified mortar or concrete

as in Eqs. 1 and 2. From the comparison of the IAP with the

Expansive agent ­(C4A3S, ­CaSO4, CaO)
Kso the saturation state (Ω) of the system can be defined; if

Chemical agent (­ NaHCO3, ­Na2CO3)

Ω > 1 the system is oversaturated and precipitation is likely

Geo-materials (montmorillonite)
as [55]:
Sikadur-52 (epoxy adhesive)
Ca2+ + CO32− ↔ CaCO3 (1)
Methyl methacrylate
𝛺 = 𝛼(Ca2+ )𝛼(CO32− )∕Kso with Kso calcite,25◦ = 4.8 × 10−9 .

Low-lime fly ash

(2)
The concentration of carbonate ions is related to the con-
centration of DIC and the pH of a given aquatic system. Also
Agent
the concentration of DIC depends on several environmental
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1514 J Ind Microbiol Biotechnol (2017) 44:1511–1525

Fig. 1  Different approaches in
engineered self-healing con-
crete. a In vascular-based self-
healing, hollow channels filled
with healing agent ruptures on
damage and releases healing
material. b In capsule-based
self-healing, healing agent is
released from ruptured capsules
on damage. c In intrinsic based
approach, healing agent possess
latent self-healing functionality
which is triggered on damage or
by external stimulus

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parameters such as temperature and the partial pressure of Heterotrophic‑mediated pathway
carbon dioxide (for systems exposed to atmosphere). The
equilibrium reactions and the constants governing the dis- In heterotrophic-mediated pathway, carbonate precipitation
solution of C
­ O2 in aqueous media (25 °C and 1 atm) are occurs either by sulphur cycle or by nitrogen cycle [18]. Sul-
given in Eqs. 3, 4, 5 and 6 [73]: phur cycle is carried out by sulphate reducing bacteria (SRB)
via dissimilatory reduction of sulphate. In this process, envi-
CO2(g) ↔ CO2(aq.) (pKH = 1.468), (3) ronment must be anoxic and rich in organic matter, calcium
CO2(aq.) + H2 O ↔ H2 CO∗3 (pK = 2.84), (4) and sulphate. As the organic matter is degraded by SRB, bicar-
bonate ions and hydrogen sulphide are produced by bacterial
H2 CO∗3 ↔ H+ + HCO−3 (pK1 = 6.352), (5) action using ­SO42− as a terminal electron acceptor as shown in
Eq. 7 [38, 56]. Elevation of pH in the surrounding environment
HCO−3 ↔ CO2−
3
+ H+ (pK2 = 10.329), (6) due to degasification of hydrogen sulphide in the presence of
with H2 CO∗3= CO2(aq.) + H2 CO3 Broadly two different ­Ca2+ induces the precipitation of calcium carbonate. In natural
metabolic pathways are involved in the process of biomin- environment if anoxygenic sulphide-phototrophic bacteria are
eralization associated with the microorganisms: (1) auto- present, hydrogen sulphide is used by bacteria and anaerobi-
trophic pathway and (2) heterotrophic pathway. cally oxidized to sulphur. Uptake of hydrogen sulphide results
in pH elevation and favors calcium carbonate precipitation.
Peckmann et al. [58] reported the precipitation of aragonite
Autotrophic‑mediated pathways crystals on dissolution of gypsum by the action of sulphate
reducing bacteria. They reported that gypsum present in cavi-
In autotrophic-mediated pathways, calcium carbonate pre- ties provides the calcium ions for aragonite precipitation and
cipitation is induced by microbes with the conversion of sulfate ions for the metabolic processes of sulphate reducing
carbon dioxide in the presence of calcium ions in its imme- bacteria as shown in Eq. 8. Degradation of organic matter in
diate environment. Autotrophic precipitation of carbonates anaerobic conditions provides the increased alkaline condition
includes non-methylotrophic methanogenesis, anoxygenic and facilitates the formation of aragonite crystals.
photosynthesis and oxygenic photosynthesis [17, 18, 36, 68]. 2CH2 O + SO2− → H2 S + 2HCO−3 (7)
4
All of the three autotrophic pathways use carbon dioxide as
a carbon source. CaSO4 .2H2 O → Ca2+ + SO2− + 2H2 O. (8)
4

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J Ind Microbiol Biotechnol (2017) 44:1511–1525 1515

Calcium carbonate precipitation by nitrogen cycle is positively charged cations (e.g., ­Ca2+, ­Mg2+) on the cell sur-
further categorized into three different mechanisms: (1) face. In bacteria, negatively charged groups dominate over
ammonification of amino acids (presence of organic mat- positively charged ones, giving the cell surface an overall
ter and calcium in aerobic conditions); (2) dissimilatory anionic charge resulting the deposition of divalent positively
reduction of nitrate (presence of organic matter, calcium charged metal ions on interaction [30]. Bacterial cell surface
and nitrate in anaerobic conditions) and (3) urea degrada- plays an important role in precipitation of calcium carbonate
tion (presence of organic matter, calcium and urea in aerobic as nucleation site as shown in Eqs. 14, 15 and 16:
conditions) [17]. In all these mechanisms, carbonate and
bicarbonate ions as well as ammonia ­(NH3) are produced Ca2+ + Cell ⟶ Cell Ca2+ , (14)
as a metabolic end product. Generation of ammonia creates Cl− + HCO−3 + NH3 ⟶ NH4 Cl + CO2− , (15)
high alkaline pH in the microenvironment of the bacterial
3

cell and decreased H ­ + concentration, affecting the carbon- Cell − Ca2+ + CO2− ⟶ Cell CaCO3 . (16)
3

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ate–bicarbonate equilibria shift towards the production of
­CO32− ions. Presence of calcium ions in the surrounding
of bacterial cell results in the precipitation of calcium car- Applications of MICCP in civil engineering
bonate. Among the aforementioned mechanisms in hetero-
trophic pathway, microbially induced calcium carbonate The potential and effectiveness of MICCP technology by
precipitation via urea hydrolysis is widely used in various using bacteria have been used widely in various fields. Sev-
applications. eral authors have reported the potential of MICCP property
in the application of remediation of heavy metal and radio-
nuclide contaminated ground water [35, 83], soil bioclog-
MICCP via urea hydrolysis ging [55, 75], restoration of stone monuments [22, 44, 64,
74] and durability enhancement of concrete structures [4, 6,
Microbially induced calcium carbonate precipitation via 16, 25, 27, 47, 62]. To reduce the permeability and increase
urea hydrolysis is an easily controlled mechanism in which the shear strength of soil, MICCP was performed by using
high amounts of carbonates are produced by the ureolytic Bacillus sp. with urea and calcium solution by Chu et al.
bacteria in short time period. In this mechanism, degradation [20]. Martinez et al. [53] optimized the MICCP process
of urea is catalyzed by microbial urease enzyme into carbon- for soil improvement by injecting Sporosarcina pasteurii
ate and ammonium [72]. One mole of urea is hydrolyzed in half-meter sand column in one-dimensional flow. Their
intracellularly to 1 mol of ammonia and 1 mol of carbamate study confirms that the most important factor for achieving
which spontaneously hydrolyses to form 1 mol of ammonia uniform calcium carbonate precipitation is the distribution
and carbonic acid as shown in Eqs. 9 and 10: of microbes. Stocks-Fischer et al. [72] reported the urease
activity of alkaliphilic bacteria to hydrolyze urea and high
Bacteria
CO(NH2 )2 + H2 ONH2 �������������→
� COOH + NH3 (9) pH being the favorable condition for calcite precipitation in
porous sand media.
To prevent the deterioration of monumental stones due
NH2 COOH + H2 O ⟶ NH3 + H2 CO3 . (10)
to weathering action, application of MICCP as a conserva-
These products further equilibrate in water to form
tive treatment has been reported by various researchers. Le
bicarbonate and 2 mol of ammonium and hydroxide ions
Metayer-Levrel [51] reported the reduction in water absorp-
(Eqs. 11, 12).
tion of limestone samples due to effective biocalcin coat-
2NH3 + 2H2 O ⟷ 2NH+4 + 2OH− (11) ing by carbonatogenic bacteria. Tiano et al. [74] reported
that limestone samples treated with Micrococcus sp., and
2OH− + H2 CO3 ⟷ CO2−
3
+ 2H2 O. (12) Bacillus subtilis by brushing showed a reduction of about
Generation of ammonia on the hydrolysis of urea results 60% in water absorption. Precipitation of calcite and vaterite
in pH increase creating an alkaline condition in the micro- crystals by Myxococcus xanthus resulted in efficient protec-
environment around the bacterial cell [72]. Presence of cal- tion and consolidation in porous limestone [64]. Daskalakis
cium ions in the surroundings of bacterial cell wall results in et al. [22] reported effective vaterite precipitation on the
the precipitation of calcium carbonate as the super-saturation marble sample treated with Bacillus pumilus strain isolated
is reached as shown in Eq. 13: from a cave stone. Improved surface drilling resistance and
reduction in water absorption was reported due to effective
CO2−
3
+ Ca2+ ⟷ CaCO3 . (13) biocalcification on Globigerina limestone specimen treated
The heterogeneous electronegatively charged bacterial with B. subtilis by spraying and poulticing [52].
cell wall acts as a nucleating site favoring the adsorption of

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1516 J Ind Microbiol Biotechnol (2017) 44:1511–1525

The potential of MICCP application in cementitious mate- of damage to concrete materials [26]. To overcome the
rials to enhance the mechanical properties as well as perme- problems associated with urea-based microbial carbonate
ability properties has been reported by various researchers precipitation, Zhu et al. [96] proposed an alternative technol-
as shown in Table 2. Precipitation of calcium carbonate by ogy using autophototrophic bacteria. Their studies showed
bacteria inside the cement matrix leads into pore refinement that biomineralization of cyanobacteria Synechococcus
resulting in reduced permeability and increased compres- PCC8806 formed a thick calcite-cell aggregate layer adher-
sive strength of concrete structure. Ramachandran et al. [62] ing to the concrete and decreased the water absorption and
reported increased compressive strength of cement mortar resistant to sonication. We proposed utilization of ­CO2 as
cubes on direct incorporation of live bacterial cells of S. alternative source to urea in biocementation [46]. Urea was
pasteurii strain inside the cement matrix. De Muynck et al. replaced with direct influx of C­ O2 and studied the precipita-
[25] reported the effectiveness of pure and mixed ureolytic tion of carbonates by Bacillus megaterium SS3. The bacteria
cultures in biodeposition on surface treatment of concrete. was able to grow well and precipitate carbonates with C ­ O2

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They observed that the use of pure culture of B. sphaericus influx and the amount of C ­ aCO3 precipitated is comparable
resulted in more pronounced decrease in uptake of water with that of urea used for precipitation. Our results showed
and gas permeability than the use of mixed ureolytic cul- 117 and 47% improvement in compressive strength with
ture. Achal et al. [6] reported the fly ash-amended mortar respect to control and with urea-treated specimens and sig-
and concrete specimens on treatment with ureolytic bacte- nificant reduction in water absorption in concrete specimens
rial strain B. megaterium showed improvement in strength [46]. Yeast extract often has been used as a carbon source in
and permeability properties. In our earlier reports, we have microbial induced precipitation of ­CaCO3. Severe retarda-
reported an increase in compressive strength of cement mor- tion of hydration kinetics has been observed due to the addi-
tar specimen and energy-efficient green building materials tion of yeast extract in cement. Williams et al. [86] replaced
by treating with bacteria [4, 27]. Kim et al. [47] investi- yeast extract with a combination of meat extract and sodium
gated the distribution of calcium carbonate precipitation acetate in growth medium for S. pasteurii. Their results sug-
and capillary water absorption of concrete specimens after gested that the medium reduced 75% retardation compared
surface treatment with two bacterial strains, B. sphaericus to yeast extract without compromising the growth, urea
and S. pasteurii individually. Denser calcium carbonate hydrolysis, cell zeta potential, and C­ aCO3 formation. Zhu
crystals and lowest weight increase was reported in speci- et al. [95] studied the MICCP by live and UV-killed pho-
mens treated with B. sphaericus strain than the specimens totrophic cyanobacteria Gloeocapsa PCC73106 in mortar
treated with S. pasteurii strain. Bundur et al. [16] reported specimens to enhance the durability properties. Their results
increased compressive strength of mortar specimen prepared suggested that treatments with live cells under illumination
with incorporation of vegetative bacterial cells than the con- increased the amount of precipitates, while UV-killed cells
trol specimen. Application of halophilic bacteria Exiguobac- increased the compressive strength, reduced water absorp-
terium mexicanum isolated from sea water showed 23.5% tion and lowest porosity.
increase in compressive strength and 5 times reduction in
water absorption on concrete specimens under 5% salt stress
condition [12]. Kumari et al. [50] reported 49% increase Bio‑inspired applications in crack healing
in compressive strength by using non-ureolytic bacteria
Bacillus cohnii. Our earlier studies showed that Bacillus sp. Formation of crack is a commonly observed phenomenon
CT-5-treated reinforced concrete (RC) specimens reduced in concrete structures. Though micro-crack formation may
the corrosion rate, reduction in mass loss and increase in hardly affect the structural properties of constructions,
pullout strength than the control specimens [7]. Kalhori increased permeability due to micro-crack networking sub-
and Bagherpour [45] studied the effect of ­CaCO3 precipi- stantially reduces the durability of concrete structures due
tating bacteria Bacillus subtilis on healing and mechanical to risk of ingress of aggressive substances particularly in
properties of shotcrete. Their results showed 30% increase moist environments. In order to increase the often observed
in the compressive strength of bacteria-exposed shotcrete autogenous crack-healing potential of concrete, specific
specimens compared to control specimens. The presence of healing agents are incorporated into the concrete matrix.
bacteria in the mix design and curing solution enhanced the Apart from the surface application and incorporation of live
tensile strength, decreased the water absorption and porosity bacterial cells inside the cement matrix, use of bacteria as a
of shotcrete. self-healing agent is also evolved as new approach. Differ-
MICCP via urea hydrolysis by urease enzyme offers ent methodologies have been adopted by researchers to test
unlimited advantages in the field of carbonate precipita- the compatibility of calcifying bacterial strains to heal the
tion, but limitation to the process include ammonia produc- cracks autonomously (Table 3). Initially, the application of
tion, which have environmental concerns as well as risks bacteria to remediate the cracks in cement mortar specimens

13
 J Ind Microbiol Biotechnol (2017) 44:1511–1525
Table 2  Overview of applications of different microorganisms and nutrient media to prevent deterioration of monumental stones and cementitious materials with microbial C
­ aCO3 precipitation
Microorganism Nutrient Application Mechanism of treatment Evaluation of specimens References

Micrococcus sp. Bacillus B4 nutrient medium (cal- Monumental limestone con- Samples were brushed with bacteria Water absorption, colorimetric analysis, [74]
subtilis. cium acetate, yeast extract, servation and kept wet with ­B4 medium for stone cohesion, SEM, XRD and FTIR
dextrose) 15 days to feed the bacteria of crystals
Myxococcus xanthus Pancreatic digest of casein, Porous ornamental limestone Immersion of samples (shaking and Weight increase, MIP, XRD, SEM analy- [64]
calcium acetate, potassium stationary conditions) in bacterial sis, sonication analysis.
carbonate culture with nutrient medium for
30 days
M. xanthus M-3P nutrient solution Archaeological gypsum Bacterial solution was sprayed till Drilling resistance analysis, TGA, XRD, [44]
(pancreatic digest of casein, plasters 6 days (twice a day) on the upper SEM, MIP, TEM and colorimetric
calcium acetate, potassium surface of sample analysis
carbonate)
B. pumilus Basic growth medium (bacte- Marble substrate Marble samples were sprayed every Chromatic analysis, weight loss with [22]
riological peptone, calcium 12 h with bacterial culture for ultrasonic treatment, XRD, SEM and
acetate) 15 days FTIR
Sporosarcina pasteurii Urea and calcium chloride Porous sand column Sand slurry mixed with bacterial cells XRD, SEM, ­CaCO3 estimation [72]
medium and urea–CaCl2 medium was fed
with nutrient medium for 10 days
S. pasteurii Urea and calcium chloride Cement mortar Casting of mortar cubes with bacte- Compressive strength, XRD and SEM [62]
Pseudomonas aeruginosa medium rial cells and cured in urea–CaCl2 analysis
medium
B. sphaericus Urea, nutrient broth, calcium Mortar and concrete speci- Specimens were immersed in Compressive strength, sorptivity test, [25]
chloride and calcium acetate mens bacterial culture for 1 day prior to SEM, gas permeability, XRD, chro-
submersion in nutrient medium for matic analysis of specimens
6 days
B. megaterium Nutrient broth, urea and Mortar and concrete speci- NBU-bacteria and fly ash admixed Compressive strength, water imperme- [6]
calcium chloride mens specimens were prepared and cured ability test, water absorption test and
with respective medium for 28 days SEM analysis
Bacillus sp. CT-5 Nutrient broth, urea and Cement mortar Mortar mixture was admixed with Compressive strength, water absorption [4]
calcium chloride NBU medium-bacterial cells and test and SEM analysis
cured with respective medium for
28 days
B. megaterium SS3 Nutrient broth, urea and Cement mortar blocks and Bacterial-admixed specimens were Water absorption test, SEM–EDX, XRD, [27]
calcium chloride cylinders cured by spraying with respective MIP and C­ aCO3 estimation of treated
medium for 28 days samples
S. pasteurii Nutrient broth, urea and Concrete specimens Specimen’s top surface was wetted Water absorption test, SEM–EDX and [47]
B. sphaericus calcium acetate with nutrient medium containing XRD analysis
S. pasteurii and B. sphaericus cells
separately for 28 days
S. pasteurii Urea, yeast extract medium Cement mortar Vegetative cells with UYE medium Hydration kinetics, compressive strength, [17]
13

were added in mortar mixture while TGA and XRD analysis

casting and cured by submersion in
UYE medium with lime till 56 days

1517
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1518 J Ind Microbiol Biotechnol (2017) 44:1511–1525

was investigated by Ramachandran et al. [62]. Cracks in Sharma et al. [69] demonstrated the potential application of
cement mortar beams and cubes were simulated artificially alkaliphilic Bacillus pseudofirmus in concrete crack repair
with constant width 3.175 mm and different depths. Their by rapid spore production and germination, calcium carbon-
results suggested that calcite precipitated during microbial ate formation in vitro and in situ.
growth enhanced the compressive strength of cracked mortar The combination of non-ureolytic bacteria with organic
cubes. The mineralization process was effective in shallow calcium source as a two-component self-healing system was
cracks than in deeper ones because the bacteria grow more proposed by Jonkers et al. [43]. The organic calcium com-
actively in presence of oxygen. To enhance the effective- pounds are converted to calcium carbonate due to respiration
ness of MICCP in remediation of deep cracks, polyurethane- effect of bacteria. They reported that by this method true
immobilized S. pasteurii cells were used in cement matrices self-healing can be achieved because all the components are
[11]. In cement mortar cubes (50.8 × 50.8 × 50.8 mm) with added to the concrete mixture prior to casting and become
simulated cracks of width 3.18 and 25.4 mm crack depth, an integral part of the concrete. Wiktor and Jonkers [84]

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polyurethane strip encapsulating bacterial cells was placed incorporated a mixture of bacterial spores (Bacillus alka-
in the cracks. Precipitation of calcite throughout of the crack linitrilicus) and calcium lactate embedded in expanded clay
matrices was reported as the polyurethane matrix provides particles as self-healing agent in concrete. Multiple cracks of
protection to bacterial cells from the extreme alkaline nature widths ranging from 0.05 to 1.0 mm were created in speci-
of concrete. Jonkers et al. [42] used the spores of Bacil- men on stretching embedded steel by applying tensile force.
lus pseudofirmus than vegetative cells as self-healing agent Cracked bacterial embedded specimens and control speci-
in concrete matrix to seal the cracks. The healing effect in mens were immersed in water to investigate the self-healing
freshly formed cracks appears on revival of immobilized properties. It was reported that after 100 days of immersion
spores by entering water and growth nutrients. To augment in water, bacterial-based specimen showed crack-healing of
the potential of bacteria as a self-healing agent in crack fill- up to 0.46 mm while in control it was only up to 0.18 mm
ing, immobilization of bacterial cells with different carriers (Fig. 2). Xu and Yao [88] investigated non-ureolytic bacteri-
had been studied. The biochemical healing agent, consisting ally induced ­CaCO3 precipitation as a self-healing strategy
a mixture of viable but dormant bacteria and organic com- for concrete cracking by using Bacillus cohnii spores. They
pounds packed in porous expanded clay particles to heal the suggested that incorporation of bacteria and calcium source
cracks was proposed by Jonkers [41]. A two-dimensional nutrients as a two-component healing agent in concrete
mathematical model of bacterial crack healing was devel- matrix induces ­CaCO3 precipitation upon crack formation.
oped by Zemskov et al. [91] to estimate the influence of Crack width in range of 0.1–0.4 mm was sealed completely
different parameters involved on the rate and quality of the as well as a layer of precipitates on the surface of speci-
crack healing. To protect the bacteria from strong alkaline men was reported in externally applied healing. They also
environment of concrete, bacteria (B. sphaericus) immobi- reported that self-healing efficiency in specimens with incor-
lized in silica gel was investigated by Tittelboom et al. [75]. porated bacterial spores and nutritional agents was higher
Standard cracks of width 0.3 mm with two depths of 10 and than the control specimens. Higher efficiency of calcium
20 mm were prepared in concrete samples by introducing carbonate precipitation in crack healing on external treat-
thin copper plate in cement paste while casting. Realistic ment and self-healing application was observed in calcium
cracks with width range from 0.05 to 0.87 mm were also glutamate precursor than calcium lactate. However, Xu et al.
created in concrete cylinders of diameter 80 mm and height [89] reported a higher rate of ­CaCO3 precipitation with cal-
75 mm by subjecting to splitting test and used for water cium lactate than calcium nitrate in case of ureolytic bacteria
permeability test. Specimens treated with silica gel-immo- Sporosarcina pasteurii. Zhang et al. [92] reported that pres-
bilized bacteria showed promising results in crack filling and ence of excessive C ­ a2+ used in biomineralization not only
low water permeability similar to epoxy-treated specimens. inhibits the C­ aCO3 precipitation, but also results in waste of
Tziviloglou et al. [76] incorporated the bacteria-based the ­Ca2+ resource. They suggested that C ­ a2+ concentration
healing agent into lightweight aggregates and mixed with lower than 30 mM is a good strategy for biomineralization
fresh mortar and evaluated the recovery of liquid tightness process. A binary self-healing system consisting of oxygen
after cracking and exposure to two different healing regimes releasing tablet (ORT) and bacteria (Bacillus H4) was devel-
(water immersion and wet–dry cycles) through water perme- oped to heal the cracks [93]. The ORT contains ­CaO2 and
ability tests. Their results revealed that the recovery of water lactic acid (9:1) which provides a stable oxygen supply and
tightness does not differ substantially for specimens with or maintains pH (9.5–11.0) for effective metabolic activity.
without healing agent when immersed continuously in water. We earlier reported the remediation of simulated cracks in
However, the recovery of water tightness increases signifi- mortar specimens (width 3 mm and depths of 13.4, 18.8, and
cantly for specimens containing the healing agent compared 27.2 mm) by using bacterial strain Bacillus sp. CT-5 [8]. Our
to specimens without it, when subjected to wet–dry cycles. results showed an increase of 40 and 37% in compressive

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strength in bacterial-treated specimen with crack depth of with complete crack closing up to 0.45 mm crack width after
13.4 and 27.2 mm, respectively, as compared to control. 4 weeks. In specimen without CERUP, autogenous crack
Successful healing of deepest crack of depth 27.2 mm was healing was observed with closing of crack with 0.25 mm
reported in bacterial-treated specimen (Fig. 3). Qian et al. crack width (Fig. 5). Effective crack healing was performed
[61] reported the healing of early age cracks in cement-based by directly pooling the S. pasteurii culture supplemented
materials by carbonic anhydrase-producing bacteria Bacillus with ­CaCl2 and urea around the cracked area of fiber con-
mucilaginosus L3. Their experimental results showed that crete beam [65]. They reported about 100% consolidation
the cracks formed at early ages were completely healed (up of the micro-cracks to a depth of approximately 20 mm.
to 0.4 mm) due to bacterial treatment and the healing effect Silva et al. [70] analyzed the costs involved in biological
reduced with the increasing of cracking age. To protect bac- self-healing in concrete and emphasized to develop the bio-
teria from high pH environment of concrete, Wang et al. [78] additive at much lower costs to make the biological self-
used diatomaceous earth to immobilize the bacteria. healing industrially applicable.

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The immobilized bacteria healed the cracks with a width Apart from ureolytic pathway in MICCP, self-protected
ranging from 0.15 to 0.17 mm compared to control speci- nitrate ­(NO3−) reducing bacteria was used for repair of con-
mens. Wang et al. [80] encapsulated bacterial spores into crete cracks [33]. Nitrate reducing bacteria was grown in
hydrogel and then incorporated into specimens to investigate enrichment culture by using concrete admixtures such as
their healing efficiency. Hydrogel was used because of its Ca(NO3)2 and Ca(HCOO)2 as nutrients. The culture grown
water retention properties and providing favorable microen- as granules (0.5–2.0 mm) consisting of 70% biomass and
vironment to bacterial spores with moisture and nutrients for 30% inorganic salts was added into mortar without any
activation. ­CaCO3 precipitation by hydrogel-encapsulated additional protection. They reported that, more than 90%
spores was demonstrated by thermogravimetric analysis of the crack with 500 μm crack width was closed and 68%
(TGA). Their findings suggest that sufficient amount of less water was absorbed by microbial specimen than the
water is essential for the bacterial spores to achieve the real- reference specimen upon 28-day immersion in water. In
istic self-healing mechanism to seal the cracks. The mortar specimens cured for 6 months, in microbial specimen 90%
specimens with hydrogel-encapsulated spores healed crack crack closure was observed in cracks with crack width up to
width of about 0.5  mm and the water permeability was 400 μm and in reference specimen crack closure was 86% for
decreased by 68% compared to control where maximum 135 μm crack width. Their results suggested that enriched
healed crack width was 0–0.3 mm and the average water mixed denitrifying cultures structured in self-protecting
permeability was decreased only by 15–55% (Fig. 4). granules are very promising strategies to enhance microbial
Application of microcapsules as self-healing agent carrier self-healing.
in crack remediation was investigated by Wang et al. [81]. The field-based potential of MICCP in sealing of sand-
The spores of B. sphaericus were encapsulated in melamine- stone fractures below the ground surface in wellbore using
based microcapsules. They reported that the crack-healing ureolytic bacteria was demonstrated by Phillips et al. [59].
ratio was much higher for specimens treated with bacteria Horizontally fractured sandstone 340.8 m below the ground
compared to the ones without bacteria where 18–50% of surface was sealed with ureolytic bacterial strain S. pasteurii
crack area healed in control while it was 48–80% in the bac- using conventional oil field delivery technology. In the field
teria-treated specimens. The maximum crack width healed test, bacterial culture was injected into the fracture by using
in the bacterial-treated specimens was 970 μm which was an 11.4 l wireline dump bailer for 4 days. Microbial sus-
about 4 times wider than control (250 μm). Wang et al. [79] pensions after injection in fracture was allowed to attach
tested modified alginate-based hydrogel as encapsulating for 1 h and later on the inoculum was amended with cal-
agent for application of B. sphaericus spores in concrete. cium containing growth solution (urea and nutrient broth)
Their results indicated efficient protection of the hydrogel to promote the growth. In this treatment, 24 urea/calcium
for spores in concrete and great potential to be used for crack solution and 6 microbial suspensions were injected into the
self-healing in concrete applications. fracture zone for 4 days. They reported that the flow rate
To reduce the operational cost of bio-based self-healing decreased from 1.9 to 0.47 L/min and in-well pressure falloff
action in concrete, a new powderous material containing an from >30% before to 7% after treatment. Their findings sug-
efficient ureolytic microbial community (Cyclic EnRiched gest that MICCP is a promising tool for sealing subsurface
Ureolytic Powder or CERUP) has been developed [23]. fractures in the near wellbore environment. To monitor the
CERUP was produced from a sub-stream of a vegetable applications of MICCP nondestructively in time and space,
treatment plant containing non-axenic bacterial culture with Krikland et al. [48] used a low-field nuclear magnetic reso-
the ability to sporulate. After drying, it was ground to a nance (NMR) well-logging probe in a sand-filled bioreactor
particle size below 500 μm in diameter. Highest capacity of by inoculating S. pasteurii and pulsed injections of urea and
crack healing was observed in CERUP admixed specimen calcium substrate. NMR signal amplitude and T ­ 2 relaxation

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Table 3  Overview of crack remediation in concrete using bacteria as crack-healing agent

Microorganism Carrier material Incubation treatment Crack healing References

Bacillus pasteurii Cells mixed with sand After microbial plugging mortar Crack with depth of 3.175 mm [62]
cubes were immersed in urea–
CaCl2 medium for 28 days
Bacillus pasteurii Polyurethane-immobilized cells Incubated in urea–CaCl2 medium for Crack width of 3.18 mm and depth [11]
28 days of 25.4 mm
Bacillus sphaericus Cells immobilized in silica gel Immersed in solution of urea and Crack width of 0.3 mm and depths [75]
calcium source for 3 days of 10.0 and 20.0 mm
Bacillus alkalinitrilicus Spores embedded in expanded clay Immersed in water for 100 days Crack width ranging from 0.05 to [84]
with calcium lactate 1.0 mm
Bacillus sp. CT-5 Cells mixed with sand Immersed in urea and ­CaCl2 Crack width of 3.0 mm and depths [8]
medium for 28 days of 13.4, 18.8 and 27.2 mm
Bacillus sphaericus Hydrogel-encapsulated spores with Submerged in water for 4 weeks Crack width of 0.5 mm [80]
nutrient and calcium source with wet–dry cycle
Bacillus cohnii Treated externally Submerged in medium containing Crack width ranging 0.1–0.4 mm [88]
bacterial spores, yeast extract and
calcium source
Bacillus sphaericus Spores encapsulated in microcapsule Immersed in water for 8 weeks with Maximum crack width healed is [81]

J Ind Microbiol Biotechnol (2017) 44:1511–1525

wet–dry cycle 0.97 mm
Non-axenic ureolytic spores Cyclic enriched ureolytic powder Immersed in urea and de-mineral- Crack healing of width 0.45 mm [23]
ized water for 4 weeks
Bacillus sphaericus Spores encapsulated in modified Fully immersed in water NA [79]
alginate hydrogel

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J Ind Microbiol Biotechnol (2017) 44:1511–1525 1521

Fig. 2  Stereomicroscopic
images of before and after
crack-healing process in control
(a, c) and bacterial-treated (b,
d) mortar specimens, respec-
tively Reprinted from Wiktor
and Jonkers [84] with permis-
sion from Elsevier

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were measured after the experiments. They reported that that only germinate when they come into contact with the
the water content in the reactor decreased to 76% of its ini- pH of concrete. Upon germination, the cells would descend
tial value and changes in T ­ 2 relaxation distributions due into cracks in the concrete. The bacteria use quorum sens-
to changes in pore volume and surface mineralogy. Their ing to determine when sufficient bacteria have accumulated,
results indicate the low-field NMR well-logging probe is triggering production of a mixture of calcium carbonate and
sensitive to the physical and chemical changes caused by a “bacterial glue”, which combines with the bacterial cells
MICCP in a laboratory bioreactor. Wiktor and Jonkers [85] to fill the crack [29]. Chattopadhyay and Sarkar [19] trans-
studied field performance of bacteria-based repair system formed the gene having silica-leaching attribute to a Bacil-
of a two-story parking garage suffering from cracking and lus subtilis and the transformed bacterial cells are utilized
damaged concrete pavement due to freeze/thaw. Denitrify- directly for enhancing the strength and durability properties
ing bacteria supplemented with two solutions (1) Solution of concrete structures. We earlier reported the development
A (sodium silicate, sodium gluconate) and (2) Solution B of phenotypic mutant of Sporosarcina pasteurii by UV irra-
(calcium nitrate) were treated by spraying manually until diation, which enhanced the urease activity and also calcite
saturation of concrete. Crack-sealing efficiency was evalu- production [2].
ated after 2 months of bacteria-based application in treated
area of parking garage by means of water permeability
test. Concrete pavement was also evaluated in resistance to
freeze/thaw conditions and deicing salts by drilling cores Conclusion and future perspectives
from treated area. They reported that cracks that had not
been treated with the bacteria-based repair system were still Bacterial remediation of cracks has shown very promising
heavily leaking and freeze/thaw resistance of concrete was results. The main advantage of the bacterial system over the
higher with the bio-based repair system than the untreated conventional systems is the lower viscosity of the bacterial
concrete. Tziviloglou et al. [77] described the various steps cementation fluid. In case of cement grout or the polymeric
taken towards the outdoor applications of bio-based self- resin healing, the ingredients are premixed before pumping
healing, for the laboratory-scale tests which have shown into the cracks. As a result, the cementation reaction com-
promising results. mences before the ingredients have reached the site of heal-
The use of genetically modified microbes to heal the ing. While in case of the bacterial system, the bacteria are
cracks is another area of research to improve the durability pumped in first. The cementation solution is then pumped
properties of concrete. The “BacillaFilla” is a genetically where the ingredients are dissolved in water. The cemen-
modified version of a Bacillus subtilis which contains cells tation reaction commences when the ingredients come in

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Fig. 3  Microscopic image
of a remediated crack with b
enlarged portion of remediated
crack showing calcite precipita-
tion and c rod-shaped bacteria
Reprinted from Achal et al. [8]
with permission from Elsevier

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contact of the bacterial enzyme. Thus, bacterial healing is this technology in several cases. Successful commerciali-
able to penetrate deep into the cracks. However, promising zation of the technique requires economical alternatives of
results from the microbial-based application in crack heal- the medium ingredients that cost as high as 60% of the total
ing of concrete at lab scale have been reported. Researchers operating costs [49]. Use of inexpensive materials as nutri-
from all over the world have investigated the potential of ents may help to lower the cost of treatment [1, 3]. The other
biomineralization in crack remediation. The qualitative and limitation in this application is external treatment of cracks
quantitative evaluation of microbial application to heal the in existing concrete structures. Efficiency of bacteria in
cracks in concrete are still reported in the lab scale. Some ­CaCO3 precipitation in deeper parts of the crack on external
researchers have also reported encouraging results on the treatment has to be investigated. In field-scale experiments,
application of bacterial-based treatment at field scale. Some effective curing method is to be investigated to provide suf-
limitations which are necessary to be considered for applica- ficient amount of nutrients and water to the bacterial cells.
tion of this technology at commercial scale have been found. Further investigations are needed to improve the microbial
Use of laboratory grade nutrient sources in field applications technology as an innovative crack-healing application for
is one of the economic limitations which restrict the use of the commercial scale.

Fig. 4  Maximum crack width of 0.5 mm healed in the specimen treated with hydrogel-encapsulated bacterial spores Reprinted from Wang et al.
[80] with permission from Elsevier

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Fig. 5  Photomicrographs of cracked mortar specimens on day 0 and Ureolytic Powder (CERUP) on day 0 and c1, d1 on day 28 (1). e, f
on day 28. a, b Reference samples on day 0 and a1, b1 on day 28; Mortar samples containing 1% of CERUP on day 0 and e1, f1 on day
c, d mortar samples containing 1% of autoclaved Cyclic EnRiched 28 Reprinted from Da Silva et al. [23] with permission from Elsevier

Acknowledgements  Authors are thankful to Department of Science 7. Achal V, Mukherjee A, Goyal S, Reddy MS (2012) Corrosion
and Technology, Govt. of India, for providing financial support to carry prevention of reinforced concrete with microbial calcite precipita-
out research in “Durability enhancement and prevention of damages in tion. ACI Mater J 109:157–164
reinforced structures using bacteria (DST no: SB/S3/CEE/0063/2013)”. 8. Achal V, Mukerjee A, Reddy MS (2013) Biogenic treatment
improves the durability and remediates the cracks of concrete
structures. Constr Build Mater 48:1–5
9. Ahn TH, Kishi T (2010) Crack self-healing behavior of cementi-
tious composites incorporating various mineral admixtures. J Adv
References Concr Technol 8:171–186
10. Anbu P, Kang CH, Shin YJ, So JS (2016) Formations of cal-
1. Achal V, Mukherjee A, Basu PC, Reddy MS (2009) Lactose cium carbonate minerals by bacteria and its multiple applications.
mother liquor as an alternative nutrient source for microbial con- Springerplus 5:250
crete production by Sporosarcina pasteurii. J Ind Microbiol Bio- 11. Bang SS, Galinat JK, Ramakrishnan V (2001) Calcite precipi-
technol 36:433–438 tation induced by polyurethane-immobilized Bacillus pasteurii.
2. Achal V, Mukherjee A, Basu PC, Reddy MS (2009) Strain Enzyme Microb Technol 28:404–409
improvement of Sporosarcina pasteurii for enhanced urease and 12. Bansal R, Dhami NK, Mukherjee A, Reddy MS (2016) Biocalcifi-
calcite production. J Ind Microbiol Biotechnol 36:981–988 cation by halophilic bacteria for remediation of concrete structures
3. Achal V, Mukherjee A, Reddy MS (2010) Biocalcification by in marine environment. J Ind Microbiol Biotechnol 43:1497–1505
Sporosarcina pasteurii using corn steep liquor as nutrient source. 13. Basheer L, Kropp J, Cleland DJ (2001) Assessment of the durabil-
Ind Biotechnol 6:170–174 ity of concrete from its permeation properties: a review. Constr
4. Achal V, Mukherjee A, Reddy MS (2011) Microbial Concrete: Build Mater 15:93–103
way to enhance the durability of building structures. J Mater Civil 14. Basheer PAM, Chidiac SE, Long AE (1996) Predictive models for
Eng 23:730–734 deterioration of concrete structures. Constr Build Mater 10:27–37
5. Achal V, Mukherjee A, Zhang Q (2016) Unearthing ecological 15. Blaiszik BJ, Kramer SLB, Olugebefola SC, Moore JS, Sottos NR,
wisdom from natural habitats and its ramifications on develop- White SR (2010) Self-healing polymers and composites. Annu
ment of biocement and sustainable cities. Landsc Urban Plan Rev Mater Res 40:179–211
155:61–68 16. Bundur ZB, Kirisits MJ, Ferron RD (2015) Biomineralized
6. Achal V, Pan X, Özyurt N (2011) Improved strength and durability cement-based materials: impact of inoculating vegetative bacte-
of fly ash-amended concrete by microbial calcite precipitation. rial cells on hydration and strength. Cem Concr Res 67:237–245
Ecol Eng 37:554–559 17. Castanier S, Le Me´tayer-Levrel G, Perthuisot JP (2000) Bacterial
roles in the precipitation of carbonate minerals. Microbial sedi-
ments. Springer, Heidelberg, pp 32–39

13

1524 J Ind Microbiol Biotechnol (2017) 44:1511–1525

18. Castanier S, Le Me´tayer-Levrel G, Perthuisot JP (1999) Ca-car- 40. Issa CA, Debs P (2007) Experimental study of epoxy repairing of
bonates precipitation and limestone genesis—the microbiogeolo- cracks in concrete. Constr Build Mater 21:157–163
gist point of view. Sediment Geol 126:9–23 41. Jonkers HM (2011) Bacteria-based self healing concrete. Heron
19. Chattopadhyay B, Sarkar M (2016) Genetically modified Bacillus 56:1–12
subtilis bacterial strain for self-healing and sustainable green bio- 42. Jonkers HM, Schlangen E (2007) Crack repair by concrete-immo-
concrete material. Org Chem Curr Res 5:3 bilized bacteria. In: Proceedings of the first international confer-
20. Chu J, Stabnikov V, Ivanov V (2012) Microbially induced calcium ence on self-healing materials, pp 18–20
carbonate precipitation on surface or in the bulk of soil. Geomi- 43. Jonkers HM, Thijssen A, Muyzer G, Copuroglu O, Schlangen E
crobiol J 29:544–549 (2010) Application of bacteria as self-healing agent for the devel-
21. Clear CA (1985) The effect of autogenous healing upon leakage opment of sustainable concrete. Ecol Eng 36:230–235
of water through cracks in concrete. Cem Concr Assoc (Tech Rpt 44. Jroundi F, Gonzalez-Muñoz MT, Garcia-Bueno A, Rodriguez-
559) Navarro C (2014) Consolidation of archaeological gypsum plaster
22. Daskalakis MI, Rigas F, Bakolas A, Magoulas A, Kotoulas G, by bacterial biomineralization of calcium carbonate. Acta Bio-
Katsikis I, Karageorgis AP, Mavridou A (2015) Vaterite bio- mater 10:3844–3854
precipitation induced by Bacillus pumilus isolated from a solu- 45. Kalhori H, Bagherpour R (2017) Application of carbonate pre-

Downloaded from https://academic.oup.com/jimb/article/44/11/1511/5996420 by guest on 10 November 2021

tional cave in Paiania, Athens, Greece. Int Biodeterior Biodegrad cipitating bacteria for improving properties and repairing cracks
99:73–84 of shotcrete. Constr Build Mater 148:249–260
23. Da Silva FB, De Belie N, Boon N, Verstraete W (2015) Production 46. Kaur G, Dhami NK, Goyal S, Mukherjee A, Reddy MS (2016)
of non-axenic ureolytic spores for self-healing concrete applica- Utilization of carbon dioxide as an alternative to urea in bioce-
tions. Constr Build Mater 93:1034–1041 mentation. Constr Build Mater 123:527–533
24. De Belie N (2016) Application of bacteria in concrete: a critical 47. Kim HK, Park SJ, Han JI, Lee HK (2013) Microbially mediated
review. RILEM Tech Lett 1:56–61 calcium carbonate precipitation on normal and lightweight con-
25. De Muynck W, Cox K, De Belie N, Verstraete W (2008) Bacte- crete. Constr Build Mater 38:1073–1082
rial carbonate precipitation as an alternative surface treatment for 48. Kirkland CM, Zanetti S, Grunewald E, Walsh WO, Codd SL, Phil-
concrete. Constr Build Mater 22:875–885 lips AJ (2017) Detecting microbially induced calcite precipitation
26. De Muynck W, De Belie N, Verstraete W (2010) Microbial car- in a model well-bore using downhole low-field NMR. Environ Sci
bonate precipitation in construction materials: a review. Ecol Eng Technol 51:1537–1543
36:118–136 49. Kristiansen B (2001) Process economics. In: Ratledge C, Kristian-
27. Dhami NK, Reddy MS, Mukherjee A (2013) Bacillus megaterium sen B (eds) Biotechnology, 2nd edn. Cambridge University Press,
mediated mineralization of calcium carbonate as biogenic surface Cambridge
treatment of green building materials. World J Microbiol Biotech- 50. Kumari C, Das B, Jayabalan R, Davis R, Sarkar P (2017) Effect of
nol 29:2397–2406 nonureolytic bacteria on engineering properties of cement mortar.
28. Dhami NK, Reddy MS, Mukherjee A (2013) Biomineralization J Mater Civ Eng 29:06016024
of calcium carbonates and their engineered applications: a review. 51. Le Metayer-Levrel G, Castanier S, Orial G, Loubiere JF, Perthui-
Front Microbiol 4:314 sot JP (1999) Applications of bacterial carbonatogenesis to the
29. Dillow C (2010) Engineered bacteria can fill cracks in aging con- protection and regeneration of limestones in buildings and historic
crete. In: Popular Science. Available via DIALOG. http://www. patrimony. Sediment Geol 126:25–34
popsci.com/science/article/2010-11/modified-bacterial-glue-can- 52. Micallef R, Vella D, Sinagra E, Zammit G (2016) Biocalcifying
shore-cracking-concrete. Accessed 14 Jul 2017 Bacillus subtilis cells effectively consolidate deteriorated Globige-
30. Douglas S, Beveridge TJ (1998) Mineral formation by bacteria in rina limestone. J Ind Microbiol Biotechnol 43:941–952
natural microbial communities. FEMS Microbiol Ecol 26:79–88 53. Martinez BC, DeJong JT, Ginn TR, Montoya BM, Barkouki TH,
31. Dry C (1994) Matrix cracking repair and filling using active and Hunt C, Tanyu B, Major D (2013) Experimental optimization of
passive modes for smart timed release of chemicals from fibres microbial-induced carbonate precipitation for soil improvement.
into cement matrices. Smart Mater Struct 3:118–123 J Geotech Geoenviron 139:587–598
32. Edvardsen C (1999) Water permeability and autogenous healing 54. Mihashi H, Nishiwali T (2012) Development of engineered
of cracks in concrete. ACI Mater J 96:448–455 self-healing and self-repairing concrete. J Adv Concr Technol
33. Ersan YC, Gruyaert E, Louis G, Lors C, DeBelie N, Boon N 10:170–184
(2015) Self-protected nitrate reducing culture for intrinsic repair 55. Morse JW (1983) The kinetics of calcium carbonate dissolution
of concrete cracks. Front Microbiol 6:1228 and precipitation. Rev Miner Geochem 11:227–264
34. Fowler DW (1999) Polymers in concrete: a vision for the 21st 56. Muyzer G, Stams AJ (2008) The ecology and biotechnology of
century. Cem Concr Compos 21:449–452 sulphate-reducing bacteria. Nat Rev Microbiol 6:441–454
35. Fujita Y, Redden GD, Ingram JC, Cortez MM, Ferris FG, Smith 57. Ohama Y (1998) Polymer-based admixtures. Cem Concr Compos
RW (2004) Strontium incorporation into calcite generated by bac- 20:189–212
terial ureolysis. Geochim Cosmochim Acta 68:3261–3270 58. Peckmann J, Paul J, Thiel V (1999) Bacterially mediated for-
36. Grengg C, Mittermayr F, Baldermann A, Böttcher ME, Leis A, mation of diagenetic aragonite and native sulfur in Zechstein
Koraimann G, Grunert P, Dietzel M (2015) Microbiologically carbonates (Upper Permian, Central Germany). Sediment Geol
induced concrete corrosion: a case study from a combined sewer 126:205–222
network. Cem Concr Res 77:16–25 59. Phillips AJ, Cunningham AB, Gerlach R, Hiebert R, Hwang
37. Guppy R (1988) Autogenous healing of cracks in concrete and its C, Lomans BP, Westrich J, Mantilla C, Kirksey J, Esposito R,
relevance to radwaste repositories. NSS/R—105. Nirex, United Spangler L (2016) Fracture sealing with microbially-induced cal-
Kingdom cium carbonate precipitation: a field study. Environ Sci Technol
38. Hammes F, Verstraete W (2002) Key roles of pH and calcium 50:4111–4117
metabolism in microbial carbonate precipitation. Rev Environ Sci 60. Phillips AJ, Gerlach R, Lauchnor E, Mitchell AC, Cunningham
Biotechnol 1:3–7 AB, Spangler L (2013) Engineered applications of ureolytic
39. Hearn N (1998) Self-sealing, autogenous healing and continued biomineralization: a review. Biofouling 29:715–733
hydration: what is the difference? Mater Struct 31:563–567

13
J Ind Microbiol Biotechnol (2017) 44:1511–1525 1525

61. Qian C, Chen H, Ren L, Luo M (2015) Self-healing of early age 80. Wang JY, Snoeck D, Vlierberghe SV, Verstraete W, De Belie N
cracks in cement-based materials by mineralization of carbonic (2014) Application of hydrogel encapsulated carbonate precipitat-
anhydrase microorganism. Front Microbiol 6:1225 ing bacteria for approaching a realistic self-healing in concrete.
62. Ramachandran SK, Ramakrishnan V, Bang SS (2001) Remedia- Constr Build Mater 68:110–119
tion of concrete using micro-organisms. ACI Mater J 98:3–9 81. Wang JY, Soens H, Verstraete W, De Belie N (2014) Self-healing
63. Reinhardt HW, Jooss M (2003) Permeability and self-healing of concrete by use of microencapsulated bacterial spores. Cem Concr
cracked concrete as a function of temperature and crack width. Res 56:139–152
Cem Concr Res 33:981–985 82. Wang J, Ersan YC, Boon N, De Belie N (2016) Application
64. Rodriguez-Navarro C, Rodriguez-Gallego M, Chekroun KB, of microorganisms in concrete: a promising sustainable strat-
Gonzalez-Mun˜oz MT (2003) Conservation of ornamental stone egy to improve concrete durability. Appl Microbiol Biotechnol
by Myxococcus xanthus-induced carbonate biomineralization. 100:2993–3007
Appl Env Microbiol 69:2182–2193 83. Warren LA, Maurice PA, Parmar N, Ferris FG (2001) Microbi-
65. Richardson A, Coventry K, Pasley J (2016) Bacterial crack sealing ally mediated calcium carbonate precipitation: implications for
and surface finish application to concrete. In: fourth international interpreting calcite precipitation and for solid-phase capture of
conference on sustainable construction materials and technologies inorganic contaminants. Geomicrobiol J 18:93–115

Downloaded from https://academic.oup.com/jimb/article/44/11/1511/5996420 by guest on 10 November 2021

http://www.claisse.info/Proceedings.htm 84. Wiktor V, Jonkers HM (2011) Quantification of crack-healing in
66. Sahmaran M, Keskin SB, Ozerkan G, Yaman IO (2008) Self- novel bacteria-based self-healing concrete. Cem Concr Compos
healing of mechanically-loaded self-consolidating concretes with 33:763–770
high volumes of fly ash. Cem Concr Compos 30:872–879 85. Wiktor V, Jonkers HM (2015) Field performance of bacteria-based
67. Sangadji S (2017) Can self-healing mechanism helps concrete repair system: pilot study in a parking garage. Case Stud Constr
structures sustainable? Procedia Eng 171:238–249 Mater 2:11–17
68. Seifan M, Samani AK, Berenjian A (2016) Bioconcrete: next 86. Williams SL, Kirisits MJ, Ferron RD (2016) Optimization of
generation of self-healing concrete. Appl Microbiol Biotechnol growth medium for Sporosarcina pasteurii in bio-based cement
100:2591–2602 pastes to mitigate delay in hydration kinetics. J Ind Microbiol
69. Sharma TK, Alazhari M, Heath A, Paine K, Cooper RM (2017) Biotechnol 43:567–575
Alkaliphilic Bacillus species show potential application in con- 87. Wu M, Johannesson B, Geiker M (2012) A review: self-healing
crete crack repair by virtue of rapid spore production and ger- in cementitious materials and engineered cementitious composite
mination then extracellular calcite formation. J Appl Microbiol as a self-healing material. Constr Build Mater 28:571–583
122:1233–1244 88. Xu J, Yao W (2014) Multiscale mechanical quantification of self-
70. Silva FB, Boon N, De Belie N, Verstraete W (2015) Industrial healing concrete incorporating non-ureolytic bacteria-based heal-
application of biological self-healing concrete: challenges and ing agent. Cem Concr Res 64:1–10
economical feasibility. J Commer Biotechnol 21:31–38 89. Xu J, Du Y, Jiang Z, She A (2015) Effects of calcium source on
71. Sisomphon K, Copuroglu O, Koenders EAB (2012) Self-healing biochemical properties of microbial ­CaCO3 precipitation. Front
of surface cracks in mortars with expansive additive and crystal- Microbiol 6:1366
line additive. Cem Concr Compos 34:566–574 90. Yuan YC, Rong MZ, Zhang MQ, Chen J, Yang GC, Li XM (2008)
72. Stocks-Fischer S, Galinat JK, Bang SS (1999) Microbiological Self-healing polymeric materials using epoxy/mercaptan as the
precipitation of ­CaCO3. Soil Biol Biochem 31:1563–1571 healant. Macromolecules 41:5197–5202
73. Stumm W, Morgan JJ (1981) Aquatic chemistry. John wiley, 91. Zemskov SV, Jonkers HM, Vermolen FJ (2014) A mathemati-
NewYork cal model for bacterial self-healing of cracks in concrete. J Intell
74. Tiano P, Biagiotti L, Mastromei G (1999) Bacterial bio-mediated Mater Syst Struct 25:4–12
calcite precipitation for monumental stones conservation: methods 92. Zhang JL, Wu RS, Li YM, Zhong JY, Deng X, Liu B, Han NX,
of evaluation. J Microbiol Methods 36:139–145 Xing F (2016) Screening of bacteria for self-healing of concrete
75. Tittelboom KV, De Belie N, De Muynck W, Verstraete W (2010) cracks and optimization of the microbial calcium precipitation
Use of bacteria to repair cracks in concrete. Cem Concr Res process. Appl Microbiol Biotechnol 100:6661–6670
40:157–166 93. Zhang JL, Wang CG, Wang QL, Feng JL, Pan W, Zheng XC, Liu
76. Tziviloglou E, Wiktor V, Jonkers HM, Schlangen E (2016) Bac- B, Han NX, Xing F, Deng X (2016) A binary concrete crack self-
teria-based self-healing concrete to increase liquid tightness of healing system containing oxygen-releasing tablet and bacteria
cracks. Constr Build Mater 122:118–125 and its ­Ca2+-precipitation performance. Appl Microbiol Biotech-
77. Tziviloglou E, Tittelboom KV, Palin D, Wang J, Sierra-Beltran nol 100:10295–10306
MG, Erşan YC, Mors R, Wiktor V, Jonkers HM, Schlangen E, De 94. Zhu T, Dittrich M (2016) Carbonate precipitation through micro-
Belie N (2016) Bio-based self-healing concrete: from research to bial activities in natural environment, and their potential in bio-
field application. In: Hager M, van der Zwaag S, Schubert U (eds) technology: a review. Front Bioeng Biotechnol 4:4
Self-healing materials. Advances in polymer science. Springer, 95. Zhu T, Lu X, Dittrich M (2017) Calcification on mortar by live
Cham, pp 345–385 and UV-killed biofilm-forming cyanobacterial Gloeocapsa
78. Wang JY, De Belie N, Verstraete W (2012) Diatomaceous earth as PCC73106. Constr Build Mater 146:43–53
a protective vehicle for bacteria applied for self-healing concrete. 96. Zhu T, Paulo C, Merroun ML, Dittrich M (2015) Potential appli-
J Ind Microbiol Biotechnol 39:567–577 cation of biomineralization by Synechococcus PCC8806 for con-
79. Wang J, Mignon A, Snoeck D, Wiktor V, Vliergerghe SV, Boon N, crete restoration. Ecol Eng 82:459–468
De Belie N (2015) Application of modified alginate encapsulated
carbonate producing bacteria in concrete: a promising strategy for
crack self-healing. Front Microbiol 6:1088

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