Journal of Building Engineering 70 (2023) 105930

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Journal of Building Engineering

journal homepage: www.elsevier.com/locate/jobe

Improved anti-corrosion and mechanical aspects of reinforced

cementitious composites with bio-inspired strategies
Maria Kanwal a, b, Rao Arsalan Khushnood a, *, Abdul Ghafar Wattoo c,
Muhammad Shahid d
a
NUST Institute of Civil Engineering (NICE), School of Civil and Environmental Engineering (SCEE), National University of Sciences and Technology
(NUST), H/12, Islamabad, 44000, Pakistan
b
Civil Engineering Department, Khwaja Fareed University of Engineering and Information Technology (KFUEIT), Rahim Yar Khan, 64200, Pakistan
c
Department of Physics, Khwaja Fareed University of Engineering and Information Technology (KFUEIT), Rahim Yar Khan, 64200, Pakistan
d
School of Chemical and Materials Engineering (SCME), National University of Sciences and Technology (NUST), H/12 Campus, Islamabad,
44000, Pakistan

A R T I C L E I N F O A B S T R A C T

Keywords: The device of self-healing cementitious composites is a most anticipated approach for repair of
Bacillus safensis unavoidable cracks through biological intrusions. The bio-inspired self-healing composites sys­
Sugarcane bagasse tematically heal the cracks and control the penetration of corrosive ions towards the embedded
Pyrolysis steel. However, the effect of bacterial additions on the electrochemical performance of embedded
Biochar steel holds several questions and needed to be explored further. Therefore, this research was
Self-healing
carried out to assess the effect of a calcifying bacterial strain “Bacillus safensis” and its carrier
Corrosion inhibition
media, biochar, on mechanical and transport properties along with the corrosion performance of
embedded steel in cementitious composites. The boosted strain energy storing potential with
25.5% and 38.18% enhanced flexural and compressive strength than control sample, respectively
was observed. Additionally, about 89% gain in compressive strength was observed after 28 days
of cracking at 85% of ultimate compressive strength. Moreover, a significant increase in relative
healing degree was seen during ultrasonic pulse velocity measurements of uncracked and cracked
samples. Furthermore, the healing precipitate was recognized as calcite through scanning elec­
tron microscopy and Fourier transform infrared spectroscopy. The lower sorptivity trends also
indicated the denser microstructure with minimum water uptake capability and porosity which
lead towards minimum corrosion damage in the embedded steel. Besides, the corrosion inhibition
efficiency was noted as 95.18% compared to the control sample. Thus, the proposed combination
of biotic and abiotic materials would be a good solution for recycling waste and enhancing me­
chanical and durability prospects of reinforced cementitious composites.

List of abbreviations

RCS reinforced concrete structures

RM reinforced mortar
OD optical density

* Corresponding author.
E-mail address: arsalan.khushnood@nice.nust.edu.pk (R.A. Khushnood).

https://doi.org/10.1016/j.jobe.2023.105930
Received 10 November 2022; Received in revised form 10 December 2022; Accepted 16 January 2023
Available online 18 January 2023
2352-7102/© 2023 Published by Elsevier Ltd.
M. Kanwal et al. Journal of Building Engineering 70 (2023) 105930

MICP microbially induced CaCO3 precipitation

B Bacillus
ASTM American Society of Testing Materials
GBC ground biochar
TSB tryptone soya broth
SEM scanning electron microscopy
EDS energy dispersive x-ray spectroscopy
FTIR Fourier transform infrared
UPV ultrasonic pulse velocity
MOR modulus of rupture
MOT modulus of toughness
ITZ interfacial transition zone
DD damaging degree
RHD relative healing degree

1. Introduction
The major benefit of steel as reinforcement in cementitious systems is its exceptional tensile attributes. Further, similar coefficients
of thermal expansion of steel and concrete and highly alkaline environment of concrete provides passivity to embedded steel thus act as
best composite in reinforced concrete structures (RCS) [1]. However, the composite action could be compromised under exposure of
extreme conditions. For instance, steel reinforcement de-passivate leading to active corrosion and reduction in life of RCS upon un­
dergoing severe attacks of chlorides, CO2, or stray current [2–4]. Thus, enhancing the durability of cementitious matrix and elevating
the corrosion inhibition potential of embedded steel are equally important to improve the service life of RCS.
As a crack opens in RCS, the aggressive substances including, water, sulphates, CO2, chlorides, and oxygen finds paths to penetrate
inside the cementitious matrix and influence the embedded steel [5–7]. According to a recent research, the cracks as fine as 0.1 mm
wide could provide diffusivity of chlorides inside the concrete samples subjected to salt exposures [8]. Therefore, the prompt
self-healing cementitious systems have now emerged as the sustainable strategy to tackle the cracking of structures and become the
hotspot for construction sector [9]. These approaches involve the encapsulation of crack healing agents, bacteria based strategies, use
of supplementary mineral admixtures, vascular techniques, and use of shape memory alloys [10–13]. While the bacteria based healing
approach is considered more sustainable and more efficient in healing wider cracks among other aforementioned methods [14,15].
Where the bacteria are incorporated in concrete during mixing or surface applied, the cracks get healed by the microbially induced
CaCO3 precipitation (MICP) [16–20].
Broadly, there are two approaches for supplementation of bacteria in the cementitious mix: direct addition and addition through
carrier media [21]. In the direct addition, the bacterial cells could be damaged and cause reduction in the healing efficiency of cracks
[22]. While adding via carrier media could provide shield for cells by offering resistance against (i) high shear stresses induced during
mixing, (ii) heat released during cement hydration, (iii) extreme alkaline pH, (iv) stresses due to dehydration, and (v) limited vicinity
inside the pore spaces as less as 0.1 μm [23]. Various types and sizes of bacterial carrier compounds have been tested yet whereas the
porous nano-micro sized considered superior as these particles could be coated over the surface of bacterial cells and protect them for
longer time in the extreme pH of concrete (>12) [18,24]. More conducive carrier media needs to be explored before commercializing
the bio-inspired self-healing concrete. A carrier compound endorsing bacterial preservation and contributing to elevating the dura­
bility and mechanical aspects of cementitious matrix while making it sustainable and cost efficient is still looked-for.
Biochar possesses the potential to reduce CO2 emissions and global warming prospects associated with concrete and enhances the
fracture properties [25]. In previous studies the biochar extracted from different biomasses has given boosted properties in cemen­
titious composites because of its good fracture, electromagnetic interference shielding, and thermal properties [26,27]. Use of biochar
is 94% cost efficient than using similar amount of carbon nano tubes in concrete to elevate the mechanical properties [28]. Besides, it
has also been used as bacterial carrier media in bio-inspired composites and exhibited lesser sorptivity, enhanced healing efficiency
and mechanical properties compared to the controlled formulation [29,30]. The size effect of saw-dust biochar (ground and normal
size particles) was tested on properties of mortar specimens [31]. Improvement in the early strength and reduction in water absorption
was seen through the incorporation of ground biochar (GBC) in the cementitious mix than the normal size particles [31]. Thus, the use
of fine biochar as bacterial carrier would be promising to protect bacteria during severe conditions and result in a more sustainable and
cost effective composite material to self-heal the cracks and enhance the mechanical properties.
The crack healing potential of bacteria based self-healing cementitious systems has been studied immensely with the expectation of
lesser corrosion of reinforcing bars as the MICP could block the paths of aggressive ions penetrating towards the steel. However, the
effect of healing product and bacteria on the reinforcing steel has rarely been evaluated [32,33]. Therefore, the present study is
comprised of a thorough investigation of self-healing potential and corrosion attributes of reinforced mortar (RM) specimens. A
bacterium from Bacillus specie has been selected and added through GBC (as carrier media of bacteria) to check its effectiveness on the
CaCO3 production, mechanical and transport properties along with the corrosion characteristics of embedded steel in cement mortar.

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2. Materials and methods

Ordinary Portland cement (ASTM C150 Type 1 [34]) was used as a basic and binding ingredient of mortar, with composition of
63.35% CaO, 20.35% SiO2, 4.87% Al2O3, 2.71% SO3, 2.41% MgO and 0.58% K2O. The Lawrancepur sand as the fine aggregates with
fineness modulus of 2.713 and water absorption of 2.04% as per ASTM C136 and ASTM C128 standards, respectively [35,36]. The
bacterial culture of Bacillus safensis MUGA 15 (B. safensis) strain and ground biochar (GBC) as carrier media for bacteria were the
secondary materials of mortar.
The B. safensis strain with concentration of 1.32 × 106 CFU/ml was incorporated into the cementitious mix with and without any
carrier media.

2.1. Bacterial carrier media

The bagasse, the leftover of sugarcane after sugar extraction, was transformed into an inert carbonaceous product (biochar)
through pyrolysis at 500 ◦ C. However, the detailed methodology of the pyrolysis is illustrated in our previous study [33].
Previously the porous nature of bacterial carrier compounds was preferred as pores provide house for bacteria during the harsh
mixing and hardening of cementitious samples [30]. Thus morphological investigations were conducted before and after pyrolysis
through SEM of biomass and biochar respectively. Fig. 1(a) represents the surface morphology of bagasse fibers at different resolutions.
The surface was quite porous with an estimated pore size of 900 nm. Besides, the edges of the fibers were comprised of needle like
threaded texture. However, this threaded morphology disappeared after pyrolysis. As the SEM micrographs of GBC showed
micro-meter sized irregular shaped and torturous edged sheets with some adhered nano-sized particles on the surface of sheets as
shown in Fig. 1(b). This morphology could be helping in refining the crack-paths of cementitious matrices. Moreover, the traces of
micro sized voids were seen over the surface of sheets with diameter of 0.52–1.2 μm, offering the superlative spots for shielding the
smaller B. safensis cells in harsh mixing and hydration of cement. Moreover, the larger surface area of nano particles could offer extra

Fig. 1. SEM micrographs of (a) bagasse fibers, (b) GBC and (c) EDS of GBC.

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M. Kanwal et al. Journal of Building Engineering 70 (2023) 105930

nucleation sites for the growth of hydration products during the cement hydration and could be efficient in pore-refinement and
reduced sorptivity [37]. Besides, nano-particles could be adhered over the bacterial cells during their combined sonication before
adding them in mortar mix and safeguard the cells during harsh exposures [7,15]. The composition of GBC is presented via the energy
dispersive x-ray spectroscopy (EDS) in Fig. 1(c). It is majorly composed of carbon that is an inert element and does not react with the
any compound present in matrix or hydration products of cement. The remaining minor concentrations of oxygen and calcium have
also never been reported in any negative context for concrete properties. Though, these elements could contribute to accelerating the
hydration process.
GBC was added directly or as the carrier media of B. safensis in the mortar mix after 45 min of sonication in a bath sonicator as per
the findings of our previous work [38].

2.2. Mortar mix design

The mortar samples were prepared with cement to sand contents of 1:2. Whereas, the water to binder ratio was kept 0.35 for all the
mixes as presented in Table 1. The Chemrite-303 super-plasticizer was used to have a good workable mix that was 0.45% by weight of
cement. The added water was replaced with equivalent quantity of bacterial culture, thus 4% of total water amount was replaced in
each bacterial mix [39]. Concentration of GBC was selected according to the optimum concentration of our previous work (0.2% by
cement wt.) [40]. While in the bacterial mix, GBC was acting as bacterial carrier (M4). Calcium lactate by 2% of cement weight was
used as bacterial feed. It should be noted that the calcium lactate does not compromise the mechanical attributes of cementitious
matrices, yet accelerates the MICP potential by offering plasticizing effect and extra nucleation sites effective for cement hydration [39,
41]. The mixed mortar was filled into the molds and compaction was assured through uniform tamping during filling of molds. After
24 h, samples were demolded and placed in water curing tanks at 23 ± 2 ◦ C for the defined testing ages as per ASTM C31 [42].

2.3. Mechanical properties

For flexural strength analysis plain prisms of 40 × 40 × 160 mm3 were prepared and test was conducted at a speed of 0.01 mm/min
on a servo hydraulic testing machine. However, the samples were tested at 56 days of casting.
The 50 mm3 mortar specimens were cast for compressive strength measurements and tested at 7, 14, 28 and 56 days of casting at
the loading rate of 0.25 MPa/s.
The recovery in compressive strength of damaged samples due to self-healing mechanism was evaluated by pre-cracking the cubic
specimens at 85% of respective ultimate compressive strength measured at 28 days of curing age. The samples were recurred for next
28 days to let the samples heal the induced cracks through autogenous or autonomous healing and again tested in compression. As the
bio-inspired cementitious systems recover the mechanical properties with time by precipitating minerals in induced cracks through the
metabolic actions [18]. Thus the recovery in compressive strength (RCS) was determined through Eqn. (1).
( )
F28d − Fcr
RCS (%) = 1 − × 100 (1)
F28d
Where.
F28d = 28 days compressive strength, and
Fcr = strength of cracked samples after 28 days of healing.

2.4. Ultrasonic pulse velocity (UPV) and cracks healing

The internal structure of the 40 × 40 × 160 mm3 cementitious prisms was assessed through the transmission time of ultrasonic
pulses from one extreme end of the sample to the other at a frequency of 50/60 Hz. Then velocity of transmitted pulses or UPV was
calculated using Eqn. (2). The tests were conducted at 28 days of curing before and after cracking. Then samples were placed again in
curing for 28 days and UPV was checked after 28 days of cracks initiation.
L
V= (2)
T

Where, V = ultrasonic pulse velocity (m/s).

L = length of specimen (m),
T = time of transit of pulse from one end of sample to other (sec).
As it is well known that the UPV decreases in air or water, however, travels faster in solids, which implies that UPV would be higher

Table 1
Notations and mix design of mortar samples.

Sample ID Cement (kg/m3) Sand (kg/m3) Calcium Lactate (kg/m3) Water (L/m3) Bacterial Culture (L/m3) Biochar (kg/m3)

M1 721 1442 – 252.58 – –

M2 721 1442 – 252.58 – 1.44
M3 721 1442 14.43 242.48 10.1 –
M4 721 1442 14.43 242.48 10.1 1.44

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M. Kanwal et al. Journal of Building Engineering 70 (2023) 105930

in uncracked samples and lower in cracked ones. Thus the comparative damage in the specimens in terms of damaging degree (DD) was
estimated by taking the UPV measurements of the uncracked and cracked specimens by Eqn. (3). Again upon the sealing of cracks, the
UPV could increase therefore the regain in UPV or the healing degree (HD) was also calculated by using Eqn. (4).
( )
UPV0
DD = 1 − × 100 (3)
UPVc
( )
UPVch − UPVc
HD = × 100 (4)
UPV0 − UPVc

Where, UPV0 = UPV of uncracked samples.

UPVc = UPV of cracked samples,
UPVch = UPV of cracked samples after 28 days of healing.

Since the generated cracks in each sample were uncontrolled and the extent of damage or DD could be different, therefore the direct
comparison of HD of samples was not appropriate. Thus, in order to compare the healing based on the UPV data, calculation of relative
healing degree (RHD) was deemed more accurate where the effect of DD was considered on the healing efficiency of each sample as
given in Eqn. (5).
( )
HD
RHD = × 100 (5)
1/DD
Meanwhile the visual healing analysis was also done through a digital microscope by measuring the crack widths of the healed and
unhealed cracks. The forensic evaluation of the healing product was made through SEM and Fourier transform infrared (FTIR)
spectroscopy based characterization techniques.

2.5. Rate of water absorption

The rate of water absorption or sorptivity test was conducted according to ASTM C1585 standard to assess the permeability and
densification rate of samples [43]. Specimens of 50 mm3 from each formulation were used for this test. Sorptivity was determined by
Eqn. (6) as a function of time and change in mass, when only one surface of specimen was exposed to water while others were
impenetrable.
mt
I (mm) = (6)
A×d

Where, I = the absorption (mm).

mt = the change in mass of specimen with time (g),
A = the exposed area of specimen (mm2),
d = the density of water (0.001 g/mm3).

Fig. 2. Schematic of the electrochemical test setup for reinforced mortar (RM) sample.

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M. Kanwal et al. Journal of Building Engineering 70 (2023) 105930

2.6. Electrochemical measurements

A reinforcing steel bar was embedded in addition to the previously defined materials in RM samples for electrochemical testing. The
10 mm diameter and 130 mm long deformed steel bars were used as reinforcement and were embedded at the center of the 40 × 40 ×
160 mm3 prisms as schematically shown in Fig. 2. The electric wires were connected to both ends of the steel bars before embedding to
ensure a suitable electric connection between steel bars and potentiostate. While the digital multimeter was used to check the
established connection after hardening of samples.
A three electrode based electrochemical cell was established with copper as reference, graphite as counter and RM sample as
working electrode. The electrolyte was the 3.5% NaCl solution prepared in the tap water. The Tafel polarization test was performed on
a Gamery potentiostate from − 500 mV to 500 mV at a scan rate of 0.5 mV/s. Tests were performed at six months of samples casting.
Meanwhile the RM samples were subjected to simulated marine tidal exposure by providing 10 days of wet-dry cycles through the
prepared electrolyte or the artificial marine water [8]. Tafel plots were linearly fitted to get the anodic (βa) and cathodic slopes (βc),
corrosion current densities (Icorr), and corrosion potential (Ecorr). Then the corrosion inhibition efficiency (η) with respect to the control
sample was determined through Eqn. (7) [44].
Icorr(M1) − Icorr
η= × 100 (7)
Icorr(M1)

Where.
Icorr (μA/cm2) = corrosion current densities of any RM sample and
Icorr(M1) = corrosion current densities of the control sample (M1).
The surface analysis and microstructural characteristics of the steel-concrete ITZ were done through SEM and EDS techniques.

3. Results and discussion

3.1. Load-deflection and stress-strain behavior
The flexural behavior of mortar prisms was determined as their endurance of ductility or brittleness. The load-deflection curves of
all abiotic and biotic samples with and without possession of GBC are shown in Fig. 3. The flexural toughness and strength were
observed to increase by the incorporation of GBC and B. safensis. Moreover, the curves revealed the maximum enhancement of flexural
strength in M4 sample where bacteria were added through fine GBC intrusions in the cementitious mix compared to the M1. The
modulus of rupture (MOR) and fracture toughness were calculated based on the load-deflection results. Where the MOR is the ultimate
surface stress (σ) offered by cementitious samples in three point bending test and could be evaluated by Eqn. (8) [26]. Besides, the
flexural strain (ε) was calculated by expression given in Eqn. (9).

σ or MOR = 3FmaxL/2wh2 (8)

2
ε = 6Dh/L (9)

Where, Fmax is the maximum load in the load-deflection curves, L is the effective length of prisms, w and h are the width and height of
the prism, respectively and D is the recorded maximum deflection under the applied load.
The stress-strain (σ-ε) curves were obtained from the load-deflection data which were used to estimate the modulus of toughness
(MOT) of the samples as presented in Fig. 4(a). Since the MOT also known as tensile toughness represents the amount of strain energy

Fig. 3. Load-deflection trends of all analyzed prisms.

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Fig. 4. (a) MOT, (b) flexural strength and rupture strain of analyzed samples.

stored by the material per unit volume to completely crack it. The MOT was calculated from the area under the σ-ε curves from zero
point to ultimate failure region. The control sample or M1 exhibited minimum MOT which shows its lesser strain energy keeping
capability than any other samples. The M1 showed minimum flexural strength and rupture at lesser strain compared to samples M2-M4
as shown in Fig. 4(b). However, the M2 sample being reinforced with GBC showed 31.57% enhanced strain energy storing potential
than the M1 sample. There was 13.72% enhanced flexural strength than M1 with 18.5% added rupture strain. Because the fine GBC
sheets with tortuous edges and nano-particles alter the smooth trajectory of cracks into multiple branches under the certain loading
[27]. The sharp and elongated shape of GBC micro-sheets facilitated their firm fitting in the cementitious matrix and made it to absorb
more energy at the fracture which caused high displacement of M2. Mechanical properties are typically governed by interfacial
transition zone (ITZ) that is about 10–40 μm wide in cementitious mortars [45]. From the SEM image of GBC (Fig. 1), it could be
observed that it has wide range of particle sizes. Some particles were in lesser size than the ITZ and also greater than 40 μm. Gupta et al.
[46] reported that the varying size of biochar induces strength improvement by two means, by densifying the ITZ through smaller
fraction of particles and spanning over the ITZ could be done by larger particles, thus redistribute the applied stresses and avoid brittle
failure.
The directly added microbes along with calcium lactate (M3) also contributed significantly to impart ductility and flexural strength.
That could be attributed to the MICP in pore spaces, thus offered denser matrix of M3 than the M1 and resulted in higher strength.
Besides, the bacterial mix could boost the rate of hydration as dissoluble calcium lactate content endorse lubrication in the cemen­
titious matrix [47,48]. However, the bacterial cells alone do not possess any significant effect on the hydration kinetics of fresh mix
[48]. Therefore, it could be inferred that bacterial nutrient altered the strength development pattern which caused enhancement in
strength despite of higher workability of fresh mortar in terms of endorsing fluidity [38,48]. About 17.5% enhancement in MOT and
13.7% higher flexural strength was seen by M3 than the M1 sample and the rupture strain was also closer to the M2 sample but with
lesser strength.
The M4 sample, where B. safensis were added via carbonaceous GBC sheets/particles, revealed optimum flexural attributes as

Fig. 5. Compressive strength of mortar specimens at 7, 14, 28 and 56 days of curing.

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M. Kanwal et al. Journal of Building Engineering 70 (2023) 105930

shown in Figs. 3 and 4. Because the addition of bacteria through a compatible carrier media enhanced the bacterial survival rate
compared to the direct addition in cementitious mix, thus accountable for boosted CaCO3 production and offered densified micro­
structure. Moreover, the GBC intrusions itself possess the potential of refining crack paths and enhancing strength. The flexural
strength was 25.5%, 13.67% and 22.95% more than the M1, M2, and M3 samples, respectively. Moreover, the MOT was 52.13% more
than the M1 indicating its comparatively enhanced strain keeping potential. Thus, the bacterial cells added with fine pyrolytic bagasse
biochar efficiently persuaded the tensile properties of the cementitious composites.

3.2. Compressive attributes

The compressive strength of cementitious composites is the basic property which should never be compromised with any type of
additives. Fig. 5 represents the compressive strength results of mortar cubes dosed with bacterial and non-bacterial (GBC) additions at
7, 14, 28 and 56 days of casting. The compressive strength of M1 was consistently lesser on all of the selected testing ages compared to
other bacterial and non-bacterial samples. It was 42.051 MPa at 7 days of curing age. Then kept on enhancing with the passage of time
due to the continuous hydration reactions in the cementitious composites. However, the enhancement rate of strength decreased with
the passage of time. It could be attributed to the deceleration of hydration process at later testing ages due to the major consumption of
unhydrated cement grains at the early age of hydration.
A significant increment in compressive strength was observed for M2 specimens. Due to the presence of sharp edged, varied size,
and rough surfaced GBC particles and sheets. As the GBC choked the pores due to its filler effect and contributed to elevate the strength
[49]. Moreover, the smaller sized GBC offered sufficient nucleation sites for the growth of hydration products [50]. GBC enhanced its
strength by 26.77%, 23.64%, 24.13% and 24.39% at 7, 14, 28 and 56 days of curing, respectively compared to the M1 formulation. The
significant margin of increase at 7 days curing enforces the nucleation and crack bridging action provided by the added GBC in M2
[40]. Also the fine particles behave like seeds of cement hydration, responsible for strength development at early ages due to the
production of hydration products in abundance e.g. crystallization of calcium hydroxide or calcium silicate hydrate gel takes place
earlier [46].
The M3 formulation showed 17.90%, 11.37%, 20.79% and 19.59% enhanced strength compared to the control mix (M1) at 7, 14,
28 and 56 days of curing, respectively. Thus, it could be attributed to the MICP process where the open voids were filled by the
precipitates and enhanced the load taking ability than the M1. Besides, the bacterial feed (calcium lactate) endorsed lubrication and
contributed to enhance the hydration process so it was also responsible for strengthening the M3 compared to the M1 [47,48].
However, the strength enhancement was lesser than the M2 formulation at each curing age. This could be associated to the better
refinement of pores by fine GBC intrusions than the MICP by the directly added bacteria.
While the bacterial strain added via GBC showed promising results in compression as well. The M4 offered 28.98%, 29.93%,
38.18% and 37.23% increase in compressive strength than the M1 (control) and it was 3.01%, 8.23%, 18.45% and 16.99% more than
the M2 (GBC mix), while there was 13.49%, 20.94%, 21.89% and 21.93% increase than the M3 (direct bacteria) at 7, 14, 28 and 56
days of casting, respectively. Thus, the synergistic action of B. safensis and GBC intrusions provided maximum compressive strength
than the individual incorporation of each component. Maximum strength was obtained in M4 due to the protection of bacterial cells
inside the porous GBC sheets and the nano-micro GBC particles also protected the cells by absorbing over the bacterial cell surface
during the extreme exposures of mixing and casting [51,52]. Thus, enhanced strength was achieved due to boosted deposition of
CaCO3 in voids by the bacterial metabolic actions. Previous research also supports this finding that bacteria in mortar precipitate
calcite in their surroundings which reduce the voids content and enhance the strength [16,53,54].
The gain in compressive strength as illustrated in Fig. 6, was observed in damaged specimens after 28 days of submersion in tap
water. The compressive strength was recovered as 65 ± 2.5%, 73 ± 1.75%, 78 ± 2.15% and 89 ± 2.25% for M1, M2, M3 and M4
samples, respectively. The bacteria added via GBC in cementitious matrix resulted in maximum RCS than other samples due to the most
effective healing of induced cracks in M4 mix. Besides, the added RCS is also attributed to micro reinforcement offered by fine GBC and

Fig. 6. Recovery in compressive strength after 28 days of cacks induction.

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M. Kanwal et al. Journal of Building Engineering 70 (2023) 105930

internal curing of matrix. As GBC could absorb some fraction of the mixing water, exhibit hydrophilic properties, and offer moisture to
internally cure the surrounding matrix which leads towards increased strength [55]. Due to these characteristics of GBC, it behaved
superior in M2 as well than the M1 mix. Previous studies reported that the inert GBC particles act as filler material in cementitious
matrices, thus improve the rate of hydration being micro-fillers in nature and offer place for heterogeneous nucleation [56–58].
However, in the M3, the surviving fraction of microbes healed the induced cracks through biomineralization and added in recovered
strength. As this recovered strength is more than the M2 sample, therefore it could be said that the B. safensis cells efficiently resisted
the stresses during mixing and hydration phases and also during and after induction of cracks. Whereas previously, various types of
microbes were reported killed during mixing and RCS was attained on direct induction of bacteria in cement based matrix [15,21].

3.3. Transmittance of ultrasonic pulses; pre and post healing

The UPV test was performed on uncracked and cracked samples before and after healing to estimate the densification and
improvement in quality of the internal structure. As it is well known that ultrasonic pulses transmit slowly in fractured and porous
matrices than the undamaged and sturdy ones. Likewise, it increases as the matrix hardens due to the growth and emergence of various
hydration products which densify the cementitious matrix [59,60]. Fig. 7(a) shows the recorded UPVs before and after cracking
specimens in flexural mode. It should be noted that the purpose of cracking the samples was the assessment of autonomous or
autogenous healing potential of different samples and comparative healing analysis of induced cracks. There was marginal difference
in the UPVs of different cracked and uncracked samples. However, the uncracked M4 and M2 samples showed comparatively higher
UPV as sign of denser matrix due to the nano-micro reinforcements. Because of the filler effect of GBC at the micron or sub-micron level
that accelerates the hydration process and produces well compact matrix with enhanced packing [45]. Moreover, the hydrophilic
attributes of GBC could reduce the effective w/c ratio and lessen the fraction of capillary pores due to the presence of smaller fraction of
evaporable water in the hardening phase [61,62]. Thus these factors facilitated the easy passage of pulses through the matrix and
enhanced the UPV. Out of M4 and M2 samples, the M4 was denser as evident from its UPV value due to the presence of calcium lactate
and bacterial cells in the matrix which enhanced its endurance. Similarly, in M3 sample the reason of higher UPV than M1 could be the
bacterial metabolic action to precipitate CaCO3 in pores and the comparatively more growth of hydration products in the matrix as
microbial cell itself offer extra nucleation sites for crystallization [63]. However, UPV was lesser than M4 due to the absence of inert
GBC in the M3 sample as in the uncracked samples the bacterial metabolism was limited due to the lesser availability of moisture and
oxygen in the hardened samples while the filling action of GBC is free from such dependencies.
Therefore, to compare the healing potential of different samples, they were cracked under flexural load and again UPV was checked
as presented in Fig. 7(a). Very lessen UPVs were observed after cracking with unsound differences in each sample indicating the
obvious damage and occurrence of discontinuity but with varying extent. Since the samples were cracked under uncontrolled loading
till the advent of visible cracks therefore the damage level was different in each sample. Thus to make the results valid for comparison,
the DD and respective RHD was calculated as shown in Fig. 7(b). The damaging degree was not depending on the quality of matrix
therefore maximum damage was observed in M4, then in M2, M1 and M3 respectively. However, the RHD was an important outcome
representing the self-healing potential of induced cracks with respect to each other.
Maximum RHD as indicator of maximum healing of cracks was observed in M4 sample. It endorses the better preservation of
B. safensis by porous GBC sheets/particles resulting maximum amount of CaCO3 production to heal the wider cracks by bacterial
metabolic activities. Additionally the autogenous healing was also contributing by hydration of unhydrated cement grains available at
cracks faces or in exposed-voids by reacting with water during the submersion-curing after induction of cracks. Furthermore, the
carbonation of calcium hydroxide to produce CaCO3 could also add in the healing of cracks due to the availability of CO2 as explained
by Eqn. (10).

Fig. 7. Characteristics of internal structure of mortar by (a) UPV of uncracked and cracked samples (b) induced DD and RHD by mineralization.

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M. Kanwal et al. Journal of Building Engineering 70 (2023) 105930

CO2 + Ca(OH)2 → CaCO3 + H2O (10)

However, the CO2 could be either coming from outer environment or inner matrix, as bacteria produce CO2 during their metabolic
action while precipitating CaCO3 by consuming the calcium source as shown by reaction in Eqn. (11).
̅̅̅̅̅→
CaC6 H10 O6 + 6O2 Bacteria CaCO3 + 5CO2 + 5H2 O (11)
Moreover, there could be minor contribution from the additional growth of hydration products over large surface area of fine GBC
[64,65]. Thus integration of all these factors from autonomous and autogenous healing processes made the M4 as most promising
sample with boosted healing potential of cracks.
The M3 sample showed only 13.46% lesser healing potency than the M4, however it was 47.57% and 44.04% more than the M1 and
M2 samples, respectively. Thus the RHD confirmed the result of the strength regain of cracked samples, that the direct induction of
B. safensis was also somehow more effective than the control and biochar added samples. While the M1 and M2 represented quite
similar RHD with marginal difference due to the limited sources of added healing and autogenous healing was lesser effective than the
autonomous or bioinspired healing of cracks. Some of the visually healed cracks by all samples are presented in Fig. 8. The M1 and M2
samples did not heal the visual cracks but the enhanced UPVs represented that there could be microlevel healing which contributed to
enhance the density and added into RHD. The directly added microbes in M3 showed substantial RHD but lesser than M4 and the
cracks healing was also nonhomogeneous as seen from Fig. 8(c). This could be attributed to the unavailability of microbial cells along
the full length of cracks due to their lesser survival in direct incorporation. Thus the available microbes were only able to precipitate in
their limited surrounding area which resulted in the cracks healing in varying length of patches instead of homogeneous healing
throughout the length of crack. Consequently, the complete closure of cracks took longer time than M4 thus reduced the healing rate.
In M3, maximum of 1.668 mm width of cracks were completely sealed by bacterial metabolically precipitate and the 1.964 mm crack
was also partialy healed with minor unhealed portion of about 187 μm. However, it was still expected that the remaining portion
would be healed with the exceedance of healing period to more than 28 days. The B. safensis added via GBC (M4) precipitated
abundance of CaCO3 and completely healed a varying width of crack from 1.823 mm to 2.532 mm as shown in Fig. 8(d).
The bacterial precipitated product was carefully scraped from cracks and analyzed through SEM as shown in Fig. 9. It was noted
that the most of the crystals were agglomerated with each other and possessed different type of microstructural characteristics
including varing size and shape. The shapes of the precipitates were found varying from small ovel crystals to large glubular or
spherical which were somehow amorphous in nature with uneven surface. These varied size and clustered grains contributed to
compactly block the cracks and improved the density of matrix.
Besides, clear bacterial imprints were seen in the precipitates which were nearly equivalent to the size of the B. safensis cells [66,
67]. Since the size of the cavities ranged between 0.47 μm and 0.81 μm. The possible reason for the escape of bacterium from such sites
or development of such holes could be the unavailability of proper nutrients, air and water during the crystalization of CaCO3 around
the cells. Thus living environment became really harsh for B. safensis and they might escaped from the crystals with the help of their

Fig. 8. Cracks healing by different formulations (a) M1, (b) M2, (c) M3, and (d) M4.

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M. Kanwal et al. Journal of Building Engineering 70 (2023) 105930

Fig. 9. (a) SEM image of product healing the cracks and (b) enlarged view of bacterial imprints in precipitates.

flagellum and left some imprints in the precipitates [68]. Previous few studies concerning growth evolution of CaCO3 also support this
phenomenon [69].
The FTIR spectroscopy being helpful in identifying the materials composition was used to confirm the bacterial precipitate as
CaCO3. The FTIR spectra is shown in Fig. 10 against wavenumber in 1/cm (cm− 1) units and transmittance in arbitrary units (a.u.). In
the spectrum the adsorption band around 3400 cm− 1 was the fingerprint of absorbed water molecules in the precipitate. Between 2250
cm− 1 and 3200 cm− 1 were the indication of the vibration of C–H stretching bonds. However, the adsorption bands of carbonates could
be divided into four segments i.e. (i) symmetric stretch of CO2−
3 around 1080 cm
− 1
that is marked as v1 on spectrum, (ii) out of plane
bending around 870 cm− 1 which is v2, (iii) asymetric stretching around 1400 cm− 1 as v3 and (iv) in-plane bending at about 700 cm− 1
as v4. Besides the bands around 700, 875 and 1425 cm− 1 also represent the out of plane and asymmetric stretching vibrations of O–C–O
bonds respectively. And the sharp absorption bands of v2 and v4 confirms the formation of calcite by bacterial activity and it was
according to the previously reported IR spectrums of calcite [70,71].

3.4. Sorptivity behavior; pre and post healing

The sorptivity results of uncracked (M1-M4) and cracked (M1’-M4’) samples are presented in Fig. 11. Initially abrupt rise in water
absorption was observed but the secondary rate was lesser and became negligible at later days in each undamaged sample. It could be
according to the saturation capacity of each sample.
Overall, in the biochar based undamaged samples (M4 and M2) minimum sorptivity was observed due to the denser and refined
matrix with lesser pores in size and number as result of additional crystallization of minerals in the voids as shown in Fig. 12(a). The
hydrophilic GBC could internally cure the matrix due to having higher water retaining capability and provide moisture during
hardening of sample thus adds in hydration process to produce products in surrounding pores. The porosity or number and size of pores
in the cementitious matrices mostly dominate in the water absorption capacity. Initially finer pores hold the water through capillary
action and influence the sorptivity of earlier days [46]. Upon saturation of fine pores, the larger capillary pores then air voids adsorb
water and cause enhancement of secondary sorptivity. Whereas the specific proportion of pores comprising capillary and dead ended
pores, contributing to retaining the water, was previously termed as effective active-porosity [72]. Besides, the high water absorption

Fig. 10. FTIR of bacterial precipitate in cracks.

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M. Kanwal et al. Journal of Building Engineering 70 (2023) 105930

Fig. 11. Sorptivity trends of analyzed specimens representing the porosity of cracked and uncracked specimens.

Fig. 12. SEM micrographs of (a) hydration products developing in voids and (b) autogenous healing or carbonation of calcium hydroxide in micro voids and cracks.

of biochar may reduce the amount of free water in fresh mortar resulting in reduced local w/c ration in the hardened matrix. Then may
induce lesser capillary pores due to the presence of lesser amount of evaporable water [73].
Besides, M4 was the most promising in retention of lesser amount of water as bacteria added through GBC got activated upon the
ingress of water and started their metabolic activity resulting in crystallization of CaCO3 in the voids. Thus made the microstructure
denser and hindered the penetration and retention of water inside the matrix as evident from sorptivity trends in Fig. 11. The M3
sample also exhibited quite denser microstructure in terms of showing high resistance for sorptivity compared to the M1 but this

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M. Kanwal et al. Journal of Building Engineering 70 (2023) 105930

resistance was lower than the M4 sample. Thus due to the possibility of bacterial damage in direct incorporation and absence of fine
reinforcement like GBC in M2 made it comparatively less effective than M4. However, the biomineralization occurred by survived
bacteria and nucleation effects of added micro sized bacterial cells with calcium lactate facilitated the production of CaCO3 and growth
of hydration products [30].
It was further observed that the M4 sample showed reduced initial and secondary sorptivity in both cracked (M4’) and uncracked
conditions. It means that the carrier media of bacteria protected the cells and remained available for mineralizing during cracked and
uncracked conditions. Thus efficiently maintained the water tightness of matrix in pre and post healing phases. However in the cracked
sample a throughout increase in absorption was seen. This might be attributed to the continuous formation of healing products in the
induced cracks in addition to the absorption of water and resulted in mass gain of samples thus directly affected the sorptivity (I)
despite of any limitation of saturation capacity of samples [43]. While this behavior was not that prevalent in cracked control sample
(M1’). The M2’ and M3’ also somehow followed the behavior of M4’ in continuously gaining weight. Still, M2’ was behaving uniquely
than any other samples as there was uneven trend of cracked sample. It could be due to large water absorption potential of biochar
during the upwards seepage of water in samples, therefore they exhibited gain in sorptivity at the interaction of any GBC intrusions
with the water. Moreover, autogenous healing could also impart a bit gain in weight and affect the sorptivity trends. Micro-cracks and
fissures could be healed in cracked samples when water reacts with unhydrated grains of cement available at cracks faces and edges of
voids. Furthermore, the carbonation of Ca(OH)2 may also be involved in the autogenous healing of cracked samples. Because during
the penetration of water inside the cementitious matrix it may bring CO2 from the external environment with it which could laterally
react with Ca(OH)2 to produce CaCO3 [74,75]. Similar behavior was detected by the SEM micrograph presented in Fig. 12(b). The
curing of cracked samples resulted in the formation of spongy-fibrous CSH gel in the available spaces and crystallization of Ca(OH)2
occurred [13]. The morphology of Ca(OH)2 in Fig. 12(b) was quite different than its plate like structure reported previously and in
Fig. 12(a) [76,77].
Therefore, it could be thought that the transformation of Ca(OH)2 to CaCO3 occurred due to carbonation. Since this morphology of
CaCO3 was different than the biological precipitated product shown in Fig. 9, it was confirmed that this product was produced
autogenously instead of autonomous healing. Moreover, the CaCO3 produced by microbial activity was more in amount, somehow
amorphous and spherical in shape. While the carbonation of calcium hydroxide formed lesser in quantity, crystalline in nature and
well-arranged plates or somewhat rhombohedral shape of calcite. Thus MICP is a more proficient way to heal wider cracks without
compromising the alkalinity of cementitious systems, unlike carbonation, where pH of cement mortar drops with the transformation of
calcium hydroxide to calcium carbonate.

3.5. Electrochemical attributes

Normally reinforcing steel bars are protected by a 5 nm–13 nm thick passive film of iron oxyhydroxide maintained at a pH of more
than 12 of the capillary interstitial solution [78]. Unfortunately, the contact of some hostile ions especially chloride negatively affect
the passivity leading to unstable electrochemical aspects and corrosion of embedded steel. The structures located in marine envi­
ronment are more prone to chlorides and experience corrosion induced destructions [79]. Therefore, the effect of chlorides on the
electrochemical behavior of bio-inspired reinforced cementitious samples was crucial to find. The electrochemical or Tafel polarization
results of all RM samples are shown in Fig. 13. The current density (I) along the x-axis is the electric current density generated during
the transferring of electrons from electronegative region to the electropositive region with the acceleration of corrosion on the working
electrode. In short, the higher the I value represents the higher level of corrosion in the working electrode. Under natural conditions,
corrosion current density (Icorr) varies from 0.1 μA/cm2 to 10 μA/cm2 and under severe conditions it may reach to 100 μA/cm2 [80].
While the severe conditions might be the cracking in cementitious matrix, direct contact of aggressive ions to steel bars or small
damage in the steel coatings etc. A smaller anodic area is more dangerous than the larger one because it permits larger anodic current

Fig. 13. Tafel curves of analyzed samples at 180 days of wet-dry exposure of chlorides.

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M. Kanwal et al. Journal of Building Engineering 70 (2023) 105930

to flow and accelerates the corrosion.

Besides, the corrosion potential (Ecorr) on the y-axis represents the polarization of the working electrode with respect to the
reference electrode. The larger the difference between the potentials of reference and working electrodes represents the higher the
level of corrosion and vice versa. The ASTM C876 defines the Ecorr ranges for the different levels of the possibility of corrosion in
reinforced cementitious samples [81]. If the Ecorr is more negative than − 350 mV, then it shows corrosion is happening in steel with
90% probability. If it is between − 200 mV and − 350 mV, then corrosion is uncertain. While for lesser negative potential than − 200 mV
there is more than 90% probability of no corrosion in reinforcing steel. But the estimation of corrosion attributes of steel just on the
basis of potential have not been much encouraged by some of the studies as Ecorr shows the qualitative estimation of corrosion while
Icorr is a quantitative representer [32,82]. Ecorr and Icorr data extracted from Fig. 13 is given in Table 2. The M1 sample showed most
negative Ecorr than other samples that was about − 260 mV indicating the uncertaininity of corrosion while Icorr was 21.82 μm/cm2 that
is far away from the natural corrosion value thus represented the accelerated corrosion of embedded steel as per the previous discussed
two criteria. M2 sample being supplemented with nano-intrusions effectively hindered the penetration of chlorides inside the matrix
and reduced the Ecorr to some extent and Icorr was also exceptionally reduced and remained within the natural corrosion range. The
sorptivity results also support this finding that M2 possessed minimum sorptivity after M4 sample, thus the penetration rate of
chlorides was slower. Similar to the M1 and M2 samples the corrosion was uncertain in the M3 sample, as Ecorr was lying in the range of
− 200 mV to − 350 mV [81]. However the Icorr was in the natural or free corrosion range thus it was corroding slowly due to the
resistance offered by directly inducted microbes along with the calcium lactate and their precipitated products. Besides, this enhanced
potential to resist corrosion than the M1 could be due to the presence of excessive calcium lactate around the surrounding of embedded
steel. Because it could offer sufficient amount of calcium ions for the production of Ca(OH)2 gel having high pH thus leads to reduction
of corrosion possibility in steel bar [33]. Moreover, the absorption of carboxylic groups from the added bacterial nutrients could also
prevent the contact of chlorides with steel [83]. Therefore, a detailed study on the effect and consequences of calcium lactate on steel
bars in alkaline environments is recommended.
Additionally the corrosion could be protected by consumption of available oxygen by bacteria during their metabolic process and
yielding of healing products. Though, the presence of oxygen is needed to initiate corrosion as cathodic reaction could not proceed
without its availability and corrosion initiation is equally controlled by cathodic and anodic reactions as shown by Eqn. (12) to Eqn.
(14).

Fe → Fe2+ +2e− (anodic reaction) (12)

½ O2 + H2O +2e → 2OH (cathodic reaction)

− −
(13)
2+
Fe + OH → Fe(OH)2 (formation of corrosion products)
−
(14)

Most promising results were seen in the M4 sample, according to the above discussed criteria of interpretation of electrochemical
test results this sample was safer in corrosion. The Ecorr was − 5.4 mV that is quite lesser than the − 200 mV, and Icorr was also very close
to the lower value of natural corrosion range. Thus the M4 sample remained corrosion impeded during the exposure period of chlorides
for 180 days due to the synergetic protections provided by various combined sources like GBC, bacteria, and calcium lactate. The M4
mix was 95.18% more efficient than the M1 mix in inhibiting the corrosion. Whereas the mechanism of corrosion protection in M4
sample was the combined effect of all the factors acting individually in M1, M2 and M3 samples.
Additionally, a whitish layer was noticed on the surface of reinforced M4 and M3 samples submerged in the 3.5% solution of NaCl.
It was thought that it could be the deposits of salt just over the surface due to the resistance offered by denser matrix to penetrate them
inside or could be the bacterial precipitate as observed by a previous study on bio-self-healing composites during water absorption test
[30]. Therefore, it was investigated through SEM and EDS analysis as presented in Fig. 14(a–c). The results showed that there were
huge number of deposits of spherical to round shaped CaCO3 mineral with smaller amount of NaCl which were well spread on the
sample’s surface. The shape of the deposited CaCO3 was well matching with the microbially precipitated CaCO3 in the cracks of
bacterial samples as shown by Fig. 9. However the observed shape was different than the shape presented in Fig. 12(b) due to the
autogenous healing. Thus the production of CaCO3 by bacteria lying near the surface of RM samples was confirmed because of the
contact of water with the near available cells. The bio precipitate contributed to filling the open voids and lowered the penetration of
chlorides and protected the samples from corrosion.
For the evaluation of microstructural characterizations of ITZ of the modified samples the SEM and EDS analysis was done as
presented in Fig. 15(a and b). The M4 sample where B. safensis were added through GBC, showed minor content of iron at the ITZ of
steel and mortar compared to the M1 sample which could be attributed to the lesser movement of ions or lesser corrosion of the
embedded steel surface. Because the any penetrated oxygen during the analysis period was consumed by bacteria to biomineralize.
Moreover, the calcium weightage was also significantly higher than the M1 indicating the presence of calcium lactate or CaCO3
precipitated by the bacterial metabolism at the open spaces of mortar in addition to the CSH and Ca(OH)2. Besides, the SEM image

Table 2
Electrochemical properties of analyzed RM specimens.

Electrochemical parameters M1 M2 M3 M4

Icorr (μA/cm2) 21.82 1.08 2.21 1.05

Ecorr (mV) − 260 − 225 − 215 − 5.4

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M. Kanwal et al. Journal of Building Engineering 70 (2023) 105930

Fig. 14. (a) M4 surface with whitish layer (b) SEM micrograph of precipitate over surface (c) EDS spectra representing the composition of surfacial layer.

presented relatively compact and improved microstructure of matrix at ITZ. Thus the M4 bacterial formulation efficiently densified the
ITZ and inhibited the penetration of chlorides to reach the embedded steel and consequently inhibited corrosion.

3.6. Cost analysis

For the cost analysis of bio-inspired mortar the material’s prices in USD were confirmed from the international suppliers and used to
calculate the cost for a cubic meter of mortar as shown in Table 3. Whereas the unit price for B. safensis was picked from ATCC website
[84]. It is pertinent to mention that the biological self-healing mortar possess significantly higher cost than the conventional mortar for
a cubic meter. While the bio-inspired concrete is more efficient for structures exposed to aggressive environments where frequent
repair and maintenance is required thus the long term costs or repair costs could be eliminated by using this cocnrete. For example, the
structures located in chlorides or marine environment and alongside the water reservoirs are prone to corrosion thus require costly and
repeated repairs. So by using bio-inspired self-healing concrete the manual inspection and repair costs could be reduced. In the current
study the bio-inspired mortar added about 16.85% cost to the conventional mortar, while the cost is significantly less than a calcium
nitrite based mortar which is a commercially available corrosion inhibitor. Whereas an optimum amount of calcium nitrite as corrosion
inhibitor of reinforced concrete is 2–4% by weight of cement [85] therefore a middle amout, 3% by cement weight, was considered for
cost analysis which was estimated according to the Sigma Aldrich rates [86]. Thus by using the bio-inspired concrete about 11.3% cost
could be avoided than using the local commericial corrosion inhibitors. It should be noted that the cost of bio-inspired concrete could
be significantly reduced by using locally available materials (e.g., bacterial nutrients and bacteria) as done in the current research.
Where the B. safensis were isolated from alkaline soils and locally available calcium lactate was used.

4. Conclusion
In this research, the B. safensis bacteria were added in the cementitious composites through the fine bagasse biochar as carrier
media. The effects of bacteria and biochar on the mechanical, sorptivity, ultrasonic pulse velocity, healing and corrosion performance
of plain and reinforced mortar have been investigated, by adding them individually and collectively into the mix. So following con­
clusions are drawn.
(a) It is found that adding B. safensis with sugarcane-bagasse biochar improves the compressive and flexural properties of
cementitious samples. That implies the synergistic effect of biochar and bacteria in densification of mortar by well-preservation
of microbes leading to enhancement in bacterial metabolically precipitated calcite in voids.

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M. Kanwal et al. Journal of Building Engineering 70 (2023) 105930

Fig. 15. Microstructural and elemental analysis of steel-mortar ITZ by SEM and EDS analysis.

Table 3
Cost comparison of bio-inspired mortar samples with a commericial inhibitor.

Material Unit Price (USD) M1 (USD/m3) M2 (USD/m3) M3 (USD/m3) M4 (USD/m3) Commercial Inhibitor (USD/m3)

Cement 0.089 64.17 64.17 64.17 64.17 64.17

Sand 0.0089 12.83 12.83 12.83 12.83 12.83
GBC 10 – 14.4 – 14.4 –
B. safensis 590 – – 98 98 –
Calcium Lactate 16 – – 230.88 230.88 –
Bacterial growth media or TSB 120 – – 36.6 36.6 –
Calcium Nitrite 183 – – – – 3958.29
Total 77 91.4 442.48 456.88 4035.29

(b) The bacterial samples added via carrier media show high compressive strength recovery of cracked samples compared to the
directly incorporated bacteria and control samples by completely filling the induced cracks through CaCO3.
(c) Higher ultrasonic pulse velocity of undamaged sample and higher relative damaging degree by M4 sample is due to the
improved microstructural quality and complete healing of cracks by MICP.
(d) There is a clear difference in the bacterially produced calcite and the calcite produced through carbonation of calcium hy­
droxide. The calcite produced through bacteria is more in amount, less crystalline and spherical in shape while carbonation
leads to produce lesser amount, more crystalline and plates to rhombohedral shape. Thus MICP is efficient in healing wider
cracks than carbonation while maintaining the alkalinity.
(e) The co-added bacteria and biochar effectively reduces the water absorption rate of uncracked specimens due to the well-
compacted matrix by bacterial calcite deposits in pores and surface of samples. Besides, provides effective recovery in sorp­
tivity of cracked samples by healing the induced cracks and densifying the matrix.
(f) The M4 samples gave minimum corrosion in embedded steel due to the enhanced passivity inside the bio-inspired matrix
provided by calcium lactate, bacterial calcite deposits at surface, voids and ITZ in addition to the well-refined matrix offered by
biochar intrusions. Thus inhibits the penetration of chlorides towards steel and protects the steel bars from corrosion.

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M. Kanwal et al. Journal of Building Engineering 70 (2023) 105930

However, it is further recommended to explore the comparatively cheaper nutrients of bacteria for their invitro growth and survival
in concrete so that the cost difference between conventional and bio-inspired concrete could be reduced. Moreover the freeze-thaw
characteristics along with the high temperature tolerance of bacteria should also be checked to optimize the use of bio-inspired
concrete in extreme environmental conditions.

Author’s contribution statement

Maria Kanwal: Conceptualization, Investigation, Writing. Rao Arsalan Khushnood: Conceptualization, Supervision, Review &
Editing and Resources. Abdul Ghafar Wattoo: Resources, Writing, and Editing. Muhammad Shahid: Supervision, Resources.

Declaration of competing interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to
influence the work reported in this paper.

Data availability

No data was used for the research described in the article.

Acknowledgements
The research presented in this paper was sponsored by Higher Education Commission (HEC), Islamabad, Pakistan under research
grant# 10232/Federal/NRPU/R&D/HEC/2017.

References
[1] J. Shi, X. Guan, J. Ming, X. Zhou, Improved corrosion resistance of reinforcing steel in mortars containing red mud after long-term exposure to aggressive
environments, Cem. Concr. Compos. 130 (2022), 104522, https://doi.org/10.1016/J.CEMCONCOMP.2022.104522.
[2] M. Stefanoni, U. Angst, B. Elsener, Corrosion rate of carbon steel in carbonated concrete – a critical review, Cement Concr. Res. 103 (2018) 35–48, https://doi.
org/10.1016/J.CEMCONRES.2017.10.007.
[3] L. Bertolini, M. Carsana, P. Pedeferri, Corrosion behaviour of steel in concrete in the presence of stray current, Corrosion Sci. 49 (2007) 1056–1068, https://doi.
org/10.1016/j.corsci.2006.05.048.
[4] L. Chen, R.K.L. Su, Corrosion rate measurement by using polarization resistance method for microcell and macrocell corrosion: theoretical analysis and
experimental work with simulated concrete pore solution, Construct. Build. Mater. 267 (2021), 121003, https://doi.org/10.1016/j.conbuildmat.2020.121003.
[5] L. Jiang, D. Niu, Study of deterioration of concrete exposed to different types of sulfate solutions under drying-wetting cycles, Construct. Build. Mater. 117
(2016) 88–98, https://doi.org/10.1016/J.CONBUILDMAT.2016.04.094.
[6] Y. Chen, J. Gao, L. Tang, X. Li, Resistance of concrete against combined attack of chloride and sulfate under drying–wetting cycles, Construct. Build. Mater. 106
(2016) 650–658, https://doi.org/10.1016/J.CONBUILDMAT.2015.12.151.
[7] W. Khaliq, M.B. Ehsan, Crack healing in concrete using various bio influenced self-healing techniques, Construct. Build. Mater. 102 (2016) 349–357, https://doi.
org/10.1016/j.conbuildmat.2015.11.006.
[8] M.B. Ehsan Khan, L. Shen, D. Dias-da-Costa, Crack healing performance of bacteria-based mortar under sustained tensile loading in marine environment, Cem.
Concr. Compos. 120 (2021), 104055, https://doi.org/10.1016/J.CEMCONCOMP.2021.104055.
[9] M. Park, S. Park, J.Y. Yoo, Y. Kim, K.M. Lee, D.Y. Hwang, H.J. Son, Enzyme-mediated biocalcification by a novel alkaliphilic Bacillus psychrodurans LC40 and its
eco-friendly application as a biosealant for crack healing, Sci. Total Environ. 802 (2022), 149841, https://doi.org/10.1016/J.SCITOTENV.2021.149841.
[10] O.R. Teall, Crack Closure and Enhanced Autogenous Healing of Structural Concrete Using Shape Memory Polymers, Cardiff University, 2016. https://orca.
cardiff.ac.uk/id/eprint/100250.
[11] L. Restuccia, A. Reggio, G.A. Ferro, J.-M. Tulliani, New self-healing techniques for cement-based materials, Procedia Struct. Integr. 3 (2017) 253–260, https://
doi.org/10.1016/J.PROSTR.2017.04.016.
[12] E. Schlangen, H. Jonkers, S. Qian, A. Garcia, Recent advances on self healing of concrete,Fract, Mech. Concr. Concret. 8 (2010) 1–08. https://framcos.org/
FraMCoS-7/02-10.pdf.
[13] K. Van Tittelboom, N. De Belie, Self-healing in Cementitious Materials-A Review, 2013, https://doi.org/10.3390/ma6062182.
[14] S. Marín, O. Cabestrero, C. Demergasso, S. Olivares, V. Zetola, M. Vera, An indigenous bacterium with enhanced performance of microbially-induced Ca-
carbonate biomineralization under extreme alkaline conditions for concrete and soil-improvement industries, Acta Biomater. 120 (2021) 304–317, https://doi.
org/10.1016/J.ACTBIO.2020.11.016.
[15] N. Shaheen, R.A. Khushnood, W. Khaliq, H. Murtaza, R. Iqbal, M.H. Khan, Synthesis and characterization of bio-immobilized nano/micro inert and reactive
additives for feasibility investigation in self-healing concrete, Construct. Build. Mater. 226 (2019) 492–506, https://doi.org/10.1016/j.
conbuildmat.2019.07.202.
[16] N. Karimi, D. Mostofinejad, Bacillus subtilis bacteria used in fiber reinforced concrete and their effects on concrete penetrability, Construct. Build. Mater. 230
(2020), 117051, https://doi.org/10.1016/j.conbuildmat.2019.117051.
[17] Y. Su, T. Zheng, C. Qian, Application potential of Bacillus megaterium encapsulated by low alkaline sulphoaluminate cement in self-healing concrete, Construct.
Build. Mater. J. 273 (2021), 121740, https://doi.org/10.1016/j.conbuildmat.2020.121740.
[18] R.A. Khushnood, S. ud din, N. Shaheen, S. Ahmad, F. Zarrar, Bio-inspired self-healing cementitious mortar using Bacillus subtilis immobilized on nano-/micro-
additives, J. Intell. Mater. Syst. Struct. 30 (2019) 3–15, https://doi.org/10.1177/1045389X18806401.
[19] C. Neeladharan, Application of Bacillus subtilis bacteria for improving properties and healing of cracks in concrete, Int. J. Adv. Res. Trends Eng. Technol. 5
(2018) 1–7, https://doi.org/10.20247/IJARTET.2018.05S05030023.
[20] W. De Muynck, D. Debrouwer, N. De Belie, W. Verstraete, Bacterial carbonate precipitation improves the durability of cementitious materials, Cement Concr.
Res. 38 (2008) 1005–1014, https://doi.org/10.1016/j.cemconres.2008.03.005.
[21] T.H. Nguyen, E. Ghorbel, H. Fares, A. Cousture, Bacterial self-healing of concrete and durability assessment, Cem. Concr. Compos. 104 (2019), 103340, https://
doi.org/10.1016/J.CEMCONCOMP.2019.103340.
[22] N. Shaheen, R.A. Khushnood, M.A. Musarat, W.S. Alaloul, Self-healing nano-concrete for futuristic infrastructures: a review, Arabian J. Sci. Eng. 47 (2022)
5365–5375, https://doi.org/10.1007/S13369-022-06562-6/TABLES/1.
[23] J.Y. Wang, N. De Belie, W. Verstraete, Diatomaceous earth as a protective vehicle for bacteria applied for self-healing concrete, J. Ind. Microbiol. Biotechnol. 39
(2012) 567–577, https://doi.org/10.1007/S10295-011-1037-1.

17
M. Kanwal et al. Journal of Building Engineering 70 (2023) 105930

[24] N. Shaheen, R.A. Khushnood, S. Ud Din, A. Khalid, Influence of bio-immobilized lime stone powder on self-healing behaviour of cementitious composites, IOP
Conf. Ser. Mater. Sci. Eng. 431 (2018), https://doi.org/10.1088/1757-899X/431/6/062002.
[25] A. Khalid, R.A. Khushnood, S. Ali Memon, Pyrolysis as an alternate to open burning of crop residue and scrap tires: greenhouse emissions assessment and
mechanical performance investigation in concrete, J. Clean. Prod. 365 (2022), 132688, https://doi.org/10.1016/j.jclepro.2022.132688.
[26] A. Khalid, R.A. Khushnood, A. Mahmood, Impact of pyrolytic carbonaceous nano inerts addition on fracture and electromagnetic interference shielding
characteristics of cementitious composites, Theor. Appl. Fract. Mech. 103 (2019), 102320, https://doi.org/10.1016/j.tafmec.2019.102320.
[27] R.A. Khushnood, S. Ahmad, G.A. Ferro, L. Restuccia, J.M. Tulliani, P. Jagdale, Modified fracture properties of cement composites with nano/micro carbonized
bagasse fibers, Frat. Ed. Integrità Strutt. 9 (2015) 534–542, https://doi.org/10.3221/IGF-ESIS.34.59.
[28] A. Mahmood, R.A. Khushnood, M. Zeeshan, Pyrolytic carbonaceous reinforcements for enhanced electromagnetic and fracture response of cementitious
composites, J. Clean. Prod. 248 (2020), 119288, https://doi.org/10.1016/J.JCLEPRO.2019.119288.
[29] H.W. Kua, S. Gupta, A.N. Aday, W.V. Srubar, Biochar-immobilized bacteria and superabsorbent polymers enable self-healing of fiber-reinforced concrete after
multiple damage cycles, Cem. Concr. Compos. 100 (2019) 35–52, https://doi.org/10.1016/J.CEMCONCOMP.2019.03.017.
[30] S. Gupta, H.W. Kua, S.D. Pang, Healing cement mortar by immobilization of bacteria in biochar: an integrated approach of self-healing and carbon
sequestration, Cem. Concr. Compos. 86 (2018) 238–254, https://doi.org/10.1016/J.CEMCONCOMP.2017.11.015.
[31] S. Gupta, H.W. Kua, Carbonaceous micro-filler for cement: effect of particle size and dosage of biochar on fresh and hardened properties of cement mortar, Sci.
Total Environ. 662 (2019) 952–962, https://doi.org/10.1016/J.SCITOTENV.2019.01.269.
[32] J. Xu, Y. Tang, X. Wang, Z. Wang, W. Yao, Application of ureolysis-based microbial CaCO3 precipitation in self-healing of concrete and inhibition of
reinforcement corrosion, Construct. Build. Mater. 265 (2020), 120364, https://doi.org/10.1016/j.conbuildmat.2020.120364.
[33] M. Kanwal, R.A. Khushnood, M. Shahid, A.G. Wattoo, An integrated and eco-friendly approach for corrosion inhibition and microstructural densification of
reinforced concrete by immobilizing Bacillus subtilis in pyrolytic sugarcane-bagasse, J. Clean. Prod. 355 (2022), 131785, https://doi.org/10.1016/j.
jclepro.2022.131785.
[34] American Society of Testing Materials, ASTM C150/C150M - 18 Standard Specifications for Portland Cement, 2012, https://doi.org/10.1520/C0150.
[35] American Society of Testing Materials, Standard Test Method for Sieve Analysis of Fine and Coarse Aggregates (ASTM C136), Annu. B. ASTM Stand, 2006,
pp. 3–7, https://doi.org/10.1520/C0136.
[36] American Society of Testing Materials,, Standard Test Method for Relative Density (Specific Gravity) and Absorption of Fine Aggregates, ASTM C128), 2015,
https://doi.org/10.1520/C0128-15.2.
[37] A. Mahmood, R.A. Khushnood, M. Zeeshan, Pyrolytic carbonaceous reinforcements for enhanced electromagnetic and fracture response of cementitious
composites, J. Clean. Prod. 248 (2020), 119288, https://doi.org/10.1016/j.jclepro.2019.119288.
[38] M. Kanwal, R.A. Khushnood, W. Khaliq, A.G. Wattoo, T. Shahid, Synthesis of pyrolytic carbonized bagasse to immobilize Bacillus subtilis; application in healing
micro-cracks and fracture properties of concrete, Cem. Concr. Compos. 126 (2022), 104334, https://doi.org/10.1016/j.cemconcomp.2021.104334.
[39] N. Shaheen, A. Jalil, F. Adnan, R.A. Khushnood, Isolation of Alkaliphilic Calcifying Bacteria and Their Feasibility for Enhanced CaCO 3 Precipitation in Bio-
Based Cementitious Composites, 2021, pp. 1–16, https://doi.org/10.1111/1751-7915.13752.
[40] R.A. Khushnood, S. Ahmad, G.A. Ferro, L. Restuccia, J.M. Tulliani, P. Jagdale, Modified fracture properties of cement composites with nano/micro carbonized
bagasse fibers, Frat. Ed. Integrità Strutt. 9 (2015) 534–542, https://doi.org/10.3221/IGF-ESIS.34.59.
[41] K. Paine, Bacteria-based self-healing of concrete: effects of environment, exposure and crack size, Proc. RILEM Conf. Microorg. Mater. Interact. 1–15 (2016).
https://researchportal.bath.ac.uk/en/publications/bacteriabased-selfhealing-concrete-effects-of-environment-exposure-and-crack-size(dce79673-3376-4a8d-
a294-ab31a5dc0a6d).html.
[42] ASTM C31 ASTM C31, Standard practice for making and curing concrete test specimens in the laboratory, Annu. Book ASTM Stand. (2013) 1–8, https://doi.org/
10.1520/C0192.
[43] ASTM C1585-13, Standard test method for measurement of rate of absorption of water by hydraulic-cement concretes, ASTM Int 41 (2013) 1–6, https://doi.org/
10.1520/C1585-13.2.
[44] V. Shubina, L. Gaillet, T. Chaussadent, T. Meylheuc, J. Creus, Biomolecules as a sustainable protection against corrosion of reinforced carbon steel in concrete,
J. Clean. Prod. 112 (2016) 666–671, https://doi.org/10.1016/J.JCLEPRO.2015.07.124.
[45] G. Ramesh, E.D. Sotelino, W.F. Chen, Effect of transition zone on elastic moduli of concrete materials, Cement Concr. Res. 26 (1996) 611–622, https://doi.org/
10.1016/0008-8846(96)00016-6.
[46] S. Gupta, H.W. Kua, C.Y. Low, Use of biochar as carbon sequestering additive in cement mortar, Cem. Concr. Compos. 87 (2018) 110–129, https://doi.org/
10.1016/j.cemconcomp.2017.12.009.
[47] K. Vijay, M. Murmu, Self-repairing of concrete cracks by using bacteria and basalt fiber, 2019, SN Appl. Sci. 111 (1) (2019) 1–10, https://doi.org/10.1007/
S42452-019-1404-5.
[48] M. Luo, C. Qian, Influences of bacteria-based self-healing agents on cementitious materials hydration kinetics and compressive strength, Construct. Build. Mater.
121 (2016) 659–663, https://doi.org/10.1016/J.CONBUILDMAT.2016.06.075.
[49] R.A. Khushnood, S. Ahmad, P. Savi, J.M. Tulliani, M. Giorcelli, G.A. Ferro, Improvement in electromagnetic interference shielding effectiveness of cement
composites using carbonaceous nano/micro inerts, Construct. Build. Mater. 85 (2015) 208–216, https://doi.org/10.1016/J.CONBUILDMAT.2015.03.069.
[50] L. Restuccia, G.A. Ferro, Nanoparticles from food waste: a “green” future for traditional building materials, in: J. B, E.L.V. Saouma (Eds.), 9th Int. Conf. Fract.
Mech. Concr, . Concr. Struct., Berkeley, United States, 2016, pp. 1–6, https://doi.org/10.21012/FC9.276.
[51] R.A. Khushnood, S. ud din, N. Shaheen, S. Ahmad, F. Zarrar, Bio-inspired self-healing cementitious mortar using Bacillus subtilis immobilized on nano-/micro-
additives, J. Intell. Mater. Syst. Struct. 30 (2018) 3–15, https://doi.org/10.1177/1045389X18806401.
[52] N. Shaheen, R.A. Khushnood, W. Khaliq, H. Murtaza, R. Iqbal, M.H. Khan, Synthesis and characterization of bio-immobilized nano/micro inert and reactive
additives for feasibility investigation in self-healing concrete, Construct. Build. Mater. 226 (2019) 492–506, https://doi.org/10.1016/J.
CONBUILDMAT.2019.07.202.
[53] M. Zamani, S. Nikafshar, A. Mousa, A. Behnia, Bacteria encapsulation using synthesized polyurea for self-healing of cement paste, Construct. Build. Mater. 249
(2020), 118556, https://doi.org/10.1016/j.conbuildmat.2020.118556.
[54] R. Jakubovskis, A. Jankutė, J. Urbonavičius, V. Gribniak, Analysis of mechanical performance and durability of self-healing biological concrete, Construct.
Build. Mater. 260 (2020), https://doi.org/10.1016/j.conbuildmat.2020.119822.
[55] W.C. Choi, H. Do Yun, J.Y. Lee, Mechanical properties of mortar containing bio-char from pyrolysis, J. Korea Inst. Struct. Maint. Insp. 16 (2012) 67–74, https://
doi.org/10.11112/JKSMI.2012.16.3.067.
[56] A. Goldman, A. Bentur, The influence of microfillers on enhancement of concrete strength, Cement Concr. Res. 23 (1993) 962–972, https://doi.org/10.1016/
0008-8846(93)90050-J.
[57] M. Bouasker, P. Mounanga, P. Turcry, A. Loukili, A. Khelidj, Chemical shrinkage of cement pastes and mortars at very early age: effect of limestone filler and
granular inclusions, Cem. Concr. Compos. 30 (2008) 13–22, https://doi.org/10.1016/J.CEMCONCOMP.2007.06.004.
[58] A.M. Poppe, G. De Schutter, Cement hydration in the presence of high filler contents, Cement Concr. Res. 35 (2005) 2290–2299, https://doi.org/10.1016/J.
CEMCONRES.2005.03.008.
[59] A.R. Boğa, İ.B. Topçu, Influence of fly ash on corrosion resistance and chloride ion permeability of concrete, Construct. Build. Mater. 31 (2012) 258–264,
https://doi.org/10.1016/j.conbuildmat.2011.12.106.
[60] ASTM C597, Standard specification for pulse velocity through concrete, Annu. Book ASTM Stand. (2009) 6–9, https://doi.org/10.1520/C0597-09.2.
[61] K. Karhu, T. Mattila, I. Bergström, K. Regina, Biochar addition to agricultural soil increased CH4 uptake and water holding capacity - results from a short-term
pilot field study, Agric. Ecosyst. Environ. 140 (2011) 309–313, https://doi.org/10.1016/J.AGEE.2010.12.005.

18
M. Kanwal et al. Journal of Building Engineering 70 (2023) 105930

[62] W.A.W.A.K. Ghani, A. Mohd, G. da Silva, R.T. Bachmann, Y.H. Taufiq-Yap, U. Rashid, A.H. Al-Muhtaseb, Biochar production from waste rubber-wood-sawdust
and its potential use in C sequestration: chemical and physical characterization, Ind. Crop. Prod. 44 (2013) 18–24, https://doi.org/10.1016/J.
INDCROP.2012.10.017.
[63] S. Gupta, S.D. Pang, H.W. Kua, Autonomous healing in concrete by bio-based healing agents – a review, Construct. Build. Mater. 146 (2017) 419–428, https://
doi.org/10.1016/J.CONBUILDMAT.2017.04.111.
[64] B. Tasim, T. Masood, Z.A. Shah, M. Arif, A. Ullah, G. Miraj, uhammad Samiullah, Quality evaluation of biochar prepared from different agricultural residues,
Sarhad J. Agric. 35 (2019), https://doi.org/10.17582/journal.sja/2019/35.1.134.143.
[65] E. Ahmad, N. Jäger, A. Apfelbacher, R. Daschner, A. Hornung, K.K. Pant, Integrated thermo-catalytic reforming of residual sugarcane bagasse in a laboratory
scale reactor, Fuel Process. Technol. 171 (2018) 277–286, https://doi.org/10.1016/J.FUPROC.2017.11.020.
[66] L.C. Reimer, J. Sardà Carbasse, J. Koblitz, C. Ebeling, A. Podstawka, J. Overmann, BacDive in 2022: the knowledge base for standardized bacterial and archaeal
data, Nucleic Acids Res. 50 (2022) D741–D746, https://doi.org/10.1093/NAR/GKAB961.
[67] X. Hu, Y. Liu, D. Li, W. Feng, H. Ni, S. Cao, F. Lu, Y. Li, An innovative biotransformation to produce resveratrol by Bacillus safensis, RSC Adv. 9 (2019)
15448–15456, https://doi.org/10.1039/C9RA01338E.
[68] M. Sarkar, N. Alam, B. Chaudhuri, B. Chattopadhyay, S. Mandal, Development of an improved E. coli bacterial strain for green and sustainable concrete
technology, RSC Adv. 5 (2015) 32175–32182, https://doi.org/10.1039/C5RA02979A.
[69] W. De Muynck, N. De Belie, W. Verstraete, Microbial carbonate precipitation in construction materials: a review, Ecol. Eng. 36 (2010) 118–136, https://doi.org/
10.1016/J.ECOLENG.2009.02.006.
[70] G. Bin Cai, S.F. Chen, L. Liu, J. Jiang, H. Bin Yao, A.W. Xu, S.H. Yu, 1,3-Diamino-2-hydroxypropane-N,N,N′ ,N′ -tetraacetic acid stabilized amorphous calcium
carbonate: nucleation, transformation and crystal growth, CrystEngComm 12 (2009) 234–241, https://doi.org/10.1039/B911426M.
[71] S. Hajji, T. Turki, A. Boubakri, M. Ben Amor, N. Mzoughi, Study of Cadmium Adsorption onto Calcite Using Full Factorial Experiment Design, (n.d.). doi:
10.5004/dwt.2017.21079.
[72] F. Montes, S. Valavala, L.M. Haselbach, A new test method for porosity measurements of portland cement pervious concrete, J. ASTM Int. (JAI) 2 (2005),
https://doi.org/10.1520/JAI12931.
[73] S. Gupta, H.W. Kua, Effect of water entrainment by pre-soaked biochar particles on strength and permeability of cement mortar, Construct. Build. Mater. 159
(2018) 107–125, https://doi.org/10.1016/J.CONBUILDMAT.2017.10.095.
[74] Y. Yang, M.D. Lepech, E.-H. Yang, V.C. Li, Autogenous healing of engineered cementitious composites under wet–dry cycles, Cement Concr. Res. 39 (2009)
382–390, https://doi.org/10.1016/J.CEMCONRES.2009.01.013.
[75] T. Qureshi, A. Kanellopoulos, A. Al-Tabbaa, Autogenous self-healing of cement with expansive minerals-II: impact of age and the role of optimised expansive
minerals in healing performance, Construct. Build. Mater. 194 (2019) 266–275, https://doi.org/10.1016/j.conbuildmat.2018.11.027.
[76] X. Xian, C. Logan, Y. Shao, Dimensional stability of cement paste and concrete subject to early-age carbonation curing, Mater. Struct. Constr. 55 (2022) 1–19,
https://doi.org/10.1617/S11527-022-01926-8/FIGURES/9.
[77] W. Franus, R. Panek, M. Wdowin, Sem investigation of microstructures in hydration products of portland cement, Springer Proc. Phys. 164 (2015) 105–112,
https://doi.org/10.1007/978-3-319-16919-4_14/COVER.
[78] P. Ghods, O.B. Isgor, G.J.C. Carpenter, J. Li, G.A. McRae, G.P. Gu, Nano-scale study of passive films and chloride-induced depassivation of carbon steel rebar in
simulated concrete pore solutions using FIB/TEM, Cement Concr. Res. 47 (2013) 55–68, https://doi.org/10.1016/J.CEMCONRES.2013.01.009.
[79] Z. Wang, J. Yu, G. Li, M. Zhang, C.K.Y. Leung, Corrosion behavior of steel rebar embedded in hybrid CNTs-OH/polyvinyl alcohol modified concrete under
accelerated chloride attack, Cem. Concr. Compos. 100 (2019) 120–129, https://doi.org/10.1016/j.cemconcomp.2019.02.013.
[80] W. Feng, A. Tarakbay, S. Ali Memon, W. Tang, H. Cui, Methods of accelerating chloride-induced corrosion in steel-reinforced concrete: a comparative review,
Construct. Build. Mater. 289 (2021), 123165, https://doi.org/10.1016/j.conbuildmat.2021.123165.
[81] ASTM C 876-91, Standard test method for half-cell potentials of uncoated reinforcing steel in concrete, ASTM Int 91 (2015) 1–6, https://doi.org/10.1520/
C0876-09.2.
[82] L. Freire, M.A. Catarino, M.I. Godinho, M.J. Ferreira, M.G.S. Ferreira, A.M.P. Simões, M.F. Montemor, Electrochemical and analytical investigation of passive
films formed on stainless steels in alkaline media, Cem. Concr. Compos. 34 (2012) 1075–1081, https://doi.org/10.1016/J.CEMCONCOMP.2012.06.002.
[83] M. Cabrini, F. Fontana, S. Lorenzi, T. Pastore, S. Pellegrini, Effect of organic inhibitors on chloride corrosion of steel rebars in alkaline pore solution, 2015,
J. Chem. (2015), https://doi.org/10.1155/2015/521507.
[84] ATCC.org, Bacillus Safensis - BAA-1127, 2022. ATCC, https://www.atcc.org/products/baa-1127. (Accessed 9 December 2022). accessed.
[85] K.Y. Ann, H.S. Jung, H.S. Kim, S.S. Kim, H.Y. Moon, Effect of calcium nitrite-based corrosion inhibitor in preventing corrosion of embedded steel in concrete,
Cement Concr. Res. 36 (2006) 530–535, https://doi.org/10.1016/J.CEMCONRES.2005.09.003.
[86] Calcium nitrate = 99.0 13477-34-4, n.d. https://www.sigmaaldrich.com/PK/en/product/sigald/c1396. (Accessed 10 December 2022). accessed

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