Case Studies in Construction Materials 11 (2019) e00248

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Case Studies in Construction Materials

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Case study

Performance enhancement of brick aggregate concrete using

microbiologically induced calcite precipitation
Tanvir Manzura,*, Rafid Shams Huqb , Ikram Hasan Efazc , Sumaiya Afrozd,
Farzana Rahmane , Khandaker Hossainf
a
Ryerson University, Civil Engineering, Department of Civil Engineering, M5B 2K3, Toronto, ON, Canada
b
Stamford University Bangladesh
c
University of Texas at Arlington, Arlington
d
Bangladesh University of Engineering & Technology
e
University of Texas at Austin
f
Ryerson University

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

Article history: The use of brick aggregates is very common in Bangladesh and nearby regions as artificial
Received 1 January 2019 aggregate replacing natural stone. To ensure sustainable construction, finding a way to
Received in revised form 9 May 2019 enhance its durability is extremely necessary. Microbiologically Induced Calcite
Accepted 9 May 2019
Precipitation (MICP) using indigenous urease positive bacteria collected from soil showed
promising results in initial research, hence further investigation has been carried out for
Keywords: comprehensive understanding on the effectiveness of this method. Two incubation periods
Brick aggregates
of 24 hours and 48 hours were considered for bacterial culture. Absorptiometric method
Durability
Incubation
was used to examine population growth of bacteria due to varying incubation period. Both
Bacteria Scanning Electron Microscopy (SEM) and Energy Dispersive Spectroscopy (EDS) were used
MICP to identify the presence of CaCO3 due to bio-treatment. In addition to Rapid Migration Test
(RMT), Rapid Chloride Penetration Test (RCPT) was conducted to evaluate reduction of
concrete permeability due to application of treated brick aggregate. It has been found that
48 hours incubation was more effective than 24 hours incubation. The longer incubation
resulted in almost twofold reduction in aggregate absorption test. Compressive Strength,
RCPT and RMT of brick aggregate concrete resulting from 48 hours incubation showed 14%,
27% and 43% improvement over untreated brick concrete, respectively.
© 2019 The Authors. Published by Elsevier Ltd. This is an open access article under the CC
BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

1. Introduction

Natural resources are running out at a rate much higher than ever before. High growth rates of global economy spurred by
rapid industrialization have escalated the process. The growth of the construction sector is an inevitable component of
industrial boom and a source of depletion of natural resources. The construction sector has a significant impact on the
environment [1]. Annually, building construction in the world consumes: 25% of the global wood harvest; 40% of stone, sand
and gravel; and 16% of water [2]. An annual demand of 53.2 billion metric tonnes of stone is expected to be required through
2017 [3]. However, due to unavailability and high price, stones are replaced by inexpensive and more widely available
materials, such as brick aggregates, in some developing countries for certain types of construction.

* Corresponding author.
E-mail address: tmanzur@ryerson.ca (T. Manzur).

https://doi.org/10.1016/j.cscm.2019.e00248
2214-5095/© 2019 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/
licenses/by-nc-nd/4.0/).
2 T. Manzur et al. / Case Studies in Construction Materials 11 (2019) e00248

Aggregates obtained from crushing of burnt clay bricks are particularly popular in number of countries in the world like
Bangladesh where natural source of stone aggregates is scarce. This is because it is relatively inexpensive and widely available.
Although widely used, concrete made using brick aggregates are weaker and proven to have lower durability than those made
using stone chips [4–12]. This shortcoming of brick aggregates arises from their inherent porous structures which make
moisture penetration and micro-crack propagation much easier. Nevertheless, in the absence of a practical substitute for brick
aggregates in countries with limited access to natural stone aggregates, it is important to improve the durability of concretes
made with brick aggregates. Microbially Induced Calcite Precipitation (MICP) is a process which has been successfully employed
by various researchers to make improvements at materials scale. The idea of using microbes to be applied to bio-geotechnical
engineering was first introduced in 1992 [13]. Later, further research was carried out to extend the possibility of employing
microbes in the geotechnical field as one of eco-friendly technologies [14–18]. The innovative technique of remediating cracks in
concrete using calcite precipitation by MICP was proposed by Ramakrishnan et al. [19]. The process is considered as a sustainable
and environmentally-friendly method for crack remediation in concrete since it does not deplete natural resources [20], it is
pollution free [21] and does not emit any greenhouse gases [22]. It has been successfully employed in cementing cracks in granite
[23], stone conservation [24], concrete [25], masonry brick, building structures, pavements, and restoration of historic
monuments [26]. It has also been used for enhancement of construction materials like sand [27–30], improving the water
absorbing capacity of whole bricks [31], improving the durability of cohesionless soils [32] and also improved the performance
of concrete by increasing strength and durability [33]. Inspired by the success of previous researches, a research initiative has
been undertaken at Bangladesh University of Engineering and Technology (BUET) to apply MICP technique for filling the pores of
brick aggregates and hence, improving the durability of concrete incorporated with them as a case study. This article presents the
outcome of the 2nd phase of the study conducted. The initial findings of the study are available in the article by Manzur et al. [34].
The initial phase of research was based on 24 h incubation period for bacterial growth. Various tests like optical microscopy,
scanning electron microscopy (SEM), X-ray powder diffraction (XRD), and energy-dispersive spectroscopy (EDS) conducted on
brick aggregates showed biodepostion of CaCO3 on brick surfaces. Decreased absorption of treated brick aggregate and
enhanced durability performance of resultant concrete were also observed. However, since the application of MICP for
enhancement of brick aggregate and brick aggregate concrete is currently at inception stage, further research results obtained
through different testing procedure is required to consolidate the findings of the initial phase of research. Hence, in the second
phase, two incubation periods of 24 h and 48 h were used to investigate the effect of incubation on effectiveness of MICP in
enhancing brick aggregate properties. In addition to Scanning Electron Microscope (SEM) and Energy Dispersive Spectroscopy
(EDS), absorptiometric method using a HACH Spectrophotometer was used in this phase to examine and compare the
population growth of bacteria for different incubation periods. After confirmation of calcite precipitation in each specimen,
absorption tests were conducted. The durability of concrete, made with treated brick aggregates, was evaluated through Rapid
Chloride Penetration Test (RCPT) and Rapid Migration Test (RMT). The previous study [34] was based on RMT only. All these test
results are vital to encourage further research in this field.

2. Materials used and methodology

2.1. Isolation of urease positive Bacteria and microbial culture

Highly urease positive bacteria Sporosarcinapasteurii (formerly known as Bacillus pasteurii) areusually used in the study of
application of MICP technique in soil and cementitious composites [33,35–38]. Sporosarcinapasteurii has the ability to
precipitate calcite efficiently in the presence of a calcium source and urea. However, in Bangladesh, Sporosarcinapasteurii is
not readily available. Thus, it was decided to isolate urease positive bacteria from the surrounding environment following the
procedures described in the previous studies [28,34]. However, only the most rapid urease positive strains were used and
they were identified by Motility Indole Urease (MIU). Bacteria concentration is usually higher at the top surface of soil.
Therefore, bacteria were isolated from the top surface of the garden soil where urea fertilizer has been used frequently. In the
first cultivation, soil was collected before monsoon. A 225 ml autoclaved (121  C and 15 psi for 15–20 min) conical flask was
filled with 50 ml of autoclaved (121  C and 15 psi for 15–20 min) ammonium sulphate–yeast extract solution (NH4-YE) broth
medium. The medium was inoculated with 5gm of soil. After 2 days of incubation at 30  C, one loopful was taken from the
broth and streaked into a NH4-YE agar plate medium. This plate was incubated for one day at 30  C. Mixed culture of bacteria
was grown in the plate. After that, several trial bacteria colonies were streaked into several plate media and each plate was
incubated for one day at 30  C. This process was done to get the purest strain of bacterium. Heavy inoculant from each plate
medium was inoculated in motility indole urease (MIU) medium in the test tube for urease positive test. The presence of
urease enzyme in bacteria is indicated by change of color of the MIU medium from yellow to pink. The most rapid urease
positive bacteria were identified from the lowest possible time required to change the mentioned color. Finally, the identified
bacteria strain was cultured in conical flask containing 150 ml of culture medium for bio-treatment solution. The entire
process is presented through a flowchart in Fig. 1.

2.2. Material used

Ordinary Portland cement (OPC) with unit weight of 3150 kg/m3 as the main binder and crushed clay bricks as coarse
aggregates were used in this study. The main constituents of cement, obtained throughX-ray Fluorescence (XRF) analysis, are
 T. Manzur et al. / Case Studies in Construction Materials 11 (2019) e00248 3

Fig. 1. Flow chart describing the isolation of Urease Positive Bacteria from soil.

provided in Table 1. The bricks used were well shaped, reddish in color and had unit weight and strength of 1050 kg/m3and
21.5 MPa, respectively. The gradation of brick aggregate was measured following ASTM C136 [39] standard and is shown in
Fig. 2. Locally available river sand known as “Sylhet sand” was used as fine aggregates. Fine aggregates were used with a
fineness modulus (FM) of 2.5 and unit weight of 1590 kg/m3.

2.3. Mix design of concrete

The concrete mix was prepared following the volumetric mix ratio of 1:1.5:3 (corresponding to cement: fine aggregates:
coarse aggregates, respectively) with water to cement ratio of 0.45. This mix ratio is quite common in local construction sites
as found from previous study [11] and was followed to imitate the local practice. Details of mix design are provided in Table 2.
Water sprinkling was done on all aggregates (treated and untreated) prior to mixing 24 h ahead to simulate the field practice
4 T. Manzur et al. / Case Studies in Construction Materials 11 (2019) e00248

Table 1
Mass percentage of main constituents of OPC.

Oxide Amount in percent

CaO 66
SiO2 18
Al2O3 3.0
Fe2O3 3.9
MgO 2.2
SO3 4.5
Na2O 0.20
K2O 1.2

Fig. 2. Gradation curve of brick aggregates.

carried out in the local construction sites. This would place the aggregates in a surface dry condition. In the general
construction fields where brick is used as coarse aggregate, no water correction is usually made after sprinkling of water. As a
result, brick aggregates remain in surface dry condition and their absorption capacity is not fully exhausted. Consequently,
resulting brick aggregate concrete suffers from high absorption of aggregate and show less strength and significantly high
permeability. Similar condition was simulated in this study. However, fixed amount of water was sprinkled for both treated
and untreated aggregates to maintain similar condition for comparison. The test specimens were cast in steel molds and
compacted with a vibrator nozzle. Specimens were casted and left for 24 h at atmospheric conditions (temperature of 23  C)
before demolding. After demolding, the specimens were completely submerged under lime water at temperature of 23  C.

2.4. Incorporation of Bacteria in brick aggregates

Bacteria incorporation within aggregate was done following the cyclic method suggested by Manzur et al. [34]. The entire
process is depicted through a flow chart as shown in Fig. 3. In total, 3 cycles of cementitious solutions were applied. The
cementitious solutions were prepared by mixing 140 ml of CaCl2.2H2O stock solution with 400 ml of urea nutrient medium
[28,34]. After each cycle, the solution was drained and new solution was poured. Eventually, bacteria settled on the brick
chips with time. Overall, the total duration of microbial treatment was 21 days excluding initial 3 days of microbial
attachment. The specimens were fully submerged under solution for both 24 h and 48 h incubation variations.

2.5. Bio-Treatment Method (incubated for 24 h and 48 h)

The bio-treatment procedures followed in this study were proposed by Manzur et al. [34]. NH4-YE broth medium was
taken in to Duran bottles inside the hood. The initial portion of the opening of the bottles was burnt for 2-3 seconds and it
was closed with the cotton plug. One loopful of bacteria was taken from the culture plate and inoculated into the medium
inside the hood at 30  C for two incubation periods. Then urea nutrient medium was taken into container. CaCl2.2H2O stock
solution and all the NH4-YE broth medium with bacteria (which was incubated for 24 and 48 h at 30  C) were added into urea

Table 2
Mix design details.

Water Cement Ratio Cement (Kg/m3) Water (Kg/m3) FA (Kg/m3) CA (Kg/m3)

0.45 419 188.6 685 977
 T. Manzur et al. / Case Studies in Construction Materials 11 (2019) e00248 5

Fig. 3. Flow chart describing the process of bacteria incorporation.

nutrient solution. These solutions were then poured on the aggregates. This initial biological treatment was allowed to set
three days to allow the microbes to attach to the sample. After the initial biological treatment of three days, the solutions
with suspended bacteria were drained and three cycles of cementation treatment solution were applied as discussed in the
previous section. The bio-treatment method is described through flowchart in Fig. 4. The visual change to whitish
appearance of the treated aggregates as compared to the stark red color of the control is shown in Fig. 5.

2.6. Tests conducted on bio-modified brick aggregate

A number of tests were performed in order to validate calcium carbonate precipitation and performance improvements.
Scanning electron microscopy (SEM) and EDS analysis of brick chips were conducted to ensure calcium carbonate
precipitation. In addition, absorption test, Rapid Migration Test (RMT) and Rapid Chloride Permeability Test (RCPT) were also
done as durability test of concrete. Compressive strength of treated brick aggregate concrete and control concrete were also
measured and compared.

2.7. Measurement of population of bacteria in solutions by Absorptiometric Method

After NH4-YE broth medium was taken in to Duran bottle, its absorbance was measured by Absorptiometric Method using
a HACH Spectrophotometer (Fig. 6) with a wavelength of 590 nm [40]. First, the sample cell was filled with 10 ml of deionized
water which is called blank. The blank was inserted into the cell holder to calibrate the spectrophotometer. After calibration,
10 ml of NH4-YE broth medium was poured in the sample cell and the sample cell was inserted into the cell holder. The result
shown was the absorbance of the NH4-YE broth medium without bacteria. Two sets of NH4-YE broth medium were prepared
6 T. Manzur et al. / Case Studies in Construction Materials 11 (2019) e00248

Fig. 4. Flow chart describing the process of biotreatment.

Fig. 5. (a) Control aggregate; (b) treated aggregate (24 h incubation); (c) treated aggregate (48 h incubation).

and one loopful of bacteria was inoculated in each set. Then the mediums were incubated at 30  C for 24 h and 48 h,
respectively. After incubation with bacteria, absorbance of both solutions was measured again.

2.8. Compressive strength test of concrete specimens

The compressive strength test was performed according to ASTM C39 [41]. Concrete cylinders with diameter of 100 mm
and height of 200 mm were casted and then placed in the curing pond. In the curing pond, the specimens were completely
submerged under lime water. The temperature of lime water was 23  C in the pond. After proper curing, compressive
strength of each variation was measured by crushing the cylinders. An axial load with a rate of 440 lb/sec was applied until
failure. Three specimens were used for each variation.
 T. Manzur et al. / Case Studies in Construction Materials 11 (2019) e00248 7

Fig. 6. HACH Spectrophotometer.

2.9. Rapid Migration Test (RMT)

Rapid Migration Test was conducted in accordance with AASHTO TP 64-03 [42] on specimens cured for 60 days. RMT
involves subjecting 100 mm diameter and 50 mm thick cylindrical specimen to an external electric potential to force
chloride ions to migrate into the specimen. To account for varying concrete resistance, the initial current flow through the
specimen was measured and applied voltage was adjusted accordingly. The samples were fit into silicon rubber sleeves
where one side of the specimen was immersed in a 1.2% NaOH solution and the other side to a 10% NaCl solution. After 18 h,
the samples were removed and axially split into two pieces. The depth of chloride penetration was determined in one half
of the specimen using a colorimetric technique, spraying silver nitrate solution on the freshly cut surface. The penetration
depths were measured discarding the highest and lowest values and taking the average of 5 penetration depths. The rate of
penetration was calculated by dividing the depth of penetration (mm) by the product of the applied voltage (V) and the
test duration (hr).

2.10. Rapid Chloride Permeability Test (RCPT)

Standardized testing procedures of RCPT are covered in AASHTO T 277 [43] or ASTM C 1202 [44] and have been of interest
to numerous papers in the open literature [45–49]. Samples, cured for 60 days, were tested for chloride permeability
following the above mentioned codes and using the setup developed by Iffat et al. [50]. Similar to RMT, cylindrical specimens
with diameter 100 mm and a thickness of 50 mm were sliced from cast cylinder, after being removed from curing ponds. The
excess water was wiped from the specimen surfaces and after that side surfaces were painted to make them water tight.
Once the paint dried, the surface-dried specimens were placed in the vacuum desiccator, for preconditioning, with both end
surfaces exposed. The vacuum pump was turned on and the absolute pressure in the desiccator was decreased to around
1 mm Hg and this vacuum was maintained for three hours. Without turning off the pump, the water stopcock was opened
and the desiccator was filled with de-aired tap water to submerge the concrete specimens. Closing the water stopcock, the
vacuum was maintained for additional one hour. After the additional one hour, the pump was switched off and pressure was
released to allow air to re-enter the desiccator. The submerged specimens were allowed to remain as they were inside the
desiccator for 18 h. The RCPT was performed by monitoring the amount of electrical current that passed through a sample
50 mm thick by 100 mm in diameter in 6 h. One end was immersed in a 3.0% salt (NaCl) solution and the other in a 0.3 M
sodium hydroxide (NaOH) solution.

3. Results and discussions

3.1. Confirming presence of bacterial growth

Concentration of bacteria was measured using HACH Spectrophotometer with a wavelength of 590 nm. The wave length
of 590 nm was used since many bacterial growth media contain yellowish-brown compounds that absorb at the shorter
wavelengths. It is common method for estimating the concentration of bacterial or other cells in a liquid. It is also called
OD600 which is basically the absorbance or optical density of a sample measured at a wavelength of 600 nm (in this case it
was 590 nm). The increase of absorbance after incubating the solution indicated the growth of bacteria in the solution. The
initial absorbance before bacterial incubation was 0.05 ppb (parts per billion) which increased to 1.03 ppb and 1.64 ppb for
24 h and 48 h specimen, respectively. This indicates that the bacterial concentration increased for both specimens, although
the increase for 48 h sample was much higher representing the greater increase of bacterial concentration. The results are
consistent with other research works on increase in concentration of bacteria due to increased incubation period [51].
8 T. Manzur et al. / Case Studies in Construction Materials 11 (2019) e00248

3.2. Confirmation of calcium carbonate precipitation

In order to analyze the effectiveness of MICP and effect of varying the bacterial incubation periods, it was necessary to
confirm that calcium carbonate precipitation occurred as intended. Analysis of the results from SEM, optical Spectroscopy
and energy dispersive spectroscopy confirmed that calcium carbonate precipitation was indeed successful.

3.2.1. Scanning Electron microscopy (SEM) and optical spectroscopy

It is distinctly observable that the two photographs in Fig. 7, although of the same magnification, are different from each
other. In the SEM image of the control sample i.e. (a) of Fig. 7, the topography is very much smooth, but in the SEM image of
the treated sample i.e. (b) of Fig. 7, spherical nodules can be observed. Images of higher magnification in Fig. 8 also show that
spherical nodules are present on both 24 -h and 48 -h-incubated samples. Urease-positive bacteria perform urea hydrolysis
to form ammonia and carbon dioxide, which in turn aid to create alkaline conditions. With increase in alkalinity, calcite
precipitation is facilitated with the availability of calcium cations in the near surrounding. As a result, such spherical nodules
were observed. The chemical reaction leading to formation of calcium carbonate is as follows:
Ca2+ + HCO3 + OH → CaCO3(s) + H2O
Also, optical microscopy images as shown in Fig. 9 validate the presence of white precipitated object only on the treated
specimens. Similar observations were also made in the initial phase of research and powder x-ray diffractometry of treated
brick aggregate identified the crystalline phase of precipitated objects as vaterite [34]. No vaterite peak was found in
untreated brick aggregates. Two conclusions can be made from the images. Firstly, MICP technique using locally available
urease positive bacteria does precipitate vaterite (CaCO3) on the brick aggregates and secondly, the deposition of mineral is
not uniform as the process is part of bacterial metabolic activity which is random in nature.

3.2.2. Energy dispersive spectroscopy

The EDS results of control and 24 -h incubated specimen are shown in Fig. 10. The mass percentages of various pertinent
elements found from EDS analysis of control, 24 h and 48 h specimens are provided in Table 3. In the 24 -h specimen, the
percent mass of calcium and carbon was found as 17.81% and 22.59%, a significant increase from 0.53% and 2.75%,

Fig. 7. SEM micrographs of (a) control aggregates; (b) 24 h treated aggregates.

Fig. 8. SEM micrographs of (a) 24 h and (b) 48 h samples.

 T. Manzur et al. / Case Studies in Construction Materials 11 (2019) e00248 9

Fig. 9. Optical microscopy images (a) Control aggregates (b) 24 h treated aggregates.

Fig. 10. EDS analysis results of specimen: (a) control; (b) 24 h specimen.

respectively from control sample. This increase was due to precipitation of calcium carbonate on the brick aggregates by
MICP. For 48 -h specimen, the respective percent masses of calcium and carbon were 38.07% and 15.64%, respectively. The
percent mass of calcium increased further since the precipitation in 48 h sample was higher than 24 h sample.

3.3. Effect of MICP on brick aggregates and concrete

3.3.1. Absorption test

The bar charts in Fig.11 represent the amount of water absorbed by the three specimens. It is quite evident that the amount of
water absorbed decreased due to MICP treatment. Around 6% and 18% reduction in water absorption by the treated brick
aggregates in the 24 h specimen and the 48 h specimen, respectively were obtained. This impressive reduction in water
absorptionwas due to the precipitated calcium carbonates filling up the pores of the brick aggregates. Such pore sealing hindered
the penetration of water into the aggregates and consequently, enhanced the water resistivity. The EDS analysis showed that 48 h
sample had significant higher percentage of calcium. The higher incubation period facilitated higher precipitation and hence,
more effectively sealed the pores of the brick aggregates, leading to higher reduction in water absorption. It is to be noted that
48 h sample showed 14% absorption which is still quite high. Hence, higher incubation or using different species of bacteria may
reduce the absorption of brick aggregate and needs to be investigated in future studies.

3.3.2. Compressive strength

The main objective of the research was to improve concrete durability through reducing permeability of brick aggregate.
However, reduction in absorption of brick aggregate was also expected to positively impact the compressive strength of
10 T. Manzur et al. / Case Studies in Construction Materials 11 (2019) e00248

Table 3
Mass percentage of different elements obtained from EDS analysis.

Element Mass % of control specimen Mass % of 24 h specimen Mass % of 48 h specimen

C 2.75 22.59 15.64
O 61.72 57.90 28.77
Al 1.94 0.05 1.69
Si 31.61 0.25 3.30
Cl 0.10 1.40 12.54
Ca 0.53 17.81 38.07

Fig. 11. Absorption by control and treated brick aggregates.

concrete. Fig. 12 shows the mean compressive strengths of 3 cylindrical samples from each variation. The error bars in Fig. 12
represents the standard deviation of strength test results of each sample variation. As observed in Fig. 12, the compressive
strength increased in both 24 -h specimen and 48 h specimen when compared with the control. The percent increases were
respectively, around 12% and 14%. The sealing of pores by precipitated calcium carbonate reduced water absorption by the
brick aggregates and thereby, ensured availability of required amount of water for hydration of cement. Brick aggregates
have high water absorption than natural stone aggregates. No moisture correction was done for the aggregates used. Since
the aggregates were at surface dried condition, the moisture content of the aggregates was lower than the absorption
capacity. Hence, the brick aggregates would pull away water from the batch mixture. The sealing off of the pores decreased
the water absorption capacity of the treated brick aggregates. Thus less of the batch water would be pulled away by the
treated aggregates and more would be available for cement hydration.

3.3.3. Durability test results of concrete

3.3.3.1. Rapid Migration Test (RMT). As observed in Fig. 13, rate of penetration decreased in both 24 h and 48 h specimens in
comparison to the control. The respective percentages reductions in rate of penetration were 14.3% and 42.9%. It is conclusive
from the results that the pores of brick aggregates were sealed due to MICP. Such sealing resulted in better hydration and as
well as hindered penetration of water through brick aggregate. As a result, reduction in the rate of penetration by chloride
ions was obtained. The results of RMT can be visually observed from the surfaces of the split samples of the concrete
cylinders sprayed with silver nitrate solution. This spraying led to the formation of white colored silver chloride representing
zones of successful chloride penetration and brown silver hydroxide where penetration was not successful. It is observable in
Fig. 14 (a) that the depth of penetration is very deep. The highest depth of penetration in control samples was up to 50 mm
and average penetration was also close to 50 mm. Most of the area is white in color, indicating successful penetration within
the control specimen. In case of 24 h specimen, depth of penetration was at no place more than 40 mm and the average
penetration was much lower (as seen in Fig. 14 (b)), indicating the reduced penetration through the 24 h specimen. Hence,
porosity of concrete has been successfully reduced.

3.3.3.2. Rapid Chloride Penetration Test (RCPT). It is quite evident from Fig. 15 that the amount of charged passed is reduced in
both 24 h and 48 h treated specimens in comparison with the control specimen. In comparison to the control, a percentage
reduction of 7.0% and 27.3% was observed, respectively. In the current vs time graph shown in Fig. 16, it is clear that there was
a steady increase of current before levelling off after a certain point of time. The increase of current was highest for the
control and the least for 48 -h specimen. The 24 h specimen seems to be in between the two extremes throughout most of the
experiment periods. Hence, it is obvious that MICP treatment of brick aggregate resulted in robust concrete due to enhanced
hydration and penetration resistance of brick chips resulting from precipitation of calcium carbonates.
 T. Manzur et al. / Case Studies in Construction Materials 11 (2019) e00248 11

Fig. 12. Results of compressive strength tests.

Fig. 13. RMT test results.

Fig. 14. Measurement of chloride penetration depth on split face of (a) untreated brick aggregate. concrete (Control); (b) 24 h treated brick aggregate
concrete.

It is apparent from strength, RMT and RCPT tests that greater improvement in permeability was observed as compared to
the compressive strength test. This is due to the fact that MICP technique improves the microstructure of concrete by sealing
the pores in the aggregate which imparts in the increased durability. The water can penetrate through the brick aggregate
due to its high porosity and therefore, a continuous path may become available through cement paste and brick aggregate.
However, treated aggregate showed significant improvement in permeability as observed from absorption test. Moreover,
the previous research by Manzur et al. [34] ensured the significant reduction in permeability of the aggregate by conducting
the mercury intrusion porosimetry (MIP) test on MICP treated sample. But the phase of C-S-H (calcium silicate hydrate)
probably did not change due to reduction in permeability of aggregate which is the main strength giving component in a set
of cement paste. That is a probable reason of achieving higher durability but lower enhancement of mechanical strength for
MICP treated aggregate concrete. The lower increase of mechanical strength is probably due to the increased density of
microstructure because of CaCO3 deposition.
12 T. Manzur et al. / Case Studies in Construction Materials 11 (2019) e00248

Fig. 15. RCPT Results (Charged Passed).

Fig. 16. Current Vs Time Graph from RCPT.

4. Conclusions

It can be concluded that bio-treatment of brick aggregate through MICP is an effective method in enhancing durability
properties of resultant concrete and that increasing incubation period from 24 h to 48 h has a significant improvement in
performance. Results were collected for control, 24 h and 48 h samples and compared to each other to make the following
conclusions:

 The incubation period has effect on the population of bacteria. The absorbance of the broth before and after inoculation of
bacteria confirmed that the population of bacteria increased due to increased incubation period from 24 h to 48 h. The
increased number of bacteria improved the efficiency of the technique as observed from other test results.
 On visual inspection, a white deposition was observed on the MICP treated brick aggregates (24 h and 48 h specimen). The
deposition was further confirmed by SEM imaging.
 From EDS, it was observed that the percentage of both Calcium and Carbon increased for MICP treated specimen than that
of control ones. Hence, strongly indicating the deposition is that of CaCO3.
 The percent reduction in water absorption of brick aggregate was about three times more effective with 48 h incubation
method than that of 24 h incubation. Undoubtedly, increase in incubation period increased the efficiency of MICP
treatment and facilitated greater reduction in water absorption.
 MICP treated brick aggregate resulted in moderate improvement of concrete strength as expected.
 Enhanced durability performance of brick aggregate concrete can be achieved through MICP. The RMT results showed that
the rate of penetration decreased by 14% in 24 -h specimen and this improved to 43% reduction in the 48 -h specimen.
 The RCPT results further consolidated the effectiveness of MICP as a mean of improving durability and confirmed that
increasing incubation period improves the durability enhancement even more. The amount of current passed was reduced
by 7% and 27% in 24 h specimen and 48 h specimen, respectively compared to control specimens.
 T. Manzur et al. / Case Studies in Construction Materials 11 (2019) e00248 13

Although, the research is in its early stage, the results are certainly encouraging. MICP can be the solution that is required
by the local construction industry to mitigate the drawbacks of using brick aggregate for concrete. Also, this might encourage
other countries to adopt this solution as a means of reducing the depletion of natural resources i.e. stones. However, much
work is yet to be done in order to establish MICP as commercially viable and reliable means of improving durability of brick
aggregate concrete.

Conflict of interest statement

The authors of the manuscript titled “Performance Enhancement of Brick Aggregate Concrete using Microbiologically
Induced Calcite Precipitation” declare no conflict of interest of any of the authors with any organization in the subject matter
discussed in the manuscript.

Acknowledgments

The authors would like to acknowledge the officials and staffs of the Concrete and the Environment Laboratory of the Civil
Engineering Department of Bangladesh University of Engineering and Technology (BUET) for successful completion of this
study. The authors are also thankful to the Department of Materials and Metallurgical Engineering and the Department of
Glass and Ceramic Engineering of BUET for optical microscopy and scanning electron microscopy (SEM) images, respectively.

References

[1] L.F. Cabeza, L. Rincón, V.G. Vilariño, G. Pérez, A. Castell, Life cycle assessment (LCA) and life cycle energy analysis (LCEA) of buildings and the building
sector: a review, Renewable Sustainable Energy Rev. 29 (2014) 394–416.
[2] P. Joseph, S. Tretsiakova-McNally, Sustainable non-metallic building materials, Sustainability 2 (2010) 400–427.
[3] M.S. Kuhar, World Aggregates Market April, (2017) . http://www.rockproducts.com/features/13045.
[4] T. Manzur, B. Baten, M.J. Hasan, H. Akter, A. Tahsin, K.M.A. Hossain, Corrosion behavior of concrete mixes with masonry chips as coarse aggregate,
Constr. Build. Mater. 185 (2018) 20–29.
[5] B. Baten, M.J. Hasan, B. Bose, T. Manzur, Corrosion potential of Stone and brick aggregate concrete in the context of Bangladesh, Proceedings of 1st
Bangladesh Civil Engineering Summit, BUET, Dhaka, Bangladesh, (2016) .
[6] H. Akter, A. Tahsin, B. Bose, T. Manzur, Performance of brick aggregate concrete: Bangladesh context, Proceedings of the 1st Bangladesh Civil
Engineering Summit, BUET, Dhaka, Bangladesh, (2016) .
[7] A.R. Khaloo, Properties of concrete using crushed clinker brick as coarse aggregate, Aci Mater. J. 91 (4) (1994) 401–407.
[8] S. Afroz, F. Rahman, Influence of Bio-treated Bricks on Concrete Durability. Thesis (BSc), Bangladesh University of Engineering and Technology, 2015.
[9] S.Z. Bosunia, J.R. Chowdhury, Durability of concrete in coastal areas of Bangladesh, J. Civil Eng. IEB 29 (1) (2001) 41–53.
[10] I.H. Efaz, R.S. Huq, Effect of Incubation Period on Bio-treated Brick Concrete. Thesis (BSc), Bangladesh University of Engineering and Technology, 2016.
[11] S. Afroz, F. Rahman, S. Iffat, T. Manzur, Sorptivity and strength characteristics of commonly used concrete mixes of Bangladesh, Proceedings of the
Recent Innovation in Civil Engineering for Sustainable Development (IICSD), DUET, Gazipur, Bangladesh, (2015) .
[12] S. Afroz, F. Rahman, T. Manzur, A simple laboratory process to culture indigenous urease positive bacteria for MICP, Proceedings of the 1st Bangladesh
Civil Engineering Summit, BUET, Dhaka, Bangladesh, (2016) .
[13] F.G. Ferris, L.G. Stehmeier, Bacteriogenic mineral plugging, J. Can. Pet. Technol. (1992) 36.
[14] J.K. Mitchell, J.C. Santamarina, Biological considerations in geotechnical engineering, J. Geotech. Geoenvironmental Eng. 131 (2005) 1222–1233.
[15] J.T. DeJong, M.B. Fritzges, K. Nüsslein, Microbially induced cementation to control sand response to undrained shear, J. Geotech. Geoenvironmental Eng.
132 (2006) 1381–1392.
[16] V.S. Whiffin, L.A.V. Paassen, M.P. Harkes, Microbial carbonate precipitation as a soil improvement technique, Geomicrobiol. J. 24 (2007) 417–423.
[17] J. Chu, V. Ivanov, M. Naeimi, V. Stabnikov, H.L. Liu, Optimization of calcium-based bioclogging and biocementation of sand, ActaGeotechnica. 9 (2014) 277–285.
[18] J. Chu, V. Stabnikov, V. Ivanov, Microbially induced calcium carbonate precipitation on surface or in the bulk of soil, Geomicrobiol. J. 29 (2012) 544–549.
[19] V. Ramakrishnan, R. Panchalan, S.S. Bang, Bacterial concrete: a self remediating biomaterial, Proceedings of the 10th Int. Congress on the Polymers in
Concrete, ICPIC, Hawaii, (2001) .
[20] H.S. Patil, D.B. Raijiwala, H. Prashant, B. Vijay, Bacterial concrete: a self healing concrete, Int. J. Appl. Eng. Res. 3 (12) (2008) 1719–1725.
[21] S.K. Ramakrishnan, R. Panchalan, S.S. Bang, R. City, Improvement of concrete durability by bacterial mineral precipitation, Proceedings of the 11th Int.
Conf. on Fracture, Curran Associates, New York, (2005) , pp. 20–25.
[22] I.M. Isha, H. Afifudin, H. MohdSaman, Bacillus subtilisandthermusthermophilus: derived bioconcrete in enhancing concrete compressive strength, Int.
Sustainability Civ. Eng. J. 1 (1) (2012) 48–56.
[23] U.K. Gollapudi, C.L. Knutson, S.S. Bang, M.R. Islam, A new method for controlling leaching through permeable channels, Chemosphere 30 (4) (1995)
695–705.
[24] C. Rodriguez-Navarro, F. Jroundi, M. Schiro, E. Ruiz-Agudo, M.T. González-Muñoz, Influence of substrate mineralogy on bacterial mineralization of
calcium carbonate: implications for stone conservation, Appl. Environ. Microbiol. 78 (11) (2012) 4017–4029.
[25] C. Qian, R. Wang, L. Cheng, J. Wang, Theory of microbial carbonate precipitation and its application in restoration of cement-based materials defects,
Chin. J. Chem. 28 (5) (2010) 847–857.
[26] F. Cappitelli, L. Toniolo, A. Sansonetti, D. Gulotta, G. Ranalli, E. Zanardini, C. Sorlini, Advantages of using microbial technology over traditional chemical
technology in removal of black crusts from stone surfaces of historical monuments, Appl. Environ. Microbiol. 73 (17) (2007) 5671–5675.
[27] G.G.N.N. Amarakoon, S. Kawasaki, Factors affecting sand solidification using MICP with pararhodobacter sp, Mater. Trans. 59 (1) (2018) 72–81.
[28] F. Alam, Microorganism in Mitigating Seismic Liquefaction. Thesis (MSc), Bangladesh University of Engineering and Technology, 2014.
[29] A. Mukherjee, N.K. Dhami, B.V.V. Reddy, M.S. Reddy, Bacterial calcification for enhancing performance of low embodied energy soil-cement bricks,
Proceedings of the 3rd International Conference on Sustainable Construction Materials and Technologies, Kyoto, Japan, (2013) .
[30] A. Kantzas, L. Stehmeier, D.F. Marentette, F.G. Ferris, K.N. Jha, F.M. Maurits, A novel method of sand consolidation through bacteriogenic mineral
plugging, Annual Technical Meeting, Petroleum Society of Canada, (1992) .
[31] D. Sarda, H.S. Choonia, D.D. Sarode, S.S. Lele, Biocalcification by Bacillus pasteurii urease: a novel application, J. Ind. Microbiol. Biotechnol. 36 (8) (2009)
1111–1115.
[32] M. Kasinathan, S.S. Bettadapura, Durability of Microbially Induced Calcite Precipitation (Micp) Treated Cohesionless Soils, vol. 2, Japanese Geotechnical
Society Special Publication, 2016, pp. 1946–1949.
[33] P. Ghosh, S. Mandal, Development of bioconcrete material using and enrichment culture of novel thermophilic anaerobic bacteria, Indian J. Exp. Biol. 44
(2006) 336–339.
14 T. Manzur et al. / Case Studies in Construction Materials 11 (2019) e00248

[34] T. Manzur, F. Rahman, S. Afroz, R.S. Huq, I.H. Efaz, Potential of a microbiologically induced calcite precipitation process for durability enhancement of
masonry aggregate concrete, J. Mater. Civ. Eng. 29 (5) (2016).
[35] H.M. Jonkers, E. Schlangen, Self-healing of cracked concrete: a bacterial approach, Proceedings of FRAMCOS6: Fracture Mechanics of Concrete and
Concrete Structures (2007).
[36] F. Nosouhian, D. Mostofinejad, H. Hasheminejad, Concrete durability improvement in a sulfate environment using Bacteria, J. Mater. Civ. Eng. 28
(1) (2016).
[37] V.S. Whiffin, Microbial CaCO3 Precipitation for the Production of Biocement. Ph.D. Dissertation, Murdoch Univ., Perth, Australia, 2004.
[38] Z.D. Bundur, M.J. Kirisits, R.D. Ferron, Biomineralized cement-basedmaterials: impact of inoculating vegetative bacterial cells on hydration
andstrength, Cem. Concr. Res. 67 (2014) 237–245.
[39] ASTM, Standard Test Method for Sieve Analysis of Fine and Coarse Aggregates, ASTM C136–06, ASTM International, West Conshohocken, PA, USA, 2006.
[40] Turbidity, Absorptometric Method (1000 FAU), Hach Company/Hach Lange GmbH manual, 04/2013, Edition 8.
[41] ASTM, Standard Test Method for Compressive Strength of Cylindrical Concrete Specimens, ASTM C39 / C39M-15a, ASTM International, West
Conshohocken, PA, USA, 2015.
[42] AASHTO, Standard Method of Test for Predicting Chloride Penetration of Hydraulic Cementby the Rapid Migration Procedure, AASHTO TP 64-03,
American Association of State Highway and Transportation Officials, Washington, DC, USA, 2003.
[43] AASHTO, Standard Method of Test for Electrical Indication of Concrete’s Ability to Resist Chloride Ion Penetration, AASHTO T 277, American Association
of State Highway and Transportation Officials, Washington, DC, USA, 2015.
[44] ASTM, Standard Test Method for Electrical Indication of Concrete’s Ability to Resist Chloride Ion Penetration, ASTM C1202, ASTM International, West
Conshohocken, PA, USA, 2012.
[45] M. Sahmaran, G. Yildirim, R. Noori, E. Ozbay, M. Lachemi, Repeatability and pervasiveness of self-healing in engineered cementitious composites, Aci
Mater. J. 112 (4) (2015) 513.
[46] G. Yildirim, G.H. Aras, Q.S. Banyhussan, M. Şahmaran, M. Lachemi, Estimating the self-healing capability of cementitious composites through non-
destructive electrical-based monitoring, Ndt E Int. 76 (2015) 26–37.
[47] M. Sahmaran, G. Yildirim, E. Ozbay, K. Ahmed, M. Lachemi, Self-healing ability of cementitious composites: effect of addition of pre-soaked expanded
perlite, Mag. Concr. Res. 66 (8) (2014) 409–419.
[48] G. Yıldırım, A.H. Khiavi, S. Yeşilmen, M. Şahmaran, Self-healing performance of aged cementitious composites, Cem. Concr. Compos. 87 (2018) 172–186.
[49] G. Yıldırım, M. Sahmaran, H.U. Ahmed, Influence of hydrated lime addition on the self-healing capability of high-volume fly ash incorporated
cementitious composites, J. Mater. Civ. Eng. 27 (6) (2014).
[50] S. Iffat, A.B. Emon, T. Manzur, S.I. Ahmad, An experiment on durability test (RCPT) of concrete according to ASTM standard method using low-cost
equipment, Adv. Mat. Res. 974 (2014) 335–340.
[51] K.E. Davis, S.J. Joseph, P.H. Janssen, Effects of growth medium, inoculum size, and incubation time on culturability and isolation of soil bacteria, Appl.
Environ. Microbiol. 71 (2005) 826–834.