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A Literature Review of Bio-cement: Microorganisms, Production, Properties,

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Article in ERU Research Journal · October 2023

DOI: 10.21608/erurj.2023.321807

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ERURJ 2023, 2, 4, 554-574
(Review)

A Literature Review of Bio-cement: Microorganisms, Production,

Properties, and Potential Applications.

Osama Ahmed1, Ashraf Abbas2, Ahmed I Mohamed1, 3, Wael Ibrahim2, Sherif Fakhry M.
Abd-Elnaby1
1
Construction Engineering Department, Faculty of Engineering, Egyptian Russian University, Badr City, Cairo
11829, Egypt.
2
Civil Engineering Department, Faculty of Engineering, Helwan University, Al- Matria Branch.
3
Postdoctoral Fellow, Civil and Environmental Engineering Department, University of Missouri, Columbia, 65201,
USA.

*Corresponding author(s): Osama Ahmed, E-mail: [email protected] , Tel: +201098591615

Received 2nd July 2023, Revised 15th October 2023, Accepted 17th October 2023
DOI: 10.21608/ERURJ.2023.321807

1. ABSTRACT
This literature review provides an in-depth analysis of Bio-cement as an Eco-friendly substitute for
conventional cement. Bio-cement is produced using microorganisms that produce calcium carbonate
(CaCO3), which reduces the carbon footprint of cement production. The review covers the different types
of microorganisms used, optimal conditions required for calcium carbonate production, the process of
CaCO3 precipitation, and various test results. Additionally, the paper investigates the potential
applications of Bio-cement in the construction industry, as well as the challenges and limitations
associated with its use. The review highlights several studies on the use of different microorganisms, such
as Sporosarcina pasteurii, Bacillus sp. MCP11, and Bacillus sp. CR2, to create Bio-cement. The
mechanical properties of the final product are also analyzed, including compressive strength, tensile
strength, and flexural strength. The paper provides valuable insights into the potential of Bio-cement as a

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sustainable alternative to conventional cement, as well as the potential limitations and challenges that
need to be overcome to fully realize its benefits. Overall, this literature review is an essential resource for
researchers and individuals interested in the sustainable construction industry. It provides a
comprehensive analysis of the different aspects of Bio-cement, including its production process,
properties, and potential applications. The review serves as a starting point for further research and
development of Bio-cement as a viable and Eco-friendly alternative to conventional cement.
Keywords: Microorganism, CaCo3, Emissions of carbon, MICP, Bio-cement.

2. Introduction

Bio-cement is a sustainable alternative to traditional cement that utilizes microorganisms to produce

calcium carbonate. This process reduces the carbon footprint associated to traditional cement production
and can improve the mechanical properties of the final product. In this paper, we will review the status of
Bio-cement research and development, including the various microorganisms used, the conditions
required for the optimal calcium carbonate production, the CaCo3 Precipitations, and the results from
various tests such as the XRD “X-Ray Diffraction” analysis test, SEM “Scanning electron microscope”
examination test, The FTIR “Fourier-transform infrared spectroscopy” spectrum test, and compressive
strength test, and Biocement potential applications in AEC “Architecture, Engineering, and Construction”
industry. DE Muynck ET. Al. showed that bacterial carbonate precipitation bio-deposition affected the
durability of mortar specimens with varying porosities. Depending on the porosity of the specimens, the
surface deposition of calcium carbonate crystals reduced the water absorption by 85% [14]. Achal and
Pan declared that chromium slag was utilized to produce bricks with MICP application of ureolytic
bacteria Bacillus sp. CS8 for surface treatment. The results of low water absorption of the treated bricks
implied that they had a low permeability [5]. The compressive strength of concrete specimens injected
with B. megaterium was marginally greater than that of B. subtilis-treated concrete samples. Bacillus
sphaericus was used to increase the compressive strength of concrete with a high strength (60 MPa) [4],
according to Shanmuga Priya ET. Al. Bacillus aerius was added to concrete with 10% rice husk ash to
replace some of the cement, increasing its compressive strength from 36 to 40 MPa [3]. According to
Feng et al. sufficient calcium carbonate precipitation on the mortar surface after MICP, treatment by S.
pasteurii could improve the features of recycled fine aggregates (RFAs) of mortars [1]. Iqbal et. al. in
order to produce the bio-OPC cement samples, soil microbial solution with lentil seed powder (a source of
protein) and sugar (a source of carbon) was employed in place of water The bio-bricks' maximum

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compressive strength was found to be 2 MPa [4]. Using the identical kind Chahal, Siddique, and Rajor
found that bacterial and fungal activities such as photosynthesis, ammonification, denitrification, sulphate
reduction, and anaerobic sulphide oxidation could cause calcium carbonate to precipitate extracellularly
[8]. Zaghloul, Ibrahim, and El-Badan concluded Bio cement provides an appropriate replacement for
building projects to reduce the negative environmental effects of cement manufacturing [21].

3. Microorganisms used for Bio-cementation process

Table 1. Types of microorganisms used by Researchers in this Literature review paper:

Authors Microorganism
Heath, Leadbeater, and Callow (1995) Mixture of algae and cyanobacteria collected
from Ely Marina in Cambridgeshire, UK
Rivadeneyra et al. (1996) Deleya halophila, CCM3662 strain
Ramachandran et al. (2001) S. pasteurii
Mortensen et al. (2011) Sporosarcina pasteurii (ATCC 11859)
Santomauro et al. (2012) Scenedesmus obliquus (algae)
Al-Thawadi, Cord-Ruwisch, and Info Bacillus sp. MCP11 (DSM 23526)
(2012)
Stabnikov et al. (2013) S. pasteurii, especially S. pasteurii ATCC
11859
Achal and Pan (2014) Bacillus sp. CR2.
Seifan, Samani, and Berenjian (2016) B. sphaericus and Sporosarcina pasteurii
Xu et al. (2017) Micro-bacterium sp. strain GM-1.
Hait et al. (2018) Klebsiella pneumoniae
Xiao et al. (2021) Sporosarcina pasteurii (DSM33)
Esmail et al. (2022) Bacillus megaterium
Ali, Mukhtar, and Dufossé (2023) Lysinibacillus spp. strain YL

4. Literature review of Biocement techniques and measured variable

Biocement, also known as microbial cement, is a sustainable alternative to traditional cement that has
gained attention in recent years due to its potential environmental benefits. In this literature review, we
will explore the current state of knowledge on Biocement, including its production, properties, and

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potential applications. The paper will also examine the challenges and limitations associated with
Biocement, as well as future research directions. Heath, Leadbeater, and Callow A mixture of algae and
cyanobacteria was collected from the Ely Marina in Cambridge-shire, UK (OS Land Ranger 143, TL 546
798) and cultivated in calcified bio-films. Additionally, nine species of cyanobacteria and algae were
extracted and identified using multiple keys in accordance with the Fritsch collection at the Freshwater
Ecology Institute, Ambleside, UK. There are seven members, one of which is a member of the
Diatomphyta, viz. Members of the genus Navicula and Cyanophyta viz. Synechococcus sp. More than
one medium was used as follows: Isolate single cells using a pulled 1 mm glass tube inserted in a micro-
manipulator. Cells that were isolated were transferred to BBM “Bold's Basal Medium” agar plates, twice-
−2 −1
washed in sterile media, and then incubated at 25°C with continuous illumination (50 A mol m sl ).
Colonies were moved to liquid BBM, where they were frequently subcultured. Additionally, silica was
added to the medium at the final concentration of 200 mg L-1 Na2SiO3.5H2O, or 37.8 mg L-1 Si, for the
separation and development of diatom cultures. And, The calcified medium that provides favorable
conditions for precipitation contains 400 mg L-1 Na2CO3, 400 mg L-1 Na2CO3, 100 mg L-1 MgSO4 2H2O,
13.5 mg L-1 KH2PO4, 25 mg L-1 KNO3, 0.34 mg L-1 1 NaNO2, 2.5 mg L-1 H3BO3, 1.0 mg L-1 (NH4)6
Mo7O24 .4H2O, 1.5 mg L-1 MnC12.4H2O, 25 mg L-1 Na2SiO3 .5H20, 2.0 mg L-1 1.0 mg L-1 of FeCl3 with
EDTA (Na salt). The solution was filter sterilized and the pH “Potential of hydrogen” was changed to be
pH 8.5. The medium is 1195 ps cm-1 in conductivity, 5.79 meq L-1 in alkalinity, and 1.34 in saturation
index (SI) in relation to CaCO3. The sample was made in the following way. To achieve a final
chlorophyll awareness of 0.4 g mL-1, axenic cultures for each alga from the early desk-bound portion were
grown in 250 ml conical flasks with 100 ml of calcification media. The flasks were placed on a gyratory
shaker and incubated at 25°C with constant light (50 mol m-2 s-1). Also created were control flasks with
sterile calcification media. Chlorococcum sp. was used as a further control, with one group of flasks
containing warm-killed algae (1000°C, 2 h), and the other group putting live algae in the dark. Three
copies of each cure were made. For pH (Gallenkamp portable aggregate electrode), soluble calcium
(titrated against 1 mm EDTA “Ethylenediaminetetraacetic” using glyoxal-bis-(2-hydroxy anil) as a
hallmark), and chlorophyll measurements, every two days, a 5 mL sample was obtained from each flask.
Additionally, the main calcification assay utilizing the green unicellular alga Chlorococcum sp. was used
for inhibitor research. The inhibitors had been delivered to the Chlorococcum lifestyle 90 h after sub-
culturing at 10 mg L-1 until in any other case stated. Control flasks had been inhibitor unfastened, each
cure was repeated three times and pH was determined, soluble calcium and chlorophyll. In which

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inhibitors had been brought at several concentrations a good way to confirm the minimal awareness
powerful at regulating precipitation and the dosage at which 100% inhibition changed until achieved.
Additionally, inhibitor experiments using the Stigeoclonium variable were completed. The Olympus BH-
2 microscope and transmitted light polarizing filters were used., samples have been checked for CaCO3
precipitation during each test. Black and white Kodak TMAX 400 film has been used to shoot pictures.
The sampling's outcomes were as follows: In all the flasks of calcification medium with no algae,
precipitation was no longer seen, and the levels of soluble calcium and pH remained at initial levels.
When cultured in calcification media, every type of algae extracted from calcified biofilms deposition
CaCO3. Tannic acid, ((NaPO3)),(Na4P2O7), Induction times PAA and a 92:8 mole % mixture of polyprotic
acid and alanine (PAL) extended the experiments, and 50 hours after these compounds were given to the
flasks, there were no signs of precipitation. Additionally, most of these flasks had deposited various
quantities of CaCO3 after 100 hours of use. The other substances may also have delayed precipitation, but
this test was unable to detect it since there was a period between samplings during which conditions were
no longer being watched. Additionally, polyaspartic acid MW 2000 (PA2000), tannic acid, PAL, PAA,
(KH2PO4), MDPA and PA250000 polyacrylic acid decreased the rate at which soluble calcium was
eliminated from the medium during precipitation. In comparison to the controls, the rate at which the
removal of soluble calcium from the medium was enhanced in the presence of (NaPO3)6 and Na4P2O7.
The pH of the flasks containing inhibitor, on the other hand, rose by at least 0.69 units above the pH at
which precipitation initially appeared. They were successful in precipitating calcium carbonate. When
(NaPO3)6 and Na4P2O7 were introduced to the flasks at 100 mg L-1, the CaCO3 precipitation was
completely inhibited. Furthermore, they observed that these algae are susceptible to high pH values, with
death occurring when the pH reached 10.4. HEDP was the most efficient inhibitor of crystal nucleation.
Chlorophyll was used to quantify growth, and the growth rates were determined by calculating the
gradient of the slope that resulted from a plot of log chlorophyll vs. time. The slope of a plot of the
concentration of soluble calcium against time was used to calculate the rates of soluble calcium
elimination. Divide the rate of calcium elimination by the rate of growth to get, the ability of an alga to
precipitate CaCO3 was determined. The slope of a plot of the concentration of soluble calcium against
time was used to calculate the rates of soluble calcium elimination. By dividing the rate of calcium
removal by the rate of growth, the ability of an alga to precipitate CaCO3 was determined. The research
did not stop to this extent [11]. Rivadeneyra et. al. Used Deleya halophila, CCM3662 strain. With
Medium: The bacteria were cultured on MH liquid medium that had 0.4% calcium acetate added to it. The

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pH was then altered to 7.2 using 1 M KOH. The MH medium was modified to contain 1% (wt/vol) yeast
extract (Difco), 0.5% proteose peptone No. 3, and 1% glucose in addition to a well-balanced combination
of sea salts with a 7.5% (wt/vol) final salt concentration. In order to observe the crystallisation process, D.
Halophila was also planted in liquid media in widemouthed vials that had previously been filled with
vertically positioned glass coverslips. The bottles were incubated at 32°C, and after 120 hours, the
coverslips were taken off for examination every 12 hours. The slides were preserved using CaCl2 in a
desiccator. Additionally, a liquid medium was used to grow microorganisms for 25 days to produce
enough crystals to determine the mineralogical features. The crystals were collected, cleaned with
distilled water, and allowed to dry naturally at 37 °C. The final outcomes were as follows: The major
mineral in the crystals produced by D. halophila is aragonite (CaCO3). This species' bio minerals emerged
after 25 days of incubation, and magnesium levels were minuscule throughout the biolith growth process.
Unlike other polymorphic forms of calcium carbonate, like calcite, very little magnesium substitutes for
calcium in the crystalline network of aragonite. The sample was produced using a Link QX-200
equipment coupled to a Zeiss DSM-950 scanning electron microscope, and after completion, X-ray micro
analyzer (EDX) tests were to be conducted on the sample. At 20 kV, 100 s of existence time, a spot size
of approximately 200 nm, and a 45° tilt attitude, the samples were evaluated using pin-factor analysis.
Dolomite (Ca 0.5-Mg 0.5-CO3) and high-purity calcite (CaCO3) have been used as controls. These items
have been coated with carbon and set up on a graphite pattern plate with carbon glue. And, the result of
the test is as follows: calcium became the major alkaline-earth steel, and was followed by means of
negligible quantities of magnesium. Since Halophila in particular mineralizes calcium, biomineralization
via this species is fundamentally a calcification process. Despite the fact that the living medium included
more of the latter metal (1094 ppm calcium compared to 2717 ppm magnesium), the biolithic created by
this species had significantly more calcium than magnesium. After that, a light microscope was done and
the result Within 48 h after seeding, Bio minerals were generated by D. Halophila in spherical forms. The
investigation didn't end there; SEM analysis was also carried out, with the following findings:
Morphological studies with SEM of the levels of crystal formation revealed that the first indications of
biomineralization appeared after around 24 hours of incubation. However, with time, both the percentage
of biolithic and the percentage of bio-minerals that reflect the very final ranges of the precipitation series
increased. After 25 days of development, the XRD examination was completed, and the results revealed
that the mineral section was mostly composed of aragonite (polymorphous calcium carbonate), with trace
levels of magnesium calcite. It has been accomplished to reach: Due to the fact that Deleya halophila is a

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Gram-negative encapsulated bacterium, the buildup of metal ions (Ca+2, with a lesser percentage of Mg+2)
seen in our micro-analytical data most likely occurs within the cell wall and capsule. The adsorption of
Ca+2 and Mg+2 cations at the cell surface by bacteria can operate as the center for the precipitation of
carbonates, according to several authors. The process of carbonate precipitation involves the adsorption of
Ca+2 cations (and of a minor quantity of Mg+2 cations), which is followed by a local explosion in their
concentration. Along with the emission of CO2 caused by the decomposition of organic materials, The
local supersaturation and precipitation of calcium carbonate, primarily aragonite, may also result from the
enhanced focus [15]. Research began in the 21st century; similarly, Ramachandran et al. demonstrated that
embedding S. pasteurii cells into the cement matrix increased the compressive power of cement mortar.
The calcium carbonate crystals produced by Lysinibacillus spp. strain YL under identical circumstances
have not been subjected to X-ray diffraction investigation [22]. Katsuyama et al. prepare the sample as
follows: waste cement sample utilized in this investigation (Tokyo, Japan). The sample is made up of tiny
particles that were separated and pulverized as byproducts of a waste recycling factory for concrete. By
using a light Scattering technique, the diameters of particles of the waste cement were examined and
found to be spread throughout a range of 10-200 mm, peaking at roughly 25-40 m (area based) and 80 m
(volume based). According to elemental analysis, calcium made up roughly 27.3% of the weight fraction.
Based on thermogravimetric measurements made with the aid of a differential thermal analyzer, it was
discovered that around 11% of the calcium had already carbonated (TGD-9600, Ulvac, Tokyo, Japan).
Additionally, the extraction of calcium tests was carried out in a high-pressure stirring tank vessel reactor.
The interior volume was 500 mL, and the container was built of the nickel-based alloy Hastelloy. With
air-conditioning conditions, a known amount of waste cement particles and a set amount of ultrahigh-
purity water were introduced into the reactor. The reactor was continually supplied with gaseous CO2.
The reaction temperature was precisely maintained while the reactor was immersed in a constant
temperature bath with an accuracy of ±1 K. Additionally, the material in the reactor was stirred using a
two-wing paddle-type fin, with the speed of the churning being adjustable between 0-1000 rpm.
Throughout the extraction trials, small samples of the reactor's content were taken through a sintered
metal filter with a mesh size of 5 µm at predetermined intervals. Inductively coupled plasma-atomic
emission spectrometry was used to determine the calcium concentration of the filtered solution. As
calcium carbonate may be the reason of the apparent extraction rate indicated above, consider the
mismatch between the rate at which calcium ions are extracted from the waste cement and the rate at
which they are precipitated with carbonate ions. The extraction rate will be significantly larger than the

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precipitation rate, especially early in the reaction, when there is a lot of waste cement available for the
extraction reaction and the C/W ratio is higher. This explains the hypersaturation that was seen during the
first stages of the extraction process in the instances with higher C/W ratios. In the end, he predicted that
as the extraction reaction advanced and the concentration of calcium ions in the aqueous phase grew, so
would the amount of waste cement available for the extraction process. And, when figuring up the
required C/W ratio, these trade-offs should be taken into consideration. Additionally, all the tests
involving the precipitation process employed the aqueous solution created during the extraction
procedure. After passing through a sintered metal filter with a mesh size of 5 m, the solution generated by
the extraction reaction under the prescribed extraction conditions was passed to the precipitation reactor.
Additionally, the precipitation reactor is a 300 mL reinforced glass jar. The precipitation process is
triggered by either increasing temperature or decreasing CO2 pressure. ICP-AES was used to determine
the calcium concentration after a very little amount of the solution was filtered through a mesh size of 5
m. Following that, to measure the rate of calcium carbonate precipitation, A P-4010 (Hitachi) was used to
quantify the concentration of calcium ions in the solution at regular intervals. The precipitation process
was allowed to proceed for a certain period, and then the entire reactor contents were withdrawn and
immediately filtered. Additionally using a differential thermal analyzer and the thermos gravimetric
technique (DTG-60H, Shimadzu, Kyoto, Japan), the chemical makeup of the particles that remained on
the filter was studied. According to the thermal gravimetric measurement, the precipitated CaCO3
concentration was greater than 98%. It should be emphasized considering these findings that the
formation of the required high-purity CaCO3 is contingent on the existence of seed crystals. Calcium
carbonate has an 80% purity without seed crystals. As a result, during the precipitation experiments, the
rate of calcium carbonate precipitation is almost precisely similar to the decrease in calcium concentration
in the solution. After that, he kept looking into the impact of CO2 changes in the concentration of calcium
ions under various partial CO2 gas pressures. The prescribed variables are a temperature of 323 K, stirring
speed of 900 rpm, and 1.0 g of residual cement. The dotted lines on Figure 1 represent the saturated
calcium concentrations for the selected extraction conditions. Effect of CO2 partial pressure on the
calcium ion concentration of the solution over time. The starting solution volume was 200 mL, the
precipitation temperature was 303 K, the stirring speed was 500 rpm, and the CaCO3 seed crystal weight
was 0.05 g. The Figure 1 shows dotted, dashed, and solid lines, respectively, represents the estimated
saturation concentration of Ca+2 under these conditions and is based on thermodynamic principles.

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Figure 1. Temperature of precipitation influences the time course of calcium ion concentration in solution. Initial
solution volume = 200 mL, CO2 partial pressure = 0.2 MPa, stirring rate = 500 rpm, CaCO3 seed crystal quantity =
0.05 g. The dotted, dashed, and solid lines on the graph represent the Ca2+ saturation concentrations determined
using thermodynamic principles [12].

Finally in terms of cost and energy utilization, the proposed method for manufacturing calcium carbonate
from waste cement using compressed CO2 has substantial promise for flue gas desulfurization [12]. In a
study by Mortensen et al. Biocement was made with the aid of Sporosarcina pasteurii. A urea-calcium-
based cementation medium was used to culture the bacterium, which was then incubated for around 40
hours before being extracted. The researchers were able to delay the precipitation of calcium carbonate
and produce a more uniform dispersion of cementation by raising the ammonium chloride content in the
cementation medium. The tests were conducted using Scenedesmus obliquus (strain 276-2) [13]. A live
organism Santomauro et al., It came from the Gottingen SAG cultural collection. In addition, Zn+2,
Na2EDTA, KH2PO4, (NH4)Mo7O24, and MnCl2 were eliminated from the modified calcification medium
on which the algae were grown in order to minimize chelate formation, and FeCl3 was reduced to 0.01
mg/l. The solution was saturated for Ca2+. ZnSO4.7 H2O, which contains zinc, was added to the medium
at three different concentrations (0, 3.27, and 6.53 mg Zn+2/l).Additionally, all stock solutions — aside
from Na2CO3 were combined, and sterilized in a Systec V-75 autoclave, and their pH levels were elevated
to 6.3. The algae were washed in decriminalized water (Millipore), counted with a hem cytometer
(Marienfeld, Lauda-Königshofen), and then added to the medium (around 4.0 x 108 cells/l) to reach the
same cell content in all cultures. The culture was kept alive in an inform HT Multitron II rotary shaker at
a constant 26 C under Gro-Lux 15W, 3500 lx fluorescent illumination. Due to photosynthesis, the pH rose
over 9 over the course of many days. Daily pH adjustments were made using 0.1 M NaOH in both the
organic, algae-containing solution and the algae-free solution. The 72-hour experiment was conducted.

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All cultures thereafter adopted the lifestyle that was made apparent by their green colour. The results of
the samples gathered were also reported. classification of poly-morphs and their crystallisation. The
occurrence of precipitates in solutions containing algae was investigated at the start of the experiment and
at intervals of 2 hours, 4 hours, 6 hours, 24 hours, and 72 hours. There were found to be two different
types of crystals. According to the crystal form, we assume that the calcite-representing rhombohedric
crystals and the aragonite-representing needle-like crystals are both minerals. Scenedesmus obliquus does
not calcify when cultivated in the dark, and the pH only rises to a maximum of 9. Through light-
dependent photosynthesis, the algae actively contribute to the alkalization of the medium in addition to
acting as CaCO3 precipitation nucleation sites. And for the pH, each culture's pH was adjusted to 8.5
before to the commencement of each experiment. The pH rose to 10.8 after 48 hours in the culture
without zinc as a result of the algae's photosynthesis; following that, it practically stayed constant. The pH
change was postponed by zinc, preventing it from rising to the high levels of the zinc-deficient culture.
While the culture containing 3.27 mg Zn+2/l reached pH 10.5 after 48 hours and stayed there, the culture
containing 6.53 mg Zn+2/l only reached pH 10.1 after 72 hours. This pH-shift is related to the amount of
Zn+2 present in the solution. Using a 0.1 M NaOH solution, daily pH changes were performed to the
media devoid of algae in line with the pH readings taken from cultures containing algae, then for SEM
analysis The outcomes were Only aragonite crystals are discovered in the medium, which also contains
significant quantities of zinc and algae. On the other hand, calcite nearly always forms in environments
with high zinc concentrations and low amounts of algae. The amount of aragonite produced is quite little.
The findings of the XRD examination also showed, the presence of algae cells greatly changes the CaCO3
precipitation within the medium as compared to medium without algal cells. Finally, we can state that the
presence of living micro-algae has a considerable impact on the precipitation of calcium carbonate
crystals. As a result of photosynthesis, the pH of the organic media containing the algae increased, and
CaCO3 crystals developed. In organic media, 3.27 mg Zn+2/l or not, calcite and aragonite always form. In
organic settings with 6.53 mg Zn+2/l, even aragonite forms. In contrast, inorganic solutions devoid of zinc
precipitate pure calcite. Both of the inorganic solutions containing zinc had significant calcite
precipitation and a smaller amount of aragonite precipitation [16]. Similarly, in a study of Al-Thawadi,
Cord-Ruwisch, Biocement was created using Bacillus sp. MCP11. The bacterium, which was cultivated in
a medium containing yeast extract, urea, ammonium sulphate, sodium acetate, and calcium chloride, was
isolated from soil and sludge samples. The scientists watched as originally spherical crystals formed,
progressively fragmented, and then finally transformed into rhombohedral crystals. The production of

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calcite and vaterite crystals was shown by X-ray diffraction examination of the calcium carbonate
precipitated by MCP11 under comparable circumstances[8]. Then Stabnikov et al. did a study on the
production of bio-cement using S. pasteurii, notably the ATCC 11859 strain. Pure cultures of UPB were
gathered by the researchers from a variety of climates, including halophilic and alkalophilic strains from
Singaporean and Ukrainian soil. The bacteria were grown in a medium that included trace elements as
well as 82.5 g/l (0.75 M) of calcium chloride and 90 g/l (1.5 M) of urea. Using 1 M HCl, the pH of the
mixture was brought down to 2.0. Ten grams of dirt were used to inoculate the medium, which was then
shaken for six days at 150 rev/min at 30°C. After six batches of treatments with strains VS1 and VUK5,
the researchers found that the unconfined compressive strengths for dry
bio-cemented sand samples were 765 and 845 kPa, respectively. The study demonstrated the potential of
S. pasteurii for usage in building applications by proving its ability to create bio-cement [18]. Also, Achal
and Pan, Bacillus sp. CR2, which was isolated from mine tailing soil in Urumqi, Xinjiang, China, was
used to study the formation of calcium carbonate. Using 16S rRNA gene sequencing, a thorough
molecular investigation was used to identify the bacteria. The bacteria were cultivated in Nutrient broth
supplemented with urea and several calcium sources, such as calcium chloride, calcium oxide, and
calcium acetate, in order to create calcium carbonate. The XRD spectra revealed that Bacillus sp. CR2
primarily formed calcite in all media, with the most calcite peaks being seen in the calcium chloride-
containing solution. In all media, with the exception of the one that used calcium acetate as the calcium
source, aragonite and vaterite peaks were also seen. Calcite, vaterite, and aragonite were among the
mineral poly-morphs that were discovered by SEM analysis. Many of the calcite crystals also included
rod-shaped bacterial cells that were intimately linked to and emerging from the mineral surface. After
growing bacterial cells in various calcium sources for 7 days, the cell pellets were examined using the
FTIR spectra produced by infrared spectroscopic analysis. Bacillus sp. CR2 generated the most calcium
carbonate (2.32 mg) when calcium chloride was used as the calcium source, according to the research,
making it the most favoured calcium source [5]. After that Seifan, Samani, and Berenjian used
Sporosarcina pasteurii and B. sphaericus to aid in the formation of bio-minerals. The biological materials
employed in this investigation were discovered to be mediated by a wide range of microorganisms,
including bacteria, fungus, protists, and plants. Numerous objects, including shells, bones, teeth, and even
limestone caverns, contain these bio-minerals. As the main calcium supply for the experiment, calcium
carbonate, or CaCO3-H2O, was employed [17]. Similarly, in a study Xu et al., strain GM-1, a Micro-
bacterium sp. isolated from active sludge, was used to induce calcium carbonate precipitation through

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urea hydrolysis. The microorganism was incubated aerobically at 30°C in a unease-selective medium.
Using the streak plate technique, the cells were inoculated on urease selective and control plates, and
color change was seen after 2 days of incubation. Calcite was found to be the predominant calcium
carbonate form in the XRD study of the white precipitate produced by the bacterium groups. According to
the study, strain GM-1 may be useful in environmental bio-remediation and bio-recovery [20]. Also in
Hait et al. completed a study employing Klebsiella pneumoniae to generate bio-cement. For the purpose
of isolating the organisms that produce bio-cement, the researchers took soil, seawater, and sewage
samples. The researchers used titrimetric analysis to check for the presence of Ca2+ ions in hard water
systems, such as saltwater and sewage water, in order to produce bio-cement. The calcium source
employed was the original seawater's Ca2+ concentration. The presence of calcium carbonate was further
confirmed by the XRD examination of the synthetic bio-cement, which is compatible with earlier
research. The signature peaks at 2θ = 29.8°(104) and 43.7 (202) clearly showed the presence of CaCO3.
However, it is also clear from the pattern that calcite is not created in its pure form; peaks at 2 =16.7,
21.7, and 32.4 reflect different crystal structures of aragonite and dolomite, both of which are found in the
bio-cement made by Klebsiella pneumoniae as shown in Figure 2.

Figure 2. XRD analysis of white precipitate, obtained from isolate Klebsiella pneumoniae, [10]

The test organism's production of bio-cement was determined to be 0.46 g/1000ml in saltwater. The fact
that there was less Ca2+ in the filtrate proved that Ca2+ was used for CaCO3 precipitation [10]. Similar to
this, Sporosarcina pasteurii (DSM33) was employed in a recent work by Xiao et al. to investigate calcium
carbonate precipitation. The bacteria were obtained from Leibniz Institute DSMZ and was raised in a
liquid medium comprising 0.13 M Tris buffer, 20 g of yeast extract, and 8.1 g of NH4Cl. The bacteria
culture become murky after 20–24 hours of incubation at 30 °C, signifying the development of the

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bacterium. For the XRD analysis, fragments were crushed and sieved through a 75 µm mesh. The analysis
revealed the formation of calcium carbonate crystals, and the XRD scan was conducted using
Cu Ka radiation (40 kV, 30 mA) with a scanning rate of 0.017° 2θ/step from 5° to 80° 2θ
as shown in Figure 3 [19].

Figure 3. XRD patterns of RMC powder, control RMC paste (mix l), nutritious RMC paste (mix 7), and bio-RMC
paste (mix 8) at 28 days, [19].

After that Esmail et al. did research on using Bacillus megaterium to make bacterial concrete. Using
MALDI-TOF Biotyper, the bacterium was isolated from alkaline soil samples collected in Wadi EL-
Nitron, Behera governorate, Egypt. The culture was grown in aerobic incubation in 2L Erlenmeyer flasks
using a rotary shaking incubator at 150 rpm for 7 days at 30°C using autoclave Nutrient broth (NB) with a
pH of 7.5. Two stages of work were carried out with Bacillus megaterium for their investigation. In the
first step, calcium lactate, calcium formate, and calcium acetate were introduced as nutrition to bacteria at
0.25%, 0.125%, and 0.5% of cement weight, respectively, to concrete mixes in two ratios (0.5% and
0.25% of cement weight). Bacillus megaterium was introduced to concrete mixes in the second stage in
two ratios (0.5% and 0.25% of cement weight), with calcium lactate provided as nourishment to bacteria
in the ratios of 0.125% and 0.250% of cement weight. In concrete mixes containing 1% cement,
superplasticizer was utilized. According to SEM imaging, the addition of bacterial concrete specimens'
nutrients caused calcite crystals to develop in a variety of forms. According to earlier research, bacterial
concrete has a better compressive strength than regular concrete. In comparison to normal concrete, the
inclusion of Bacillus megaterium, Bacillus subtilis, and their consortia enhanced compressive strength by

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22.5%, 14.3%, and 15.8%, respectively. Additionally, when fly ash concentrations were 10%, 20%, and
40%, respectively, bacterial cells increased the compressive strength of mortar by 19%, 14%, and 10% in
comparison to control specimens. In comparison to the standard concrete strength of 44 MPa, the
employment of Bacillus subtilis at a concentration of 108 cells/ml resulted in a compressive strength of 52
MPa, while Bacillus megaterium at a concentration of 105 cells/ml produced a compressive strength of 57
MPa. Due to the calcification process, the strength of the highest grade of bacterial concrete (50 MPa)
increased by 24% compared to the lowest grade (30 MPa) by 12.8%. The presence of bacteria improves
the compressive strength of silica fume (SF) concrete at all ages, with the greatest strength of bacterial
concrete being recorded at 56 days with 10% SF and being around 12% higher than that of the concrete
with the same silica fume replacement. The deposition of calcite in the pores, followed by pore reduction,
compact micro-structure formed, rendered concrete thick and so improving the strength, may be the cause
of the increase in concrete strength with bacteria. The negatively charged groups on the surface of
bacterial cells, which attract divalent ions like Ca2+ and Mg2+, serve as a location for nucleation. Urea is
hydrolyzed by the urease enzyme into ammonia and carbonate, which then react with Ca2+ to generate
calcium calcite precipitate on the bacterial surface. The concrete mix surrounding the pores and micro-
cracks is better packed and compacted as a result of the calcite, giving the specimens a significantly
greater strength than controlled concrete examples. Calcium carbonate also accelerates hydrolyzation and
serves as a catalyst for cement hydration, boosting the concrete's compressive strength [9]. And finally,
Ali, Mukhtar, and Dufossé used the Lysinibacillus spp. strain YL to bio-precipitate calcium carbonate
crystals. The microorganism was isolated from tropical beach sand and raised in a medium with calcium
sources such as acetic acid or calcium acetate. An efficient and cost-effective method for bio-
precipitation, according to the study, would involve employing a low concentration of Ca2+ as low as 30
mM as shown in Figure 4. also showed that non-ureolytic Bacillus cohnii may be used to perform
microbially induced calcium carbonate precipitation (MICCP), which might boost concrete's compressive
strength by 49% [6].

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Figure 4. Schematic of MICCP crack healing - cracks form in bio-cement with spores and Ca+2. Water and air
seepage release encapsulated spores and Ca+2 [6]

5. The effects of different parameters on the biocementation process

Firstly, the choice and concentration of nutrients introduced into concrete mixes are of paramount
importance. In the study, calcium lactate, calcium formate, and calcium acetate were employed as
nutrients at different levels, both in absolute concentration and as percentages of cement weight. The
concentration of nutrients directly influenced the formation of calcite crystals within the concrete matrix.
The results revealed that the choice and concentration of these nutrients significantly affected the size,
morphology, and distribution of calcite crystals. This underscores the potential for tailoring the
biocementation process to achieve specific material properties by optimizing nutrient composition.
Furthermore, the concentration of the introduced bacterial strain is a key parameter. Bacillus megaterium,
at different concentrations, showed varying effects on the compressive strength of the resulting concrete.
The study demonstrated a remarkable enhancement in compressive strength, ranging from 14.3% to
22.5% compared to conventional concrete. At a concentration of 10^5 cells/ml, Bacillus megaterium
achieved a notable compressive strength of 57 MPa. This emphasizes that the concentration of the
bacterial strain can be fine-tuned to attain desired concrete properties, presenting a strategy to engineer
concrete materials with superior mechanical characteristics. Additionally, the research delves into the
impact of fly ash concentrations on the compressive strength of concrete. The incorporation of fly ash
further improved the compressive strength of mortar specimens, with enhancements of up to 19%
compared to control specimens. This parameter demonstrates the potential for using supplementary

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materials to enhance the biocementation process, thereby offering eco-friendly alternatives in concrete
production while optimizing mechanical performance.

6. Different strategies used for the biocementation process

One notable strategy involves the use of Bacillus sp. strains, such as Bacillus megaterium and Bacillus
subtilis, which have exhibited exceptional abilities to induce calcium carbonate precipitation. These
bacterial species are capable of altering the material properties of concrete through their metabolic
activities. By introducing Bacillus sp. strains to concrete mixes at varying concentrations, the
biocementation process can be fine-tuned to enhance compressive strength significantly. The
incorporation of these bacterial strains into concrete mixes has resulted in concrete materials with
compressive strengths ranging from 30 MPa to 50 MPa. This strategy not only offers a sustainable
approach to concrete production but also provides an opportunity to engineer materials with superior
mechanical performance. Another strategy employs microorganisms like Sporosarcina pasteurii,
Klebsiella pneumoniae, and Lysinibacillus spp. to initiate the precipitation of calcium carbonate in
concrete matrices. These microorganisms are isolated from various sources, including soil, sewage, and
tropical beach sand. Each microorganism's unique characteristics and metabolic processes influence the
resulting material properties. The use of different microorganisms demonstrates the adaptability and
versatility of the biocementation process. In addition to microbial species, the choice of nutritional
composition and supplementary materials plays a pivotal role in optimizing the biocementation process.
Nutrient selection, such as calcium sources like calcium lactate, calcium formate, and calcium acetate,
directly impacts the formation and distribution of calcite crystals within the concrete matrix. This offers
the possibility of tailoring material properties to meet specific requirements in construction applications.
Furthermore, the inclusion of supplementary materials like fly ash showcases the strategy of incorporating
eco-friendly materials into the biocementation process. Fly ash concentrations, when combined with
microbial activity, further enhance the compressive strength of concrete, presenting sustainable
alternatives for concrete production with improved mechanical properties.

7. Utilization of microbial-induced calcite precipitation for various applications

Microbial-induced calcite precipitation (MICCP) has emerged as a versatile and promising technology
with a wide range of applications across various fields. This biologically mediated process is being
extensively explored for its potential to revolutionize the construction industry, environmental

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remediation, and materials science. A notable application of MICCP within the construction industry is
the enhancement of concrete properties. Research endeavors involving bacterial strains like Bacillus
megaterium and Bacillus subtilis have demonstrated impressive results. The introduction of these bacteria
into concrete mixes led to substantial improvements in compressive strength, ranging from 14.3% to
22.5% when compared to traditional concrete formulations. The use of fly ash in combination with
bacterial cells further boosted the compressive strength of mortar specimens. Bacillus subtilis at a
concentration of 10^8 cells/ml resulted in a compressive strength of 52 MPa, while Bacillus megaterium
at a concentration of 10^5 cells/ml produced a remarkable compressive strength of 57 MPa. The highest-
grade bacterial concrete exhibited a 24% increase in strength over the lowest grade, highlighting the
efficacy of the calcification process in enhancing concrete strength. The benefits of MICCP in the
construction sector extend to silica fume (SF) concrete, with the greatest strength observed at 56 days
when 10% SF was incorporated. The improved compressive strength recorded at this juncture was
approximately 12% higher than that of the control concrete with the same silica fume replacement, further
underlining the potential of bacterial influence on concrete properties. Also, MICCP has garnered
attention for its eco-friendly application in environmental remediation. Notably, the biocementation
process facilitates the stabilization and immobilization of contaminants in soils. The precipitation of
calcium carbonate in soil matrices can enhance soil cohesion and reduce permeability, thus reducing the
mobility of harmful substances. This method provides an innovative approach for the treatment of
contaminated sites, including those with heavy metal or radionuclide pollution. MICCP can significantly
decrease the mobility of these contaminants, contributing to the sustainable remediation of soil and
groundwater. Notably, in a study by Esmail et al., Bacillus megaterium successfully facilitated bacterial
concrete production. The ability of this bacterium to enhance compressive strength makes it a promising
candidate for creating bio-cement with environmental applications, including soil stabilization. In
materials science, MICCP offers unique prospects for the production of biocement and biominerals. The
research conducted by Xiao et al. showcased the feasibility of producing calcium carbonate crystals using
Sporosarcina pasteurii. These crystals were formed in controlled environments, and their utilization for
various applications was explored. The biomineralization process can be engineered to produce specific
mineral phases and morphologies, opening doors for tailored materials with a multitude of applications in
the fields of medicine, dentistry, and materials engineering. The cost-effectiveness of MICCP for
biomineral production is exemplified by the use of low concentrations of Ca^2+, as low as 30 mM. The
study suggests that this innovative approach may revolutionize the production of biominerals for diverse

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industrial and medical applications. MICCP is a multifaceted and promising field with the potential to
address pressing challenges in construction, environmental protection, and materials science. The
quantifiable benefits, including enhanced concrete strength, reduced soil permeability, and cost-effective
biomineral production, underscore the significance of MICCP as a sustainable and innovative technology
with broad-ranging applications. As research in this area continues to advance, the full extent of its
potential and practical implications is yet to be fully realized, making it an exciting area of exploration in
scientific and engineering disciplines.

8. Conclusion
In conclusion, Biocement is an Eco-friendly substitution for traditional cement that utilizes
microorganisms to produce calcium carbonate and reduce the carbon footprint of cement production. The
literature review explored the various microorganisms, also the optimal conditions for calcium carbonate
production, CaCO3 precipitations, and potential applications of Biocement in the construction industry.
The current study also examined the challenges and limitations associated with Biocement and
highlighted the need for further research. The reviewed studies showed that Biocement has the potential to
improve the mechanical properties of the final product and using this technique could decrease the
environmental impact of cement production. Table [1] provides an overview of the researchers who have
conducted studies in this area, along with the specific type of bacteria or algae that was used. This
literature review study is a helpful for researchers and individuals interested in the sustainable
construction sector.

• Acknowledgment
The authors are thankful for the kind support from the Egyptian Russian University to carry
out the research work.
• Conflict of Interest
A declaration of conflict of interest.

9. References

[1] Feng, Zhangyao & Zhao, Yuxi & Zeng, Weilai & Lu, Zhenmei & Shah, Surendra. (2020). Using
microbial carbonate precipitation to improve the properties of recycled fine aggregate and mortar.
Construction and Building Materials. 230. 116949. 10.1016/j.conbuildmat.2019.116949.

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 ERURJ 2023, 2, 4, 554-574

[2] Sumit Joshi, Shweta Goyal, Abhijit Mukherjee, M Sudhakara Reddy, Microbial healing of cracks in
concrete: a review, Journal of Industrial Microbiology and Biotechnology, Volume 44, Issue 11, 1
November 2017, Pages 1511–1525, https://doi.org/10.1007/s10295-017-1978-0

[3] Thangaraj, Shanmuga Priya & Ramesh, N. & Agarwal, Ankit & Bhusnur, Shreya & Chaudhary,
Kamal. (2019). Strength and durability characteristics of concrete made by micronized biomass silica
and Bacteria-Bacillus sphaericus. Construction and Building Materials. 226. 827-838.
10.1016/j.conbuildmat.2019.07.172

[4] Iqbal, Dawood Muhammad, Leong Sing Wong, and Sih Ying Kong. 2021. "Bio-Cementation in
Construction Materials: A Review" Materials 14, no. 9: 2175. https://doi.org/10.3390/ma14092175

[5] Achal, Varenyam, and Xiangliang Pan. 2014. “Influence of Calcium Sources on Microbially Induced
Calcium Carbonate Precipitation by Bacillus Sp. CR2.” Applied Biochemistry and Biotechnology 173
(1): 307–17. https://doi.org/10.1007/s12010-014-0842-1.

[6] Ali, M. F., H. Mukhtar, and L. Dufossé. 2023. “Microbial Calcite Induction: A Magic That Fortifies
and Heals Concrete.” International Journal of Environmental Science and Technology. Institute for
Ionics. https://doi.org/10.1007/s13762-022-03941-2.

[7] Al-Thawadi, Salwa, Ralf Cord-Ruwisch. 2012. “Calcium Carbonate Crystals Formation by Ureolytic
Bacteria Isolated from Australian Soil and Sludge Biosynthesis of Nanoparticles View Project
Investigating the Role of Electron Mediators in Microbial Extracellular Electron Transfer (EET)
View Project Calcium Carbonate Crystals Formation by Ureolytic Bacteria Isolated from Australian
Soil and Sludge.” Journal of Advanced Science and Engineering Research 2: 12–26.
https://www.researchgate.net/publication/230603445.

[8] Chahal, Navneet, Rafat Siddique, and Anita Rajor. 2012. “Influence of Bacteria on the Compressive
Strength, Water Absorption and Rapid Chloride Permeability of Concrete Incorporating Silica
Fume.” Construction and Building Materials 37 (December): 645–51.
https://doi.org/10.1016/j.conbuildmat.2012.07.029.

[9] Esmail, Randa F, Rateb N Abbas, Noha M Sorour, and Amal A Nasser. 2022. “Effect of Bacillus
Megaterium Bacteria and Different Calcium Sources on Strength and Permeation Properties of
Concrete.” Civil Eng 45 (3): 401–12.

[10] Hait, Supriya, Gauri Chemburkar, Divya Murlidhar, Ulrica Almeida, Ruth Fernandez, Vivien
Amonkar, and Pampi Chakraborty. 2018. “Strengthening Water Retention Capacity of Marine Soft
Clay Using Bio-Cement.” Applied Ecology and Environmental Sciences 6 (3): 93–96.
https://doi.org/10.12691/aees-6-3-4.

572
 ERURJ 2023, 2, 4, 554-574

[11] Heath, Carolyn R, Barry C S Leadbeater, and Maureen E Callow. 1995. “Effect of Inhibitors on
Calcium Carbonate Deposition Mediated by Freshwater Algae.” Journal of Applied Phycology. Vol.
7. Kluwer Academic Publishers.

[12] Katsuyama, Yasuro, Akihiro Yamasaki, Atsushi Iizuka, Minoru Fujii, Kazukiyo Kumagai, and
Yukio Yanagisawa. 2005. “Development of a Process for Producing High-Purity Calcium Carbonate
(CaCO3) from Waste Cement Using Pressurized CO2.” Environmental Progress 24 (2): 162–70.
https://doi.org/10.1002/ep.10080.

[13] Mortensen, B. M., M. J. Haber, J. T. Dejong, L. F. Caslake, and D. C. Nelson. 2011. “Effects of
Environmental Factors on Microbial Induced Calcium Carbonate Precipitation.” Journal of Applied
Microbiology 111 (2): 338–49. https://doi.org/10.1111/j.1365-2672.2011.05065.x.

[14] Muynck, Willem De, Dieter Debrouwer, Nele De Belie, and Willy Verstraete. 2008. “Bacterial
Carbonate Precipitation Improves the Durability of Cementitious Materials.” Cement and Concrete
Research 38 (7): 1005–14. https://doi.org/10.1016/j.cemconres.2008.03.005.

[15] Rivadeneyra, M A, A Ramos-Cormenzana, G Delgado, and R Delgado. 1996. “Process of Carbonate

Precipitation by Deleya Halophila.” An International Journal. Vol. 32. Springer-Verlag New York
Inc.

[16] Santomauro, Giulia, Johannes Baier, Wanjing Huang, Stefan Pezold, and Joachim Bill. 2012.
“Formation of Calcium Carbonate Polymorphs Induced by Living Microalgae.” Journal of
Biomaterials and Nanobiotechnology 03 (04): 413–20 https://doi.org/10.4236/jbnb.2012.34041.

[17] Seifan, Mostafa, Ali Khajeh Samani, and Aydin Berenjian. 2016. “Bioconcrete: Next Generation of
Self-Healing Concrete.” Applied Microbiology and Biotechnology. Springer Verlag.
https://doi.org/10.1007/s00253-016-7316-z.

[18] Stabnikov, Viktor, Chu Jian, Volodymyr Ivanov, and Yishan Li. 2013. “Halotolerant, Alkaliphilic
Urease-Producing Bacteria from Different Climate Zones and Their Application for Biocementation
of Sand.” World Journal of Microbiology and Biotechnology 29 (8): 1453–60.
https://doi.org/10.1007/s11274-013-1309-1.

[19] Xiao, Xi, Li Xuan Goh, Cise Unluer, and En Hua Yang. 2021. “Bacteria-Induced Internal
Carbonation of Reactive Magnesia Cement.” Construction and Building Materials 267 (January).
https://doi.org/10.1016/j.conbuildmat.2020.121748.

[20] Xu, Guojing, Dongwei Li, Binquan Jiao, Dou Li, Yajie Yin, Limei Lun, Ziqiang Zhao, and Shan Li.
2017. “Biomineralization of a Calcifying Ureolytic Bacterium Microbacterium Sp. GM-1.”
Electronic Journal of Biotechnology 25 (January): 21–27. https://doi.org/10.1016/j.ejbt.2016.10.008.

573
 ERURJ 2023, 2, 4, 554-574

[21] Zaghloul, Eman H., Hassan A.H. Ibrahim, and Dalia El S. El-Badan. 2021. “Production of
Biocement with Marine Bacteria; Staphylococcus Epidermidis EDH to Enhance Clay Water
Retention Capacity.” Egyptian Journal of Aquatic Research 47 (1): 53–59.
https://doi.org/10.1016/j.ejar.2020.08.005.

[22] Bang, Sookie S., and V. Ramakrishnan. "Microbiologically-enhanced crack remediation (MECR)."
In proceedings of the international symposium on industrial application of microbial genomes, vol. 1,
pp. 3-13. 2001.
https://citeseerx.ist.psu.edu/document?repid=rep1&type=pdf&doi=f04cacaa66beabb9ecc19235ee781
bd2db158039

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