Chemical Engineering Science 238 (2021) 116610

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Chemical Engineering Science

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

Review

Bio-cement production using microbially induced calcite precipitation

(MICP) method: A review
Kbrom Alebel Gebru a,⇑, Tekleweyni Gebremicael Kidanemariam a, Haile Kidane Gebretinsae b
a
Department of Chemical Engineering, College of Engineering and Technology, Adigrat University, Tigray, Ethiopia
b
Department of Construction Technology and Management, College of Engineering and Technology, Adigrat University, Tigray, Ethiopia

h i g h l i g h t s

MICP method is a natural and biological process.

MICP is applied for strength improvement, heavy metals separation, dust control.

The effectiveness of bio-cement depends on the uniform distribution of calcite.

Further investigations to solve the basic challenges in MICP process are mandatory.

a r t i c l e i n f o a b s t r a c t

Article history: To meet the demands of highly developing construction distribution over the world, the production of
Received 11 June 2020 more amount of cement is mandatory. But, the production process of cement is highly energy consum-
Received in revised form 10 March 2021 able, environmentally unsafe, and forms crack. These issues are basic motivations to introduce new con-
Accepted 21 March 2021
struction biomaterial using the microbially induced calcium carbonate precipitation (MICP) process. It is
Available online 26 March 2021
the production of CaCO3 in the presence of Urea, CaCl2, and Sporosarcina pasteurii bacteria. The bacteria
serve as a nucleation site for Ca2+ ion attraction in its cell due to its negative charge and produces urease
Keywords:
enzyme to facilitate urea hydrolysis. Properly produced CaCO3 helps to create a strong bond between
Bio-cement
MICP
each soil particle having high unconfined compression strength and low water permeability. In this
Sporosarcina pasteurii paper, previous studies related to the production of bio-cement using MICP and factors that affect the
Energy consumable process, namely bacteria, pH, cementation solution, injection, and temperature are thoroughly reviewed.
CaCO3 Ó 2021 Elsevier Ltd. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2. Microbially induced calcium carbonate precipitation (MICP) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2.1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2.2. Literature review . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
3. Control parameters of microbially induced CaCO3 precipitation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3.1. Bacteria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3.2. Soil condition. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3.3. Cementation solution . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3.4. pH. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3.5. Temperature . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3.6. Injection system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
4. Physical and chemical properties of bio-cemented soils . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
4.1. Unconfined compressive strength (UCS) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
4.2. Calcite content. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
4.3. Water permeability . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

⇑ Corresponding author.
E-mail address: alebelkibrom@adu.edu.et (K.A. Gebru).

https://doi.org/10.1016/j.ces.2021.116610
0009-2509/Ó 2021 Elsevier Ltd. All rights reserved.
K. A. Gebru, T.iG. Kidanemariam and H.K. Gebretinsae Chemical Engineering Science 238 (2021) 116610

4.4. Urease activity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

5. Advantages and disadvantages of the MICP process . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
6. Summarized outcomes of the literature . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
7. Future research and prospects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
8. Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
Declaration of Competing Interest . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

Nomenclature

ATCC American Type Culture Collection MICP Microbially induced calcium carbonate precipitation
CFU Colony-forming unit OD Optical density
EDTA Ethylene diamine tetra-acetic acid PVC Polyvinyl chloride
EICP Enzyme-induced calcium carbonate precipitation Rpm Revolutions per minute
MAICP Microalgae-induced calcium carbonate precipitation UCS Unconfined compressive strength

1. Introduction covered by cement factories (Phua and Røyne, 2018). Therefore,

Currently, the construction industry over the world is growing
rapidly parallel to the countries’ development. To meet all the
demands of construction, more cement products are needed. The
production of cement by itself requires a huge amount of fuel as a
source of energy (Pacheco-torgal et al., 2016). Cement is the most
commonly used artificial construction material worldwide
(Chaparro-Acuña et al., 2017), which binds and helps to produce high
compressive strength between soil particles (Ivanov and Stabnikov,
2017). It increases the lifetime or durability of various infrastruc-
tures (Varalakshmi and Devi, 2014), except for its crack formation
during service time (Shivangi Mathur, 2018). But, its effect on raw
material depletion (Achal and Mukherjee, 2015), energy consump-
tion, and environmental contamination leads to search another
option to replace the soil stabilization techniques for buildings and
other infrastructures (Liu et al., 2019), where buildings are among
the most natural resource consumers (Achal and Mukherjee, 2015).
Many researchers have played a role in introducing a new
option of soil stabilization or improvement in its strength, stiffness,
permeability, and others by using construction biotechnology.
There are three basic construction biotechnology techniques called
microbially induced calcium carbonate precipitation (MICP),
enzyme-induced calcium carbonate precipitation (EICP), and
microalgae-induced calcium carbonate precipitation (MAICP). All
the techniques are natural and biological means of producing cal-
cium carbonate (CaCO3) precipitation by using urea and CaCl2 as
common ingredients and bacteria, urease enzyme, and microalgae
for MICP, EICP, and MAICP, respectively. For MICP applications,
bacteria are directly introduced as an ingredient and would grow
in ammonium-yeast media that includes ammonium sulfate, yeast
extract, and tris-buffer with the help of laboratory equipments
such as the incubator and autoclave. While in the EICP method,
urease enzyme would be produced from different plant species
such as jack beans and soybeans for bio-cement production appli-
cations. It does not use or apply any microorganisms (Kavazanjian
and Hamdan, 2015). The nontoxic urease enzyme, (Neupane et al.,
2013) has a size of 12 nm (Arab, 2019) and this smaller size makes
it preferable for all soils/sands, including fine and silt sands (Gao
et al., 2019). MAICP method is the application of microalgae as a
source of urea amidolyase enzyme to produce bio-cement
(Ariyanti and Handayani, 2011).
Almost 5% of the world’s carbon dioxide emissions from the
calcination process and fuel combustion to the environment is
2
introducing one effective method, bio-cementation (Chittoori
et al., 2018), in the cement industries is mandatory and the product
is termed as bio-cement (Phua and Røyne, 2018). It is a construc-
tion biotechnology material that acts as a binder for soil particles
to succeed in the cementation process (Phua and Røyne, 2018). It
is a green material that forms solid mass (Achal and Mukherjee,
2015) like bio-concrete (Castro-Alonso et al., 2019). It could be pro-
duced at low temperatures and preferable for in-situ systems due
to its capability to treat the injured construction within its original
position (Ariyanti and Handayani, 2011).
MICP is a construction biotechnology technique applied for
improving soil property like strength and stiffness in a natural,
easy, and sustainable way (Liu et al., 2019). It is also called bio-
calcification (Haouzi and Courcelles, 2018) and the process is per-
formed by introducing bacteria solution commonly Sporosarcina
pasteurii/Bacillus pasteurii into the prepared soil matrix with the
presence of a well-prepared chemical solution made up of urea
and calcium salts mostly calcium chloride (Kadhim and Zheng,
2016).
2. Microbially induced calcium carbonate precipitation (MICP)
2.1. Introduction
MICP is a biological method (Rahman et al., 2015), which has
been used for reinforcements or improvement properties of differ-
ent types of soils or sands (Peng and Liu, 2019). It uses ureolytic
bacteria, Sporosarcina pasteurii, that produces urease enzyme
(Arab, 2019), to hydrolyze urea to give the precipitation of CaCO3,
a thermodynamically stable material (Rahman et al., 2015). The
precipitation of CaCO3 helps to have good bonding forces within
the particle of soils (Nafisi et al., 2018). The bacteria could cultivate
under aerobic and anaerobic conditions and is active in the pres-
ence of water (Rahman et al., 2015). It consumes nutrients such
as nutrient broth, carbon, nitrogen, and others as the source of
energy to survive (Seifan and Berenjian, 2018). It has a great role
in saving the environment (free from greenhouse gas emission)
(Rahman et al., 2015), and improves soil strength in a sustainable
way (Kong et al., 2019). Its lowest cost, variety of applications
(Peng and Liu, 2019), independent of climates, and easy operations
make it a more preferable method (Chen et al., 2016).
Researchers have tried to use calcium acetate, calcium nitrate
(Abo-El-Enein et al., 2012), limestone, and eggshells as a low cost
K. A. Gebru, T.iG. Kidanemariam and H.K. Gebretinsae
source of calcium ions (Phua and Røyne, 2018). Moreover, some
studies indicated that brewery waste yeast would become a source
of nutrients for the bacteria (Chaparro-Acuña et al., 2017). All bac-
teria are photosynthetic microorganisms that could precipitate
CaCO3 in the presence of urea and calcium ions, but as they are
harmless to humans, Sporosarcina pasteurii bacteria are preferable
(Siddique and Chahal, 2011).
MICP has various applications such as sand/soil consolidating,
strength and stiffness improvement, heavy metal separation,
wastewater treatment (Achal and Mukherjee, 2015), brick manu-
facturing, slope modification or changing (Pacheco-torgal et al.,
2016), controlling or stabilizing desert sands, improving soil ero-
sion resistance (Chen et al., 2016), improving construction work
capability of weak ground soils (Ashraf et al., 2017), dust elimina-
tion or controlling (Gomez et al., 2014), greenhouse gas removal
from landfill areas (Siddique and Chahal, 2011), concrete self-
healing (Castro-Alonso et al., 2019), water permeability reduction
of concretes, and increased durability of pipes in dams (Kadhim
and Zheng, 2016).
2.2. Literature review
Many researchers have conducted studies on the precipitation
of CaCO3 using MICP through urea hydrolysis; it is easily controlled
and produces more amount of CaCO3 precipitation (Umar et al.,
2019) for a different method of soil improvement. Kong et al.
(2019) performed their study on MICP to improve the strength of
the residual soil at various temperatures (Kong et al., 2019). They
obtained the urease-producing Sporosarcina pasteurii bacteria from
American Type Culture Collection (ATCC) and it was cultivated
under the aerobic condition as described in Kong et al. (2019).
Hence, they have used a spectrophotometer to obtain concentra-
tions of 1.6  106 CFU/ml at 600 nm optical density (OD600). A
cementation reagent that was prepared by dissolving 3 g nutrient
broth, 36.8 g urea, and 15.02 g calcium chloride in 500 ml of deion-
ized water solution at 0.5 M concentrations was used as a source of
carbonate and calcium ions (Kong et al., 2019). The prepared
reagents were sprayed over the bacteria solution using the stepped
injection method for 48 h at a 6 h interval with the proportions of
2:1, 1:1, and 1:2, respectively to obtain efficient MICP treatment.
The three mixtures (soil, bacteria, and reagents) were then com-
pacted into a 38 mm diameter and 76 mm height stainless steel
mold. The specimens were then extruded and cured at atmospheric
temperatures (22 °C34 °C), 45 °C, 55 °C, and 65 °C for 1, 3, and
7 days, respectively, and finally a good shear strength of the treated
soil was obtained at temperatures of 55 °C, which is in the third
day and at a bacteria/reagents proportion of 2:1. The availability
of excess bacteria helps in rapid urease activity and in turn pro-
duces more amount of precipitated CaCO3 (Kong et al., 2019).
Furthermore, Peng and Liu (2019) conducted their experiments
to determine the influence of temperature on the strength
improvement of soils with the help of MICP through urea hydroly-
sis (Peng and Liu, 2019). Accordingly, it affects the growth and
reactivity of bacteria. The experiments were conducted in an aque-
ous solution and sand column at different temperature intervals
(10 °C, 15 °C, 20 °C, 25 °C, and 30 °C). The precipitated CaCO3
reduces the porosity and permeability of the soil. The Sporosarcina
pasteurii bacteria were grown in a culture media at 30 °C for 40 h
aerobically, and then the final cultivated bacteria were stored at a
temperature of 4 °C. The cementation media (urea-CaCl2) was pre-
pared with a ratio of 1.0 M and 0.5 M, respectively, and Ottawa sil-
ica sand (round shape) was used with a density of 1.63 g/cm3 (void
ratio of 0.63). A mixture of bacteria solution (80 ml) and cementa-
tion media (1440 ml) were mixed with 2 L aqueous solution and a
magnetic stirrer was used for uniform mixing. The formed Ca2+ was
measured by the EDTA (ethylene diamine tetra acetic acid)
3
Chemical Engineering Science 238 (2021) 116610
titration method to calculate the precipitated amount of CaCO3
(Peng and Liu, 2019).
As shown in Fig. 1 below, the column was made up of polyvinyl
chloride (PVC) tubing with a diameter of 47 and a length of
100 mm and covered with 10 mm gravel at both sides to have a
clean reaction and a peristaltic pump was used to pump the solu-
tion throughout the specimen. Initially, the sand was mixed with
0.05 mol/l CaCl2 (75 ml volume) before 75 ml bacteria solution
was poured and was followed by cementation media after a 12-h
interval and this process was repeated for 6–20 cycles. The ure-
olytic activity increases with increasing temperatures, but it starts
to decline as the temperature highly increases while the amount of
the precipitated CaCO3 increases with decreasing temperature as
shown in Table 1. Finally, the authors concluded that the amount
of CaCO3 precipitated in the aqueous solution was better than sand
columns at 10 °C than 30 °C (Peng and Liu, 2019).
The effect of grain size and shape on the mechanical behavior of
MICP sand was investigated by Nafisi et al. (2018). Three shapes of
sand, such as angular coarse, round coarse, and round fine, were
taken as samples, and Sporosarcina pasteurii bacteria were culti-
vated as described by Nafisi et al. (2018). After the optical density
(OD600) was recorded at 0.8–1.2, it was centrifuged at 4000 g for
around 15 min. The final cultivated bacteria were stored at 4 °C
for around 14 days (Nafisi et al., 2018) (Fig. 2). Using the two-
phase injection method (minimizes the formation of flocculation
and clogging of the pore voids near the injection end, (Cheng
et al., 2018), the biological solution (urea 333 mM and ammonium
chloride (NH4Cl) 374 mM) at a rate of 10 ml/min followed by the
cementation solution (urea 333 mM, NH4Cl 374 mM, and calcium
chloride of 100 mM) was introduced to the specimen. Before the
cementation solution was injected, the specimen contained sand
and Sporosarcina pasteurii bacteria, and the biological solution
was kept for 4–6 h to have enough time for the bacteria to attach
to the sand particles. Then, with the 6-6-12 time intervals, the two-
pore volume of cementation was injected. The result indicated that
the efficiency of the CaCO3 was increased with the fine particles
even at a small amount of CaCO3 precipitation (Nafisi et al., 2018).
Cheng et al. (2018) investigated the soil bio-cementation using
a new one-phase low-pH injection method. The all-in-one solution
Fig. 1. Setup for the sand column treatment (Peng and Liu, 2019).
K. A. Gebru, T.iG. Kidanemariam and H.K. Gebretinsae Chemical Engineering Science 238 (2021) 116610

Table 1
Effect of temperature in precipitated calcite content (Peng and Liu, 2019).

Run Temperature (°C) Corresponding Calcite content (g)

1 10 40.6
2 15 32.5
3 20 25.2
4 25 23.1
5 30 21.1

was prepared by mixing the cultivated Sporosarcina pasteurii bacte-

ria, deionized water, and cementation solution (2 M urea and
CaCl2), which is 1 M of the total at the initial of 4 pH value. The
PVC sand columns that have a diameter of 50 mm and length of
120 mm were packed with silica sand with various particle sizes
and then, the solution (90 ml) was injected once using the peri-
staltic pump from bottom to top with a rate of 1 L/h until the spec-
imen saturated at room temperature of 25 °C. The injection was
repeated four times to have good cementation for every 24 h. Sam-
ples with a ratio of 1:1.5 and 1:2 diameter in height were taken to
measure the strength and uniformity of the precipitated CaCO3.
The results indicated that the conversion efficiency decreases with
a decrease in urease activity. By increasing the number of injec-
tions and giving enough time, it increases the cementation level,
which in turn, strengthens the column (Cheng et al., 2018).
Furthermore, Animesh Sharma (2016) studied the effect of
microbial-induced calcite precipitates on the strength of fine-
grained soils (Animesh Sharma, 2016). Sporosarcina pasteurii bacte-
ria with concentrations of 1  105, 1  106 and 1  107 CFU/ml and
cementation reagent (equal amount of urea, CaCl2, and 3 g/l of
broth) with 0.25 M, 0.5 M, 0.75 M, and 1.0 M concentrations were
prepared. The Sporosarcina pasteurii bacteria solution was first
mixed with the soil before the cementation solution was injected.
Samples were kept at a temperature of 20 °C 30 °C for 0, 3, and
7 days. Higher unconfined compression strength was found at high
treatment duration time, 0.5 M, and 1  106 CFU/ml concentration
of reagents and bacteria, respectively. It was possible to produce
more bacteria at low cost (Animesh Sharma, 2016). Nemati et al.
(2005) were the leaders of permeability profile modification who
used bacterially formed CaCO3 in comparison to enzymatic option
(Nemati et al., 2005). The cementation solution with a ratio of 1:2.5
urea and CaCl22H2O was prepared. As seen from Fig. 3, three glass
columns with an inner diameter and height of 2.5 and 30 cm,
respectively, were prepared. The glasses were filled by sand and
glass beads with an average diameter of 225 and 75 mm until
70/30 percentage, respectively. A glass tube was used to maintain
the head pressure. The desired Sporosarcina pasteurii bacteria were
cultivated in a medium as described in Nemati et al. (2005). More-
over, the mixed sample was maintained at 10, 20, and 30 °C to
Fig. 3. Experimental setup to strengthen the sand (Nemati et al., 2005).
show the effect of temperature. At a high amount of urea and cal-
cium chloride, the concentration of CaCO3 produced was almost
three times lower in the presence of bacteria than the enzymatic
condition, and this was due to the effect on the bacteria reproduc-
tion. In the range of 20 °C–30 °C, more CaCO3 precipitation was
obtained. But, above this temperature, it was highly decreased in
the enzyme option, because the enzyme has low resistance capac-
ity to temperature rising than bacteria (Nemati et al., 2005).
In a study conducted by Umar et al. (2019), the maximum
unconfined compressive strength (UCS) was obtained at 48 h. But
beyond this time, it was reduced due to urease activity reduction.
Moreover, calcite content also declined beyond 48 h due to the
lowering of bacterial activity (Umar et al., 2019). During the bio-
chemical reactions in the presence of urease activity, hydrolysis
of 1 mol of urea gives a corresponding amount of ammonia and
carbamate (NH2COOH). Carbamate, in turn, hydrolyses to ammo-
nia and carbonic acid (Achal and Mukherjee, 2015; Piriyakul and
Iamchaturapatr, 2013). The effectiveness of bio-cement depends
on the distribution of CaCO3 deposition across the soil matrix
and the amount of ammonia released to the environment could
be minimized by applying an optimized amount of urea (Ashraf
et al., 2017; Piriyakul and Iamchaturapatr, 2013). The chemical
processes through urea ureolysis are presented in Fig. 4 below.

Fig. 2. Treated specimens with different sand sizes (Nafisi et al., 2018).

4
K. A. Gebru, T.iG. Kidanemariam and H.K. Gebretinsae Chemical Engineering Science 238 (2021) 116610

Fig. 4. Chemical processes through urea ureolysis (Piriyakul and Iamchaturapatr, 2013).


Moreover, the overall reaction occurs inside the particular sys- CaCl2 ! Ca2þ þ 2Cl ð6Þ
tem, forms bio-cement in the presence of the alkalophilic bacteria,
which is presented below (Achal and Mukherjee, 2015). This bio- Ca2þ þ Cell ! Cell  Ca2þ ð7Þ
cement form of calcite was a more stable form of CaCO3, while
aragonite was metastable and was changed to calcite after a time
Cell  Caþ2 þ CO2
3 ! Cell  CaCO3 ð8Þ
(Al-Thawadi, 2011).
In reaction (1), urea was decomposed or hydrolyzed to carba- 
mate and ammonia. The pH value of the whole system would be 2NHþ4 þ 2Cl ! 2NH4 Cl ð9Þ
raised due to the formation of ammonium ion in reaction (4) and Furthermore, Sun et al. (2018) conducted their investigation on
as shown in reaction (7) the bacteria cell was served as a nucle- the improvement of MICP technology for sand solidification (Sun
ation site for Ca2+ and finally, NH4Cl was released as an effluent et al., 2018). The Sporosarcina pasteurii bacteria was grown in a
(Al-Thawadi, 2011). medium as described in Sun et al. (2018). As shown in Fig. 5, ster-
COðNH2 Þ2 þ H2 0 ! NH2 COOH þ NH3 ð1Þ ilized sand with a diameter of less than 0.5 mm was packed with
the bacteria solution (flow rate of 1 ml/min with the help of an
NH2 COOH þ H2 0 ! NH3 þ H2 CO3 ð2Þ electric pump) in the PVC cylinder, which has an inner diameter
and height of 4.6 and 20 cm, respectively, then left until two hours.
H2 CO3 ! HCO3 þ Hþ ð3Þ They prepared three samples with different quantities of sand such
as 210, 230, and 290 g and with their corresponding 1, 2, and 4 ml/
2NH3 þ 2H2 O ! 2NHþ4 þ 2OH ð4Þ min injection speed. High urease activity was obtained when urea
was added after sterilization and high strength was also found
within high injection speed regardless of the sand column height.
HCO3 þ Hþ þ 2NHþ4 þ 2OH ! CO2 þ
3 þ 2NH4 þ 2H2 O ð5Þ
The bacterium was more active in the alkaline environment with

Fig. 5. Sand solidification tests in PVC cylinders with 210, 230, and 290 g of sand and with 1, 2, and 4 ml/min injection speed (Sun et al., 2018).

5
K. A. Gebru, T.iG. Kidanemariam and H.K. Gebretinsae
a pH of 9–10; and good urease activity was found at 5–20 g/l urea
amount; more amount of urea leads to more alkalinity condition
and this was beyond the resistance ability of the bacteria (Sun
et al., 2018).
As Cheng et al. (2014) reported in their study on the bio-
cementation of sandy soil using MICP for marine environments,
seawater could be a good resource for calcium ions (Cheng et al.,
2014). They have prepared two samples to compare the soil stabi-
lization by the MICP method using seawater and calcium chloride
as a source of calcium ion. In both samples, pure silica sand (coef-
ficient permeability: 1.74  104 m/s) with a variety of diameters
was packed to the PVC column with 50 and 180 mm inner diame-
ter and height, respectively. Seawater cementation (seawater and
0.6 g/l or 10 mM urea) and cementation solution (1 M, 60 g/l urea
and 1 M, 111 g/l anhydrous CaCl2) were added to the soil next to
the bacterium culture, Sporosarcina pasteurii, (85 ml) at 25 ± 1 °C.
For both solutions, the bio-cementation reaction of 6 and 24 h were
given, respectively. The compressive strength of samples treated
with seawater was higher than that of samples untreated with
the cementation solution. A high permeability improvement,
which was 40 times higher, was found as compared to untreated
samples. In general, the MICP process using seawater has a low
rate of calcite precipitation but reduces the usage of additional
chemicals as a source of calcium ions (Cheng et al., 2014). In addi-
tion, Kim et al. (2018) studied the effect of temperature, pH, and
reaction duration on MICP in comparison to two microbial species
Staphylococcus saprophyticus and Sporosarcina pasteurii. Different
samples were prepared to test the effect of pH, temperature, and
time. The authors have conducted the experiments in pH (6, 7, 8,
9, and 10), temperature (20, 30, 40, and 50 °C), and the duration
time of 2, 3, and 4 days. Fig. 6 given below shows the preparation
and amount determination of CaCO3 in solution form.
The pressure exerted by CO2 gas would be measured to deter-
mine calcite precipitation. The results indicated that the amount
of the calcite precipitate was higher using S. saprophyticus than S.
pasteurii bacteria. But the optimum conditions for both bacteria
were found to be at a pH of 7, the temperature of 30 °C, and dura-
tion time of 3 days (Kim et al., 2018).
Fig. 6. Experimental process for measuring the calcite precipitation (Kim et al., 2018).
6
Chemical Engineering Science 238 (2021) 116610
Cheng et al. (2013) also studied on the cementation of sand soil
MICP at various degrees of saturation and have found better soil
strength and stiffness at a low degree of saturation. Pure silica sand
with various diameters and Bacillus sphaericus bacteria, cultivated
as described by Cheng et al. (2013), and urease activity of 10 U/
ml were used for experimentation. The sand column of the PVC
consisted of 160 and 55 mm length and inner diameter, respec-
tively, and the vacuum pump was used to remove the excess solu-
tion. A bacterial solution was injected with a flow rate of 1 L/h and
followed by a cementation solution after 12 h at 25 ± 1 °C. Final
samples were taken and dried for 24 h at 60 °C to investigate the
results, but it would be stored for 7 days at 20 ± 1 °C before char-
acterization. Irrespective of saturation degrees, the permeability
decreases for both fine and coarse sand particles. The permeability
of bio-cemented samples was highly reduced than in samples trea-
ted with Portland cement (Cheng et al., 2013).
Zhao et al. (2014) used bacteria and enzymes to evaluate the
factors affecting the treated soil specimens (Zhao et al., 2014).
The medium particle size of 0.46 mm Ottawa silica sand was used.
The Sporosarcina pasteurii bacteria were grown in the media as
described in Zhao et al. (2014) until 107-108 cell/ml concentrations.
The treated bacterium media were then kept at 4 °C until used.
After 85 ml of bacteria or the enzyme solution was mixed with
330 g sand, the cementation media (urea and CaCl2 with different
concentrations) were poured. The results indicated that as the
OD600 increased from 0.3 to 0.6, the UCS was increased by 200%
from 0.44 to 1.36 MPa. In the case of urease concentration increas-
ing from 5 to 15 g/l, it provides a high amount of CaCO3 precipita-
tion when it was cured at room temperature and the treated sand
becomes stable and with no significant effect on the environment.
In the same amount of CaCO3, UCS in samples treated with bacteria
were always greater than those samples treated with urease
enzymes and this was due to the lower efficiency of the enzyme,
which resulted in the degradation of the urease enzyme (Zhao
et al., 2014).
According to Al Qabany et al. (2012), the grain size of sand
between 50 and 400 mm was preferable for the MICP process (Al
Qabany et al., 2012). Bacteria could not freely move across the soil
K. A. Gebru, T.iG. Kidanemariam and H.K. Gebretinsae
matrix below this size. To date, it was mostly performed at the lab-
oratory scale, but some efforts were made to scale up to the field
scale (Soon et al., 2014). This method was tried for a larger scale
by using 0.5 M urea-CaCl2 concentration with a rate of 10 ml∕min
for 12 h with the help of a confining pressure of 20 KPa. When the
urease activity was very high, CaCO3 would be precipitated in the
form of vaterite, which needed time to change to the calcite form.
The rate of urea hydrolysis by bacteria was not directly implied due
to the rate of calcite precipitation because there were many factors
affecting the process (Al Qabany et al., 2012). A large amount of
precipitated calcite was required for coarse soils to have good
strength improvement (Soon et al., 2014). Moreover, during proper
and efficient CaCO3 precipitation systems, the amount and crystal
size of CaCO3 precipitate in seawater was greater than soft water.
This is due to the presence of more calcium ions in seawater
(Varalakshmi and Devi, 2014).
As many researchers indicated, the optimum temperature con-
ditions for urea hydrolysis in the presence of urease enzyme or
other microorganisms were found between 20 °C and 37 °C. It
was also reported that the amount of CaCO3 precipitation could
not increase with the increase of urea and Ca2+ concentration
above 36 and 90 g/l, respectively (Okwadha and Li, 2010). Thus,
according to Okwadha and Li (2010)’s report, the optimum condi-
tions for CaCO3 precipitation using MICP were found to be 250 mM
Ca2+, 666 mM urea, and bacterial cell concentration of 2.3  108
cells/ml (Okwadha and Li, 2010). The precipitated CaCO3 fills the
pore gap or space rather than forming a bridge and this phe-
nomenon reduces the hydraulic conductivity or permeability of
the treated samples (Nayanthara et al., 2019). Besides, the point-
to-point bond creation between CaCO3 precipitations increases
the strength behavior of the treated specimens (Chu et al., 2014).
Strength improvement of coarse materials was not relatively
cost-effective due to their acquirement of more cementation solu-
tions to precipitate more calcite crystals (Mahawish et al., 2017)
and the spherical/ellipsoidal particle morphology of the precipi-
tated calcite leads to having more contact between soil particles,
which in turn, forms high bond strength (Qian et al., 2019).
3. Control parameters of microbially induced CaCO3
precipitation
For the efficient and effective method of producing CaCO3 pre-
cipitation, which is a basic cementing material (Terzis and Laloui,
2019), various parameters should be controlled during the process,
such as bacteria, soil, cementation solution, pH, temperature, and
injection system.
3.1. Bacteria
Bacteria, in general, are used to facilitate the hydrolysis of urea
to carbon dioxide and ammonia and act as a nucleation site for Ca2+
collection in its cell due to its high negative charge. This negative
charge is due to rising pH levels within the solution. Among the
various kinds of bacteria, Sporosarcina pasteurii/Bacillus pasteurii
is preferable because of its high urease activity, ability to grow at
pH above 8.5 (Ivanov and Stabnikov, 2017), and its relatively good
resistance to the ammonia effect (Chaparro-Acuña et al., 2017). Its
cell served as a nucleation site for CaCO3 formation (Cui et al.,
2017). The concentration of the Sporosarcina pasteurii bacteria used
is directly proportional to the CaCO3 precipitated, the higher the
bacterium higher will be the CaCO3 precipitate. The size of all the
bacteria ranges between 0.5 and 3.0 mm. But, the size of the mostly
prefered bacteria bacteria, Sporosarcina pasteurii, is 1 mm (Kadhim
and Zheng, 2016). These bacteria do not affect the environment
(Haouzi and Courcelles, 2018). A bacterium concentration between
7
Chemical Engineering Science 238 (2021) 116610
1  106 and 1  108 cells is required to precipitate a high amount of
CaCO3 (Chaparro-Acuña et al., 2017).
3.2. Soil condition
The size of the soil pores should be allowed for bacteria to freely
move inside it. It has a great effect on MICP and is preferable when
the size is between 50 and 400 mm (Kadhim and Zheng, 2016). For
any sands/soils washing using deionized water before treatment
makes it free from salts and other impurities (Liu et al., 2019). Dif-
ferent types of soil precipitate a different amount of CaCO3 precip-
itation or deposition. This difference is due to the sand or soil
behavior in its temperature, size (sands/soils with small size parti-
cles are more preferable and for the coarse particles more amount
of precipitated CaCO3 is required to have the desired strength), and
shape of particles (spherical/ellipsoidal particle morphology is nec-
essary) (Inagaki et al., 2011).
3.3. Cementation solution
The chemical solutions prepared have a great effect on the
amount of CaCO3 precipitation. Having a higher amount of Urea-
CaCl2 solution helps for an efficient MICP process. The effective
preparation of cementation solution lead to having more calcite
precipitation in every gap or space and the surrounding of the par-
ticles in turn creates overall strength. Besides, the presence of
enough amount of chemical solution within the particular system
decreases the water permeability of the treated specimens (Al
Qabany and Soga, 2013). Some researchers indicated that the con-
centration of cementation media between 0.25 and 0.5 M gives
good precipitation but beyond this amount, it would not be fully
utilized and has less effect on the precipitation amount of CaCO3
and strength (Zhao et al., 2014).
3.4. pH
CaCO3 precipitation using the MICP process is more effective in
alkaline or a weak basic environment. Some bacteria could produce
CaCO3 in acidic conditions or pH of less than 7, but because they
are harmful to humans, they are not preferable. In many studies,
the optimum level of pH for the urease enzyme was found to be
8 (Soon et al., 2014). For the Sporosarcina pasteurii bacteria, it could
be raised to 8.7–9.5 (Kim et al., 2018). The increasing pH value in
the solution is due to the formation of ammonium ion (NH+4) from
urea hydrolysis in the presence of Sporosarcina pasteurii bacteria
(Kim et al., 2018). Urease activity decreases as the pH value
increased above 10 (Torres-Aravena et al., 2018). Having optimized
pH in the process facilitates the formation of CaCO3 and absorbs
CO2 gas to the solution due to the presence of OH (Li et al., 2013).
3.5. Temperature
Temperature is one basic factor that could affect the whole pro-
cess and efficiency of the formed CaCO3 in any application. The
produced CaCO3 in the same amount of chemical solution, duration
time, and pH but in different temperatures could not have the
same strength; this is due to the difference in crystal size of the
precipitated calcium carbonate. Thus, having a higher amount of
precipitated CaCO3 in a system does not mean that it has good
strength (Kim et al., 2018). As many investigators reported, the
optimum temperature range for the efficient and maximum pre-
cipitation of the CaCO3 varied between 20 °C and 37 °C and there
was no reaction when the temperature was below 5 °C (Kadhim
and Zheng, 2016). At room temperature, the formed calcium car-
bonate, which is a calcite crystal exists in a stable form (Shivangi
Mathur, 2018). According to Chaparro-Acuña et al. (2017), urease
K. A. Gebru, T.iG. Kidanemariam and H.K. Gebretinsae
activity becomes stable at 35 °C but decreases by almost 47% when
it increased to around 55 °C.
3.6. Injection system
There are various ways of injection of the cementation solution
into the specified soil type. The way of injection solution has its
effect on the amount and efficiency of calcite precipitation. The
non-uniform distribution of CaCO3 within the column length is
due to improper mixing or injecting of bacteria solution, bacteria
collection in a specific area (Ng et al., 2012). Mainly there are four
ways of injecting the solutions:
a. Two phases: two phases are the way of injecting the Sporo-
sarcina pasteurii bacteria solution to the specimen before the
cementation solution was injected and helps to minimize
the formation of flocculation and clogging of the pores
(Cheng et al., 2018). It is a more preferable way of the injec-
tion system (Shivangi Mathur, 2018).
b. Three-phase injection: initially, the Sporosarcina pasteurii
bacteria is injected to the soil matrix before CaCl2 injection
and followed by cementation solution (urea, source of cal-
cium, and other nutrients if any) (Arab, 2019).
c. Staged injection or stopped/stepped: this method gives
time for the Sporosarcina pasteurii bacteria solution before
injecting the cementation solution to have a uniform distri-
bution of precipitated CaCO3 (Cheng et al., 2018; Ng and
Min-Lee Lee, 2012).
d. Multiple phases/single-stage injection: is injecting the
solution at once and it is difficult to predict the response
in addition to complexity (Cheng et al., 2018). Since all the
solutions are injected simultaneously, there would be clog-
ging at the entry point. Thus, it is considered as less effective
method (Mahawish et al., 2018).
4. Physical and chemical properties of bio-cemented soils
4.1. Unconfined compressive strength (UCS)
UCS is used to measure the strength of the treated specimen
and is related to the amount of CaCO3 precipitated when every pro-
cess parameters such as temperature, pH, injection systems, and
concentrations are well controlled and applied efficiently (Haouzi
and Courcelles, 2018). UCS is highly dependable on the uniform
distribution of the precipitated CaCO3 throughout the selected
specimen exposed to the test. Improper or non-uniform distribu-
tion of calcite form of CaCO3 in the sample forms cracks and tends
to easily break. This random distribution is maybe due to the inef-
ficient use of the above process parameters. UCS tests are con-
ducted on the bio-cemented soil specimens after it is dried at
50 °C for 24 h with selected diameter-to-height ratios and with a
constant rate of axial load (Cheng et al., 2014).
4.2. Calcite content
Calcite content is determined by using the gravimetric analysis
of acidified samples in two ways. First, 10 g of the powder sample
can be used after oven drying at 105 °C for 24 h. Then 2 M
hydrochloric acid can be added to the prepared powdered sample
and carbon dioxide would liberate due to the reaction between
calcite and hydrochloric acid. The residue would be collected and
oven-dried again, and the loss in weight before and after acid
washes (Mahawish et al., 2018), would be used to estimate the per-
centage of calcite contents in the specimen (Umar et al., 2019). Sec-
ond, since the applied pressure by carbon dioxide read on the
gauge is proportional to the calcite form of CaCO3 formed; it needs
8
Chemical Engineering Science 238 (2021) 116610
the direct application of hydrochloric acid to powder CaCO3 and
this generates CO2 gas, which in turn creates a pressure difference
on the system. This pressure difference shows the amount of cal-
cite formed. The process parameters that give the maximum calcite
content is the optimized process (Kim et al., 2018).
4.3. Water permeability
MICP reduces the water permeability of the concrete by clog-
ging of the porous, micro-cracked materials due to the presence
of insoluble calcite in the surface (Achal and Mukherjee, 2015).
The water permeability of samples highly relates to its strength.
Having low water permeability leads to having high strength.
The permeability of treated specimens is decreased when there is
more and a uniformly distributed calcite form of calcium chloride.
Thus, all soil/sand particles are attracted or closed together to fill
the space between them. It would be measured using the constant
head permeability test at room temperature after being flushed
with deionized water to remove impurities (Mahawish et al.,
2018).
4.4. Urease activity
Urease activity is tested by measuring the difference in electri-
cal conductivity per minute of the solution, and it is proportional to
the amount of urea hydrolyzed. The bacterial solution is mixed
with the urea solution and the electrical conductivity can be mea-
sured within the time interval at a specified temperature (Sun
et al., 2018). Ureolytic activity affects the temperature difference
within the system and this results in a different measurement of
ureolytic activity. Indeed, the ureolytic activity of Sporosarcina pas-
teurii bacteria is increased with increasing temperature, but mostly
this is until the temperature of less than 50 °C. It is also affected by
time; having the same temperature but different curing time gives
different ureolytic measurements. As curing time increases, the
ureolytic activity of bacteria decreases (Peng and Liu, 2019).
5. Advantages and disadvantages of the MICP process
The MICP process is a natural (Wang et al., 2017a) and new
innovation of soil property enhancement such as strength and
water permeability. Thus, it has many advantages such as it is a
sustainable method of bio-cement production; it is free from
greenhouse gas emission; it is low-cost when compared with
cement production process; the CO2 generated from urea hydroly-
sis converts to bicarbonate ion (HCO3); it requires low temperature
for bio-cement production and eliminates coal requirement as a
fuel (Pacheco-Torgal et al., 2015; Nayanthara et al., 2019); easily
applicable for in-situ applications through CaCO3 precipitation
(Stabnikov et al., 2015; Nayanthara et al., 2019); its applicability
could not be affected by seasonal variation like winter and summer
(Shivangi Mathur, 2018); and it has versatile industrial applicabil-
ity such as mortar and bio-concrete preparation and dust control
that are harmful to the overall health of humans (Mahawish
et al., 2017; Shivangi Mathur, 2018).
Along with the above mentioned main advantages of MICP pro-
cess, it has also some limitations or disadvantages such as (1) the
pollution of the aquatic environment that harms humans and ani-
mals health due to the presence of high concentrations of ammonia
in the form of NH4Cl as effluent or by-product (Pacheco-Torgal
et al., 2015; Wang et al., 2017b) and the corrosion of construction
steel may occur. The production of bio-concrete in large scale using
MICP process could expect to contaminate 100 km3 of air and
4.5  106 m3 of drinking water. But, it could be minimized through
effluent treatment before being disposed to the environment
K. A. Gebru, T.iG. Kidanemariam and H.K. Gebretinsae
Table 2
Summarized outcomes of the literature.
Inputs
6

Sporosarcina pasteurii bacteria at the concentration of 1.6  10 CFU/ml;
cementation reagent (3 g nutrient broth, 36.8 g urea, and 15.02 g cal-
cium chloride) of 0.5 M; and samples were cured at (22 °C34 °C),
45 °C, 55 °C temperatures, and 65 °C for 1, 3, and 7 days

Sporosarcina pasteurii bacteria 80 ml and Ottawa silica; cementation
media of 1440 ml was prepared and mixed with 2 L aqueous solution;
different temperature intervals 10 °C, 15 °C, 20 °C, 25 °C, and 30 °C have
been evaluated

Sporosarcina pasteurii bacteria and silica sand; cementation solution of
90 ml was taken and repeated four times; and room temperature of
25 °C and 4 pH

Sporosarcina pasteurii bacteria with concentrations of 1  105, 1  106,
and 1  107 CFU/ml; cementation reagent with 0.25 M, 0.5 M, 0.75 M,
and 1.0 M concentrations; and samples were kept at a temperature of
20 °C–30 °C for 0, 3, and 7 days

Sporosarcina pasteurii bacteria and sand with average diameter of
225 mm; samples were maintained at 10 °C, 20 °C, and 30 °C of temper-
ature; and reagents with different concentrations of urea and CaCl2 (5,
12.5; 10, 25; 15, 37.5 g/L) were prepared

Staphylococcus saprophyticus and Sporosarcina pasteurii bacteria with
2 ml volume; samples were conducted at pH of 6, 7, 8, 9, and 10, tem-
perature of 20 °C, 30 °C, 40 °C, and 50 °C, and the duration time of 2, 3,
and 4 days
Main factors affecting the MICP process

Bacteria: Sporosarcina pasteurii/Bacillus pasteurii is preferable because of its high urease activity, ability to grow at pH above 8.5, and its
relatively good resistance to the ammonia effect

Cementation solution: concentration of cementation media between 0.25 and 0.5 M gives a good amount of CaCO3 precipitation and
high strength

pH: For the Sporosarcina pasteurii bacteria, urease activity becomes high and relatively constant in between 8 and 10 pH

Temperature: the optimum temperature range for the efficient and maximum precipitation of the CaCO3 was laid between 20 °C and
37 °C
(Castro-Alonso et al., 2019). (2) brittleness of CaCO3, calcite, and
crystal bonds formed between soil particles (Pacheco-Torgal
et al., 2015).
6. Summarized outcomes of the literature
This paper reviewed selected articles on the production of bio-
cement by using MICP through urea hydrolysis for different appli-
cations. Thus, these articles that are included and reviewed above
in this paper have been summarized in tabular form as shown in
Table 2 below.
7. Future research and prospects
The selected Sporosarcina pasteurii bacterium is only applicable
for soils/sands that have a size less than one micrometer. This
problem can be solved by introducing other types of bacteria with
smaller size by making it dead (harmless to humans) but active in
its ureolytic activity. From the literature, the cultivation of bacteria
for industrial-scale production of bio-cement is difficult to handle.
Consequently, this situation may force us to further investigate or
to replace bacteria by the direct use of urease enzyme produced
from different plants in the laboratory. Almost all articles relating
to this area could not specify or give any information about the
possibilities of reaction happening in the effluent tank, because
there is a possibility of transferring calcium ion, urea, and bacteria
as effluents. This condition may expose to additional unnecessary
by-products in an effluent tank. The overall investigation on MICP
could not indicate whether the urea, calcium ion, and bacteria are
recycled for further process as a feed or not. Besides, it could not
describe the shelf life of the bacteria after the process has been
accomplished. Further investigation is also required to briefly
Chemical Engineering Science 238 (2021) 116610
Outcomes References

High UCS at temperatures of 55 °C and in the third day (Kong et al., 2019)

Lowest calcite content and bacterial or urease activity at
atmospheric temperature

UCS of samples was decreased with increasing tempera- (Peng and Liu, 2019)
tures from 10 °C–30 °C

High urease activity but decreased calcite content was
obtained at 20 °C–30 °C than 10 °C–15 °C

Calcite content and high UCS were obtained as injection (Cheng et al., 2018)
cycle was increased

High calcite content and high UCS were found in 3 days, (Animesh Sharma, 2016)
0.5 M, and 1  106 CFU/ml concentration of reagents and
bacteria, respectively.

At temperatures of 20 °C30 °C more CaCO3 was (Nemati et al., 2005)
precipitated

Calcite amount precipitated was highly decreased in
enzymatic option at a temperature above 30 °C than
the bacteria option

High calcite content was obtained/precipitated by using (Kim et al., 2018)
Staphylococcus saprophyticus than Sporosarcina pasteurii

Accordingly, for both bacteria pH7, temperature 30 °C,
and 3 days duration time become the optimized
parameters
(Ivanov and Stabnikov,
2017; Chaparro-Acuña
et al., 2017)
(Zhao et al., 2014)
(Kim et al., 2018)
(Kadhim and Zheng,
2016)
describe the interactions between the soil, microorganism, or bac-
teria and the precipitated calcium carbonate. As many researchers
indicated that there are other alternative sources of calcium ions
such as eggshell, seawater, and limestone. Thus, it is possible to
reduce the source of chemicals and corrosions within the specific
application and minimize the cost of the overall process. The low
viscosity and brittleness behavior of the produced MICP solution
and calcite crystal needs to be improved using admixtures and/or
composite materials. Appropriate cost determination of MICP pro-
cess (cost of urea, selective bacteria, and calcium chloride) needs to
be determined to indicate the feasibility of the process comparing
with the application of cement and others. Ammonium ion or
ammonia, by-product of MICP process, is one of the aquatic ecosys-
tem polluting chemicals. But, this issue could be resolved by con-
verting ammonia to fertilizer for agricultural activities. Until
now, the bio-cement production through MICP mechanism for dif-
ferent applications was exercised in the micro level or laboratory
scale. However, there were few implementations in the field scale
by some researchers. Nevertheless, for the current development of
constructions, it should be scaled up to the mega level or industrial
scale by overcoming some challenges such as the concentration of
cementation solutions (urea and calcium chloride) and difficulty of
bacteria cultivation at the industrial level. Therefore, it needs fur-
ther and scientific investigations to solve the above basic chal-
lenges in the MICP process.
8. Conclusion
The MICP method is a natural and biological process of produc-
ing alternative construction bio-material, bio-cement, at very low
temperature, and low cost and in an environmentally friendly
manner (free of greenhouse gas emissions). The effectiveness of
9
K. A. Gebru, T.iG. Kidanemariam and H.K. Gebretinsae
the bio-cement is highly dependent on the uniform distribution of
the produced CaCO3 on the desired sample. The Sporosarcina pas-
teurii bacterium used in MICP application is active in water, harm-
less to human’s health, and has high resistance to the effect of
generated ammonia or ammonium ion. One basic drawback of
using this bacterium is, its inapplicability for sands/soils that have
less than 1 mm size, and this is due to the size of Sporosarcina pas-
teurii bacteria. MICP has various applications such as soil strength
improvement, heavy metal separation, dust control, and the reduc-
tion of soil erosion. In this paper, previous research studies related
to the production of bio-cement using MICP and all factors affect-
ing the process, namely bacteria, pH, cementation solution, injec-
tion, and temperature are thoroughly reviewed. The observations
from the literature indicated that the effectiveness of bio-cement
depends on the uniform distribution of calcite, and this happens
due to proper preparation and mixing of the cementation solution
with sands. Because of the effluent or by-product of MICP, NH4Cl,
aquatic environments may become polluted. However, this would
be minimized when ammonia was reprocessed as an agricultural
fertilizer. So far, bio-cement production through MICP method
for different applications was attempted at the micro level or lab-
oratory scale. However, there were few implementations in the
field scale by some researchers. Nevertheless, for the current devel-
opment of constructions, it should be scaled up to the mega level
or industrial scale by overcoming some challenges such as the con-
centration of cementation solutions (urea and calcium chloride)
and difficulty of bacteria cultivation at the industrial level. There-
fore, further scientific investigations are required to solve the
above basic challenges in the MICP process.
Declaration of Competing Interest
The authors declare that they have no known competing finan-
cial interests or personal relationships that could have appeared
to influence the work reported in this paper.
Acknowledgments
Messebo Cement Factory PLC. (MCF), Tigray, Ethiopia is grate-
fully acknowledged by the authors for funding their research work.
References
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