International Journal of Geosynthetics and Ground Engineering (2021) 7:78

https://doi.org/10.1007/s40891-021-00323-5

ORIGINAL PAPER

Improvement of Strength Behaviour of Residual Soil via Enzymatically

Induced Calcite Precipitation
Muttaqa Uba Zango1,2 · Khairul Anuar Kassim1 · Kamarudin Ahmad1 · Abubakar Sadiq Muhammed1,3

Received: 5 November 2020 / Accepted: 9 October 2021 / Published online: 23 October 2021
© The Author(s), under exclusive licence to Springer Nature Switzerland AG 2021

Abstract
Enzymatic induced calcium carbonate precipitation (EICP) is a biomediated soil improvement technique that utilizes free
enzyme to produce a biocement material. This research investigated by performing test tube test the effect of varying urea-
CaCl2 concentrations from 0.25 to 1.25 M on the mass and efficiency of calcium carbonate precipitation formed via EICP
technique. The study also evaluated, by conducting UCS test, the strengths of EICP-treated residual soil prepared at various
concentrations of cementation solution and different moulding water contents ( – 2% to + 4% OMC). The test tube test results
showed that the mass of calcium carbonate precipitation increased with an increment in the urea-CaCl 2 concentrations from
0.25 to 1.00 M. Maximum mass of calcium carbonate precipitation of 1.577 g, 1.603 g and 1.603 g were obtained due to
1.00 M urea-CaCl2 at 3, 7 and 14 days, respectively. The effect of curing period beyond 3 days were found to be insignificant
on the amount of ­CaCO3 precipitations. The strength of the EICP-treated soil increased with increasing concentrations of
urea-CaCl2 and decreased with an increment in moulding water content. The maximum strengths values at all moulding
water contents were obtained at 1.00 M urea-CaCl2. The highest shear strength of 728.5 kPa was determined at -2% OMC
when 1.00 M was used as urea-CaCl2 and 4.33% calcium carbonate content was yielded. It was determined that there is
a strong linear relationship (R2 = 0.8696) between UCS and calcium carbonate content in the treated soil. The amount of
calcium carbonate content formed was found to decrease with an increase in mixing water content from 2% dry to 4% wet of
optimum moisture content. Finally, the results from the field emission scanning electron microscope (FESEM) and energy
dispersive X-ray (EDX) analyses confirmed the formation of calcite crystals in the EICP treated soil.

Keywords Enzymatically-induced calcite precipitation (EICP) · Bio-cementation · Calcium carbonate precipitation ·

Residual soil

Introduction improvement techniques mimic nature in producing environ-

mentally friendly and sustainable materials for soil improve-
Bio-inspired and bio-mediated techniques have been ment [3–5]. The most studied aspect of bio-mediated soil
employed in soil improvement recently to reduce the nega- improvement technique is microbial induced calcite pre-
tive impact posed by global warming due to the production cipitation (MICP) via urea hydrolysis [6–13]. The MICP
of ­CO2, and also to mitigate the effects of toxic chemicals process utilizes either native or exogenous ureolytic bacteria
employed in soil stabilization and grouting [1, 2]. Unlike in conjunction with a cementation solution (urea and cal-
conventional methods of improvement, bio-mediated soil cium chloride) to produce calcium carbonate within the soil
matrix [1, 14–18]. Equations (1) to (4) summarise the major
* Muttaqa Uba Zango steps involved in MICP driven ureolysis.
[email protected] Urease enzyme
(1)
( )
NH2 2 CO + H2 O �����������������������→
� 2NH3 + CO2 [DIC]
1
School of Civil Engineering, Universiti Teknologi Malaysia,
81300 Johor Bahru, Johor, Malaysia
2
Department of Civil Engineering, KUST, 2NH3 + 2H2 O ↔ 2NH+4 + 2OH− (2)
Wudil P.M.B. 3244, Kano State, Nigeria
3
Department of Civil and Water Resources Engineering,
UNIMAID, Maiduguri P.M.B. 1069, Borno State, Nigeria

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78 Page 2 of 15 International Journal of Geosynthetics and Ground Engineering (2021) 7:78

CO2 + 2OH− ↔ HCO−3 + OH− ↔ CO2− + 2H2 O (3) concentration of cementation solution (urea-CaCl 2) at a
3
fixed amount of free urease enzyme (1 g/100 ml of water)
on the strength and hydraulic properties of sandy soil. The
Ca2 + CO2−
3
↔ CaCO3(s) (4) authors determined the highest UCS value to be at 1.0 M
urea-CaCl2. Other studies conducted on the treatment of
The central role of the microorganisms in MICP is to sandy soils with EICP include [22, 33–36].
secrete the urease enzyme, which will catalyze the above Critical studies into the findings of previous research
chemical reactions by increasing the pH of the environ- reveal conflicting results. Some authors determined that
ment and creating an initial supersaturated condition for efficient precipitation is achieved at lower concentrations of
the formation of calcite precipitates [19–21]. Despite urease enzyme (4 kU/L) and cementation solution (between
the promising potentialities of MICP, the process still 0.25 M and 0.5 M) [37, 38]. A few other researchers, such as
has some limitations to be regarded for field application. [39, 40] reported high calcium carbonate precipitations as
Among other challenges, as stated by [22], MICP through well as maximum UCS at high concentrations of urea-CaCl2
either injection or percolation of exogenous bacteria can- solution (1.5 M). Based on the precipitation test conducted,
not be applied in fine-grained soils such as silts and clays [22] recommended the optimum composition of EICP solu-
due to their smaller pore throats; the ureolytic activities of tion to be 3 g/L of urease enzyme, 0.67 M C ­ aCl2, 1.0 M urea,
the bacteria can also be subdued due to competition from and 4 g/L non-fat milk. Some researchers such as [37, 41]
other native bacteria, inadequate available oxygen, and soil have also studied the relationship between concentration of
pore fluid chemical composition. Even though, MICP were Urea-CaCl2 and urease enzyme concentration. For instance,
reported to have been applied in clay soils in few occa- [37] observed that the optimum concentration of urea-CaCl2
sions. For instance, [23–25] reported significant increase increased with the increment in the urease enzyme content.
in shear strength due to MICP application in various types [38] also reported similar observations. In this study, 3 g/L
of clay soils. Ting et al. [25] for example applied MICP via was adopted as the urease content based work of [22]. The
stepped injection method to improve shear strength of soils urease content adopted was expected to be sufficient to all
that was classified as gravelly clay of very high plasticity. the urea-CaCl2 chose for test tube test.
Sharma and Ramkrishnan [24] utilized MICP process via Most of the studies, as mentioned earlier, considered the
mix and compact method to increase shear strengths of improvement of sandy soils. However, based on a literature
clay soils of intermediate and high plasticity. search and to the best of our knowledge, EICP has never
The EICP technique serves as an alternative to MICP been applied to improve shear strength of fine-grained soils,
and solves many of the challenges associated with MICP especially residual clay soils. It is, therefore, imperative to
[26]. In EICP, free urease enzyme whose size is tiny conduct an experimental investigation to ascertain the opti-
(approximately 12 nm) and soluble in water, is used in mum concentration of EICP solution and also to consider the
place of ureolytic bacteria to catalyze the hydrolysis of application of the EICP technique in improving the mechani-
urea [27–30]. The free urease in EICP can be extracted cal properties of a fine-grained soil such as tropical residual
from agricultural produce, microbial or fungal sources. clay soil.
For instance, agricultural urease occurs in many high This work investigates the effect of varying concentra-
order plants such as sword bean, jack bean (Canavalia tions of urea-CaCl2 on the efficiency of calcite precipitation
ensiformis), soybean (Glycine max), melon seed (Citrullus (via the test tube test) and shear strength of the treated soil
lanatus), the pine family (Pinaceae), mulberry leaf (Morus through UCS test. The findings of this work could be of
alba), and pigweed (Chenopodium album) [2, 31]. More significance in understanding the effect of EICP treatment
so, the performance of urease enzyme sourced from plants on fine-grained soils.
or dead bacteria depend on the urease activity of the par-
ent sourced. Literature searched as summarised by [2] has
shown that even among plants, urease extracted from jack Materials and Methods
bean has the highest urease activity.
A few studies have been conducted on the EICP tech- Materials
nique for soil improvement. Nemati and Voordouw [32]
evaluated that an increase in the amount of urease enzyme The materials employed in this work are natural residual
from 0.01 to 0.1 g/L resulted in a higher rate of C ­ aCO3 soil and chemical reagents used for enzymatically-induced
formation, while the use of higher concentrations of calcium carbonate precipitation treatment.
cementation solution (urea and ­C aCl2) leads to the for-
mation of more qualitative calcium carbonate. Yasuhara
et al. [28] assessed the influence of varying the equimolar

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International Journal of Geosynthetics and Ground Engineering (2021) 7:78 Page 3 of 15 78

Table 1  Chemical reagents for Chemical reagent Source Property

EICP solution
Urease enzyme (Jack Bean) Fischer Scientific USA 90% Purity (3,500 U/mg)
Urea EMSURE® Malaysia A.R. 99% Purity
CaCl2.2H2O EMSURE® Malaysia A.R. 99% Purity

Natural Residual Soil plasticity based on the British Standard Classification Sys-
tem (BSC). The soil contains kaolinite as the dominant clay
The soil was collected at a depth of about 1.5 m from a sam- mineral. The natural residual soil has hydraulic conductiv-
pling area in Universiti Teknologi Malaysia, Johor Bahru ity value of 4.8 × ­10–7 m/s, and compaction parameters of
Campus ­(1o33′35’’ N, ­103o38′38’’E). The sampling area 1.48 Mg/m3 and 28%. The oxide contents analysis revealed
is characterized by tropical rainforest climatic condition that the ratio of silica-sesquioxide was 0.590; thus, the soil
with granite geologic formation [42]. The soil collected is is laterite soil [45].
reddish-brown, and the soil is dominated by kaolinite clay
minerals as revealed by an XRD analysis. Test Tube Test

EICP Solution The test tube test was conducted to determine the effect of
varying urea-calcium chloride concentrations on the mass
The EICP solution is composed of urea (CO NH2 2 ), cal- of calcium carbonate precipitations formed, ammonium ions
( )

cium chloride dihydrate (CaCl2 .2H2 O) , and free urease produced, and the pH of the solution. The EICP solutions
enzyme. The urea and calcium chloride dihydrate used are were prepared at different equimolar concentrations (0.25,
analytical grade reagents. The urease enzyme used to cata- 0.50, 0.75, 1.00, and 1.25 M) of urea-CaCl2. The solutions
lyze the formation of calcium carbonate is a high purity, free were made by dissolving urea and calcium chloride dihy-
urease sourced from Canavalia ensiformis (jack bean). The drate in distilled water. Free urease enzyme sourced from
urease enzyme was manufactured by Fischer-Scientific USA jack bean with 3500 U/g activity was added (at 3 g/l) imme-
and purchased from Fischer-Scientific Sdn Bhd, Malaysia. diately after dissolution of urea-CaCl2, and then the EICP
Table 1 gives summary of the information about the chemi- solution was mixed for about 3–5 min. For each urea-CaCl2
cal reagents used for EICP treatment. concentration, a triplicate of 20 ml of EICP solutions were

Methods Table 2  Index properties of the natural soil

Property Quantity
Index Properties and Compaction Test
Natural moisture content (%) 32.72
To characterize the natural residual soil, laboratory tests that Percentage passing 63 μm sieve (%) 58.00
include particle size analysis, Atterberg limits tests (liquid Gravel fraction (%) 16
limit, plastic limit, plasticity index and linear shrinkage), and Sand fraction (%) 26
specific gravity were performed in accordance to [43]. The Silt fraction (%) 40.5
liquid limit test was conducted using the cone penetrometer Clay fraction (%) 17.5
equipment on the sample that passed through a 425 μm. The Liquid limit (%) 79
specific gravity was performed on a soil sample passing a Plastic limit (%) 30
2 mm sieve using the small pycnometer test. British Stand- Plasticity index (%) 49
ard Light (BSL) as described in [44] was followed in this Linear shrinkage (%) 16
study to compact the soil in order to determine optimum Specific gravity 2.63
moisture content and maximum dry density. Loss on ignition (LOI) (%) 12.28
The results of the index properties and basic engineer- BSCS classification MV (Silty Sand of
Very High Plastic-
ing properties tests on the natural residual soil are summa- ity)
rised in Table 2. The particle size distribution of soil shows Colour Reddish Brown
that the soil consists of 16% gravels, 26% sands, 40.5% silts Clay minerals Kaolinite
and 17.5% clays. The Atterberg limits results indicate that Maximum dry density (MDD) (Mg/m3) 1.48
the soil is high plastic with 79% LL, 30 PL and 49% PI. Optimum moisture content (OMC) (%) 28
Therefore, the soil was classified as sandy silts of very high Permeability (m/s) 4.8 × ­10–7

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78 Page 4 of 15 International Journal of Geosynthetics and Ground Engineering (2021) 7:78

prepared to ensure reproducibility. The pH of the urea-CaCl2 to 26%, while the 4% is equivalent to 32%. The cementation
solution was measured before and after adding the urease solutions were first prepared by dissolving equimolar con-
enzyme. The prepared solutions were incubated at 30 °C centration of urea and calcium chloride in distilled water.
for 3, 7, and 14 days. At the end of each incubation age, the Then free urease enzyme at 3 g/L concentration was added
mass of precipitations formed was determined by passing the immediately to the dissolved urea-CaCl2 solutions. The urea-
EICP solutions through a filter paper until the filtered liquid CaCl2 and urease enzyme mixture was mixed thoroughly
stopped dripping. The white precipitates on the filter paper until all the enzyme were dissolved.
and in the tubes were dried for 24 h in an oven operating at The mix and compact method of treatment was used in
50 °C. The actual mass of precipitations was determined preparing UCS samples in this study, as reported by [22, 29,
by combining the mass of precipitation residue on the filter 46]. The prepared EICP solutions were mixed with dry soil
and in the tubes. that passed through 2 mm sieve and then allowed moisture to
Ammonium ions from the filtered liquid were determined equilibrate. The soil–EICP solutions mixed were compacted
by following the procedure described in the Nessler method, in mould (38 mm diameter by 76 mm height) using static
while the change in pH of the solutions was determined compaction machine as shown in Fig. 2. During preparation,
using a pH meter. Finally, the mineral composition of the the soil was poured into the mould in three layers, each layer
precipitation formed from the test tube test was analyzed by was tampered 15 times with the fixed mass of cylindrical
performing the XRD analysis. Figure 1 shows the schematic rod before compressed with static compaction machine. The
diagram of the procedure followed for conducting test tube samples were compressed in two stages from the top and
test. bottom by using two platens of different heights. During the
second staged compression, the platens were interchanged
EICP Treatment and Unconfined Compressive Strength Test and also the mould’s position was changed upside down.
Note that the compactness of the samples were measured
The EICP solutions were prepared at various concentrations based on the predetermined mass of dry soil (corresponding
of urea-calcium chloride (0.25, 0.5, 0.75 and 1.0 M) and dif- to dry density) required to fill up the fixed volume of UCS
ferent moulding water contents ranging from 2% dry to 4% mould (36 mm diameter and 76 mm height). The compacted
wet relative to optimum moisture content, which is 28%. The samples were cured for 3 days inside humidity chamber
water content corresponding to 2% dry of OMC is equivalent operating at 30 °C.

Fig. 1  Schematic procedure of test tube test

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International Journal of Geosynthetics and Ground Engineering (2021) 7:78 Page 5 of 15 78

Fig. 2  Schematic procedure of EICP treatment for UCS Test

The cured samples were subjected to unconfined com- The microstructures of the untreated residual and EICP-
pressive strength. The unconfined compressive strength treated soils were studied by conducting FESEM and EDX
(UCS) test was conducted by following the procedure analyses. The tests were conducted in a SU8020 Hitachi
described in [47]. The UCS test was performed by mount- machine at UTM’s central laboratory.
ing untreated and EICP treated UCS samples on a triaxial
machine. The samples were then subjected to uniaxial
compression at a rate of 1.52 mm/min until failure, and Results and Discussion
the displacement with corresponding forces were auto-
matically recorded in a computer using DS 7 software. Results on Test Tube Test

Assessment of environmental factors such as concentration

Determination of Calcium Carbonate Contents of cementation solution, temperature and pH, are prerequi-
and Microstructure Analysis site to full field implementation of MICP process [50]. Test
tube test has been used to study the effect of various factors
The samples that were subjected to the UCS test were such as concentration of cementation solution, temperature
then used for the determination of calcium carbonate and pH on the efficiency of EICP and MICP techniques.
content by conducting acid digestion test. About 20 g Various authors such as [26, 37, 51] had adopted the use of
of oven-dried soil samples were collected from the top, test tube test to study factors such as molarity of cementa-
middle and bottom part of each EICP-treated sample for tion reagents, and concentration of urease enzyme on the
calcium carbonate content determination. The calcium efficiency of EICP process.
carbonate contents were determined by adapting the The test tube test was conducted to establish the con-
gravimetric acid wash method, as described by [48, 49]. cept of urea hydrolysis by employing free urease enzyme.
The dried samples were dissolved in HCl acid at a con- The results of the test were evaluated to determine the opti-
centration of 4 M. The dissolved calcium carbonate, soil mum EICP solutions from the various concentrations of
and acid mixtures were rinsed multiple times with water, urea-CaCl2.
and passed through a 63 μm sieve and filtered paper. The
retained soils on the sieve were weighed after oven dry- Effect of Cementation Reagents on the Calcium Carbonate
ing. The difference in weight between the dried sample Precipitates
before acid digestion and the dried sample post-acid
washing is regarded as the mass of calcium carbonate. Figure 3 illustrates the effect of varying urea-CaCl2 con-
The formula for calculating calcium carbonate content is centrations on the mass of calcium carbonate precipitation
expressed in Eq. (5) below: at three different incubation periods (3, 7 and 14 days).
Regardless of the incubation period, there is a general
B
( )
CCC = 100 − × 100 (5) increase in the mass of precipitations with higher urea-
A
CaCl 2 concentrations from 0.25 until 1.00 M; the mass
CCC = Calcium carbonate content (%). of precipitations decreases with a further increase in the
B = Mass of oven dried soil post washing. concentration of cementation reagents to 1.25 M. The
A = Mass of oven-dried treated soil before washing. maximum amount of precipitations occurs at 1.00 M
urea-CaCl 2. The increase in the mass of precipitation

13
78 Page 6 of 15 International Journal of Geosynthetics and Ground Engineering (2021) 7:78

Fig. 3  Correlation between 1.80

urea-CaCl2 and mass of cal-
cium carbonate precipitation at 1.60 3 Days
various curing age 7 Days
1.40
14 days

Precipitation Mass (g)

1.20

1.00

0.80

0.60

0.40

0.20

0.00
0.25 0.50 0.75 1.00 1.25
Urea-CaCl2 Conc. (M)

with an increase in cementation reagent (urea-CaCl 2) Conversely, the lower precipitation ratio in higher concentra-
concentration is due to the high amount of urea available tions of urea-CaCl2 reagent (1.25 M) is attributed to insuf-
for conversion to ammonia and carbonate ions and also ficient urease enzyme content and the suppression action of
a high amount of calcium ions ­( Ca 2+) [52]. The EICP high concentration cementation reagent on enzyme activity
recipe obtained in this study (3 g/l urease enzyme; 1 M [28, 29]. Unlike the results obtained by [29, 38], this study
urea-CaCl2) is similar to the one obtained by [22], that finds that precipitation ratio increases with an increase in the
is 1.00 M and 3 g/L in terms of urea concentration and concentration of urea and calcium chloride.
urease amount, respectively, but differs in respect of the
Precipitation ratioPR(%)
concentration of ­CaCl2.
The results of the test tube test can also be expressed = Actual mass of CaCO3 ∕Theoretical mass of CaCO3
in terms of precipitation ratio, which is the measure of the (6)
process efficiency. The formula used to compute the pre- where Theoretical Mass ofCaCO3 = C × V × M
cipitation ratio is expressed in Eq. (6) was adopted from C: concentration of urea-CaCl2 in molarity.
the works of [22, 26, 51]. As shown in Table 3, the highest V: volume of urea-CaCl2 solution used in litre.
efficiencies of at least 90% were obtained in 0.5 and 0.75 M M: molar mass of ­CaCO3 in g/mol.
urea-CaCl2 concentrations at all the incubation periods of Figure 3 also shows that there is no significant increase in
3, 7, and 14 days. Precipitation ratios of more than 80% the mass of calcium carbonate precipitation when the EICP
were obtained for 0.25 M urea-CaCl2 concentrations while solutions were incubated beyond three days. Therefore,
approximately 80% efficiencies were obtained for the it can be concluded that urea hydrolysis is accomplished
1.00 M cementation reagent when the solution was cured within a short time of not more than three days. Similar
for between 3 and 14 days. The lowest efficiency of at most results were reported by [53], whereby they determined that
40% was recorded for the 1.25 M urea-CaCl2 concentration. the increase in the amount of calcium carbonate precipita-
These findings indicate that the amount of urease enzyme tion is not affected greatly by curing period.
(3 g/L) adopted for catalyzing the reaction was enough for The result of XRD analysis on the precipitates, as shown
urea-CaCl2 concentrations of 0.25, 0.50, 0.75 and 1.00 M. in Fig. 4 has confirmed the presence of calcite and vaterite.
These are two crystalline forms of calcium carbonates. It is
Table 3  Effect of cementation reagents on precipitation ratio
seen that about 81.6% of the minerals from the precipitation
Urea-CaCl2 Conc. Precipitation ratio (%) were calcite and 18.4% vaterite. Thus, the treatment of the
(M)
3 days 7 days 14 days residual via EICP is expected to be stable, as calcite is more
stable than vaterite. The same minerals were obtained when
0.25 81.93 82.56 85.26
test tube test experiments were conducted by [37, 54, 55].
0.50 89.92 90.26 90.59
Wen et al. [56] found that calcite polymorphs were the domi-
0.75 95.25 96.58 97.02
nant mineral formed due to EICP, while vaterite polymorphs
1.00 78.77 80.10 80.10
were the dominant mineral formed if the process was MICP.
1.25 40.23 40.96 41.96

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International Journal of Geosynthetics and Ground Engineering (2021) 7:78 Page 7 of 15 78

Fig. 4  XRD analysis of precipi-

tations from test-tube test

Ammonium Content and pH values 3 days after urease enzyme was added to the cementa-
­ H4+ reduced
tion solutions, after that, the concentration of N
The EICP and MICP processes involve the decomposition at 7 and 14 days.
of urea into two moles ammonium and one-mole carbon- As also shown in Fig. 5, 0.75 M EICP solution has the
ate ions. Presence of ammonium ions and increase in pH lowest concentration of ammonium ions at all the curing
are two indicators used to study the rate urea hydrolysis in periods, while 1.25 M EICP solution has the highest ammo-
either the MICP or EICP processes [57, 58]. Figure 5 shows nium ions concentration. This indicates that the rate of urea
the variation of ammonium contents with curing periods for decomposition is faster in 0.75 M EICP solution than in the
the various concentrations of cementation solutions. Before remaining EICP solutions [58]. Thus, 0.75 M EICP solution
the addition of the urease enzyme, ammonium ion content remained the best in terms of precipitation efficiency and
was found to be zero in all the concentrations of cementa- rate of urea decomposition.
tion solutions. The ammonium ions attained their maximum

Fig. 5  Change in ammonium 1.80

content with curing ages
1.60 0.25 M

0.50 M
1.40
0.75 M
1.20
NH4+ Conc. (mg/l)

1.00 M
1.00
1.25 M
0.80

0.60

0.40

0.20

0.00

-0.20
3 7 14
Curing Age (Days)

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78 Page 8 of 15 International Journal of Geosynthetics and Ground Engineering (2021) 7:78

The rate of formation of calcium carbonate precipitation Unconfined Compressive Strength

was not investigated in this study, therefore, the exact time
at which precipitation started was not measured. However, Unconfined Compressive Strength Versus Concentration
it was observed that the EICP solution becomes turbid a of Cementation Reagent
few hours after addition of urease enzyme. Visible precipi-
tations were clearly seen at the bottom of the test tube after The effect of calcium carbonate precipitation on the shear
1–2 days. Similar observations were reported by [59]. Thus, strength of EICP-treated residual soil was assessed by per-
the quantification of precipitations in this study was started forming the unconfined compressive strength test. Figure 7
at 3 day based on the work of [41]. It is expected that the presents the variations in UCS with a change in urea-CaCl2
earliest time at which ureolytic reaction could be completed concentration at various moulding water contents. The
is 72 h. graph shows a general rise in compressive strength with an
Figure 6 shows the variation of pH values of various increase in urea-CaCl2 concentration. Peak strength values
concentrations of urea-CaCl2 (Cementation) solutions with were obtained at 1.00 M cementation reagent. The increase
curing time. It can be seen from the figure that the pH of in UCS is due to the formation of calcium carbonate pre-
urea-CaCl2 (before urease enzyme were added) was slightly cipitation which increases the existing bonding between soil
acidic, and they fall within the range of 6.02 – 6.75. These particles [63]. In this study, the higher UCS values were
values were similar to pH reported by [58, 60]. The pH obtained at higher cementation solutions. For example, the
of the various EICP solutions spiked to maximum values highest recorded strength of 728.5 kPa was determined to
(between 8.52 and 8.02) within five minutes after the addi- be at 1.00 M cementation solution when the soil was pre-
tion of the urease enzyme. The pH values dropped at three pared at -2% OMC. The pattern of results determined in this
days of curing time, and further reduced gradually at 7 and study is similar to some previously published studies like
14 days. As explained by [60], the sudden increased in pH [52, 64–67]. For instance, [64] determined that the UCS
after addition of urease enzyme is attributed to the rapid value of aeolian sand increases from 1.71 MPa to 18 MPa
decomposition of urea substrate into ammonium ions and with an increase in the concentration of cementation solu-
hydroxyl ions and it continues to increase until the EICP tion from 0.5 M to 2.5 M. Deng and Wang [65] reported a
solutions were supersaturated. Upon precipitations of cal- maximum unconfined compressive strength of 2.61 Mpa at
cium carbonate (at day 3), the pH values dropped as stated a 1 M corresponding concentration of urea-CaCl2. Similarly,
by [61]. The pH values continued decreasing (beyond 3 days [68] found that the highest UCS of 0.81 MPa was produced
until 14 days) until all the calcium ions were converted to when sandy soil was treated with 1.5 M urea-CaCl2 via the
­CaCO3 [58]. A similar pattern of results was reported by EICP technique.
many researchers such as [57, 58, 60, 62]. For example, [58]
found that the initial pH values of EICP solution raised from Effect of EICP Treatment on the Failure Mechanism
6.6–6.8 to 8.5–8.8 due to addition of urease enzyme. The pH
values were then dropped to 8 after the solution had reached Figure 8 shows the stress strain curves of untreated and
supersaturation level. EICP treated soils prepared at 0% OMC moulding water

Fig. 6  Change in pH with cur- 9.00 Urea-CaCl2 (No

ing age
8.00 urease)
3 Days
7.00
7 Days
6.00
5.00 14 Days
pH

4.00
3.00
2.00
1.00
0.00
0.25 0.50 0.75 1.00 1.25
Conc. of Urea-CaCl2 (M)

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International Journal of Geosynthetics and Ground Engineering (2021) 7:78 Page 9 of 15 78

Fig. 7  Variation of unconfined -2%OMC

compressive strength with 800
urea-CaCl2 at various moulding
0%OMC
water contents

600 +2%OMC

UCS (kPa)
+4%OMC

400

200

0
Natural 0.25 0.50 0.75 1.00
Urea-CaCl2 Conc. (M)

800 content. The impact of EICP treatment on the failure

700
mechanism were clearly seen in the figure. As shown in
0.25 M
the figure, it can be seen that there is general increase in
0.50 M
600
0.75 M shear strength and general decreased in shear strain at
500 1.00 M failure upon EICP treatment and with further increase in
Stress (kPa)

Untreated
the concentration of cementation solution. For instance,
400
the maximum shear stress and shear strain at failure of
300 untreated soil are 264 kPa and 3.21%, respectively, with
200 EICP treatment at 1.00 M cementation solution, the
maximum shear stress and axial strain were 710 kPa and
100
2.79%, respectively. It can be seen that the EICP treat-
0 ment generally resulted in maximum shear strength at
0 1 2 3 4 5 6
low strain. It can be clearly seen that the failure mode
Axial Strain (%)
of untreated soil changed from ductile to brittle failure
upon EICP treatment and with increase in cementation
Fig. 8  Stress strain curves of untreated and EICP treated soils pre-
pared at 0% OMC moulding water content solutions concentrations. Figure 9 also clearly explain the

Fig. 9  Images showing failure various patterns in (a) Untreated, (b) 0.25 M, (c) 0.50 M, (d) 0.75 M and (e) 1.00 M EICP treated soils

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failure pattern of the untreated and EICP treated soils. Influence of Water Content and Cementation Reagents
Similar pattern of stress – strain curve was reported by on Calcium Carbonate Content
[69] when MICP were employed for the treatment of
sandy soil. The effect of cementation solution concentrations and
moulding water contents are explained in Fig. 11. Firstly, it
can be seen that the amount of calcium carbonate increases
Unconfined Compressive Strength Versus Calcium with the increase in the concentration of cementation rea-
Carbonate Precipitates Content in Treated Soil gents. By way of illustration, the calcium carbonate content
of samples prepared at 0% OMC rises from 1.64% at 0.25 M
The relationship between the unconfined compressive to 4.18% at 1.00 M. The increase in calcium carbonate con-
strength versus calcium carbonate content formed within the tent with urea-CaCl2 can be attributed to the high amount
EICP-treated soil is shown in Fig. 10. The graph shows an of urea to be converted and also a high concentration of
increase in UCS values with an increase in calcium carbon- available calcium ions [52]. The same pattern of results was
ate precipitation. For instance, there is an increase in UCS reported by [65, 70].
from 498 kPa to 728.5 kPa due to an increase in calcium The influence of moulding water content within the
carbonate content from 1.05% to 4.33%. The same pattern concentration of cementation solution is also depicted in
of results was reported by [35, 66, 68]. Fig. 11. The calcium carbonate content decreases with an

Fig. 10  Relationship between 1000

UCS and calcium carbonates 0% OMC
contents
900 -2% OMC
+2% OMC
800 R² = 0.8696 +4% OMC
UCS (kPa)

700

600

500

400
0 1 2 3 4 5
Calcium Carbonate Content (%)

Fig. 11  Variation of calcium 5.0 -2% OMC

carbonate content with urea-
Calcium Carbonate Content (%)

CaCl2 at various moulding

water contents
0% OMC
4.0
+2% OMC
3.0 +4% OMC

2.0

1.0

0.0
0.25 0.50 0.75 1.00
Urea-CaCl2 Conc. (M)

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International Journal of Geosynthetics and Ground Engineering (2021) 7:78 Page 11 of 15 78

increment in moulding water content from -2% to + 4% water contents ( – 2%, 0%, + 2% and + 4%) and urea-CaCl2
OMC. For example, in samples treated with 1.00 M cemen- concentrations for UCS test. Calcium carbonate content in
tation solution, the calcium carbonate content decreases the EICP treated soils were determined by conducting acid
from 4.333% to 2.233% with an increase in moulding water digestion while the mineral content in the treated soil was
content from 2% dry to 4% dry relative to optimum moisture investigated via FESEM and EDX analysis. Based on the
content. The reason for the formation of more calcium car- tests conducted and results obtained the following conclu-
bonate precipitates in the samples prepared at the negative sions were made:
side of the optimum is attributed to the absorption of more
cementation reagents due to the less saturated soil particles I. Amount of calcium carbonate precipitation increases
[71, 72]. A similar pattern of results was reported by [73]. with an increase in the concentration of cementation
reagent. The highest calcium carbonate precipitation
Microstructural Analysis of the EICP Treated Soil of 1.603 g was obtained at 1.00 M urea-CaCl2 when
the solution were cured at 30 °C for 7 and 14 days. It
The formation of calcium carbonate precipitation was exam- was also found that curing period beyond 3 days has
ined through FESEM and EDX analyses of both natural and insignificant effect on the amount of calcium carbon-
EICP-treated soils (Figs. 12 and 13, respectively). As shown ate precipitation formed.
in Fig. 12(b), calcite precipitates can be seen sparsely depos- II. The XRD analysis performed on the precipitations
ited on the treated soil. An EDX analysis conducted on the formed from the test tube test revealed the forma-
natural soil (see Fig. 13a) and EICP-treated soil (shown in tion of calcite crystals. Thus, EICP treatment of the
Fig. 12b), confirmed the presence of calcium and carbon ele- residual is expected to be stable.
ments. It can be seen that the percentage of Ca and C in the III. The strength of the treated soil rises with higher
untreated samples were zero, as they are not detected, while cementation solution concentrations and calcium
the percentages of these elements were 1.7 and 17.5% in the carbonate content. The maximum UCS of 728.5 kPa
EICP treated soils. The Ca and C are parts of the elements was determined to be at 1.00 M urea-CaCl 2 and
that make up ­CaCO3. 4.33% calcium carbonate content. It was also deter-
mined that the shear strength in the EICP treated soils
increase with increase in the amount of calcium car-
Conclusions bonate content.
IV. The shear strength as well as calcium carbonate pre-
Bio-mediated soil improvement via the use of free urease cipitation was seen to be influenced by the moulding
enzyme was employed to enhance the shear strength of water content used to prepare the soil samples. Irre-
residual fine grained soil. The optimum concentration of spective of molarity of cementation solution, highest
cementation reagent (urea-CaCl2) was determined by con- shear strength and calcium carbonate precipitations
ducting a test tube test. The mix and compact method was were determined at moulding water content of 2 dry
adopted in preparing EICP-treated soil at varying moulding of optimum moisture content.

Fig. 12  FESEM image of a

Natural residual Soil and b
treated residual soil at X30.0 K

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78 Page 12 of 15 International Journal of Geosynthetics and Ground Engineering (2021) 7:78

Fig. 13  EDX analysis on a natu-

ral and b EICP treated soil

V. The presence of calcite crystals in the treated soil II. The results of this study would be beneficial in
was confirmed through FESEM and EDX analyses the improvement of marginal soil for the design of
in the EICP treated soil. Ca and C elements were earthen facilities such as compacted clay liner, road
not detected in the untreated soils, while 1.7% and subgrade and rammed earth structures.
17.5% of Ca and C, respectively were determined in
the EICP treated soils.
Acknowledgements The authors gratefully acknowledge the finan-
cial support from the Malaysian Ministry of Higher Education
The following recommendations are made: (MOHE) through the Fundamental Research Grant Scheme (FRGS):
(R.J130000.7851.5F256), GUP from UTM: Q.J130000.2522.20H21
I. The effect of environmental factors, such as curing and High Impact Research Q.J130000.2451.04G57 from UTM. The
temperature and treatment cycles, should be studied.

13
International Journal of Geosynthetics and Ground Engineering (2021) 7:78 Page 13 of 15 78

first and last authors also acknowledge the funding coming from the 13. Xiao Y, Asce M, Wang Y, Desai CS, Asce DM (2019) Strength
Nigerian Government through TETFUND. and deformation responses of biocemented sands using a tem-
perature-controlled method. Int J Geomech 19:1–10. https://
Author Contributions The author Muttaqa Uba Zango developed ascelibrary.org/doi/abs/https://​d oi.​o rg/​1 0.​1 061/%​2 8ASCE%​
the experimental work, analysed the results and prepared the manu- 29GM.​1943-​5622.​00014​97
script. The authors Khairul Anuar Kassim conceived the concept of 14. Terzis D, Laloui L (2019) A decade of progress and turning
idea biocementation via EICP on residual soil. The author Kamarudin points in the understanding of bio-improved soils: a review.
Ahmad verified the experimental procedure and review the manuscript. Geomech Energy Environ. https://​doi.​org/​10.​1016/j.​gete.​2019.​
The author Abubakar Sadiq Muhammed performed the experimental 03.​001
works. 15. Gomez MG, Anderson CM, Graddy CMR, DeJong JT, Nelson
DC, Ginn TR (2017) Large-scale comparison of bioaugmenta-
tion and biostimulation approaches for biocementation of sands. J
Geotech Geoenvironm Eng 143:04016124. http://ascelibrary.org/
doi/https://​doi.​org/​10.​1061/%​28ASCE%​29GT.​1943-​5606.​00016​
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