Geomicrobiology Journal

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Microbially Induced Calcite Precipitation for

Seepage Control in Sandy Soil

Yufeng Gao, Xinyi Tang, Jian Chu & Jia He

To cite this article: Yufeng Gao, Xinyi Tang, Jian Chu & Jia He (2019): Microbially Induced
Calcite Precipitation for Seepage Control in Sandy Soil, Geomicrobiology Journal, DOI:
10.1080/01490451.2018.1556750

To link to this article: https://doi.org/10.1080/01490451.2018.1556750

Published online: 24 Jan 2019.

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GEOMICROBIOLOGY JOURNAL
https://doi.org/10.1080/01490451.2018.1556750

Microbially Induced Calcite Precipitation for Seepage Control in Sandy Soil

Yufeng Gaoa,b, Xinyi Tanga,b, Jian Chuc, and Jia Hea,b
a
Key Laboratory of Ministry of Education for Geomechanics and Embankment Engineering, Hohai University, Nanjing, China;
b
Jiangsu Research Center for Geotechnical Engineering Technology, Hohai University, Nanjing, China; cSchool of Civil and Environmental
Engineering, Nanyang Technological University, Singapore

ABSTRACT ARTICLE HISTORY

Microbially induced calcite precipitation (MICP) can reduce the permeability of soil by reducing Received 14 July 2017
the pore volumes. A MICP-based soil improvement method to control water leakage in irrigation Accepted 3 December 2018
channels and reservoirs built on sandy soil grounds is presented in this article. Using this method,
KEYWORDS
a low-permeable hard crust can be formed at the soil surfaces. An experimental study was carried
Microbially induced calcite
out to evaluate the effect of this method. Sandy soil samples were treated using four different precipitation; permeabil-
schemes, namely, (1) surface spray, (2) surface spray with the addition of fibers, (3) surface spray ity; ureolysis
and bulk stabilization, and (4) immersion stabilization. By applying around 2.6 L treatment liquid
(consisting of ureolytic bacteria, 0.5 mol/L calcium chloride and 0.5 mol/L urea) to the top 2-cm
thick soil, the seepage rates of the samples treated by the four different schemes could be
reduced by up to 379 times. The conversion rates of calcium source in the tests were up to
89.7%. The results showed that a method of treating the soil in bulk before the formation of a
crust on top of the soil layer was effective in reducing the seepage rates. After the bio-treatment,
the formed low-permeable hard crust layer was 10 to 20 mm thick with a calcite content higher
than 5%. Below the hard crusts, the calcite content was less than 5% and the soil was not prop-
erly cemented. Using the mercury intrusion test, it was found that both pore volumes and pore
sizes of the bio-treated soil reduced significantly as compared with the untreated soil. Penetration
tests using a flat-bottom penetrometer were used to assess the mechanical behavior of the bio-
treated soil. The results indicated that the penetration resistance of the bio-treated soil layer was
much higher than that of the untreated soil.

Introduction to be used in large scale. New materials and methods have

In arid regions in China, efficient utilization of water
resources is still a challenge. Many reservoirs and channels
are built in wind-formed sandy soils. This type of soil usu-
ally consists of almost pure sand particles. As a result, the
permeability of the ground soil is high and the leakage prob-
lem is severe. In China, around 80% irrigation channels do
not have any leakage control measures (Li 2000). More than
half the water resources for agricultural use are wasted in
the process of transportation. In this condition, solutions are
required for the control of water leakage in sandy
soil ground.
Conventional engineering measures for the control of
leakage involves compacted clay liners, bentonite liners, geo-
membranes, concrete covers, polymer coatings, etc. Clay lin-
ers and bentonite liners have very low permeability. The
typical permeability values range from 10
4 to 10
10 cm/s
(Dong et al. 2015). Cracks can develop in clay and
bentonite due to volume shrinkage in dry conditions.
Impermeable concretes have 10
7–10
12 cm/s permeability.
Geomembranes have 10
11–10
13 cm/s permeability.
However, these two types of materials are relatively expensive
been tested for the leakage control in soils, such as extracellu-
lar polymeric substances (EPS) (Blauw et al. 2009; Lentz
2007; Proto et al. 2016; Ragusa et al. 1994; Tang et al. 2018a,
2018b). The production of EPS in soils can significantly
reduce the permeability of soils by clogging soil pores and
increase the viscosity of pore water. However, EPS can be
degraded by some other microbes, and EPS does not contrib-
ute greatly to the strength of soils. So, the stability of soil
properties treated by EPS is questionable. Besides, this new
method is still in development and has not yet been used in
large-scale applications.
Microbial mineral precipitation processes are widespread
in nature. Calcite, the stable form of calcium carbonate, can
be produced by several microbial reactions, such as micro-
bial ureolysis and microbial denitrification reactions (Al
Qabany and Soga 2013; Li et al. 2017; Van Paassen et al.
2010; Whiffin et al. 2007). This process is named microbially
induced calcite precipitation (MICP). Ureolysis-based MICP
as a soil modification method for increasing the strength or
reducing the permeability has been adopted in most related
studies (DeJong et al. 2013). In this method, urea is hydro-
lyzed into ammonium and carbonate by the ureolytic

CONTACT Jia He hejia@hhu.edu.cn Key Laboratory of Ministry of Education for Geomechanics and Embankment Engineering, Hohai University, Nanjing
210098, China
ß 2019 Informa UK Limited, trading as Taylor & Francis Group
2 Y. GAO ET AL.

bacteria. The hydrolyzation of urea also leads to the net

increase of pH due to the ionic equilibrium (Mobley and
Hausinger 1989). If aqueous calcium is present in the sys-
tem, calcite can be precipitated. The reaction is as follows:
ureolytic bacteria
NH2
CO
NH2 þ 2H2 O
! 2NH4 þ þ CO3 2
[1]

Ca2þ þ CO3 2
! CaCO3 # [2]

Bacteria have a strong tendency to attach to surfaces of
soil particles. When calcite is formed around the bacterial
cells, it can fill soil pores, bridge soil particles, and in turn
reduce the permeability and improve the strength of soils.
In recent years, the MICP process has been extensively
studied for different applications in civil engineering and
constructions. The MICP process can result in a persistent
reduction in soil permeability. This method can be used for
the construction of reservoirs (Chu et al. 2012, 2013; Figure 1. Photograph of the soil particles.
Stabnikov et al. 2011), the control of water flow-induced soil
erosion (Jiang et al. 2017; Jiang and Soga 2017), and the for- soil particles is presented in Figure 1. The result of the XRD
mation of barrier to control contaminant migration analysis on the soil minerals is given in Figure 2. It shows
(Cuthbert et al. 2013; Fujita et al. 2010; Li et al. 2013). As that the soil mainly consists of quartz. The particle size dis-
for seepage control in soils, the MICP treatment can simply tribution is shown in Figure 3. It is a typical aeolian soil in
be carried out by applying the bacteria and treatment mate-
Northwestern China. In many arid and desertified regions
rials onto soil surfaces (Al Qabany et al. 2012; Chu et al.
in China, surface soils are composed of almost pure
2012, 2013; Liu et al. 2016; Stabnikov et al. 2011). A low-
sand particles.
permeable crust layer can be formed at the soil surfaces and
The test set-up is presented in Figure 4. Soil was
impede the water infiltration from soil surfaces. The
placed in plastic buckets for the microbial treatment and
strength of soil surfaces can also be improved to resist
testing. The diameter of the buckets was 18.5 cm. A 2
mechanical loads and other environmental impacts. The
cm-thick gravel layer was placed on the bottom of the
MICP method is ideal to control water leakage and enhance
the efficient utilization of water resources in arid regions buckets. Dry sandy soil was placed into the buckets and
(Chu et al. 2012, 2013; Gollapudi et al. 1995). As compared compacted to achieve a dry density of 1.46 g/cm3. There
with other engineering methods for seepage control, the was a metal filter between gravel and soil. Two holes
MICP method can be more cost-effective and environmen- were drilled on the bottom of the bucket for the connec-
tally-friendly. The bacterial species used for the MICP treat- tion of tubes for the microbial treatments and the seepage
ment such as Sporosarcina pasteurii are usually isolated rate tests.
from natural soil environments. Recycled materials from
agricultural and domestic wastes can be utilized for the bac-
terial cultivation and the MICP treatment (Cheng and Cord- Microbial culture
Ruwisch 2013; Choi et al. 2016). Sporosarcina pasteurii (CGMCC1.3687) from China General
In this paper, an experimental study was carried out to Microbiological Culture Collection Center was used in this
investigate the performance and mechanism of the MICP study. According to the specification, this species is the
for the control of water leakage in sandy soil. Four different same one as ATCC 11859. The bacteria were cultivated in
treatment schemes, namely, surface spray, surface spray with the liquid medium consisting of: yeast extract 20 g/L, NH4Cl
fibers, surface spray and bulk stabilization, and immersion 10 g/L, NiCl26H20 2.4g/L, MnSO4H2O 1g/L, and NaOH
stabilization, were tested. Permeability tests were conducted solution for the adjustment of pH to 8.5. The liquid medium
to evaluate the effect of seepage control. Penetration tests
was autoclaved before inoculation. The bacteria were culti-
were conducted to measure the surface strength after the
vated aerobically at 30  C and at 100 rpm shaking in an
treatment. The micro-scale pore size characteristics of the
incubator for 24 h before harvest. Harvested cells were cen-
bio-treated soil were studied using a mercury intrusion
trifuged and dispersed in 9 g/L NaCl solution for storage
porosimeter.
at 4 C.
The activity of bacteria was determined by an electrical
Materials and methods conductivity method. The ureolysis reaction involves the
hydrolysis of non-ionic substrate urea to ionic products,
Soil and experimental set-up
leading to the proportional increase in the electrical con-
Soil used in this study was obtained from Wuwei, Gansu ductivity. The bacteria used in this study had activities from
Province, located in Northwest China. A photograph of the 0.3 to 0.5 mS/cm/min, or 2.65–4.42 mM/min.
 GEOMICROBIOLOGY JOURNAL 3

Figure 2. Result of XRD analysis of soil minerals.

Water/Treatment liquid
The hard crust

Sand
Metal filter
Gravel

Container
Figure 3. Particle size distribution of soil.
Balance
Figure 4. Schematic diagram of soil treatment and the seepage rate
test apparatus.
Microbial treatment
The treatment liquid was obtained by mixing bacterial liquid time interval between two consecutive treatments was
and urea-calcium chloride solutions together at 1:1 volume 2 d.
ratio. The treatment liquid contained 0.5 mol/L equimolar
(2) Surface spray with the addition of fibers
urea and calcium chloride. The treatment liquid was applied
Before the application of treatment liquids, polypropylene
to soil samples by four different schemes as
fibers were evenly placed on the surface of soil samples at
explained below.
37.3 g/m2. The size of the fibers was around 10 mm in
(1) Surface spray length and 0.1 mm in diameter. The amount of treatment
Treatment liquids were applied onto the surface of soil liquid applied to soil and the application method were
samples by spraying using a spray pot. The volume of the same as those in scheme (1).
treatment liquids applied each time was 134 mL (5 L/
(3) Surface spray and bulk stabilization
m2). Total 19 times spray treatments were applied. The
4 Y. GAO ET AL.
Table 1. Treatment methods and brief results.
No of bulk No of immersion
Treatment schemes stabilization stabilization No of surface spray
I 0 0 19
II 0 0
III 3 0
IV 0 5
Prior to the spray treatments, soil samples were immersed
in the treatment liquid for the bulk stabilization of soil
for three times. The treatment liquid retained at the soil
surface for 48 h. After the bulk stabilizations, the soil
sample received spray treatments for 12 times using the
same method as that in schemes (1) and (2).
(4) Immersion stabilization
The soil sample was immersed in treatment liquid with
the water level 1 cm higher than soil surface. Such a
treatment was carried out for five times with 2-d time
interval between two consecutive treatments.
Some treatment parameters are given in Table 1. It
should be explained here that, although treatment methods
differed, the amounts of treatment liquid applied to the sur-
face of the soil samples were similar in different schemes.
Seepage rate
The seepage rate was measured by allowing water to flow
through the sample at a constant water head and taking
readings of the amount of water flowing out of the sample,
as shown in Figure 4. The water head during the test was
maintained at 60 cm. The seepage rate was calculated as
Q sented in Figure 6. Below 2-cm depth of the samples, cal-
v¼
At
in which Q is the amount of water flowing through the soil
sample, A is the cross-section area of the sample, and t is
time duration of the test.
Strength of the surface layer
The strength of the bio-treated soil sample was measured
using a flat-bottom cylindrical penetrometer. The diameter
of the penetrometer head was 6 mm. During the measure-
ment, the penetrometer head was pushed into the sample at
a rate of 3 mm/min. At the same time, the forces and dis-
placements of the penetrometer head were recorded.
Calcite content
The calcite content of the soil sample was measured using
the EDTA titrimetric method (ISO 1984). Small pieces of
soil taken from the samples were placed in a certain volume
of hydrochloric acid. After the calcite was dissolved, the
concentration of calcium in the hydrochloric acid was meas-
ured. The amount of calcite in the small piece of soil can
then be calculated.
Volume of
treatment liquid
applied to 2cm Conversion of Times of seepage
depth (L) calcium source (%) rate reduction Surface strength (N)
2.55 34.3 8 45.13
19 2.55 49.8 8 34.69
12 2.59 89.7 379 52.29
0 2.97 58.0 70 54.24
Pore size distribution
The pore size distributions of the samples were detected
using Quantachrome Poremaster GT-60 mercury porosime-
ter. The results were presented in the form of incremental
intrusion of mercury (pore volume) against pore size.
Results
Seepage rate
Figure 5 shows the seepage rates of soil samples subject to
different levels of treatments in different treatment schemes.
In the untreated soil, the seepage rate was 0.237 L m
2 s
1.
After the treatments, the seepage rates were reduced by
8–379 times depending on the different treatment schemes.
In the schemes of surface spray and surface spray with
fibers, the seepage rate reductions were eight times. The
addition of fibers onto the soil surface did not have an evi-
dent effect on the seepage rate. In the methods of surface
spray and bulk stabilization, and immersion stabilization,
the seepage rate reductions were 379 and 70 times,
respectively.
The seepage rates against the calcium source additions in
the surface soil (2-cm range depth from the surface) are pre-
[3]
cium content was relatively low and the effect of seepage
control was weak. It can be clearly seen that the treatment
scheme (3), surface spray and bulk stabilization, was the
most effective method to reduce the seepage rate. After
three-time bulk stabilizations and one-time spray treatment,
the seepage rate was reduced to a level 25 times lower than
its original state. At this time, the amount of calcium source
used was only 0.96 mol. Further treatments also led to a sig-
nificant reduction in the seepage rate. As for the scheme of
immersion stabilization, a considerable decrease in the seep-
age rate started from the fourth treatment when the applica-
tion of the calcium source was 1.19 mol. In comparison, the
methods of surface spray and surface spray with fibers were
relatively less effective.
To explain such results, the amount and distribution of
calcite in soil and the characteristics of soil pores were
studied. The results are presented in the following
two sections.
Calcite content and distribution
The amount of calcite produced and its distribution in the
soil are the major reasons that account for the level of
the seepage rate reduction. After the bio-treatments and the
 GEOMICROBIOLOGY JOURNAL 5

(a) 1 Surface spray

0.3 Surface spray with fibers
Surface spray
Surface spray and bulk stabilization
Surface spray with fibers
Immersion stabilization

Seepage rate,v(L·m -2·s -1)

0.1
Seepage rate,v(L·m -2·s -1)

0.2

0.01

0.1
0.001

0.0001
0.0
0 4 9 14 19 0.0 0.5 1.0 1.5 2.0
Number of treatments,n(times) Calcium molar quantity n(M)
Figure 6. Relationship between calcium addition in surface soil and seepage
(b) 0.3 rate for four different treatment schemes.
Surface spray and
bulk stabilization
Seepage rate,v(L·m -2·s -1)

0.2

0.1

0.0
0 1 4 7 12
Number of treatments,n(times)

(c) 0.3
Immersion stabilization
Figure 7. The formed hard crust of soil treated by surface spray and bulk stabil-
ization: (a) the calcite layer above the original surface; (b) the cemented soil
layer below the original surface.
Seepage rate,v(L·m -2·s -1)

0.2

The distributions of calcite contents along depth in the

0.1
0.0
0 3
Number of treatments,n(times)
Figure 5. Variations of seepages rates of soils (a) surface spray and surface
spray with the addition of fibers; (b) combined surface spray and bulk stabiliza-
tion; and (c) immersion stabilization.
seepage tests, calcite contents along the depth of the samples
were measured. Figure 7 shows that, the low-permeable
hard crust at the soil surface consisted of two layers: a cal-
cite layer above the original soil surface and a cemented soil
layer below the original soil surface. The thicknesses of the
hard crusts were 10–20 mm. Below the hard crusts, soil was
not properly cemented together.
samples are given in Figure 8. In the calcite layer above the
original soil surface, calcite contents were 55–91% by weight
for different samples. In the cemented soil layer, the calcite
contents were 5–15% and decreased from the top down.
Below the hard crusts, the calcite contents were lower than
around 5%. It is also clearly seen that, the calcite content of
4 5
the sample treated by scheme (3) was greatly higher than
the other three samples. This could explain why the sample
treated by scheme (3) has the best performance in seep-
age control.
By comparing the amount of calcite to the total
amount of calcium source introduced to the soil, it can
be found that 34.3–89.7% of calcium was converted to
calcite during the treatment, as can be seen in Table 1.
For the samples treated by test schemes (1), (2), (3) and
(4), the conversion rates were 34.3%, 49.8%, 89.7% and
58.0%, respectively. Such results agreed with the observa-
tion that, obvious leakage was witnessed during the spray
treatment in schemes (1) and (2), but not in schemes (3)
and (4). This implied that the bulk or immersion
6 Y. GAO ET AL.

Figure 8. Calcite content along depth in all samples.

Table 2. Results of mercury intrusion tests.

Sample No./treatment Specific surface area Most frequent pore Pore size at 50% mercury
scheme Location Porosity (m2/g) size (lm) intrusion volume (lm)
Original sand 0.428 0.920 28.59 29.01
1 Calcite layer above sand surface 0.484 1.945 21.09 18.95
Surface soil in the hard crust 0.336 1.345 17.12 16.90
Soil below the hard crust 0.394 1.497 24.58 21.06
2 Calcite layer above sand surface 0.560 4.007 115.1 39.15
Surface soil in the hard crust 0.279 0.565 17.44 15.41
Soil below the hard crust 0.376 1.261 24.87 19.81
3 Calcite layer above sand surface 0.577 2.082 35.58 24.58
Surface soil in the hard crust 0.329 1.604 16.42 15.74
Soil below the hard crust 0.395 1.368 25.49 24.40
4 Calcite layer above sand surface 0.535 3.342 18.07 17.46
Surface soil in the hard crust 0.321 0.525 19.29 20.31
Soil below the hard crust 0.399 0.830 22.81 23.11

treatment before the application of spray treatments could
reduce the loss of the treatment liquid and increase the
treatment efficiency.
Pore size distribution
The pore size distributions of the soils were investigated
using mercury intrusion porosimetry. In the test, mercury
was pushed into a small piece of dry sample subject to step-
wise increased pressures. Under a given pressure, whether
mercury could enter a pore space was related to the size of
the pore, owing to the capillary effect. The relationship
between the pressure and the minimum size of the pores
that mercury could enter was calculated as,
4r
d¼
cos h
p
in which, d is pore diameter, r is the surface tension of
mercury, p is the pressure applied to the mercury, and h is
the contact angle of mercury on the sample. During the test,
the volume of mercury entering the sample subject to a
given pressure was recorded. In this way, the pore volume
of a specific size was obtained.
The results of the porosimetry tests are briefly presented
in Table 2. It can be seen that, both the porosities and the
pore sizes of treated samples were greatly reduced as com-
pared to the original soil. The porosity of original soil was
0.428. After the treatment, the porosities reduced to a level
0.28–0.33 in the surface soil where hard crusts were formed
and 0.37–0.39 below the hard crusts. The porosities of the
calcite layer above the soil surface were around 0.48–0.58.
Data of most frequent pore size and pore size at 50% mer-
cury intrusion volume are also given in Table 2. For all the
test samples at different locations, the pore sizes were
reduced to a level lower than original soils, except the calcite
layer in the sample with fiber addition on the soil surface.
This could be because of the interference of the fibers.
Pore size distributions obtained from the mercury intru-
[4] sion tests are often presented in the form of incremental
intrusion curves as shown in Figure 9. The vertical axis is
the volume of mercury intrusion at the specific pressure
increment, as,
Vn
Vn
1
Incremental intrusion ¼
[5]
log dn
log dn
1
in which, Vn and dn are the volume intrusion and the pore
 GEOMICROBIOLOGY JOURNAL 7

(a) (b)
1.2

Incremental intrusion,-dv/dlogd(mL·g -1)

Incremental intrusion,-dv/dlogd(mL·g -1)
1.2 Untreated sand
Untreated sand Calcite layer
Calcite layer 1.0 Cemented sand layer
1.0
Cemented sand layer Sand below the crust
Sand below the crust 0.8
0.8

0.6
0.6

0.4 0.4

0.2 0.2

0.0 0.0
1000 100 10 1 0.1 0.01 1000 100 10 1 0.1 0.01
Pore size diameter,d(µm) Pore size diameter,d(µm)
(c) (d)
1.2 1.2
Incremental intrusion,-dv/dlogd(mL·g -1)

Untreated sand

Incremental intrusion,-dv/dlogd(mL·g -1)

Untreated sand
Calcite layer
1.0 1.0 Calcite layer
Cemented sand layer
Cemented sand layer
Sand below the crust
Sand below the crust
0.8 0.8

0.6 0.6

0.4 0.4

0.2 0.2

0.0 0.0
1000 100 10 1 0.1 0.01 1000 100 10 1 0.1 0.01
Pore size diameter,d(µm) Pore size diameter,d(µm)
Figure 9. Pore size distributions curves (a) surface spray; (b) surface spray with fibers; (c) surface spray and bulk stabilization, and (d) immersion stabilization.

diameter at the applied pressure of Step n, respectively. The Penetration resistance

horizontal axis is the pore diameter in the logarithmic scale.
Penetration or bending tests were adopted to measure the
As can be seen from Figure 9, for the different samples
surface strengths of soils in previous studies (Chen et al.
and locations, the bio-treatment of soils were effective in
2015; Chu et al. 2013). In this study, a flat-end penetrometer
reducing the volume and size of pores, which was beneficial
was used to evaluate the surface strengths of the low-perme-
for the seepage control. However, at different locations of a
able hard crusts at the soil surface.
sample, the characteristics of the pore distributions were dif-
The plot of penetration force against the displacement of
ferent. In the cemented soil layer where the hard crusts were
the penetrometer is presented in Figure 10. For the original
formed, the pore size distribution curves took on a similar
soil, the penetration force was relatively small and the pene-
shape compared to the original soil. But the height of the
tration force increased to the peak value of around 6 N. For
curve became shorter and the location of the curve moved
the bio-treated samples, the forces required to penetrate the
rightwards. These results imply that the bio-treatment is
surface were much larger. The peak values were 35–55 N for
effective for different pore sizes. The soil below the hard
samples treated by different schemes. The curves show a
crust also showed the similar shape of pore distribution
drastic increase to the peak points at around 2–3 mm
curves although the level of pore size reduction was less sig-
depths, followed by a gradual decrease. However, even at a
nificant. As for the calcite layer above the soil surface, there
depth of 20 mm, the penetration forces in the treated sam-
were more narrow pores and wide pores as compared with
ples were still much larger than the original soil. It can also
the original soil. In the original soil, most of the pore sizes
be seen that the peak penetration forces treated by schemes
were in the range of 10–100 lm. In the calcite layer, the
(3) and (4) are larger than those treated by schemes (1) and
range of pore size distribution was wider.
(2). This result is consistent with the data presented in
Figures 6 and 8, which show that the samples treated by
schemes (3) and (4) have lower permeabilities and higher
calcite contents.
8 Y. GAO ET AL.

70 1.20E-04
Untreated sand
Surface spray Li, 2015
60 Surface spray with fibers 1.00E-04
Al Qabany & Soga, 2013
Surface spray and bulky stabilization

Permeability, k (m/s)
50 8.00E-05
Penetration force,F(N)

Immersion stabilization Chu et al, 2013

40 6.00E-05 Whiffin et al, 2007

30 4.00E-05 Fitting curve

y = 3 × 10 −5 x −0.743
20 2.00E-05

10 0.00E+00
0 5 10 15 20 25
0 Calcite content, w/w (%)
0 5 10 15 20
Figure 11. Relationship between the permeability and the calcite content of
Penetration depth,S(mm) uniform MICP-treated soil in previous studies (The urea concentrations in the
treatment solutions range from 0.5 to 1.5 mol/L; The mean sizes, D50, of the soil
Figure 10. Penetration force versus penetration depth curves for untreated soil
range from 0.165 to 0.420 mm).
and samples treated by four different treatment schemes.

-4
In the preliminary experiments (the data not shown in 10
the paper), we found that cracks could be easily developed
in the soil treated by the surface spray method. The cracks Calculated permeability, k (m/s)
led to the ineffectiveness of seepage control. Therefore, we
added a small amount of fibers to the soil surface before the -5
spray treatment as the so-called Treatment scheme (2) to 10
investigate whether cracks could be prevented or whether
the penetration resistance could be improved with the add-
ition of fibers. It is shown in Figure 5(a) that the samples
-6
treated by schemes (1) (without fiber) and (2) (with fiber) 10
had no significant differences in terms of the permeability
variations. Small cracks can be seen in around first 10
treatments and the cracks gradually disappear thereafter in
both samples, which is also consistent with the permeabil- -7
ity reducing trends shown in Figure 5(a). However, it 10 -7 -6 -5 -4
10 10 10 10
seems that the addition of fibers may increase the ductil-
Measured permeability, k (m/s)
ity of the soil sample. As can be seen in Figure 10, the
Figure 12. Comparison of measured and calculated permeability.
sample treated by scheme (2) (with fibers) has higher
post-peak penetration resistance that treated by scheme
(1) (without fibers). was two order-of-magnitude reduction as compared with
untreated soil. For the treated soil with non-uniform cal-
cite distribution, the permeability was calculated using the
Discussions following equation:

Permeability and calcite content H

For the MICP-treated soils, the permeability is closely
related to the amount of calcite in the soils. The relation-
ship between permeability and calcite content of uniform
MICP-treated soils was investigated in previous studies
(Al Qabany and Soga 2013; Chu et al. 2013; Li 2014;
Stabnikov et al. 2013; Whiffin et al. 2007). The results are
summarized in Figure 11. The concentrations of treatment
solution (urea) used in these studies were in a range of
0.5–1.5 mol/L. The mean sizes, D50, of the soil were in a
range of 0.165–0.420 mm. It can be seen that the perme-
ability shows a sharp decrease when the calcite content
decreased from 0 to around 2%. Beyond this range, the
variation of permeability in relation to calcite content was
moderate. The permeability could be reduced to a value
of around 1  10
6 m/s at 15% calcite content, which
k¼P   [6]
n Hi
i¼1 ki
where k and H are the overall permeability coefficient and
the depth of soil, respectively; Hi and ki are the permeability
and the soil depth of Layer i, respectively.
By using the fitting curve in Figure 11 and the equation
above, the permeability of the MICP-treated soil with the
calcite distribution shown in Figure 8 can be calculated. The
calculated and measured data are comparatively presented in
Figure 12. The calculated and measured values of permeabil-
ity are roughly consistent. The treatment effect is the best in
the treatment scheme (3), surface spray and bulk stabiliza-
tion. Such results also indicate that a calcite content larger
than 2% is favorable in terms of seepage control.

Step3: spray or immersion treatment
Hard crust
Injection well
Step1: bulk treatment
Figure 13. Schematic diagram of irrigation channels and water reservoirs construction in sandy soil ground by MICP methods.
Recommendation for future applications
For the construction of irrigation channels, a method is pro-
posed here as can be seen in Figure 13. The first step is to
apply the treatment liquid in the deep soil. The cross-well
injection method has been proved to be a feasible way for
the stabilization of deep soil (DeJong et al. 2013). This
method can also be used here. The microbial stabilization of
the soil can enhance the stability and durability of the chan-
nel during construction and services. The second step is to
excavate soil to form the designed shape of the channel. The
induced cementation effect in the first step can facilitate the
excavation because sandy soil is stabilized. The third step is
to apply treatment liquid onto the surface soil by spraying
or immersion.
It can be seen in the experimental results that the MICP
treatment of soil can effectively reduce the permeability for
the seepage control. At the same time, the penetration resist-
ance or the surface strength of soil can be improved, which
enhances the durability of the treatment effects. However,
the limitations of the method should also be addressed. The
MICP treatment usually requires at least several rounds of
treatments in order to obtain favorable permeabilities and
strengths, as can be seen in Figures 5 and 10. Currently, the
cost of materials, bacterial cultivation, as well as field appli-
cation methods are relatively high. In the MICP treatment,
ammonia is produced through the ureolytic process, which
is hazardous to the environment (DeJong et al. 2013).
Conclusions
In this study, we investigated the performance and mecha-
nisms of the MICP method for the control of water seepage
in sandy soil. The treatment was carried out by four differ-
ent application schemes, that is, (1) surface spray, (2) surface
spray with the addition of fibers, (3) surface spray and bulk
stabilization, and (4) immersion stabilization. After the
microbial treatments, tests were carried out on the bio-
treated samples to evaluate the changes in properties. These
included seepage rate tests, calcite contents and distributions
measurement, pore size distribution detection, and surface
strength tests. Through this study, the following conclusions
can be drawn,
(a) Low-permeable hard crusts can be formed at the soil
surface through MICP treatments. By applying around 2.6 L
treatment liquid onto the 2-cm deep surface soil, the seepage
rates of bio-treated samples reduced 8, 8, 379 and 70 times
GEOMICROBIOLOGY JOURNAL 9
Original sand surface
Step2: excavation Sand
Extracting well
for the samples treated by the four treatment schemes,
respectively. The conversion percentages of calcium of the
samples treated by the four different schemes were 34.3,
49.8, 89.7 and 58.0%, respectively.
(b) After the bio-treatment, the formed low-permeable
hard crusts layers were 10–20 mm in thickness and the cal-
cite content reduced from the top down. The sample treated
by scheme (3) had the thickest hard crust and highest calcite
content, which agreed with its best performance in seep-
age control.
(c) The results from the mercury intrusion tests revealed
that, in the MICP-treated soil, both pore volumes and pore
sizes decreased as compared with the original soil.
(d) In the bio-treated soil, the penetration strength and
the stiffness were much higher than those in the original
soil. The relationships between penetration forces and
depths show peak values at around 2–3 mm depths followed
by a gradual decrease in the penetration forces, which is
consistent with the calcite distributions in soils.
Disclosure statement
No potential conflict of interest was reported by the authors.
Funding
This work was financially supported by the National Natural Science
Foundation of China (No. 51608169, No. 41630638, No. 51609093, No.
51578214), the National Key Research and Development Program of
China (No. 2016YFC0800205), the Jiangsu Provincial Natural Science
Foundation of China (No. BK20150814), the 111 Project (Ministry of
Education of China, No. B13024), the Error! Hyperlink reference not
valid. (No. MOE2015-T2-2-142), and Centre for Usable Space,
Nanyang Technological University, Singapore.
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