Bacterially Stabilized Desert-Sand Bricks:

Sustainable Building Material

Vishakha Bisht 1; Leena Chaurasia 2; L. P. Singh 3; and Sanjay Gupta 4
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Abstract: This paper presents the results of an experimental investigation carried out to determine the consolidating effect of ureolytic and
nonureolytic Bacillus sp. on desert sand. Bacteria-infused desert-sand bricks were prepared using various binders (cement, lime, fly ash), and
their engineering properties were evaluated. The results showed an increase of ~19% in compressive strength and water absorption value of
17% in bacteria treated desert sand-cement bricks relative to the same properties in control. In treated bricks field emission scanning electron
microscopy (FESEM) analysis revealed thick biodepositions at points of particle to particle contact of desert sand, creating a densified
microstructure compared to untreated bricks. X-ray diffraction (XRD) and Fourier-transform infrared spectroscopy (FTIR) analysis of
bacteria-treated desert sand–cement bricks indicated the enhanced formation of other hydration products in addition to calcite, and ther-
mogravimetric (TG) analysis validated the formation of additional C─S─H (approximately 12%) and calcium hydroxide (approximately
40%). This study shows that nonureolytic bacteria–infused desert sand–cement bricks can achieve an unfired compressive strength ≥ 5 MPa
and, therefore, may serve as a sustainable alternative to other conventionally available bricks. DOI: 10.1061/(ASCE)MT.1943-
5533.0003101. © 2020 American Society of Civil Engineers.
Author keywords: Biomediated desert sand consolidation; Microbially induced calcite precipitation; Ureolytic bacteria; Nonureolytic
bacteria; Biocementation; Microstructure.

Introduction Construction biotechnology has recently emerged as an alternative

Unprecedented growth in the housing sector has brought about
a huge demand for building materials, causing a threat of the
depletion of natural resources. The demand for sustainable building
materials is a major challenge, and, over the last two decades, ef-
forts have been made to develop building materials with low em-
bodied energy processes and eco-friendly methodologies. Bricks
are the most commonly used building material, and approximately
140 billion bricks are produced annually in India (Maithel and
Uma 2000). The most common methods of brick manufacturing
involve either using a composition of cement, sand, fly ash, and
gypsum and compressing the mixture under high pressure or using
soil of appropriate physiochemical characteristics and baking up to
1,000°C. Both of these methods involve extensive energy use and,
therefore, may not be considered as sustainable building materials.
science to address issues of sustainability by harnessing natural
processes like biomineralization for consolidating sand or soil
(Stabnikov et al. 2015; Dick et al. 2006). Biomineralization is a bio-
logically induced process in which an organism creates a local micro-
environment with conditions that allow optimal extracellular chemical
precipitation of mineral phases (Ramachandran et al. 2001; Stocks
et al. 1999). The mechanism of biomineralization mediated by
ureolytic bacteria is based on urease enzyme, which catalyzes the
hydrolysis of urea into ammonia and carbon dioxide, thereby increas-
ing the alkalinity of the surrounding medium (Tittelboom et al. 2010).
The urea hydrolysis reaction and formation of calcite are as follows:
COðNH2 Þ2 þ 3H2 O → 2NHþ −
4 þ HCO3 þ OH
− ð1Þ
Ca2þ þ HCO2− −
3 þ OH → CaCO3 þ H2 O ð2Þ

Because the cell walls of the bacteria are negatively charged,

the bacteria draw cations (Ca2þ ) from the environment to deposit
1
Ph.D. Scholar, Dept. of Environmental Science and Technology, Coun- on their cell surfaces. The Ca2þ ions subsequently react with the
cil of Scientific and Industrial Research-Central Building Research Insti- CO2−3 , leading to the precipitation of CaCO3 at the cell surface that
tute, Roorkee, Uttarakhand 247667, India; Ph.D. Scholar, Uttarakhand serves as a nucleation site, according to the following equations:
Technical Univ., Dehradun, Uttarakhand 248001, India (corresponding
author). Email: vishakha.bisht27@gmail.com
2
Ca2þ þ Cell → Cell-Ca2þ ð3Þ
Scientist, Dept. of Environmental Science and Technology, Council of
Scientific and Industrial Research-Central Building Research Institute,
Cell-Ca2þ þ CO2−
3 → Cell-CaCO3 ↓ ð4Þ
Roorkee, Uttarakhand 247667, India. ORCID: https://orcid.org/0000-0001
-8204-918X. Email: leenachaurasia@yahoo.co.in
3
Scientist, Dept. of Environmental Science and Technology, Council of
Nonureolytic bacteria, however, form calcium carbonate by di-
Scientific and Industrial Research-Central Building Research Institute, rect and indirect means (Jonkers 2011; Jonkers et al. 2010). Direct
Roorkee, Uttarakhand 247667, India. Email: lpsingh@cbri.res.in CaCO3 precipitation by bacteria is due to the metabolic conversion
4
Professor, Dept. of Biosciences, Swami Rama Himalayan Univ., of calcium lactate, as per the following equation:
Dehradun, Uttarakhand 248016, India. Email: sanjaygupta@srhu.edu.in
Note. This manuscript was submitted on June 5, 2018; approved on CaC6 H10 O6 þ 6O2 → CaCO3 þ 5CO2 þ 5H2 O ð5Þ
September 5, 2019; published online on March 25, 2020. Discussion period
open until August 25, 2020; separate discussions must be submitted for Indirect formation is due to the reaction of metabolically pro-
individual papers. This paper is part of the Journal of Materials in Civil duced CO2 molecules with CaðOHÞ2 (portlandite) present in the
Engineering, © ASCE, ISSN 0899-1561. concrete matrix, according to the following equation:

© ASCE 04020131-1 J. Mater. Civ. Eng.

J. Mater. Civ. Eng., 2020, 32(6): 04020131

 CO2 þ CaðOHÞ2 → CaCO3 þ H2 O ð6Þ Table 1. Physicochemical properties of desert sand
Physical properties and composition (mass %) Results obtained
Current studies have also found the potential for bacterial cells Bulk density (g=cm3 ) 2.1
to sequester atmospheric CO2 and fix it into different carbonate Specific gravity 2.7
minerals such as calcite, magnesite, and dolomite (Bharti et al. Water absorption (%) 0.4
2014; Dhami et al. 2014). Moisture content (%) 0.1
Over the last few years, the potential of biomineralization has pH 9.5
been widely recognized in petroleum, geological, and civil engi- Sulphate content (%) 2.9
neering (Dejong et al. 2006; Ivanov and Chu 2008). Recently, a Chloride content (%) 0.01
Total dissolved solvent —
number of studies have confirmed that microbially induced calcite
Organic impurities —
precipitation (MICP) has the potential to alter soil characteristics, Clay + silt (%) 9.3
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leading to increased shear strength and stiffness while maintaining Clay (%) 8.5
permeability (Warren et al. 2001). This method is useful for tun- Silt (%) 0.8
neling, earthquake repair, and instant pavements (Van Paassen Sand (%) 90.7
2009; Dejong et al. 2010). Researchers have also worked on bio-
aggregation of soil particles and biocrusting of the soil surface in
order to control water and wind erosion of soil (Bang et al. 2011;
Stabnikov et al. 2011). The performance of a ureolytic Bacillus spe- Table 2. Sieve analysis of desert sand
cies has been evaluated for stabilization of desert sand, and high IS sieve Retained
resistance to wind erosion by a 33-ms−1 wind speed was observed 4.75 mm —
(Chen et al. 2016). Scale model tests have demonstrated the effec- 2.36 mm —
tiveness of MICP in improving resistance in sandy soil (Martinez 1.18 mm 0.45% (4.5 g)
et al. 2013) and resistance to silty sand liquefaction (Mortensen 600 μm 1.3% (13 g)
and DeJong 2011). Biocementation techniques have also been 300 μm 3.0% (30 g)
suggested for slope stabilization, strengthening coastlines from 150 μm 94.4% (944.0 g)
progressive erosion (Emmanuel et al. 2016; Montoya et al. 2013).
Other applications of MICP include bioremediation of polluted
soil in order to immobilize pollutants in the soil before construc- Table 3. Physicochemical properties of ordinary portland cement and fly
tion (Warren et al. 2001; Fujita et al. 2000) and production of ash
bricks [Sarda et al. 2009; Dhami et al. 2012; Bernardi et al.
Composition (mass %) and physical properties Cement Fly ash
2014; (G. K. Dosier, “Methods for making construction material
using enzyme producing bacteria,” US Patent No. 8,728,365 SiO2 19.3 56.8
(2014)]. Al2 O3 5.8 30.2
Previous work done on masonry sand consolidation with lime Fe2 O3 5 4.9
(10%–27% by weight) reported a strength of 1.0 MPa, while with CaO 64.3 0.8
MgO 0.8 0.7
cement (12%–19% by weight) strength was found to increase to
Alkalis (K2 O þ Na2 O) 0.9 1.4
2.5 MPa. However, the infusion of B. pasteurii in masonry sand SO3 — 0.2
resulted in a strength of 1–2 MPa (Bernardi 2012). Work on Others 1.5 4.9
B. megaterium–infused soil-cement bricks reported a compressive Loss on ignition 4 3.6
strength of approximately 6–7 Mpa; the strength of control bricks Specific gravity 3.13 2.2
was 6.0 MPa. In addition, a significant reduction in water absorp- Consistency (%) 27 —
tion was observed (Mukherjee et al. 2013). In both studies, ureo- Initial setting time (min) 170 —
lytic bacteria was used, and regular treatment/spraying of bricks Final setting time (min) 240 —
with cementation media was conducted for a period of 28 days. Specific surface area (m2 =g) 315.8 252
In the present study, an effort has been made to develop building Particles retained on 45 μm (%) — 42
bricks using desert sand and biocementation techniques in order to
prepare sustainable bricks. Moreover, in this study, nonureolytic
bacteria (B. cohnii) was incorporated, and a comparative study with Cement, Fly Ash, and Lime
ureolytic bacteria (B. megaterium) was carried out in order to de-
termine which bacterial species shows better cementation potential Ordinary portland cement (OPC) with a Blaine’s fineness of
in desert sand. 390 m2 =kg, low-calcium class-F fly ash procured from the national
thermal power plant (NTPC) in Dadri (Uttar Pradesh, India), and
class-B hydrated lime (70% calcium hydroxide) were used for the
Materials and Methods present study. The physicochemical properties of cement and fly
ash are presented in Table 3; these properties were determined
in accordance with IS 8112 (IS 1989) and IS 3812 Part 2 (IS
Desert Sand 2013), respectively. The chemical composition of hydrated lime
The desert sand used in the present study was obtained from is shown in Table 4; this was determined in accordance with the
the Rajasthan area in India and is abundantly available. The requirements of IS 712 (IS 1984).
physicochemical properties of desert sand are presented in Table 1,
and gradation from sieve analysis is shown in Table 2; these prop-
Bacteria and Growth Conditions
erties were determined in accordance with the requirements of
Indian standard (IS) 2386 (IS 1963a, b) and IS 383 (IS 1970), For the present study, calcite-precipitating ureolytic bacteria,
respectively. B. megaterium, and nonureolytic bacteria, B. cohnii, were procured

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 Table 4. Chemical characterization of hydrated lime briquettes were cast using normal tap water. The mix was placed
Composition Lime (%) on the mold base; river sand was sprinkled on the top layer in order
to prevent the mix from sticking to the mold compressor, onto
CaO 70
which a load of 10 tons was applied. The briquettes were then cured
SiO2 1.9
Al2 O3 0.6
in moist conditioning by wet jute bag for a period of 28 days. After
MgO 1.5 completion of the curing period, the briquettes were tested on a
Fe2 O3 0.5 universal testing machine. In addition, bacteria-treated briquettes
SO3 0.2 were cast at two different applied loads, 10 and 15 tons. Compres-
Alkalis 5.1 sion testing of these briquettes was done in order to optimize the
Others 2.5 applied load at which further casting would be performed. Like-
LOI 17.7 wise, desert sand–lime bricks were prepared by combining desert
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Note: LOI = loss on ignition.
from microbial type culture collection (MTCC) (MTCC 10086 and
10221, respectively), Chandigarh, India. The nonureolytic and ure-
olytic bacteria were cultured in a nutrient broth and a urea-based
broth, respectively. The cultures were incubated aerobically in a
shaker incubator at 30°C and 37°C at 120 rpm for 24 and 48 h,
respectively. The bacteria were incubated until the cultures ob-
tained an optical density of approximately 1.5 or a cell concentra-
tion of 107 cells=mL at a 600 nm wavelength.
Preparation of Briquette and Brick Specimens
A preliminary study was done on briquettes with dimensions of
100 × 50 × 50 mm; after observing the results (compressive
strength, water absorption) of ureolytic and nonureolytic bacteria–
treated desert-sand briquettes, bricks of standard size (230×
110 × 76 mm) were cast in order to evaluate their engineering
properties and to analyze microstructural modifications after bac-
terial infusion. Four different mixes were prepared using raw
materials—namely, cement, lime, and fly ash—as binders to the
desert sand. Different series for five mixes were prepared to have
varying percentages of raw materials, and six briquette specimens
(untreated and bacteria-treated) per series were cast.
Table 5 presents various series of the five mix compositions; the
amount of raw materials required for the casting of six briquettes is
shown. For the casting of six briquettes, the dry mix weighed
3,000 g, and 168 mL of water/bacterial culture was required for
preparing untreated/treated specimens. In the first mix, cement
was added to desert sand in quantities of 5%, 8%, 10%, 12%,
and 15% by weight and mixed manually in order to prepare
five different variations of desert sand–cement mix. Ureolytic/
nonureolytic bacterial culture was added to the desert sand–cement
dry mix to cast bacterial briquettes (100 × 50 × 50 mm); control
sand with varying amounts of hydrated lime in order to prepare
three different series of the mix. Lime was added to desert sand
in quantities of 15%, 20%, and 25% by weight, and mix was pre-
pared using a nonureolytic bacterial culture with 107 cells=mL. A
third mix of desert sand–lime–cement briquettes was prepared by
combining desert sand with 5% cement (by weight); lime was
added in quantities of 8%, 10%, 12%, and 15% by weight in order
to prepare four different series of the mix; and then nonureolytic
bacteria was added. The fourth mix used 15% fly ash and 5% ce-
ment (by weight), and bacterial culture was added to 80% desert
sand. The fifth and final mix was similar to the fourth mix, differing
only in that cement was replaced by lime (5%).
Nonureolytic bacteria–treated and untreated desert sand–cement
bricks of standard size (230 × 110 × 76 mm) were prepared after
evaluating the engineering properties of the briquettes (100×
50 × 50 mm). Cement was added to desert sand in quantities of
5%, 8%, 10%, 12%, and 15% by weight, and bacterial culture/
water was incorporated into the desert sand–cement dry mix in or-
der to make a homogenous mixture. Bricks were cured in moist
conditioning at a temperature of 25°C  3°C by wet jute bag
for 28 days, and compressive strength tests were carried using a
universal testing machine (UTM) (UH-1000knI, Shimadzu, Kyoto,
Japan) with a capacity of 1,000 kN.
Measurement Methods
Compressive Strength
Compression testing was performed on all the briquettes/bricks in
accordance with IS 3495 Part 1 (IS 1992) after 28 days of curing by
water spray. Extended curing of a few bricks was done for 28 days
by the water ponding method, and the compression test was re-
peated in order to evaluate the strength (Rai et al. 2013). The frogs
were filled with desert sand and thin plywood sheets were placed
on both faces of the bricks. The test was performed until failure of a

Table 5. Mix design of various series of five desert sand mixes with cement, lime, fly ash and bacteria for treated briquettes or water for control briquettes
Brick mix Composition/series (%) Desert sand (g) Cement (g) Bacteria/water (ml) Lime (g) Fly ash (g)
Desert sand–cement 5 2,870 150 168 — —
8 2,760 240 168 — —
10 2,700 300 168 — —
12 2,640 360 168 — —
15 2,550 450 168 — —
Desert sand–lime 15 2,550 — 168 450 —
20 2,400 — 168 600 —
25 2,250 — 168 750 —
Desert sand–lime–cement 8 2,610 150 168 240 —
10 2,550 150 168 300 —
12 2,490 150 168 360 —
15 2,400 150 168 450 —
Desert sand–fly ash–lime 15 2,400 — 168 150 450
Desert sand–fly ash–cement 15 2,400 150 168 — 450

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 brick specimen, and the ultimate crushing load was recorded in or-
der to calculate the strength of the bricks.
Water Absorption (%) in Bricks
The control and bacteria-treated desert sand–cement bricks and
desert sand–lime–cement bricks were tested for water absorption
after 28 days of casting/water spray as per IS 3495 Part 2 (IS 1992).
Characteristic Techniques
Bacteria-treated and untreated desert sand–cement briquette spec-
imens were analyzed through various techniques in order to inves-
tigate biomediated mineral formation and consequent filler effects
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of the bacteria. The morphological features and mineralogical com-
position of desert sand-bacterial and control bricks were analyzed
using field emission scanning electron microscopy (FESEM) (TES-
CAN MIRA 3) and energy-dispersive X-ray spectroscopy (EDX)
(X-flash detector 5010, Bruker, Nano GmbH, Berlin), respectively.
The brick samples were dried at room temperature, crushed, and
gold coated with a JFC-1200 fine coater prior to examination.
X-ray diffraction (XRD) spectra were obtained using a Rigaku
DMax 2200 diffractometer with a Cu anode at 40 kV=40 mA,
scanning from 3° to 80° 2θ with a scanning rate of 0.5°=min
throughout the experiment. Fourier-transform infrared spectros-
copy (FTIR) (ThermoNicolet NEXUS) was used for the study of
chemical bonding within the bacteria-infused desert sand–cement
matrix. The measurement range was between 400 and 4,000 cm−1 ,
and all brick specimens were scanned 64 times with a resolution of
2.0 cm−1 . Thermogravimetric (TG) analysis was carried out to
quantify the compounds formed in the biobricks as a result of bio-
cementation activity. TG (Perkin-Elmer Diamond) studies were
performed at a heating rate of 5°C=min under nitrogen flow. Quan-
tification of CH (calcium hydroxide), CC (calcium carbonate) was
calculated directly from the TG curves using the following equa-
tions (Lam et al. 2000; Aono et al. 2007):
CHð%Þ ¼ WLðCHÞð%Þ × MWðCHÞ=MWðHÞ ð7Þ
where WL(CH) = weight loss attributable to CH dehydration; and
MW(CH) and MW(H) = molecular weights of CH (74 g=mol) and
water (18 g=mol), respectively.
CCð%Þ ¼ WLðCCÞð%Þ × MWðCCÞ=MWðCO2 Þ ð8Þ
where WL(CC) = weight loss attributable to CC decarboxylation;
and MW(CC) and MWðCO2 Þ = molecular weights of CC (100 g=
mol) and CO2 (44 g=mol), respectively. After 28 days, C-S-H
formed due to bacterial activity in the bricks; varying amounts
of cement were quantified using the following equation by TG
analysis (Singh et al. 2015):
C─S─Hð%Þ ¼ Total LOI − LOIðCHÞ − LOIðCCÞ ð9Þ
where LOI(CH) = dehydration of CH around 400°C–500°C;
and LOI(CC) = decarbonation of calcium carbonate around
600°C–800°C.
Results and Discussion
Measurement Methods
Compressive Strength
The 28-day compressive strength of nonureolytic bacteria–infused
desert sand–cement briquettes showed an enhancement of approx-
imately 35%, 32%, 20%, 19%, and 6% compared to control for
samples with 5%, 8%, 10%, 12%, and 15% cement, respectively
© ASCE 04020131-4
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Fig. 1. Compressive strength of bacteria-treated (ureolytic/
nonureolytic) and untreated/control desert-sand bricks (100 × 50 ×
50 mm) with varied percentages of cement after 28 days of casting.
(Fig. 1). In comparison to control (untreated specimens), the non-
ureolytic bacteria treated specimens showed a consistent increase in
strength, with an increase in the percentage of cement (5%–15%).
Ureolytic bacteria–infused bricks showed strength equivalent to
control for specimens with 5% and 8% cement; however, enhance-
ments in strength were observed in specimens with cement ≥ 10%.
Ureolytic bacteria–infused briquettes with 10%, 12%, and 15% ce-
ment showed enhancements in strength of approximately 9%, 7%,
and 26%, respectively, compared to control. Thus, the ureolytic
bacteria requires approximately 10% cement (optimum) for medi-
ating biomineralization within the sand–cement matrix. Moreover,
out of the two Bacillus species, the nonureolytic bacteria showed
better biocementation activity and, therefore, higher strength than
the ureolytic bacteria in the desert sand–cement briquettes. Overall,
the results validated that bacteria increase the packing density of
desert sand–cement briquettes by clogging the pores (filler effect)
with biomediated mineral depositions, leading to the enhancement
of the compressive strength. After 28 days of water immersion (water
ponding method), further enhancement in compressive strength
was noticed in both control and bacterial briquettes as compared
to wet compressive strength after 28 days of water spray. Si-rich
halos are more resistant to carbonatation than cement paste; in con-
tact with dolerite (feldspar, pyroxene, olivine, quartz), the paste
provides calcium-alumina-silicate-hydrate (CASH) and C4 AHn ·
C4 AHn is known to be metastable with respect to hydrogarnet
and, in fact, converts slowly to hydrogarnet during cement curing
(Lea 1970). Hence, the reason for the strength enhancement in the
desert sand–cement system after extended curing of 28 days can be
attributed to the formation of CASH and C─S─H (Jain 2016). The
results revealed that even after extended curing, bacterial briquettes
gained more strength than control briquettes (Fig. 2), indicating the
accelerated formation of hydration products in bacteria-treated des-
ert sand–cement briquettes. The nonureolytic bacteria–treated bri-
quettes cast at 10 tons of applied load showed an approximately
10% enhanced compressive strength compared to briquettes cast
at 15 tons of applied load (Fig. 3). At high applied load, the effec-
tive cohesion between the desert sand particles due to decreased
interparticle spaces seems to have had a negative impact on the
strength of the bricks. The primary restriction on microbial trans-
port is the size of pore throats within the soil matrix; the microbes
must pass through the pore throats as they move from one pore
space to another (Mitchell and Santamarina 2005). The decrease
in pore throat space (unoccupied) is believed to hinder bacterial
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Fig. 2. Compressive strength of bacteria-treated (ureolytic/
nonureolytic) and untreated/control desert-sand bricks (100 × 50 ×
50 mm) with varied percentages of cement after 28 days of water
immersion.
percolation inside the matrix. Sieve analysis (Table 2) shows that
the desert sand belongs to grading zone IV, with particle sizes in the
range 150–300 μm. With the application of a high load (15 tons),
the interparticle spaces are enormously decreased (pore throats),
which consequently decreases the fraction of bacteria at particle–
particle contacts and minimizes bacterial activity. However, in bri-
quettes cast at a lower applied load, effective biomineralization at
particle–particle contacts occurs due to the unhindered motion of
the bacteria. Therefore, such bricks attain a higher compressive
strength due to the presence of a large fraction of bacteria and con-
sequent heightened mineralization, which increases the packing
density of the bricks.
Taking into consideration the greater potential of nonureolytic
bacteria over ureolytic bacteria, the remainder of the experimenta-
tion was carried out by incorporating the former in desert sand.
Fig. 4 shows that bacterial infusion in lime–desert sand bricks in-
creased their compressive strength by up to approximately 38%;
however, the mix did not yield enough strength. In other words,
the absolute strength of this specific mix was low, between 0.5
and 1 MPa. In bacteria-infused lime–cement–desert sand bri-
quettes in which the amount of cement was kept constant (5%)
and the amount of lime was varied (8%–15%), an enhancement
Fig. 3. Compressive strength of bacteria-treated and untreated desert-
sand bricks (100 × 50 × 50 mm) with varied percentages of cement at
10 and 15 tons of applied load.
© ASCE 04020131-5
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Fig. 4. Compressive strength of nonureolytic bacteria–treated and con-
trol bricks (100 × 50 × 50 mm) with varied percentages of lime.
in compressive strength of approximately 55% was observed in
comparison to untreated specimens (Fig. 5). From these results,
it can be deduced that a lime–cement combination in bacteria-
infused desert-sand bricks may be used in order to reduce cement
consumption and the relative cost, because only 5% cement
was used.
Bacteria-infused fly ash–lime–desert sand briquettes showed an
18% enhancement in strength relative to control, and bacteria-
infused fly ash–cement–desert sand briquettes showed a 23.6%
higher strength relative to its respective control (Fig. 6). Further-
more, bacteria-infused fly ash–cement–desert sand briquettes also
exhibited heightened strength compared to bacteria-infused fly
ash–lime–desert sand briquettes.
The standard size bricks prepared using nonureolytic bacteria–
treated desert sand–cement showed an increase of approximately
23%, 21.9%, 20.4%, 16.6%, and 17.3% in compressive strength
compared to control bricks with 5%, 8%, 10%, 12%, and 15% ce-
ment, respectively (Fig. 7). The increase in strength can be attrib-
uted to bacteria-facilitated precipitation (biodeposition), which
densifies the desert sand–cement matrix and results in an enhance-
ment in overall strength compared to control bricks. Moreover,
the results reveal the enhanced biocementation potential of nonur-
eolytic bacteria relative to ureolytic bacteria and also show that
Fig. 5. Compressive strength of nonureolytic bacteria–treated and con-
trol bricks (100 × 50 × 50 mm) with varied percentages of lime and
5% cement.
J. Mater. Civ. Eng.

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Fig. 6. Compressive strength of bacteria-treated and control desert-
sand bricks (100 × 50 × 50 mm) with 5% lime/cement and 15%
fly ash.
Fig. 7. Compressive strength of nonureolytic bacteria–treated and un-
treated desert-sand bricks (230 × 110 × 76 mm) with varied percen-
tages of cement after 28 days of casting.
calcifying bacteria can perform biomineralization even in the ab-
sence of chemical feed.
Water Absorption
Nonureolytic bacteria-infused desert sand–cement bricks had water
absorption values of 17%, while untreated brick specimens had
water absorption values of approximately 19%. Similarly, untreated
desert sand–cement briquettes had water absorption values of ap-
proximately 14%, while ureolytic and nonureolytic bacteria–treated
briquettes had water absorption values of approximately 12% and
11%, respectively (Table 6). This indicates that bacteria-induced
mineral deposition filled the pores inside the desert sand–cement
matrix and acted as a barrier to the penetration of water, resulting
in low water absorption values. Table 7 shows the water absorption
of nonureolytic bacteria–infused desert sand–lime–cement (5%)
briquettes; a substantial reduction in water absorption (approxi-
mately 8%) was observed. The reduction in water absorption clearly
indicates that lime can be utilized in order to reduce the amount of
cement in bacteria-infused desert-sand bricks.
Characteristic Techniques
FESEM/EDX Analysis
FESEM micrographs of ureolytic/nonureolytic bacteria–treated/
untreated desert sand–cement brick specimens reveal a distinct
© ASCE
J. Mater. Civ. Eng., 2020, 32(6): 04020131
Table 6. Water absorption (%) in untreated and bacteria-treated desert
sand–cement bricks (100 × 50 × 50 mm)
8% 10% 12% 15%
Brick specimens cement cement cement cement
Control 15.6 15.5 14.0 12.3
Ureolytic bacteria briquette 13.8 12.6 12.2 11.2
Nonureolytic bacteria briquette 13.8 13.2 9.9 8.5
Table 7. Water absorption (%) in untreated and bacteria-treated desert
sand–lime–cement bricks (100 × 50 × 50 mm)
Brick 8% lime + 10% lime + 12% lime + 15% lime +
specimens 5% cement 5% cement 5% cement 5% cement
Control 15.3 13.6 12.7 13.8
Nonureolytic 14.7 12.6 11.8 12.1
bacteria briquette
gap between sand particles in the untreated brick sample [Fig. 8(a)].
However, bacteria-treated brick micrographs [Figs. 8(b and c)]
show thick biodepositions that bind the sand particles and fill the
gaps between them, which results in densification of the sand–
cement matrix. The bacterially cemented area containing precipi-
tated minerals between the desert sand particles was focused, and
multipoint EDX of treated bricks was performed. The results
showed Ca=Si ¼ 1.5 and 0.9 in ureolytic bacteria–treated and non-
ureolytic bacteria–treated brick, respectively, indicating increased
formation of hydrated products within the desert sand–cement sys-
tem. The products that formed as a result of bacterial mineralization
filled the voids, increased the packing density within the cement–
sand matrix, and thereby improved the overall engineering proper-
ties of the bricks.
XRD Analysis
XRD analysis validated the biocementation process and depicted
the additional formation of C─S─H and calcite in bacteria-infused
desert sand–cement bricks (Fig. 9). The peaks at 2θ ¼ 20.82° and
2θ ¼ 26.6° represent quartz, which is the major component of des-
ert sand. Calcite peaks at 2θ ¼ 29.58° and 39.53° and aragonite
peaks at 2θ ¼ 36.53° and 45.83° were found in both control and
bacterial samples; however, the intensity of the peaks in bacterial
specimens was higher than in untreated specimens, indicating the
formation of calcium carbonate by the bacteria.
Dissolution of minerals such as quartz, feldspar, mica, and do-
lomite plays an important buffer role with respect to the pH, and
their dissolution liberates elements that favor the precipitation of
certain phases (zeolites, CSH, CASH) (Eric and Philippe 2006).
Therefore, in the desert sand–cement system, phases like zeolite
and C─S─H were observed. In addition to calcite, afwillite (a form
of C─S─H) peaks corresponding to 2θ ¼ 27.96° were found with
extremely high intensity in bacterial specimens (both ureolytic/
nonureolytic) relative to control specimens. Moreover, a few addi-
tional peaks of C─S─H at 2θ ¼ 27.08°, 2θ ¼ 31.5°, and 2θ ¼ 33°
and gyrolite (a form of C─S─H) at 2θ ¼ 42.45° were observed in
bacterial specimens with comparatively high intensity relative to
control specimens, confirming the additional formation of hydra-
tion products in bacterial bricks. From these results, it may be de-
duced that, aside from calcite, bacteria accelerate the additional
formation of hydration products by serving as nucleating sites dur-
ing the mineralization process. The XRD spectra also revealed that
if the intensity of calcite is weak in any specific composition (com-
prising of 8%, 10%, 12%, or 15% cement), the intensity of C─S─H
was stronger and vice versa.
04020131-6 J. Mater. Civ. Eng.
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Fig. 8. FESEM showing microstructure of (a) untreated; (b) ureolytic bacteria–treated; and (c) nonureolytic bacteria–treated desert sand–cement
system. EDX analysis is included for bacteria-treated specimens.

FTIR Analysis peak at approximately 870 cm−1 and around 1,400–1,430 cm−1
The FTIR spectroscopy (Fig. 10) highlighted the presence of represents CO2 vibration of calcium carbonate (Horgnies et al.
quartz, which was detected by its characteristic peak at approxi- 2013; Carrasco et al. 2012). In bacteria-treated specimens (B2,
mately 778 cm−1 in desert sand (Varlikli et al. 2009). However, the B3), an increase in the intensity of the peak at approximately

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Fig. 9. X-ray diffraction analysis of untreated, ureolytic bacteria–treated, and nonureolytic bacteria–treated desert sand–cement bricks with
(a) 8% cement; (b) 10% cement; (c) 12% cement; and (d) 15% cement.

870 cm−1 and a broadening of the peak around 1,430 cm−1 was
observed, indicating increased production of calcite by biomin-
eralization. C─S─H, which is characterized by the major band
at 1,030–970 cm−1 and around 1,030 cm−1 (Si─O asymmetric
stretching vibration), is distinctly visible in B1 (untreated desert
sand–cement brick). In the bacterial bricks, a shift of bands to
higher wave numbers was observed in bacterial specimens due to
the polymerization of silica. The shift in the bands in this region
indicates polymerization of silica; the higher the wave number, the
more polymerization (Carrasco et al. 2012). In this study, the reason
for an increase in wave number and the formation of additional
C─S─H may be bacteria-mediated nucleation, which facilitates
the polymerization of silicate chains on calcium-adhered bacterial
cell walls. Water absorption bands were observed in the region
of 1,593–1,645 cm−1 in B1 due to H─O─H bending vibrations of
H2 O molecules. In B2 (nonureolytic bacteria–infused brick) and
B3 (ureolytic bacteria–infused brick), the peak is distorted, indicat-
ing the enhanced formation of hydrated products in which water is
consumed. It can be concluded that in bacteria-treated bricks, the
growth of hydrated products along with increased calcite precipi-
tation results in densification of the microstructure and improve-
ment in macroproperties.
TG Analysis
The TG curves (Fig. 11) show the weight loss (%) of untreated and
bacteria-treated desert sand–cement bricks (having varying cement
percentages) at different temperatures. The TG analysis was carried
out in order to quantify the compounds formed within the bacteria-
treated desert sand–cement matrix due to bacterial activity. The re-
sults showed a maximum amount (% by weight) of CC formed
within desert sand–cement bricks, followed by C─S─H and CH
(Fig. 12). The percent by weight of CC in bacterial samples
was found to be relatively higher than in control samples due to
MICP in bacteria-treated bricks, that is, approximately 5% higher
than control in case of nonureolytic bacteria–treated brick and ap-
proximately 8% higher than control in ureolytic bacteria–treated
brick. Likewise, bacteria-treated desert sand–cement bricks showed
a heightened amount of C─S─H, that is, approximately 15%
(nonureolytic bacteria–treated bricks) and approximately 10%
(ureolytic bacteria–treated bricks) more percent by weight than un-
treated specimens, indicating the additional formation of C─S─H.
Moreover, as compared to control, the nonureolytic bacteria–
treated and ureolytic bacteria–treated bricks showed approximately
60% and 30% more CH (% by weight), respectively, in desert-sand
brick.

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J. Mater. Civ. Eng., 2020, 32(6): 04020131

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Fig. 10. FTIR spectra of B1 (untreated), B2 (nonureolytic bacteria–treated), and B3 (ureolytic bacteria–treated) desert sand–cement bricks.

Fig. 11. TG curves showing weight loss (%) in untreated, ureolytic bacteria–treated, and nonureolytic bacteria–treated desert sand–cement bricks
having different percentages of cement.

The percent by weight of CC was not high in 8% and 10%
bacteria-infused desert sand–cement bricks; however, in such
bricks other hydration products such as CH and C─S─H were de-
tected in relatively high amounts. This indicates that in bacteria-
treated brick specimens, aside from the formation of calcite through
MICP, bacteria also facilitate precipitation of other mineral phases;
therefore, higher amounts of other hydration products were also
observed. Furthermore, bacteria mineralize calcite or other hydra-
tion products in different amounts depending on the availability of
different negative ions (CO2− − 2−
3 , OH , and H2 SiO4 ) in the vicinity
of the bacterial cells.
Due to its numerous consequences, MICP, so-called, carbonato-
genesis has attracted much attention from both basic and applied
points of view (Rodriguez et al. 2003). In previous studies, MICP
has been considered as a sole technique by which bacteria precipi-
tate calcite on cell walls and fill pore spaces within the cement–
sand matrix. Based on the results of the analysis of macroproperties
and microstructure and the quantitative analysis of bacteria-infused
desert sand–cement bricks in the present study, it is found that
heightened amounts of not only calcite but also of other minerals—
namely, calcium silicate hydrate and calcium hydroxide—are
formed. Calcifying bacteria provide a large surface area and serve
as heterogeneous nucleation sites, which facilitates the adhesion of
numerous Ca2þ ions and the consequent formation of calcite and
other mineral phases (hydrated products) on cell walls. The bacteria
actively proliferate within the sand–cement matrix and deposit the

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Downloaded from ascelibrary.org by University of Massachusetts, Amherst on 03/27/20. Copyright ASCE. For personal use only; all rights reserved.
Fig. 12. Amount of (a) CC; (b) C─S─H; and (c) CH in untreated, ureolytic bacteria–treated, and nonureolytic bacteria–treated desert sand–cement
bricks having different percentages of cement.
minerals formed over their cell walls. Bacterially formed mineral
deposits, often called biocement, fill up the pore spaces within the
desert sand–cement/lime matrix; in other words, it acts as a binder
between loose substrate particles and leads to the stabilization of 2.
the desert sand.
Conclusions
In the present work, an effort was made to harness a natural bio-
logical process for consolidating abundantly available desert sand 3.
for use in bricks by using a low percentage of carbon-intensive
materials (i.e. lime, cement, fly ash) to replace soil–cement bricks
and other conventionally available bricks. Fired clay bricks carry
around 3 MJ=kg of embodied energy; in this study, unfired bricks
were prepared. This is expected to reduce the embodied energy
and, consequently, the cost of the bricks. Moreover, bacteria-treated
desert sand–cement bricks may serve as a viable alternative to other 4.
unfired bricks (fly ash–lime and fly ash–cement bricks, etc.) be-
cause of improved engineering properties and relatively cheap con-
stituents. Nonureolytic bacteria are known to harness atmospheric
CO2 for calcite precipitation by facilitating its reaction with por- 5.
tlandite; therefore, a decreased carbon footprint is also expected.
Stabilization of desert sand using microbes may prove to be a sus-
tainable approach and expected to make an appropriate utilization
of desert sand in the form of building bricks.
The following summarizes the primary observations made in
this study: 6.
1. Compared to untreated standard-sized bricks with 5%, 8%,
10%, 12%, and 15% cement, nonureolytic bacteria–desert
sand–cement bricks showed an increase of approximately
© ASCE 04020131-10
J. Mater. Civ. Eng., 2020, 32(6): 04020131
23%, 22%, 20%, 16.6%, and 17%, respectively, in compres-
sive strength and a low water absorption value of approxi-
mately 17%.
Between the two chosen bacterial species, the nonureolytic bac-
teria (B. cohnii) exhibited better cementation potential and
mineralized even in absence of chemical feed (i.e., calcium lac-
tate). Therefore, nonureolytic bacteria may be preferred over
ureolytic bacteria (B. megaterium) for the production of desert
sand–based bricks.
The results imply that the utilization of nonureolytic bacteria in
preparing desert-sand bricks can eliminate the need for urea, cal-
cium chloride incorporation in growth medium, or periodic ce-
mentation treatment (as in case of ureolytic bacteria). Moreover,
there will likely be no release of ammonia to the environment;
the present methodology also eradicates regular infusion of bac-
terial culture into the bricks after casting.
Microscopy techniques reveal that biologically driven deposi-
tion occurs preferentially near particle–particle contacts within
the desert sand–cement matrix. This can be considered a major
reason for the binding of loose particles of desert sand.
FTIR and XRD analysis revealed the presence of additional
C-S-H formed along with calcite due to bacteria-mediated
mineralization. Likewise, TG analysis of bacteria-treated bricks
showed higher amounts of hydrated products as well as calcite
compared to untreated bricks, validating the results of XRD and
FTIR analysis.
The nucleation and cementation effects of calcifying bacteria
result in the formation of mineral phases that bind loose desert
sand particles; this leads to the densification of the desert sand–
cement/lime matrix.
J. Mater. Civ. Eng.
 Acknowledgments
The authors gratefully acknowledge the financial support of the
Uttarakhand Council of Biotechnology, Uttarakhand, India. Any
opinions, findings, and conclusions or recommendations expressed
in this material are those of the writer(s) and do not necessarily
reflect the views of the Uttarakhand Council of Biotechnology.
Supplemental Data
Figs. S1 and S2 are available online in the ASCE Library (www
Downloaded from ascelibrary.org by University of Massachusetts, Amherst on 03/27/20. Copyright ASCE. For personal use only; all rights reserved.
.ascelibrary.org).
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