SPE-191778-MS

Self-Healing Biocement and Its Potential Applications in Cementing and

Sand-Consolidation Jobs: A Review Targeted at the Oil and Gas Industry

C. I. Noshi and J. J. Schubert, Texas A&M University

Copyright 2018, Society of Petroleum Engineers

This paper was prepared for presentation at the SPE Liquids-Rich Basins Conference-North America held in Midland, TX, USA, 05-06 September 2018.

This paper was selected for presentation by an SPE program committee following review of information contained in an abstract submitted by the author(s). Contents
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consent of the Society of Petroleum Engineers is prohibited. Permission to reproduce in print is restricted to an abstract of not more than 300 words; illustrations may
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Abstract
There are several self-healing mechanisms, both natural and artificial, applied to cementitious materials.
In recent years, microbially induced calcite precipitation (MICP) technology has garnered special attention
in the fields of Microbiology and Civil Engineering. The technology involves the synthesis of calcium
carbonate crystals at ambient temperatures in calcium rich systems. Biocementation occurs as active
microbes diffuse through the cracks and micro-pits generating calcitic deposits owing to their metabolic
pathway. The calcifying bacterial cultures produce urease or carbonic anhydrase enzyme which is capable
of precipitating calcium in the surrounding micro environment as CaCO3. The bacterial degradation of
urea locally increases the pH and stimulates the microbial deposition of carbonate. The calcium carbonate
produced binds the soil particles together, thus cementing and clogging the grains, and hence improves the
strength and reduces the hydraulic conductivity of the unconsolidated sands. Moreover, these precipitated
crystals can thus fill the cracks and enhance the durability of cement, mortar, and concrete. Incorporating
calcifying bacteria is the essence of developing a self-healing material or "bio-cementing" technology as
bacteria behaves as a long-lasting healing agent.
The calcifying microbes can be isolated from different sources like water springs, soil, ocean,
environments with high pH values or the cement itself. The purified strains can be grown for a 24-hour
period in the laboratory and then blended with the cement or other materials depending on the desired
application. A cheap carbon source like glycerol/molasses is supplemented to the mixture triggering fast
bacterial multiplication. It was found that after the curing time of 28 days, tensile strength, micro-crack
healing capacity, and durability increased significantly. The process is as simple as mixing bacteria into a
cement paste. The technique for creating a high strength cement in a permeable starting material involves
combining the starting material with effective amounts of (1) a urease producing micro-organism with a
high urea hydrolysis rate; (2) urea; and (3) calcium ions, under standard conditions of 0.5-50 mM urea
hydrolyzed min-1. Scientists found that after injecting the bacterial cementitious solution for a period of one
month, the spores of three particular bacteria where still viable. Harmless bacteria such as Bacillus genus
remains dormant until water enters the cracks. In this case, formation water, or water from fracturing fluids
or any source can be used to trigger the bacteria. Moreover, the process does not require oxygenation.
2 SPE-191778-MS

In this paper, self-healing approaches based on bacteria will be thoroughly reviewed. The concept of
biomineralization, bioclogging, and biorepair and its applications in improving the engineering properties of
sands and cement is tackled. Based on the aforementioned aspects of self-healing in cementitious materials,
recommendations for further research in self-healing engineering applications are proposed. This method is
a green and eco-friendly way and the use of bacteria can lead to substantial savings. The following presents
major practical applications for the oil and gas industry. Via the microbial calcification theory, solidifying
the sea beds before drilling for oil, preventing hole cavings and wellbore enlargements or washouts, sealing
undesirable leakage pathways near wellbores to achieve fracture plugging and permeability reduction,
plugging sands to diminish water absorption and porosity are all lucrative potential applications the industry
is in dire need of.

Introduction
Cement is of paramount importance to successful drilling, completion, and production of oil and gas wells.
Cement decreases the surrounding rock-induced load endured by the casing, improves the stress field
distribution, and casing collapse specification. It plays a principal role in providing structural support to the
entire well, prevents casing shoe leakage, and maintains wellbore integrity. It provides zonal isolation and
prevents gas migration which leads to sustained casing pressure (SCP), thus, isolating fluids and restricting
gas flow behind the casing string. However, cement fails due to a variety of reasons as it has to cope
with different stress variations, pressure, and temperature during the lifetime of a well. Recurrent and
dramatic fluctuations of stress, temperature and pressure can cause damage within cement and its interfaces
as well, which increases its probability of damage. Cracking, void formation, and channeling have led to
severe consequences in the oil and gas industry. Squeeze cementing restoration might not be suitable in
situations where the leakage pathways are narrow and inaccessible. Owing to fluid or gas migration, 1 in 3
of primary cementing jobs fail. In fact, in the United States alone, a commonly recited industry number is
15% failure. ‘First-attempt’ cementing squeeze jobs have a comparatively low rate of success. More often
than desired, a squeeze operation would necessitate 2 or 3 unsuccessful attempts before an effective seal
is constructed in the targeted location. In the New Hope Field in the US, it was estimated that a factor
of 5.33 was the average number of squeeze cementing job attempts before success was obtained. Despite
this number, successful isolation was not achieved in all the wells. In the US, remedial jobs repairing
cementing failure related challenges have been estimated to cost more than $50M per year (Cavanagh et al.
2007). For every abandonment/squeeze job per leaking well, the financial burden would cost approximately
$30,000 in manpower, rig rate, material, and transportation which eventually would lead to a compounded
abandonment/repair/loss surpassing $100,000. Contrary to microbial methods, which possess the advantage
of being microscopic, roughly 2μm in size. This can be achieved via fluids of low viscosity. As a result,
aqueous suspensions can directly foster microbially-generated precipitation of minerals. These fluids can
seal small gap fracture leaks or interface delamination.
The second obstacle is drilling in unconsolidated sands. In fact, several casing failures have been reported
due to wellbore and reservoir cavings. Numerical modelling showed that cavities and non-uniform stress
distribution in unconsolidated formations caused most of the failures in the Gudao reservoir of the Shengli
Oilfield, China. The failures appeared to be casing buckling and fracturing (Peng et al. 2007). Poorly
consolidated or unconsolidated sands with lightly cemented or un-bonded grains, constitute the formation
of major fields in the Bohai Bay of China, Gulf of Mexico, and the Campos basin of offshore Brazil (Monus
et al. 1992). These fragile formations adversely affect wellbore stability during drilling and thus increase the
likelihood of casing failure and sand production throughout production. Sand production through erosion
causes formidable casing damage in unconsolidated pay zones in Canadian heavy oil sand (Wagg et al.
1999). Moreover, Morita et al. 1995) argued that the creation of cavities by sand production behind a casing
is one factor that is often overlooked as a cause of casing failure.
SPE-191778-MS 3

Construction Microbial Biotechnology is a novel interdisciplinary approach combining both engineering

and science disciplines. It involves the application of microbially-mediated procedures and microbial
manufacture of construction supplies. Cheap, sustainable, and environmentally-friendly polysaccharides
and cements are beneficial in construction and earth sciences. The activity of diverse microorganisms such
as iron- and sulfate-reducing bacteria, acidogenic, urease-producing, halophilic, alkaliphilic, denitrifying,
cyanobacteria, algae, and microscopic fungi is what construction biotechnology is dependent on.
Processes such as concrete surface biorepair, biocementation, soft clay bioencapsulation, soil biogrouting,
bioaggregation, and bioclogging are some of the applications that can be exploited in the oil and gas industry.
Cemented cracking and channeling permit the seepage of water along with induced-chemical and microbe
damage to penetrate through and damage the existing cement, allowing structural strength reduction and
costly repairs.
In current years, the innovative idea of self-healing cracks has grabbed worldwide interest. Robust
limestone is generated as a result of bacterial metabolic reactions. This process is naturally occurring and
induces crack healing. Unconsolidated sand and soil particles form cementitious links as calcite particles and
crystals deposit on the surface of the static microbial cell. The cement-grain bonded complex forms a solid
mass. To ensure effective bonding, uniform carbonate deposition must be precipitated intergranularly. The
substrate urea and the source of energy in the form of calcium is hydrolyzed by the bacterial enzyme urease
to formulate biocement. These ingredients must coexist for successful production. The chief task of the
microbes lies in their capacity to generate an alkaline surrounding through high pH values and DIC elevation
produced by their physiological biochemistry (Ariyanti et al. 2011). Furthermore, bacterial cells perform as
nucleation sites. They control the precipitation of carbonates through the variation of the precipitation factors
explained above, either independently or in a group pattern (Hammes and Verstraete 2002). The generated
calcite can be exploited as an individual cementing material or in conjunction with other cementitious
solutions for various applications. Self-healing induced blockage of cracks impedes aggressive materials
from entering existing pathways. The process proliferates abrasion amongst the surfaces of the cracks,
consequently leading to soil and rock solidification which elevates its strength and stiffness (Whiffin et al.
2007). This process is illustrated in the schematic below as shown in Fig. 1.

Figure 1—A schematic showing the Crack healing process caused by Bacteria.

Scientists speculate that the generated bio-minerals are both an uncontrolled and unintended by-product
of bacterial metabolic reactions. Biocement mixtures can be provided in a multitude of arrangements. This
includes a liquid form, tubular capsules, or lyophilized powder (Achal et al. 2009a; De Belie and De Muynck
2009; Wang et al. 2012). The microbial solution can remedy present structures and materials via coating,
dipping, or in a spray form (De Belie and De Muynck 2009; Ghosh et al. 2005). Biocement generated MICP
is deposited in the intergranular voids as shown Fig. 2 (Achal et al. 2013).
4 SPE-191778-MS

Figure 2—ESEM imaging displaying the self-healing outcome of bacterially-induced carbonate precipitation on
crack surfaces in concrete samples.Scale bars: left picture 100 μm, right picture 50 μm (Van der Zwaag 2010).

Cracks less than 0.2 mm are self-restored using concrete itself. Conversely, concrete fails to self-heal and
repair cracks greater than 0.2 mm, and that is where bacterial action comes to play using calcite deposition.
(Luo et al. 2015). Crack formation activates bacterial cells from its hibernation state, bacteria metabolizes
the substrates and precipitates calcite and as a result the cracks plug. As soon as the pathways are fully
packed, it triggers the bacteria into its original hibernation mode. It was noticed that bacterial-restored
concrete is stronger compared to traditional concrete.
Several laboratory and field trials were conducted on self-healing agents and methods of evaluation on
the effectiveness of self-healing. (Li and Herbert 2012; Wu et al. 2012; Van Tittelboom and De Belie 2013;
Mihashi and Nishiwaki 2012). A flow tree showing the different approaches to the progression of self-
healing cement-based material is illustrated below in Fig.3. The chart explains the differences between
natural and man-made processes. In this study we exploit the naturally occurring reactions of bacteria using
artificial methods of bacteria induction and material supplementation.
SPE-191778-MS 5

Figure 3—A flow chart of the available cement based self-healing research (Muhammad et al. 2016).

Limestone precipitation by various strains of anaerobic bacteria was initially highlighted in literature
by 1998. In vitro generation of calcium carbonate in cementing applications has paved a pathway for the
advent of MICP technology (Zhu and Dittrich, 2016). Urease mineralization aided in the comprehensive
repair of a calcite surface as supported by Dick et al. (2006). He found out that the water absorption capacity
of limestone was substantially reduced after remediation. Coating monument surfaces with maintenance
mixtures is another application of the healing and sealing activity of microbes. Owing to its synthesis at
ambient temperatures, better sustainability is offered in comparison to traditional approaches. As a result,
it is becoming an extensively acceptable means of bacterial-based cement synthesis. Soil strengthening
and consolidation is another application due to the ubiquity of bacteria in soil and rock (approximately
1012 microbes per kg of soil). Given its ambient temperature manufacturing conditions, it is a lucrative
opportunity for In Situ cementation practices (De Muynck et al. 2010; Dejong et al. 2013; Dhami et al.
2016b). Another instance was in preserving the Angera Cathedral, in Italy using MICP (Perito et al. 2014).
A ground improvement encompassing 100m3 of sand has been rectified via biomineralisation (Van Paassen
et al. 2010). Meanwhile, Wiktor and Jonkers 2015 described a substantial crack sealing event via MICP in a
parking garage. The process of MICP relies on six main aspects: (1) the concentration of calcium, (2) urease
6 SPE-191778-MS

enzyme, (3) carbonic anhydrase enzyme, (4) dissolved inorganic carbon concentration (DIC), (5) pH, and
(6) nucleation sites’ obtainability. There exist two main predominant practices in which MICP has been
performed. The first is biocementation acting as an individual cementing material, whereas, the second is
biodeposition acting as a surface layer (Achal and Mukherjee 2015; Hammes et al. 2003). Biocementation
can be accomplished as bulk via completely supplying the microbial culture solution or by the separate
injection with urea and calcium reagents along with surface filtration. These benefits were advocated by
Reddy et al. 2015: (1) producing biocement takes a remarkably shorter time compared to the conventional
methods and they are synthesized at much lower temperatures. Cement uses a temperature of up to 1500°C in
production. (2) According to experiments, a 38% compressive strength proliferation was noticed in mortar
(3) A curing period of 28 days is sufficient for crack remediation and strength restoration of construction
materials. (4) Inexpensive nutrient sources such as "lactose mother liquor and corn steep liquor" can be
used for bacterial growth. (5) Compared to ordinary cement, biocement boosts sand column durability
by increasing its compressive strength and diminishing its permeability. Finally, (6) it is ecofriendly as
it consumes less energy and produces less CO2emissions. The following chart shown in Fig. 4 displays
the biotechnological process of calcium- and urea based biocement process via microorganism activity.
The progression is divided into three main stages:(1) upstream processes such as reagents preparation and
development of cement, mortar, or concrete for treatment purposes, equipment and microbial suspension;
(2) fundamental practices such as biotreatment, bioclogging, and biorepair techniques; (3) downstream
processes such as decontamination or the disposal of solid, liquid, and gas phases. Microbial growth
and sand grain particle treatment can be amalgamated together or individually by blending the chemical
formulations fabricated as slurry, solution, or suspension forms.

Figure 4—A flow chart displaying the down-stream processes starting with the mixture of the
components all the way to reaching the final "Dry bio-cement" product (Ivanov et al. 2015).
SPE-191778-MS 7

This novel approach of plugging and bio-healing of bacterially-induced concrete micro-cracks is used in a
variety of fields ranging from restoration of construction materials, oil recovery, CO2 sequestration to metal
remediation (De Muynck et al. 2010; Dejong et al. 2013; Dhami et al. 2014a). However, its application in the
oil and gas cement repair and sand unconsolidation issues have remained untapped. As a result, highlighting
this cost-effective environmentally friendly way to repair cement and strengthen the formation to the oil
and gas industry was the motivation behind this review.

Biological Background
Geological establishments such as rocks, sediments, and aquifers result from metabolic consequence of
many microorgansims (Dupraz, C. et al. 2009; Rusznyak et al. 2012; Zhu and Dittrich 2016). Various
processes contributed to redox reaction conditions accountable for calcite deposition in a variety of natural
environments (Dupraz et al. 2009). Amongst these processes include: ammonification, methane oxidation,
denitrification, photosynthesis, ureolysis as well as extracellular polymeric constituents.
Limestone precipitation in natural environments, due to pH increase or DIC, is generated by a wide variety
of diverse bacterial classification (Dhami et al. 2013c; Zhu and Dittrich 2016). These include heterotrophic
and autotrophic classes such as methanogenic bacteria, nitrate reducing bacteria, sulfate reducing bacteria,
cyanobacteria, myxobacteria, and ureolytic bacteria. The vast majority of biocementation work utilizes
Gram-positive bacterial cultures such as Sporosarcina pasteurii particularly S. pasteurii ATCC 11859 strain
owing to its active urease activity and capacity to greatly multiply at pH greater than 8.5 and at elevated
calcium concentrations of at least at 0.75 M.
Terrestrial environments are directed by ureolysis and photosynthetic conduits, whereas, sulfate reduction
accompanied by photosynthesis is governed by fresh water and marine surroundings. The bacterially-
induced mineralization of carbonates in natural settings has been successfully attempted by many
researchers (Dupraz et al. 2009; Dupraz et al. 2009; Rusznyak et al. 2012). Amongst the wide-array of
metabolic routes discussed, ureolytic and carbonic anhydrase courses have been extensively tested for
applications in civil engineering (Smith and Ferry 2000; De Muynck et al. 2010; Qian et al. 2015; Dhami
et al. 2016a).
Ureolytic populations control pH and DIC levels via the hydrolysis of urea. The process initiates when
urease hydrolyses urea to produce carbonate (Eq. 1) which hydrolyses extemporaneously to carbonic acid
and ammonia (Eq. 2). In the presence of water, these yields reach equilibrium producing bicarbonate wherein
hydroxide and ammonium ions are generated causing a pH rise (Eqs. 3 and 4). In alkaline conditions and
calcium presence, these reactions lay the path for calcium carbonate induction (Eq. 5) (Stocks-Fischer et
al. 1999).
(1)
(2)
(3)
(4)
(5)
Carbonic anhydrase (CA) is an abundantly-dispersed alternative enzyme significant in CO2 capture in
carbonate form (Tripp et al. 2001; Dhami et al. 2014b). This enzyme behaves as a prospective catalyst for
CO2 hydration leading to calcite production in the existence of a calcium source (Eqs. 6 and 7):
(6)
(7)
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Organic acids oxidation is an alternative metabolic pathway for the production of calcium carbonate.
It is more environmentally friendly compared to urea hydrolysis which generates excessive amounts
of ammonium (Jonkers 2008). Furthermore, using this metabolic route generates CO2which reacts with
portlandite (Ca(OH)2). This compound serves as an imperative hydration produce of Portland cement to
create more carbonate. (Jonkers et al. 2010; Wiktor and Jonkers 2011).
Bacteria lifetime can be critically extended if it is immobilized in porous clay aggregates prior to concrete
mixing. It was observed that when clay particles intersected cracks, the self-healing process was elicited
(Jonkers et al. 2010; Wiktor and Jonkers 2011; Jonkers and Schlangen 2009; Stuckrath et al. 2014). Attempts
at bacteria immobilization in silica gel or polyurethane have been conducted (Wang et al. 2012). This
is a process where the bacterial cells are condensed in polymers. To shield bacterial spores, Wang et al.
2011 attempted to use agar and sodium alginate. Bacterial nutrients such as organic acids or urea must be
interlocked in the matrix structure (Ren and Qian 2014). It is noteworthy to mention that the presence of
water inside cracks is crucial for bacteria-influenced self-healing. An illustration is shown in Fig. 5.

Figure 5—A graphic representation of a crack-healing process by concrete-immobilized bacteria. The bacteria induced
on the surface of the crack activates by water influx, multiply, and deposit calcium carbonate to plug the cracks and
voids. Consequently, it strengthens and protects the steel from additional harsh exterior chemicals (Jonkers et al. 2008).

Madigan et al. 2008 concluded that bacteria has the capability of surviving a very wide range of acidic
and saline environments and tolerate extreme pressures in addition to severe hot and cold weather conditions
from beneath freezing to above boiling points. Since the beginning of time, bacteria have always been
earth inhabitants, as a result, they do not impose any environmental risks (Fritzges 2005). Urease enzyme
is produced by a significant number of bacterial strains exploited in bio-mediated sand improvement
techniques. These include Spoloactobacilus, Desulfotomaculum, Bacillus, Clostridium, and Sporosarcina.
To assess the influence of further environmental conditions: such as calcium and urea ion concentration
as well as bacterial cell concentration, Okwadha and Li 2010 studied their effects at constant temperature
on Sporosarcina pasteurii. It was discovered that an increase in the bacterial cell concentration is induced
by an increase in urea hydrolysis rate. Moreover, when the calcium ions were multiplied by ten-fold, a
SPE-191778-MS 9

remarkable escalation in calcite precipitates of 100% was documented. Scientists testified that the process
of ureolysis relied greatly on temperature and a range of 20°-37°C optimized MICP. The following was
naturally dependent on the surrounding environment and reactant concentrations in the system. Bacteria -
mediated calcium carbonate formation in accordance to their metabolic routes is given in Table 1 (Vijay
et al. 2017 b).

Table 1—Diverse metabolic routes of bacterial calcite precipitation (De Belie and Wang 2016).

Scientists have speculated that MICP can be explained using one of the three proposed mechanisms.
Primarily, the biomineralization process occurs accidently and unintentionally due to bacterial metabolic
reactions (Knorre and Krumbein 2000). It is safe to assume that this mechanism is the most acclaimed of
all three mechanisms. The mechanism is illustrated in Fig. 6 below. Secondly, although the mechanisms
are not fully understood, there is a wide consensus that ion exchange occurs through bacterial membranes
where the site of carbonate nucleation takes place specifically on the cell wall surface as shown in (Eq.
8). The negatively charged wall attracts Ca2+ present in the surroundings, which then reacts with CO32-
ions. This is where CaCO3 forms and precipitates (Castanier et al. 2000) as shown in (Eq. 9). With
regards to the surrounding formation, a decrease in the negative surface charge occurs with a pore fluid
concentration and valence ion increase (Chapelle 2001). The third mechanism hypothesizes the involvement
of macromolecules located posterior to the bacterial cell in the entrapment of calcium ions (Braissant et al.
2003). Those molecules can also regulate crystallization by behaving as "growth modifiers".
(8)
(9)

Figure 6—A detailed microbial carbonate precipitation progression by the hydrolysis of urea (DeJong et al. 2010).
10 SPE-191778-MS

Methodology
Materials
Depending on the process of choice, whether it is biocementation, biogrouting, or biorepair, different
conditions will be applied. These various conditions include: different microbial solution concentration, type
of bacteria, specimen preparation control solution specification, water salinity, substrate type and quantity,
incubation periods, stoichiometry of chemical reactions, cement type, water to cement ratio, and methods of
testing, analysis, and verification. Sufficient substrate, calcium source, and carbon source should be supplied
for the bacteria to perform the calcite formation reactions. Some of the more common ingredients used
in microbial solution preparation include: soil as a bacterial source, lentil seed powder used as a protein
source, beef extract as a vitamin source, sugar as a carbon source, gypsum as a calcium source and urea as
the substrate (Charpe et al. 2017). Table 2 shows a summary of the various media experimented with for the
microbially-induced precipitation of calcite. The pH of the microbial solution varies at various stages of the
process. Before incubation the pH ranges from 6.8 to 7.2 while during incubation, the process is maintained
from 4.7-5.2. After adding urea and gypsum, it becomes alkaline and the pH values range from 8.5 to 9.5.
While ordinary Portland cement is mainly used in most of laboratory experiments.

Table 2—Summary of the various media experimented with for the microbially-induced precipitation of calcite (De Muynck et al. 2010).

Mortar is composed of both cement and sand while concrete is composed of cement, sand, and gravel
or other fine and coarse aggregate. The addition of water activates the cement, which is the constituent
accountable for creating the mixture and solidifying it onto a mass like structure. Cement can be defined as
the binding agent in both concrete and mortar. It is a mixture of clay, silica sand, limestone, and shells. It is
noteworthy to mention that the failure properties of cement and mortar and the porosity and permeability
characteristics are very comparable. Accordingly, the methods used in this study are all applicable to cement
as a stand-alone fluid.

Methods
Four different methods relatable to the oil and gas industry are defined and proposed as follows:
1. Biocementation: This process is known for its ability to improve sands and soil via microbially-
induced particle-binding constituents.
2. Bioaggregation: particle bioaggregation of is a technique used to aggregate or augment fine particle
size.
SPE-191778-MS 11

3. Soil Biogrouting: Grout, composed of inorganic or organic constituents, behave as dispersants. They
bind materials to enhance their resilience, and they are added to develop materials impermeability
and elevate their mechanical properties. This is done by plugging cavities, cracks or voids and
consolidating sands in cementitious materials or soils. However, chemical grouts can be costly and
toxic as they are constructed with lignosulfonates, acrylamides, and polyurethane. Biogrout, on the
other hand, is formulated via biomineralization. The process exploits calcite induced by bacterial cells.
4. Bioclogging: is a process that reduces soil and porous rock hydraulic conductivity. This is achieved
by inter-pore filling of microbial-produced materials.

Applications
There are several approaches to self-healing, five of the major approaches are listed in Table 3 below.
Supplementary cementing material and bacterial immobilization approaches are focused upon in this review.

Table 3—Self-healing approaches and their measured variables (Muhammad et al. 2016).

As a Cement Admixture
To regulate cement’s excessive "embodied energy", a few calcareous and siliceous by-products such as blast
furnace slag, rice husk, and fly ash have been added to cement. An industrial byproduct should be used
for efficiency purposes along with a locally accessible material. Rice husk ash is a very common blend;
however, it tends to be a contaminant (Dhami et al. 2012). Similarly, microbial products can be exploited as a
cement admixture or in combination with different admixtures. Studies conducted on the addition of fly ash
as an admixture in the biocementation process are still in their infancy (Dhami et al. 2012, 2013a). However,
it was demonstrated that fly ash enhanced the durability of such construction constituents. In an attempt
to cultivate effectual formulations using fly ash as a cement admixture, moisture and temperature levels
were enhanced to optimize bacterial cell life span. A survival span of a twelve months was noticed when
a quantity of 250 g of fly ash was added in to a bacterial solution composed of Lysinibacillus fusiformis
SS18, B. megaterium SS3, and B. cereus SS5. In comparison to the control specimens, calcite precipitation
in sand plugs in the case of concocted bacterial cells resulted in a 24% and 28% decrease in porosity and
water absorption respectively (Dhami et al. 2013a).

Oil-Reservoir Sealing and Permeability Reduction

Bacterial cells were used to plug the highly permeable rocks of the oil-reservoir. High permeability regions
are characterized in some oil-reservoirs. Therefore, during waterflooding, water moves through the path of
highest permeability zones by-passing the desired oil. Owing to the microscopic bacterial size, they travel to
highly permeable zones, sealing the pores and, thus, increasing oil recovery and sweep efficiency by a 100%
(Bryant 1987; Behlülgil and Mehmetoğlu 2002; Knapp et al. 1985). The following should not be mixed
with the traditional microbial enhancement techniques, as pore plugging was the result of microbial growth
and reproduction (MacLeod et al. 1988), and pressure increase due to gas generation. However, residual
oil recovery improvement due to the bacterial hydrolysis of urea proposes a viable substitute for formation
pore blockage. A case in point was offered by (Gollapudi et al. 1995). The authors attained total plugging
12 SPE-191778-MS

in a few days by blending sand and bacteria, followed by a biocementation reagent with carbonate. As
mentioned earlier, it was discovered that the bacteria impacted the CaCO3 precipitation rate along with the
achievable saturation, thus, governing the polymorphic crystal structure of the generated calcium carbonate
(Bosak et al. 2004).
In 2014, Cunningham et al. reported that a MICP-based field demonstration was successfully executed.
The treatment was performed 341 m beneath ground level at the Gorgas 1 well in Walker County, Alabama.
The demonstration used traditional oil and gas equipment including: wireline, a dump bailer, tubing, and
packers. Injection pressure increased until a horizontal fracture was induced in the Fayette Sandstone
formation. Dump bailer delivered the growth nutrients, calcium, urea, and the S. pasteurii strains into the
fractured zone. Calcium with urea solutions (99 g/L and 56 g/L respectively) along with 6 bacterial cultures
(3 × 107 cfu/ml concentration of cells) were delivered in 24 bailer batches over a four-days period. Sealing
and well bore solidification was accomplished. This was observed from a trend of reduced fluid injectivity
and pressure decline.

Biocementation
Biocementation is a technology targeted at sand consolidation. Scientists have proven that biocementation
was effective in aggregating sand particles to increase the compressive strength of bio-sands and other
cementitious materials. It reduces water conductivity and rate of water flow in soil (Rong et al. 2012; Achal
et al. 2009). Bacterial activities produce insoluble inorganic and organic materials, known as biocement,
that contribute selectively to the cementation process. This "green material" is composed of an ionic calcium
solution, an alkalophilic microbe, and a substrate aqueous solution (Achal et al., 2013; Rong and Qian,
2012a) The characteristics of the various biocementation processes are shown in Table 4 (De Muynck et
al. 2010).
Permeability preservation tolerates the application of several treatments, pumping reduced injection
pressures, and treating large areas of sand or soil. Several microscopic procedures assessed the pore volume
space alteration due to calcite deposition; a pore space spatial distribution was observed amongst the
sand grains. This distribution helped in sand stiffness and permeability reduction of the treated specimens
(DeJong et al. 2010). Accordingly, a biocementation In Situ treatment can be performed bottomhole without
the perturbation of existing formation.

Table 4—Characteristics of the various biocementation processes (De Muynck et al. 2010).
SPE-191778-MS 13

MICP process was shown to deposit significant quantities of calcite in subsurface fractures and paths
of least resistance when properly controlled. It has been recommended for several subsurface engineering
applications. Table 5 describes literature-reported reaction conditions of calcite generation by means of
bacterial hydrolysis of urea for several applications highlighting bioclogging and biocementation. Various
successful applications were noted in gas leak prevention for geologic fluid and CO2storage, enhancing
wellbore integrity and unbalanced porous media, and fracture stabilization to name a few (Phillips et
al. 2012; Cunningham et al. 2014; Whiffin et al. 2007). Sealing leakages in water retaining structures
by bioclogging and biosealing applications showed promising results in many civil engineering practices
(Ivanov and Chu, 2008; van Paassen, 2011). A permeability reduction of up to five folds under ambient and
subsurface pressure conditions was noted in both experimental testing and simulation modeling.

Table 5—Various applications of biocementation on sand, cement, and mortar materials and their benefits (Achal et al. 2015).

Cases of reduction in permeability in several 1-inch diameter Berea sandstone cores were noted
using laboratory-based MICP process. Fractures in shale cores and a hydraulically fractured 29-inch
diameter sandstone core were sealed (Phillips et al. 2012; Cunningham et al. 2014). Moreover, Soon et al.
(2014)discovered in his study that shear strength was substantially improved and a consequent decrease
in hydraulic conductivity (69% and 90%, respectively) were verified after a treatment of 48 hours of 0.5
M cementitious solution and a bacterial concentration of 1 x108 cfu/mL. Analogously, current experiments
by Soon et al. (2013) exposed a 96% shear strength improvement on tropical residual soil and sand using
a biocementation mix of 0.5 M concentration. In a study simulating field conditions, MICP was applied
on a 5 m sand column.Following the treatment, an extensive stiffness, load bearing capacity, and strength
advancement was noticed without altering soil permeability. The authors discovered that at a minute urease
concentration and a temperature array of 20 to 50°C boosted CaCO3generation (Whiffin et al. 2007). Sand
experimentation via biocementation was studied in numerous occasions (Achal et al. 2009a; Qian et al.
2010; Rong et al. 2012). Fig. 7 shows an image using the Light Microscope for calcium carbonate crystals
generated via ureolytic bacteria binding of two sand grains. The process involved an experimental setup
composed of a long cylinder filled with layers of sand mixed with bacterial solution. The authors noticed
that the sand turned into a solidified mass after several days (Achal et al. 2009b). CaCO3 precipitates in
the bio-sand totaled to 24% of the total mass of the sand treated with S. pasteurii bacteria. The experiment
was then enhanced with a mutated strain of the bacterial type which gave a 33% improvement (Achal et
al. 2011b). Bio-sandstone strength was found to be 6.1 MPa (Rong et al. 2012). A 25% enhancement in
compressive strength was noticed after blending Shewanella sp. strain, extracted from a hot spring, with
mortar in a biocementation process (Ghosh et al. 2005). Achal et al. 2011 designed mortars implementing
biocementation in combination with traditional Portland cement. A ratio 0.47 of microbes-to-cement was
formulated, similarly for concrete specimens except for the addition of coarse aggregates (Achal et al.
14 SPE-191778-MS

2011). The result displayed a 17–36% increase in compressive strength (Achal et al. 2009; Achal et al.
2011). Additionally, a four-fold improvement of the resistance to water permeability was noticed (Achal
and Reddy 2011). Cheng et al. 2013 studied the outcome of various saturation degrees on the geotechnical
attributes of bio-cemented coarse-grained sands. The authors found that the metabolic processes of aerobes
were optimized at around 60%-80% degree of saturation of cementation solutions. The study suggested
that biocementation was governed by two major factors: greater urease activity which brought about higher
calcite precipitation capacity and denser nutritional compositions trigger calcite deposition. The anaerobic
bacterium, Desulfovibrio desulfuricans was extracted producing calcite, as described in (Eq. 10) (Gauri and
Bandyopadhyay 1999).
(10)

Figure 7—An image using the Light microscopic for calcium carbonate crystals
generated via ureolytic bacteria binding of two sand grains (Al-Thawadi, 2008).

Biogrouting
Ground permeability control and fortification with microbially-generated cementing binding agents was
successfully implemented using biogrout (Whiffin et al. 2007; Ivanov and Chu 2008; DeJong et al. 2010). A
prevailing part of literature on the development of biogrout is founded on MICP. The process is analogous
to MICP biocementation, bioclogging, and biorepair. The calcitic crystals form interlinks between the sand
particles to seal cracks as bacteria multiplies in the existence of water or any kind of moisture and oxygen
in case of aerobic bacteria (Achal and Mukherjee 2015). The solid carbonate reduces sand porosity and
permeability values. Similarly, urease-based biogrout remedies cementitious materials and modifies sand
consolidation factors (Whiffin et al. 2007; DeJong et al. 2010; Rong et al. 2012). Using real-life techniques
and field conditions, scaled up experimentations verified the theoretical feasibility of using biogrout for
formation improvement. This can be achieved through injection at a single region or throughout a horizontal
range using screens of production and injection wells where a substantial variation of 0 to 12 MPa is noticed
in unconfined compressive strength (UCS). Cheng and Cord-Ruwisch 2012 found that bigrouting a one-
meter sand columnresulted in the formation of homogenous cementitious precipitates. The formed calcite
crystals yielded great strength which was partly due to the reduced water amount in the associated pore
space. In another experiment, MICP-induced biogrouting was used on poorly graded coarse sand heaps
to ameliorate its mechanical properties. At 30 °C temperature, 21.45 mmol/min of S. pasteurii bacteria
was cultured in an ammonium-yeast medium using a buffering solution (Mahawish et al. 2016). The
treatment procedure consisted of two biochemical phases which were replicated sixteen times. It consisted
SPE-191778-MS 15

of consecutive infiltration of cementation solutions and bacterial formulations (Cheng 2012). To quantify
the volume of calcium carbonate deposition, a hydrochloric acid washing procedure was implemented.
Calcite was noted as white precipitations on coarse sand surfaces approaching 21%– 22% based on dry
weight, producing significant strength and stiffness enhancements. These findings pave the way for bigrout
application in situ sandstone formations.

Bio-Restoration
In 2016, a bio-restoration method by Chen et al was executed to rapidly enhance the self-healing capacity
of cracks that were mainly cement-based. The nutrients were a mixture of calcium nitrate, sucrose, and
Brewer’s yeast extract. A water to cement ratio of 0.42 was used to make the samples. At a 30 ± 2 °C
temperature, the bacterially-treated cracked samples were fully submerged in tap water. The substrate and
nutrients were uniformly mixed and immobilized on a carrier, while bacteria (Bacillus mucilaginosus)
was immobilized using a Ceramsite carrier. Following a 21day curing period, sizeable quantities of white
precipitates were noticed on the surface of the sections and inside the cracks, which was later confirmed
using XRD and SEM/EDS analyses. While, a minor change in the sample apparent water permeation
coefficient was noticed after a 28-day restoration period. The rate of area repair on specimen surfaces with
nutrients and bacteria on the Ceramsite carrier built up to 87.5% while the flexural strength of the restored
samples elevated to 72% from the original 56% compared to other microbiological methods such as the
mineralization of carbonic anhydrase, silica gel, and polyurethane immobilized bacteria.

Conclusions and Future Work

The objective of this paper was to present a comprehensive review on bacterial self-healing capacity
and highlight the importance of using interdisciplinary research in the fields of construction microbial
biotechnology and geotechnical engineering in serving the oil and gas industry. The objective of this paper
was to expose MICP technology and its various techniques to the oil and gas sector and identify its breadth of
opportunity. Although MICP applications in the oil and gas industry is still in its infancy, it has tremendous
potential. Sealing cementing cracks, rectifying the presence of cement voids due to air entrapment and
mitigating channeling, providing zonal isolation, consolidating sands. Other applications include solidifying
seabeds, sealing hydraulic fractures, self-healing of offshore concrete structures, and reducing the potential
of leakage from the subsurface. This sealing technology is cost-effective and environmentally friendly. In
fact, microbial cement could be commercially manufactured with lower cost compared to traditional cement.
Future work would be targeted to evaluate the feasibility of various chemical formulations to foster MICP
and advance optimized injection strategies to promote further MICP field-scale success. In addition to the
assessment of the impact of MICP minerals to supercritical CO2,an integration of both experimental work
with modelling can improve the MICP sealing technology in the subsurface. This permeability reduction
can aid in CO2 sequestration and enhance the storage capacity of underground formation.

Acknowledgements
The author gratefully acknowledges Dr. Ahmed Assem for his advice, help, and guidance.

Nomenclature
DIC dissolved inorganic carbon
EDS energy dispersive spectrometry
ESEM environmental scanning electron microscope
MICP microbially induced calcite precipitation
SCP sustained casing pressure
SEM scanning electron microscopy
16 SPE-191778-MS

UCS unconfined compressive strength

XRD X-ray diffraction

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