ARTICLE IN PRESS

Progress in Energy and Combustion Science 34 (2008) 714– 724

Contents lists available at ScienceDirect

Progress in Energy and Combustion Science

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

Biotechnology in petroleum recovery: The microbial EOR

Ramkrishna Sen 
Department of Biotechnology, Indian Institute of Technology (IIT), Kharagpur, West Bengal 721302, India

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

Article history: Biotechnology has played a significant role in enhancing crude oil recovery from the depleted oil
Received 17 January 2006 reservoirs to solve stagnant petroleum production, after a three-stage recovery process employing
Accepted 5 May 2008 mechanical, physical and chemical methods. Biotechnologically enhanced oil recovery processes, known
Available online 20 June 2008
as microbial enhanced oil recovery (MEOR), involve stimulating indigenous reservoir microbes or
Keywords: injecting specially selected consortia of natural bacteria into the reservoir to produce specific metabolic
Microbial enhanced oil recovery events that lead to improved oil recovery. This also involves flooding with oil recovery agents produced
Mechanisms ex situ by industrial or pilot scale fermentation. This paper essentially reviews the operating
Solvents mechanisms and the progress made in enhanced oil recovery through the use of microbes and their
Gases
metabolic products. Improvement in oil recovery by injecting solvents and gases or by energizing the
Biosurfactants
reservoir microflora to produce them in situ for carbonate rock dissolution and reservoir re-
Biopolymers
Biofilms pressurization has been enunciated. The role of biosurfactants in oil mobilization through emulsifica-
Selective plugging tion and that of biopolymers for selective plugging of oil-depleted zones and for biofilm formation have
SRB been delineated. The spoil sport played by sulfate-reducing bacteria (SRB) in MEOR has also been briefly
Modeling reviewed. The importance of mathematical models used in predicting the applicability of an MEOR
Field trials strategy and the microbial growth and transport has been qualitatively discussed. The results of some
laboratory studies and worldwide field trials applying ex situ and in situ MEOR technologies were
compiled and interpreted. However, the potential of the MEOR technologies has not been fully realized
due to poor yield of the useful microbial metabolic products, growth inhibition by accumulated toxic
metabolites and longer time of incubation. A complete evaluation and assessment of MEOR from an
engineering standpoint based on economics, applicability and performance is required to further
improve the process efficiency for writing more success stories. Thus, this review attempts to address
almost all the issues concerning the MEOR, its past and recent trends and its future prospect and
directions.
& 2008 Elsevier Ltd. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 715
2. Enhanced oil recovery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 715
3. Microbial enhanced oil recovery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 715
3.1. MEOR mechanisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 716
3.1.1. Role of biosurfactants . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 717
3.1.2. Role of biopolymers and biofilms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 718
3.1.3. Use of gases and solvents as MEOR agents. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 719
4. Sulfate-reducing bacteria—Notorious villains in MEOR. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 719
5. Importance of mathematical modeling in MEOR—a qualitative analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 719
6. Field trials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 720
7. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 722
Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 723
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 723

 Tel.: +91 3222283752; fax: +91 3222278707.

E-mail address: rksen@yahoo.com

0360-1285/$ - see front matter & 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.pecs.2008.05.001
 ARTICLE IN PRESS
R. Sen / Progress in Energy and Combustion Science 34 (2008) 714–724
1. Introduction
Crude petroleum is present worldwide in the complex capillary
network of oil reservoirs. Traditional oil recovery technologies
under the umbrella of chemically enhanced oil recovery (CEOR)
can recover a maximum of 40–45% of the oil initially in place, in
two stages, namely, primary and secondary recovery. Thus, these
conventional oil recovery operations often leave two-thirds of the
oil in the reservoir. Primary recovery produces oil and gas using
the natural pressure drive of the reservoir [1–4]. Secondary
recovery involves stimulating the oil wells by the injection of
fluids, which fracture the hydrocarbon-bearing formation to
improve the flow of oil and gas to the wellhead. Several
techniques were used for injecting fluids into an oil reservoir to
augment the natural forces in secondary recovery [4]. When water
is the injection fluid, the process is called water-flooding. Injection
of natural gas, which is an expensive option, constitutes an
operation, called pressure maintenance [4,5]. Other techniques
involving mechanical and physical means such as pumping and
gas lift help in oil production when the reservoir pressure
dissipates. While primary recovery produces 5–10% of the total
reserve, recovery efficiencies in the secondary stage vary from 10%
to 40% of the oil in place [3–5].
To meet the rising energy demand worldwide, there is a dire
need to produce more crude oil. Stagnant oil production and
unimpressive recovery by conventional methods have been a
major concern. The socio-political factors, which have become
increasingly important in the post-Iraq war scenario, are adding
fuel to this fire of concern. During the turbulent times of the Arab
oil embargo, most major oil companies in the USA had set up their
own research centers, and funded major programs to develop new
technologies. These programs resulted in the creation of the
enhanced oil recovery (EOR) technologies, which kept the
industry going strong and competitive /http://www.netl.doe.
gov/scngo/Petroleum/Exploration%20&%20Production/EOR/
eor.htmlS. Hence, attention has been focused on the EOR
techniques for recovering more oil from the existing and
abandoned oilfields. Of these methods, microbial enhanced oil
recovery (MEOR) has the potential to be cost-efficient [1]. MEOR
refers to petroleum recovery methods, which involve the use of a
mixed microbial population and the metabolic products including
biosurfactants, biopolymers, biomass, acids, solvents, gases and
also enzymes to increase recovery of oil from depleted and
marginal reservoirs, thereby extending the life of the oil wells
[1–5]. Critical evaluation of the physical and biochemical
mechanisms that control microbial responses to the hydrocarbon
substrates and their mobility is a prerequisite for a comprehensive
review on MEOR. Such reviews incorporating the in-depth
analysis of the aspects of accession and transformation of
hydrocarbons, changes in cell membrane architecture, cell-surface
adhesion and hydrophobicity, chemotaxis, and also of the
production and quality improvement of petroleum feedstock
and petrochemicals have been presented [1,3–7,14–16]. Thus, the
Improved Oil Recovery Scheme
Thermal Chemical
(i) Steam flood (i) Polymers
(ii) Combustion (ii) Surfactants
(iii) Hot water (iii) Alkali
Fig. 1. Flow sheet diagram showing the process steps in enhanced oil recovery (EOR).
715
purpose of this review is to discuss the operating mechanisms of
MEOR and to highlight the recent developments and future
prospects in this important area of scientific research.
2. Enhanced oil recovery
The flow diagram (Fig. 1) shows the types of EOR processes
that are currently employed in the oil industry. In the US alone the
original oil in place has been estimated to be 650 billion barrels, a
major part of which is the target of improved and advanced oil
recovery methods (Fig. 2). Thus, more advanced technologies are
being implemented in the oil industry today to recover the
trapped oil under the program of EOR. In the US, the EOR methods
including MEOR have the potential to recover much of that
remaining oil, which is estimated to be about 375 billion barrels.
However, oil recovery is challenging because the remaining oil
is often located in regions of the reservoir that are difficult to
access and the oil is held in the pores by capillary pressure [6–8].
No single EOR or MEOR approach can address this problem.
A combinatorial strategy that involves the injection of a fluid or
fluids to supplement the natural pressure in a reservoir, where the
injected fluids interact with the reservoir rock/oil/brine system to
create favorable conditions for maximum oil recovery, could be an
efficient approach for the EOR [7,8]. The effect of capillary forces
on trapping of oil within the pores of reservoir rock is normally
characterized by the use of a dimensionless number, the capillary
number. The capillary number is defined as the ratio of viscous to
capillary forces as given in Eq. (1) [9,10]:
Viscous forces vm
NCap ¼ ¼ , (1)
Capillary forces s cos y
where v and m are the velocity and viscosity, respectively, of the
displacing fluid, s is the oil–water interfacial tension (IFT) and y is
the contact angle. The greater the capillary number, the lower the
residual oil saturation in the reservoir and hence higher the oil
recovery [10]. The normal microscopic view and a cross sectional
view of a typical oil reservoir and the method of water-flooding
are shown in Fig. 3.
3. Microbial enhanced oil recovery
Microorganisms, harmful and helpful, are ubiquitous in
reservoirs around the world. These microbes are increasingly
being acknowledged for their ability to influence reservoir
behavior and oil mobilization [11]. The following are the oil
recovery agents used for CEOR. Surfactants are used to reduce
interfacial tension between oil and water, oil and rock interfaces.
Polymers are used to increase viscosity of water-flood. Acids,
gases and solvents are used to increase the permeability through
the porous network and to re-pressurize the oil reservoir
[1–8,10,12–16]. Since these are petrochemicals, obtained from
Gas Injection Biotechnological Novel
(i) CO2 (i) Seismic / Sonic
(ii) N2 MEOR/MIOR stimulations
(iii) Flue gas (ii) Electromagnetic
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716 R. Sen / Progress in Energy and Combustion Science 34 (2008) 714–724

Not found, but recoverable by

conventional technologies Not found and unrecoverable by
(30 billion barrels) conventional technologies
(37 billion barrels)
Known Reserves (22
billion barrels)

Found, but unrecoverable by conventional

Current cumulative production technologies, target for new EOR technology
(183 billion barrels) (377 billion barrels)

Fig. 2. Original oil in place in the USA (649 billion barrels), showing targets for EOR /http://www.netl.doe.gov/scngo/Petroleum/Exploration%20&%20Production/EOR/
eor.htmlS.

Table 1
Microbial consortia and their metabolites with applications in MEOR

Microbial Example microbes Application in MEOR

product

Biomass Bacillus, Leuconostoc, Selective plugging and wettability

Xanthomonas
Surfactants Acinetobacter, Arthrobacter,
Bacillus, Pseudomonas
Polymers Bacillus, Brevibacterium,
Leuconostoc, Xanthomonas
Solvents Clostridium, Zymomonas,
Klebsiella
Acids Clostridium, Enterobacter,
Mixed acidogens
Gases Clostridium, Enterobacter
Fig. 3. A cross sectional view of an oil reservoir during water-flooding (adapted
Methanobacterium
from http://www.titanoilrecovery.com/titanprocess1.html).
alteration
Emulsification and de-emulsification
through reduction of IFT
Injectivity profile and viscosity
modification, selective plugging
Rock dissolution for better
permeability, oil viscosity reduction
Permeability increase, emulsification
Increased pressure, oil swelling, IFT
and viscosity reduction

petroleum feedstock after refining and downstream processing,

CEOR methods turned out to be economically unattractive as the
finished products are utilized for the recovery of raw materials.
This is why the scientist looked for a cost-effective alternative and
discovered the same in MEOR, which exploits microorganisms for
the production of all the chemicals as mentioned above, albeit at a
slower rate and with lower yields. In fact what is known today as
MEOR finds its history dating back to 1947 [12]. Microbial
consortia, as shown in Table 1, have been used in MEOR. They
have the ability to produce biosurfactants, biopolymers, acids,
gases and solvents to perform the job of recovering residual oil by
fermenting cheaper raw materials, e.g. molasses [1–5,11–18,
21,23–27]. Biomass, which mostly secretes exopolysaccharides
and forms biofilms, aids in MEOR by selective plugging of oil-less
zones [1–4,27–30], as shown in Fig. 4. This has also been studied
by nutrient resuscitation and growth of starved cells in sandstone
cores [20]. Though MEOR is considered as an environmentally
compatible tertiary oil recovery method and is a time tested and
increasingly applied method of oil treatment in the industry as
well, there is also growing application of microorganisms for
the treatment of petroleum-based products both in reservoirs and
on earth’s surface for bioremedial cleanup of hydrocarbons
[1,3–5,13,16,19].
MEOR scores over other EOR processes on two accounts.
Firstly, the microbial cell factories need little input of energy to
produce the MEOR agents and secondly, the application of
microbial processes does not directly depend on the global crude
oil price. In an in situ process, stimulation of the indigenous
microflora by injecting suitable nutrients serves to enhance oil
mobilization. It is the exponential nature of microbial growth,
which leads to the production of useful biochemical agents for
MEOR at higher rates from inexpensive and renewable resources.
However, there are a few shortcomings of MEOR. One major
obstacle that has slowed the implementation of MEOR has been
the difficulty in isolating and/or engineering microbial strains,
which can survive in the extreme environment of the oil
reservoirs. MEOR, to be economically viable, demands the use of
microbial strains, which remain viable at such reservoir condi-
tions as temperatures up to 85 1C, pressure over 17.23 MPa,
extremes of pH and salinity. Genetic engineering tools and
techniques are being used to develop microorganisms that cannot
only survive and grow in extreme reservoir environment, but can
also subsist on inexpensive nutrients and produce substantial
amounts of metabolic products as EOR agents including enzymes.
Genetically engineered MEOR (GEMEOR) and the enzyme en-
hanced oil recovery (EEOR) constitute the advanced MEOR
methods, which are not discussed here.
3.1. MEOR mechanisms
The MEOR processes involve the use of reservoir microorgan-
isms or specially selected natural bacteria to produce specific
metabolic events that lead to enhanced oil recovery. The
mechanisms by which MEOR processes operate can be quite
 ARTICLE IN PRESS
R. Sen / Progress in Energy and Combustion Science 34 (2008) 714–724
Fig. 4. In situ MEOR processes (a) the thief zones are the targets for plugging by biomass or biopolymers, injected with water-flood; (b) flooded nutrients stimulate
indigenous and injected microbes to produce biosurfactants for emulsification (adapted from http://www.titanoilrecovery.com/titanprocess1.html).
complex and may involve multiple biochemical process steps
[1–8,11–16,26]. In selective plugging approaches, microbial cell
mass and/or biopolymers plug high-permeability zones and
lead to a redirection of the water-flood [27–30]. In other
processes, biosurfactants are produced in situ which leads to
increased mobilization of residual oil and reduces oil viscosity
[1,3,5,7,8,11,26,41,45].
In a MEOR process, conditions for microbial metabolism are
supported via injection of nutrients. In some processes, this
involves injecting a fermentable carbohydrate including molasses
into the reservoir [11]. Some reservoirs also require inorganic
nutrients as substrates for cellular growth or for serving as
alternative electron acceptors in place of oxygen. A patented
method involves injecting water containing a source of vitamins,
phosphates and an electron acceptor such as nitrate into the
formation and allowing anaerobic bacteria, either already present
in or introduced into the formation to multiply using the oil as the
main carbon source [25]. The mechanics of MEOR at a molecular
level must be thoroughly understood to assess both the efficiency
and the economic viability of the process. The microbes in MEOR
are mostly hydrocarbon-utilizing, non-pathogenic and are natu-
rally occurring in petroleum reservoirs [3–5,16,22]. It is important
to note that various Bacillus strains can produce the MEOR agents
when grown on glucose mineral salts medium. These strains alone
can be used when oil viscosity reduction is not the primary aim of
the operation. But a consortium of both types of bacteria would be
more useful and preferred. There are two major ways in which
microbes may contribute to EOR: (a) they grow in reservoir rock to
produce gases, biosurfactants, biopolymers and other non-toxic
biochemical to recover trapped oil; and (b) they can selectively
plug high-permeability channels so that the sweep efficiency of
the recovery process can be increased (Fig. 4). It is of utmost
importance to the petroleum industry that these microbial
products should cause a series of very desirable changes in the
physicochemical properties of the crude, and a marked improve-
ment or a near-complete restoration of the lithological properties
of the reservoir rock [3–8,11–17,22–26]. Success of in situ MEOR
operations depends on developing microbial consortia that can
survive and produce the desired metabolic products in reservoirs
containing hydrocarbons and saline water [5,7,14]. Research
activities are continuously focusing on anaerobic extremophiles
including halophiles, barophiles and thermophiles for better
adaptation to reservoir conditions [13–16,32,41,92–94].
3.1.1. Role of biosurfactants
Surfactant EOR represents one of the most promising advanced
methods to recover a substantial proportion of the residual oil
717
[5,7–11,14–16,22,26]. In this technology, an aqueous surfactant
formulation is injected into a mature oil reservoir. Where this
solution contacts the small blobs of oil trapped in the pores of the
reservoir rock, it dramatically reduces the interfacial tension (IFT)
and mobilizes this trapped oil by increasing the capillary number
[7–11,26,31–34]. Formation of an oil-in-water emulsion often
leads to an improvement in the effective mobility ratio until the
surfactant is diluted or otherwise lost due to adsorption on the
rock. The core floods conducted in Berea sandstones using Yates
stocktank oil and synthetic brine showed the ability of these
surfactants to improve wettability for enhanced oil recovery
[9,10]. Surfactants thus contribute positively to improve oil
recovery by reducing IFT and also by altering the wettability of
reservoir rock for water-flood to displace more oil from the
capillary network. The Department of Energy /http://www.netl.
doe.govS sponsored projects at the California Institute of
Technology to design new surfactant systems to make surfactant
EOR process commercially attractive. Another project sponsored
by DOE was to employ advanced biotechnology methods to
enhance biosurfactant production from selected bacterial strains
acclimatized to reservoir conditions. As biosurfactants play a
major role in MEOR [5,7–10,13–16,26,31–41], production and
application aspects of rhamnolipid, sophorolipid, glycolipid and
lipopeptide biosurfactants assume great industrial significance
and thus have been extensively reviewed in literature [42–45].
Biosurfactants with potential applications in MEOR are listed in
Table 2. Though glycolipid biosurfactants produced by Pseudo-
monas strains have been extensively used in MEOR experiments,
lipopeptides like surfactin, lichenysin and lipid polysaccharide
complexes like emulsan have also been found very effective.
Surfactin, a lipopeptide and the most potent microbial surfactant
known so far, has been produced ex situ in the controlled
environment of a fermenter and has been reported to be
successfully used in MEOR [35]. A bioprocess was developed
and optimized for the enhanced surfactin production in three
stages, namely, optimization of the nutritional factors in the
production medium [46], optimization of the environmental
parameters including pH, temperature, rates of agitation and
aeration [47] and optimization of the inoculum age and size for
judicious dosing of seed culture [48]. The concentration and
purification parameters of surfactin when recovered by ultrafil-
tration method in one step after broth clarification have been
characterized and reported recently [49].
Usually, the biosurfactants, which are produced in fermenters
ex situ, are injected with other chemicals in the water-flood to
facilitate the emulsification of oil with the water jet for better
recovery. However, in situ production and application of biosur-
factants in a limestone reservoir has recently been reported [95].
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718 R. Sen / Progress in Energy and Combustion Science 34 (2008) 714–724

Experimental studies have been performed to assess the potential and restore the sweep efficiency of water-flooding operations
of biosurfactants for their applications as MEOR agents [5,7,11–16,21]. Some biopolymers that have potential applications
[22,26,31,32,34,45,50–54]. In order to show the efficacy of some as mobility control agents in MEOR are listed in Table 4.
of the biosurfactant producing strains, the results of laboratory Selective plugging was initially thought to have occurred
scale simulated experiments on emulsification index and percen- primarily due to sieving action which is best achieved when the
tage of oil recovered from sand packed columns are presented in average pore throat radius is less than twice the diameter of the
Table 3. The emulsification index (E24) was determined by adding bacteria [71]. Dead bacteria or non-slime producing bacteria
6 ml crude petroleum to 4 ml culture broth in a graduated tube, either do not plug porous media, or plug the media to a lesser
followed by vortexing at high speed for 2 min. The emulsion extent than do similar strains that produce an exopolysaccharide
stability was determined after 24 h. The emulsification index is layer. The polysaccharides secreted by many strains of bacteria
thus defined as the height of the emulsion layer, divided by the serve mainly to protect the bacteria against desiccation and
total height, multiplied by 100. predation, as well as to assist in adhesion to surfaces
[3,7,13–15,21,65,71]. The effects of biopolymers on fluid conduc-
tivity in sand columns and sandstone permeability have been well
3.1.2. Role of biopolymers and biofilms studied in laboratory trials [17,18,27,30,64–69]. As plugging and
Biopolymers have been used in MEOR experiments mainly for mobility control agents, biopolymers are perhaps more efficient
selective plugging of oil-depleted zones and hence, for perme- than the bacterial bodies. Application of these processes in field
ability modification [3,5,7,11,15,18,20–24,30,57–66]. In selective trials has made the feeding program effective in slowing the
plugging approaches, biopolymers plug high-permeability thief deterioration of hydrocarbon in wells undergoing water-flooding
zones to redirect the water-flood to oil-rich channels. This is an operations by diverting the water jet from high-permeability
adjunct to water-flooding operations, in which water is pumped zones to oil-rich zones [1–8,11–16,23,27,30,64–69,71–72,85–87].
into injection wells in the reservoir in order to force the oil up to The use of bacterially produced polysaccharides as floodwater
the surface bypassing the oil-depleted zones in the reservoir thickening agents is well known. Xanthomonas species has been
[23,27,30,32,57–66]. Bacterial plugging usually involves the feed- used to ferment carbohydrates and produce a thermally stable
ing of injected or in situ bacteria within the reservoir heteropolysaccharide called xanthan gum. Xanthan gum has been
[3,5,7,11,21,23,27,30]. Bacteria and/or nutrients preferentially used in water-flooding systems. Though xanthan gum is an ex situ
enter the reservoir along high-permeability pathways. Biomass product of fermentation of carbohydrates, the use of it comes
growth in those laminae plugs the pore throats, thus decreasing under chemical enhanced oil recovery. The efficacy of this
the permeability in what had once been the high-permeability biopolymer as an oil recovery agent has been studied by injecting
zones. This tends to equalize the permeability across the reservoir an aqueous mixture of the polysaccharide (xanthan gum) and
sodium chloride in a model reservoir in the laboratory [57,68,69].
Table 2 Processes based on the uses of xanthan gum in EOR involve
Various potential microbial surfactants for application in MEOR thickening the injection waters; using the biopolymer with other
Biosurfactant and microbial source Reference
chemicals or adsorption agents to prevent adsorption of the
polysaccharide on to rock surfaces; using multivalent cations to
Source: Bacillus sp. bind the polysaccharide molecules together for preferential
Surfactin Schaller et al. [35] partial plugging and applying with gel particles, like hydrolyzed
Rhamnolipid Banat [26]
polyacrylamides, and ferric chloride solutions to facilitate the
Lichenysin Jenneman et al. [31] thickening process [56,58,85]. Its physical properties of viscosity,
McInerney et al. [33]
Yakimov et al. [34]
Horowitz and Griffin [36]
Table 4
Exopolysaccharide biopolymers used in EOR with their microbial sources [57–66]
Source: Acinetobacter sp.
Emulsan Rubinovitz et al. [37] Biopolymer and microbial sp. Reference
Alasan Navon-venezia et al. [38]
Xanthan gum (Xanthomonous sp.) Pollock and Thorne [59]
Source: Pseudomonas
Pullulan (Aureobasidium sp.) Cho et al. [62]
Rhamnolipid Arino et al. [39]
Levan (Bacillus sp.) Akit et al. [63]
Source: Rhodococcus sp. Curdlan (Alcaligeness sp.) Buller and Vossoughi [64]
Viscosin Neu et al. [40] Dextran (Leuconostoc sp.) Kim and Fogler [65]
Trehaloselipids Tango and Islam [41] Scleroglucan (Sclerotium sp.) Sandford [66]

Table 3
Results of laboratory scale simulation experiments are given

Microbial strain Emulsification index (E24) % Oil recovered from sand packed column References

Bacillus sp. AB-2 80–90 90–100 Banat [50]

Bacillus subtilis MTCC 2423 90 62 Makkar and Cameotra [51]
Bacillus subtilis MTCC 1427 33.3 56 Makkar and Cameotra [52]
Serratia marcescens 94 82 Pruthi and Cameotra [53]
Arthrobacter protophormiae 60 90 Pruthi and Cameotra [54]
Bacillus subtilis DSM 3256 57–63 87–94 Sen (unpublished data) [55]

The glass column that was used for surfactin from B. subtilis DSM 3256 was 36  12.5 cm in dimension and was packed with 100 g acid washed sand. The pH of the column
packing was adjusted to 6.5–6.6 after repeatedly washing with distilled water. The packing was subsequently dried in an oven at 60 1C overnight before being used for the
oil recovery experiment. Crude surfactin (approximately 1 g/L in broth) was used in this study. The results were compared with the literature values.
 ARTICLE IN PRESS
R. Sen / Progress in Energy and Combustion Science 34 (2008) 714–724
Table 5
Solvents, acids and gases with their producing microorganisms and MEOR
mechanisms
Solvents, acids and Operating References
gases with their mechanisms
microbial sources
Acetone, butanol, Improved [12,14,68,71]
propan-2-diol permeability by
(Clostridium, carbonate rock
Zymomonas, dissolution and oil
Klebsiella) viscosity reduction
Propionic and butyric Enhancement of [15,57,68,71]
acids (Clostridium, permeability and
Enterobacter, mixed degree of
acidogens) emulsification
Methane and Rock re-pressurization, [57,68,71,106]
hydrogen oil swelling, reduction
(Clostridium, of IFT and oil viscosity
Enterobacter,
Methanobacterium)
shear resistance, temperature and salt tolerance make it almost an
ideal polymer for use in EOR [59–61]. It is considered superior to
polyacrylamide for the above-stated reasons. However, it is more
expensive and more susceptible to bacterial degradation [67].
Another biopolymer, scleroglucan, is produced by a species of the
fungus Sclerotium and was examined for possible use in EOR
[66,70]. Biopolymer levan was found to be potentially suitable in
oil reservoirs which have a temperature of less than 55 1C, a pH
between 6 and 9, a pressure less than 500 atm and a salt
concentration of 4% [63]. Though xanthan gum has been the most
effective biopolymer, applications of other biopolymers like
curdlan, dextran and pullulan in MEOR have also been reported
[1,3–5,11,62,64–66].
On the other hand, biofilms are heterogeneous systems
of bacteria, their exopolysaccharides and water channels. Studies
on biofilms indicated that they were composed of less than
27% bacterial bodies and the remainder (73–98%) was assumed
to be composed of extracellular products, probably exopolysac-
charides and void space [69,71,72,85]. Biopolymers may occur in
several different morphologies forming biofilms within porous
media like oil reservoirs. Within the porous media, the morphology
may assume a web like structure, called bioweb at a certain stage
of growth [72]. Biopolymer production and consequent biofilm
formation are both important for MEOR operations to be effective.
These two phenomena are influenced by such parameters as
water chemistry, pH, surface charge, microbial physiology,
nutrients and fluid flow [3,66,71].
3.1.3. Use of gases and solvents as MEOR agents
The use of bacterial consortia to produce gases, solvents and
acids for enhancing oil recovery by exploiting the mechanisms of
reservoir re-pressurization and carbonate rock dissolution has
been an old practice [1,12,14,15,68,71,106,114]. The microbial
consortia and the operating mechanisms are enlisted in Table 5.
Bacteria can ferment carbohydrates to produce gases such as CH4,
CO2 and H2. Such gases produced in situ can contribute to
pressure build-up in a pressure-depleted reservoir. These gases
may dissolve in the crude oil and reduce its viscosity. Additional
benefits of this bacteria-induced fermentation process include the
production of acids, such as acetic and propionic acids and the
production of solvents, such as acetone, ethanol, 1-butanol and
butanone. Both gases and solvents can dissolve the carbonate
rock, thereby increasing its permeability and porosity; anaerobic
bacteria, Clostridium acetobutylicum, Enterobacter cloacae and
719
Methanobacterium have been the microbes of choice for their
availability as indigenous natural reservoir microflora. Extensive
research has been carried out in understanding the biofilm growth
in porous media to harness indigenous microorganisms that
promote increased oil recovery from depleted oil reservoirs at low
cost.
4. Sulfate-reducing bacteria—Notorious villains in MEOR
The sulfate-reducing bacteria (SRB), which have relatively
simple growth requirements in sulfate and carbon as energy
sources, play a very negative role in MEOR [11,78]. SRB are
traditionally known to be active in shallow wells, but the
existence of significant SRB populations in deep, hot and high-
pressured reservoirs has been confirmed from souring of the
formations. The association of the onset of reservoir souring,
which jeopardizes the economical calculations, with the starting
of seawater injection in previously so-called sweet fields is largely
due to SRB activities [11,78]. The SRB are extremely sturdy and can
survive prolonged starvation in seawater at both surface and
reservoir temperatures. In this dormant state, starved SRB tend to
be smaller in size than their growing counterparts and may travel
longer distances through the porous network during water-
flooding. Chemicals, such as biocides, are regularly injected as
slug doses in an intermittent treatment program to kill SRB. But
starved SRB are less affected by standard biocide treatments than
actively growing populations [3,5,11,78].
The continuous biogenic sulfide contamination of huge rock
formations and production facilities has belied the theory that
microorganisms cannot significantly influence the micro-environ-
ment of the oil reservoirs [73]. The major concerns of the global oil
industry include souring, corrosion caused by H2S, plugging by iron
sulfide, the related financial burden and the threat to health and
safety of the operators. The so-called Bio-Competitive Exclusion
(BCX) technology /http://www.ptac.org/res/dl/rest0302p.pdfS ex-
ploits this potential and targets indigenous de-nitrifying bacteria
(DNB) and manipulates the entire reservoir microflora to release
trapped oil for commercial production, as well as inhibit the sulfide
production by SRB almost permanently [89]. This oil-mobilizing
and sulfide-removing microbial system was first introduced to
industry as Max-Well 2000, which has proven its efficacy in the
real time field tests involving the injection of low-cost nutrients
[73]. There are reports on the effects of hydrocarbon utilizing SRB
on corrosion of metallic parts of the drilling pipes in injection and
production wells and also on nitrate-mediated microbial control of
reservoir souring on the efficiency of MEOR [74,75]. The dynamics
of microbial control of H2S production in oil reservoirs has been
elucidated [76]. The control of iron and H2S has been tested in an
oil field by employing BCX process [77].
5. Importance of mathematical modeling in
MEOR—a qualitative analysis
Structured mathematical models are required to describe the
MEOR processes in a better way. In order to develop a proper field
strategy, formulation of an efficient reservoir simulator capable of
predicting bacterial growth and transport through porous net-
work and the in situ production and action of the metabolites
called MEOR agents is of paramount importance. Several studies
have been reported on mathematical modeling of biomass
growth, selective plugging and bacterial transport in porous
media [79–83]. The simplified models were developed based on
fundamental conservation laws along with growth, retention
kinetics of biomass and biomass concentration in aqueous and oil
 ARTICLE IN PRESS
720 R. Sen / Progress in Energy and Combustion Science 34 (2008) 714–724
phases in order to predict porosity reduction as a function of
distance and time or based on a filtration model to express
bacterial transport as a function of pore entrance size and also to
relate permeability with the rate of bacteria penetration by
applying Darcy’s law [80–82]. From an order of magnitude
analysis, it is known that only biosurfactant flooding and selective
plugging mechanisms have the potential to recover more oil in
MEOR process. This is the major reason behind the formulation of
mathematical models of MEOR process based on these two
mechanisms. The basic equations governing the transport of oil,
water, bacteria, nutrient and metabolites in MEOR are based on
component mass balance and overall mass conservation.
The biomass-plug propagation and channel breakthrough
using Bingham yield stress of biofilm were described well by a
biofilm removal model, which represents the stability of biofilm
against shear stress. The simulation results showed that the
biofilm models based on Bingham yield stress predicted the
biomass accumulation and channel breakthrough reasonably well
[81]. A three-phase, multiple-species, one-dimension model was
developed to simulate microbial growth and transport, and
metabolism processes involved in MEOR and to predict perme-
ability modification that results from these microbial activities in
porous media. Convection dispersion equations and microbial
kinetics were incorporated in the model system to characterize
and quantify biomass production, product formation, and nutrient
utilization in the MEOR process [82]. The model was validated by
applying to static (sand packs) and core-flooding (sandstone
cores) experiments to describe microbial movement, metabolite
production, and nutrient consumption during growth and meta-
bolism and also to estimate permeability reduction. This could be
extended to provide numerical predictions for the purposes of
design and evaluation of MEOR field projects [82]. Some of the
relevant scaling up criteria and numerical simulation results were
discussed in order to explain the difficulties of scaling up
laboratory results before planning a field application [83]. The
experimental conditions of the MEOR technique applied for
Garzan oil (261 API; southeast Turkey) were utilized in a
mathematical model that describes the transport of bacteria and
its nutrients by convective and dispersive forces, including
bacterial decay and growth [84]. A recent study that generated
essential data for modeling in situ MEOR processes included
specific growth rates, carbon balances, biosurfactant production
rates and biosurfactant yields [95]. But a more comprehensive
approach based on artificial neural network modeling would be
more useful to develop suitable models for describing the in situ
MEOR processes.
6. Field trials
Microbial enhanced oil recovery technologies have progressed
from laboratory-based studies in the early 1980s to field
applications in the 1990s. The oil reservoirs worldwide represent
very complicated biological systems for which laboratory simula-
tions of microbial activities become very challenging. The
microbial consortia that are introduced into an oil field would
have to compete with the indigenous microflora. While it may be
possible to show beneficial effects in laboratory conditions with
tailored microbial cultures, in the reservoir these are likely to be
out-competed by the better adapted indigenous species. In most
of the cases, success or failure of field applications of microbial
technology is often hindered by the absence of specific and
quantitative understanding of microbial activity. A reservoir
engineering perspective, focusing on issues such as scale-up of
laboratory results, process design, and field implementation and
operation provides a consistent framework for comparing MEOR
with other EOR processes [87]. This justifies the rationale behind
the pilot scale field trials that are undertaken after successful
development of MEOR processes through R&D in laboratory
settings. Field trials to determine and document the effectiveness
of microbial processes and to assess the validity of laboratory
studies and models have been conducted. The application of
MEOR in these trials has resulted in a substantial and sustained
increase in production compared to control operations on the
same reservoir [88]. A pictorial presentation of the state of the oil
droplets in a porous sandstone rock before and after the MEOR
process is illustrated in Fig. 5.
In the last decade of the last millennium, a number of MEOR
field projects have been conducted in various parts of the world
with varying degrees of success. The field assessment of a MEOR
technology applied in the Alton field in Australia showed an
approximate 40% increase in net oil production, 12 months after
treatment, which sustained for considerable period of time [88].
Microbial enhanced water-flooding technology has also been
shown to be an economically feasible technology in the United
States [16]. Deliberations on information and papers presented by
companies and researchers all over the world on field applications
using microbes and also the details of MEOR projects have been in
public domain [11,16]. A compilation of very relevant information
and data of field trials conducted in the USA and Romania is
presented in Table 6.
An analysis of the data from a data base of information
collected from 322 projects employing the same MEOR methods
resulted in the evaluation of the technical effectiveness and
process economics of this particular technology and provides a
source of information useful for predicting treatment response in
any given reservoir [89]. The application of MEOR in these trials
has resulted in a substantial and sustained increase in production
compared to control operations on the same reservoir. Obviously,
there are differences in the extent of improvement in oil recovery,
which is influenced by various factors including individual
reservoir characteristics like lithology, nature of the sands,
porosity, permeability, reservoir temperature, crude oil gravity
and the drive types. In addition to the bottom-hole conditions and
the nature of the formations, bacterial composition and concen-
tration of the inocula, their adaptation time, optimum slug size
and the application time constitute the major governing factors in
MEOR. Efforts to explain these differences are severely limited by
the lack of quantitative measures of microbial performance in
terms of the reaction rates, stoichiometry, required product
concentrations, etc. It is possible, however, to demonstrate
quantitative relationships between microbial performance, reser-
voir characteristics and operating conditions [87]. An unfortunate
consequence is that the relevance of microbial performance to
reservoir engineering design is often obscure. Conversely, ex-
plaining success or failure of field applications of microbial
technology is often hindered by the absence of specific and
quantitative understanding of microbial activity.
In an MEOR field study in the Southeast Vassar Vertz Sand Unit
salt-containing reservoir in Oklahoma, nutrient injection stimu-
lated growth of the microbial populations, thereby decreasing the
effective permeability by 33% [90]. A biosurfactant flooding
process using a very low concentration (35–41 ppm) of biosurfac-
tant produced by Bacillus mojavensis strain JF-2 has been recently
reported to be very effective in recovering about 35–45% residual
oil from Berea sandstone cores [91]. Literature reports discussing
in situ applications of MEOR in field trials with analysis of the
results are available [17,18,89,92–95,98]. Very recently, a field
study involving in situ production and application of biosurfac-
tants by a consortium of Bacillus strains demonstrated that
approximately nine times the minimum concentration of biosur-
factant required to mobilize oil was produced in situ and resulted
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R. Sen / Progress in Energy and Combustion Science 34 (2008) 714–724 721

Fig. 5. Before MEOR process: oil is trapped in porous sandstone. Water flows without dislodging oil droplets from tiny pore spaces in the rock. After MEOR process:
microbes surround the oil droplets, causing them to dislodge from the microscopic pore spaces between the tiny sandstone and carbonate rock particles in the reservoir for
enhanced recovery (adapted with permission from http://www.titanoilrecovery.com/titanprocess1.html).

Table 6
Details of some field trials showing the efficacy of MEOR methods (adapted from Bass and Lappin-Scott [11])

Field test details Phoenix Pilot Project, NIPER SE Vasser Vez Sand Pilot Project, OU Institute of Biology, Romanian Academy

Year of initiation 1990 1991 1990

Oil field Chelsea-Alluwe SE Vasser Vertz sand unit Bragadiru
Formation Bartlesville sandstone Vertz sandstone Information not available
Depth (m) 122 550 780
Permeability (md) 16 60–181 150–300
Salinity (%) 2.9 11–19 0.06–0.3
Oil viscosity (cp) 6 2.9 9
Injection wells 19 5 treated 1 treated
Production wells 47 19 Information not available
Nutrients used Cane molasses Molasses, NH4NO3 Molasses
Microorganism(s) Bacillus, Clostridium Indigenous microflora Bacillus, Clostridium, Arthrobacter, Pseudomonas, Micrococcus
MEOR-Agents Biosurfactans, acids and gases Biomass and gases Biosurfactants, biopolymers, acids, gases and solvents
Water flood Yes Yes Yes
Test length 1.5 Years Approximately 9 months 4 Years
Pre-MEOR production 1 B/D per well No oil production before injection 1.5 B/D per well
Post-MEOR production 1.2 B/D per well 83 barrels produced (January 92–June 92) 7.4 B/D per well
Comments 20% increment in oil production Decreased permeability Cyclic microbial recovery, well-bore clean up

NIPER: National Institute of Petroleum and Energy Research, USA; OU: Oklahoma University.

in the recovery of substantial amount of oil entrapped in the
limestone reservoir of the Bebee field, Pontotoc City, Oklahoma,
USA [95].
The technical and economical feasibility studies of MEOR were
carried out in mature water-floods in continuation of a pilot study
of controlled microbial colonization in producer wells in La
Ventana oil field of Argentina [96]. The MEOR pilot projects
carried out in Providencia and Lobitos oil fields in Peru resulted in
36.5% and 46.5% increments in oil production, respectively, and
were considered profitable and economically feasible of further
expansion [97]. A new MEOR technology, termed as microbial
permeability profile modification (MPPM) technology, has been
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722 R. Sen / Progress in Energy and Combustion Science 34 (2008) 714–724
reported to improve oil recovery by adding nitrogenous and
phosphorus-containing nutrients to the injection water of a
conventional water-flooding operation [99]. This MPPM technol-
ogy has been claimed to have extended the economic life of the
field called North Blowhorn Creek Oil Unit (NBCU) located in
Alabama by 60–137 months, with an expected recovery of
(4–6)  105 bbl of additional oil. Another microbial profile mod-
ification method employing a spore-bearing halotolerant meso-
philic bacterium has been reported. This bacterium produces
spores that can propagate easily in Berea cores with permeability
4500 md and can reduce the rock permeability by producing
biofilm on germination under suitable nutritional and environ-
mental conditions [100]. Economically attractive alternative EOR
technologies including microbial EOR, have been discussed and
numerical simulation results have been presented to demonstrate
the difficulties in scaling up and translating the laboratory results
to real-time oil field settings [101]. Comprehensive R&D studies
and annual reports on the development of microbial strains with
improved transport and biosurfactant activity for EOR and on the
biosurfactant-mediated oil recovery in model porous systems
with its computer-aided simulations have tremendously contrib-
uted to the design and development of effective MEOR strategies
and at the same time have enormously enriched the scientific
literature on MEOR [102–104].
Applications of MEOR processes in the oil fields in Asian
countries, for example in Chinese, Malaysian and Indian oil fields,
have been reported [92,105–110], though published literature
does not have field trial data in an organized format as shown in
Table 6. In Malaysia, the estimated oil-in-place from the 47
producing fields stands at about 20.1 BSTB (billion stock tank
barrels), with a cumulative production of 4.9 BSTB till 2003 [105].
However, there were no published reports on full-fledged field
application of MEOR in Malaysia, with the exception of an MEOR
stimulation project in Bokor field [105]. There is one structured
report on MEOR field trial in the Fuyu oil field, Jilin province,
China. The trial was carried out in 2001 in the sandstone
formation with an average permeability of 180 md at a depth of
300–500 m and an oil viscosity of 6–8 cp. The recovery of oil could
be doubled after 6-month MEOR treatment from 14 production
wells by injecting a bacterial consortium containing Enterobacter
cloacae and Bacillus licheniformis as major bacterial strains and a
nutrient package mainly comprising of molasses (10%) and
cellulase enzyme (0.01%) [106]. Microbiological and chemical
characteristics of the production fluids of the high-temperature
Liaohe oil field in China were examined to investigate the
potential for MEOR [107,108]. It was concluded that the Liaohe
oil field contained a diverse thermophilic microbial community
having a high biotechnological potential for petroleum recovery
[107]. Recently, a study to assess the potential applicability of
MEOR techniques in the Kongdian reservoirs of the Dagang oilfield
in China showed that the oil field was inhabited mostly by
anaerobic thermophilic fermentative, sulfate-reducing and
methanogenic bacteria and hence, had great potential for
application of MEOR processes by activating the stratal microflora
[109]. In an exploratory study with the formation water from
Daqing oil field, China indicated that the oil field was found to be
inhabited by aerobic bacteria like Bacillus, Pseudomonas, Rhodo-
coccus, Dietzia, Clavibacter, etc., which were capable of producing
effective oil-releasing agents including biosurfactants [110]. A
laboratory study involving growth of some selected strains in the
crude oil samples and consequent reduction of oil viscosity
indicated that the results could be translated into an effective
strategy for recovering residual oil from Daqing oil field, the
biggest oil field in China [111]. The MEOR squeeze treatment,
which was carried out for about 35–40 days in 1997, was found to
have increased production performance and decreased water cut
in 17 wells out of 25 wells treated. Another laboratory study using
a model reservoir simulating the conditions of Daqing oil field and
a pilot study in some wells of the same oil field employed the
metabolic products (PIMP) from a strain of Pseudomonas aerugi-
nosa, which showed significant increment in crude oil recovery
and prolongation of the cycle of oil well washing to increase total
oil production [112].
In India, the Oil and Natural Gas Corporation (ONGC) Limited,
in collaboration with The Energy and Resources Institute (TERI,
New Delhi) and the Institute of Reservoir Studies (IRS), Ahmeda-
bad, conducted some field trials by employing a Huff and Puff
process and using an indigenously developed MEOR technology
based on a consortium of anaerobic extremophiles isolated from
the candidate reservoirs. A three-fold increase in oil production
and a significant reduction in water cut were achieved by applying
this technology in 9 wells out of 12 wells treated in 4 oil fields,
mostly in the state of Gujarat [113]. Recently, a US patent has been
granted on the same process and microbial consortium, which
was developed in TERI in collaboration with IRS and was evaluated
in field trials by ONGC [114].
7. Conclusions
MEOR represents a truly eco-friendly petroleum recovery
process employing biotechnological resources and techniques
that can be used to replace and augment the traditional EOR
processes and flooding chemicals. Many countries have envisaged
that one-third of their oil recovery programs will utilize MEOR
techniques by the year 2010. Though it has been repeatedly
encountered that the benefits of MEOR projects applied to one
well have positively affected the recovery performance of
neighboring wells, thus improving the process efficiency; better
understanding of the reservoir characteristics, biochemical and
physiological characteristics of microbial consortia, controlling
mechanisms and process economics are essential before MEOR
becomes a viable process for general field applications. Most of
the MEOR work leading to field trials has been completed in the
last two decades and now the technology has advanced from a
laboratory-based evaluation of microbial processes, to field
applications internationally. Though research initiatives and
endeavors in MEOR have progressed rapidly, still some questions
remain unanswered. In spite of microbial processes holding great
promise and prospect for EOR, the reasons that retard the
implementation of MEOR include inconsistency in in situ
performance, low ultimate oil recovery factor, uncertainty about
meeting the engineering design criteria by microbial processes
and a general apprehension about processes involving live
bacteria. It is now known that manipulation of microbial consortia
activity is a potentially powerful process that can profoundly and
beneficially affect the microenvironment of a reservoir to mobilize
more oil, increase production rates and technical efficacy of the
recovery systems. In situ application of a judicious consortium of
aerobic and anaerobic bacteria, acclimatized in simulated oil
reservoir conditions, coupled with the bioaugmentation and
stimulation of indigenous microflora have been successful in
enhancing both the oil productivity and ultimate oil recovery
factor. However, a better understanding of the MEOR processes
and mechanisms from an engineering standpoint based on
economics, applicability and performance is the key to further
improve the process efficiency for writing more success stories.
Advanced MEOR technologies based on the use of genetically
engineered organisms and some specific enzymes may prove to be
very effective and successful in the future. Nevertheless, an urge
and urgency in developing novel MEOR technologies in the wake
of an impending energy crisis seem to be imminent, mainly on
 ARTICLE IN PRESS
R. Sen / Progress in Energy and Combustion Science 34 (2008) 714–724
account of their perceived potential to offer a cost-effective
solution to the conventional EOR strategies.
Acknowledgments
I thankfully acknowledge the financial support that I have
received from the DBT (Grant no.: BT/PR-6827/AAQ/03/263/2005),
Government of India and my institute, IIT, Kharagpur (ISIRD
Grant). I am also thankful to my parents, my wife, Anamika, and
our little son, Yuvraaj Sen, for their love, support, forbearance and
encouragement to complete this comprehensive review. I am
grateful to Titan Oil /http://www.titanoilrecovery.com/S for
allowing me to use some of their pictorials.
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