Effect of Bacterial Polysaccharide

Production on Formation Damage

R.E. Lappan and H.S. Fogler, SPE, U. of Michigan

Summary. In-situ growth of cellular material is known to cause formation damage. Bacterial reproduction and polysaccharide pro-
duction are the key factors that segregate bacterial formation damage from fines and particulate damage. Carefully controlled experi-
ments conducted on both high- and low-permeability ceramic cores showed that bacteria can plug the pore space and damage the cores.
However, further experimentation demonstrated that polysaccharide production is largely responsible for this damage. This conclusion
is based on a comparison of two experimental systems: core plugging from bacterial replication and polymeric production and plugging
of the porous medium caused solely by cell division with no polysaccharide production. In light of these results, the interpretation of
reservoir plugging resulting from the presence of bacteria requires further scrutiny.

Introduction
Bacterial growth and transport in reservoirs can greatly influence
EOR.I,2 Bacteria can enhance recovery by producing gases (C0 2
and H 2) that increase the well's internal pressure and decrease the
viscosity of the crude oil (below bubblepoint). 3 In addition, in-situ
cellular biosurfactant production has been shown to be a viable tech-
nique to enhance oil yields by decreasing the interfacial tension
between the oil and the water, thus allowing better crude emulsifi-
cation. 3 ,4 These two phenomena are but a few of the possible bac-
terialhydrocarbon interactions that can influence oil recovery. These
phenomena have been given the collective name of microbial EOR
(MEOR), a tertiary oil recovery technique.
The plugging of a water-swept zone by the injection and cultiva-
tion of bacteria, in situ, has also been proposed 5 as an economi-
cal technique for flow diversion. This technique would be initiated
with the injection of bacteria, suspended in a nutrient-rich medi-
um, into the reservoir. Next, the well would be shut in to provide
sufficient time for microbial growth, hence plugging the high-
permeability thief zones. Then water injection would be resumed
and the plugged zone would divert the injection water into unswept
zones.
Rationale. Before any MEOR technique, such as in-situ surfac-
tant and gas production or flow diversion, can be realized, a basic
understanding of how bacteria are transported through porous me-
dia and how their retention affects the permeability of the media
is needed. Bacterial transport through and retention by porous me-
dia differ from particle transport because cells increase in number
and can produce polysaccharides, which affect their ability to ad-
here to surfaces. 6-9
Bacterial adhesion experiments with inert nonporous surfaces
demonstrated the importance of polysaccharide production.~-9
These studies, however, considered only the effect of polysaccha-
rides on cell adhesion to inert nonporous surfaces and did not con-
sider the effects of in-situ growth. Consequently, there is a lack
of information on the effects of polysaccharides on cell transport
and retention in porous media. This study, which is part of a series
of experiments demonstrating bacterial cell transport and retention
in porous media, was conducted to determine the effects of poly-
saccharide production on reservoir plugging.
Background
Bacterial Adhesion. It has been proposed that bacterial adhesion
is a two-step process. 1O,1l The first step is the reversible adsorp-
tion of bacteria to the surface. This initial contact of the cell with
the surface is dependent on the colloidal, hydrodynamic, and iner-
tial forces. 12 Once the cell is adsorbed on the surface, the second
step begins with the polysaccharide production, leading to the ir-
reversible binding of the cell to the surface. Fletcher et at.7 ob-
served the irreversible binding phenomenon in experiments on the
adhesion of marine bacteria to filters. They discovered that it is
not the cell's primary acidic polysaccharide coating that irreversi-
Copyright 1992 Society of Petroleum Engineers
SPE Production Engineering, May 1992
bly binds the cell to the surface, but the production of a secondary
acidic polysaccharide that firmly attaches the cell. The primary poly-
saccharide helps in the initial adhesion of the cell by mediating a
favorable contact between the cell and the surface. The secondary
polysaccharide, a more fibrous reticular (net-like/tangled) substance,
interconnects between and around adjacent bacteria or between bac-
teria and the surface to attach the cell irreversibly. Fowler et at.
demonstrated this phenomenon by subjecting cells (attached to glass
and stainless steel surfaces) to various shear stresses in a radial flow
chamber. 8 It was found that the shear stress required to suspend
the sessile (attached) bacteria increases with the contact time given
to the cells for the adhesion.
Bacterial Injection and In-Situ Growth. Earlier microbial trans-
port experiments used dead bacteria for core-injection studies. Typi.-
cally, common bacteria, such as Bacillus subtilis, were grown, killed
by sterilization, and then injected into a Berea sandstone core. 13
Results from the injection of dead bacteria into the sandstone core
indicated that bacteria would enter the porous medium and be trans-
ported until they were sieved by a pore of smaller dimensions or
captured according to deep-bed filtration mechanisms. 14 Sieving
is the dominant capture mechanism that occurs within the inlet sec-
tion of the core during the initial stages of bacterial injection. Dur-
ing injection of the dead bacteria, blocking the larger pores takes
longer than blocking the smaller pores because the bacteria must
first agglomerate. 13 This plugging sequence produces the reverse-
S-shaped curve when the permeability ratio, k/ko (where ko = initial
permeability of the core before cell injection), is plotted as a func-
tion of time (or PV's injected) on the abscissa (Fig. 1). In addi-
tion, the injection of dead bacteria into Berea sandstone exhibited
the development of an external filter cake at the injection face of
the porous medium. IS
The injection of live bacteria into core samples also produces the
typical reverse-S-shaped permeability reduction curve. 16 Closer
examination of these injected core samples with a scanning elec-
tron microscope (SEM) have shown that the pores were plugged
by bacteria coated with exocellular polysaccharide. Cores injected
with dead cells, when examined by SEM techniques, did not dem-
onstrate this phenomenon. In addition to these observations, the
development of an external filter cake was noted during the injec-
tion of live cells. 16
Once the bacteria enter the reservoir, they may grow in situ. This
growth is dependent on the availability of nutrients within the reser-
voir. In secondary oil recovery, where large volumes of water are
injected into the reservoir, accelerated indigenous bacterial growth
can be supported by the nutrients in the injection water, thus caus-
ing near-wellbore plugging. Bright-field microscopic examination
of test cores flooded with samples of reservoir injection water (water
known to contain bacteria and growth nutrients) indicated that the
pores at the injection face of the cores become plugged owing to
the capture, followed by the growth, of planktonic cells in the in-
jection water. 17 Accompanying the cells were large amounts of
cellular polysaccharide. For MEOR, nutrients are injected into the
formation, typically preceding bacterial cell injection, to induce in-
167
 0.10 1.0"",
( max. _erlllol..)

c
.2
0.08
I
I, \ Low
U 0.08
~ r \ "'rmaablllly
eore
I!
, ..
High
I \ .........bUIly
~ "" 0.04 eore
0.02 ...
0.00
10 20
-------
30 40 50 60
Pore size (J.lm)
o
Time or pore volume
Fig. 2-Pore-slze distribution for low- and high-permeability
cores (distributions based on log-normal and normal proba-
Fig. 1-Reverse-S-shaped permeability reduction curve ob- bility cumulative curve fits of mercury poroslmetry data, re-
served during live and dead bacterial InJection. spectively).

situ cell growth and bioproduct production. Cellular growth means physical characteristics of Leuconostoc mesenteroides. In short, one
increased biomass production and thus reduced reservoir injectivi- may consider these bacteria to be self-replicating, polymer-
ty. Hence, the development of an injection strategy that allows cel- producing colloidal particles, typically I I'm in size.
lular growth within a formation and minimizes formation damage
may make MEOR processes technically feasible. Media Selection. The core-plugging experiments started with the
The use of bacteria for the plugging of water-swept zones by in- injection of the model bacteria into a well-characterized porous
situ growth has one foreseeable drawback, namely the problem of medium. Consolidated porous ceramic cores were selected as an
cell growth in the near-wellbore region. It is desired that the bac- ideal medium for this study because they exclude such artifacts as
teria grow exclusively in the water-swept zone and not in the near- fines migration, have a uniform surface chemistry composition, and
wellbore region. One proposed technique is the limitation of the have a specific and uniform pore-size distribution. Table 2 lists
cell's ability to produce polysaccharides. IS Cells can be manipu- the chemical and physical characteristics of the ceramic cores.
lated so that their ability to produce polysaccharide could be in- Uniformity in chemical composition, and hence surface chemis-
duced or hampered by controlling the type and amount of nutrient try, is important in the initial phase of the experiment. Deep-bed
feed supplied to them. Hence, a potential cell injection procedure filtration theory predicts that the capture of submicron particles (0.1
for treating a water-swept zone would be to inject the bacteria un- to 1 I'm) is primarily the result of the colloidal forces that exist
der restricted polysaccharide-producing conditions and then to in- between a particle (the bacterial cell) and the surface of the porous
troduce a feed that would induce cellular polysaccharide production media; 20 Hence, for the sake of comparison between polymer-
in the high-permeability zone. Although the idea of controlling bac- producing and non-polymer-producing experiments, the injection
terial growth in situ holds promise, previous experiments did not and eventual capture of the bacteria must be consistent between core
differentiate between the plugging of the core samples by polysac- samples for proper interpretation of results.
charide production or by cell growth. Hence, there is a need to A specific pore-size distribution was selected as an important
determine the relative effects of both cellular growth and polysac- criterion because such effects as bacteria straining on the initial in-
charide production on reservoir plugging. jection of bacteria should be minimal. Bacteria straining is a unique
retention phenomenon, and its occurrence would make it difficult
Methodology to interpret the results from experiments designed to demonstrate
To determine whether cellular polysaccharide production or cell polysaccharide plugging. Also, the initial bacteria injection must
production is the dominant plugging mechanism, a series of ex- not cause serious damage to the medium. Therefore, the porous
periments using a model bacteria for injection and in-situ growth medium must have a mean pore diameter that is greater than the
in a well-characterized porous medium was conducted. size of the bacteria. For this study, ceramic cores with two pore-
size distributions were selected. Fig. 2 presents the results obtained
Bacterial Selection. For this study, the genus Leuconostoc mesen- from mercury porosimetry determination of the pore-size distribu-
teroides was selected as the model bacteria because it has the unique
ability to produce dextran, a water-insoluble polysaccharide, when TABLE 2-PHYSICAL AND CHEMICAL CHARACTERISTICS
it is fed sucrose. When this bacterium is limited to a glucose/fructose OF THE CERAMIC CORE
feed, however, no dextran is produced, so cellular polysaccharide
production can be controlled. This controlled production enables Percentage
the formulation of experiments that demonstrate the effects of in- Mineral (wt%)
situ polysaccharide production on the permeability of porous me- AI 2 0 3 50
dia by conducting parallel plugging experiments where polymer pro- Si0 2 37
duction is promoted or repressed. Table 1 lists some of the relevant CaO 7.7
MgO 3.8
TI0 2, Fe203' K 20. Na20 <1% each
TABLE 1-SUMMARY OF THE PHYSIOLOGICAL
CHARACTERISTICS OF -LEUCONOSTOC MESENTEROIDES 19 High Low
Permeability Permeability
Mesophilic Porosity 0.555 0.536
Cocci 0.5 to 1.2 I'm in size Permeability. darcies 14.7 0.098
Aerotolerant Mean pore size, I'm 33.2* 3.27**
Anaerobic Standard deviation 12.1 0.94
Agglomerate Group(s) of 1, 2. or 4
Dextran production When fed sucrose 'Based on normal distribution.
No dextran production When fed fructose or glucose "Based on log·normal distribution.

168 SPE Production Engineering, May 1992

 TABLE 3-GROWTH MEDIUM
to p ........ glUgH
Water, mL 900 and data Icqutaltlon
Tryptone, 9 20
Yeast extract, 9 5
NaCI, 9 4
Sodium acetate, 9 1.5
Ascorbic acid, 9 0.5
Trace elements Ca, Mn, Fe, Mg
Carbohydrate solution·
Distilled water, mL 100
Sucrose, 9 15.0
Or a combination of
fructose, 9 7.9
and glucose, 9 7.9
Yeast extract (and tryptone)are
supplied to the cells as the sources of amino acids,
purines, pyrimidines. and vitamins, which are needed by the cells for growth
because Leuconostoc cells cannot synthesize these compoun~s for themselves.
Fig. 3-Hassler™ cell used for bacterial transport experi-
'The carbohydrate solution is mixed with the other ingredients after autoclaving. ments.

tion, while Table 2 presents the calculated means and standard devi-
ations. Note that both cores have a mean pore size that is larger
than the size of a Leuconostoc mesenteroides cell. For the high-
permeability core, it is desirable for the bacteria to be distributed
evenly throughout the core. The low-permeability core was select-
ed to emulate a tYPical reservoir permeability that would allow cell
transport 21 rather than having an even distribution of bacteria
axially.
Experimental Procedures. The plugging experiments consisted of
two phases: the initial injection of bacteria into the porous medium
followed by the injection of a nutrient feed into the medium to pro-
mote cell and (depending on the type of saccharide added) poly-
mer production. These experiments were conducted on both the
high- and low-permeability cores.
The inoculum cells were grown in a glucose/fructose feed solu-
tion as a batch culture for an 18- to 24-hour period and reached
a cell density of about 10 9 cells/mL, as measured by plate counts.
Table 3 lists the constituents supplied in the glucose/fructose medi-
um. This culture, cell and spent-medium solution, was then inject-
ed into the core samples (2.54 em diameter by 5 cm in length) at
a constant flow rate of 5 mLimin, which translates to an injection
superficial velocity of 0.987 cm/min. After the inoculation of the
core, a glucose/fructose feed was injected into the core at a rate
of 0.1 mLimin (1.18 cm/h). The pressure drop across the core was
then monitored for the remainder of the experiment. This experi-
ment was then repeated, except sucrose feed was used instead of
the glucose/fructose feed (see Table 3).
Fig. 3 shows the apparatus used in this study. This equipment
includes the self-contained piston container, nutrient prefIlter, and
pressure transducer fIlters. These pressure transducer fIlters main-
tain the system in an aseptic condition and were found to have
minimal effects on the response rate of the pressure transducers.
Initially, the apparatus was sterilized by ethanol (200 proof) rins-
ing or autoclaving.
Results
Figs. 4 and 5 show the results of the plugging of the porous media
for both high- and low-permeability core samples. The permeabil-
ity reduction calculated from the data is plotted as a function of
time. The results from the high-permeability core experiments have
been replicated on three separate occasions. As Fig. 4 illustrates,
the injection of the bacteria (inoculum) into the high-permeability
cores did not affect the damage or plug the porous media. The lower-
permeability coreS, however, did show a two-thirds reduction in
permeability when the inoculum was injected. Fig. 5 shows the
reduction in permeability of the low-permeability cores at the start
of the feeding when k/k o '"" 0.3 to 0.4.
Figs. 4 and 5 also illustrate the effects of injecting the cores with
the feed solutions. The permeabilities of the sucrose-fed cores were
reduced more drastically than those of cores fed glucose/fructose.
These results show that polysaccharide production causes increased
core plugging, if cell growth rates for sucrose-fed cells are of the
same magnitude as the rates when the cells are supplied glu-
cose/fructose feed. A kinetic study proved this to be true. That is,
the doubling times for the bacteria under the different feeds are
approximately equal. Virtually equal growth rates were expected
because the nutrient feeds were stoichiometrically equal; i.e., for
every mole of sucrose in the polysaccharide-promoting feed, there
is a mole of glucose and a mole of fructose in the non-polymer-
promoting feed. (Note that sucrose is a disaccharide composed of
two monosaccharides, glucose and fructose, joined by a glycosid-
ic bond.)

0.1

0 0.1 • Glucose-Fructose 0
~

~ • Sucrose 0.01
i2
0.01
0.001

••
0.001 Glucose-Fructose
Sucrose
0.0001 0.0001
0 10 20 30 40 50 60 70 80 0 5 10 15 20 25 30 35 40

Time (hr) Time (hr)

Fig. 4-Permeablllty reduction caused by dextran synthesis Fig. 5-Permeablllty reduction caused by dextran synthesis
In high-permeability (14.7-darcy) core. in low-permeability (98-md) core.

SPE Production Engineering, May 1992 169


500 Glucose-Fructose Sucrose
Feed Feed
400
:i
!3OO - - Inlet Pre.....
_ .. ,30m
.~200
! 100
.... Pre..... 3.8 om
~
0
0 20 40 60 60 100 120 140
Time (hr)
Fig. 6-lnlet and Interstitial pressure results during glu-
cose/fructose feeding followed by sucrose feeding (at a flow
rate of 0.1 mUmln).
It may also be debated that the cells injected into the high-
permeability cores and then fed glucose/fructose did not plug the
porous media because they were washed out of the core. An addi-
tional experiment was then conducted to demonstrate that the cells
were present in the cores and could cause damage when provided
a sucrose feed. This core-plugging experiment was conducted with
a high-permeability (5-darcy) core injected with an inoculum and
then fed a glucose/fructose solution under the same conditions stated
above. Then after 56 hours, the feed was switched from a glu-
cose/fructose feed to a sucrose solution, thus producing the results
given in Fig. 6. In this plot the inlet, 1.3-cm, and 3.8-cm axial
pressures (distances measured from inlet) are shown as a function
of time instead of the overall permeability reduction of the core.
The resulting pressure increases provided by the graph clearly dem-
onstrate that cells are retained by the porous media and that they
can plug the media when provided sucrose. In addition, Fig. 6 il-
lustrates an increase in the 13-cm pressure tap reading after 110
hours of feeding (54 hours of sucrose injection). This increase in
pressure means that the cells are not just plugging the inlet face of the
core but are penetrating well within the core and causing damage.
After completion of these experiments, the cores were halved
axially and dyed with crystal violet. Crystal violet is a stain that
specifically dyes gram-positive bacteria. The intensity of the color
produced from the staining is proportional to the cell concentra-
tion in the local region. Hence, this staining procedure gives the
concentration gradient of the bacteria throughout the length of the
core. Fig. 7 shows the relative axial cell concentration measured
by crystal violet stain for the high-permeability core. The relative
cell concentration is based on the concentration of cells (or intensi-
ty of violet color) divided by the concentration of cells at the inlet
of the sucrose-fed core. As depicted in Fig. 7, the cells in the glu-
cose/fructose-fed cores are relatively evenly distributed through-
out the core compared with the sucrose-fed core, which exhibited
a large cell concentration gradient. A third core was also injected
with cells but was not supplied a feed. The core was cut and stained
to determine the cell deposition pattern within the high-permeability
cores. The staining of this unfed core demonstrated a linearly dis-
tributed low concentration of cells through the core, indicating a
lack of cell-transport hindrance by the high-permeability porous
medium. The results obtained from these three core stainings sug-
gest that the polysaccharide-producing cells grew at the injection
face, where nutrient availability is high, and were retained there.
The non-polysaccharide-producing cells were not retained as easi-
ly by the porous medium' and thus were able to be transported within
the core, reducing the magnitude of the gradient.
The staining of the low-permeability cores did not show such a
drastic cell gradient. This may be explained by the fact that mech-
anisms other than cell retention caused by polysaccharide produc-
tion are significant in this core.
Discussion
The core-plugging results have significant ramifications regarding
the implementation of any MEOR processes. From these core-
170
1.0
• Sucrose
.. Glucose-Fructose
0.9
_t:
-0
c5i 0.8
QI-=
>t:
i~
- t : 0.7
Ql
Et:O
O
0.6
0.5
0.2 0.4 0.6 0.8
0
Dimensionless Length
Fig. 7-Axlal cell concentration (relative to the Inlet cell con-
centration of the polysaccharide-plugged core).
plugging results, it is now known that the use of any MEOR tech-
nique in a low-permeability reservoir will always exhibit forma-
tion damage because of the combined effect of cell growth and
polymer production. Hence, the development of an MEOR tech-
nique for a low-permeability reservoir must promote the produc-
tion of biogases and biosurfactant, which are the desired bacterial
products, and retard the generation of large quantities of cells and
polysaccharides. For high-permeability reservoirs, the restrictions
are a little more lenient. Cells would be allowed to replicate dur-
ing synthesis of the biogases and biosurfactant, but again polysac-
charide production would have to be eliminated or minimized to
prevent damage to the reservoirs.
In addition, the staining results also have dramatic consequences
regarding the application of any MEOR process. The adaptation
of an MEOR process, which would require the maintenance of cells
at the oil/water interface to maximize oil recovery, would have to
control or eliminate the cell's ability to produce polysaccharides
because the biopolymer radically hinders the cell's transportabili-
ty. Whether this control can be accomplished by preventing the
generation of polysaccharides or the degradation of polysaccharide
depends on the bacterial strain selected for MEOR.
These core plugging results can also be used to further our under-
standing of formation damage caused by biomass production (cell
and polysaccharide production). As Geesey et al. 17 stated, flood-
ing water will typically contain nutrients that can support both cell
growth and polysaccharide production by cells either suspended in
the injection water or indigenous to the reservoir. Hence, the re-
sults of plugging experiments suggest that nutrients leading to poly-
saccharide production should be eliminated or limited as a minimal
control of formation damage. In addition, for low-permeability
reservoirs, stringent water quality control would be needed to pre-
vent both polysaccharide production and cell growth.
Conclusions
The results of the cell-growth experiments show that cellular poly-
saccharide production is a significant factor in formation damage
by bacteria. Such damage was pronounced for the high-permeability
cores, where damage to the cores occurred only as a result of poly-
saccharide production. In contrast, the low-permeability cores dem-
onstrated damage from both polysaccharide and cell production.
Nonetheless, for the low-permeability cores, the permeability reduc-
tion with polysaccharide production was more than 10 times great-
er than that without polysaccharide production. The results from
the core staining also suggest that the transport of cells through
porous media is highly dependent on the cell's ability to produce
polysaccharides.
Acknowledgment
We gratefully acknowledge the Industrial AffIliates Program at the
U. of Michigan for providing financial support for this work. The
Industrial Affiliates Program consists of Chevron Oil Field Research
Co.; Conoco Inc., Halliburton Services, Japan Nat!. Oil Corp.,
SPE Production Engineering, May 1992
Marathon Oil Co., Mobil R&D Corp., Phillips Petroleum Co., Tex-
Authors
aco Inc., and Unocal Corp. In addition, author Lappan thanks his
colleagues at the U. of Michigan for their support. Raymond E. Lap-
pan Is a PhD-
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SPE Production Engineering, May 1992 171