SPEIDOE

SPEIDOE 24203
The Effect of In-Situ Pore Pressure on MEOR Processes
G.E. Jenneman and J.B. Clark,Phillips Petroleum Co.

Copyright 1592. Society of Petroleum Engineers Inc.

This paper was prepared for presentation at the SPElDOE Eighth Symposium on Enhanced Oil Recovery held in Tulsa. Oklahoma. April 22-24. 1992.

This paper was selected for presentation by an SPE Program Committee following review of information contained in an abstract submitted by the author@).Contents of the paper.
as presented, have not been reviewed by the Society of Petroleum Engineers and are subject to correction by the author@).The material, as presented, does not necessarily reflect
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ABSTRACT of hydrostatic pressure but found to be

absent under atmospheric conditions. The fact
Most laboratory work in MEOR, both in that some of the observed effects could be
screening and in development, is performed demonstrated by simply reducing the gas
under i n s i t u temperatures and in brines headspace suggests that gas solubility could
similar to those of the target reservoir. be an important factor. Gas formation in a
However, the effect of i n s i t u pore pressures core was estimated to account for as much as
in such work is normally ignored. Much of 50% of the permeability decrease observed in
our understanding of the effects of cores run under atmospheric pressure.
hydrostatic pressure on microorganisms comes Therefore the changes observed are of such
from studies of microbial communities at the magnitude as to alter the MEOR process in
ocean floor. Since nearly 90% of the ocean the reservoir from that indicated by
floor has pressures ranging from 10 MPa to laboratory studies not done under pressure.
over 100 MPa, there is a paucity of data
concerning effects on microorganisms at the Our results strongly suggest that all MEOR
lower pressures found in many oil reservoirs. studies pertaining to a known reservoir
should be evaluated under the i n s i t u
In our studies of the indigenous cells from conditions of the reservoir, including
injection brine at the North Burbank Unit, we pressure.
have found that pressures ranging from 0.1
MPa to 8.8 MPa can have a significant effect INTRODUCTION
OTI pH changes, substrate utilization, end
product formation andpermeability reduction. A number of mechanisms have been proposed for
In general, hydrogen ion concentrations and the recovery of residual oil through the
carbohydrate utilization rates increasedwith application of microbially enhanced oil
increasing pressures. Methanol was found to recovery (MEOR). Such mechanisms include: gas
be a significant end-product in the presence repressurization, production of
biosurfactants to reduce interfacial tension,
production of polymers to improve mobility
References and figures at end of paper. and increase sweep efficiency, production of
2 THE EFFECT OF IN SITU PORE PRESSURE ON MEOR PROCESSES
biogenic acids to dissolve carbonate rock,
etc. Without exception, the in s i t u
application of processes used to accomplish
these mechanisms requires the growth and/or
metabolism of microorganisms in the rock
matrix. Whether the microorganisms used to
carry out these functions are indigenous or
added exogenously, they must function
effectively at in s i t u conditions of
salinity, temperature, and pressure.
The importance of salinity and temperature on
microbial activity has been well documented;
however, the significance of pressure as a
metabolic effector is less understood and
generally believed not to be significant at
hydrostatic pressures under approximately 100
atrn (10.1 MPa). Consequently effects of
pressures below 100 atrn (10.1 MPa) are
assumed to be minimal or non-existent.ZoBell
and Hittle (1969) stated that the growth of
most species of marine, starch-hydrolyzing,
bacteria that they studied were retarded by
pressures exceeding 100 atrn (10.1 MPa). Only
a few reports ( Johnson and Lewin(1946),
Marquis and Matsumura(l978) and Heinritz et
a1 (1990)) have actually examined the effect
of pressures at or below 100 atrn (10.1 MPa)
on microbial growth. In these cases the
effect of pressure tended to slightly enhance
bacterial growth rate and in one case
enhanced growth yield above that reported at
1 atrn (100 kPa) . However the majority of
research involving life under pressures deals
with ocean floor communities in the proximity
of hydrothermal vents where 90% of the
pressures are over 100 atrn (10.1 MPa).
Likewise many oil reservoirs have pressures
in excess of 100 atrn (10.1MPa) but Jenneman
(1989) reported that 40 - 70% of the
reservoirs they surveyed for applicability to
MEOR in California, Oklahoma, and Kansas
exhibit pressures below 120 atrn (12.2 MPa).
Therefore a significant MEOR potential
resides in reservoirs with pressure below 100
atrn (10.1 MPa).
In addition to the paucity of pressure
studies involving microbial activities many
of the studies performed deal with effects on
pure cultures of microorganisms. It is well
accepted that oil field brines contain
complex communities of microorganisms that
interact both positively and negatively to
each other in response to nutrient input.
SPE 24203
Therefore the purpose of this paper was to
describe how low pressures (i.e. 0.1 - 10.1
MPa) , such as those found at the North
Burbank Unit (Shidler, OK) effect the
metabolism of natural assemblages of
microorganisms found in the injection brine.
The impact of these effects is discussed with
regard to current laboratory experimentation
to be used in implementation of MEOR field
tests.
MATERIALS AND METHODS
Brine Collection: Brine used in these
experiments was collected as effluent from
the bottom of a free water knockout tank
located at the tract 5 tank battery (North
Burbank Unit(NBU), Shidler, OK). This brine
is a composite of produced brine from a
number of area wells and is representative of
the brine being re-injected into nearby
injectors. Brine was collected in clean,
sterile, oxygen-free, glass bottles via
rubber or nylon tubing clamped to a 1/4 inch
(63.5 mm) nipple. The nipple was connected to
a 2.0 inch (50.8 mm) gate valve via a
bushing. When sampling, the valve was opened
allowing brine to purge the sampling line for
approximately one minute. The tubing was then
either placed inside a two-liter,wide-mouth,
screw cap bottle (Corning Glass) and filled
to the top or brine was introduced into a 32
ounce (946 cm3), glass, crown top bottle
(Texberry Container) via a syringe needle (21
gauge) attached to the end of the hose that
was pierced through a butyl rubber septum
lining the bottle cap. Another syringe needle
was pierced through the septum to serve as a
vent needle. Bottles were made oxygen-free by
pre-incubating inside an anaerobic glove box
(Coy Laboratory Products, Inc. , Ann Arbor,
MI) for at least 48 hours. Brine samples were
kept at ambient temperature and transported
back to the laboratory within three hours of
collection. A typical chemical analysis of
Tract 5 brine is presented in Table 1.
Density values for the brine of 1.08 g/cm3
(1080 kg/m3) and a charge balance of the ions
suggest that the actual total dissolved
solids are closer to 100 g/L since the
standard drying procedures used to dehydrate
brine containing high levels of divalent
metal cations are not usually effective.
SPE 24203 G. E. JENNEMAN AND J. B. CLARK
Description of barokams: Figure 1 is an
illustration of a barokam. A barokam is a
term used to describe a pressure vessel as
suggested by C. E. ZoBell ( Marquis, 1976).
A maximum of six barokams were run at any
given time. Barokams were made from one inch
(254 nun) NPT 316 stainless steel(SS) tees
connected on one end via 1/4 inch (63.5 mm)
SS tubing to a 100 cm3 SS accumulator (Welker
Engineering) containing a piston and on the
other end connected via 1/16 inch (15.9 mm)
SS tubing to a metering valve or a back-
pressure regulator. A pressure tolerant pH
probe (see pH measurement) was inserted into
the remaining opening of the tee.
All barokams were housed inside an oven for
temperature control and pressurized by means
of an hydraulic hand pump (Model 87-6-5,High
Pressure Equipment Co . , Erie, PA) connected
to the back side of the barokam accumulators
(Figure 2). All hydraulic lines were filled
with oxygen-free water housed inside an
anaerobic glove box. When sampling, the
contents of the barokams were extruded by
pressurizing the accumulator piston until the
system pressure exceeded the setting of the
back pressure regulator or until the system
pressure exceeded the resistance created by
the metering valve needed to hold the desired
test pressure.
Preparation and filling of barokams: All
parts of the barokam, except O-rings and
piston, were soaked in a 10% solution of HC1
and then rinsed with deionized water. The
parts of barokam were then sterilized by
autoclaving at 121 C for 15 minutes. The
barokams were then allowed to cool before
being placed in an anaerobic glove box. The
barokams were pre-incubated in the glove box
for at least 48 hours to remove residual
oxygen. pH probes were not sterilized after
acid cleaning but were calibrated with pH 7.0
buffer(3-[N-Morpholino]propanesulfonicacid)
and then taken inside the glove box the day
of use.
Barokams were filled while inside the glove
box with brine and nutrients. Filling was
performed in a manner to avoid introducing or
leaving free gas trapped in the barokams.
Once filled, the barokams were removed from
the glove box and fittings tightened with a
wrench before being placed in the oven and
3
connecting to the barokam apparatus. Barokams
were then brought up to 45 C before
pressurizing to the test pressure. In the
process of pressurizing, a small amount of
the barokam contents was pumped through the
outlet to remove any remaining free gas in
the sampling lines, as well as to set the
metering valve at the desired resistance.
Headspace experiments: Glass serum bottles
(120 em3, Bellco) were acid cleaned and
rinsed with deionized water. Bottles and
butyl rubber stoppers were then sterilized by
autoclaving for 15 minutes at 112 C. After
cooling, the bottles were placed in an
anaerobic glove box and preincubated for 48
hours before using. Bottles were filled with
100 cm3 of unfiltered brine and nutrients
leaving approximately 20 cm3 of headspace.
Bottles were stoppered and crimp sealed with
aluminum caps and then placed in a 45 C
incubator housed inside the glove box.
pH measurement: Hydrogen ion concentration
(pH) was performed using pressure tolerant pH
probes (TBI-Bailey Controls, Carson City,
NV). Each barokam pH probe was connected to
a pH Vision Datalogger (Model 6091, Jenco or
Markson Electronics) which recorded pH every
one to three hours to a thermal printer.
Anaerobic glove box: Two types of gloves
boxes were used in these experiments: either
a Coy chamber (Ann Arbor, MI) or a Forma
Scientific Anaerobic System, Model 1024.
Anaerobic gas used in both chambers was
comprised of 10% hydrogen, 5% carbon dioxide,
and the balance nitrogen.
Nutrients and inoculum: Nutrients used in
microbial growth experiments consisted of a
phosphorus source in the form of a phosphate
compound and a carbon source in the form of
a starch hydrolysate.
The microbial inoculum (i.e . indigenous
microorganisms) c o n s i s t e d o f u n f i l t e r e d T r a c t
5 brine. Brine was collected as described
above. In all cases the inoculum was used
within two days of collection. While not in
use the inoculum was stored inside an
anaerobic glove box at room temperature.
Carbohydrate analyses : The analysis of the
starch hydrolysate was performed by adding
 THE EFFECT OF IN SITU PORE PRESSURE ON MEOR PROCESSES
0.1 cm3 or less of sample containing Tract 5
brine to 0.1 cm3 of acetate buffer (0.1 M
( .001 kmol/m3) acetate, pH 4.8 - 5 .O) and 0.1
cm3 of 0.2 M (.002 kmol/m3) KC1. This was
brought up to 0.5 cm3 with deionized water
and .005 cm3 of amyloglucosidase (10 g/L,
Boehringer Mannheim). This solution was
incubated at 45 C for at least 3 hours. At
the end of this incubation the hydrolysate
was assayed for glucose using a glucose
oxidase kit(Sigma Chemical Co, St. Louis, MO)
and the absorbance at 450 nm read with a
Bausch and Lomb Spectronic 21
spectrophotometer. Quantitation was
determined from standard curve analysis
generated with a standard solution of the
starch hydrolysate. Precision and accuracy of
this method was typically between 5 and 10
percent.
Fatty acid analysis: Fatty acid analysis for
acetate, formate, propionate, butyrate, etc.
was performed using quantitative ion
chromatography. Samples were pre-filtered
with OnGuard-Ag filters (Dionex) to remove
chloride. Chromatography of acids was
performed using a Dionex Ion Chromatograph
(Model 4500i) equipped with a Dionex Ion-Pac
ICE-AS1 ion exclusion column and a
conductivity detector. Identification of
acids was performed by comparison to
retention times for authentic standards.
Detection limit was 0.5 ppm.
Fatty alcohol analysis: Fatty alcohols
analysis for methanol, ethanol, acetone,
isopropanol, n-propanol, n-butanol,
isobutanol, sec-butanolandtert-butanolwere
analysed by gas chromatography (Hewlett
Packard GC, Model HP5880) and detected using
a flame ionization detector. Separation of
alcohols was performed on a Carbowax 1500
packed column. Limit of detection was
approximately .001 g/L with a relative
percent standard deviation of 5.
Identification of alcohols was by comparison
of retention times to authentic standards.
Description of coreflood apparatus:The core-
flood apparatus used in all experiments is
depicted in Figure 3. A Ruska positive
displacement pump was used to inject fluids
to drive pistons enclosed in stainless steel
accumulators containing either nutrients,
unfiltered brine, filtered brine, or crude
SPE 24203
oil on the opposite side of the pistons. The
pistons would then drive the solutions
through 1/16" (15.9 mm) ID SS tubing then
through a manifold and into an epoxied core
housed within a SS core holder (annulus) . The
pump oil used to drive the pistons was
degassed with nitrogen that was passed
through a heated copper oven in order to
remove trace amounts of oxygen. It had been
previously established that oxygen in the
pump oil could diffuse from the inside walls
of the accumulator and into the nutrients or
brine. Oxygen would case precipitation of
iron in the brine and is toxic to certain
anaerobic bacteria contained in the brine.
The annulus of the core holder was filled
with water and pressurized above the
injection pressure by means of a hand
operated hydraulic pump. The core holder was
mounted in a forced draft oven held at 45 C.
Validyne differential pressure transducers
(P1 - P4) were mounted along the length of
the core to monitor changes in resistance
factor. These pressures were digitized and
logged by a computerized data acquisition
system. At the outlet end of the core the
stream passed through a pressure tolerant pH
probe (TBI-Bailey Controls, Carson City, NV)
and then through a backpressure regulator set
for 500 psig (3.45 MPa). From here the
effluent would pass through another on-line
pH probe at atmospheric pressure and then
through a gas trap which consisted of a test
tube filled with acidified, oxygen-free water
to reduce carbon dioxide solubility. The
effluent stream was then collected by a
fraction collector or batched into a flask.
Cores:The core used in this study was cut
form preserved Burbank cores. The core was
cut transversely across a larger core (3.5
inch (88.9 mm) diameter) and then cleaned by
solvent extraction in a Soxhlet extractor
using toluene and methanol. The core was
fitted with Ryton endplates and then coated
with epoxy (Dexter Epoxy-Patch). Holes at
approximately 0.5, 1.5, and 2.5 inches (12.7,
38.1, and 63.5 mm) from the inlet face of the
core were drilled into the epoxied rock with
an 1/8 inch (3.2 mm) masonry bit to a depth
of 1/4 inch (6.4 mm). Stainless steel unions
(1/16 inch (1.6 mm), Swagelok) were then
epoxied into these holes and fitted with
stainless steel tubing (1/16 inch (1.6 mm) ,
ID). These taps, designated RF1 through RF4,
SPE 24203 G. E. JENNEMAN AND J. B. CLARK
were used to record differential pressures
across their respective sections of the core
(i.e. RF1, inlet face to 0.5 inches (12.7
mm) ; RF2 0.5 inches (12.7 mm) to 1.5 inches
(38.1 mm) , etc. ) . The core was placed in a
water filled accumulator (annulus) equipped
with a pass through fitting that would allow
the passage of 1/16 inch (1.6 mm) tubing. The
annulus was pressurized such that it' s
pressure was greater than the injection
pressure (approx. 500 psig (3.45 MPa)) being
used. After checking for leaks the core was
vacuum saturated with filtered Tract 5 brine
and the effective permeability to brine
determined. The core was then injected with
clean NBU crude oil and flooded to immobile
water saturation. Afterwards the core was
flooded with brine to residual oil saturation
and the permeability determined.
The resistance factor (RF) is defined as the
permeability to brine after waterflooding to
residual oil saturation divided by the
pernieability to brine after nutrient
injection.
RESULTS
Results of headspace samples are presented as
the mean and standard error of the mean for
four replicates sampled on the same day from
a single source (Figure 4). At 0 psig, 500
psig, and 1000 psig (0, 3.45, and 6.89 MPa)
the mean and standard errors are for sample
sizes of 8,4, and 4 samples, respectively.
These samples were taken on at least two
sample times but from the same sample source.
Additional samples not shown include results
from barokams at 50, 150, 700, 750, 900,
1100, and 1300 psig (.34, 1.03, 4.83, 6.21,
7.58, and 8.96). The results of pH kinetics
for these barokams were similar to those
displayed in Figure 4 for barokams at 0 and
1000 psig (6.80 MPa). In all cases the same
concentrations of starch hydrolysate and
phosphate were used. The results indicate
that the lag time until pH decline is shorter
in those cases where headspace is minimized
and that the rate and magnitude of the pH
drop is greater with increasing pressure.
Figure 5 compares the pH results of two
barokams, one at no hydrostatic pressure and
5
one at 750 psig (5.17 MPa) , in terms of the
difference between pH measured at ambient
conditions (i.e. atmospheric pressure) and
that measured at the test pressure. This
difference should reflect the importance of
increased solubility of biogenic C02 on the
observed lowering of pH since this pH
difference should only reflect the re-
equilibration of the gas (i.e. C02) in
solution with the atmosphere. A value greater
than 1.0 should indicate that there is an
effect of C02 on pH and the greater this
value the greater the effect. A positive
effect is noticed throughout the time period
measured. However the magnitude of the effect
varies with incubation time and the effect is
greater under pressure than at headspace
limiting conditions (i.e. 0 MPa).
Carbohydrate Utilization Rates
In Figure 6 microbial utilization rates for
2.0 g/L of the starch hydrolysate are
displayed as a ratio of the effective
concentration divided by the initial
concentration vs headspace (HS) and
increasing hydrostatic pressure. The solid
bars represent the means and standard errors
of these instantaneous rates calculated at 8
or 9 days from at least four replicates. The
samples are representative of at least two
sampling times from the same sample source.
The exceptions to this are the values at 1100
psig (7.58 MPa) and 1300 psig (8.96 MPa)
which are single sample points taken during
the same sampling time. The asterisk above
the results at 1000 psig (8.96 MPa) indicate
that this mean value was found to be
significantly different from the headspace
sample at the .05 level. Tests of
significance between the mean of the
headspace sample and other means were not
found to be significant. Statistical
significance was determined using the
independent t-test for unequal sample sizes
as performed by Sigmaplot V 4.0 (Scientific
Graph System, Jandel Scientific, Corte
Madera, CA).
Figure 7 compares the utilization rates at
1.0 g/L of the starch hydrolysate for samples
run with headspace and those at 500 psig
(3.45 MPa) of static pressure. These values
represent the means and standard errors of
instantaneous rates measured after 10 or 11
6 THE EFFECT OF IN SITU PORE PRESSURE ON MEOR PROCESSES
days for at least 5 replicates. At least two
sampling times from a single sample source
were used. The mean value at 500 psig (3.45
MPa) was found to be significantly different
(P<.05) from rates determined for headspace
samples. The same statistical methods were
used as for the samples containing 2.0 g/L of
carbohydrate.
Low molecular weight alcohols and fatty acids
in additions to methane, hydrogen and carbon
dioxide comprise the major fraction of
extracellular metabolic end-products formed
through anaerobic fermentation of
carbohydrate feedstocks. Table 2 lists the
effect of headspace and hydrostatic pressure
on the production of the non-gaseous end-
products. The values represent means (S.E.)
of six replicates at headspace conditions,
six replicates under no pressure (i.e. 0
MPa) , and six replicates between the range of
750 - 1000 psig (5.17 - 6.89 MPa) . All ratios
reflect instantaneous measuremenrs and were
normalized for the amount of carbohydrate
utilized at this time (i.e. 14-15 days). No
significant differences were observed for
ratios of ethanol, formate, or acetate under
the various conditions. However, there was a
20 to 35 fold increase in the amount of
methanol in headspace limited samples and in
samples at 750 - 1000 psig (5.17 - 6.89 #Pa)
over the amount found in headspace containing
samples.
In a barokarn experiment conducted for a
longer time period (i,e. 21 days) it was
found that increasing hydrostatic pressure
favored increasing amounts of ethanol per
carbohydrate utilized while acetate per
carbohydrate utilized decreased (Figure 8).
The greater than sign (">") above the bar
indicates that the actual value is greater
than this but due to a limitation in
sensitivity of the carbohydrate assay the
actual value could not be determined. The
less than sign ("<") conversely indicates
that the actual values are less than
indicated.
Core Flood Experiment
A 2.5 X 7.4 cm core plug was drilled,
cleaned, and epoxied as instructed in the
SPE 24203
Materials and Methods section. This core plug
was then placed in the coreflood apparatus
depicted in Figure 3. This core was drilled
from a larger core taken from injection Well
41828 at the North Burbank Unit, operated by
Phillips Petroleum Company. The absolute
permeability of the core to nitrogen gas is
435mD and the effective permeability to brine
is 396mD. The core has an effective porosity
of 26% and a pore volume of 9.8 cc. At
residual oil saturation the effective
permeability to brine is 236mD.
The core was started out with no back
pressure (i.e. zero pore pressure) and at a
temperature of 45 C. The core was inoculated
with approximately 50 pore volumes of
unfilteredTract 5 injectionbrine containing
about 100,000 cells/cm3. After inoculation
the core was injected with growth supporting
nutrients (i.e 2.0 g/L of the starch
hydrolysate and 4.68 mM (4.68 X 10 E-05
kmol/m3) phosphorus (as inorganic phosphate)
at a flow rate of 2.57cm3/h (2.57 X 10 E-05
m3/h). Resistance Factors (RF) were recorded
at specified distances (RFl,RF2,RF3,andRF4)
across the length of the core and
instantaneous readings plotted every four
hours as shown in Figure 9. After
approximately three days (PV = 130) of
nutrient injection the RFs across the entire
-
core began to fluctuate slightly. By the
tenth day (PV 150) the RFs were oscillating
wildly across the core (Figure 9). Pressure
fluctuations across the front end of the core
(i.e. RF1) were not that variable either due
to the increased pressure at the face of the
core and the absence of a free gas phase.
This general behavior continued for the next
nine to ten days at which time the pore
pressure of the core was raised to 500 psig
(3.45 MPa) . Very quickly the pressures
stopped oscillating and became steady. It
took approximately 5 cm3 of brine to compress
the gas in the core to bring it to a pore
pressure of 500 psig (3.45 MPa). Therefore as
much as 50% of the pore volume of the core
could be accounted for as free gas.
DISCUSSION
Effect on vlj
It is a well established fact that the
hydrogen ion concentration has a significant
SPE 24203 G. E. JENNEMAN AND J. B. CLARK
effect on microbial growth and metabolism.
The majority of known microorganisms prefer
a neutral or slightly alkaline environment
but growth and survival at extremes of pH
(i.e. < 1.0 or > 10.0) are known (Langworthy
(1978)). The pH of a medium not only effects
the rate and extent of microbial growth but
effects the types of metabolites formed. A
good example of this is the effect of pH on
the production of acids and solvents (e.g.
ethanol, acetone) by Clostridium
acetylbutylicum where a drop in pH shifts the
production of end-products from acids to
solvents (Gottwald and Gottschalk (1985)).
Therefore pH buffers are often used in
biological media to control pH. The most
natural buffers known are carbonates which
are in equilibrium with bicarbonate, carbonic
acid, carbon dioxide and water. Carbonates
are present in many oil reservoirs as either
rock matrix or as cementing material (i.e.
calcite) which act to hold grains of sand
together. The effectiveness of carbonates as
buffers relies on the formation of the weak
acid H2C03 which in the presence of acid
decomposes to carbon dioxide and water. The
CO, escapes to the atmosphere preventing
accumulation of hydrogen ions. However, the
increased solubility of CO, due to increased
hydrostatic pressure or elimination of
headspace acts to increase the acidity of the
brine by preventing C02 volatilization and
therefore stabilizes the formation of
carbonic acid. In Figure 4 the measured
effect of pressure and headspace on pH
suggests that hydrogen ion concentrations
measured in the presence of a headspace could
underestimate the actual hydrogen ion
concentration by as much as one pH unit or a
factor of 10. Furthermore, in the absence of
a headspace and pressure the pH more closely
follows pH measured at 500 psig (3.45 MPa) or
1000 psig (6.89 MPa) ; however, the results
are highly variable possibly due to an
inability to completely eliminate the
headspace in all the barokams.
Carbon dioxide as well as acidic end-products
(e.g. acetic acid, propionic acid, etc.) of
bacterial fermentation can both affect pH;
however, the importance of these two
effectors could vary throughout the time
course of the fermentation. Figure 5 suggests
that at time zero much of the pH difference
measured at 0 gauge pressure versus 750 psig
7
(6.21 MPa) is due to CO, production or
solubility . This is understandable since the
barokams were initially equilibrated with the
atmosphere inside an anaerobic glove box
which contains 5.0% GO,. Within a couple of
days this CO, effect is lessened possibly due
to microbial fixation of this gas. However,
as the pH as measured under pressure begins
to drop (Days 3 to 5), the pH difference
begins to increase again which can be
explained as oxidative decarboxylation of the
carbohydrate by the fermentative bacteria.
The magnitude of this effect is greater under
pressure presumably due to increased
solubility of CO,. By day 7 or 8 the pH
effect due to C02 begins to lessen but the
measured pH does not increase; therefore the
ability of the medium to maintain the low pH
must be due to production of less volatile
products such as acetic acid. Both acetic
acid and formic acid were detected as end-
products by this stage of the fermentation
under all conditions tested. The dependance
of the pH difference on these fatty acids in
the latter stages of the fermentation could
explain why the pH curves in Figure 4 begin
to converge after 7 or 8 days of growth.
Effect on carbohvdrate utilization
Rates of carbohydrate utilization at
concentrations of 2.0 g/L, measured under
headspace conditions, were significantly
lower than rates that were measured at 1000
psig (6.89 MPa) and greater (Figure 6). At
lower carbohydrate concentrations 1.0 g/L
significant increases in rates were realized
at pressures as low as 500 psig (3.45 MPa,
Figure 7). In some cases, especially in the
presence of headspace, as much as 10 days
elapsed before any significant depletion of
carbohydrate was realized whereas in other
cases utilization began within a couple of
days and proceeded slowly. However under
pressures of 500 psig (3.45 MPa) or greater
utilization began within 5 days and proceeded
at a more rapid rate almost without
exception. The reasons for this increased
rate of utilization are not known. Possible
explanations for these observations are that
pressure selects for a group of
microorganisms with a greater ability to
utilize the carbohydrate or that pressure
enhances the enzymatic activity of the same
population of microorganisms. This effect
8 THE EFFECT OF IN SITU PORE PRESSURE ON MEOR PROCESSES
could take place at the level of the active
site of the enzyme or at genetic loci
controlling the expression of these enzymes.
Pressures in excess of 1000 psig are known to
increase biological reaction rates (Cowen
(1989), Bernhardt et al. (1988), Yayanos
(1975)); however, effects of this type have
not been reported for the relatively low
pressures reported in this study. In fact
very few reports have even examined
biological effects at low pressures. Another
possibility concerns the favorable effect of
pressure on increasing gas solubility which
may select for a microbial consortia with an
increased ability to utilize the
carbohydrate. Samuelov et al. recently found
that increased C02 concentrations increased
the growth rate and decreased the growth lag
phase of the glucose fermenting anaerobe
Anaerobiospirillum succiniciproducens. Also
increased hydrogen concentrations are known
to favor hydrogen consuming (1.e.
hydrogenotrophs) bacteria such as methanogens
and mixotrophs. Hydrogenotrophs can act
synergistically with hydrogen producing
bacteria (i.e. proton-reducing bacteria) in
order to allow a more thermodynamically
favorable fermentation of carbohydrates to
occur (Schink (1988)).
End-products are important in the development
of MEOR processes since it is the production
of these acids and solvents that may be
partially responsible for oil displacement.
Therefore conditions controlling their
production must be evaluated and understood.
The major end-products formed from the
fermentation of the starch hydrolysate in
these experiments were acetate and ethanol.
No significant differences were observed in
the production of these compounds after 14 to
15 days when normalized for the amount of
carbohydrate consumed (Table 2). However for
longer fefmentation periods (i.e. 21 days)
increasing pressure was observed to favor
ethanol production vs acetate production
(Figure 8). This might be expected since
increasing pressure will favor increased
hydrogen solubility which inhibits the
ability of the proton-reducing acetogenic
bacteria to oxidize low molecular weight
fatty acids causing a diversion of electrons
to flow to the more thermodynamically
SPE 24203
favorable reduction of acetate to ethanol.
This suggests that the hydrogenotrophic
methanogens responsible for reducing these
hydrogen levels are rate limiting at this
stage of the fermentation. No attempt was
made to isolate these methanogens and no
attempt was made to measure hydrogen
concentrations in these barokam experiments.
No reports to our knowledge have been made as
to the isolation of hydrogenotrophic
methanogens under the NaCl concentrations
found in the brine under study.
Another observation was the appearance of
significant methanol concentrations produced
when headspace was limited and pressure
increased. Methanol is not considered to be
a major end-product of . anaerobic
fermentations although it is considered a
good substrate for methanogenic bacteria in
hypersaline environments. However, Schink and
Zeikus (1980) reported the production of
methanol formed from the methoxyl groups of
pectin under fermentative conditions. It is
possible that the starch hydrolysate used in
this study contains some methoxylated uronic
acids as impurities. Perhaps an increase in
pressure favors the expression or activity of
the methylesterase responsible for methanol
production, however, no attempts were made to
measure this activity.
Effect of Dressure on ~ermeabilitv
One mechanism that has been proposed for MEOR
is permeability modification of highly
stratified reservoirs which relies on the
selective reduction of permeability in the
high permeability strata. This reduction is
achieved through the production of biomass,
including polymer and cells, within the pores
of the matrix. Another product of microbial
metabolism, gas, can also have an effect on
permeability. The effective permeability of
the rock to brine decreases as the percentage
pore space occupied by gas increases.
Therefore in the absence of pore pressure
biogenically produced gas will occupy more
and more pore space as the gas saturation of
the brine is exceeded. Also the lowering of
pH due to biologically praduced acids will
release more gas as COz. This gas unless
compressed by the in situ pore pressure, will
generate resistances to flow that will
overestimate the actual resistance in situ.
SPE 24203 G. E. JENNEMAN AND J. B. CLARK
The results in Figure 9 demonstrate how a
biogenically produced gas phase effects
resistance factors. As more and more gas is
produced inside the core the differential
pressures fluctuate wildly due to periodic
moments of gas compression and decompression.
The effects are not as noticeable at the
front end of the core where the pressure is
higher and consequently more of the gas is in
solution. When a pore pressure of 500 psig
(3.45 MPa) was applied the differential
pressure and thus resistance factors
stabilized across the core. In the case of
this one coreflood as much as 50% of the
plugging could be explained due to free gas.
Therefore accurate estimates of permeability
reductions due to biomass must be measured at
the in situ pore pressures of the reservoir
under study.
CONCLUSIONS
MEOR experiments should be evaluated
under actual reservoir conditions,
including pore pressure.
Hydrostatic pressures of less than 9.0
MPa can effect pH, substrate utilization,
end-product formation and biogenic
permeability reduction. The presence or
absence of gas headspace can also have an
effect.
Pressure effects are likely a result of
selection of specific microbes and
metabolic pathways that are favored by
the increased solubility of gases.
Additional experiments need to be
performed to determine actual
meehanism(s).
Biogenic gas can have significant effects
on resistance factors measured on cores
run at atmospheric pressures.
ACKNOWLEDGEMENTS
The authors would like to acknowledge the
assistance of Jim Stevens, Deborah Langley
and Everett Johnston in setup and design of
equipment. Also a special thanks to Jim
Stevens, Sheryl Baughman, and Debbie Walker
for help in preparing this manuscript. We
9
also appreciate the opportunity and support
provided by Phillips Petroleum Co.
REFERENCES
Bernhardt, G., R. Jaenicke, H. D.
Ludemann, H. Konig, a n d K . 0. Stetter,
"High pressure enhances the growth rate
of the thermophilic archaebacteriurn
Methanococcus thermolithotrophicus
without extending its temperature
range", Appl. Environ. Microbiol., Vol.
54, pp. 1258-1261 (1988).
Cowen, J. P., "Positive pressure effect
on manganese binding by bacteria in
deep-sea hydrothermal plumes", Appl.
Environ. Microbiol . , Vol . 55, pp. 764-
766 (1989).
Gottwald, M. and G. Gottschalk, "The
internal pH of Clostridium
acetobutylicum and its effect on the
shift from acid to solvent formation",
Arch. Microbiol. Vol. 143, pp. 42-46.
Heinritz, B. , M. Gehrhardt, F. Baumann,
G. Rogge, R. Hedlich and M. Ringfeil,"
Biomass production by thermophilic
microorganisms simultaneously using
reaction heat", J. Chem. Tech.
Biotechnol. Vol. 49, pp. 285-295 (1990).
Jenneman, G. E. "The potential for in-
situ microbial applications", in E. C.
Donaldson, G. V. Chilingarian, and T. F.
Yen (eds.), 'Microbial Enhanced Oil.
Recovery, Elsevier Science Publ. ,
Amsterdam, pp. 37-74 (1989).
Johnson, F. H. and I. Lewin, "The
influence of pressure, temperature, and
quinine on the rates of growth and
disinfection of E. coli in the
logarithmic growth phase, Journal of
Cellular and Comparative Physiology,
Vol. 28, pp. 77-97 (1946).
Langworthy , T . A. "Microbial life in
extreme pH values", in D. J. Kushner
(ed) w
icr bia tr me
Environments, Academic Press, Inc.,
London, pp. 279-315 (1978).
 THE EFFECT OF IN SITU PORE PRESSURE ON MEOR PROCESSES SPE 24203

Marquis, R. E., "High-pressuremicrobial

physiology", Advances in Microbial
Physiology, Vol. 14,pp. 159-241 (1976).

Marquis, R. E. and P. Matsumura,

"Microbial life under pressure", in D.
J. Kushner (ed.) , Microbial Life in
Extreme Environments, Academic Press,
Inc. London, pp. 105-158 (1978).

Samuelov, N. S., R. Lamed, S. Lowe, and

J. G. Zeikus, "Influence of C02-HC0,-
levels and pH on growth, succinate
production, and enzyme activities of
Anaerobiospirillum succiniciproducens",
Appl. Environ. Microbiol., Vol. 57, pp.
3013-3019 (1991).

Schink, B. "Principles and limits of

anaerobic degradation: Environmental and
technological aspects", in A. J. B.
Zehnder (ed.), Biologv of Anaerobic
Microorganisms,
- John Wiley and Sons,
Inc., New York, pp. 771-846 (1988).

Shink, B. and J. G. Zeikus, "Microbial

methanol formation: a major end product
of pectin metabolism. Curr. Microbiol.,
Vol. 4, pp. 387-390 (1980).

Yayanos , A. A. "Stimulatory effect of

hydrostatic pressure on cell division in
cultures of Escherichia coli",
Biochimica et Biophysica Acta, Vol. 392,
pp. 271-275 (1975).

ZoBell, C. E., and L. L. Hittle, "Deep-

sea pressure effects on starch
hydrolysis by marine bacteria", Journal
of the Oceanographical Society of Japan,
Vol. 25, No. 1, pp. 36-47 (1969).
 TABLE 1 TABLE 2

Chemical analysis of injection brine from the Effect of pressure on production of end-products
Tract 5 tank battery at the North Burbank Unit -
after 14 15 days incubation at 45 C.

Concentration
(a/L)
Pressure (g)
ammonium MPa Ethanol Methanol Acetate
nitrate
nitrite Headspace .22(.06) .002(.0006) .13(.02)
sulfate
phosphate
total organic carbon
calcium
barium
magnesium
sodium Values represent averages and standard errors (S.E.)
chloride of the ratios of the concentration of end-products pro-
iron duced divided by the concentration of the carbo-
total dissolved solids hydrate utilized.

To pH Meter
r'
I H
Probe
TO
Sampl i n g
<
\MJ Valve
MGtering
Valve

[~ccumulator

From Reservoir
Fig. l-Barokam.
0 0
anaerobic
Glove BOX
I locc Syringe
9 Meter

Oven
Fig. 2-Barokam apparatus.

n m D
u
a-
OlYl
a*,E n r a n

I
IY1
IU Fig. 3-Coreflood apparatus.
 A Headspace
D p(g) = 0 kPa
v p(g) = 3.45 MPt
0 p(g) = 6.89 MP(

5 10 15
Time (d)

5 10
Time (d)
Fig. 5-ElI.st oi p m u n on pH m w u n d undwr anblmt wnditions vs. musund undar p w s u n .
 Pressure (g) MPa
Flg. GEffect of pressure on the uWlMion of 2.0 g L starch hydmlyast. alter 8-9 day. at 45%.

Headspace 3.45
Pressure (g) MPa
Rs. 7--Etlecl of pressure on tha utiliution of 1.0 plL atamh hydmlyute after 10-11 day. at 45%.
 Ethanol
0Acetate

Pressure (g) MPa

Fig. 8-Elleel of pr-ure on amwnt of end-product formed per carbohydrate u U l W afler 21 days at 45%.

10

0.1
150 200
Pore Volumaa Injected

Fig. 9-The efleel of lnr8u pore prawuw on resistance factorsof a North Burbsnk cow afler mlcmblelplugging.