SPE 126104

New Incubated Pseudomonas Aeruginosa Bacteria for Increasing Oil Recovery

under Reservoir Conditions
M. Samir, SPE, Scimitar Production Egypt Limited; M. Abu El Ela, SPE, WorleyParsons Engineers Egypt Limited; and
S. El Marsafy, S. El Tayeb, SPE, A. Abdel Waly, SPE, and M. H. Sayyouh, SPE, Cairo University

Copyright 2010, Society of Petroleum Engineers

This paper was prepared for presentation at the SPE North Africa Technical Conference and Exhibition held in Cairo, Egypt, 14–17 February 2010.

This paper was selected for presentation by an SPE program committee following review of information contained in an abstract submitted by the author(s). Contents of the paper have not been reviewed
by the Society of Petroleum Engineers and are subject to correction by the author(s). The material does not necessarily reflect any position of the Society of Petroleum Engineers, its officers, or members.
Electronic reproduction, distribution, or storage of any part of this paper without the written consent of the Society of Petroleum Engineers is prohibited. Permission to reproduce in print is restricted to an
abstract of not more than 300 words; illustrations may not be copied. The abstract must contain conspicuous acknowledgment of SPE copyright.

Abstract

Introduction
Many species of microorganisms can produce effective products similar to those described for chemical and miscible EOR
processes. These products can assist in the release of oil from the capillary pores, can improve the sweep and displacement
efficiencies, and increase the oil recovery.1 Some microorganisms produce gases such as carbon dioxide, nitrogen, hydrogen and
methane that could improve oil recovery by increasing pressure and by reducing the viscosity of the crude oil. Some other species
produce acids that can improve the permeability of the reservoir rocks and thus improve the oil recovery. Other microorganisms
produce bio-surfactant that could decrease surface and interfacial tension between oil and water, which causes emulsification and
miscibility and thus improve the oil recovery.
Although several attempts have been made to describe the MEOR process, no pilot test, field trial, experimental model has yet
fully incorporated all of the factors that strongly affect the mechanisms of oil displacement, growth and transport of bacteria in
porous media.2-20 MEOR is a successful process; however, it is limited to specific conditions of low temperature and salinity.
In a pervious work21, the isolation of indigenous bacteria, from three different Egyptian oil fields, succeeded. Three different
species were isolated and identified: Pseudomonas Aeruginosa, Pseudomonas Fluerescens Biotype G and Celluiosimicrobium
Celluians. The previous displacement tests showed that the best species, suitable for MEOR, is Pseudomonas Aeruginosa. A
flooding run was performed on a sand pack model, with the cyclic injection of the bacterial solutions. It was found that the
recovery factor is increased by about 20% when Pseudomonas Aeruginosa is used as it produces biosurfactant and gases. 22
Nutrient with molasses base was found to be the best type of nutrients for this type of bacteria but its activity is largely affected by
the increase in temperature and salinity. 23 In another study24, a new technique was proposed for incubation of the above mentioned
bacteria species and succeeded to increase its efficiency and enable it to withstand the harsh reservoir conditions of high
temperature and high salinity.
The main objective of this work is to study the effect of different reservoir conditions such as lithology, temperature and
salinity on the behavior and activitiy of Pseudomonas Aeruginosa during the displacement process.
2 SPE 126104

Species of Bacteria and Media

The new isolated bacteria “Pseudomonas Aeruginosa” were used to perform the different displacement runs. Pseudomonas
Aeruginosa is the bacterial species isolated from crude oil produced from an Egyptian oil field, located in the Gulf of Suez. 21
This bacterium has the following characterization: round, granular, entire, flat, colorless, translucent, anaerobic, gram-
negative, short rods, performing no spore and acids irresistant. 21 In a previous work, it was found that this type of bacteria has the
ability to produce bio-gases, bio-surfactant and acids. 22 It decreases the viscosity of the oil and the surface tension of the aqueous
and oleic solutions. Also, these bacteria succeeded to increase the oil recovery factor from a sand pack model by around 20% when
using molasses base nutrient.
A "modified media"25 was selected to incubate Pseudomonas Aeruginosa and to improve its growth rate. The composition of
this modified type is given in Table 1.
It was found that the best nutrient type for Pseudomonas Aeruginosa includes the following composition:23 Molasses (2 gm/lit),
NaCl (5 gm/lit) and KNO3 (10 gm/lit).Two brine solutions were used (100,000 ppm and 40,000 ppm). Both of them are obtained
from the previously mentioned Egyptian oil field. The compositions of these two brine solutions are, also, presented in Table 1.
The crude oil was obtained from the same Egyptian oil field. It is free from any other indigenous bacteria. The characteristics
of the crude oil are summarized in Table 1.

Displacement Procedure
A sand pack, with suitable dimensions to simulate the reservoir conditions, was designed to perform the displacement tests. The
sand pack was connected to a vacuum pump, manometer, water path and compressor.
The permeability of the sand pack was about 800 md, which simulates the value of the reservoir permeability. Other properties
of porosity, initial water saturation, OOIP were found to be in the average of 38%, 9% and 360 cc, respectively. All properties were
measured prior the displacement tests. The variation in the rock properties was within a very narrow range which gives confidence
for the reproducibility of the displacement tests. The sand pack was surrounded by hot water to simulate the reservoir temperature.
The procedures of the displacement were as follows:
1. The sand pack is flushed with water of salinity 40,000 ppm until no more oil is produced.
2. About 5 PV of water is injected to reach the residual oil saturation conditions.
3. Then, one PV (first cycle) of the bacterial solution is injected; and the sand pack is left for one week for incubation.
4. About 3 PV of water is injected until no more oil is produced.
5. Another one PV of bacterial solution (second cycle) is injected and the sand pack is then left for another one week for
incubation.
6. The sand pack is flooded again with water.
7. The properties of the effluent fluids are then measured.

Analysis of Results

Effect of Temperature. The effect of applying MEOR process on the ultimate recovery can be observed in Figs. 1, 3, 5 and 6. It is
clear from these figures that applying the MEOR process after the waterflooding increased the oil recovery by about 20%.
Figs. 1 and 2 reflect the effect of incubation temperature on the behavior and activity of the used bacteria “ Pseudomonas
Aeruginosa”. As shown in Fig. 1, there is a slight increase in the ultimate recovery factor (about 2%) when the incubation
temperature of the bacteria is increased from 50ºC to 70ºC. This result indicates that the behavior and activity of the used bacteria
are not affected when the incubation temperature increases from 50 to 70ºC. The small increase in the oil recovery at the higher
temperature can be related to the effect of temperature on the oil viscosity. The viscosity of the produced oleic phase was
measured. It was found that there is a slightly decrease in the viscosity with increasing the temperature: (from 28.3 cp at 50°C to
27.5 cp at 70ºC). Reduction of the oil viscosity with temperature improves the oil mobility and the mobility ratio, and increases the
amount of oil recovered by displacement.
The properties of the effluent fluid confirm also that the behavior and activity of the used bacteria are not affected when the
incubation temperature increases from 50 to 70ºC. The surface tension of the aqueous phase for the effluent fluid at the two
temperatures was almost the same (38 dyne/cm). Fig. 2 indicates that the pH value and conductivity of the effluent solutions, at 50
and 70ºC are almost identical. This implies that the ability of the bacteria to produce the bioproducts at 70ºC is the same as its
ability at the lower temperature (50ºC). These results are considered a reasonable proof to the validity of the new incubation
technique that was developed to increase the ability of the used bacteria to withstand the high temperature conditions of the
reservoir.24
SPE 126104 3

Effect of Formation Water Salinity. The activity and the behavior of the used bacteria at formation water salinity of 100,000 ppm
and 150,000 ppm and temperature of 70ºC were studied. It is clear from Fig. 3 that there is a slight decrease in the recovery factor
by about 4% due to the increase of the salinity from 100,000 ppm to 150,000 ppm. This insignificant reduction in the ultimate
recovery factor indicates that the used bacteria still has reasonable activity and behavior and gives relatively noticeable
performance even at harsh reservoir conditions of high salinity and high temperature: An oil recovery factor of 82% is observed at
a reservoir temperature of 70ºC and salinity of 150,000 ppm.
The properties of the effluent fluid confirm also that the behavior and activity of the used bacteria are slightly affected when
the salinity increases from 100,000 ppm to 150,000 ppm at a temperature of 70 ºC. It was found that there is a slight increase in the
surface tension with salinity (from 38 dyne/cm at 100,000 ppm to 39 dyne/cm at 150,000 ppm). However, the viscosity was almost
the same (27.5 cp).
Fig. 4 shows that the pH value and conductivity of the effluent solutions at 100,000 ppm and 150,000 ppm are almost identical.
This indicates that the activity of the bacteria is almost the same at the two considered values of formation water salinity.
On applying waterflooding, without using bacteria, the ultimate oil recovery factor was found to be about 60%, as illustrated in
Fig. 3. However, activation of the bacteria can increase the oil recovery by about 22%, despite the high formation water salinity of
150,000 ppm and high reservoir temperature of 70ºC.

Effect of Incubation of the Bacteria outside the Sand Pack. The effect of changing the location of incubation of the used
bacteria on the oil recovery factor is demonstrated in Fig. 5. It is clear from this figure that the recovery factor in case of
incubation of the bacteria outside the sand pack is higher than the recovery factor in case of incubation of the bacteria
inside the sand pack by about 5%.
The activity and the performance of the bacteria in the two incubation locations were studied. The surface tension of the
aqueous phase and the viscosity of the oleic phase from the effluent fluids recovered from the sand pack at the two incubation
locations were measured. The surface tension decreased from 38 dyne/cm on incubating the bacteria inside the sand pack to 37
dyne/cm on incubating the bacteria outside the sand pack. Moreover, the viscosity of the oleic phase decreased from 28.3 cp on
incubating the bacteria inside the sand pack to 26.5 cp on incubating the bacteria outside the sand pack.

Effect of Different Lithology. Fig. 6 indicates that the presence of Kailinite and Dolomite in the sand pack does not affect the
final oil recovery significantly. The only difference between the two cases, presence or absence of Kailinite and Dolomite, is that
the final value of the recovery factor is reached faster in the first case (after 4.5 PV), while in the second, it is reached after 6 PV.
This can be related to the change of the permeability due to the presence of the Kailinite and Dolomite.

Conclusions
 The value of the ultimate oil recovery factor increased by about 20%, over its initial value, after the water flooding
process due to the use of bacteria Pseudomonas Aeruginosa.
 On increasing the temperature and salinity up to 70ºC and 150,000 ppm, the behavior and activity of the used bacteria are
nearly the same as its behavior and activity at lower values of temperature (50ºC) and salinity (100,000 ppm).
 Higher recovery can be obtained on incubating the bacteria outside the reservoir.

References
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Annual Technical Conference and Exhibition, Houston, Texas, 3-6 October 1999.
4 SPE 126104

9. Wei, C., et al, "Enhance Oil Production in High Waxy Oil Reservoir by Single Well Cyclic Microbial Injection Production,"
SPE Asia Pacific Improved Oil Recovery Conference, Kuala Lumpur, Malaysia, 25-26 October 1999.
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Asia Pacific Improved Oil Recovery Conference, Kuala Lumpur, Malaysia, 25-26 October 1999.
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Conference, Kuala Lumpur, Malaysia, 25-26 October 1999.
14. Karim, M.G., et al, "Microbial Enhanced Oil Recovery (MEOR) Technology in Bokor Field, Sarawak," SPE Asia Pacific
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Carbon Source," SPE Asia Pacific Improved Oil Recovery Conference, Kuala Lumpur, Malaysia, 8-9 October 2001.
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Oil Recovery Symposium, Tulsa, Oklahoma, 13-17 April 2002.
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Approach," SPE International improved oil conference, Asia Pacific, Kuala Lumpur, Malaysia, 20-21 October 2003.
18. M. Abu El Ela, "Role of Indigenous Bacteria in Crude Oil Reservoirs on Relative Permeability Curves," M.Sc. Thesis, Cairo
University, July2001.
19. Abu El Ela, M., El Tayeb, S., Sayyouh, M., Abdel Dayem, M., and Desokey, S., “ Effects of Stimulating Indigenous Bacteria
in Oil Reservoirs on Relative Permeability Curves,“ SPE 75240, SPE/DOE Thirteenth Symposium on Improved Oil
Recovery, Tulsa, Oklahoma, USA., 13-17 April 2002.
20. Abu El Ela, M., El Tayeb, S., Sayyouh, M., Abdel Dayem, M., and Desokey, S., “Effects of Stimulating Indigenous Bacteria
in Crude Oils on the Recovery,” presented at the Eighth International Conference on Mining, Petroleum & Metallurgical
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2004, Cairo, Egypt.
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SPE 126104 5

TABLE 1- FLUID AND ROCK PROPERTIES

Composition of the Modified Media(25)
Proteose peptone, gm/lit 20
Glycerol, gm/lit 10
K2HPO4, gm/lit 10
MgCl2.6H2O, gm/lit 1.4
Composition of the Sea Water
NaCl, ppm 28,500
CaCl2, ppm 1,500
MgCl2, ppm 3,000
CaSO4, ppm 7,000
Total Salinity, ppm 40,000
Composition of the Formation Water
NaCl, ppm 50,000
CaCl2, ppm 42,000
MgCl2, ppm 8,000
CaSO4, ppm 0
Total Salinity, ppm 100,000
Properties of the Crude Oil
Initial Reservoir Pressure, psi 3000
Reservoir Temperature, ºF 180
Bubble Point Pressure, psi 1350
GOR, SCF/STB 260
Oil Gravity, ºAPI 20
Pour point, ºF 27
Density, gm/cc 0.92
Asphalt content, % 12
Properties of Sand pack Porous Media
Porosity, % 38
Permeability, md 834
Initial Water Saturation (Swi), % 9
Original Oil In-Place, cc 360
Effective Oil Permeability at (Swi), md 705

Fig. 1- Effect of temperature on the produced oil and recovery factor on application of waterflooding and incubating the bacteria inside
the sand pack
6 SPE 126104

Fig. 2- Effect of temperature on the conductivity and pH of the produced aqueous phase on incubating the bacteria inside the sand pack

Fig. 3- Effect of formation water salinity on the produced oil and recovery factor on application of waterflooding and incubating the
bacteria inside the sand pack at 70ºC
SPE 126104 7

Fig. 4- Effect of formation water salinity on the conductivity and pH of the produced aqueous phase on incubating the bacteria inside the
sand pack

Fig. 5- Produced oil and recovery factor on application of waterflooding and incubating the bacteria inside/outside the sand pack at 50ºC
8 SPE 126104

Fig. 6- Effect of different lithology on the produced oil and recovery factor on application of waterflooding and incubating of the bacteria
inside the sand pack at 50ºC