Control of Microbiologically Induced Corrosion in

Arab Journal of Nuclear Sciences and Applications, 45(2)460-478(2012)
Control of Microbiologically Induced Corrosion in Petroleum Industry

Using Various Preventive Strategies
Abeer E. Zakaria*, H. M. Gebreil* * and Naglaa M. Abdelaal*
* Microbiology Department, National Center for Radiation Research and Technology, Egypt.
** Microbiology Department, Ain Shams University, Egypt.
ABSTRACT
Various inhibitive strategies were used to control Sulfate reducing bacteria (SRB).
The traditional treatment of SRB by biocides was compared with other treatments such
as exposure to microwaves, ultraviolet, gamma radiation separately and addition of
either nitrate or nitrite. Six commercial biocides were tested for SRB control.
Champion- A was found to be the most efficient biocide. Addition of nitrate to SRB
growth medium did not inhibit the growth at 10 mM/l whereas addition of 6 mM/L
nitrite completely inhibited the growth. On the other hand, physical treatments achieved
satisfactory results. the lethal doses required for complete inhibition of the growth on
using microwave, ultraviolet and gamma radiations were found to be at (50 second, 4
hours and 0.9 KGy) respectively. On studying the effect of the lethal and sublethal doses
of different treatment on the growth and sulfide production rates of SRB, it was found
that the lethal doses of all studied treatments except nitrate treatment achieved complete
inhibition. Also the corrosion aspects and elemental analysis of metal coupons surfaces
at such doses showed a clear variation in distribution and composition of the corrosion
products adhered to their surfaces.
Key Words: Microbiologically Induced corrosion (MIC)/ Sulfate Reducing Bacteria
(SRB)/ Corrosion Control.
INTRODUCTION
In aqueous environments, iron materials are corroded not only by purely chemical or
electrochemical reactions but also by microorganisms or the products of their metabolic activities
including enzymes, exopolymers, organic and inorganic acids as well as volatile compounds such as
hydrogen sulfide in a process termed Microbiologically Induced Corrosion (MIC) (Miranda et al.,
2006). Booth (1964), in the UK, suggested that 50% of corrosion failures in pipelines involved MIC
and the replacement costs for biocorroded gas mains were recently reported to be 250 million per
annum (Beech and Gaylarde, 1999). Bacterial activity and mainly Sulfate Reducing Bacteria (SRB)
activity responsible for > 75% of the corrosion in productive oil wells and > 50% of the failures of
buried pipelines and cables. It has been reported to be responsible for extensive corrosion of drilling
and pumping machinery and storage tanks (Javaherdashti, 2008). The presence of SRB in oil
environments was rapidly recognized as responsible for the production of hydrogen sulfide, which is a
toxic and corrosive gas responsible for a variety of environmental and economic problems including
reservoir souring (increase sulfur content) (Hubert et al.,2003). It is resulting in decrease the quality
and value of oil, contamination of natural gas and oil, corrosion of metal surfaces, and the plugging of
reservoirs due to the precipitation of metal sulfides in the fluid flow paths and the consequent
reduction in oil recovery (Magot et al., 2000;Nemati et al.,2001; Davidova et al.,2001 and Hubert
460
et al., 2005). So due to the detrimental effects of SRB in the oil industry, they have been the most
commonly studied group (Miranda et al., 2006).
Considerable efforts have been directed toward controlling SRB growth and inhibition
of corrosion induced by its activity. Corrosion inhibition is the slowdown of the corrosion
reaction usually performed by substances which when added in small amounts, they
decrease the rate of attack by these bacteria on a metal (Ash and Ash, 2001). A number of
methods for controlling SRB sulfide production in different oil production facilities
including the use of biocides (Gardner and Stewart, 2002). Although biocide treatments
are widely used to decrease biofouling and MIC in steel pipes and in closed systems, the
results are far from satisfactory. This is because biocides are much less effective against
sessile microorganisms with biofilms compared to their effectiveness against planktonic
populations. In addition, biocide resistance may be developed and biocidal action reduced
by dilution (Gardenr and Stewart, 2002). An alternative approach for the control of SRB
sulfide production in water- oil systems is the use of repeated injection of nitrate
(Thorstenson et al., 2002 and Kjellerup et al., 2005). Also the promise of nitrite addition
as an effective sulfide control strategy was reported by Mohanakrishnan et al. (2008). The
use of nitrate and nitrite was proven to be very effective and was originally attributed to the
inhibition of SRB (Zhang et al., 2008). Also Cirne et al. (2008) reported that addition of
limited amounts of nitrate or nitrite is a simple strategy to control anaerobic sulfide
production by SRB. On the other hand the physical treatment strategy such as ultra violet
(UV), microwave, and radiations provides an alternative method to control SRB as both of
these methods are non intrusive and do not require the injection of any foreign material
(Bjorndalen et al., 2003). The aim of this study is comparing between different strategies
for controlling SRB growth and prevention of corrosion induced by it and selection of the
most efficient strategy among them.
MATERIALS AND METHODS
Sampling and preparation of SRB inoculum
A water sample from Agiba oilfield located in Egypt was collected in sterile and anaerobic
polypropylene bottles, then stored in ice coolers upon collection and analyzed within 24 hours. Ten ml
of water sample was inoculated into a tightly closed anaerobic 500 ml bottle containing a liquid
Starkey growth medium (Starkey, 1948). The inoculum of SRB culture used in all experiments was
prepared by taking 1 ml of stock culture solution and inoculated into a vial containing 9 ml of
anaerobic Starkey liquid medium for 3 days at 33 C.
Enumeration and estimation of sulfide productivity of SRB
The Enumeration of SRB in all experiments was done using the Most Probable Number (MPN)
technique according to ASTM D-4412 (1990).The MPN count of SRB was compared with the
statistical table of Cochran (Cochran, 1950). Differences were considered significant at the 95%
confidence interval. The total dissolved sulfide in the medium was determined spectrophotometrically
according to (Cord- Ruwish, 1985) method.
Evaluation of different biocide efficacy
Six different biocides (Champion A, Champion B, E.C.C. A, E.C.C. B, biocide Z-A and biocide
X-B) obtained from the Egyptian Petroleum Research Institute (EPRI)were evaluated for inhibition of
461
SRB. Each biocide was added to sterile anaerobic vials containing 100 ml of Agiba oilfield water
sample inoculated with 1 ml of SRB culture to give final biocide concentration 100 ppm. Each vial
was inoculated with 1ml of SRB inoculum. The control vial was left untreated to be the control vial.
Each biocide was kept in contact with SRB for 2 hours at room temperature (the contact time). Then
one ml aliquots of each vial were withdrawn by a sterile syringe and inoculated into free biocide
Starkey medium vial and incubated at 33C for 21 days. The MPN values of SRB were recorded. The
biocide which inhibited the SRB growth at 100 ppm will be selected for evaluation of its
efficacy at concentration less than 100 ppm in the next experiment. The selected biocides will
be tested in concentrations (10, 30, 50, 80& 100 ppm). Each biocide was added to sterile
anaerobic vials containing 100 ml of Agiba oilfield water sample inoculated with 1 ml of SRB
culture to give the desired final biocide concentration. One vial was left untreated to be the
control vial. After 2- hours contact time, one ml aliquots of each vial were withdrawn by
syringe and inoculated into free biocide Starkey medium vial and incubated at 33C for 21
days. The MPN values of SRB were recorded.
Evaluation of nitrate and nitrite addition on SRB growth
One ml of SRB enumerated in Starkey medium containing different nitrate (NO3) - and nitrite
(NO2) concentrations (0, 2, 4, 6, 8& 10 mM/l) separately. The inoculated media incubated at 33C for
21 days. The MPN values of SRB and the amount of produced sulfide in each medium were recorded.
Evaluation of different irradiation strategies
One ml of SRB culture was added separately to 9 ml Agiba oilfield water sample in different
sterile anaerobic vials to create the test samples. The test samples were then exposed to microwave
and ultra violet and gamma irradiation separately at doses (20, 30, 40 & 50 second), (1, 2, 3 & 4
hours) and (0.1, 0.3, 0.5, 0.7 & 0.9 KGy) respectively. After irradiation, 1 ml of each sample was
inoculated into Starkey medium. The inoculated medium incubated at 33C for 21 days. The MPN
values of SRB of each sample were recorded.
Preparation of metal coupons for pre and post- test examination
Prior to use, the surfaces of metal coupons were metallographically polished according to
(ASTM G 1-72, 1993). The metal coupons were serially polished with 80, 120, 220 and 500 grit
silicon- carbide papers, degreased in acetone, washed with sterile distilled water. Then coupons were
dried in a current air and sterilized with ethanol before exposure to the experimental media (GonzalezRodriguez et al., 2008). After metal coupons use (at the end of each experiment), the metal coupon
surfaces were cleaned under a stream of tap water and scrubbed vigorously with rubber stopper (Zuo
et al., 2004).
Studying the effect of lethal and sublethal doses of different treatment on both the growth and
activity of SRB and corrosion aspects of metal coupons
One ml of SRB inoculum was inoculated separately into vails containing 9 ml Agiba oilfield
water sample and exposed to the sublethal and lethal doses of champion A biocide, microwave,
ultraviolet and irradiations. An inoculated untreated vial was kept as a positive control. After
treatments directly, one ml aliquots of each treated vial and the control one were withdrawn by syringe
and enumerated in the modified medium at 33C for 21 days to obtain the count at zero time after
treatment. On the other hand, one ml aliquots of each treated vial and the control one was inoculated
into the modified medium vial containing a metal coupon for studying the corrosion aspects of the
metal after exposing SRB to each treatment and incubated at 33C for 30 days. One medium vial
containing a metal coupon was left uninoculated to study the corrosion aspect of the metal coupon in
462
the medium in the absence of SRB (negative control). At regular time intervals upon 30 days, one ml
of the inoculated vials containing a metal coupon was withdrawn and enumerated in another fresh
medium. The MPN values of SRB and the amount of produced sulfide was estimated upon 30
days.For studying the corrosion aspects of the metal coupons surfaces at nitrate and nitrite treatment
strategy, one ml of SRB inoculum was cultured and enumerated separately into two sets of 9 ml
medium vials with a metal coupon, one set containing (4& 6 mM/l) of nitrite and the other set
containing (10 mM/l) of nitrate, then incubated at 33C for 30 days. The MPN values of SRB and the
amount of produced sulfide were estimated upon 30 days at different time intervals. At the end of
experiment (after 30 days), the metal coupons were taken from the vials. Finally the corrosion aspect
and elemental analysis of the metal coupons surfaces were examined by Scanning Electron
Microscope (SEM) coupled with Energy Dispersive X- ray (EDX).
RESULTS AND DISCUSSION
It is concluded from Figure (1) that biocide treatment for two hours contact time sharply decreased
the count of SRB comparing to the control. Biocide X-B exhibited less efficiency than Champion B but
both biocides exhibited reduced count than the control whereas other biocides stopped the growth. So
biocides Champion B and X-B were excluded from the following evaluation studies. Also it was observed
that the all tested biocides of aldhydic (Champion A, E.C.C.A &Z-A) and only one of amine type biocides
(E.C.C.B) exhibited complete inhibition of the growth. Hence, it can be concluded that the aldhydic
biocides were more effective than the amine type biocides for SRB inhibition. These data goes parallel to
what has been achieved by other investigators. Von Reg and Sand (1998) evaluated different biocide
efficacy for SRB treatment. The samples were treated with the biocide formaldehyde,
tetramethylammoniumhydroxide, 1,8 dihydroxyanthraquinone and a commercial biocide named Dilurt at
varying concentrations It was found that formaldyde exhibited the best effect. Only 3% of the original
microbial activity remained and reduction in SRB cell numbers of five orders of magnitude. In contrast,
tetramethylammoniumhydroxide had only slight effect. Microbial activity was reduced only to 20% and the
cell numbers did not decrease at all. The other biocide exhibited intermediate effects. Also Gardner and
Stewart (2002) reported that 50 ppm of glutaraldehyde retarted the SRB growth to 143 hour in Postgate C
medium due to its ability to cross- link proteins.
Fig. (1): Final SRB counts at 100 ppm concentration of different biocides after 21 days of
incubation.
463
From Figure (2), it is clear that Champion A is the most effective biocide among the others. As
at all its concentrations, it exhibited high efficiency for reducing SRB count contrary to others biocides
concentrations. Also it exhibited the lowest minimal inhibitory concentration at (80 ppm) whereas for
others, it was at (100 ppm). So it will be selected as the most efficient biocide among the others. The
modes of action of these biocides may be attributed to their electrochemically active properties as they
react with the nucleophilic components of the microbial cell or their ability to form chelates with metal
cations necessary for cell metabolism (Heitz, 1996). Other mode of action is that the ingredients of
biocide are membrane active which coat the cell wall of the microbes adsorptively. This process
causes changes in the outer membrane. These outer barriers loose their integrity with the results that
the biocide molecules are allowed access to cytoplasmic membrane, so that they can release their
lethal effects, resulted in inhibition of the enzymes localized in the cytoplasmic membrane, escape of
essential components from the cytoplasm (Heitz, 1996).
Fig. (2): The final SRB counts after 21 days of incubation at different concentrations of the four
tested biocides.
The overall effect of nitrate and nitrite addition on the final counts and amounts of produced
sulfide by SRB after 21 days of incubation is illustrated in Figure (3). The data shows that the growth
and sulfide production activity of SRB were completely inhibited at 6 mM/l nitrite whereas at the
same concentration or even at the higher concentration up to (10 mM/l) of nitrate, the count and
sulfide production activity were only reduced without complete inhibition. From this observation, it
can be concluded that nitrite is more effective than nitrate for controlling SRB growth. This conclusion
agreed with some worker's studies. Sturman and Goeres (1999) suggested that the use of nitrite alone
as an alternative to nitrate may promote higher reactivity and more rapid scavenging of sulfide.
Kjellerup et al. (2005) demonstrated that nitrite alone reduced the SRB production of sulfide, while
nitrate alone had no effect. Also Garcia De-lomas et al. (2007) reported that sulfate reduction activity
was not fully inhibited by nitrate addition.
464
On the other hand there were other researches disagreed with this conclusion. Jenneman et al.
(1986) found that the addition of (59 mM/l) nitrate completely inhibited the sulfide production and the
number of SRB decreased with prolonged incubation period. Also Myhr et al. (2002) found that
injection on 0.5 mM nitrate for 2.5- 3.5 months led to complete elimination of H2S.
Nitrite- Count
Nitrite- Sulfide conc.
4.5
4
3.5
3
2.5
0.6
0.5
0.4
0.3
2
1.5
1
0.5
0
0.2
0.1
Sulfide Conc. (mM/L)
Count (Log MPN)
Nitrate- Count
Nitrate- Sulfide conc.
0
0
10
Nitrate/ Nitrite Conc. (mM/l)

Fig. (3): Comparison between the effect of nitrate and nitrite addition on both the final counts
and amounts of produced sulfide by SRB.
Figures (4) represent rates of SRB growth when exposed to different doses of microwave
radiation, the results show that with increasing microwave exposure period, the SRB growth sharply
decreased. It was observed that SRB growth was stimulated by very low exposure doses of microwave
whereas the high doses inhibited it. Also it was cleared up that the lethal dose required for complete
growth inhibition is (50 sec.) of exposure period.
4.5
4
Count (Log MPN)
3.5
3
2.5
2
1.5
1
0.5
0
0
20
30
40
50
Exposure time (second)
Fig. (4): Effect of different microwave exposure doses on the MPN values of SRB after 21 days
of incubation.
465
These results are supported by various examinations of other microorganisms exposed to

microwave irradiation and the inhibitory action of microwaves on SRB growth was reported by
(Fujikawa and Ohta, 1994; Sato et al., 1996 and Hamid et al., 2001). It has also been previously
reported that the destructive effect of microwave irradiation is not only due to temperature change but
also the irradiation effect itself (Sato et al., 1996). Also Bjorndalen et al. (2003) studied the effect of
microwave irradiation on SRB and concluded that with increasing microwave time, the growth of the
bacteria is inhibited. Also he deduced that with slight microwave irradiation, SRB is activated. This
may be attributed to slight molecular agitation of certain molecules such as water produced by the
action of microwave irradiation.
On studying the effect of U.V exposure on SRB growth, it was found from Figure (5) that on
increasing U.V exposure period, there was a significant decrease in the count of SRB. Also the
presented data pointed out that the 4 hours exposure period was the lethal dose which was sufficient
for complete inhibition of SRB growth. The inhibitory effect of ultraviolet radiation can be attributed
to the production of a limited of specific types of chemical changes. The aromatic rings of the purines
and pyrimidine bases selectively absorb ultraviolet radiation where two adjacent pyrimidine in the
same chain (T-T, C-C or T-C) become covalently bonded together; these unpleasant pyrimidine
dimmers disrupt the local structure of the DNA (Eckardt-Schupp and Klause, 1999).
It has been proved that the U.V. radiation method is effective for SRB elimination from ground
water. It was found that doses below 40 mJ/cm2 were ineffective in SRB elimination. In doses ranging
from 40 to 75 mJ/cm2 the effectiveness of SRB removal varied from 55 to 86%. The highest removal
effectiveness (about 100%) was observed when UV doses above 77 mJ/cm2 were applied (Wargin et
al., 2007). Although the efficiency of U.V in a 99% reduction in viable bacterial numbers, its effect
was efficient only at very long exposure time. This may be due to the poor penetrating power of
ultraviolet light (Mittelman, 1990). Some investigators have addressed U.V application as an
alternative to biocides (Saiz-jimenez, 2001).
4.5
4
Count (Log MPN)
3.5
3
2.5
2
1.5
1
0.5
0
0
Exposure time (hour)
Fig. (5): Effect of different UV exposure doses on the final MPN values of SRB after 21 days of
incubation.
466
The results presented in Figure (6) indicated that on increasing gamma radiation doses, the
growth sharply decreases. The control (non-irradiated) sample exhibited the highest SRB count
(2.4x105). At low dose of gamma radiation (0.1 KGy), the growth less final count (2.1x103) than at the
control sample. At doses (0.3, 0.5& 0.7 KGy), the growth exhibited final counts (2.0x10, 7.8& 2.0)
respectively showing a dramatic decrease in the count. Also the given data pointed out that the lethal
dose of gamma radiation required for complete inhibition of SRB growth is at (0.9 KGy). Among the
methods which can be used for changing metabolic activities of living cells is gamma radiation
(Alabostro et al., 1987). Several previous studies recorded that the low doses of gamma radiation may
stimulate the microbial metabolic activities (El-Batal and Khalaf, 2003). Meanwhile, high doses of
gamma radiation were proved to be inhibitory of microbial activities (Meleigy, 2009).
6
Count (Log MPN)
0
0
0.1
0.3
0.5
0.7
0.9
Exposure dose (KGy)
Fig. (6): Effect of different gamma irradiation doses on the MPN values of SRB after 21 days of
incubation.
The growth rate of SRB upon 30 days was determined in the modified medium containing a
metal coupon after the exposure to the lethal and sublethal doses of champion A biocide, microwave,
ultraviolet, gamma radiation, nitrite and the most efficient nitrate concentration for controlling SRB in
order to study the effect of such treatment doses on the growth rate of SRB. All samples were
inoculated with initial count (1.0x 104) of SRB. Figure (7) revealed that the control sample (untreated)
exhibited the highest growth rate and count comparing to the treated samples. The lethal doses of all
the studied treatments as well as the lethal and sublethal doses of Champion A biocide (50& 80 ppm)
respectively exhibited complete inhibition of SRB growth without growth recovery upon 30 days of
incubation.
On the other hand the nitrate treatment (10 mM/l) exhibited relatively reduction in SRB count.
At the second day of incubation, the SRB growth at the control sample (untreated) increased by one
log cycle comparing to the initial count and continued the increase to reach the highest final count
(1.1x106) at day (12), then entered the stationary phase up to day (20). The decline phase appeared at
day (24) at which the count decreased to be (1.8x105). On the other hand at day (2) of incubation, the
nitrate dose (10 mM/l) and the sublethal doses of nitrite (4 mM/l), microwave (40 sec.), U.V. (3 hours)
and gamma radiation (0.7 KGy) sharply reduced the counts by ratio (79, 93, 98, 97& 99%) of the
initial count respectively, whereas the growth at such treatments started to recover and the counts
467
gradually increased to reach final counts (6.6x103, 1.8x103, 3.6x103, 8.1x102& 1.0x102) at days (12,
16, 12, 12& 8) respectively after that the growth entered the stationary phase. Then the counts started
to decrease at the decline phase to be (3.8x10, x& 2.1x10) at the sublethal doses of microwave,
U.V. and gamma radiation respectively at day (30). Whereas at the nitrate dose (10 mM/l) and the
sublethal dose of nitrite (4 mM/l) the growth remained constant. Hubert et al. (2005) found that the
treatment with 17.5 mM nitrate lowered the planktonic SRB by 3-5 log units, whereas, direct addition
of 20 mM nitrite reduced SRB populations by 3-7 log units.
Control
Nitrate(10mM/l)
Microwave (40 sec.)
U.V. (4 hours)
Campion-A(50 ppm)
U.V.(3hours)
Microwave (50 sec.)
Radiation (0.7KGy)
Campion-A(100 ppm)
Nitrite (6mM/l)
Nitrite (4mM/l)
Radiation (0.9KGy)
Count (Log MPN)
6
5
4
3
2
1
0
0
12
16
20
24
26
30
34
Time (day)
Fig. (7): Effect of the lethal and sublethal doses of different treatments on the growth rates of
SRB.
The sulfide production rate of SRB upon 30 days was determined during its growth in the
modified medium containing a metal coupon after the exposure to the lethal and sublethal doses of
champion A biocide, microwave, ultraviolet, gamma radiation, nitrite and the most efficient nitrate
concentration for controlling SRB (10 mM/l) in order to study the effect of such treatment doses on
the sulfide production rate of SRB. Data given in Figure (7) indicated that the lethal doses of all the
studied treatments as well as the sublethal dose of Champion A biocide (50 ppm) exhibited complete
inhibition of the SRB sulfide productivity as the sulfide was absent in the medium upon 30 days of
incubation.The sulfide was immediately detected in the medium at the second day of incubation for
the control sample, whereas for the treated samples with the sublethal doses of nitrite (4 mM/l),
microwave (40 sec.), U.V. (3 hours) and nitrate dose (4 mM/l), it was detectable at day (5). Also it was
observed that sublethal dose of gamma radiation resulted in suppression of the sulfide production
activity for (8) days. Also the presented data revealed that the control sample (untreated) exhibited the
highest sulfide productivity (0.48 mM/l) comparing to the all treated samples. The sulfide productivity
was dramatically reduced by approximately ratio (56, 66, 73, 75& 81%) at the nitrate dose (10 mM/l)
as well as the sublethal doses of nitrite (4 mM/l), U.V. (3 hours) microwave (40 sec.), and gamma
radiation (0.7 KGy) respectively comparing to the control sample. These obtained data agree with
earlier scientific results. The response to nitrate treatment was a rapid reduction in number and activity
of SRB in the water injection system. The activity of SRB has remained low at < 0.3 and < 0.9 g H2S
cm2/ day at Veslefrikk and Gulfaks oil fields respectively (Bodtker et al., 2008).
468
Control
Nitrate(10mM/l)
Microwave (40 sec.)
U.V. (4 hours)
Campion-A (50 ppm)

U.V.(3hours)
Microwave (50 sec.)
Radiation (0.7KGy)
Campion-A (100 ppm)

Nitrite (6mM/l)
Nitrite (4mM/l)
Radiation (0.9KGy)
0.6
Sulfide Conc.(mM/l)
0.5
0.4
0.3
0.2
0.1
0
0
12
16
20
24
26
30
34
Time (day)
Fig. (7): Effect of lethal and sublethal doses of different treatments on the sulfide production
rates of SRB.
From the EDX analysis and SEM images presented at Table (1) and Figure (8), a clear variation
can be noticed between the positive control sample (a coupon immersed in medium inoculated with
untreated SRB) and the negative control (a coupon immersed in medium without SRB) as well as the
samples treated at lethal doses of nitrite, microwave, ultraviolet and gamma radiation (coupons
immersed in medium inoculated with SRB exposed to the applied treatments separately). The SEM
images showed many corrosion pits on the metal surface of the positive control coupon and non
contentious deposits of bacteria adhered to the surface were clearly observed (Fig. 8-B). Whereas, the
negative control showed a clear surface without pits or any deposits on the surface (Fig. 8-A).
The previous data were confirmed by the EDX elemental analysis of the metal surface and
represented in Table (1) and Fig. (8-A, right column); it shows a clear variation in distribution and
composition of the corrosion products between the positive control and the negative one. There was an
intense accumulation of sulfur based compound (may be iron sulfide) on the surface of metal coupon
of the positive control, whereas in case of the negative control, sulfur element and other metabolic
products were absent. High corrosion rate of steel in de-aerated SRB medium containing high Fe+2
concentration was detected by Booth et al. (1967). The corrosion rate of N-80 steel in SRB containing
test medium was six times more than that in abiotic medium (Sorioglu et al., 1997). Also in the
presence of SRB, there is accumulation of sulfur-based compounds whereas in its absence, there is not
(Castaneda and Benetton, 2008). Li et al. (2009) reported that pitting corrosion of steels is a very
complex process in the media inoculated with SRB. On the other hand, there was no clear difference
in the SEM images and EDX analysis between the negative control coupon sample (Fig. 8-A) and
coupons samples treated at the lethal doses of nitrite, microwave, ultraviolet and gamma radiation as
all of them exhibited clear non corroded surfaces (Fig. 8- F, H, J, L) and the absence of sulfur element
demonstrating the absence of bacterial activity at such treatment doses. So they can de used efficiently
for controlling the corrosive effect of SRB. Hubert et al. (2005) observed absence of corrosion during
the 64 day treatment with 20 mM nitrite. Videla and Herrera (2009) reported that inorganic anions
such as nitrite form an ionically bonded surface compound which produces a barrier to corrosion
reaction.
469
Table (1): The EDX analysis of metal coupons surfaces different immersed at the modified medium inoculated with SRB of
different treatments.
Sample
Element (Wt %)- Total= 100

Fe
Cr
Al
Si
Mn
Ca
Cl
Mg
Negative control sample: (Without SRB)
98.5
0.07
0.34
0.25
0.65
0.07
0.07
Positive control sample: (With untreated SRB)
92.21
1.61
0.12
1.0
0.61
0.21
0.13
77.02
0.07
0.2
0.72
0.56
2.11
0.06
11.87
3.58
2.62
1.19
98.11
0.22
0.07
0.13
0.26
0.68
0.11
0.34
0.07
97.28
0.28
0.07
0.14
0.84
0.72
0.26
0.28
0.12
96.46
0.08
0.29
1.15
0.75
0.35
0.38
0.54
96.25
0.12
0.06
0.23
0.74
0.70
0.73
0.88
0.28
97.20
0.10
0.06
0.62
0.66
0.48
0.60
0.29
94.32
0.96
0.05
0.12
1.1
0.73
1.02
1.07
0.64
98.24
0.08
0.15
0.53
0.66
0.23
0.11
89.09
0.79
0.06
0.08
1.13
0.58
0.47
3.86
2.99
0.61
0.34
97.41
0.08
0.40
0.41
0.63
0.25
0.71
0.12
Treated samples
Campion A
Nitrate
Nitrite
Microwave
U.V.
Gamma radiation
Dose
Sublethal:(50 ppm)
(10 mM/l)
Sublethal: (4 mM/l)
Lethal: (6 mM/l)
Sublethal: (40 sec.)
Lethal: (50 sec.)
Sublethal: (3hours)
Lethal: (4 hours)
Sublethal:(0.7 KGy)
Lethal: (0.9 KGy)
470
(SEM images)
(EDX analysis charts)
A- Negative control sample (without SRB).
B- Positive control sample (with untreated SRB).
C- Champion-A biocide (50ppm).
471
Continued
D- Nitrate (10 mM/l).
E- Nitrite (4 mM/l).
F- Nitrite (6 mM/l).
472
Continued
G- Microwave (40 sec.)
H- Microwave (50 sec.).
I- Ultraviolet (3 hours).
J- Ultraviolet (4hours).
473
Continued
K- radiation (0.7KGy).
L- radiation (0.9KGy).
Fig. (50): Scanning electron microscopy (SEM) micrograph (Left column) and EDX analysis
(Right column) of metal coupons surfaces immersed at the modified medium:
A- without SRB, B- with untreated SRB, C- with SRB pretreated with Champion-A biocide
(50ppm), D- containing nitrate (10 mM/l) and inoculated with SRB, E- containing nitrite (4
mM/l) and inoculated with SRB, F- containing nitrite (6 mM/l) and inoculated with SRB, Gwith SRB pre-exposed to microwave for(40 sec.), H- with SRB pre-exposed to microwave
for(50 sec.), I- with SRB pre-exposed to ultraviolet for(3 hours), J- with SRB pre-exposed to
ultraviolet for (4 hours), K- with SRB pre-exposed to gamma radiation dose (0.7 KGy), L- with
SRB pre-exposed to gamma radiation dose (0.9 KGy).
Also Fig. (8) and Table (1) show a clear variation between the positive control sample and all
the treated samples at the sublethal doses of (nitrite, microwave, U.V. and gamma radiation), where
the pit density and amount of sulfur element precipitated on the metal surface of positive control
sample as shown in Fig. (50-B) were more intensive and much greater than other sublethal dose
treated samples (Fig. 8- E, G, I, K). The localized damage to the metallic surface of control sample and
the higher concentration of sulfur based compound and other potentially biotical generated corrosion
influencing compounds are likely to be enhanced due to SRB metabolism and biofilm formation
(Castaneda and Benetton, 2008).
474
The SEM images and EDX analysis presented at Fig. (8- B, D) and Table (1) show the presence
of pitting corrosion and sulfur element on the coupon surface of nitrate treated sample but still much
less than what was detected on coupon surface of the positive control sample. Bodtker et al. (2008)
found that long term nitrate treatment provided efficient inhibition of SRB activity at Gulfaks oil field,
the reduction in activity was followed by a significant reduction in corrosion up to 40%. Also the
corrosion rated were less than 1.5 mpy when coupons were incubated in medium containing sulfate
and nitrate than sulfate only. Further more the occurrence of pitting corrosion was fairly low under all
circumstances (Dunsmore et al., 2004). On the other hand some researchers have observed an
increase in corrosion rates related to nitrate mediated souring control (Nemati et al., 2001; Rempel et
al., 2006 and Schwermer et al., 2008).
It was observed that the applied biocide (Champion- A) of the dose (50 ppm) showed the worst
treatment as it exhibited an intense localized corrosion of the coupon surface although the EDX
analysis presented in Table (45) and Fig. (50-C) did not show any presence of sulfur indicating
complete suppression of SRB activity at such treatment. This corroded aspect of the coupon surface of
biocide treated sample can be attributed to the corrosive action of the used biocide itself or killing the
microbial community members that offer protection against corrosion. This was previously reported
by Zuo et al. (2004). Also the tendency of increasing corrosion rate with biocide treatment has also
been observed in water injection systems at the Veslefrikk and Gulfaks field (North Sea)
(Thorstenson et al., 2002 and Sunde et al., 2004). The mean corrosion rate observed at the well head
of a studied oil field during nitrate treatment was lower than observed during biocide treatment, but
the reduction was not significant (Bodtker et al., 2008). These results impose the use of other control
treatments rather than the traditional and common use of chemical biocides which resulted corrosion
aspects on the surfaces of the studied metal coupons.
REFERENCES
(1)
Alabostro, F.; Pineda, A.; Pangan, A. and Delvalle, M. (1987). Food preservation by irradiation.
Vol. 1, (IAEA, Viena), p. 282.
(2)
Ash, M. and Ash, D. (2001). Handbook of corrosion inhibitors. NACE International, Houston.
(3)
ASTM G 1-72 (1993). Standard recommended practice for preparing, cleaning and evaluating
corrosion test specimens. Annual Book of ASTM standards. American Society for Testing and
Materials, Philadelphia, vol. 03.02, p. 33.
(4)
ASTM, D 4412-84 (1990). Standard test methods for sulfatereducing bacteria in water and
water formed deposits. Annual Book of ASTM standards, American Society for Testing and
Materials, vol. 11.01.
(5)
Beech I.B., and Gaylarde C.C Recent advances in the study of biocorrosion - an
.Rev.Microbiol 30, 177 (1999).
(6)
Bjorndalen, N.; Mustafiz, S.; Tango, M. and Islam, M. (2003), A novel technique for prevention
of microbial corrosion. Energy Sources, 25, p. 945.
(7)
Bodtker, G.; Thorstenson, T.; Lillebo, B.; Thorbjornsen, B.; Ulvoen, H.; Sunde, E. and Torsvik,
T. (2008). The effect of long term nitrate treatment on SRB activity, corrosion rate and bacterial
community composition in offshore water injection systems. J. Ind. Microbiol. Biotechnol., 35,
p. 1625.
(8)
Booth, G. (1964). Sulphur bacteria in relation to corrosion. J. Appl. Bacteriol., 27, p. 174.
475
overview
(9)
Booth, G.H.; Robb, J.A. and Wakerly, D.S. (1967).

Congress of Metallic Corrosion, Moscow, 2, p. 542.
In: Proceedings of 3rd International
(10) Castaneda, H. and Benetton, X., (2008). SRB-biofilm influence in active corrosion sites formed
at the steel electrolyte interface when exposed to artificial seawater conditions. Corrosion
Science, 50, p. 1169.
(11) Cirne, D.; Van der Zee, F.; Fernandez- Polanco, M. and Fernandez- Polanco, F. (2008). Control
of supplied during anaerobic treatment of S- containing wastewaters by adding limited amounts
of oxygen or nitrate, Rev. Environ. Sci. Biotechnol., 7, p. 93.
(12) Cochrane, W. (1950). Estimation of bacterial densities by means of the most probable number.
Biometrics, 6, p. 105.
(13) Cord- Ruwish, R. (1985). A quick method for the determination of dissolved and precipitated
sulfides in cultures of sulfatereducing bacteria. J. Microbiol. Methods, 4, p. 33.
(14) Davidova, Hicks M.S., Fedorak P.M., and Suflita J.M. The influence of nitrate on microbial
processes in oil industry production waters J. Ind. Microbiol. Biotechnol. 27 ,80 (2001).
(15) Dunsmore, B.; Whitfield, T.; Lawson, P. and Collins, M. (2004). Corrosion by sulfatereducing bacteria that utilize nitrate. Paper No. 04763. CORROSION 2004, NACE
International, USA.
(16) Eckardt-Schupp, F. and Klause, C. (1999). Radiation inducible DNA repair processes in
eukaryotes. Bioch., 81, p. 161.
(17) El-Batal, A. and Khalaf, M. (2003). Wheat Bran as a substrate for enhanced thermostable alphaamulase production by gamma irradiated Bacillus megaterium in solid state fermentation. Egypt.
J. Rad. Sci. Applic., 16, 443.
(18) Fujikawa, H. and Ohta, K. (1994). Patterns of bacterial destruction in solutions by microwave
irradiation. Journal of Applied Bacteriology, 67, p. 389.
(19) Garcia De-lomas, J.; Corzo, A.; Carmen portillo, M.; Gonzalez, M.; Andrades, J.; Saiz-Jimenez,
C. and Garcia-Robledo, E. (2007). Nitrate stimulation of indigenous nitrate reducing, sulfide
oxidizing bacterial community in waste water. anaerobic biofilms, 41, p.3121.
(20) Gardner, L. and Stewart, P. (2002). Action of glutaraldehyde and nitrite against sulfate reducing
bacterial biofils. J. Ind. Microbiol. Biotechnol., 29, p. 354.
(21) Gonzalez- Rodriguez, C.; Rodriguez- Gomez, J. and Genesca- LIongueras, J. (2008). The
influence of Desulfovibrio vulgaris on the efficiency of imidazoline as a corrosion inhibitor on
low- carbon steel in seawater.Elecrochemica Acta, 54, p. 86.
(22) Hamid, M.; Thomas, T.; El- Saba, A.; Stapleton, W.; Sakla, A.; Rahman, A.; Byrne, P.;
VanLanding- ham, D. and McCombs, C. (2001). The effects of microwaves on airborne
microorganisms. Journal of Microwave Power and Electromagnetic Energy, 36, p. 37.
(23) Heitz, E. (1996). Microbially Influenced Corrosion of Materials, Springer-Verlag, Berlin and
Heidelberg, Gmbh Co., Chapter VIII, p. 105.
(24) Hubbert C., Nemati M., Jenneman G., and Voordow G. Corrosion risk associated with
microbial souring control using nitrate or nitrite Appl. Microbiol. Biotechnol. 68, 272 (2005).
476
(25) Hubert, C.; Nemati, M.; Jenneman, G. and Voordouw, G. (2003). Containment of biogenic
sulfide production in continuous up- flow packed- bed bioreactors with nitrate or nitrite.
Biotechnol. Prog., 19, p.338.
(26) Javaherdashti R. Microbiologically Influenced Corrosion (MIC) Engineering materials and
processes, microbiologically influenced corrosion Hand book, chapter 4 (2008).
(27) Jenneman, G.; McInerney, M. and Knapp, R. (1986). Effect of nitrate on biogenic sulfide
production. Appl. Environ. Microbiol., 51, p. 1205.
(28) Kjellerup, B.; Veeh, R.; Sumithraratne, P.; Thomsen, T.; Buckingham- Meyer, K.; Frolunde, B.
and Sturman, P. (2005). Monitoring of microbial souring in chemically treated, produced water
biofilm systems using molecular techniques. J. Ind. Microbiol. Biotechnol., 32, p.163.
(29) Li, F.; An, M.; Liu, G. and Duan, D. (2009). Effects of sulfidation of passive film in the
presence of SRB on the pitting corrosion behaviors of stainless steels. Material Chemistry and
Physics, 113, p. 971.
(30) Magot M., Ollivier B., and Patel B. Microbiology of petroleum reservoirs Antoni van
Leeuwenhoek. 77 ,103 (2000).
(31) Meleigy, S. (2009). Effect of cultural conditions and gamma irradiation on the production of galactosidase enzyme from Kluyveromyces maraxianus EMCC75 grown in milk permeate. Arab
J. Nuclear Science and Applications, 42, (3).
(32) Miranda,E. , M. Bethencourt, F.J. Botana, M.J. Cano, J.M. Sanchez-Amaya, A. Corzo, J.
Garca de Lomas, M.L. Fardeau and B. Ollivier Biocorrosion of carbon steel alloys by a
hydrogenotrophic sulfate-reducing bacterium Desulfovibrio capillatus isolated from a Mexican
oilfield separator Corrosion Science 48 2417 (2006).
(33) Mittelman, M. (1990). Bacterial growth and biofouling and biocorrosion. Stuttgart, September
13-14 1990 (Flemming H-C, Geesey GG, eds) Springer- Verlag Berlin, Heidelberg.
(34) Mohanakrishnan, J.; Gutierrez, O.; Meyer, R. and Yuan, Z. (2008). Nitrite effectively inhibits
sulfide and methane production in a laboratory scale sewer reactor. Water Research, 42, p. 3961.
(35) Myhr, S.; Lillebo, B.; Sunde, E. and Beeder, J. (2002). Inhibition of microbial H2S production in
oil reservoir model column by nitrate injection. Appl. Microbiol. Biotechnol., 58, p. 400.
(36) Nemati M., Jenneman G., and Voordow G. Mechanistic study of microbial control of hydrogen
sulfide production in oil reservoirs Biotechnol. Bioeng. 74, 424 (2001).
(37) Rempel, C.; Evitts, R. and Nemati, M. (2006). Dynamics of corrosion rates associated with
nitrite or nitrate mediated control of souring under biological conditions simulating an oil
reservoir. J. Ind. Microbiol. Biotechnol., 33, p. 878.
(38) Saiz- Jimenez, C. (2001). The biodeterioration of building materials In: A practical manual on
microbiologically influenced corrosion (Stoecket II JG, eds) 2nd edn, NACE International 2001.
(39) Sato, S.; Shibata, C. and Yazu, M. (1996). Nonthermal killing effect of microwave irradiation.
Journal of Appl. Bacteriol., 10, p. 145.
(40) Schwermer, U.; Lavik, G.; Abed, R.; Dunsmore, B.; Ferdelman, T.; Stoodley, P.; Gieseke, A.
and de Beer, D. (2008). Impact of nitrate on the structure and function of bacterial biofilm
communities in pipelines used for injection of seawater into oil fields. Appl. Environ.
Microbiol., 74, p. 2841.
477
(41) Sorioglu, F.; Javaherdashti, R. and Aksoz, N. (1997). Corrosion of a drilling pipe steel in an
environment containing sulphate- reducing bacteria. Int. J. Pres. Ves. & Piping, 73, p. 127.
(42) Sturman, P. and Goeres, D. (1999). Control of hydrogen sulfide in oil and gas wells with nitrite
injection. SPE Annu. Tech. Conf. Exhib.,p. 357.
(43) Sunde, E.; Lilebo, B.; Bodtker, G.; Torsvik, T., and Thorstenson, T. (2004). H2S inhibition by
nitrate injection to the Gulfaks field, paper no. 04760 Proceedings of Corrosion/2004. NACE
International, Houston, TX.
(44) Thorstenson, T.; Boedtker, G.; Sunde, E. and Beeder, J. (2002). Biocide replacement by nitrate
in sea water injection systems, Corrosion, 33, p. 1.
(45) Videla, H. and Herrera, L. (2009). Understanding microbial inhibition of corrosion. A
comperehensive overview. International Biodeter. Biodegrad., 63, p. 896.
(46) Von- Reg, H. and Sand, W. (1998). Evaluation of biocide fficacy by microcalorimtric
determination of microbial activity in biofilms. J. Microbiol. Methods, 33, p. 227.
(47) Wargin, A.; Olanczuk- Neyman, K. and Skucha, M. (2007). Sulphate- reducing bacteria, Their
properties and methods of elimination from groundwater. Polish Journal of Environmental
Studies, 16, pp. 639.
(48) Zhang, L.; De Schryver, P.; De Gusseme, B.; De Muynck, W.; Boon, N. and Verstraete, W.
(2008). Chemical and biological technologies for hydrogen sulfide emission control in sewer
systems: A review. Water Research, 42, p. 1.
(49) Zuo, R.; Ornek, D.; Syrett, B.; Green, R.; Hsu, C.; Mansfeld, F. and Wood, T. (2004). Inhibiting
mild steel corrosion from sulfatereducing bacteria using antimicrobial- producing biofilms in
Three- Mile Island process water. Appl. Microbiol. Biotechnol., 64, p. 275.
478

Control of Microbiologically Induced Corrosion in

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Control of Microbiologically Induced Corrosion in

Uploaded by

Copyright:

Available Formats

Arab Journal of Nuclear Sciences and Applications, 45(2)460-478(2012)