Chinese J. Chem. Eng.

, 14(6) 829834 (2006)

RESEARCH NOTES

Corrosion and Electrochemical Behavior of 316L Stainless Steel in

Sulfate-reducing and Iron-oxidizing Bacteria Solutions*
XU Congmin()a, ZHANG Yaoheng()a, CHENG Guangxu()a,** and
ZHU Wensheng()b
a

Department of Chemical Engineering, Xian Jiaotong University, Xian 710049, China

Research and Technology Center of Lanzhou Oil Refinery Factory, PetroChina Company Ltd., Lanzhou 730060,
China
b

Abstract Corrosion and electrochemical behavior of 316L stainless steel was investigated in the presence of
aerobic iron-oxidizing bacteria (IOB) and anaerobic sulfate-reducing bacteria (SRB) isolated from cooling water
systems in an oil refinery using electrochemical measurement, scanning electron microscopy (SEM) and energy
dispersive atom X-ray analysis(EDAX). The results show the corrosion potential and pitting potential of 316L
stainless steel decrease distinctly in the presence of bacteria, in comparison with those observed in sterile medium
under the same exposure time. SEM morphologies have shown that 316L stainless steel reveals no signs of pitting
attack in the sterile medium. However, micrometer-scale corrosion pits were observed on 316L stainless steel surface in the presence of bacteria. The presence of SRB leads to higher corrosion rates than IOB. The interactions
between the stainless steel surface, abiotic corrosion products, and bacterial cells and their metabolic products increased the corrosion damage degree of the passive film and accelerated pitting propagation.
Keywords sulfate-reducing bacteria (SRB), iron-oxidizing bacteria (IOB), 316L stainless steel, pitting corrosion,
electrochemical behavior

INTRODUCTION
Type 316L stainless steel has good corrosion resistance and has been used increasingly for cooling
water service in the chemical, petrochemical and
power utility industries. However stainless steel is
susceptible to localized corrosion by chloride ions and
reduced sulfur compounds[1]. The presence of microorganisms on a metal surface often leads to highly
localized damages in the concentration of the electrolyte constituents, pH and oxygen levels[2]. These microorganisms and their metabolic activity have influenced severely the corrosion process, and often
stimulated localized forms of corrosion[3]. During the
last several years, various cases of corrosion damage
caused by bacteria were observed in cooling water
system of oil refinery, sulfate-reducing bacteria (SRB)
and iron-oxidizing bacteria (IOB) were the most troublesome group of bacteria on tubercular corrosion and
induced microbiologically influenced corrosion in
cooling circuit, caused poor water quality and equipment clogging, pipe punctures and high corrosion
rates, resulted in the serious pitting corrosion of carbon steel equipment[47]. The biological corrosion of
steels has received increasing consideration in the last
few decades. Similar studies by Duan et al.[8] showed
the characteristics of sulfide corrosion products on
316L stainless steel surfaces with the presence of SRB
in seawater and soil environment, the study of Romero
et al.[9] indicated that attack morphology of carbon
steel generated by SRB in systems for secondary re-

covery of crude oil is characterized by rounded holes

in chains or groups, Chamritski et al.[10] founded that
IOB could give rise to minor localized corrosion
damage of 304L stainless steel by under-deposit (i.e.,
crivice) corrosion in natural spring water. However,
few studies were reported about the corrosion and
electrochemical behavior of stainless steel in cooling
water system of oil refinery.
The present study is aimed to gain better understanding of corrosion process characteristics and electrochemical behavior of 316L stainless steel in the
media of anaerobic SRB and aerobic IOB separated
from cooling water systems of oil refinery using open
circuit potential measurement, electrochemical measurement, scanning electron microscopy (SEM) and
energy dispersive atomi X-ray analysis (EDAX) techniques.
2 EXPERIMENTAL
2.1 Preparation of specimen
The corrosion specimens were cut from type
316L stainless steel sheet, the nominal elemental
composition (%, by mass) of 316L SS specimens was:
C 0.029, Cr 16.97, Ni 10.11, Mo 2.04, Mn 1.38, Si
0.39, P 0.031, S 0.005. Disc shape specimens with a
diameter of 18mm and thickness of 2mm were used
for electrochemical measurements, rectangular specimens with dimensions 30mm25mm2mm were
used for biofilm observation. To create working electrodes, an electrical contact to each sample was

Received 2006-03-20, accepted 2006-09-18.

* Supported by the National Natural Science Foundation of China (No.20576108).
** To whom correspondence should be addressed. E-mail: gxcheng@mail.xjtu.edu.cn

Chinese J. Ch. E. (Vol. 14, No.6)

830
Table 1

Analytical results for cooling water sampled from oil refinery (mgL1)

Cl

CO32

HCO3

Ca2+

Mg2+

SO 24

pH

Total hardness

Total dissolved solid

Salt

242.66

19.246

435.504

112.30

200.15

240.24

8.16

1105.35

932

0.9

provided by a long copper wire connected to the back

of each specimen mounted in an epoxy resin, then the
specimens were abraded through 240, 400 and
600-grit silicon carbide metallurgical paper, degreased
in acetone, washed with sterile distilled water and
dried in a desiccator until use.
2.2

Microbiological cultivation and inoculation

Experimental SRB and IOB were isolated from
cyclic the cooling water system of an oil refinery plant,
the chemical composition of cooling water is provided
in Table 1. IOB species isolated from cyclic cooling
water system was identified as Leptothrix sp. This is
the most common iron storing ensheathed bacterium
apparently occurring in slow running, ferrous
iron-containing waters, poor in decomposable organic
material. Leptothrix species oxidize ferrous ions to
ferric ions to obtain their energy; SRB identified was
Desulfovibrio sp. This is documented to show aggressive corrosion with many metals under anaerobic conditions. SRB and IOB were cultivated separately in
appropriate media. SRB culture was grown in corrected

PostgateC medium: 0.5gL 1 KH2PO4, 1.0gL 1

1
1
Na2SO4, 0.06gL
CaCl22H2O,
NH4Cl, 4.5gL

0.06gL 1 MgSO47H2O, 6gL 1 sodium lactate,

1.0gL 1 Yeast extract, 0.004gL 1 FeSO47H2O,

1
0.3gL sodium citratepH 7.2under anaerobic
conditions. IOB culture was grown in Winogradski

nutrient medium: 0.5gL 1 K2HPO4, 0.5gL 1 NaNO3,

1
1

CaCl2, 0.5gL
MgSO47H2O, 0.5gL 1
0.2gL
1
NH4NO3, 6.0gL ammonium iron citrate (pH 6.8)
under aerobic chamber. These solutions were autoclaved at 121 for 20min. Enriched cultures was
incubated at 30. Enriched cultures were used as the
corrosion cell inoculum. Test cells were inoculated

with 5% (by volume) at approximately 108cellsml 1

of each of the selected cultures.
Electrochemical measurements
All electrochemical tests were carried out in a 2L
corrosion cell, with a three-electrode system, the
measurements were done with M263A potentiostat
and phase lock-in amplifier (EG & G, USA). Working
electrode potentials were referred to a saturated calomel electrode (SCE). The counter electrode was a
Pt-plate. Polarization curves were determined poten
tiodynamically with a scan rate of 0.5mVs 1. EIS
measurements were made at the open circuit potential
using a 10mV amplitude sinusoidal signal over frequencies ranging from 5mHz to 100kHz. All measurements were carried out at 30 for optimum bacteria growth.

2.4

Surface analysis
The test coupons were examined for surface
biofilm and corrosion features using SEM and EDAX.
The coupons with biofilm were immersed for 15min
in a 4% glutaraldehyde solution in order to fix the
biofilm to the stainless steel surface, and then become
dehydrated using four ethanol solutions (15min each):
25%, 50%, 75% and 100% successively. After that,
the samples were taken to the SEM and EDAX for
their surface analysis.
3 RESULTS AND DISCUSSION
3.1 Corrosion potential vs. time
Each test was allowed to run until the corrosion
potential (Ecorr) and the polarization resistance(Rp)
reached their asymptotic values. Fig.1 shows the
variations of the corrosion potential (Ecorr) with the
immersion time for stainless steel in sterile medium,
IOB and SRB solutions at 30. In the sterile medium,
no significant changes in Ecorr occur, indicating that
the specimen was in a passive state during the whole
test session. Ecorr of both electrodes were reduced by
about 0.36V (from 0.06 to 0.42V vs. SCE)
and 0.46V (from 0.06 to 0.52V vs. SCE) in
IOB solution and SRB solution respectively. Moreover SRB make Ecorr values drop at a rate faster than
that observed in the presence of IOB. After about 10d
of immersion, Ecorr of both electrodes becomes stabilized. The decrease of Ecorr was related to the dissolution of electrode surface passive film induced by
metabolic activity of SRB and IOB during the immersion.

2.3

December, 2006

Figure 1

3.2

Variation of corrosion potential with time for

stainless steel in different solutions
sterile medium; IOB; SRB

Electrochemical test
Figure 2 illustrates the potentiodynamic polarization curves of stainless steel electrodes in sterile medium, IOB and SRB solutions after 4d immersion at
30. The width of the passive range in sterile medium is the largest, that is 1.40V (0.15 to 1.25 VSCE),
and the value of pitting potential (Epit) is the highest,

Corrosion and Electrochemical Behavior of 316L Stainless Steel in SRB and IOB Solutions

about +1.25V, which shows a good pitting resistance.

Extended passive region, from 0.3 to +0.48V, is
found for stainless steel in IOB solution, and unstable
pitting was initiated at 0.48V. A second passivation
occurred at 0.52V, but the passive region was very
narrow. At around 0.82V, stable pitting was developed.
In contrast, a narrower passive region (from 0.53 to
0.29V) in SRB solution was observed before unstable pitting potential was reached, indicating decrease
in passive state stability. At potentials around 0.88V,
stable pitting corrosion occurred. There is a tendency
for polarization curves to shift right (to higher current densities) in sterile medium, IOB and SRB solutions respectively. In addition, Fig.2 also show that the
anodic polarization current density increases significantly and that the width of the passive range of
potentials and corrosion potential decrease in sterile

831

medium, IOB and SRB solutions in turn, indicating

the corrosion rate (corrosion current density) was the
highest in SRB solution, higher in IOB solution and
the lowest in sterile medium.
Figure 3 presents the cyclic polarization curves
for stainless steel electrodes in sterile medium, SRB
and IOB solutions after 20d immersion at 30 with a

scan rate of 0.5mVs 1. The potential where the loop

was closed is referred to as the repassivated potential
(Erp). Pits grow between Epit and Erp, but the nucleation of new pits only take place above Epit. Below Erp
pit growth is not observed. The local environment
chemistry affects both Epit and Erp[11]. Therefore, the
presence of microorganisms can significantly affect
both potentials. As can be seen from Fig. 3, during the
reverse scan period in sterile medium, smaller current
densities are recorded for the same values of potential

Figure 2

Anodic polarization curves for stainless steel in three different solutions after 4d of immersion at 30
1sterile medium; 2SRB; 3IOB

Figure 3

Cyclic polarization curves for stainless steel in three different solutions after 20d of immersion at 30
1sterile medium; 2SRB; 3IOB
Chinese J. Ch. E. 14(6) 829 (2006)

Chinese J. Ch. E. (Vol. 14, No.6)

832

and the loop has a small area. The final Ecorr at +0.88V
is much more noble than the starting Ecorr at 0.05V,
indicating stainless steel specimen is efficiently passivated from the moment of its immersion. In two biological solutions, following passivity breakdown and
reversing the potential direction, a pronounced hysteresis was observed. In the presence of IOB, pitting
corrosion was initiated at 0.8V(positive scan) and Erp
at 0.1V. In SRB solution, Epit and Erp were much
lower than that caused by IOB, which is at a potential
of 0.45V and 0.08V respectively. Compared with
Fig.2, Epit values of two electrodes in IOB and SRB
solutions decreased with increasing exposure time,
which shows the pitting corrosion of stainless steel
was further enhanced with exposure time.
As a complementary technique, electrochemical
impedance spectrum (EIS) was conducted to confirm
the trends of the corrosion rates determined by potentiodynamic polarization method. Nyquist diagrams of
stainless steel in sterile medium, SRB solution and
IOB solution are given in Fig.4. The Nyquist plot in
sterile medium shows an open capacitance arc. It is
evident that in the frequency range of the measurement, two diagrams in two biological solutions reveal
qualitatively similar features. The effect of SRB is to
decrease the magnitude of the impedance value compared to that of IOB. The impedance induced by SRB
is the smallest, followed by IOB and sterile medium.

Figure 4

Niquist plots for 316L SS in two biological solutions

sterile medium (exp.); IOB (exp.); SRB (exp.);
fitted values

3.3

Surface analysis
SEM was carried to validate the adhesion of the
microorganisms to the stainless steel surface and also
to analysis microbial diversity. Fig.5 shows a detail of
biofilm developed on stainless steel surface exposed
to SRB solution and IOB solution for 15d. As seen in
Fig.5(a), there was a predominance of rod-shaped
SRB cells in the presence of SRB, where a high cell
density with the typical morphology of the genus used
(Desulfovibrio between 1.5m to 2.0m) can be observed. In the presence of IOB, there was a dominance
of spherical IOB cells (Leptothrix between 1.0m to
1.5m), where big colonies were observed [Fig.5(b)].
To analyze the effects of corrosion product layer,
December, 2006

(a) SRB

Figure 5

(b) IOB
SEM micrograph of biofilms on electrode
surface after 15d of exposure

SEM coupled with EDAX analysis patterns were

taken to identify the constituents of corrosion product
layer formation on stainless steel surface. SEM and
EDAX analysis on the samples after 15d of exposure
in SRB solution and IOB solution are shown in Figs.6
and 7. As can be seen from Fig.6(a), specimens were
covered with black corrosion products which were
dense, brittle and lumpy deposits, the cracks eventually were clearly seen on the surface, the characteristic
color of iron sulfide was observed in the cell with
SRB, produced by the reaction of the sulfides generated by the SRB with the ferrous salts present in the
culture medium[12]. High sulfur content and small isolated heaps were observed in Fig.6(b). It is important
to point out that, in accordance with the pH of medium (7.2), this compound could be kansite (Fe9S8),
which has very poor protective properties[13]. In the
presence of IOB, the corrosion products were porous
brownie deposits [Fig.7(a)], EDAX analysis showed
that these deposits evidently are iron oxides.
Typical SEM micrographs of corrosion pits for
stainless steel coupons are presented in Fig.8 for 30d.
The SEM analysis on the stainless steel surface indicated that the metal exposed to sterile medium, exhibited basically no localized corrosion observed on the
metal surface under open circuit conditions for 30d
[Fig.8(a)]. Once the corrosion products film was
removed from the steel exposed to two biological
solutions, localized corrosion was observed on the
metal surface, as shown in Figs.8(b) and 8(c). In SRB

Corrosion and Electrochemical Behavior of 316L Stainless Steel in SRB and IOB Solutions

(a) Sterile medium

(a) SEM

(b) EDAX (spectrum 1)

Figure 6 SEM micrographs and EDAX analysis of
corrosion products induced by SRB on sample
surface after 15d of exposure

(a) SEM

(b) EDAX (spectrum 1)

Figure 7 SEM micrographs and EDAX analysis of
corrosion products induced by IOB on sample
surface after 15d of exposure

solution, the presence of SRB apparently initiates pitting corrosion of the exposed specimens as indicated
by the presence of large and deep corrosion pits as
shown in Fig.8(b). As SRB have been documented to
show aggressive corrosion with many metals under

833

(b) SRB solution

Figure 8

(c) IOB solution

SEM micrographs of corrosion pits on sample
surface after 30d of exposure

anaerobic conditions. SRB are ubiquitous and easy to

culture, they produce hydrogen sulfide that can react
with stainless steel in sulfate-containing environments.
Sulfide produced by the SRB then migrates to edges
of the deposits where it is oxidized to thiosulfate,
which is a well-known activator of pitting corrosion[14].
The estimated contribution of SRB to the total bacterial biomass of the systems always exceeded that of
the IOB consortium, which was consistently the lowest[15]. In IOB solution, the corrosion damage observed on coupons surface illustrates only a minor and
shallow circular corrosion pits [Fig.8(c)]. This is attributed to iron bacterias slow metabolic ability to
oxidize ferrous ions to ferric and then form a low density hydrated iron oxide in the corrosion tubercles.
The metabolic ability is key factor for corrosion of
steel, as the metabolism of these microorganisms is
Chinese J. Ch. E. 14(6) 829 (2006)

Chinese J. Ch. E. (Vol. 14, No.6)

834

very slow, especially under non-optimum, circumneutral pH (7.2) conditions[15]. The results indicate that
pitting corrosions of 316L SS in SRB solution is the
most severe, followed by IOB and sterile medium.
This is consistent with the result of the electrochemical measurement.

7
8

NOMENCLATURE
Ecorr
Epit
Erp
Z
Z

corrosion potential, V
pitting potential, V
repassivated potential, V
real resistance impedance, cm2
imaginative resistance impedance, cm2

10

REFERENCES
1

2
3

4
5
6

Ismail, Kh.M., Jayaraman, A., Wood, T.K., Earthman,

J.C., The influence of bacteria on the passive film stability of 304 stainless steel, Electrochim. Acta, 44,
46854692(1999).
Costerton, J.W., Lewandowski, Z., Caldwell, D.E.,
Korber, D.R., Lappin-Scott, H.M., Microbial biofilms,
Annual Review of Microbiology, 49, 711(1995).
Dexter, S.C., Duquette, D.J., Siebert, O.W., Videla, H.A.,
Use and limitations of electrochemical techniques for
investigating microbiological corrosion, Corrosion,
47(4), 308318(1991).
Rosenfeld, I.L., Corrosion Inhibitors, Chemistry, Publishing Ltd., Moscow (1977).
Starosvetsky, J.O., Armon, R., Groysman, Fouling of
carbon steel heat exchanger caused by iron bacteria,
Mater. Performance, 38 (1), 5561(1999).
Rao, T.S., Sairam, T.N., Viswanathan, B., Nair, K.V.K.,

December, 2006

11
12

13
14
15

Carbon steel corrosion by iron oxidising and sulphate

reducing bacteria in a freshwater cooling system, Corros. Sci., 42, 14171431(2000).
Starosvetsky, D., Armon, R., Pitting corrosion of carbon
steel caused by iron bacteria, Int. Biodeter. Biodegr., 47,
7987(2001).
Duan, J.Z., Hou, B.R., Yu, Z.G., Characteristics of sulfide corrosion products on 316L stainless steel surfaces
in the presence of sulfate-reducing bacteria, Mat. Sci.
Eng. C, 26, 624629(2006).
de Romero, M., Duque, Z., Rodrguez, L., de Rincn, O.,
Araujo, I., A study of microbiologically induced corrosion by sulfate-reducing bacteria on carbon steel using
hydrogen permeation, Corrosion, 61, 6874(2005).
Chamritski, I.G., Burns, G.R., Webster, B.J., Effect of
iron-oxidizing bacteria on pitting of stainless steel,
Corrosion, 60(7), 658669(2004).
Khan, M.A., Williams, R.L., Williams, D.F., Conjoint
corrosion and wear in titanium alloys, Biomaterials, 20,
765772(1999).
de Mele, M.F.L., Moreno, D.A., Ibars, J.R., Videla, H.A.,
Effect of inorganic and biogenic sulfide on localized
corrosion of heat-treated type 304 stainless steel, Corrosion, 47, 2430(1991).
Scott, P.J.B., Michael, D., Survey of field kits for sulfate-reducing bacteria, Mater. Performance, 31, 6468
(1992).
Felder, C.M., Stein, A.A., Microbiologically influenced
corrosion of stainless steel weld and base metal-four-year
field test results, Corrosion, 50, 275(1994).
Pitonzo, B.J., Castro, P., Amy, P.S., Southam, G., Jones,
D.A., Ringelbery, D., Microbiologically influenced
corrosion capability of bacteria isolated from yucca
mountain, Corrosion, 60, 6474(2004).