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Corrosion Science 65 (2012) 113118

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Evaluation of microbiologically inuenced corrosion inhibition on NiCo alloy

coatings by Aeromonas salmonicida and Clavibacter michiganensis
Nalan Oya San a, Hasan Nazr b, Gnl Dnmez c,
Ankara University, Institute of Biotechnology, 06100 Besevler-Ankara, Turkey
Faculty of Science, Department of Chemistry, 06100 Besevler-Ankara, Turkey
Faculty of Science, Department of Biology, 06100 Besevler-Ankara, Turkey

a r t i c l e i n f o a b s t r a c t

Article history: Microorganisms are able to intensely change the electrochemical conditions at the metal/solution inter-
Received 15 February 2012 face by forming biolms. These changes can result in acceleration of corrosion to corrosion inhibition. In
Accepted 2 August 2012 this study microbiologically inuenced corrosion inhibition (MICI) has been observed for NiCo alloy
Available online 11 August 2012
coatings when two bacteria inoculated to the medium. Experiments were performed in sterile medium
and using Aeromonas salmonicida and Clavibacter michiganensis bacteria. After the inoculation of the bac-
Keywords: teria, a decrease detected at the Ecorr and Icorr values. Highest inhibition efciency (gp and gi) values were
A. Alloy
detected for C. michiganensis, as it caused a slower corrosion reaction.
A. Nickel
2012 Elsevier Ltd. All rights reserved.
C. Microbiological corrosion

1. Introduction tions have led to a new approach of corrosion protection based on

corrosion control using natural bacteria.
Microbiologically inuenced corrosion (MIC) has been dened The electrochemical properties of nickel coatings are well
as an electrochemical process in which the participation of micro- known. Their wide application is a result of specic properties of
organisms is able to initiate, facilitate, or accelerate the corrosion nickel, which exhibits good corrosion resistance in aggressive solu-
reaction [14]. The formation of biolms on materials immersed tions and a high catalytic activity for many electrochemical pro-
in an aquatic environment is a combined action by adhesion of or- cesses. In order to enhance the utilization of these materials,
ganic/inorganic macromolecules, exo polymeric substance (EPS) various modications can be applied, such as the use of nickel al-
production, microbial growth and hydrodynamic erosion. EPS are loys [1215]. In addition, electrodeposited NiCo alloy coatings
known to modify the substratum surface properties by modifying have high strength and hardness, specic magnetic properties,
surface charge, wettability and surface free energy resulting in en- good chemical stability, wear and corrosion-resistance [1618].
hanced corrosion or corrosion inhibition [5,6]. Two examples of bacteria isolated from the water sample of the
The general concept of MIC is based on the assumption that corroded water treatment system are; Aeromonas salmonicida and
microorganisms accelerate the kinetics of electrochemical reac- Clavibacter michiganensis. The former is A. salmonicida, Gram nega-
tions thereby producing increased corrosion rates [7,8]. Although tive, facultative, aquatic bacterium and caused the lethal disease in
Stadler et al. [9] demonstrated a corrosion protection of three dif- sh [19,20]. The latter is C. michiganensis, Gram positive, aerobic
ferent kinds of metal substrate in the presence of Sulfate-reducing and plant-pathogenic bacterium [21,22].
bacteria (SRB) and Pseudomonas strain, it is surprising that the pos- The major aim of the present study was to elucidate the inuence
sibility of corrosion inhibition caused by bacteria has rarely been of two bacterias corrosion inhibition on alloy coatings. Electro-
considered. Jayaraman et al. [10,11] have demonstrated that pro- chemical polarization and electrochemical impedance spectroscopy
tective biolms decreased the corrosion rate of mild steel, appar- (EIS) was used to characterize the corrosion inhibition process. The
ently by reducing the oxygen concentration at the metal surface. mass loss was determined by using the quartz crystal microbalance
Due to the high costs associated with repairing or replacing com- (QCM) method. In addition, scanning electron microscopy (SEM)
ponent damaged by MIC, the issue has become a signicant area was used to examine the electrode surface and energy dispersive
of scientic research. These results suggest that MICI is a more X-ray spectra (EDS) analysis was used to obtain elemental informa-
common phenomenon than was previously assumed and observa- tion from the corrosion products of some typical areas from the
surface of the alloy.
Corresponding author. Tel.: +90 312 212 67 20; fax: +90 312 223 23 99. Considering the lack of literature on the subject, this is the rst
E-mail address: (G. Dnmez).
report on investigation of microbiologically inuenced corrosion

0010-938X/$ - see front matter 2012 Elsevier Ltd. All rights reserved.
114 N.O. San et al. / Corrosion Science 65 (2012) 113118

inhibition of two novel bacteria on NiCo alloy coatings. It is (CHI125A). Surface area of a gold crystal was 0.204 cm2 and its
becoming apparent that MICI is a more common phenomenon than nominal resonant frequency was 7.995 MHz. Moreover, ST-42 car-
has been assumed previously. bon steel with a surface area of 0.204 cm2 was also used as a work-
ing electrode in other electrochemical studies. The Ag/AgCl (sat.
2. Material and methods KCl) electrode (CHI111) was used as the reference electrode and
a platinum wire (CHI115) was used as counter electrode for all
Two bacteria were isolated from water samples which were experiments. NiCo alloy was electrodeposited on the QCM and
taken from corroded water treatment system. Samples were incu- ST-42 electrodes from a nickelcopper bath containing NiSO47H20
bated in Nutrient Broth medium for enrichment, and subsequently (104 g/L), NiCl26H2O (3.5 g/L), H3BO3 (6.95 g/L) and CoSO47H2O
puried in sterile agar plates. The isolated bacteria were character- (5 g/L). Electrodeposition was performed chronoamperometrically
ized using of 16S rRNA nucleotide sequences and submitted to with 1.2 V (vs. Ag/AgCl) applied to the electrolysis system for
NCBI GenBank under Accession numbers GU907676 (A. salmonici- 150 s during the NiCo depositions. The Ni and Co contents of
da) and, GQ466171 (C. michiganensis) [23]. the deposit were also veried with an atomic absorption spectro-
The bacteria were cultured on a rotary shaker (New Brunswick photometer. So, the yield of coating for bath content was assumed
Scientic Innova 4230) at 30 1 C and 100 rpm in a N. Broth med- as 100%, where NiCo ratios were calculated as 90 and 10, respec-
ium used for enrichment culture, which contained 5.0 g peptone tively. Using the bathing solution explained, we electroplated the
from meat, 3.0 g meat extract, and 6.0 g sodium chloride in 1 L NiCo thin lm with a thickness of approximately 100 nm on
deionized water. The medium was autoclaved at 121 C for one side of the quartz crystal electrodes. Data acquisition software
15 min (Alp CL-40M, Japan). After 24 hours of incubation, cultures was used to record the mass change. Frequency change, Df, was
were centrifuged at 5000g, 10 min (Hettich EBA 12), and the bio- converted into the mass variation, Dm, using the Sauerbrey equa-
mass was used for bio-corrosion experiments. In order to ensure tion [25].
the reproducibility of the experiments, exponential phase bacteria 2fo2 nDm
having approximately the same OD600 values were used. Df  p 4
A ql
Electrochemical measurements were carried out in a conven-
tional three-electrode cell. The OCP, polarization curves and EIS where f0 is the resonant frequency of the fundamental mode of the
were measured using CompactStat Potentiostat (IviumStat, The quartz resonator, n is the harmonic number of the oscillation (here
Netherlands). The EIS measurements were carried out at 5 mV ap- n = 1), A is the piezoelectrically active surface area, q is the quartz
plied voltage at sinusoidal wave in a frequency range of 20 kHz crystal density (q = 2.684 g/cm3) and l is the shear modulus of
50 mHz. The EIS measurements were conducted at the end of quartz (l = 2.947  1011 g/cm s2). The theoretical thickness (Tf,
OCP measurements. The charge transfer resistance (Rct) was ob- cm) of the deposits was calculated using m/ (density g/cm3).
tained from the diameter of the semicircle in the Nyquist plot, After the experiments (5 h) the coated discs were taken from
and the values of percentage inhibition efciency, gi was using the mediums, rinsed with deionized water and dried under a
the equations below: stream of nitrogen gas. Surface morphology was inspected and
analyzed with a SEM at an acceleration voltage of 20 kV (JEOL
Rctinh  Rctuninh
gi  100 1 JSM-7000F Field Emission SEM-EDAX).

where Rct(uninh) and Rct(inh) are the uninhibited and inhibited charge 3. Results and discussion
transfer resistance, respectively. The polarization curves were ob-
tained by scanning from the open circuit potential towards Fig. 1 represents the potential time curves of the alloy coat-
200 mV anodically and cathodically at a scan rate of 1 mV/s. The ings as a function of time in sterile medium, medium inoculated
corrosion current density (Icorr) was determined graphically by with A. salmonicida and medium inoculated with C. michiganensis.
extrapolating the linear Tafel segment of the cathodic and anodic In Fig. 1, sterile medium exhibited lower open circuit potential
curves to the Ecorr (vs Ag/AgCl). The inhibition efciency, gp was cal- (c: 0.6 V (vs. Ag/AgCl)) than that in medium inoculated with C.
culated from polarization measurements according to the equation michiganensis (a: 0.57 V (vs. Ag/AgCl)) and in the medium inocu-
given below: lated with A. salmonicida (b: 0.58 V (vs. Ag/AgCl)). As a result,

Icorruninh  Icorrinh
gp  100 2 -0.35
Icorruninh a: C. michiganensis
b: A. salmonicida
where Icorr(uninh) and Icorr(inh) are the uninhibited and inhibited cor- -0.40 c: Medium
rosion current densities.
E / V vs. Ag/AgCl

All the recorded icorr values are converted into the corrosion rate
(A) in mm y1 (Millimeter per year; the penetration rate of corro-
sion through a metal/alloy) using the equation below [24]:
Ammy1 3:28  icorr  3
where M is the arithmetic average of the atomic weights of Ni a
(58.6 g/mol) and Co (58.9 g/mol) , n the number of electrons trans- -0.60 b
ferred in the corrosion reaction (n = 2) and d is the arithmetic aver- c
age of Ni (8.90 g cm3) and Co (8.96 g cm3)s densities. Average
atomic weights and densities were used in expression (3). 0 3000 6000 9000 12000 15000 18000
The QCM measurements were carried out in a Model 400A
Time sec
time-resolved quartz crystal microbalance (CH Instrument, USA)
linked computer with electrochemistry software (CHI400A). The Fig. 1. Variation of OCP of NiCo electrodes immersed in sterile and inoculated
working electrode used in QCM was gold AT-cut quartz crystal medium.
N.O. San et al. / Corrosion Science 65 (2012) 113118 115

-0.3 10
0 C. michiganensis
-0.4 -10 A. salmonicida

E / V vs. Ag/AgCl


m g / cm
-0.6 -40
-0.7 -60
A. salmonicida -90
-0.9 Medium
C. michiganensis -100
0 2000 4000 6000 8000 10000 12000 14000 16000 18000
-9 -8 -7 -6 -5 -4
-2 Time sec
log (i) / A cm
Fig. 3. Time dependence of mass changes of NiCo electrodes in sterile and
Fig. 2. The polarization curves of NiCo alloy coatings in sterile and inoculated
inoculated medium, at the room temperature.

Ecorr value of the medium shifted more negative value than Ecorr -9.0
A. salmonicida
values of the medium inoculated with bacteria. -8.0 Medium
Fig. 2 shows the polarization curves in the absence and pres- C. michiganensis
ence of two bacteria after 5 h of exposure. The electrochemical -7.0
parameters; corrosion potential, Ecorr, corrosion current, icorr, corro- -6.0
sion current density, Icorr, anodic Tafel slope ba, cathodic Tafel slope
-Z'' / k cm

bc, inhibition efciency from polarization measurements, gp, corro- -5.0

sion rates, A, and mass losses from QCM experiments were calcu-
lated and given in Table 1. As shown in Table 1, the corrosion 0.21 Hz

potential shifted to less negative values upon inoculation of the -3.0

bacteria. On the other hand, the inoculation of bacteria resulted
with a signicant decrease in corrosion current. From the results, 14.6 mHz
0.209 Hz
the corrosion current density value detected after 5 h of exposure -1.0
from the alloy coatings exposed to the medium was 12.29 lA/ 0.2 Hz
cm2. This value was much higher than all current density values
of the medium inoculated with two bacteria. This behavior showed 0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0
corrosion inhibition ability of bacteria on NiCo alloy coatings. The Z'' / k cm

Icorr value of medium inoculated with C. michiganensis has the high-

est gp% value and therefore, this bacterium was more protective Fig. 4. Impedance diagrams of NiCo alloy coatings immersed in sterile and
than A. salmonicida which caused less corrosion. In addition, the inoculated medium.
corrosion rates of the alloy coating discs in sterile medium and
medium inoculated with A. salmonicida and C. michiganensis were
calculated from the Eq. (3) are shown in Table 1. At the end of proportional to the corrosion rate, and decreased slightly over this
5 h long incubation period, the estimated corrosion rates were period. Results showed that the mass loss of NiCo alloy coatings
approximately ve times higher for the sterile medium in the medium was 96.9 lg/cm2 but after inoculation of bacteria
(132.61 mm y1) compared to the medium inoculated with C. the mass loss began to slow down. As a result, the mass loss values
michiganensis (25.42 mm y1). were 2.14 lg/cm2 for C. michiganensis and 6.10 lg/cm2 for A. sal-
QCM is capable of detecting small mass changes in the region of monicida. When we compared the corrosion abilities, A. salmonicida
ng/cm2 under controlled electrochemical conditions in aqueous was found to be more effective than C. michiganensis.
solutions [2628]. Fig. 3 shows the results of an experiment where Nyquist plots of NiCo alloy coatings immersed in sterile med-
the change at the resonance frequency of a quartz crystal was ium and medium inoculated with two bacteria for 5 h are shown in
monitored continuously during the corrosion process for 5 h. The Fig. 4. In addition Fig. 5 shows the physical structure of the surface
negative slopes of the mass vs time curves, as shown in Fig. 3, were lm on the coated alloy specimens and their corresponding

Table 1
Electrochemical parameters of NiCo alloy coatings corrosion inhibition in medium and medium inoculated with bacteria.

Ecorr, V vs Ag/AgCl icorr, mA Icorr, lA cm2 gp% Anodic Tafel slope Cathodic Tafel slope- Corrosion Rate QCM mass loss
ba, V dec1 bc, V dec1 (mm y1) (lg/cm2)
Medium 0.61 2.50 12.29 0.29 0.23 132.61 96.9
A. salmonicida 0.60 1.06 5.20 57.6 0.06 0.19 56.15 6.10
C. michiganensis 0.59 0.48 2.36 80.8 0.08 0.11 25.42 2.14
116 N.O. San et al. / Corrosion Science 65 (2012) 113118

Fig. 5. Equivalent circuits used for tting the impedance spectra based on a double-layer model of surface lm.

Table 2
Electrochemical model impedance parameters of the NiCo alloy coatings in the medium and inoculated medium with two bacteria.

Rs, O Rp, k O cm2 Rct, k O cm2 gi% QCPE, X1 cm2  105 g Qdl, X1 cm2  105 g
Medium 441.2 7.6 19.4 24.0 0.85 24.9 0.7
A. salmonicida 830.2 19.1 8.4 56.6 18.1 0.8 44.6 0.09
C. michiganensis 1159.9 13.5 3.2 83.5 5.6 0.9 13.5 0.91

Fig. 6. SEM micrographs and EDS spectrum of NiCo alloy coated surfaces exposed to sterile medium and medium containing two bacteria for 5 h: (a,b) NiCo alloy coating in
sterile medium (c,d) NiCo alloy coating in medium containing A. salmonicida (e,f) NiCo alloy coating in medium containing C. michiganensis.
N.O. San et al. / Corrosion Science 65 (2012) 113118 117

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