Microbiologically Influenced Corrosion of 2707 Hyper

Scientific Reports volume 6, Article number: 20190 (2016) Cite this article 6601 Accesses 108 Citations 6 Altmetric Metrics details Microbiologically Influenced Corrosion (MIC) is a serious problem in

Scientific Reports volume 6, Article number: 20190 (2016) Cite this article

6601 Accesses

108 Citations

6 Altmetric

Metrics details

Microbiologically Influenced Corrosion (MIC) is a serious problem in many industries because it causes huge economic losses. Due to its excellent resistance to chemical corrosion, 2707 hyper duplex stainless steel (2707 HDSS) has been used in the marine environment. However, its resistance to MIC was not experimentally proven. In this study, the MIC behavior of 2707 HDSS caused by the marine aerobe Pseudomonas aeruginosa was investigated. Electrochemical analyses demonstrated a positive shift in the corrosion potential and an increase in the corrosion current density in the presence of the P. aeruginosa biofilm in the 2216E medium. X-ray photoelectron spectroscopy (XPS) analysis results showed a decrease in Cr content on the coupon surface beneath the biofilm. The pit imaging analysis showed that the P. aeruginosa biofilm caused a largest pit depth of 0.69 μm in 14 days of incubation. Although this was quite small, it indicated that 2707 HDSS was not completely immune to MIC by the P. aeruginosa biofilm.

Duplex stainless steel (DSS) is widely used in various industries due to its desirable combination of excellent mechanical properties and corrosion resistance1,2. However, localized pitting corrosion can still occur and it affects the integrity of this kind of steel3,4. DSS is not immune to microbiologically influenced corrosion (MIC)5,6. Despite a very broad range of applications for DSS, there are still environments where the corrosion resistance of DSS is inadequate for long-term services. This means, more expensive materials with higher corrosion resistance are needed. Jeon et al.7 found that even super duplex stainless steels (SDSSs) exhibit some limits in corrosion resistance. Therefore, hyper duplex stainless steels (HDSSs) with a higher corrosion resistance are needed in some applications. This led to the development of highly alloyed HDSSs.

The corrosion resistance of DSS is determined by the ratio of the α-phase and γ-phase and by the Cr, Mo and W depleted regions that are adjacent to the secondary phases8,9,10. HDSS contains high levels of Cr, Mo and N11, resulting in its excellent corrosion resistance and a high value (45–50) of pitting resistance equivalent number (PREN), which is calculated from wt.% Cr + 3.3 (wt.% Mo + 0.5 wt.% W) + 16 wt.% N12. Its excellent corrosion resistance properties rely on a well-balanced composition with approximately 50% ferrite (α) and 50% austenite (γ) phases, that offer HDSS improved mechanical properties and higher chloride corrosion resistance compared with conventional DSS13. The improved corrosion resistance extends the use of HDSS in more aggressive chloride environments, such as marine environments.

MIC is a major problem in many industries such as oil and gas, as well as water utilities14. MIC accounts for 20% of all corrosion damages15. MIC is bioelectrochemical corrosion that can be observed in many environments16. Biofilms formed on a metal surface will change the electrochemical conditions and thus influencing the corrosion processes. It is widely accepted that MIC corrosion is caused by biofilms14. Electrogenic microbes corrode metals in order to obtain maintenance energy for survival17. The most recent MIC research suggested that EET (extracellular electron transfer) is a rate-limiting factor in MIC caused by electrogenic microbes. Zhang et al.18 demonstrated that an electron mediator accelerated the electron transfer between sessile Desulfovibrio vulgaris cells and 304 stainless steel, resulting in a much more severe MIC attack. Enning et al.19 and Venzlaff et al.20 showed that a corrosive sulfate-reducing bacterium (SRB) biofilm was capable of directly uptaking electrons from the metal matrix, resulting in serious pitting corrosion.

DSS is known to be susceptible to MIC in environments containing SRB, iron reducing bacteria (IRB), etc21. These bacteria cause localized pitting corrosion on DSS surfaces under biofilms22,23. Unlike DSS, very little is known for the MIC of HDSS24.

Pseudomonas aeruginosa is a Gram-negative motile rod bacterium widely distributed in nature25. P. aeruginosa is also a predominant group of micro-organisms in marine environments causing MIC to steels26. The genus Pseudomonas is closely involved in corrosion processes and has been recognized as the pioneer colonizer in the process of biofilm formation27. Mahat et al.28 and Yuan et al.29 demonstrated that P. aeruginosa has the propensity to increase the corrosion rates of mild steels and alloys in aquatic environments.

The main aim of this work was to investigate the MIC characteristics of 2707 HDSS caused by the marine aerobe P. aeruginosa using electrochemical methods, surface analysis techniques and corrosion product analyses. Electrochemical studies, including open circuit potential (OCP), linear polarization resistance (LPR), electrochemical impedance spectroscopy (EIS) and potential dynamic polarization were performed to study the MIC behavior of 2707 HDSS. The energy dispersive spectrometer (EDS) analysis was undertaken to find the chemical elements on the corroded surface. In addition, the X-ray photoelectron spectroscopy (XPS) analysis was used to determine the stability of the oxide film passivity under the influence of a marine environment containing P. aeruginosa. The pit depth was measured under a confocal laser scanning microscope (CLSM).

Table 1 lists the chemical composition of 2707 HDSS. Table 2 shows that 2707 HDSS has excellent mechanical properties with yield strength of 650 MPa. Figure 1 reveals the optical microstructures of the solution heat-treated 2707 HDSS. Elongated bands of austenite and ferrite phases without secondary phases can be seen in the microstructure that contains approximately 50% austenite and 50% ferrite phases.

Microstructure of 2707 HDSS:

(A) ferrite and (B) austenite.

Figure 2a shows open circuit potential (Eocp) vs. exposure time data for 2707 HDSS in the abiotic 2216E medium and in the P. aeruginosa broth at 37 °C for 14 days. It reveals that the most and significant variation in Eocp occurred in the initial 24 h. The Eocp values in both cases peaked around −145 mV (vs. SCE) at about 16 h and then dropped sharply, reaching −477 mV (vs. SCE) and −236 mV (vs. SCE) for the abiotic coupon and the P. aeruginosa coupon, respectively. After 24 h, the Eocp value of 2707 HDSS remained relatively stable at −228 mV (vs. SCE) for the P. aeruginosa coupon, while the corresponding value for the abiotic coupon was approximately −442 mV (vs. SCE). The Eocp in the presence of P. aeruginosa was considerably lower.

Electrochemical tests for 2707 HDSS coupons in the abiotic medium and in the P. aeruginosa broth at 37 °C:

(a) variation of Eocp with exposure time, (b) polarization curves on the 14th day, (c) variation of Rp with exposure time and (d) variation of icorr with exposure time.

The values of the electrochemical corrosion parameters of 2707 HDSS coupons after exposure for 14 days in the abiotic medium as well as in the P. aeruginosa inoculated medium are listed in Table 3. The tangential lines of the anodic and cathodic curves were extrapolated to reach an intersection point to yield corrosion current density (icorr), corrosion potential (Ecorr) and Tafel slopes (βα and βc) following the standard approach30,31.

As shown in Figure 2b, compared with the abiotic curves, the P. aeruginosa curves shifted upward leading to an increase of Ecorr. The icorr value, that was directly proportional to the corrosion rate, increased to 0.328 μA cm−2 for the P. aeruginosa coupon, four times greater than that for the abiotic coupon (0.087 μA cm−2).

LPR is a classical electrochemical method for fast corrosion analysis that is non-destructive. It is also used to study MIC32. Figure 2c shows the variation of the polarization resistance (Rp) as a function of exposure time. A higher Rp value means less corrosion. During the initial 24 h, the Rp of 2707 HDSS reached a maximum value of 1955 kΩ cm2 for the abiotic coupon and 1429 kΩ cm2 for the P. aeruginosa coupon. Figure 2c also shows that the Rp value fell quickly after one day and then remained relatively unchanged for the next 13 days. The Rp value for the P. aeruginosa coupon was around 40 kΩ cm2, much lower than the 450 kΩ cm2 value for the abiotic coupon.

The icorr value is directly proportional to the uniform corrosion rate. Its value can be calculated from the Stern-Geary equation below,

Following Zou et al.33, the Tafel slope B in this work was assumed a typical value of 26 mV/dec. Figure 2d shows that icorr remained relatively stable for the abiotic 2707 coupon while it fluctuated considerably for the P. aeruginosa coupon that had a large jump after the initial 24 h. The icorr value for the P. aeruginosa coupon was one order of magnitude higher than that of the abiotic control. This trend agreed with the polarization resistance result.

EIS is another nondestructive technique to characterize electrochemical reactions at a corrosion interface34. Impedance spectra and calculated capacitance values of coupons exposed to the abiotic medium and the P. aeruginosa solution, Rb the resistance of passive film/biofilm formed on the coupon surface, Rct the charge transfer resistance, Cdl the capacitance of the electrical double layer (EDL) and QCPE the constant phase element (CPE) parameter. These parameters were further analyzed by fitting data with an equivalent electrical circuit (EEC) model.

Figure 3 shows the typical Nyquist diagrams (a and b) and Bode plots (a′ and b′) at different incubation times for 2707 HDSS coupons in the abiotic medium and in the P. aeruginosa broth. The diameters of the Nyquist loops decreased in the presence of P. aeruginosa. The Bode plot (Figure 3b′) shows an increase of the total impedance magnitudes. The information of the relaxation time constants can be provided by the phase maxima. Figure 4 shows the physical structure and their corresponding EECs based on a single-layer (a) and a double-layer (b). CPE was introduced into the EEC models. Its admittance and impedance, respectively, are expressed below:

The Nyquist and Bode plots of 2707 HDSS coupons with and without exposure to P. aeruginosa:

(a,a′) 2707 HDSS in the abiotic medium and (b,b′) 2707 HDSS in the P. aeruginosa broth.

Two physical models and the corresponding equivalent circuits used for fitting the impedance spectra of 2707 HDSS coupons:

(a) in the abiotic medium and (b) in the P. aeruginosa broth.

where Y0 is the magnitude of the CPE, j the imaginary number or (−1)1/2, ω the angular frequency and n the CPE power index which is less than unity35. The inverse of the charge-transfer resistance (i.e., 1/Rct) corresponds to the corrosion rate. A smaller Rct means a faster corrosion rate27. After 14 days of incubation, the Rct of the P. aeruginosa coupon reached 32 kΩ cm2, much smaller than the 489 kΩ cm2 for the abiotic coupon (Table 4).

The CLSM images and SEM images in Figure 5 clearly indicate that the biofilm coverage on the 2707 HDSS coupon surface after 7 days was dense. However, after 14 days the biofilm coverage was sparse and some dead cells appeared. Table 5 shows the biofilm thickness on the 2707 HDSS coupon after exposure to P. aeruginosa for 7 and 14 days. The largest biofilm thickness changed from 23.4 μm after 7 days to 18.9 μm after 14 days. The average biofilm thickness also confirmed the trend. It decreased from 22.2 ± 0.7 μm after 7 days to 17.8 ± 1.0 μm after 14 days.

The morphology of P. aeruginosa biofilms on coupon surfaces:

(a) the 3-D CLSM image after 7 days, (b) the 3-D CLSM image after 14 days, (c) the SEM image after 7 days and (d) the SEM image after 14 days.

Chemical elements in the biofilm and corrosion products on the coupon exposed to P. aeruginosa for 14 days were revealed by EDS. Figure 6 shows the amounts of C, N, O, P in the biofilm and corrosion products were much higher than those in the bare metal because these elements were associated with the biofilm and its metabolic products. Cr and Fe are needed by microorganisms only at trace levels. The high levels of Cr and Fe in the biofilm and corrosion products on the coupon surface suggest loss of the elements by the metal matrix due to corrosion.

Results of EDS analysis of the 2707 HDSS surface after exposure to P. aeruginosa for 14 days.

Pitting corrosion was observed with and without P. aeruginosa in the 2216E medium after 14 days. Before incubation, the coupon surfaces were smooth and had no defects (Figure 7a). After incubation and removing the biofilm and corrosion products, the deepest pits on the coupon surfaces were examined under CLSM as shown in Figure 7b and c. No significant pits were found on the abiotic control coupon surface (largest pit depth 0.02 μm). The largest pit depth caused by P. aeruginosa was 0.52 μm after 7 days and 0.69 μm after 14 days and the average largest pit depth based on 3 coupons (with 10 largest pit depth values per coupon selected) reached 0.42 ± 0.12 μm and 0.52 ± 0.15 μm, respectively (Table 5). These pit depth values were small, but significant.

CLSM analysis of pits on coupon surfaces:

(a) before exposure, (b) in the abiotic medium for 14 days and (c) in the P. aeruginosa broth for 14 days.

Figure 8 shows the XPS spectra of different coupon surfaces and the chemical composition of each surface analysis is summarized in Table 6. In Table 6, the atomic percentages of Fe and Cr in the presence of P. aeruginosa (samples A and B) were much lower than those for the abiotic control coupons (samples C and D). For the P. aeruginosa coupons, the Cr 2p core-level spectrum was curve-fitted into four peak components at binding energy (BE) values of 574.4, 576.6, 578.3 and 586.8 eV, which were attributable to Cr, Cr2O3, CrO3 and Cr(OH)3, respectively (Figure 9a and b). For the abiotic coupons, the Cr 2p core-level spectrum contained two dominant peaks for Cr (BE at 573.80 eV) and Cr2O3 (BE at 575.90 eV) in Figure 9c and d, respectively. The most noticeable difference between the abiotic coupons and the P. aeruginosa coupons was the presence of Cr6+ beneath the biofilm and a higher relative portion of Cr(OH)3 (BE at 586.8 eV).

The wide XPS spectra for the surfaces of 2707 HDSS coupons in the both media for 7 days and 14 days, respectively.

The high resolution XPS spectra of Cr 2p for 2707 HDSS:

(a) with exposure to P. aeruginosa for 7 days, (b) with exposure to P. aeruginosa for 14 days, (c) in the abiotic medium for 7 days and (d) in the abiotic medium for 14 days.

HDSSs exhibit a high level of corrosion resistance in most environments. Kim et al.2 reported that UNS S32707 HDSS is defined as a highly alloyed DSS with a PREN in excess of 45. The 2707 HDSS coupons in this work had a PREN value of 49. This is due to its high Cr content and elevated Mo and Ni levels, which are beneficial in acid and high chloride environments. Furthermore, the well-balanced composition and defect-free microstructure are useful for structural stability and corrosion resistance. Despite its superior chemical corrosion resistance, however, the experimental data in this work demonstrated that 2707 HDSS was not completely immune to the MIC by the P. aeruginosa biofilm.

The electrochemical results revealed that the corrosion rate of 2707 HDSS significantly increased in the P. aeruginosa broth after 14 days compared to the abiotic medium. In Figure 2a, a decrease of Eocp was observed in both the abiotic medium and the P. aeruginosa broth in the initial 24 h. Afterwards, the biofilm completed the coverage of the coupon surface, Eocp became relatively stable36. However, the biotic Eocp was at a much higher level than the abiotic Eocp. It is reasonable to believe that the difference was due to the formation of the P. aeruginosa biofilm. In Figure 2d, the icorr value of 2707 HDSS reached 0.627 μA cm−2 in the presence of P. aeruginosa, one order of magnitude higher than that for the abiotic control (0.063 μA cm−2), which was consistent with the Rct values measured by EIS. In the first few days, the impedance value increased in the P. aeruginosa broth due to the attachment of P. aeruginosa cells and the formation of the biofilm. However, the impedance then decreased when the biofilm fully covered the coupon surface. The protective layer was first attacked because of the formation of the biofilm and metabolites of the biofilm. Consequently, the corrosion resistance decreased with time and the P. aeruginosa attachment caused the localized corrosion. The tendency in the abiotic medium was different. The corrosion impedance of the abiotic control was much higher than the corresponding value of the coupon exposed to the P. aeruginosa broth. Furthermore, for the abiotic samples, the Rct value of 2707 HDSS reached 489 kΩ cm2 on the 14th day, fifteen times larger than that in the presence of P. aeruginosa (32 kΩ cm2). Thus, 2707 HDSS possessed excellent corrosion resistance in the sterile environment, but it was not immune to the MIC attack by the P. aeruginosa biofilm.

These results were also observed from the polarization curves in Figure 2b. The anodic branch was attributed to the formation of P. aeruginosa biofilm and the metal oxidation reaction. Meanwhile the cathodic reaction was the reduction of oxygen. The presence of P. aeruginosa greatly increased the corrosion current density, which was approximately one order of magnitude higher than the abiotic control. It suggested that the P. aeruginosa biofilm increased the localized corrosion of 2707 HDSS. Yuan et al.29 found that the 70/30 Cu-Ni alloy corrosion current density increased under the attack of a P. aeruginosa biofilm. It could be due to the P. aeruginosa biofilm’s biocatalysis of the reduction of oxygen37. This observation could explain the MIC of 2707 HDSS in this work as well. It was also possible that the aerobic biofilm made oxygen less available underneath it. Thus, the failure to repassivate the metal surface by oxygen could be a contributing factor in the MIC in this work.

Dickinson et al.38 proposed that the rates of chemical and electrochemical reactions can be directly affected by the metabolic activities of the sessile bacteria on a coupon surface and the nature of corrosion products. As seen in Figure 5 and Table 5, both the cell number and biofilm thickness decreased after 14 days. This can be reasonably explained that after 14 days, most of the sessile cells on the 2707 HDSS surface were dead due to the exhaustion of the nutrients in the 2216E medium or the toxic metal ions released from the 2707 HDSS matrix. This was a limitation of batch experiments.

In this work, the P. aeruginosa biofilm promoted the localized depletion of Cr and Fe underneath the biofilm on the 2707 HDSS surface (Figure 6). In Table 6, compared with sample C, there was a decrease of Fe and Cr in sample D, indicating that dissolution Fe and Cr caused by the P. aeruginosa biofilm continued beyond the first 7 days. The 2216E medium was used to simulate the marine environment. It contained 17700 ppm Cl−, comparable to that in the natural seawater. The presence of 17700 ppm Cl− was the main reason responsible for the decrease of Cr from the 7-day and 14-day abiotic samples analyzed by XPS. Compared with the P. aeruginosa coupons, the dissolution of Cr in abiotic coupons was much smaller due to the strong Cl− resistance of 2707 HDSS in an abiotic environment. Figure 9 shows that the presence of Cr6+ in the passivation film. It might be involved in the removal of Cr from the steel surface by the P. aeruginosa biofilm as suggested by Chen and Clayton39.

The pH values of the culture medium before and after incubation were 7.4 and 8.2, respectively, due to bacterial growth. Thus, underneath the P. aeruginosa biofilm, organic acid corrosion was unlikely a contributing factor in this work due to the relatively high pH values in the bulk medium. The pH value of the abiotic control medium didn’t change significantly (from the initial 7.4 to the final 7.5) in the 14-day test duration. The increase of pH in the inoculated medium after incubation was due to the metabolic activities of P. aeruginosa, which was found to have the same impact on pH in the absence of coupons.

As shown in Figure 7, the maximum pit depth caused by the P. aeruginosa biofilms was 0.69 μm, which was much larger than that in the abiotic medium (0.02 μm). This was consistent with the electrochemical data above. The 0.69 μm pit depth was more than ten times smaller than the 9.5 μm value reported for 2205 DSS under the same conditions40. These data proved that 2707 HDSS exhibited a better MIC resistance compared with 2205 DSS. This should not be a surprise because 2707 HDSS has a higher Cr level that offers more durable passivity, making it more difficult for P. aeruginosa to depassivate and initiate pitting corrosion due to the balanced phase structure without harmful secondary precipitates41.

In conclusion, MIC pitting corrosion was found on the surface of 2707 HDSS in the P. aeruginosa broth compared to negligible pitting in the abiotic medium. This work demonstrated that 2707 HDSS had much better MIC resistance than 2205 DSS, but was not completely immune to MIC due to the P. aeruginosa biofilm. These findings are useful in the selection of a suitable stainless steel for the marine environment and in the estimation of service life span.

The coupons of 2707 HDSS were provided by School of Metallurgy, Northeastern University (NEU) in Shenyang, China. The elemental composition of 2707 HDSS was shown in Table 1, which was analyzed by the Department of Materials Analysis and Testing of NEU. All specimens were solution-treated at 1180 °C for 1 h. Prior to the corrosion tests, coin-shaped 2707 HDSS with a top exposed surface area of 1 cm2 were polished to 2000 grit with silicon carbide papers and then further polished with a 0.05 μm Al2O3 powder suspension. The side and bottom surface were protected by an inert paint. After drying, the coupons were rinsed with sterile deionized water and sterilized with 75% (v/v) ethanol for 0.5 h. They were then air dried under ultraviolet (UV) light for 0.5 h before use.

The marine P. aeruginosa MCCC 1A00099 strain was purchased from the Marine Culture Collection of China (MCCC), Xiamen, China. The marine 2216E liquid medium (Qingdao Hope Bio-technology Co., Qingdao, China) was used to culture the P. aeruginosa at 37 °C aerobically in 250 ml flasks and in a 500 ml electrochemical glass cell. The culture medium contained (g/L): 19.45 NaCl, 5.98 MgCl2, 3.24 Na2SO4, 1.8 CaCl2, 0.55 KCl, 0.16 Na2CO3, 0.08 KBr, 0.034 SrCl2, 0.08 SrBr2, 0.022 H3BO3, 0.004 NaSiO3, 0.0024 NaF, 0.0016 NH4NO3, 0.008 NaH2PO4, 5.0 peptone, 1.0 yeast extract and 0.1 ferric citrate. It was autoclave-sterilized at 121 °C for 20 min prior to inoculation. A hemocytometer under a light microscope at 400× magnification was used to enumerate sessile and planktonic cells. The initial planktonic P. aeruginosa cell concentration immediately after inoculation was approximately 106 cells/ml.

Electrochemical tests were carried out in a classical three-electrode glass cell with a culture medium volume of 500 ml. A platinum sheet and a saturated calomel electrode (SCE) connected to the reactor via a Luggin capillary filled with a salt bridge as the counter and reference electrodes, respectively. To create a working electrode, a rubber-coated copper wire was connected to each coupon and covered with epoxy resin leaving an exposed one-sided surface area of approximately 1 cm2 for this working electrode. During electrochemical measurements, samples were placed in the 2216E medium and maintained at a constant incubation temperature (37 °C) in a water bath. The OCP, LPR, EIS and potential dynamic polarization data were measured with an Autolab potentiostat (Reference 600TM, Gamry Instruments, Inc., USA). LPR tests were recorded at a scan rate of 0.125 mV s−1 in the range of −5 and 5 mV versus Eocp and the sampling frequency was 1 Hz. EIS was performed under a steady-state Eocp using a 5 mV applied voltage at sinusoidal wave in a frequency range of 0.01 to 10,000 Hz. Before the potential scan, the electrodes were in the open circuit mode until a steady free corrosion potential value reached42. The polarization curves were then run at a scan rate of 0.166 mV/s from −0.2 to 1.5 V vs. Eocp. Each test was repeated three times with and without P. aeruginosa.

The coupons for the metallographic analysis were mechanically polished with wet SiC paper of 2000 grit and then further polished with a 0.05 μm Al2O3 powder suspension to enable optical observations to be made. Metallographic analysis was carried out using an optical microscopy. The coupons were etched with a 10 wt.% potassium hydroxide solution43.

After incubation, a coupon was washed with a phosphate buffer saline (PBS) solution (pH 7.4 ± 0.2) three times and then fixed with 2.5% (v/v) glutaraldehyde for 10 h to fix the biofilm. It was subsequently dehydrated with a graded series (50%, 60%, 70%, 80%, 90%, 95% and 100% v/v) of ethanol before air-drying. Finally, the coupon surface was sputter-coated with a gold film to provide conductivity for SEM observation44. SEM images focused on spots with the most sessile P. aeruginosa cells on each coupon surface. The EDS analysis was undertaken to find the chemical elements. A Zeiss confocal laser scanning microscope (CLSM) (LSM 710, Zeiss, Germany) was used to measure the pit depth. To observe the corrosion pits underneath a biofilm, the coupon was first cleaned according to the Chinese National Standards (CNS) GB/T4334.4-2000 to remove the corrosion products and biofilms on the coupon surface.

An X-ray photoelectron spectroscopy (XPS, ESCALAB250 surface analysis system, Thermo VG, USA) analysis was performed using a monochromatic X-ray source (an aluminum Kα line of 1500 eV energy and 150 W power) within the wide binding energy range of 0–1350 eV under standard conditions. The high resolution spectra were recorded using 50 eV pass energy and 0.2 eV step.

The coupons after incubation were taken out and gently rinsed with PBS (pH 7.4 ± 0.2) for 15 s45. In order to observe the bacterial viability of the biofilms on the coupon, LIVE/DEAD BacLight Bacterial Viability Kits (Invitrogen, Eugene, OR, USA) were used to stain the biofilms. The kits have two fluorescent dyes, the green-fluorescent SYTO-9 dye and the red-fluorescent propidium iodide (PI) dye. Under CLSM, the dots with fluorescent green and red represented the live and dead cells, respectively. When staining, 1 ml mixture containing 3 μl SYTO-9 and 3 μl PI solutions were incubated for 20 min in the dark at room temperature (23 oC). After that, a Nikon CLSM machine (C2 Plus, Nikon, Japan) was used to observe the stained sample at at two wave lengths (488 nm for living cells and 559 nm for dead cells). The biofilm thickness was measured in the 3-D scanning mode.

How to cite this article: Li, H. et al. Microbiologically Influenced Corrosion of 2707 Hyper-Duplex Stainless Steel by Marine Pseudomonas aeruginosa Biofilm. Sci. Rep. 6, 20190; doi: 10.1038/srep20190 (2016).

Zanotto, F., Grassi, V., Balbo, A., Monticelli, C. & Zucchi, F. Stress corrosion cracking of LDX 2101 duplex stainless steel in chloride solutions in the presence of thiosulphate. Corros. Sci. 80, 205–212 (2014).

CAS Google Scholar

Kim, S. T., Jang, S. H., Lee, I. S. & Park, Y. S. Effects of solution heat-treatment and nitrogen in shielding gas on the resistance to pitting corrosion of hyper duplex stainless steel welds. Corros. Sci. 53, 1939–1947 (2011).

CAS Google Scholar

Shi, X., Avci, R., Geiser, M. & Lewandowski, Z. Comparative study in chemistry of microbially and electrochemically induced pitting of 316L stainless steel. Corros. Sci. 45, 2577–2595 (2003).

CAS Google Scholar

Luo, H., Dong, C. F., Li, X. G. & Xiao, K. The electrochemical behavior of 2205 duplex stainless steel in alkaline solutions with different pH in the presence of chloride. Electrochim. Acta. 64, 211–220 (2012).

CAS Google Scholar

Little, B. J., Lee, J. S. & Ray, R. I. The influence of marine biofilms on corrosion: A concise review. Electrochim. Acta. 54, 2–7 (2008).

CAS Google Scholar

Liang, C., Wang, H. & Huang, N. Effects of Sulphate-reducing bacteria on corrosion behaviour of 2205 duplex stainless steel. J. Iron. Steel. Res. 21, 444–450 (2014).

CAS Google Scholar

Jeon, S. H., Kim, H. J. & Park, Y. S. Effects of inclusions on the precipitation of chi phases and intergranular corrosion resistance of hyper duplex stainless steel. Corros. Sci. 87, 1–5 (2014).

CAS Google Scholar

Deng, B. et al. Effect of thermal cycles on the corrosion and mechanical properties of UNS S31803 duplex stainless steel. Corros. Sci. 51, 2969–2975 (2009).

CAS Google Scholar

Kim, J. S. et al. Mechanism of localized corrosion and phase transformation of tube-to-tube sheet welds of hyper duplex stainless steel in acidified chloride environments. Mater. Trans. 12, 2166–2174 (2012).

Google Scholar

Deng, B. et al. Evaluation of localized corrosion in duplex stainless steel aged at 850 °C with critical pitting temperature measurement. Electrochim. Acta. 54, 2790–2794 (2009).

CAS Google Scholar

Pilhagen, J., Sieurin, H. & Sandstrom, R. Fracture toughness of a welded super duplex stainless steel. Mat. Sci. Eng. A-Struct. 606, 40–45 (2014).

CAS Google Scholar

Lee, J. S., Fushimi, K., Nakanishi, T., Hasegawa, Y. & Park, Y. S. Corrosion behavior of ferrite and austenite phases on super duplex stainless steel in a modified green-death solution. Corros. Sci. 89, 111–117 (2014).

CAS Google Scholar

Kim, H. J., Jeon, S. H., Kim, S. T. & Park, Y. S. Influence of the shielding gas composition on the passive film and erosion corrosion of tube-to-tube sheet welds of hyper duplex stainless steel. Corros. Sci. 91, 140–150 (2015).

CAS Google Scholar

Xu, D., Li, Y., Song, F. & Gu, T. Laboratory investigation of microbiologically influenced corrosion of C1018 carbon steel by nitrate reducing bacterium Bacillus licheniformis. Corros. Sci. 77, 385–390 (2013).

CAS Google Scholar

Alabbas, F. M. et al. Influence of sulfate reducing bacterial biofilm on corrosion behavior of low-alloy, high-strength steel (API-5L X80). Int. Biodeterior. Biodegrad. 78, 34–42 (2013).

CAS Google Scholar

Hamzah, E., Hussain, M. Z., Ibrahim, Z. & Abdolahi, A. Influence of Pseudomonas aeruginosa bacteria on corrosion resistance of 304 stainless steel. Corros. Eng. Sci. Techn. 48, 116–120 (2013).

CAS Google Scholar

Xu, D. & Gu, T. Carbon source starvation triggered more aggressive corrosion against carbon steel by Desulfovibrio vulgaris biofilm. Int. Biodeter. Biodegr. 91, 74–81 (2014).

CAS Google Scholar

Zhang, P., Xu, D., Li, Y., Yang, K. & Gu, T. Electron mediators accelerate the microbiologically influenced corrosion of 304 stainless steel by Desulfovibrio vulgaris biofilm. Bioelectrochemistry. 101, 14–21 (2015).

CAS PubMed Google Scholar

Venzlaff, H. et al. Accelerated cathodic reaction in microbial corrosion of iron due to direct electron uptake by sulfate-reducing bacteria. Corros. Sci. 66, 88–96 (2013).

CAS Google Scholar

Enning, D. et al. Marine sulfate-reducing bacteria cause serious corrosion of iron under electroconductive biogenic mineral crust: Microbial iron corrosion under electroconductive crust. Environ. Microbiol. 14, 1772–1787 (2012).

CAS PubMed PubMed Central Google Scholar

Xu, C., Zhang, Y., Cheng, G. & Zhu, W. Pitting corrosion behavior of 316L stainless steel in the media of sulphate-reducing and iron-oxidizing bacteria. Mater. Charact. 59, 245–255 (2008).

CAS Google Scholar

Shi, X. et al. Comparative study in chemistry of microbially and electrochemically induced pitting of 316L stainless steel. Corros. Sci. 45, 2577–2595 (2003).

CAS Google Scholar

Mansfeld, F. The interaction of bacteria and metal surfaces. Electrochim. Acta. 52, 7670–7680 (2007).

CAS Google Scholar

Antony, P. J., Singh Raman, R. K., Mohanram, R. & Kumar, P. Influence of thermal aging on sulfate-reducing bacteria (SRB)-influenced corrosion behaviour of 2205 duplex stainless steel. Corros. Sci. 50, 1858–1864 (2008).

CAS Google Scholar

Mansouri, H., Alavi, S. A. & Yari, M. A. Study of Pseudomonas aeruginosa bacteria in microbial corrosion. 2nd International Conference on Chemical, Ecology and Environmental Sciences. Singapore April, 28–29 (2012).

Stott, J. F. D. What progress in the understanding of microbially induced corrosion has been made in the last 25 years? A personal view point. Corros. Sci. 35, 667–673 (1993).

CAS Google Scholar

San, N. O., Nazir, H. & Donmez, G. Microbially influenced corrosion and inhibition of nickel-zinc and nickel-copper coatings by Pseudomonas aeruginosa. Corros. Sci. 79, 177–183 (2014).

CAS Google Scholar

Mahat, M. M. et al. A preliminary study on Microbiologically influenced corrosion (MIC) of mild steel by Pseudomonas aeruginosa by using infinite focus microscope (IFM). AIP. Conf. Proc. 1455, 117 (2012).

ADS CAS Google Scholar

Yuan, S. J., Choong, A. M. F. & Pehkonen, S. O. The influence of the marine aerobic Pseudomonas strain on the corrosion of 70/30 Cu-Ni alloy. Corros. Sci. 49, 4352–4385 (2007).

CAS Google Scholar

Yuan, S. Y., Liang, B., Zhao, Y. & Pehkonen, S. O. Surface chemistry and corrosion behavior of 304 stainless steel in simulated seawater containing inorganic sulphide and sulphate-reducing bacteria. Corros. Sci. 74, 353–366 (2013).

CAS Google Scholar

Yuan, S. J. et al. Biocorrosion behavior of titanium oxide/butoxide-coated stainless steel. J. Electrochem. Soc. 155, 196–210 (2008).

Google Scholar

Millard, S. G., Law, D., Bungey, J. H. & Cairns, J. Environmental influences on linear polarisation corrosion rate measurement in reinforced concrete. NDT. & E. Int. 34, 409–417 (2001).

CAS Google Scholar

Zou, Y., Wang, J. & Zheng, Y. Y. Electrochemical techniques for determining corrosion rate of rusted steel in seawater. Corros. Sci. 53, 208–216 (2011).

CAS Google Scholar

Mansfeld, F., Liu, G., Xiao, H., Tsai, C. H. & Little, B. J. The corrosion behavior of copper alloys, stainless steels and titanium in seawater. Corros. Sci. 36, 2063–2095 (1994).

CAS Google Scholar

Wu, X., Ma, H., Chen, S., Xu, Z. & Sui, A. General equivalent circuits for faradaic electrode processes under electrochemical reaction control. J. Electrochem. Soc. 146, 1847–1853 (1999).

CAS Google Scholar

Moradi, M., Song, Z., Yang, L., Jiang, J. & He, J. Effect of marine Pseudoalteromonas sp. on the microstructure and corrosion behaviour of 2205 duplex stainless steel. Corros. Sci. 84, 103–112 (2014).

CAS Google Scholar

Cournet, A., Berge, M., Roques, C., Bergel, A. & Delia, M. L. Electrochemical reduction of oxygen catalyzed by Pseudomonas aeruginosa. Electrochim. Acta. 55, 4902–4908 (2010).

CAS Google Scholar

Dickinson, W. H. & Lewandowski, Z. Electrochemical concepts and techniques in the study of stainless steel ennoblement. Biodegradation. 9, 11–21 (1998).

CAS PubMed Google Scholar

Chen, G. & Clayton, C. R. The influence of sulfate-reducing bacteria on the passivity of type 317L austenitic stainless steel. J. Electrochem. Soc. 145, 1914–1922 (1998).

CAS Google Scholar

Xia, J. et al. Laboratory investigation of the microbiologically influenced corrosion (MIC) resistance of a novel Cu-bearing 2205 duplex stainless steel in the presence of an aerobic marine Pseudomonas aeruginosa biofilm. Biofouling. 31, 481–492 (2015).

CAS PubMed Google Scholar

Antony, P. J., Singh Raman, R. K., Raman, R. & Kumar, P. Role of microstructure on corrosion of duplex stainless steel in presence of bacteria activity. Corros. Sci. 52, 1404–1412 (2010).

CAS Google Scholar

Atta, N. F., Fekry, A. M. & Hassaneen, H. M. Corrosion inhibition, hydrogen evolution and antibacterial properties of newly synthesized organic inhibitors on 316L stainless steel alloy in acid medium. Int. J. Hydrogen. Energ. 36, 6462–6471 (2011).

CAS Google Scholar

Ren, L., Nan, L. & Yang, K. Study of copper precipitation behavior in a Cu-bearing austenitic antibacterial stainless steel. Mater. Des. 32, 2374–2379 (2011).

CAS Google Scholar

Wen, J., Zhao, K., Gu, T. & Raad, I. I. A green biocide enhancer for the treatment of sulfate-reducing bacteria (SRB) biofilms on carbon steel surfaces using glitaraldehyde. Int. Biodeter. Biodegr. 63, 1102–1106 (2009).

CAS Google Scholar

Jin, Y. et al. The use of new probes and stains for improved assessment of cell viability and extracellular polymeric substances in Candida albicans biofilms. Mycopathologia. 159, 353–360 (2005).

CAS PubMed Google Scholar

Download references

This work was finically supported by the High Technology Research and Development Program of China (No. 2015AA034301), the National Natural Science Foundation (No. 51501203), the National Key Technology Research and Development Program of the Ministry of Science and Technology of China (No. 2012BAE04B01), the Program for New Century University Excellent Talents (No. N130502001), the National Basic Research Program of China (973 Program No. 2014CB643300), the National Environmental Corrosion Platform (NECP) and the “Young Merit Scholars” program of the Institute of Metal Research, Chinese Academy of Sciences.

Li Huabing and Zhou Enze contributed equally to this work.

School of Metallurgy, Northeastern University, Shenyang, 110819, China

Huabing Li, Enze Zhou, Hao Feng & Zhouhua Jiang

Corrosion and Protection Center, University of Science and Technology Beijing, Beijing, 100083, P.R. China

Dawei Zhang & Xiaogang Li

Institute of Metal Research, Chinese Academy of Sciences, Shenyang, 110016, China

Dake Xu, Chunguang Yang & Ke Yang

College of Chemistry, Liaoning University, Shenyang, 110036, China

Jin Xia

Department of Chemical and Biomolecular Engineering, Institute for Corrosion and Multiphase Technology, Ohio University, 45701, Athens, Ohio, USA

Tingyue Gu

You can also search for this author in PubMed Google Scholar

H.L., E.Z. and D.X. designed the experiments and conducted the data analysis; H.L., E.Z., C.Y., D.X., J.X. and H.F. carried out the experiments; H.L., E.Z., D.Z., D.X., Z.J. and K.Y. contributed to the planning and coordination of the project; H.L., E.Z., X.L., D.X. and T.G. wrote and edited the manuscript. All authors participated in discussion about the results and the manuscript.

The authors declare no competing financial interests.

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

Reprints and Permissions

Li, H., Zhou, E., Zhang, D. et al. Microbiologically Influenced Corrosion of 2707 Hyper-Duplex Stainless Steel by Marine Pseudomonas aeruginosa Biofilm. Sci Rep 6, 20190 (2016). https://doi.org/10.1038/srep20190

Download citation

Received: 16 July 2015

Accepted: 23 December 2015

Published: 05 February 2016

DOI: https://doi.org/10.1038/srep20190

Anyone you share the following link with will be able to read this content:

Sorry, a shareable link is not currently available for this article.

Provided by the Springer Nature SharedIt content-sharing initiative

Journal of Bio- and Tribo-Corrosion (2023)

Journal of Polymers and the Environment (2022)

Environmental Science and Pollution Research (2021)

Biotechnology Letters (2020)

Journal of Iron and Steel Research International (2019)

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

What is ERW Steel Pipe?

The Islander Classifieds: Wednesday, July 12, 2023

News

Microbiologically Influenced Corrosion of 2707 Hyper