Portraying manganese biofilms via a merger of EPR spectroscopy and cathodic polarization.

Microbial biofilms on stainless steel surfaces exposed to water from a freshwater pond were dominated by manganese-oxidizing bacteria, as initially diagnosed by microscopy and elemental analysis. The application of electron paramagnetic resonance (EPR) spectroscopy revealed conspicuous sextet (six-line) patterns that intensified with immersion time, implying the gradual accumulation of Mn(II) in the biofilms. Correspondingly, cathodic polarization designated the manganese oxide (MnOx) reduction peak in the form of a distinctive 'nose', which grew increasingly more negative with biofilm growth. The progressive expansion of cathodic current densities and the concurrent area-under-the-curve also allowed the quantification of microbially mediated MnOx deposition. Furthermore, the merger of EPR and cathodic polarization techniques yielded key insights, in tandem with Mn speciation data, into the pathways of microbial manganese transformations in biofilms, besides providing meaningful interpretations of prevailing literature. Accordingly, the natural freshwater biofilm was inferred as one supporting a complete manganese cycle encompassing multiple redox states.

The relationship between alloying elements and biologically produced ennoblement in natural waters.

A range of stainless steels, nickel-chromium and nickel-chromium-molybdenum alloys were exposed to coastal seawater from Mandapam (Indian Ocean) and freshwater from a perennial pond. Biofilms from both test waters produced an ennoblement of the open circuit potential (OCP) on all alloys as expected, which was slower but substantially larger in freshwater. In both waters an interesting relationship was perceived between the plateau OCP (Emax) and the mass percentage of the major alloying elements. In particular, iron exhibited strong positive correlations with Emax (r(2) ≥ 0.77; p < 0.0005), while the sum of chromium, nickel and molybdenum presented significant negative correlations (r(2) ≤ -0.81; p = 0.0002). Consistent with the regression analyses, Euclidean distance clustering yielded patterns where Inconel-600 and the nickel-chromium-molybdenum alloys had the smallest similarities of OCP with other alloys. The results emphatically reinforce a key role for surface passive films in the ennoblement phenomenon in natural waters.

The enrichment of surface passive film on stainless steel during biofilm development in coastal seawater.

The surface passive film on UNS S30400 alloy was characterized before and after biofilm development under different regimes of diurnal lighting in quiescent flowing coastal seawater. As exemplified by atomic force microscopy, the passive film grew under all test conditions with conspicuous variations in morphological features. X-ray photon spectroscopy illustrated an enrichment of the outer film by iron oxide and a progressive increase in the iron oxide/chromium oxide ratio with lighting. Mott-Schottky plots reflected the duplex nature of the film, comprising an outer n-type and an inner p-type configuration. The slopes of the plots showed a strong decrease in donor and acceptor densities with biofilm coverage and lighting, thus confirming passive film growth. These results provide new insights that passive film enrichment is an intrinsic process under practical marine conditions, and show that the evolution of the passive film is a key step to sustained passivity and/or its breakdown by microbial mechanisms.

Stainless steel in coastal seawater: sunlight counteracts biologically enhanced cathodic kinetics.

The influence of sunlight of varying intensity on the performance of UNS S30400 stainless steel (SS) was explored under conditions of natural biofilm development in coastal seawater. In a series of tests performed outdoors under an opaque roof, a range of shades were fashioned to impart varied amounts of diurnal light. These were an ambient level where the underwater illumination was ~ 5% of full sunlight, two intermediate ranges of lighting with ~ 2.5% and ~ 1% of the daylight, and a condition of full darkness. In comparison with the dark, increments of sunlight rendered the SS progressively less aggressive as cathodes in galvanic couples with UNS C70600 alloy. Likewise, welded SS with pre-initiated localized corrosion sites exhibited substantially lower rates of propagation with light. Thus, biofilms and sunlight affected cathodic kinetics in opposite ways. Surface analytical tests showed that the accumulation of manganese (Mn) within the biofilms was small relative to reports from waters of lower salinity. These results not only reveal that extremely low amounts of sunlight are adequate to offset the microbial effect, but also highlight the lack of convincing evidence for Mn cycling as a potent mechanism for enhanced cathodic kinetics in full-strength seawater.

Sunlight-enhanced calcareous deposition on cathodic stainless steel in natural seawater.

In replicate series of experiments in natural seawater, one in full darkness and the other in a 1:1 diurnal cycle with as little as ~5% of natural solar illumination, sunlight promoted calcareous deposition on cathodic stainless steel surfaces. As exemplified by scanning electron microscopy, the deposit that formed under the natural diurnal cycle, in the presence of photosynthetic biofilms, was composed of finer calcareous crystals that provided more compact and more uniform surface coverage than the one formed in the dark. The light-enhanced deposit also possessed better scale properties, as suggested by X-ray analysis and electrochemical measurements. Sunlight enhancement of calcareous deposition looked all the more conspicuous when day and night regimes were examined independently. These results not only bear important implications for cathodic protection in marine waters, but also provide an intriguing analogy to coral reef calcification.

Corrosion behaviour and biofouling characteristics of structural steel in the coastal waters of the Gulf of Mannar (Bay of Bengal), India.

The corrosion behaviour and biofouling characteristics of structural steel coupons at three different locations in the Gulf of Mannar were studied over a period of 2 years. Oyster fouling was predominant at Tuticorin open sea, while barnacle fouling was more pronounced at Mandapam and Tuticorin harbour. Among the three locations, Tuticorin open sea showed a markedly higher biomass, particularly after 12 and 18 months. The extent of crevice corrosion caused by hard foulers was more pronounced at Tuticorin harbour when compared to that at the other two locations. The corrosion rate of the structural steel coupons for 24 months was in the order, Mandapam > Tuticorin harbor > Tuticorin open sea. The loss in tensile strength at 12 and 24 months was in the order, Tuticorin open sea > Tuticorin harbor > Mandapam. The corrosion behaviour of the structural steel coupons was strongly influenced by the variations in the biofouling assemblage at the three different coastal locations.

The influence of sunlight on the localized corrosion of UNS S31600 in natural seawater.

Tests were conducted on the performance of UNS S31600 stainless steel (SS) in a natural day/night cycle vs full darkness under conditions of natural marine biofilm accumulation. In quiescent flowing seawater tests in the laboratory as well as under natural immersion in the sea, diffuse sunlight (∼10% of natural) counteracted the influence of marine biofilms and produced substantial inhibition of the corrosion of SS. Thus, the probabilities (percentage attack) and propagation rates (depths of attack) in multiple crevice tests were substantially lower in the day/night cycle than in the dark. A benefit was also observed for welded SS in terms of the time to corrosion initiation and the mass loss. SS in the passive state showed broader passive regions, well-defined breakdown potentials and markedly smaller anodic and cathodic current densities under the diurnal cycle. The overall reduction in corrosion is attributed to a combination of electrochemical photoinhibition and simultaneous photoinactivation of microbially mediated metal redox reactions linked to cathodic kinetics. These data offer fresh insights into the behaviour of SS under practical seawater situations and the proposed potential use of illumination in the mitigation of biologically influenced consequences.

Cathodic polarization enables SEM illustration of manganese biomineralization in natural biofilms.

Manganese biomineralization occurred readily on metal surfaces in a natural freshwater environment. SEM demonstration of actual bacterial participation was challenging due to the rapidity of biofilm growth and typical microbe-mineral agglomeration. By optimizing cathodic polarization, we slowed down the process so effectually as to document biomineralization by rod-shaped rather than by filamentous bacteria which occurred more frequently at open circuit.

Cathodic behaviour of stainless steel in coastal Indian seawater: calcareous deposits overwhelm biofilms.

Type-316 stainless steel (SS) was investigated as the cathode in galvanic couples in full-strength seawater from the Gulf of Mannar on the southeast coast of India. Tests were devised to examine the impact of SS cathodes on anode materials with or without the accrual of marine biofilms. Biofilmed SS cathodes significantly enhanced the rate of corrosion of nickel, causing noble shifts in the couple potentials. With mild steel and zinc as the anodes, calcareous deposits developed quite rapidly on the SS cathodes and led to a significant reduction of bacterial numbers. The calcareous deposits also caused substantial reduction of galvanic corrosion rates for mild steel, whereas there was no difference for zinc. The deposits were identified by XRD as essentially carbonates, oxides and hydroxides of calcium and magnesium. Potentiodynamic polarization performed on the actual couples after disconnection and equilibration provided reasonable interpretations of the galvanic corrosion trends. Data from this work suggest that a potential of about -0.70 V vs. saturated calomel electrode (SCE) should provide optimum protection of SS in warmer, full-strength seawater that supports the precipitation of calcareous deposits. The criterion commonly recommended for temperate conditions of lower water temperature and estuarine waters of lower alkalinity is -1.0 V (SCE).

On microalgal settlements and the sluggish development of marine biofouling in Port Blair waters, Andamans.

Settlement of microalgae was investigated on Perspex, aluminium and zinc coupons immersed in Port Blair Bay waters for over 3 months. Commencement of fouling was exceptionally slow, and few microalgae were found until 14 days. Settlement occurred thereafter, and 47 microalgal species contributed to the fouling. The dominant forms belonged to the genera Navicula and Nitzschia, whereas Coscinodiscus eccentricus, Gyrosigma balticum and Trichodesmium erythraeum also accounted for high proportions of the settlements. The dominance of Nitzschia sigma was particularly marked on zinc coupons, suggesting an ability by the organism to resist toxicity. Settlement of both centric and pennate diatoms was observed in the early and mid periods, and absolute dominance of the pennate diatoms subsequently. The fouling mass was low even after 103 days, and it is speculated that strong ultraviolet radiation might be the prime reason for the sluggish development of marine biofouling in these oceanic island waters.