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Occurrence of Manganese-Oxidizing Microorganisms and Manganese Deposition During Bio¢Lm Formation on Stainless Steel in a Brackish Surface Water

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Occurrence of Manganese-Oxidizing Microorganisms and Manganese Deposition During Bio¢Lm Formation on Stainless Steel in a Brackish Surface Water FEMS Microbiology Ecology 39 (2002) 41^55 www.fems-microbiology.org Occurrence of manganese-oxidizing microorganisms and manganese deposition during bio¢lm formation on stainless steel in a brackish surface water Jan Kielemoes a, Isabelle Bultinck b, Hedwig Storms b, Nico Boon a, Willy Verstraete a;* Downloaded from https://academic.oup.com/femsec/article/39/1/41/535962 by guest on 29 September 2021 a Laboratory of Microbial Ecology and Technology (LabMET), Faculty of Agricultural and Applied Biological Sciences, Ghent University, Coupure Links 653, B-9000 Ghent, Belgium b OCAS N.V., John Kennedylaan 3, B-9060 Zelzate, Belgium Received 25 May 2001; received in revised form 8 October 2001; accepted 10 October 2001 First published online 5 December 2001 Abstract Biofilm formation on 316L stainless steel was investigated in a pilotscale flow-through system fed with brackish surface water using an alternating flow/stagnation/flow regime. Microbial community analysis by denaturing gradient gel electrophoresis and sequencing revealed the presence of complex microbial ecosystems consisting of, amongst others, Leptothrix-related manganese-oxidizing bacteria in the adjacent water, and sulfur-oxidizing, sulfate-reducing and slime-producing bacteria in the biofilm. Selective plating of the biofilm indicated the presence of high levels of manganese-oxidizing microorganisms, while microscopic and chemical analyses of the biofilm confirmed the presence of filamentous manganese-precipitating microorganisms, most probably Leptothrix species. Strong accumulation of iron and manganese occurred in the biofilm relative to the adjacent water. No evidence of selective colonization of the steel surface or biocorrosion was found over the experimental period. The overall results of this study highlight the potential formation of complex microbial biofilm communities in flow-through systems thriving on minor concentrations of manganese. ß 2002 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved. Keywords: Bio¢lm; Pilotscale simulation; Stainless steel; Manganese-oxidizing microorganism; Biocorrosion 1. Introduction potential of the steel (Ecorr or corrosion potential) in a positive or negative direction, depending on the nature Biocorrosion of stainless steel can be catalyzed by a of the deposit. In principle, the more positive the poten- series of metabolic activities of a wide range of microor- tial, the better metal ions are capable to leave the metal ganisms. In the past, sulfate-reducing bacteria (SRB) were surface. Pitting corrosion of stainless steel occurs when considered to be the most common catalysts of biocorro- this open-circuit potential becomes equal to or more pos- sion, whereas the role of other bacteria has been seriously itive than the pitting potential of the steel (Ep). Manganese neglected [1]. Moreover, aerobic bacteria generally have oxide biodeposition on stainless steel surfaces for example much higher growth rates than anaerobes, and their meta- can force a shift of Ecorr in the positive direction, making bolic activity is much higher, potentially leading to higher some stainless steels more vulnerable to pitting and crevice corrosion rates. Consequently, other microorganisms than corrosion [2,3]. On the other hand, bioprecipitated sul¢des SRB are currently suspected to be related to biocorrosion can lead to a lower potential of the steel. Mineral disso- in industrial systems. Microorganisms performing miner- lution reactions can remove protective passive layers (e.g. alization reactions for instance can in£uence corrosion by Cr and Fe oxide layers in the case of stainless steel) or both forming and dissolving mineral deposits [2]. Mineral force mineral replacement reactions that lead to further deposition on a metal surface can shift the open-circuit metal dissolution: e.g. the metal-reducing bacterium She- wanella putrefaciens can reduce solid Fe3 oxide to soluble Fe2 oxide [4]. According to the mixed potential theory * Corresponding author. Tel.: +32 9 264-59-76; Fax: +32 9 264-62-48. this cathodic reaction consumes electrons produced by the E-mail address: [email protected] (W. Verstraete). anodic dissolution of the underlying metal, thus increasing 0168-6496 / 02 / $22.00 ß 2002 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved. PII: S0168-6496(01)00191-X FEMSEC 1307 26-2-02 Cyaan Magenta Geel Zwart 42 J. Kielemoes et al. / FEMS Microbiology Ecology 39 (2002) 41^55 the corrosion rate. Interrelationships and cooperation be- were used in the experiments. The £ow/stagnation/£ow tween for instance manganese- and iron-oxidizing bacteria regime was used to simulate the hydrostatic testing prac- and SRB can lead to a more severe corrosion than can be tice. Water systems are subjected to a hydrotest before expected from each individual member of such a complex putting them into service. At least one failure occurred microbial community. A model for these interactions was after a hydrotest leaving the brackish surface water in already proposed by Dickinson and Lewandowski [5]: the piping system for several weeks before the start-up. MnO2 depolarization shifts Ecorr to values exceeding Ep The bio¢lm was extensively characterized and the e¡ect at local sites of diminished redox potential (SRB niche). of bio¢lm development and its reaction products on the In laboratory bio¢lm and/or corrosion experiments pure corrosion of the stainless steel was investigated. cultures are often used to obtain a better understanding of the mechanisms of bacterial adhesion, bio¢lm formation and corrosion in more complex industrial systems. The 2. Materials and methods dynamic interactions of di¡erent organisms together with Downloaded from https://academic.oup.com/femsec/article/39/1/41/535962 by guest on 29 September 2021 a constantly changing environment as in the case of nat- 2.1. Pilotscale £ow-through system ural waters cannot be simulated in laboratory experiments only. The processes of bio¢lm formation and corrosion are Fig. 1 represents the design of the pilotscale £ow- interrelated and observed corrosion rates in practice rarely through system. Five transparent, disconnectable PVC correlated with pure-culture simulations. Detailed back- tubes (inner diameter = 36 mm and length = 1 m) were ground information and the application of the appropriate coupled to a water reservoir of about 0.6 m3 and mounted analytical techniques to determine this multiplicity of pa- in a closed wooden construction to control the develop- rameters are vital in corrosion failure analyses or during ment of photosynthetic bacteria and algae. Stainless steel simulation experiments in order to reach reliable conclu- coupons (2.9U6.9 cm) were tungsten inert gas welded sions. grade AISI 316L sheets (2 mm thick) with a smooth 2B The aim of the present work was to study in situ natural surface ¢nish obtained from stainless steel producer ALZ bio¢lm adhesion and biocorrosion simulation experiments (Genk, Belgium). Five welded stainless steel coupons to- on welded stainless steels in a brackish surface water using gether with one microscope glass slide as reference materi- a pilotscale £ow-through system with an alternating £ow/ al were mounted in Ertalon1 rails with the weld perpen- stagnation/£ow regime. In the past, the application of this dicular to the £ow direction, electrically insulated from brackish surface water in cooling water systems resulted in one another to prevent galvanic e¡ects. The £ow-through numerous corrosion failures of type 316L stainless steel medium was natural brackish surface water (canal water tubes, some of which have been tentatively attributed to Ghent^Terneuzen, Belgium), indicated further as `canal microbial in£uences. As the corrosion was predominantly water'. The chemical composition of the water in the res- appearing at the circumferential unpickled welds, welded ervoir of the pilot plant system (sample point D, Fig. 1) stainless steel coupons without removal of the heat tints measured at the end of the experiment (representative for Fig. 1. Design of the pilotscale £ow-through installation. Sampling points: (A) fresh canal water, (B) water in the reservoir after manually mixing and stirring the content, (C) sedimented sludge at the bottom of the tank, (D) water at the upper part of the reservoir (mixing zone of incoming fresh canal water and the water present in the tank) and (E) water at the end of the over£ow tubing. FEMSEC 1307 26-2-02 Cyaan Magenta Geel Zwart J. Kielemoes et al. / FEMS Microbiology Ecology 39 (2002) 41^55 43 Table 1 chloride (CTC)/4,6-diamidino-2-phenylindole (DAPI) (see Chemical composition of the water in the reservoir of the pilot plant below). Procedure B was intended to analyze the chemical system (sample point D, Fig. 1) measured at the end of the experiment composition of the adhered material. The bio¢lm at both (expressed in mg l31, except pH) sides of the coupon, except from the welded area and the Parameter Value Parameter Value heat-a¡ected zone (HAZ), was removed by swabbing with pH 7.7 Kj-N 5.0 cotton wool sticks and transferred into 30-ml sterile Milli- Suspended solids 3 NO3-N+NO3-N 6.5 3 2 Q water. These solutions were vortexed intensively during Chemical oxygen demand 24 Cl3 453 5 min to break up particulate matter and some important Biological oxygen demand 6 Natotal 148 Dry residue 1550 Ktotal 17 chemical parameters were determined (see below). Proce- Ash residue 1180 Mgtotal 25 dure C was almost identical to procedure B, with the ex- SO23-S 55 Ca 133 4 filtrate total ception that the coupons were swabbed completely at both Ptotal 0.3 Fetotal 1.70 sides. This is done in order to evaluate the e¡ect of weld- NH-N 0 Mn 0.23 4 total ing on the chemical/biological composition of the bio¢lm. Downloaded from https://academic.oup.com/femsec/article/39/1/41/535962 by guest on 29 September 2021 In procedure D one side of the coupons was sampled ac- cording to procedure C.
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