The development of biofilms on mild and stainless steel surfaces in pure and mixed batch cultures of the bacterial species Pseudomonas fluorescens and Desulfovibrio desulfuricans and the role of these biofilms in corrosion of steel has been investigated. Early events leading to the formation of biofilms have been elucidated by studying the attachment of bacterial cells to steel using epifluorescence microscopy. To identify the nature of the bacterial surface components involved in the initial adhesion to mild steel, lectins, their sugar inhibitors and saccharolytic and proteolytic enzymes have been employed. Polyclonal antibodies have been raised against bacteriallipopolysaccharides (LPS) and their influence on bacterial adhesion assessed. LPS have been analysed chemically by gas-chromatography (GC-FID) and gas chromatography-mass spectrometry (GC-MS) to determine their ~arbohydrate composition and fatty acid content. On the basis of the results obtained the ~nvolv~~~nt of glucose and N-acetylglucosamine, present in O-antigenic fractions of LPS, 10 the lOlnal attachment of the two bacterial species to mild steel is suggested. Both types of carbohydrates are likely to be involved in early attachment of Pseudomonas to mild steel, whereas only a polymeric fonn of N-acetylglucosamine seems to participate in adhesion of Desulfovibrio. The subsequent biofilm development on steel surfaces and their accompanying corrosion h~s been monitored by scanning electron microscopy (SEM). SEM studies reveal very different patterns of bacterial biofilms on mild and stainless steel and show varied degrees of corrosion occurring on these surfaces. Thin and patchy Pseudomonas biofilms are accompanied by little corrosion whilst thick. more continuous, Desul/ovibrio biofilms are associated with higher levels of corrosion. Energy dispersive X-ray analysis (BOAX) of corrosion products present on steel surfaces indicates ferrous sulphides as the major components in Desul/ovibrio biofilms. The corrosion of steel in bacterial cultures has also been investigated by kinetic polarisation measurements. The results obtained from cathodic and anodic polarisation curves, combined with SEM and EDAX analyses confmn the SEM observation. Stainless steel is not subjected to any great degree of fouling or corrosion under the chosen experimental conditions. The EPS associated with biofilms and released into the liquid phase of the culture media (free EPS) has been characterised. Proteins and carbohydrates in these polymers are detected colorimenically and by SDS-gel electrophoresis. Uronic acids, found in biofilmbound BPS. are not detected in free EPS. The GC-MS and GC-FIO analyses have aided in establishing types and quantities of neutral carbohydrates present in bacterial exopolymers and show that the neutral sugar composition of free and surface-associated BPS is not identical for a given bacterial culture. The biofilm-bound BPS are believed not to play a major role in corrosion of mild steel but to provide additional mechanisms in its facilitation. No correlation between levels of free BPS and corrosion of steel is found.

No description available.

Materiais metálicos expostos a ambientes aquáticos estão susceptíveis ao desenvolvimento de Corrosão Microbiologicamente Influenciada (CMI). Nas indústrias a CMI é um assunto de extrema importância devido aos prejuízos gerados nos reparos ou trocas de equipamentos e nas interrupções dos sistemas de produção associadas às falhas técnicas. No presente trabalho estão apresentados resultados de um estudo de CMI em superfícies metálicas expostas à água de processo em uma unidade geradora de energia hidrelétrica, que sofre com um acelerado processo corrosivo, principalmente nos trocadores de calor e nas turbinas. A partir de biofilmes de corrosão amostrados nas turbinas e trocadores de calor foram isolados grupos bacterianos redutores de sulfato e oxidantes e redutores de ferro que foram relacionadas como indutoras/ agravantes do processo corrosivo. / Metallic materials exposed to aquatic environments are susceptible to development of Microbiologically Influenced Corrosion (MIC). In industries MIC is a matter of great importance due to damages generated in repairs or equipment changes and production systems disruptions associated with technical failures. In this work are presented results of a MIC study on metal surfaces exposed to process water in a generating unit of hydropower, which suffers from an accelerated corrosion process, especially in the heat exchangers and the turbines. From corrosion biofilms sampled in turbines and heat exchangers were isolated bacterial groups sulfate reducing and oxidizing and reducing iron. These bacteria have been widely reported in the literature as inducing / aggravating the corrosion process.

Materiais metálicos expostos a ambientes aquáticos estão susceptíveis ao desenvolvimento de Corrosão Microbiologicamente Influenciada (CMI). Nas indústrias a CMI é um assunto de extrema importância devido aos prejuízos gerados nos reparos ou trocas de equipamentos e nas interrupções dos sistemas de produção associadas às falhas técnicas. No presente trabalho estão apresentados resultados de um estudo de CMI em superfícies metálicas expostas à água de processo em uma unidade geradora de energia hidrelétrica, que sofre com um acelerado processo corrosivo, principalmente nos trocadores de calor e nas turbinas. A partir de biofilmes de corrosão amostrados nas turbinas e trocadores de calor foram isolados grupos bacterianos redutores de sulfato e oxidantes e redutores de ferro que foram relacionadas como indutoras/ agravantes do processo corrosivo. / Metallic materials exposed to aquatic environments are susceptible to development of Microbiologically Influenced Corrosion (MIC). In industries MIC is a matter of great importance due to damages generated in repairs or equipment changes and production systems disruptions associated with technical failures. In this work are presented results of a MIC study on metal surfaces exposed to process water in a generating unit of hydropower, which suffers from an accelerated corrosion process, especially in the heat exchangers and the turbines. From corrosion biofilms sampled in turbines and heat exchangers were isolated bacterial groups sulfate reducing and oxidizing and reducing iron. These bacteria have been widely reported in the literature as inducing / aggravating the corrosion process.

La corrosion induite par les micro-organismes (CIM) génère des pertes économiques mondiales chiffrées en milliards d’euros par an. Il est communément admis que les bactéries sulfato-réductrices (BSR) jouent un rôle clé dans la CIM anaérobie des aciers. Malgré cette unanimité, les essais en laboratoire peinent à reproduire la corrosion des aciers observées en milieu naturel; bien plus, ils n’expliquent pas quel est l’élément qui déclenche la corrosion, puisque les BSR présentes dans de nombreux environnements naturels n’induisent pas systématiquement de corrosion. L’objectif de ce travail est d’évaluer la pertinence dans le domaine de la CIM de nouveaux mécanismes de transferts électroniques entre aciers et protéines ou cellules microbiennes. La première partie de la thèse évalue l’effet d’une [Fe]-hydrogénase sur les processus de corrosion anaérobie des aciers au carbone. L’hypothèse d’une catalyse directe de la réduction des protons par des hydrogénases adsorbées a souvent été suggérée dans la bibliographie, elle est ici clairement démontrée. L’hydrogénase de Clostridium acetobutylicum, qu’elle soit active, désactivée ou dénaturée accélère la corrosion de l’acier au carbone. La présence de phosphate dans le milieu rend les interprétations plus complexes mais ne modifie pas le mécanisme. Une nouvelle hypothèse est avancée qui donne un rôle essentiel aux centres fer-soufre de la protéine. La catalyse de la corrosion par les hydrogénases pourrait donc être rapprochée des mécanismes bien connus de catalyse par le sulfure de fer. Dans ce cas l’état redox des centres fer-soufre serait une clé essentielle de l’apparition ou non de la corrosion. La deuxième partie élucide le rôle de Geobacter sulfurreducens sur la corrosion anaérobie de trois types de matériaux : aciers au carbone (1145), ferritique (403) et austénitiques (304L et 316L). Les résultats mettent en évidence pour la première fois que des cellules bactériennes adhérées induisent un anoblissement du potentiel libre des aciers et accélèrent la corrosion des aciers faiblement alliés par un mécanisme de transfert direct d’électrons. Suivant les concentrations d’accepteurs et de donneurs d’électrons en solution, G. sulfurreducens peut accentuer la propagation de la corrosion en catalysant directement la réduction cathodique ou, au contraire, en absence d’accepteurs et en excès de donneurs, protéger contre la corrosion. L’apparition de la corrosion ne peut donc être induite que par la conjonction défavorable de plusieurs paramètres. Ces résultats obtenus en laboratoire apportent de nouvelles voies d’investigations des phénomènes de CIM qui doivent maintenant être confrontées aux milieux naturels. / Microbially influenced corrosion (MIC) costs billions of euros per year. It is commonly agreed that sulphate-reducing bacteria (SRB) play a key role in anaerobic MIC of steels. In spite of this, laboratory experiments have difficulty in reproducing the corrosion of steels that is observed in natural environments. Moreover, they do not explain what triggers corrosion since SRB, ubiquitous in natural environments, do not systematically induce corrosion. The aim of this work was to evaluate the relevance of new electron transfer mechanisms between steels and proteins or microbial cells in the domain of MIC. The first part of the thesis evaluates the impact of [Fe]-hydrogenase on the anaerobic corrosion of mild steels. The direct catalysis of proton reduction by hydrogenases has often been suggested in the literature; here, it is clearly demonstrated. Hydrogenase from Clostridium acetobutylicum, whether it is active, deactivated on denatured, can accelerate the corrosion of mild steel. The presence of a phosphate medium makes the interpretations more complex without modifying the mechanism. A new hypothesis implying the crucial role of iron-sulphur clusters contained in the protein is brought to light. Corrosion catalysis by hydrogenases could be compared with well-known mechanisms of corrosion catalysis by iron sulphide. In this case, the redox state of iron-sulphur clusters would play a key role in the occurrence of corrosion. The second part elucidates the role of Geobacter sulfurreducens in anaerobic corrosion of three types of steels: mild steel (1145), ferritic (403) and austenitic steels (304L and 316L). Results show, for the first time, that adherent bacterial cells induce open circuit potential ennoblement of steels and accelerate the corrosion of slightly alloyed steels by a direct electron transfer mechanism. Depending on the concentrations of the electron acceptors and donors in the medium, G. sulfurreducens could either enhance corrosion propagation by direct catalysis of proton reduction or, in the absence of acceptors and with an excess of donors, protect against corrosion. Thus the occurrence of corrosion relies on the unfavourable conjunction of many parameters. These results obtained in laboratory conditions open new paths for investigating MIC in natural environments.

No description available.