Acid mine drainage (AMD) is a worldwide environmental problem associated with the mining wastes, generated from active and inactive mining sites from mineral processing activities. AMD is defined as the drainage that occurs as a result of oxidation of sulphide minerals/wastes/tailings when exposed to air and water in the presence of chemolithotrophs namely the Acidithiobacillus sp. AMD is characterised by low pH and increased acidity due to elevated heavy metals and sulphate concentration. The acid production potential was carried out for sulphide minerals such as pyrite and chalcopyrite and copper tailings sample in the absence and presence of bacteria namely Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans. Acidity was generated in all the cases due to the oxidation of the mineral samples. The oxidation was found to occur at a higher rate in the presence of the bacteria compared to the control samples. Bioremediation experiments were conducted on acid mine water collected from the Ingaldahl Mines, Chitradurga, Karnataka, India, using organic and inorganic substrates. In the experiments with rice husk, complete removal of metal ions from the acid mine water was achieved with an attendant increase in the pH of the acid mine water from 2.3 to 5.5. About 21% of sulphate could be removed using Dsm. nigrificans from acid mine water pretreated with rice husk at pH 5.5 and this was further increased to 40% by the supplementation of organic components. The rice husk filtrate was found to serve as a good growth medium for Dsm. nigrificans. About 96 % of Fe, 75 % of Zn, 92 % of Cu and 41 % of sulphate removal was achieved from the acid mine water of pH 2.4 with a concomitant increase in the pH value by two units after interaction with the tree bark. About 56 % and 71 % of sulphate reduction could be achieved at initial pH values of 4.1 and 5.5 respectively of the acid mine water pretreated with E. tereticornis (Sm) bark, after inoculation with Dsm. nigrificans. The complete removal of Fe2+ and Fe3+, 80% of Zn, 83% of Cu and 62% of sulphate could be removed from acid mine water using fly ash as the substrate with an increase in pH of acid mine water from 2.3 to 7. About 68% of sulphate reduction at pH 6.8 could be achieved in acid mine water pretreated with fly ash in the presence of Dsm. nigrificans. With red mud as the substrate, complete removal of all the metal ions namely Fe2+, Fe3+, Zn, and Cu from acid mine water was achieved with a concomitant increase in the pH from 2.3 to 8. The sulphate reduction was increased to about 51% at pH 7.2 when the acid mine water pretreated with red mud was inoculated with Dsm. nigrificans. The adsorption experiments carried out on the acid mine water using either organic or inorganic substrates indicated that the free energy of adsorption was negative for all the chosen metal ions attesting to favorable interaction. The adsorption isotherms of the metal ions for rice husk exhibited Langmuirian behaviour, while those for the other substrates adhered to both Langmuir and Freundlich relationships. The adsorption process was found to be endothermic in nature for rice husk, fly ash and red mud. On the contrary, the adsorption onto tree bark showed exothermic behaviour. The adsorption kinetics of the metal ions onto the various substrates adhered to the first order Lagergren equation. The metal uptake processes by the organic and inorganic substrates chosen for this study involve ionic, chemical and physical forces of adsorption. The different types of functional groups present on the surface of the substrates such as carboxyl, hydroxyl and carbonyl, as revealed by FTIR spectroscopic studies, partake in metal binding. The metal ions will also be adsorbed by complexing with the negatively charged reaction sites on the substrate surfaces. Furthermore, the complex solution chemistry of the metals as a function of pH has also to be taken into consideration. The mechanism of sulphate reduction by Dsm. nigrificans in the presence of organic carbon can be illustrated as: 2CH2O + SO42- + 2H+  2CO2 + 2H2O + H2S M2 + H2S  MS  + 2H+ where, CH2O represents the organic matter and M represents the metal ion.

[pt] Este trabalho teve por objetivo avaliar a morfologia de biofilmes, formados em superfície de aço API 5L X80 submetida a um fluido com baixo BSW (Basic Sediment Water), ou seja, baixo teor de água em óleo, através de análise por Microscopia Eletrônica de Varredura (MEV), determinar a cinética e quantificar os grupos bacterianos de formação destes biofilmes, e avaliar a biocorrosão causada por estes microrganismos. Foram realizadas alterações no protocolo original de preparo de amostras, com a finalidade de obter melhor definição das morfologias avaliadas. Realizaram-se estudos dinâmicos (looping) e em condições estáticas, com e sem a adição de agente biocida. Utilizando-se como fluido de processo o óleo coletado no terminal da Petrobras em Barra do Furado que tem como característica um BSW de aproximadamente 1 por cento de água. Para alcançar os objetivos descritos foram quantificadas bactérias sésseis (bactérias facultativas heterotróficas, bactérias precipitantes do ferro, bactérias anaeróbias heterotróficas e bactérias redutoras de sulfato) através da técnica do número mais provável (NMP). A intensidade do processo corrosivo foi avaliada através da análise dos pites formados nos cupons por microscopia óptica. A seqüência de fixação com alteração do protocolo inicial, permitiu uma melhor preservação estrutural do biofilme obtendo-se assim imagens com melhor resolução, tendo em vista que a fixação primária com glutaraldeído fixa principalmente proteínas pelo estabelecimento de ligações divalentes com grupamento amino, e a pós-fixação em tetróxido de ósmio reage com os lipídios. Quanto a cinética, observou-se também que na presença de fluido oleoso, a formação do biofilme só é detectada após 15 dias de exposição. A adição do agente biocida inibiu o crescimento do biofilme, não sendo detectada a presença de células cultiváveis após 7 dias em contato com o fluido contendo o agente químico. / [en] This study aims to evaluate the morphology of biofilms formed on the surface of API 5L X80 steel exposed to a low BSW (Basic Sediment Water) fluid comprising oil of a low water content. The analisys was performed using scanning electron microscopy (SEM) in order to determine the formation kinetics, to quantify and identify the bacterial groups present as the film formed, and to evaluate the biocorrosion caused by these organisms. Modifications were made to the original sample preparation protocol, in order to better define the film morphologies. Studies were undertaken applying either static or dynamic (looping) conditions, bith with and without the addition of biocides. The process fluid used was oil collected from the Petrobras Barra do Furado Terminal, characterized by its 1 percent BSW. The evaluation was performed quantifying the sessile bacteria present in the film (heterotrophic bacteria, iron bacteria, anaerobic bacteria e sulphate reducing bacteria) via the Most Probable Number (MPN) technique. The intensity of the corrosion process was determined by the analysis of pits forme in the testpieces, by optical microscopy. The application of a sample fixation sequence, in addition to modifications of the inicial sample preparation protocol, permitted the improved preservation of the biofilm structures, thereby permitting higher resolution images. The primary fixing agent, containing gluteraldehyde, principally fixes proteins, by establishing divalent bonds with amino groups, while the post-fixation stage with osmium tetroxide invloves reactions with lipid groups.With regard to the film deposition kinetics, it was observed that biofilm formation could only be detected after 15 day s exposure to the oily fluid. The addition of a biocide, inhibited the growth of the film, no cultivatable cells being present after 7 days contact with the treated fluid.

[pt] A Corrosão Influenciada por Microrganismos (CIM) ou biocorrosão é reconhecida como um dos fenômenos causadores de inúmeros problemas nas indústrias de petróleo e gás, pois causa sérios danos ao material reduzindo sua vida útil. Neste trabalho foram avaliados os efeitos da biocorrosão caracterizado pela cinética de formação de biofilme em uma junta soldada longitudinal de aço API X80 obtida pelo processo de arco submerso (SAW). Durante o processo de soldagem forma-se uma região com características microestruturais distintas do metal de base e do metal de adição, denominada de zona termicamente afetada (ZTA). Assim essa zona poderá ter uma adesão microbiana diferenciada, visto que diferenças superficiais em um material, seja por natureza química ou física, podem limitar ou facilitar a adesão microbiológica. Por esse motivo foi realizado um estudo comparativo entre a cinética da formação de biofilme na junta soldada de um aço API X80. Também foi avaliada a influência das características físicas da superfície na adesão microbiana utilizando dois tipos de superfície: com a rugosidade original e polido com pasta de diamante com granulação de 6μm. Estas superfícies foram expostas ao fluido de processo (água do mar da Baía de Guanabara) em um sistema dinâmico. Foram realizadas tanto a quantificação microbiana, como também a quantificação dos ácidos orgânicos, sulfato depletado e ferro total para avaliar os nutrientes disponíveis e a bioatividade das reações bacterianas. A rugosidade superficial e o biofilme formado foram caracterizados morfologicamente e a sua presença correlacionada com a formação de pites. / [en] The Microbiologically Influenced Corrosion (MIC) or bio-corrosion is recognized as one of the phenomena that cause a lot of problems in petroleum and gas industries, because it causes serious damages to the materials, reducing its life cycle. This study evaluated the effects of bio-corrosion characterized by the biofilm formation kinetics in longitudinal welds of API 5L X80 steel obtained by the process of submerged arc welding (SAW). During the welding process a region with different microstructural characteristics of the base metal and weld metal is formed, called heat affected zone (HAZ). Thus this zone can have a differentiated microbial adhesion since the different surfaces of a material, either by chemical and physical nature, can limit or facilitate the microbial adhesion. For this reason a comparative study of biofilm formation kinetic on the welded joint was conducted. The influence of the physical characteristics of the surface in microbial adhesion was also evaluated using two kinds of surface: steel with real roughness and steel polished with diamond paste with grain size of 6 μm. These surfaces were exposed to the process fluid (Guanabara Bay seawater) in a dynamic flow system. The microbial quantification was held. Organic acids, depleted sulfate and total iron were also measured to evaluate the available nutrients bioactivity of bacterial reactions. Surface roughness and biofilm were characterized morphologically and correlated with pitting formation.