A Drenagem ácida de minas (DAM), do ponto de vista ecológico, influencia os ambientes aquáticos provocando condições de stress para a maioria dos organismos. O baixo valor de pH (geralmente < 3,0), altas quantidades de sulfatos e íons metálicos dissolvidos diminuem a diversidade biológica. Esses locais podem ser caracterizados como ecossistemas simples, dominados por organismos acidofílicos e ácido tolerantes. Assim, o objetivo do presente trabalho foi identificar as comunidades de algas encontradas em ecossistemas impactados pela DAM na região carbonífera do Estado de Santa Catarina, Brasil. Observou-se 12 córregos, dois quais 6 com presença de algas e 6 com ausência de algas. As amostragens foram realizadas em julho/2009, outubro/2009, dezembro/2009 e abril/2010, de forma a abranger as quatro estações do ano. Verificouse que os ambientes aquáticos atingidos pela DAM abrigam pouca diversidade de espécies. A comunidade de algas foi representada por 5 táxons e a identificação realizou-se somente até o nível de gênero. Os gêneros encontrados foram: Microspora, Mougeotia, Eunotia, Frustulia e Euglena. Os gêneros de maior distribuição foram a Microspora, estando presente em 6 locais de amostragem e o gênero Eunotia, presente em 4 locais de amostragem. O gênero em maior abundância e quantidade de biomassa foi a Microspora. O crescimento das algas foi dependente de vários fatores, entre os quais características químicas da água e o período de amostrado. Observou-se que as algas acidofílicas cresceram em corpos d´água na faixa de pH entre 3,2 e 4,1, sendo que nos locais com pH inferior a 2,8 ou superior a 4,2 não ocorreu o crescimento de algas. O gênero Microspora esteve presente em águas com níveis variados de metais e sulfato. Já o gênero Frustulia somente se desenvolveu em corpos d´água com baixos níveis de ferro e sulfato. Por sua vez, o gênero Euglena só foi bem sucedido em locais onde os teores de sulfatos e ferro foram elevados. Sendo assim, as algas encontradas neste estudo apresentam a capacidade de bioindicadoras da qualidade do local. Em relação aos períodos amostrados, a diversidade e abundância de algas foram maiores no inverno. As comunidades perifíticas são capazes de acumular elementos metálicos em sua biomassa. A análise da biomassa perifítica indica que o material contém uma elevada concentração de elementos inorgânicos, bioacumulados do ambiente de crescimento. / The acid mine drainage (AMD), from the ecological point of view, affects the aquatic environments causing stress to most organisms. The low pH values (usually <3.0), high concentration of sulfates and dissolved metal ions reduce the biological diversity. These systems can be characterized as simple ecosystems, dominated by acidophilic and acid tolerant organisms. The objective of this study was to identify communities of algae found in ecosystems impacted by AMD in the coal region of Santa Catarina, Brazil. It was observed 12 streams, 6 of which with the presence of algae and 6 with the absence of algae. Samples were collected in July/2009, October/2009, December/2009 and April/2010 in order to cover all four seasons. It was found that aquatic environments affected by DAM refuge low species diversity. The algal community was represented by 5 taxa and the identification was made only to the genus level. The species identified were: Microspora, Mougeotia, Eunotia, Frustulia and Euglena. The genus of the largest distribution was Microspora, present in 6 sampling sites and Eunotia, present in 4 sampling sites. The genus in greater abundance and amount of biomass was the Microspora. The algal growth was dependent of several factors, including water chemistry and the period of sampling. It was observed that acidophilic algae grown in water with the pH range between 3.2 and 4.1, and in places with a pH below 2.8 or above 4.2 there was no algae growth. The genus Microspora was present in waters with different levels of metals and sulfate. But the genus Frustulia only was increased in water with low levels of iron and sulfate. On the other hand, the genus Euglena has only been successful in places where the levels of sulfates and iron were high. Therefore, the algae found in this study have the ability to bioindicators of the water quality. Concerning the periods sampled, diversity and abundance of algae were higher in winter. Periphyton communities are able to accumulate metallic elements in their biomass. The analysis of periphytic biomass indicates that the material contains a high concentration of inorganic elements, bioaccumulated of the growth environment.

Efluentes contaminados por lixiviados de resíduos sólidos urbanos (LRSU), proveniente de aterros sanitários, ou a drenagem ácida de minas (DAM), originados pela oxidação da pirita em rejeitos de carvão, são problemas atuais em termos de impacto ambiental, especialmente na região do Município de Butiá, no Estado do Rio Grande do Sul. O tratamento destes efluentes, via de regra, é bastante oneroso, mas necessário, uma vez que a liberação in natura destas fontes poluidoras pode ser responsável pela degradação severa do ambiente. Em geral, metodologias convencionais de tratamento (como a precipitação/sedimentação, no caso da DAM, e processos biológicos, no caso do LRSU) mostram-se economicamente custosas ou tecnicamente deficientes. Frente a este desafio, esta pesquisa baseou-se na avaliação do tratamento conjunto de DAM e LRSU, via homogeneização, a fim de diminuir custos tradicionais decorrentes. O processo consiste no aproveitamento da alcalinidade do LRSU para neutralizar a acidez da DAM. Ao mesmo tempo, o ferro e o alumínio presentes na DAM permitem a coagulação dos sólidos suspensos, removendo uma significativa carga de poluentes. Resultados obtidos em laboratório no tratamento inicial destes efluentes permitiram uma significativa redução de carga orgânica, nutrientes, metais pesados e bactérias do grupo coliforme. O efluente final apresentou redução na carga lançada de 100% para sólidos suspensos, 36% para DBO, 56% para DQO, 11% para N, 72% para Fe, 73% para Al e 9% para Mn, atenuando suas características iniciais em consonância a alguns padrões de emissão de efluentes líquidos estabelecidos para o Estado do Rio Grande do Sul (Resolução número 128/2006 do CONSEMA). / Municipal landfills wastewaters (MLW) and acid mine drainages (AMD) from coal mines are environmental issues faced, in special case at the city of Butia, in Rio Grande do Sul State. The treatment of such effluents is necessary to avoid undesirable environmental degradation. The conventional treatment techniques (e.g. chemical neutralization for DAM and biological processes for municipal landfills wastewaters) are costly and sometimes inefficient. This work considered a new alternative based in the combined treatment of MLW and AMD. The alkalinity of MLW can neutralize the acidity of the AMD while the iron and aluminum present in the AMD allow the coagulations of the suspended solids of the MLW, reducing considerably the amount of pollutants. The initial results obtained at laboratory scale allowed the reduction of organic load, nutrients, heavy metals and bacteria. The final effluent showed reduction at the charge reaching 100% to solids in suspension, 36% to BOD, 56% to COD, 11% to N, 72% to Fe, 73% to Al e 9% to Mn, without addiction of coagulations polymers, and improving its characteristics considering the emission standards established in the Rio Grande do Sul State, Brazil (Resolution number 128/2006 from CONSEMA).

A drenagem ácida de minas (DAM) é formada quando certos minerais sulfetados são expostos a condições de oxidação. A DAM é normalmente associada com a mineração de carvão, principalmente de depósitos de rejeitos de carvão, que possuem alta concentração de pirita (FeS2). A neutralização da DAM usando materiais alcalinos, como calcário e escórias de aciaria é uma das técnicas utilizadas para remediação deste problema. Assim, o objetivo deste trabalho foi estudar o desempenho da escória de aciaria elétrica do forno-panela na neutralização da DAM em leitos de percolação em comparação com calcário, material tradicionalmente empregado para tal fim. A metodologia de trabalho consistiu na construção de dois sistemas em escala de laboratório, um leito em canal aberto e um reator de leito empacotado. Estudos foram conduzidos para avaliar a capacidade de neutralização e a qualidade do efluente tratado, considerando os seguintes parâmetros: pH, Fe, Al, Cr, Mn, Zn, Ca, Mg, dureza, sulfatos, condutividade e efeitos toxicológicos para microcrustáceos e peixes. Adicionalmente, foram conduzidos estudos de caracterização da escória antes e após aplicação na neutralização da DAM, visando sua futura destinação. O uso de reator de leito empacotado para tratamento da DAM obteve melhores resultados do que o sistema de canal aberto. O reator evita a percolação do efluente por caminhos preferenciais, aumentando a relação volume tratado/volume de leito. Os ensaios realizados com a escória do forno-panela no reator de leito empacotado demonstraram que esse material apresenta uma alta basicidade, permitindo equalização do efluente de saída para que seja descartado na faixa neutra de pH com a remoção dos metais, enquanto com calcário a remoção dos metais foi parcial, ficando o Fe e Mn com valores acima do permitido pela legislação para o descartes de efluentes. Em ambos os casos, o efluente tratado não apresentou toxicidade. Um dos problemas identificados, para ambos os materiais, foi o recobrimento das partículas pelos hidróxidos metálicos na superfície dos materiais, que reduziu a vida útil dos sistemas e o volume de efluente tratado. O ensaio de expansibilidade demonstra que a escória do forno-panela, na sua forma original, apresenta característica expansiva. Após a neutralização do efluente, a escória apresentou expansão menor que o limite estabelecido na norma ASTM C1260, o que poderia viabilizar sua utilização como agregado na construção civil. Os estudos realizados neste trabalho permitiram concluir que o uso de escórias do forno-panela para neutralização de DAM é uma solução, no sentido de melhorar a qualidade da água em depósitos de rejeitos de carvão, possibilitando um destino para a escória de aciaria elétrica após o tratamento da DAM. / The acid mine drainage (AMD) is formed when certain sulfide minerals are exposed to oxidizing conditions. The AMD is commonly associated with coal mining, especially in coal waste deposits that contain high pyrite (FeS2) concentration. The neutralization of AMD using alkaline materials, like limestone and steel slag is one of the useful remediation techniques. The aim of this work was to study the performance of an electric steel slag of a ladle furnace in the neutralization of the AMD in a percolation bed in a comparative way with limestone, material normaly used for this purpose. The methodology of this work is based in the set up of two systems at labotatory scale, one with open channel bed and the other with bed packed reactor. The studies were directed in order to evaluate the neutralization capacity and the quality of the treated effluent, considering the following parameters: pH, Fe, Al, Cr, Mn, Zn, Ca, Mg, hardness, sulphate, conductivity, and toxicological efects to microcrustacean and fish. Addicionaly, some studies was carried out to caracterize the slag before and after application in the neutralization of the AMD, looking for a future destination. The use of the packed bed reactor for the AMD treatment showed better results than the open channels. The reactor avoids the percolation of the effluent to preferential ways, increasing the volume treated/volume of the bed ratio. The tests carried out with slag of the ladle furnace in the packed bed reactor proves that this material have great basicity allowing the equalization of the effluent output to be discharged in a neutral pH degree with metals removal, while with limestone the metals upstake is partial, remainig Fe and Mn with values above than the allowed by the legislation to effluent discharge. In both cases, the efluent didn’t show toxicity. One of the identified problems to both materials was the metalic hidroxid recovering in the surface, reducing the useful life of the system and the volume of treated effluent. The expansibility tests showed that the ladle furnace slag in the original form have expansive characterists. After the neutralization of the effluent, the slag expansion remains below of the standard border ASTM C1260, and could be possible the use as an aggregate in the civil construction. The results showed in this work allows the conclusion that the use of the ladle furnace slag for neutralization of the AMD is a possible solution to improve the water quality in coal waste deposits.

A mineração de carvão gera grandes volumes de rejeitos que podem ser responsáveis por graves danos ambientais. A oxidação da pirita (FeS2), na presença de ar e água, promove a formação da drenagem ácida de mina (DAM), uma solução aquosa fortemente ácida e rica em sulfato e ferro (nas formas Fe²+ e Fe³+), além de outros metais associados. Atualmente, poucos estudos têm considerado a possibilidade da produção de materiais de valor econômico agregado a partir da água de percolação em rejeitos de carvão. Neste trabalho, o objetivo foi o desenvolvimento de uma rota hidrometalúrgica para a produção de sulfato ferroso (FeSO4). O trabalho experimental consistiu inicialmente na coleta de amostra de rejeitos de carvão rico em pirita e a sua caracterização. Realizou-se, em laboratório, a lixiviação do material em colunas de percolação em ambiente oxidante, adequado para proporcionar a oxidação da pirita em meio aquoso. A recirculação da lixívia permitiu a obtenção de um extrato rico em ferro. A seguir, procedeu-se a conversão do Fe³+ em Fe²+ para obtenção do sulfato ferroso, utilizandose radiação ultravioleta. O lixiviado, rico em Fe+², foi evaporado para cristalização do sulfato ferroso e purificado com álcool etílico. Os cristais foram caracterizados por análise química elementar, difração de raios X e microscopia eletrônica de varredura. A recuperação de Fe na forma melanterita (FeSO4.7H2O) em relação ao Fe pirítico existente na coluna (0,30 kg de Fe pirítico por kg de rejeito de carvão) variou de 7,5 a 9,0 %. Os resultados demonstraram que é possível produzir sulfato ferroso n-hidratado tendo como matéria-prima o rejeito rico em pirita resultante da mineração de carvão mineral. / The coal mining generates large volumes of tailings that may be responsible for serious environmental damages. The oxidation of pyrite (FeS2), in the presence of air and water, promotes the formation of acid mine drainage (AMD), an aqueous and highly acid solution rich in sulfate and iron (in the form Fe³+ and Fe²+), along with other associated metals. Currently, few studies have considered the possibility of production of materials with agregated economic value from the percolation of water in coal mining tailings. In this study, the objective was the development of a hydrometallurgical route for its production. The experimental work consisted initially in the collection of samples of coal waste rich in pyrite and its characterization. In the laboratory was performed the leaching of the material in percolation collums in an oxidizing environment, appropriate to provide the pyrite oxidation in aqueous medium. The recirculation of liquor allowed to obtain an extract rich in the iron. Then, the conversion of Fe³+ to Fe²+ for obtaining ferrous sulfate was performed using ultraviolet irradiation. The leached liquor, rich in Fe+², was evaporated for ferrous sulphate crystallization and purified with ethanol. The ferrous sulphate crystals were characterized by x ray diffraction and scanning electron microscopy. The recovery of Fe in the form melanterite (FeSO4.7H2O ) on pyrite Fe existing in the column (0,30 kg of Fe per kg of pyrite tailings and coal) ranged from 7,5 to 9,0 %. The results demonstrated that it is possible to produce nhydrated iron sulfate having as a raw material the waste rich in pyrite produced from the coal mining.

A mineração de carvão gera milhões de toneladas de rejeitos, contendo pirita (FeS2). A oxidação da pirita, na presença de ar e água, dá origem a um importante problema ambiental comum em regiões de mineração de carvão, a geração de drenagem ácida de mina (DAM). A oxidação da pirita em presença de água promove a formação de H2SO4 (ácido sulfúrico) o que promove a liberação de ferro nas formas de Fe+2 e Fe+3 , sulfatos e uma série de outros metais como Al, Mn e Zn. Esse problema ocorre de forma bastante intensa na região carbonífera do sul do Brasil em especial nos Estados de Santa Catarina e Paraná. A DAM produzida pelos rejeitos da mineração de carvão necessita passar por um processo de tratamento visando diminuir a acidez e a carga de metais despejada no meio, de forma a atender as exigências ambientais. Assim, o objetivo do presente trabalho foi estudar a produção de um coagulante, o sulfato férrico, a partir dos seguintes materiais: (a) lodo férrico obtido por precipitação seletiva da DAM; (b) rejeito obtido pela jigagem do carvão da Camada Barro Branco, Santa Catarina; e (c) concentrado de pirita obtido pelo rebeneficiamento do rejeito de carvão da Mina do Cambuí, Paraná. A metodologia para a produção do coagulante a partir do lodo consistiu na inicialmente na precipitação seletiva do ferro em pH 3,8, lavagem do precipitado, adição de ácido sulfúrico e evaporação para obtenção de uma solução com concentração de Fe3+ de até aproximadamente 12%. A metodologia para a obtenção do coagulante a partir da pirita presente no rejeito de carvão ou no concentrado de pirita consistiu na percolação de água sob condições oxidantes para a produção de uma lixívia rica em sulfato férrico. A lixívia também foi evaporada para obtenção de uma solução de sulfato férrico com concentrações de Fe3+ de até aproximadamente 12%. A qualidade do coagulante produzido foi avaliada em termos da concentração de Fe, Al, Ca, Mn, Zn, Cu e sulfatos. Os resultados demonstram que todos os materiais estudados apresentaram potencial de produção do coagulante. Entre os contaminantes encontrados, destacam-se os elementos alumínio, cálcio, manganês e zinco. Entretanto, quanto maior o teor de enxofre da amostra, maior o rendimento e maior pureza do produto. Concluiu-se que o beneficiamento do rejeito de carvão para a concentração da pirita permite a obtenção de sulfato férrico de melhor qualidade. Os coagulantes produzidos foram utilizados no tratamento do esgoto do Campus do Vale da UFRGS e de água de abastecimento da região de Porto Alegre. As idéias de valorização dos rejeitos da mineração de carvão estão em sintonia com os princípios da atualidade no que diz respeito ao desenvolvimento sustentável e aos programas de valoração de resíduos e produção mais limpa. / The coal mining generates millions of tonnes of tailings containing pyrite (FeS2). The oxidation of pyrite in the presence of air and water, gives rise to an important environmental problem common in areas of coal mining, the generation of acid mine drainage (AMD). The oxidation of pyrite in the presence of water promotes the formation of H2SO4 (sulfuric acid) which promotes the release of iron in the form of Fe +2 and Fe +3, sulphates and a host of other metals such as Al, Mn and Zn. This problem occurs quite intense in the coal in southern Brazil in particular in the states of Santa Catarina and Parana. The DAM produced by coal waste mining need to go through a treatment process in order to decrease the acidity and metal load discharged into the environment, in order to meet environmental requirements. The objective of this work was to study the production of a coagulant, ferric sulfate, with the following materials: (a) ferric sludge obtained by selective precipitation of DAM (b) obtained by reject coal concentration of the Layer Barro Branco, Santa Catarina, and (c) pyrite concentrate obtained by concentration of the tailings coal mine of Cambuí, Paraná. The methodology for the production of coagulant from sludge was the first in the selective precipitation of iron in pH 3.8, washing the precipitate with sulfuric acid and evaporation to obtain a solution with Fe 3 + concentration of approximately 12% with Fe2O3. The methodology for obtaining the coagulant from the pyrite present in coal or reject the pyrite concentrate was the percolation of water under oxidizing conditions to produce a liquor rich in ferric sulphate. Leach was also evaporated to obtain a solution of ferric sulphate with concentrations of Fe3 + up to approximately 12%. The quality of the coagulant was evaluated in terms of concentration of Fe, Al, Ca, Mn, Zn, Cu and sulfates. The results show that all the materials studied showed potential for production of the coagulant. Among the contaminants found, highlight the elements aluminum, calcium, manganese and zinc. However, the higher the sulfur content of the sample, the higher the yield and higher purity of the product. It was concluded that the processing of the waste coal to the concentration of pyrite to obtain ferric sulphate better. Coagulants produced were used in the treatment of sewage Campus do Vale UFRGS and water supply in the region of Porto Alegre. Ideas for use of tailings from coal mining are in line with the principles of the present time regarding the development and evaluation programs for waste and cleaner production.

Este trabalho teve como objetivo avaliar o desempenho de diferentes tipos de coberturas secas utilizadas em células experimentais preenchidas com rejeitos piritosos de carvão mineral. Quatro células de igual dimensão foram preenchidas com rejeitos piritosos, provenientes do beneficiamento de carvão mineral da Unidade Mineira II – Verdinho, pertencente à Carbonífera Criciúma S.A., situada no município de Forquilhinha, Estado de Santa Catarina. Em relação à composição das células, a primeira foi construída sem cobertura; a segunda, com somente pré-cobertura, constituída de mistura de rejeitos; a terceira, com cobertura de argila compactada; e a quarta elaborada com sistema de barreira capilar dupla. Todas foram expostas às mesmas condições meteorológicas. Para avaliação do desempenho das coberturas analisou-se o volume e características químicas e físico-químicas da água de percolação no período compreendido entre 2009 e 2010. Os parâmetros físicoquímicos considerados foram os seguintes: pH, Eh, acidez, alcalinidade e condutividade enquanto que os parâmetros químicos foram Fe, Al, Mn, Zn e sulfato. A partir dos resultados obtidos, fez-se uma análise técnica dos diferentes tipos de cobertura seca utilizados. De acordo com análise técnica constatou-se que o melhor desempenho no tocante ao volume e à qualidade da água percolada ocorreu na célula em que se utilizou cobertura com argila compactada e na célula em que se empregou cobertura do tipo barreira capilar dupla. / This work aimed to evaluate the performance of different types of dry cover employed in experimental cells filled with pyritic waste of mineral coal. Four cells with the same dimension were filled with coal tailings from the coal preparation plant of Unidade Mineira II –Verdinho, which belongs to Carbonífera Criciúma SA, located in Forquilhinha, Santa Catarina State. The first cell was constructed without any kind of cover, the second one was built with a cover composed by a mixture of coal wastes, the third one was built with a layer of compacted clay, and the forth one was constructed with a system composed by a dual-capillary barrier. All were exposed to the same meteorological conditions. To evaluate the performances of dry covers we analyzed the volume and the chemical and physicochemical characteristics of seepage between 2009 and 2010. The water was analyzed in terms of pH, Eh, acidity, alkalinity, conductivity and the concentration of Fe, Al, Mn, Zn, and sulfate. The experimental results showed that the best performance concerning the volume and the quality of the percolated water was obtained in the cell that received a cover of compacted clay and in the one that received a cover composed by dualcapillary barrier.

The environmental impact of the Robinson Deep mine residue deposit in Johannesburg was assessed by evaluating the water chemistry and surface precipitates, thereby identifying the processes responsible for forming the contamination footprint. Precipitates were compared to a documented paragenetic sequence of mineral formation associated with sulphide rich mine waste. PHREEQC, a geochemical modelling tool, was utilized to predict the formation of precipitates from evaporation ponds. The chemistry of the leachate was analysed and compared to water quality standards in order to determine the possible environmental impact. The chemistry of the water emanating from the mine residue deposit reveals that an Fe-SO4 dominant chemistry persists, this is consistent with acid mine drainage environments. The most toxic cations and anions contained in the water are Fe, Al, Mn, Ca, As, Mg, Cu, Zn, Pb and SO . It is predicted that the impact of toxic metals identified in the water decreases further from the mine residue deposit due to dilution and co-precipitation with different mineral phases such as goethite. Precipitates identified include jarosite group minerals, goethite, melanterite, copiapite, Mg-copiapite, halotrichite, pickeringite, gypsum and alunogen. These secondary minerals may be used as indicator minerals of acid mine drainage. Assessment and prediction of the stage of contamination and possible environmental impact, may, therefore be pursued when comparing the indicator minerals to predicted paragenetic sequences. For example, the precipitation of melanterite is consistent with an early stage of acid mine drainage development. The presence of melanterite thus suggests that oxidation of sulphides is an ongoing process on the Robinson Deep mine residue deposit. Evaporation of water in the evaporation ponds aids in increasing the concentration, hence allowing the predominant precipitation of jarosite group minerals. Alternatively, rainfall dilutes the water allowing dissolution of minerals located on the banks of the evaporation ponds to predominate. This mechanism of precipitation and dissolution is seasonal; formation of precipitates predominates during the dry season, while dissolution is most prominent during the wet season. The development of hardpans indicates that the main mechanism of formation of the associated phases is through capillary action at the sediment surface. Leaching of Fe2+-rich water from the mine residue deposit containment area is indicated by the presence of copiapite, while jarosite and goethite formation tend to form part of a hardpan layer. Goethite is an indication of a late stage mineral predominant at lower sulphate and higher pH conditions. Results of predicted formation of precipitates by PHREEQC are not in very good agreement with actual field observations. This is mainly due to the lack of thermodynamic data for many of the sulphate minerals observed. Hence, precipitates associated with acid mine drainage may be utilized as indicator minerals. Consequently, there identification may facilitate in environmental monitoring and risk assessment. / Prof. J. M. Huizenga Prof. J. Gutzmer

Les liens entre la diversité et l'adaptation des populations bactériennes à leur environnement constituent une problématique importante en écologie microbienne. L'accès à de nombreux génomes permet aujourd'hui des avancées intéressantes dans ce domaine. Plusieurs souches du genre Thiomonas et appartenant à la même espèce, Tm. arsenitoxydans 3As et Tm. spp. CB1, CB2, CB3 et CB6, ont été isolées d'un drainage minier acide (DMA) à Carnoulès (Gard). La comparaison de leur génome a permis d'affiner leur phylogénie et de mettre au jour des différences de contenu génétique liées à des îlots génomiques. Certaines de ces différences ont été corrélées expérimentalement avec des différences fonctionnelles concernant l'oxydation de l'arsénite (As(III)), la dégradation de l'urée et la biosynthèse de biofilm, et confèrent potentiellement un avantage sur le site (meilleure résistance à l'As(III), précipitation des métaux et augmentation du pH, protection des cellules). La comparaison de la synténie des génomes de Tm. arsenitoxydans 3As avec Tm. sp CB2 et Tm. intermedia K12 (non isolée de ce DMA) a montré que le génome de Tm. sp. CB2 a subi plusieurs remaniements importants. Ces types de réarrangements pourraient être en partie à l'origine de l'apparition de variants "super-résistants" à l'As(III) dans la population. En particulier, plusieurs copies d'un élément intégratif et conjugatif portant l'opéron aioBA codant l'arsénite oxydase ont été détectées chez deux variants, ce qui pourrait en partie expliquer leur résistance accrue à l'As(III). La proportion de variants est plus importante en présence d'As(III) au sein des biofilms, et des données de transcriptomique ont en effet montré que ces remaniements seraient en partie causés par des systèmes de réparation de l'ADN suite à des dommages liés au stress oxydant induit par l'As(III). Le développement en biofilm et la présence d'As(III) modulerait donc la flexibilité génomique et le potentiel adaptatif de Tm. sp. CB2. Ces données suggèrent que cette souche, avec Tm. sp. CB3, possèdent un génome visiblement plus flexible que les autres. Les souches Tm. arsenitoxydans 3As et Tm. spp. CB1 et CB6, contrairement à Tm. spp. CB2 et CB3, forment un groupe distinct d'un point de vue phylogénétique et fonctionnel ("groupe 3As"), suggérant qu'elles occupent une niche écologique spécifique et pourraient constituer un écotype. La population de Thiomonas de ce DMA serait donc composée d'au moins un écotype stable et de souches au génome plus instable, dont le potentiel adaptatif plus important serait influencé par l'As(III) et le développement en biofilm. / Understanding the link between diversity and adaptation in natural bacterial populations represents an important issue in microbial ecology. The amount of whole genome sequencing data currently available has allowed for interesting advances in this field. Several strains from the genus Thiomonas belonging to the species, Tm. arsenitoxydans (3As) and Tm. spp. (CB1, CB2, CB3 and CB6), were isolated from the acid mine drainage (AMD) at Carnoulès (Gard, France). Comparison among genomes allowed for a better definition of their phylogenetic relationships and highlighted differences in genetic content, which is essentially due to the presence of genomic islands. Some of these differences were experimentally correlated with functional traits concerning arsenite oxidation, urea degradation, and biofilm biosynthesis, and are potentially beneficial in situ (leading to an enhanced resistance to arsenite (As(III)), metal precipitation, and an increase in pH, ultimately protecting cells). The comparison of genome synteny of Tm. arsenitoxydans 3As, Tm. sp CB2, and Tm. intermedia K12 (not isolated from this AMD) show several important genomic rearrangements exist in Tm. sp CB2. This type of rearrangements could be involved in the emergence of arsenite "super-resistant" variants in the Tm. sp CB2 population. In particular, several copies of an integrative and conjugative element (ICE) containing the aioBA operon coding arsenite oxidase were detected in the genomes of two variants, which could explain their higher levels of resistance to As(III). The percentage of variants in biofilm culture is higher when grown in the presence of As(III), and transcriptomic data suggests that genomic rearrangements probably occurred through DNA repair systems following damage caused by As(III) induced oxidative stress. Therefore, biofilm development and As(III) appear to allow for the adaptation of Tm. sp. CB2 genome flexibility and evolutionary potential. These data suggest that CB2 and Tm. sp. CB3 have more flexible genomes than the other strains. Tm. arsenitoxydans 3As and Tm. spp. CB1 and CB6 form both a phylogenetic and functional cluster ("3As group") suggesting that they occupy a specific ecological niche and therefore could represent an ecotype of the genus Thiomonas. The Thiomonas population from the Carnoulès AMD might therefore consist of at least one stable ecotype as well as other strains with more instable genomes, whose higher adaptive potential could be affected by As(III) and biofilm development.

Acid mine drainage is a well known environmental pollutant, not only in South Africa, but throughout the world, and the use of microbial processes in the treatment of these wastes has been the subject of investigation over past decades. Lignocellulose packed-bed reactors have been used in passive treatment systems, and, although effective initially, they show early decline in performance while the packing material remains largely un-utilized. Little is known about this phenomenon which remains a severe constraint in the development of efficient passive mine water treatment systems. It has been proposed that the degradation pathways of the complex lignocellulose substrate may be limited in some way in these systems during the manifestation of this effect. This study has addressed the problem using a molecular microbial ecology methodology in an attempt to relate trophic functions of the microbial population to the physico-chemical data of the system. A field-scale lignocellulose packed-bed reactor located at Vryheid Coronation Colliery (Northern Kwa-Zulu Natal province, South Africa) was monitored for six years and the results showed the classic profile of performance decline related to a slowdown in sulphate reduction and alkalinity production. The reactor was decommissioned , comprehensive samples were collected along the depth profile and the microbial populations investigated by means of 16S rRNA gene methodology. The population was found to include cellulolytic Clostridia spp., CytophagaIFlavobacterlBacteroidetes, Sphingomonadaceae and as yet uncultured microorganisms related to microbiota identified in the rumen and termite gut. These are all known to be involved as primary fermenters of cellulose. Oesulphosporosinus was present as sulphate reducer. A comparison of substrata sampling and population distribution suggested that spatial and temporal gradients within the system may become established over the course of its operation. Based on these findings, a laboratory-scale reactor was constructed to simulate the performance of the packed-bed reactor under controlled experimental conditions. The laboratory-scale reactor was operated for 273 days and showed comparable performance to that in the field in both biomolecular and physicochemical data. Clearly defined trophic niches were observed. These results suggested that a sequence of events does occur in lignocellulose degradation over time. Based on the spatial and temporal column studies, a descriptive model was proposed to account for these events. It was found that fermentative organisms predominate in the inlet zone of the system using easily extractable compounds from the wood, thus providing feedstock for sulphate reduction occurring in the succeeding compartments. Production of sulphide and alkalinity appears to be involved in the enhancement of lignin degradation and this, in turn, appears to enhance access to the cellulose fraction. However, once the readily extractables are exhausted, the decline in sulphide and alkalinity production leads inexorably to a decline in the overall performance of the system as a sulphate reducing unit operation. These observations led to the proposal that with the addition of a limited amount of a readily available carbon source, such as molasses, in the initial zone of the the reactor, the ongoing generation of sulphide would be sustained and this in turn would sustain the microbial attack on the lignocellulose complex. This proposal was tested in scale-up studies and positive results indicate that the descriptive model may, to some extent, provide an account of events occurring in these systems. The work on sustaining lignocellulose degradation through the maintenance of sulphate reduction in the initial stages of the reactor flow path has led to the development of the Degrading Packed-bed Reactor concept and that, has subsequently been successfully evaluated in the field.

Magister Scientiae - MSc / This study evaluated SO4 2- removal from circumneutral mine water (CMW) collected from Middleburg coal mine using coal FA collected from Hendrina power station. The following parameters were investigated: the effect of the amount of FA, the effect of the final pH achieved during treatment, the effect of the initial pH of the mine water and the effect of Fe and Al on SO4 2- removal from mine water. The precipitation of ettringite at alkaline pH was evaluated to further reduce the SO4 2- concentration to below the DWAF limit for potable water. Removal of SO4 2- from mine water was found to be dependent on: the final pH achieved during treatment, the amount of FA used to treat the mine water and the presence of Fe and Al in the mine water. Treatment of CMW using different CMW:FA ratios; 5:1, 4:1, 3:1, and 2:1 resulted in 55, 60, 70 and 71 % SO4 2- removal respectively. Treatment of CMW to pH 8.98, 9.88, 10.21, 10.96, 11.77 and 12.35 resulted in 6, 19, 37, 45, 63 and 71 % SO4 2- removal respectively. When the CMW was modified by adding Fe and Al by mixing with Navigation coal mine AMD and treated to pH 10, 93 % SO4 2- removal was observed. Further studies were done to evaluate the effects of Fe and Al separately. Treatment of simulated Fe containing AMD (Fe-AMD) to pH 9.54, 10.2, 11.8, and 12.1 resulted in 47, 52, 65, and 68 % SO4 2- removal respectively. When Al containing AMD was treated to pH 9.46, 10.3, 11.5 and 12 percentage SO4 2- removal of 39, 51, 55 and 67 % was observed respectively. / South Africa