Ehemalige Urangruben tragen durch das anfallende Flutungswasser maßgeblich zur Ausbreitung von Schadstoffen wie Uran und Arsen in teils dicht besiedelte Gebiete bei. Um die Prozesse in den unterirdischen Gruben besser zu verstehen und alternative Strategien zur konventionellen, kostenintensiven Wasserbehandlung entwickeln zu können, war das Ziel der Arbeit, mikrobielle Gemeinschaften aus drei gefluteten Uranerzbergwerken in Sachsen, namens Pöhla, Schlema und Zobes, die unterschiedliche Flutungsstadien repräsentierten, zu charakterisieren und den mikrobiellen Einfluss auf die Mobilität von Uran und Arsen zu untersuchen. Um herauszufinden, welche Mikroorganismen die hydrochemischen Vorgänge im Untergrund der Uranerzbergwerke beeinflussen könnten, wurde die Diversität und Zusammensetzung mikrobieller Gemeinschaften mittels Pyrosequenzierung eines Fragments des 16S rRNA Gens (16S rDNA) und CARD-FISH ermittelt. Wenngleich Clusteranalysen zeigten, dass sich die planktonischen Gemeinschaften hinsichtlich ihrer bakteriellen Zusammensetzung zwischen den drei Uranerzbergwerken unterschieden, wurden alle von chemolithotrophen Schwefeloxidierern der Beta- und Epsilonproteobacteria dominiert, die mit den Gattungen Thiobacillus und Sulfuritalea bzw. Sulfuricurvum und Sulfurimonas vertreten waren. Im Unterschied zu den planktonischen Gemeinschaften bestanden in situ Biofilme, die auf BACTRAPs während einer 3-monatigen Exposition im Flutungswasser anwuchsen, laut Pyrosequenzierung zu einem wesentlichen, mitunter dominanten Anteil aus metall- bzw. sulfatreduzierenden Deltaproteobacteria. In Biofilmgemeinschaften aus Zobes wurden hauptsächlich Geobacter sp. detektiert, die als Fe(III)- und U(VI)-Reduzierer bekannt sind. Obwohl Archaea basierend auf den Ergebnissen der CARD-FISH-Analyse nur einen sehr geringen Anteil der planktonischen Gemeinschaften ausmachten, wurden mittels Pyrosequenzierung planktonische Euryarchaeota der Thermoprotei in allen Gruben detektiert. In planktonischen Gemeinschaften und 3-monatigen Biofilmen aus Pöhla und Zobes wurden zudem methanogene Crenarchaeota, vor allem Methanobacteria und teilweise Methanomicrobia, ermittelt. Die 16S rRNA-Analyse, die ergänzend zum DNA-basierten Ansatz durchgeführt wurde, lieferte Hinweise darauf, dass die detektierten, dominanten Mikroorganismen, Bacteria sowie Archaea, in der planktonischen Gemeinschaft aus Schlema und den Biofilmgemeinschaften stoffwechselaktiv waren. In der planktonischen Gemeinschaft aus Zobes wurden im Vergleich zur DNA-basierten Analyse höhere Abundanzen für Verrucomicrobia, Acidobacteria und Alphaproteobacteria ermittelt, deren Bedeutung offen bleibt. Untersuchungen zum mikrobiellen Stoffwechselpotential planktonischer Gemeinschaften mittels CFU- und MPN-Analysen ergaben, dass Mikroorganismen aus allen Urangruben ein breites Spektrum anaerober Reaktionen (Nitrat-, Eisen-, Mangan-, Arsenat- und Sulfatreduktion und Acetogenese) unter Laborbedingungen abdeckten. In guter Übereinstimmung mit den Sequenzierungsergebnissen konnten methanogene Mikroorganismen nur im Flutungswasser aus Pöhla und Zobes detektiert werden. Die Metaproteomanalyse ergab, dass 61,6% der Peptide in der planktonischen Gemeinschaft aus Schlema von den dominanten Epsilonproteobacteria stammten. Dagegen wurden für Zobes detektierte Peptide mehrheitlich methylotrophen und eisenoxidierenden Betaproteobacteria der Familien Methylophilaceae bzw. Gallionellaceae sowie methylotrophen Gammaproteobacteria der Methylococcaceae zugewiesen. Obwohl die Mehrheit der Proteine an der Translation beteiligt war, konnten insgesamt 49 Proteingruppen ermittelt werden, deren Vertreter für den mikrobiellen Energiestoffwechsel relevant waren. Insbesondere planktonische Gammaproteobacteria aus Zobes konnten so mit dem Kohlenstoff- und Schwefelkreislauf in Zusammenhang gebracht werden. Mithilfe von Labormikrokosmen wurde der potentielle Einfluss mikrobieller Gemeinschaften aus Schlema auf die Mobilität von Arsen und Uran im Flutungswasser mit Acetat als Elektronendonor unter anaeroben Bedingungen über einen Zeitraum von 98 Tagen untersucht. Im Vergleich zu den Kontrollen konnten sowohl die stimulierte, planktonische Gemeinschaft als auch Biofilme natürliches Arsen aus der wässrigen Phase fast vollständig entfernen. Allerdings wies der spätere Anstieg des gelösten Arsens daraufhin, dass der immobilisierte Zustand langfristig nicht stabil blieb. In stimulierten Biofilm-Ansätzen wurde Uran mit bis zu 39 ± 9% (in Anwesenheit von 7 µM natürlichem Uran) bzw. 34 ± 8% (bei Zugabe von 50 µM U(VI)) aus der wässrigen Phase langfristig (98 Tage) immobilisiert. Laserfluoreszenzspektroskopische Untersuchungen zeigten, dass Uran im Biofilm reduziert wurde.

Microbial diversity plays an important role in the decomposition of soil organic matter. However, the drivers of this dependence still remain unclear. The work is based on long-term monitoring of soils of different successional stages of different diversity. Soil sampling was conducted on two dumps after brown coal mining in the Sokolov. Soils were X ray sterilized and inoculated by inoculum from both soils in two inoculation density which create gradient of microbial diversity. Then microbial respiration was measured in either supplied or not supplied by litter of Calamagrostis epigejos. Results showed a strong positive correlation between the microbial diversity and decomposition of organic matter if the microbial community is limited by available carbon. If there isn't carbon limit available, the decomposition rate is controlled by the amount of microbial biomass. Results demonstrated positive correlation between the rate of decomposition and the amount of fungal biomass. The soils with the addition of leaf litter showed priming effect in the initial stage of decomposition. In the control samples without addition of litter priming effect wasn't observed. Increasing humidity led to increase of decomposition rate. We can conclude with a clear conscience that similarly conclusive results associated...

O Rio Tietê e o rio mais importante do Estado de São Paulo e um dois mais poluídos do Brasil. No seu percurso ele recebe altas concentrações de poluentes orgânicos e industriais que podem alterar as comunidades microbianas autóctones. Uma análise dos perfis metagenômicos realizada em pontos do Rio Tietê com diferentes qualidades de água, permitiu observar a influencia dos parâmetros ambientais na composição e funcionamento das comunidades microbianas presentes no rio. Locais eutróficos mostraram prevalência de micro-organismos e funções associadas à heterotrofia. Em contrapartida, nos locais oligotróficos foram detectadas comunidades microbianas e funções relacionadas a metabolismos autótrofos e litótrofos. Foi visto também que as pressões ambientais podem promover a aquisição, por parte dos micro-organimos, de características que lhes permitam sobreviver às condições ambientais adversas, como a tolerância a metais tóxicos. / The Tietê River is the most important river of the State of São Paulo, in the southeastern of Brazil; it is also one of the most contaminated rivers of the country. On its watercourse, the river receives large amounts of anthropogenic and industrial pollutants that can alter the environmental conditions of the water, leading to possible shifts in the microbial diversity. Metagenomic profiles of river locations with different water qualities showed taxonomic and functional differences related to the sampling site and water quality. Eutrophic sites showed prevalence of microorganisms and functions associated with heterotrophy, in contrast, in oligotrophic sites were detected microbial communities and functions related to autotrophic and litotrophic metabolisms. Also was observed, that environmental pressures can promote the acquisition of characteristics that allow the local microorganism to survive to adverse environmental conditions, such as tolerance to toxic metals.

Os impactos da poluição por hidrocarboneto de petróleo sobre a diversidade e funcionalidade das comunidades microbianas em manguezais não são totalmente conhecidos, principalmente devido às limitações metodológicas para acessar os microrganismos nãocultiváveis. No entanto, vários métodos moleculares independentes de cultivo têm sido utilizados para investigar a diversidade e a estrutura das comunidades microbianas em ecossistemas naturais. O objetivo deste trabalho foi avaliar as variações da estrutura das comunidades de Bacteria e Archaea e a diversidade de Bacteria em uma transeção de solo de mangue do rio Iriri (Bertioga, SP) com um gradiente de contaminação por hidrocarbonetos de petróleo. As análises por eletroforese em gel com gradiente desnaturante (DGGE) mostraram que as comunidades de Bacteria e Archaea nas diferentes posições geográficas foram mais similares entre si do que entre diferentes profundidades ao longo do perfil em uma mesma posição geográfica. A análise das seqüências de clones de rDNA 16S de Bacteria dos diferentes pontos amostrados em abril de 2000, mostrou que a diversidade genética, avaliada pelo índice de Shannon, das comunidades microbianas diferem estatisticamente somente entre ponto o P1 (ponto menos contaminado) e P3 (ponto mais contaminado). As estimativas não-paramétricas da riqueza de espécies mostraram que P1, P2 e P3 possuem mais de 3539, 2524 e 1421 espécies bacterianas, respectivamente. Já, para as amostras do ponto P2 coletadas nos anos 2000 e 2004, muito embora os valores dos índices de Shannon tenham sido semelhantes, houve uma provável dominância de grupos específicos nas amostras coletadas em 2004, verificada pelos altos valores da recíproca do índice de Simpson. Os dados mostraram também que o número estimado de espécies bacterianas no ponto P2 diminuiu com o tempo, sendo menor em amostras de 2004, se comparado com amostras de 2000. No geral, a afiliação filogenética dos clones de rDNA 16S mostrou a grande diversidade de espécies, a maioria não conhecidas. Os dados sugerem que a contaminação do solo de mangue do rio Iriri está selecionando microrganismos mais adaptados às fontes de carbono introduzidas no solo. / The impacts of petroleum hydrocarbon pollution on the diversity and functionality of the microbial communities in mangrove soils are not totally understood, mainly due to the methodological limitations to access unculturable microorganisms. However, several cultureindependent molecular methods have been used to investigate the diversity and structure of microbial communities in natural ecosystems. The aim of this work was to evaluate shifts in Bacteria and Archaea community structures and the diversity of Bacteria in a soil transection of the Iriri river mangrove (Bertioga, SP) showing a petroleum hydrocarbon contamination gradient. The analyses by denaturing gradient gel electrophoresis (DGGE) showed that the communities of Bacteria and Archaea in different geographical positions were more similar among them than the communities in different depths along the soil profile at the same geographical position. Sequence analyses of bacterial 16S rDNA clones from different points sampled in April 2000 showed that the genetic diversity of the bacterial communities, based on the Shannon index, differ statistically only between P1 (less polluted) and P3 (more polluted) locations. Nonparametric estimates of species richness showed that P1, P2 and P3 may have more than 3539, 2524 and 1421 bacterial species, respectively. For P2 sampled in years 2000 and 2004, even though the Shannon indices were similar, there was a probable dominance of specific bacterial groups in year 2004, based on the high values of the reciprocal of Simpson\'s index. The data also showed that the estimated number of bacterial species in P2 decreased with the time, being lower in samples collected in 2004, as compared to samples collected in 2000. In the general, the phylogenetic affiliation of the 16S rDNA clones showed high bacterial species diversity, and most of the bacteria were of unknown species. The data suggest that the contamination of Iriri river mangrove soil with petroleum hydrocarbon is selecting microorganisms more adapted to the introduced carbon sources into the soil.

Vulcões ativos na Antártica contrastam com a paisagem predominantemente gelada do continente. Eles fornecem condições únicas capazes de selecionar uma grande variedade de adaptações microbianas. A Ilha Deception localiza-se na região da Península Antártica e difere de outros vulcões antárticos especialmente pela influência marinha e temperaturas mais elevadas. Foram coletadas amostras de sedimentos associados a fumarolas e geleiras em dois sítios geotermais de Deception, com temperaturas variando entre 0°C a 98°C. Diferentes técnicas independentes de cultivo foram empregadas com o intuito de entender como as comunidades microbianas respondem as variações ambientais extremas produzidas pela atividade vulcânica. Os resultados indicaram que a co-ocorrência de arqueias hipertermófilas e suas adaptações com micro-organismos metabolicamente diversos adaptados a regiões geladas representa uma estrutura de comunidades única para ecossistemas antárticos. Este trabalho forneceu dados ineditos sobre questoes centrais de diversidade e adaptacao microbiana a ambientes geotermais polares. / Active volcanoes in Antarctica contrast with the predominately icy landscape. They harbor unique conditions capable to select an extreme range of microbial adaptations. Deception Island is located in the Antarctic Peninsula region and differs from other Antarctic volcanoes specially by its higher temperatures and marine influence. We collected sediment samples associated to active fumaroles and glaciers on two geothermal sites of Deception Island, with temperatures ranging from 0°C to 98°C. Different cultivation-indepedent techniques were used to understand how microbial communities respond to extreme environmental variations produced by volcanic activity. The results indicate that co-occurrence of hyperthermophiles and their specific adaptations with metabolically diverse cold-adapted micro-organisms represents a unique community structure for antarctic ecosystems. This study provided primordial data on central questions about microbial diversity and adaptation to polar geothermal environments.

Made available in DSpace on 2017-07-21T18:53:09Z (GMT). No. of bitstreams: 1 Maria Isabel Stets.pdf: 1158333 bytes, checksum: c7c568acfaf2daf456026f86016faf6b (MD5) Previous issue date: 2008-03-12 / The effluent generated in abattoirs has a high concentration of organic matter and need treatment before being discarded in the water bodies. Anaerobic biological filters are considered a good technique for the treatment of industrial effluents; however, choosing the correct support media is extremely important to ensure the success of the reactor. In the same way, the study of microbial diversity can optimize the performance and the operation of these systems. In this work it was evaluated the efficiency of the treatment of the effluent from slaughterhouse, using three upflow anaerobic biological filters, and it was compared the microbial diversity of these filters. The reactor called A, B and C were built in chloride polivinile and filled with rings of polypropylene, polyurethane foam, or pieces of clay brick, respectively. These reactors were operated at room temperature and the used Hydraulic Retention Times of (HRTs) were of 30, 20, 18, 14, 10, 8, 6, 5, 4, 3, 2, 1.5 and 1 day. To monitor the efficiency of the process, the pH, alkalinity, acidity, Chemistry Oxygen Demand (COD), and total solids content of volatile solids, nitrogen and phosphorus substrate and effluents of each reactor were analyzed. The values of pH decreased with the reduction of HRT and the ratio volatile acidity / alkalinity remained below 0.13 during the majority of the analyzed period. In the one day’s HRT, the reactor C was more effective reaching 80.76 of removal. There was a high rate of nitrogen removal probably due to its low concentration in the substrate. High phosphorus removals also occurred: there was a decrease of phosphorus removal when the HRT declines in reactors A and B, the same did not occur in the reactor C. The three reactors behaved similarly regarding to the collecting sites and there was no difference between these sites when COD removal, volatile acidity / alkalinity ratio and pH were analyzed. To evaluate the effect of different media support in the microbial community, the reactors DNA biomass was extracted, quantified and used as template in an amplification reaction of 16S rDNA gene from bacteria and metanogenic. The amplified fragments of approximately 1500 bp were treated with HinfI, RsaI and HaeIII restriction enzymes to obtain the reactors ARDRA (Amplified Ribossomal DNA Restriction Analysis) profiles. The indices of richness, richness modified, Shannon-Weaver diversity index and similarity dendogram, determined from ARDRA, grouped the reactors A and B, and left the reactor C isolated due to its dissimilarity. The Hierarchical Cluster Analysis (HCA) also grouped the reactors that way, however, that group has less than 0.1% of similarity, suggesting a low interference in the reactors media support in microbial communities. / O efluente gerado em abatedouros possui uma elevada concentração de matéria orgânica e necessita de tratamento antes de ser descartado nos corpos hídricos. Os filtros biológicos anaeróbios são considerados uma boa técnica para tratamento de efluentes industriais, porém, a escolha correta do meio suporte é extremamente importante para garantir o sucesso do reator. Da mesma forma, o estudo da diversidade microbiana pode otimizar o desempenho e a operação desses sistemas. Neste trabalho, foi realizada a avaliação da eficiência do tratamento de efluente de abatedouro, utilizando-se três filtros biológicos anaeróbios de fluxo ascendente, bem como a comparação da diversidade microbiana desses filtros. Os reatores denominados A, B e C foram construídos em cloreto de polivinila e preenchidos com anéis de polipropileno, espuma de poliuretano ou pedaços de tijolo de argila, respectivamente. Esses reatores foram operados à temperatura ambiente e os Tempos de Retenção Hidráulica (TRHs) utilizados foram de 30; 20; 18; 14; 10; 8; 6; 5; 4; 3; 2; 1,5 e 1 dia. Para monitorar a eficiência do processo, foram analisados no substrato e efluente de cada reator pH, alcalinidade, acidez, Demanda Química de Oxigênio (DQO), teor de sólidos totais e sólidos voláteis, nitrogênio efósforo. Os valores de pH do efluente dos reatores decresceram com a redução do TRH. A relação acidez volátil/alcalinidade ficou abaixo de 0,13 durante a maior parte do período analisado. No TRH de 1 dia o reator C foi o mais eficiente alcançando 80,76 de remoção de DQO. Observou-se uma elevada taxa de remoção de nitrogênio em todos os reatores provavelmente devido à sua baixa quantidade no substrato. Também ocorreram elevadas remoções de fósforo, mas houve decréscimo dessa remoção com a diminuição do TRH nos reatores A e B, o mesmo não ocorrendo no reator C. Quando se analisaram as amostras retiradas em quatro diferentes alturas dos reatores, observou-se que os três se comportaram de forma similar quanto aos pontos de coleta e não houve diferença entre esses pontos quanto à remoção de DQO, relação acidez volátil/alcalinidade e pH. Para avaliar o efeito dos diferentes meios suporte na comunidade microbiana, o DNA da biomassa dos reatores foi extraído, quantificado e usado como molde numa reação de amplificação do gene 16S rDNA de bactérias e archaeas metanogênicas. Os fragmentos de aproximadamente 1500 pb amplificados foram tratados com as enzimas de restrição HinfI, RsaI e HaeIII para obtenção dos perfis de ARDRA (Análise da Restrição de rDNA Amplificado) dos reatores. Os índices de riqueza, riqueza modificada, diversidade de Shannon-Weaver e dendograma de similaridade, determinados a partir da ARDRA, agruparam os reatores A e B, ficando o reator C isolado devido à sua dessemelhança. A Análise de Agrupamento Hierárquico (HCA) também agrupou os reatores dessa forma, entretanto, esse agrupamento possui menos que 0,1% de similaridade, sugerindo uma baixa interferência do meio suporte nas comunidades microbianas.

Le présent travail s’intéresse à l’étude de la diversité des communautés procaryotiques, basée sur le gène codant pour l’ARNr 16S et sur les gènes codants pour la sous-unité β du sulfite réductase dissimilatrice (dsrB) et la sous-unité alpha de la méthyl-coenzyme M réductase (mcrA), pour étudier la diversité de la communauté des bactéries sulfato-réductrices et des méthanogènes, respectivement à partir des échantillons collectés en saison sèche ou pluvieuse du Chott El Jerid. Les analyses des séquences du gène codant pour l’ARNr 16S ont montré que les bactéries regroupées aux Proteobacteria et Firmicutes sont détectés dans les deux saisons alors que les séquences appartenant au groupe taxonomique Bacteroidetes, Actinobacteria et Betaproteobacteria sont apparues uniquement dans la saison pluvieuse. Le groupe Deinococcus-Thermus sont observés que dans la saison sèche. Dans le domaine des archées, la plupart des séquences appartiennent au phylum Euryachaeota, détecté dans les deux saisons, alors que, le phylum Crenarchaeota apparait uniquement dans la saison pluvieuse. En plus, les bactéries sulfato-réductrices, appartenant à la classe Deltaproteobacteria, sont fréquents notamment à la saison pluvieuse prouvée déjà par les deux techniques DGGE et qPCR. A partir des cultures d’enrichissement, de nombreuses bactéries anaérobies fermentaires appartiennent aux familles Halanaerobiaceae et Halobacteroidaceae. Les analyses phylogénétiques ainsi que les caractéristiques phénotypiques et physiologiques montrent une nouvelle souche Sporohalobacter salinus proche de l’espèce Sporohalobacter lortetii, seule espèce décrite à ce jour du genre Sporohalobacter. / The present work concerns microbial biodiversity of prokaryotic communities, sulfate-reducing bacteria, and methanogens targeting the 16S rRNA gene and functional gene markers encoding the dissimilatory sulfite reductase β-subunit gene (dsrB) and alpha subunit of the methyl-coenzyme M reductase (mcrA), respectively from samples collected in the dry and wet seasons from Chott El Jerid. Phylogenetic analysis targeting the 16S rRNA gene showed that bacteria were grouped to Proteobacteria and Firmicutes detected at both seasons, whereas, Bacteroidetes, Actinobacteria and Betaproteobacteria were present only in the wet season. Deinococcus-Thermus group were observed in the dry season. Archaeal sequences were belonged to the phyla of Euryarchaeota in both seasons and Crenarchaeota was appeared in wet season. Sulfate-reducing bacteria, related to Deltaproteobacteria class were dominant mainly in wet season proved by two techniques DGGE and QPCR. From enrichment cultures, anaerobic fermentative bacteria were isolated in pure cultures, related to Halanaerobiaceae and Halobacteroidaceae families. Phylogenetic analysis, phenotypic and physiological characteristics showed a novel strain Sporohalobacter salinus related to Sporohalobacter lortetii, an unique species of genus Sporohalobacter described until now.

Les mucilages pélagiques marins (MPM) sont des phénomènes mondiaux sporadiques et, dans certaines régions, chroniques dans les zones côtières. Ces agrégats gélatineux, enrichis en matière organique et en microorganismes, forment des écosystèmes marins autonomes transitoires allant de 0,5 cm à plusieurs kilomètres de long. Ils correspondent à des étapes évolutives de la neige marine non-sédimentée maintenue dans la zone photique. Durant les dernières décennies une intensification des apparitions des MPM ont été recensées. Ainsi, les MPM sont devenus un sujet de préoccupation pour les populations qui exploitent les ressources côtières et dans le contexte du fonctionnement global de l'écosystème. Malgré l’intérêt scientifique grandissant au cours des dernières décennies au sujet de ces phénomènes, notamment en mer Adriatique, les causes de formation, la dynamique et le rôle respectif des microorganismes dans de tels systèmes restent énigmatiques. La plupart des études réalisées sur les MPM étaient axées uniquement sur les communautés microbiennes eucaryotes par l’utilisation de techniques microscopiques. Or de nos jours, les méthodes moléculaires permettent de se concentrer également sur l'ensemble de la communauté, y compris les procaryotes. Afin d'approfondir notre compréhension de ces phénomènes nous avons étudié un type MPM qui est apparu de manière récurrente et abondante au cours de la dernière décennie dans le sud du Golfe de Gascogne (France), le «Liga». Dans un premier temps, nous avons étudié la formation de ce MPM au travers d’une année complète en suivant la dynamique des communautés archées, bactériennes et eucaryotes par la technique de T-RFLP, technique d’empreinte moléculaire ciblant la petite sous-unité du gène codant pour ARNr. Cette approche a révélé que les communautés microbiennes du Liga étaient différentes des communautés microbiennes marines pour les trois domaines du vivant, et que ces deux communautés étaient gouvernées par des paramètres environnementaux dont la variation était saisonnière. Les archées n’ayant pas pu être détectés dans le Liga, nous nous sommes focalisés sur la structure des communautés bactériennes et eucaryotes au moyen de technologies de séquençage à haut débit. Cette méthode a révélé que le Liga était principalement composé d'espèces marines, même si ces communautés étaient significativement différentes des espèces marines. Dans le Liga, les communautés eucaryotes étaient principalement composées de dinoflagellés, de zooplancton et de cnidaires. Les communautés bactériennes étaient principalement composées d’Alphaproteobacteria et de Gammaproteobacteria. La diversité fonctionnelle du Liga fraîchement formé a été également étudiée pendant les saisons d'apparition de ce phénomène, au printemps et à l'automne. Nous avons ainsi mis en évidence que les communautés microbiennes du Liga avaient potentiellement moins de capacités de résistance au stress et que ces communautés étaient potentiellement plus virulentes que les communautés microbiennes marines. / Marine pelagic mucilage (MPM) is worldwide phenomena occurring sporadically and, in certain regions, episodically in coastal areas. These gelatinous aggregates, enriched in organic matter and microorganisms, form autonomous transitory marine ecosystems ranging from 0.5 cm to several kilometers. They correspond to evolving stages originating from the non-settling early marine snow maintained in the photic zone. During the last decades intensification of MPM events have been noticed. They became a matter of concern both for populations exploiting coastal resources and in the context of global ecosystem functioning. Although increased scientific attention has been paid during last decades to these phenomena in specific areas such the Adriatic Sea, the causes of appearance, the dynamics, and the respective role of microorganisms in such systems remain enigmatic. Most of the studies performed on MPM focused on eukaryotic microbial communities using microscopic techniques. However molecular methods allow nowadays focusing also on the whole community including the prokaryotic part. In order to deeper our understanding of these phenomena we studied a MPM that occurred recurrently and more frequently during the last decade in the south of the Bay of Biscay (France), the “Liga”. In a first step we investigated the formation of this MPM through a complete year by following the dynamics of archaeal, bacterial and eukaryotic communities using T-RFLP fingerprinting targeting the small subunit of rRNA genes. This approach revealed that Liga’s microbial communities where different from marine microbial communities for the three domains of life and that both marine mucilage and marine communities were linked with seasonal patterns. As archaea were not found in the Liga, we focused on bacterial and eukaryotic communities’ structures through high throughput sequencing. The molecular composition revealed that the Liga was mainly composed of marine species although these communities were significantly different from marine species. In the Liga, eukaryotic communities were mainly composed of dinoflagellates, zooplankton and cnidarians species and bacterial communities were mainly composed of Alphaproteobacteria and Gammaproteobacteria. Functional diversity of freshly-formed Liga was targeted during its seasons of apparition, in spring and in autumn. We highlighted that Liga’s microbial communities had less potential capabilities to resist to stress conditions and that these communities were potentially more virulent than marine microbial communities.

Este trabalho investigou a diversidade microbiana e a filogenia de arquéias e bactérias em consórcios anaeróbios metanogênicos, originados de sedimentos estuarinos enriquecidos com pentaclorofenol (PCP) e 2,6-diclorofenol (2,6-DCP). Para tanto foram construídas bibliotecas genômicas e utilizados métodos moleculares independentes de cultivo como a Eletroforese em Gel de Gradiente Desnaturante (DGGE) e o seqüenciamento de segmentos específicos do DNAr 16S microbiano. Os resultados da DGGE permitiram verificar alterações na estrutura das comunidades microbianas, as quais provavelmente ocorreram devido às adversidades ocorridas nos sistemas durante o período de incubação como a entrada de oxigênio nos frascos e o acúmulo de compostos clorados no meio de cultivo, principalmente o 2,6-DCP. A estimativa da diversidade beta, realizada pela comparação dos padrões de bandas da DGGE, também permitiu inferir que as alterações nas composições das comunidades de arquéias e bactérias foram devidas às duas estratégias empregadas para o enriquecimento da microbiota autóctone do estuário estudado, ou seja, a pasteurização/não pasteurização das amostras de sedimentos estuarinos. Os resultados das análises filogenéticas revelaram que as seqüências analisadas dos clones bacterianos foram relacionadas ao grupo das bactérias Gram-positivas com baixo conteúdo de G+C pertencentes à Ordem Clostridiales (100%) do Filo Firmicutes e as das arquéias relacionadas ao grupo das metanogênicas pertencentes às Ordens Methanobacteriales (7,1%), Methanosarcinales (14,3%), Methanomicrobiales (57,1%) e arquéias não identificadas (21,4%) do Filo Euryarchaeota. Em relação às bactérias, alguns clones foram identificados como pertencentes aos gêneros Sedimentibacter, Clostridium e Alkalibacter, os quais são representados por microrganismos que apresentam metabolismo fermentativo e requerem a presença de extrato de levedura para o crescimento. Provavelmente as bactérias analisadas neste trabalho fermentaram a glicose e o piruvato, os quais foram utilizados como doadores de elétrons, com conseqüente produção de lactato, etanol, butirato, acetato, formiato e hidrogênio/gás carbônico, que podem ter sido utilizados por outros grupos de microrganismos no processo global da digestão anaeróbia. Tais bactérias foram importantes para o processo global de degradação dos clorofenóis, pois também utilizaram como doadores de elétrons os produtos parcialmente degradados por outras bactérias fazendo com que não houvesse acúmulo desses compostos no sistema. Enquanto que algumas arquéias foram relacionadas a organismos hidrogenotróficos pertencentes aos gêneros Methanoculleus e Methanocalculus, Methanobacterium, e acetotróficos do gênero Methanosaeta, as quais utilizaram como substratos para a metanogênese os subprodutos da fermentação bacteriana, ou seja, o \'H IND.2\'/\'CO IND.2\', o formiato e o acetato, contribuindo assim para a manutenção do equilíbrio das demais reações ocorridas no sistema. Desta forma, os dados obtidos neste trabalho poderão auxiliar na compreensão do funcionamento de ecossistemas contaminados por compostos clorados, bem como contribuir para o desenvolvimento de novos processos biotecnológicos aplicados aos problemas ambientais. / This work aimed to investigate microbial diversity and phylogeny of archaea and bacteria microrganisms methanogenic in estuarine sediment samples enriched with organic sources under methanogenic and halophlic conditions. The samples were obtained from previous study on anaerobic degradation of pentachlorophenol (PCP) and 2,6-dichlorophenol (2,6-DCP). Microbial studies were done by the molecular methods without cultivation requirement, Denaturing Gradient Del Electrophoresis (DGGE) and specific segments of 16S rDNA sequencing. DGGE-profile showed structure changes in microbial community. Probably, as a consequence of \'O IND.2\' intake and accumulation of chlorinated compounds, mainly 2,6-DCP, in the culture medium. The beta diversity estimation showed that changes in arquaea and bacteria communities probably occurred as a consequence of the two enrichment strategies used for estuarine indigenous microorganisms, pasteurization and non-pasteurization of the estuarine sediments samples. Phylogenetic analysis indicated the presence of gram-positive bacterium with low G+C content related to Clostridiales Order (100%) belonging to Firmicutes Phylum, as well as related to methanogenic archaea from Methanobacteriales (7,1%), Methanosarcinales (14,3%) and Methanomicrobiales (57,1%) Order. Non-identified archaeas from Euryarchaeota Phylum were also found (21,4%). Concerning to bacterias, it was identified clones related to genera Sedimentibacter, Clostridium and Alkalibacter, which have fermentative metabolism and require yeast extract to grow. Probably, bacterial cells analised in this work fermented glucose and pyruvate producing lactate, ethanol, butirate, acetate, formiate and hydrogen/carbon dioxide. All these products could be used for other microbial groups in the global process of anaerobic digestion. The bacterial microorganisms were important to global process of chlorophenol degradation because contributed in the overall process utilizing the products partially degraded by other bacterias and preventing its accumulation in the system. Identified archaeal cells were related to hydrogenotrophs microorganisms of the genera Methanosaeta, Methanoculleus, Methanocalculus and Methanobacterium, which utilized the bacterial fermentation sub-products as acetate, \'H IND.2\'/\'CO IND.2\' and formiate as substrate for the methanogenesis, contributing for the maintenance of the balance of other reactions occurred in the system. The results of this work give advanced knowledge about understanding biotechnological process of contaminated ecosystems by chlorinated compounds. Therefore, it could contribute to development of new biotechnological processes applied to environmental problems.

Les sols constituent le principal réservoir de carbone, avec près de deux fois plus de carbone que le pool atmosphérique. Afin de pouvoir prédire et anticiper le devenir du carbone dans le contexte actuel de changement climatique et de changement d'usage des terres, il apparaît nécessaire de mieux comprendre les processus qui régulent la décomposition des matières organiques dans les sols. Cette thèse se propose donc d'étudier deux types de régulateurs de la dynamique du carbone du sol : les propriétés de l'habitat microbien et celles des communautés microbiennes. En effet, puisque directement affectées par les changements climatiques d'une part, et les changements d'usage des terres et de pratiques culturales d'autre part, l'habitat microbien et les communautés microbiennes apparaissent comme des régulateurs clés de la dynamique du carbone du sol. Des dispositifs expérimentaux permettant de faire varier les propriétés de l'habitat microbien et celles des communautés microbiennes de façon indépendante ou simultanée ont été mis en place. Dans un premier temps, des microcosmes dont la structure du sol a été manipulée afin d'obtenir des gradients de déstructuration, ont été incubés. Dans un second temps ce sont des microcosmes mettant en jeu des gradients de diversité microbienne qui ont été incubés. Enfin, une incubation utilisant les différences naturelles de propriétés de l'habitat microbien et de communautés microbiennes a été mise en place pour tenter de hiérarchiser ces régulateurs de la décomposition des matières organiques du sol. Les résultats obtenus ont mis en évidence que l'activité microbienne de décomposition du carbone organique du sol semble plus contrôlée par les conditions environnementales (comme le pH, la texture et l'approvisionnement en substrat) que par la structure des communautés microbiennes ou leurs capacités métaboliques. En plus de cela, la fonction de minéralisation ne semble être affectée que dans le cas d'une très grande érosion de la biodiversité suggérant la présence d'un effet seuil, et que l'importance de la redondance fonctionnelle n'est pas toujours aussi grande que ce que le suggère de nombreuses études. D'autre part, dans des conditions d'aération suffisante, les mécanismes qui réguleraient la dynamique du carbone organique des sols se passeraient à des échelles très fines. / Soils represent the principal reservoir of carbon with two times as much carbon as is found in the atmospheric pool. In an effort to better predict and anticipate how soil carbon dynamics will be affected by environmental changes and by the evolution of cropping systems, it is necessary to better understand the processes that regulate soil organic matter decomposition. This study aims to investigate two regulatory mechanisms of the soil carbon dynamic: the properties of the microbial habitat and the ones of the microbial communities. Because they are directly affected by the climatic changes and by the rapid evolution of cropping systems these two mechanisms appear to have a key role in the regulation of soil carbon decomposition. Experimental designs were setup allowing the variations, independent or simultaneous, of the properties of microbial habitat and the ones of the microbial communities. First, to assess the relative importance of soil structure, microcosms with different gradient of disaggregation were incubated. Then, to assess the relative importance of diversity erosion, microcosms with microbial diversity gradient were incubated. Finally, using contrasted soils varying in their habitats and their microbial communities properties, we aimed to hierarchize these two carbon decomposition regulatory mechanisms.The obtained results indicate that microbial activity of soil organic carbon decomposition seems to be more controlled by environmental conditions (such as pH, texture and also substrate supply) than by the microbial community structure or metabolic profiles. Then we observed that organic carbon mineralisation was impacted only when the levels of diversity were very low suggesting the existence of a threshold, and that the functional redundancy is maybe not as great as numerous studies suggest. Moreover, our work showed that when conditions of aeration in the pore system are sufficient, mechanisms regulating the dynamic of soil organic carbon take place at fine spatial scales.