Mikrobielle Prozesse spielen eine wichtige Rolle im Sediment von Talsperren und Seen. Demgegenüber stehen nur unzureichende Erkenntnisse über die Zusammensetzung mikrobieller Biozönosen in Sedimenten sowie deren Aktivität zur Verfügung. Das Ziel dieser Studie war die Untersuchung und der Vergleich der Zusammensetzung und der Struktur mikrobieller Gemeinschaften in Sedimenten um eine Abschätzung der mikrobiellen Diversität in Talsperrensedimenten unterschiedlicher Trophie zu erreichen. Durch die Kombination der in dieser Arbeit verwendeten Methoden (Vergleichende 16S rDNA Analyse, Fingerprinttechniken, klassische Methoden) konnte eine Charakterisierung der mikrobiellen Zusammensetzung der obersten 5 cm von den Talsperrensedimenten Neunzehnhain, Muldenberg, Quitzdorf und Saidenbach erzielt werden. Die vergleichende 16S rDNA Analyse offenbarte in 2541 analysierten rekombinanten Klonen 528 verschiedene Sequenztypen, welche zu 293 OTUs zusammengefaßt werden konnten. Obwohl die Gemeinschaften der verschiedenen Talsperren nur schwach auf der Ebene der phylogenetischen Gruppen differierten, konnte durch die Verwendung von Ähnlichkeitsindices gezeigt werden, dass jede Talsperre eine spezifische mikrobielle Sedimentgemeinschaft aufweist. Über 60% aller Klone zeigten Ähnlichkeiten von mehr als 97% zu 16S rDNA-Sequenzen kultivierter Organismen oder phylogenetisch eingeordneten Sequenzen (14 bekannte phylogenetische Gruppen). Alle anderen Klone zeigten hohe Sequenzhomologien zu unidentifizierten, phylogenetisch bisher nicht eingeordneten Bakterien. Diese Bakterien waren mit Anteilen zwischen 19,8% (Muldenberg) und 54,6% (Saidenbach) in den 16S rDNA Bibliotheken repräsentiert. Mittels Fingerprinttechniken (DGGE, T-RFLP, ARISA) konnten komplexe Muster der mikrobiellen Diversität erzeugt werden. Dabei konnten die Ergebnisse der 16S rDNA Analyse bestätigt werden. Durch die verwendeten Methoden konnte eine komplexe mikrobielle Diversität in den Sedimenten aufgedeckt werden und die Ergebnisse weisen darauf hin, dass die mikrobielle Diversität in Sedimenten wesentlich höher ist als bisher angenommen.

The influence of uranyl and sodium nitrate under aerobic and anaerobic conditions on the microbial community structure of a soil sample from the uranium mining waste pile Haberland in Germany was studied by using the 16S rRNA gene retrieval. The results demonstrate a shifting in the bacterial populations depending on the treatment, whereas the archaeal community was changed independently of the treatment. By using the nitrate reductase gene (narG) as a functional marker, it was additionally demonstrated that some of the bacteria stimulated possess the membrane-bound nitrate reductase. In addition, two Arthrobacter strains were isolated from the studied uranium mining waste pile, which tolerate relatively high concentrations of uranium and heavy metals. These strains are able to precipitate lead as lead sulphide (galena) or lead phosphate mineral phase (pyromorphite) depending on their physiological state and to accumulate uranium intracellularly. The results demonstrate a microbial community in the uranium mining waste pile Haberland, which is able to influence the fate of uranium.

O objetivo deste trabalho foi estudar a aplicação de reator anaeróbio serial (RAS) no processamento de vinhaça de cana-de-açúcar para a remoção de matéria orgânica e produção de biogás, submetido ao aumento gradativo de carregamento orgânico. Para tanto, foi utilizado um reator composto por quatro câmaras em série denominadas C1, C2, C3 e C4, com capacidade de 5 L cada, totalizando um volume de 20 L. A primeira etapa do trabalho (Etapa 1) consistiu na operação e monitoramento do reator RAS partindo-se da aplicação da carga orgânica volumétrica (COV) de 0,5 kg DQO m-3.d-1 até a obtenção da COV de 2,5 kg DQO m-3.d-1. Na segunda etapa (Etapa 2) foram operados reatores em batelada sequencial, simulando a operação contínua do reator RAS, visando estudar o efeito do aumento de taxas de carregamento orgânico aplicado. O reator RAS operou sob baixos valores de COVglobal e COALglobal aplicados na Etapa 1, obtendo-se máximas de 2,50 kg DQO m-3.d-1 e 0,167 kg STV m-3.d-1, respectivamente. Os resultados obtidos da Etapa 1 indicam satisfatória remoção global média de matéria orgânica na forma de DQO, superiores a 90 %, propiciada pela adaptação da biomassa ao longo dos reatores sequenciais. O metano apresentou-se como o gás predominante na composição do biogás produzido, com valores acima de 75 %, que confirmam o estabelecimento do sistema metanogênico pleno. Em todos os compartimentos dos reatores sequenciais verificou-se maior abundância relativa para os domínios Archaea e Bacteria os gêneros Methanosaeta, produtores de metano e, T78, responsável pela degradação de açúcares, respectivamente. Principalmente para os reatores C3 e C4, observou-se adaptação de algumas populações para ambos os domínios Bacteria e Archaea indicado pelo menor índice de diversidade e aumento da dominância. O aumento das taxas de carregamento orgânico aplicado ao lodo, estipuladas nos ensaios em batelada na Etapa 2, propiciou o aumento dos valores de carga orgânica aplicada para todos os reatores. Com o aumento do carregamento orgânico aplicado, observa-se o aumento e acúmulo de ácidos orgânicos para todas as fases dos ensaios, principalmente o acético. A produção de metano foi significativa na operação dos reatores em batelada, principalmente em C1. De forma geral, os resultados obtidos demonstraram o bom desempenho do processamento da vinhaça de cana-de açúcar em reator compartimentado tipo RAS para a remoção de matéria orgânica e produção de biogás. / The aim of this study was to investigate the application of an anaerobic serial reactor (ASR) in the sugarcane stillage processing for the removal of organic matter and biogas production, submitted to the gradual increase in organic loading rate. ASR reactor consisted of four reactors operated in series (C1, C2, C3 and C4), each with a reaction volume of 5 L (total volume of 20 L). The first step (Step 1) consisted of reactor operation and monitoring; organic loading rate was gradually increased from 0.5 kg COD m-3d-1 up to 2.5 kg COD m-3 d-1. In the second step (Step 2), sequential batch reactors were operated, simulating the continuous ASR reactor operation, aiming to study the effect of the increase of organic load applied to sludge (OLS). The ASR reactor operated under low values of COVglobal and COALglobal applied in Step 1, obtaining a maximum 2.50 kg COD m-3d-1 and 0.167 kg TVS/m-3d-1, respectively. The results obtained from Step 1 indicated satisfactory removal of the global average organic matter in the form of COD more than 90 %, caused by adaptation of the biomass along the sequential reactors. Methane appeared to be the predominant gas in biogas composition, with values above 75%, which confirmed the establishment of the full methanogenic system. In all compartments of the sequential reactors, there was a higher relative abundance for the domains Archaea and Bacteria of the genera Methanosaeta, methane producers, and T78, which is responsible for degradation of sugars, respectively. Especially for the reactors C3 and C4, the adaptation of some populations for both domains Bacteria and Archaea indicated was observed by the lower diversity index and increase in dominance. The increase of the organic loading rates applied to the sludge, as established in the batch tests, increased the organic load values applied to all reactors. With the increase of organic loading applied, an increase in the accumulation of organic acids, mainly acetic, for all stages and the batch tests. Methane production was significant in the operation of batch reactors, mainly in the C1. In general, the results obtained showed good performance of the processing of the sugarcane stillage in serial reactor (ASR) for the removal of organic matter and biogas production.

A Política Nacional de Resíduos Sólidos, instituída pela Lei Federal 12.305 em agosto de 2010 e regulamentada pelo Decreto 7.404, em 23 de dezembro de 2010, estabeleceu um novo paradigma na gestão e gerenciamento dos resíduos sólidos no Brasil. Entre as mudanças apresentadas, uma das mais relevantes é a que prioriza o não aterramento da fração orgânica e a recuperação energética dos resíduos. Por esse motivo, a busca por novas tecnologias para o tratamento desses resíduos tem sido intensificada nos últimos anos. Nesse contexto, o objetivo deste trabalho busca avaliar o potencial de se realizar estratégias para aclimatação de lodos anaeróbios em possíveis inoculações de biometanizadores alimentados com fração orgânica dos resíduos sólidos urbanos (FORSU). Foram analisados dois diferentes lodos, esses em função da decomposição e estabilização da FORSU por meio de ensaios de biodegradabilidade anaeróbia. O primeiro, lodo ETE (LE), foi coletado de um reator UASB, utilizado para tratamento de esgoto sanitário doméstico, já o segundo, lodo Aterro (LA), foi coletado de uma lagoa de acúmulo de lixiviados em um aterro sanitário desativado. Para os ensaios, utilizaram-se frascos Duran® de 250 mL, incubados a 35°C, considerando-se a co-digestão nas seguintes relações resíduo/inóculo (R0/I0): 1/16, 1/8, 1/4, 1/2 e 1/1 gSTVresíduo.gSTVinóculo-1. As estratégias de aclimatação consistiram em manter o inóculo para as próximas bateladas, bem como, dobrar as relações (R0/I0) com sucessivas alimentações realizadas em três bateladas sequenciais. Para que fossem possíveis o controle e a verificação do processo ao longo das bateladas, foram realizadas análises físico-químicas, microbiológicas, cinéticas e de atividade metanogênica específica (AME). Os inóculos adaptaram-se de imediato à FORSU, tendo sido verificada produção crescente de metano já nos primeiros dias dos testes. Os melhores resultados do potencial bioquímico de metano foram obtidos pelas menores relações (R0/I0) para ambos os inóculos. As maiores relações (R0/I0) influenciaram negativamente o desempenho do processo de biometanização para o inóculo LE, verificada pelas menores taxas máximas de rendimento de metano e maiores fase lag. Para o inóculo LA, ao final do processo de aclimatação foi verificado maior índice de diversidade para o Domínio Archaea, corroborando com os resultados obtidos pela quantidade de micro-organismos metanogênicos. Essas verificações se deram por meio de análises de microscopia óptica e pelos maiores valores obtidos nos testes de AME. Destarte, conclui-se que a estratégia de aclimatação se utilizando das menores relações (R0/I0): 1/16, 1/8 e 1/4 para o inóculo LA, possui o maior potencial, sendo esse na inoculação em maiores relações (R0/I0) na partida de biometanizadores alimentados com fração orgânica dos resíduos sólidos urbanos, logo, garantindo-se efetividade no processo. / The National Policy on Solid Waste, instituted by Federal Law 12,305 in august 2010 and regulated by Decree 7404, on december 23, 2010, established a new paradigm in the management of solid waste in Brazil. Among the changes presented, one of the most relevant is the one that prioritizes the non-grounding of the organic fraction and the energy recovery of the waste. For this reason, the search for new technologies for the treatment of this waste has been intensified in recent years. In this context, the objective of this work is to evaluate the potential of strategies for acclimatization of anaerobic sludge in possible inoculations of biomethanizers fed with organic fraction of municipal solid waste (OFMSW). Two different sludges were analyzed, as a function of the decomposition and stabilization of OFMSW by anaerobic biodegradability tests. The first, ETE sludge (LE), was collected from a UASB reactor, used to treat domestic sanitary sewage, while the second, Aterro sludge (LA) was collected from a pond of leachate accumulation in a deactivated landfill. For the tests, 250 ml Duran® flasks incubated at 35°C were used, considering co-digestion in the following residue/inoculum ratios (R0/I0): 1/16, 1/8, 1/4, 1/2 and 1/1 gSTVresidue.gSTVinoculum-1. The acclimation strategies consisted in maintaining the inoculum for the next batch, as well as doubling the ratios (R0/I0) with successive feedings performed in three sequential batches. In order to control and verify the process throughout the batch, physical-chemical, microbiological, kinetic and specific methanogenic activity (SMA) analyzes were performed. The inocula immediately adapted to OFMSW, and there was a growing production of methane already in the first days of the tests. The best results of the biochemical potential of methane were obtained by the lower ratios (R0/I0) for both inoculum. The higher ratios (R0/I0) negatively influenced the performance of the biomethanization process for the LE inoculum, verified by the lower maximum methane yield rates and higher lag phase. For the inoculum LA, at the end of the acclimation process, a higher diversity index was verified for the Archaea Domain, corroborating with the results obtained by a number of methanogenic microorganisms. These tests were carried out by means of optical microscopy and the highest values obtained in the SMA tests. Therefore, it is concluded that the acclimation strategy using the lowest ratios (R0/I0): 1/16, 1/8 and 1/4 for the inoculum LA, has the highest potential, being inoculated at higher rates (R0/I0) at the start of biomethanizers fed with organic fraction of municipal solid waste, thus, ensuring effectiveness in the process.

Coral diseases were first noted in the 1960s and 1970s and have had major impacts globally on coral reef community structures. In the Caribbean, a major outbreak of white band disease has been considered responsible for the drastic decline of Caribbean Acroporids since the 1970s. In addition to white band disease, another more recently described condition known as rapid tissue loss (RTL) has had major impacts on Acropora cervicornis populations, specifically offshore Broward County Southeast Florida. While these diseases have contributed to the population decline, determining their etiologies has been elusive. Coral diseases have been characterized by shifts in their microbial counterparts within many levels of the coral host. While some coral diseases have had specific pathogens identified, research has not been able to determine pathogens for most. Evidence points toward bacterial causes for many diseases, but due to the complexity of the coral holobiont and the interaction with the environment, elucidating the causes has proven difficult. Many studies have examined the microbiomes of specific diseases and determined some potential pathogens or at least taxa playing important roles in the disease, although none have looked at RTL. Recognizing the local affect of RTL on A. cervicornis, this study set out to gain a baseline understanding of the healthy and RTL affected microbiome of A. cervicornis. 16S rRNA gene sequencing was used to examine the microbiome of completely healthy colonies, healthy regions of diseased colonies, and the disease margin of diseased colonies. Analysis of four microbial diversity metrics revealed marked increases in diversity with respect to declining health states. Additionally, community dissimilarity analysis and analysis of differentially abundant taxa exhibited distinct microbial community structures due to coral health. Several highly abundant (Rickettsiales, Rhodobacteraceae) and a few low abundance (Bdellovibrionales) taxa were identified as primary drivers of the differences. Additionally, Piscirickettsiaceae, a known fish pathogen, was consistently associated with RTL and warrants further investigation. All of the taxa identified with in RTL have been associated with other Acroporid and non-Acroporid diseases throughout the Caribbean and the rest of the world. The consistent IV association of similar taxa for coral diseases around the world, including those found in this study, supports the recent ideas of non-specific primary pathogens. While most disease studies, coral and otherwise, aim to determine a single pathogen for a single disease, this study and others suggest there could be a multitude of organisms responsible for the disease. Therefore understanding the interactions of the coral holobiont and the environment is important to understanding coral disease. While this study reveals significant changes in the bacterial community associated with RTL as well as some potential pathogens, the relationships appear complex and perhaps at a functional level rather than merely taxonomic. Furthermore, this study did not examine viruses, fungi, or protists, which could be possible pathogens. Therefore, to further develop an understanding of RTL and many other coral diseases it will be necessary to consider additional none-bacterial members of the holobiont as well as the bacterial functions and taxa coupled with the roles of environmental factors.

Les compartiments géologiques profonds suscitent un intérêt grandissant dans la communauté scientifique depuis les 50 dernières années. Néanmoins, ces écosystèmes demeurent largement méconnus du fait de leur difficulté d’accès. Le forage profond réalisé par l’ANDRA dans le Bassin parisien en 2008 a offert une opportunité unique de les étudier. Dans ce cadre, cette thèse avait deux objectifs majeurs ; i) caractériser, d’un point de vue microbiologique, quatre formations sédimentaires terrestres triasiques situées entre 1700 et 2000 m de profondeur et ii) étudier les effets combinés des paramètres de température, pression et salinité ainsi que de leur interaction sur l’activité métabolique de procaryotes anaérobies afin de mieux appréhender leur comportement au cours d’un enfouissement géologique.Malgré la recherche de microorganisme par la réalisation d’une gamme de milieux de culture diversifiée, ciblant préférentiellement les types trophiques fréquemment rencontrés en subsurface (méthanogènes, fermentaires, réducteurs de composés soufrés), aucun microorganisme viable et cultivable n’ait été isolé. En parallèle, une approche moléculaire complémentaire, composée (i) de l’étude comparative de l’efficacité de différentes méthodes d’extraction directe d’ADN et (ii) de l’analyse de la diversité bactérienne par la réalisation d’inventaires moléculaires, par DGGE (Denaturing Gel Gradient Electrophoresis) et clonage, a été réalisée sur le coeur des carottes de roches, conservées à pression atmosphérique ou sous pression, dans leurs états initiaux et post-incubation. L’exploration de ces formations sédimentaires profondes a indiqué la présence d’une très faible biomasse et d’une biodiversité microbienne pauvre principalement composée de membres aérobies et mésophiles appartenant au domaine Bacteria. Cette communauté bactérienne inattendue car a priori peu adaptée aux conditions régnant in-situ, également retrouvée dans divers écosystèmes de subsurface ainsi que dans des biotopes extrêmes, pourrait provenir en partie d’une paléo-recharge de l’aquifère du Trias par des eaux froides dérivées de la fonte des glaces formées lors de la dernière glaciation du Pléistocène.Le second objectif a été abordé à travers l’élaboration d’un plan factoriel complet dans le but d’identifier les effets des paramètres sur les activités microbiennes. Ainsi, les activités métaboliques de huit souches microbiennes halophiles et thermo-tolérantes ont été mesurées sous trente conditions distinctes de température (40, 55 et 70°C), pression (1, 90 et 180 bars) et salinité (13, 50, 110, 180 et 260 g.l-1). Toutes les souches originaires d’environnements profonds se sont révélées être au minimum piézo-tolérantes et capables de maintenir leur activité métabolique sous pressions hydrostatiques. Les métabolismes fermentaires (Thermovirga lienii et Halothermothrix orenii) et thiosulfato-réducteurs (Petrotoga mexicana et Thermosipho japonicus) se sont avérés particulièrement bien adaptées, d’un point de vue métabolique, aux hautes pressions, les plus hautes activités ayant été détectées sous pression. Certaines souches ont montré une résistance accrue aux hautes températures sous pression (Petrotoga mexicana). Toutefois une résistance variable à la salinité dans les différentes conditions de température et de pression a été observée pour chacune des souches, suggérant que certains mécanismes de résistance contre la pression osmotique seraient également efficaces pour lutter contre les températures et les pressions hydrostatiques élevées.Ce travail souligne que l’étude des écosystèmes terrestres profonds d’un point de vue microbiologique ne doit pas se restreindre à la recherche et à l’analyse de la diversité présente. L’étude des activités métaboliques de souches de subsurface en conditions profondes ouvre la voie à une meilleure compréhension des rôles joués par les communautés microbiennes en milieu extrême. / Over the past 50 years, the scientific community has shown a growing interest for deep geological compartments. However, these ecosystems remain largely unknown due to their inaccessibility. The aim of the present thesis was double; the first aim was to characterize, from a microbiological perspective, four terrestrial Triassic sedimentary formations located between 1700 and 2000 m depth in the Parisian Basin and collected by the ANDRA during a deep drilling campaign in 2008, and the second aim was to study the combined effects of temperature, pressure and salinity on the metabolic activity of anaerobic prokaryotes in order to predict their reaction to geological burial.Incubations in a large variety of media were carried out in order to stimulate the growth of the main trophic types found in such environments such as methanogens, fermenters and bacteria reducing sulphur compounds, however, no viable and cultivable microorganisms could be isolated. In parallel, a molecular approach was used to i) compare the efficacy of several DNA extractions methods and ii) analyse the bacterial diversity, using DGGE (Denaturing Gel Gradient Electrophoresis) and cloning, present in rock inner cores conserved either at atmospheric pressure or under pressure, in their initial states and following incubations in various media. The genetic exploration of these samples revealed a very low biomass and a poor diversity composed mainly of aerobic and mesophilic members of the Bacteria domain, a priori unadapted to such a deep, hot, saline and anoxic environment. This unexpected microbial community also found in many subsurface ecosystems as well as in extreme ecosystems could have partially originated from a paleorecharge of the Trias aquifer with cold waters coming from the melting of ice formed during the last Pleistocene glaciation.The second objective was to study the combined effects of temperature (40, 55 and 70°C), pressure (1, 90 and 180 bars) and salinity (13, 50, 110, 180 and 260 g.l-1) on the metabolic activity of anaerobic prokaryotes. In order to identify effects of parameters on microbial activities, a complete factorial plan was constructed from the metabolic activities measured for eight halophile and thermo-tolerant bacterial strains exposed to 30 distinct temperature, pressure and salinity conditions. All the strains issued from deep environments were at the least piezo-tolerant and capable of maintaining their activity under hydrostatic pressures. The fermenting (Thermovirga lienii and Halothermothrix orenii) and thiosulfate reducing strains (Petrotoga mexicana and Thermiosipho japonicus) were particularly well adapted, from a metabolic point of view, to high pressures; indeed the highest activities were measured under pressure. Also, several strains (such as Petrotoga mexicana) showed an increased resistance to high temperatures under pressure. However, resistance to an increase in salinity was variable for each strain under the different temperature and pressure conditions. This suggests that the resistance mechanisms for osmotic pressure also enable resistance to high temperatures and hydrostatic pressures.This work underlines that the microbiological characterization of deep terrestrial ecosystems must not be limited to the search and analyses of the existing diversity. Moreover, such upstream studies of the metabolic activities of subsurface bacterial strains in deep terrestrial conditions are a necessary beginning towards understanding the role of microbial communities in extreme environments.

Xylanases have an important role as biocatalysts in different agroindustrial processes, such as saccharification of plant residues for the production of ethanol and bleaching of wood pulp for the production of pulp. On an industrial scale, these processes may require extreme pH or temperature, which require enzymes compatible with these conditions. Most enzymes available or showing commercial potential is synthesized by fungi or bacteria belonging to a few genera. Evaluation of rare isolation bacteria is an important strategy to expand the diversity of xylanases and their potential in terms of activity, stability and technological application. The aim of this study was to evaluate a collection of common and rare isolation soil bacteria regarding to xylanase activity and characterize the stability conditions of temperature and pH. The analyzed collection consists of 120 isolates with representatives from six phyla that were subjected to screening for xylanase activity in pure cultures and in the extracellular proteic extract (EPE). The ratio between the halos diameters of xylan hydrolysis and in the colonies on solid medium (ratio H:C), incubated at 30 ° C for up to 14 days, was used for the evaluation of cultures as a selection criteria. The effects of different sources of variation in the bacteria isolation stage (original soil, culture medium, solidifying agent, inoculum dilution and plating method and incubation time to colony appearance) and the group of bacterial isolation (rare or common) on the frequency of isolates with high xylanase activity were evaluated based on the ratio H:C. EPEs were obtained in liquid media containing xylan inoculated with the eleven isolates with highest ratios H:C. The extracts were evaluated for the specific xylanase activity at 50 °C for 1 h. Extracts of the three isolates with the highest potential for activity under this condition were evaluated for optimum activity, stability, activity at 60 oC and at pH 4.0, 5.5 and 8.0. Twenty-two isolates (25%), including eight from rare isolation, showed xylanase activity under the conditions evaluated, and found a high variability (> 230%) between these isolates. No isolation factor or rare or common isolation condition were associated with efficiency of xylanase activity in solid medium. High variability in specific xylanase activity in EPEs was also found among isolates (1500%), highlighting two rare isolates (TC119 and TC21), from Alfaproteobacteria class, and one common isolate (TC99), from Ralstoniaceae family. Extracellular xylanases from TC21 and TC119 showed high relative activity at temperatures up to 70 oC and were insensitive to pH in the range 4.0 to 8.0; however, TC99 isolate showed optimum temperature at 40 °C and low stability to temperature and pH. Extracellular xylanases from TC119 showed no cellulolytic activity. Rare isolation soil bacteria show high potential as a source of extracellular xylanases adapted to extreme pH and temperature conditions, which are required in agroindustrial processes. / As xilanases apresentam papel relevante como biocatalisadores de diferentes processos agroindustriais, como a sacarificação de resíduos vegetais para a produção de etanol e o clareamento de polpas de madeira para a produção de celulose. Em escala industrial, estes processos podem requerer pH e, ou, temperatura extremos, os quais demandam enzimas compatíveis com estas condições. A maioria das enzimas disponíveis ou com potencial comercial é sintetizada de fungos ou de bactérias pertencentes a poucos gêneros. A avaliação de bactérias de isolamento raro constitui-se em estratégia importante para ampliar a diversidade de xilanases e suas potencialidades em termos de atividade, estabilidade e aplicação tecnológica. O objetivo deste trabalho foi avaliar uma coleção de bactérias do solo de isolamento comum e raro quanto à atividade de xilanases e caracterizar estas enzimas quanto à estabilidade a condições contrastantes de temperatura e pH. A coleção analisada é composta de 120 isolados com representantes de seis filos e foi submetida à seleção quanto à atividade de xilanases em culturas puras e no extrato proteico extracelular (EPE). Para a avaliação das culturas, utilizou-se como critério de seleção a relação entre os diâmetros de halos de hidrólise de xilana e das colônias em meio sólido (relação H:C), incubado a 30 oC por até 14 dias. Os efeitos de diferentes fontes de variação da fase de isolamento destas bactérias (solo de origem, meio de cultura, agente solidificante, diluição do inóculo e método de plaqueamento e tempo de incubação até surgimento de colônias) e do grupo de isolamento bacteriano (raro ou comum) sobre a frequência de isolados com alta atividade de xilanase foram avaliados com base na relação H:C. EPEs foram obtidos em meios líquidos contendo xilana inoculados com os onze isolados com maiores relações H:C. Os extratos foram avaliados quanto à atividade específica de xilanases a 50 oC, por 1 h. Os extratos dos três isolados com maior potencial de atividade sob esta condição foram avaliados quanto à temperatura ótima de atividade, estabilidade de atividade a 60 oC e a valores de pH 4,0, 5,5 e 8,0. Vinte e dois isolados (25%), incluindo oito de isolamento raro, apresentaram atividade de xilanase nas condições avaliadas, sendo encontrada uma alta variabilidade (>230%) entre estes isolados. Nenhum fator de isolamento ou a condição de isolado raro ou comum foram associadas à eficiência de atividade de xilanases em meio sólido. Alta variabilidade de atividade específica de xilanases nos EPEs também foi encontrada entre os isolados (1500%), com destaque para dois de cultivo raro (TC119 e TC21), da classe Alfaproteobacteria, e um de cultivo comum (TC99), da família Ralstoniaceae. As xilanases extracelulares de TC119 e TC21 apresentaram elevada atividade relativa em temperaturas de até 70 oC e foram pouco sensíveis ao pH na faixa de 4,0 a 8,0; entretanto, as de TC99 apresentaram temperatura ótima de 40 oC, baixa estabilidade a temperatura e pH. Xilanases extracelulares de TC119 não apresentaram atividade celulolítica. Bactérias do solo de isolamento raro apresentam alto potencial como fonte de xilanases extracelulares adaptadas a condições extremas de pH e temperatura requeridas em processos agroindustriais.

Altos fluxos positivos de metano para a atmosfera foram detectados na região amazônica. O gás metano é o segundo mais importante gás de efeito estufa e micro-organismos pertencentes ao Domínio Archaea são responsáveis pela produção de aproximadamente 70% do metano emitido para a atmosfera anualmente. O objetivo deste trabalho foi caracterizar a diversidade de arqueias em sedimentos dos rios Floresta e Madeira através de técnicas moleculares e do cultivo de arqueias metanogênicas. A maior parte das sequências obtidas nas duas bibliotecas pertence ao domínio Crenarchaeota, algumas com similaridade menor que 97% às sequências depositadas nos bancos de dados, revelando a existência de grupos ainda não descritos na literatura. Nos enriquecimentos do sedimento do rio Madeira detectaram-se células pertencentes às famílias Methanosarcinaceae e Methanobacteriaceae pelo emprego de sondas fluorescentes de RNA. Culturas dos gêneros Methanosarcina e Methanobacterium foram estabelecidas em laboratório. A grande diversidade de arqueias não cultivadas encontrada vem reforçar a necessidade de se estudar esse grupo, especialmente sua fisiologia e, consequentemente, seu papel ecológico nos diversos ambientes em que são encontrados. / High positive fluxes of methane to the atmosphere have been detected in the Amazonian region. Methane is the second most important greenhouse gas and microorganisms belonging to the Archaea domain are responsible for approximately 70% of the total methane emitted to the atmosphere annually. The objective of this work was to characterize the Archaea diversity in two sites at Madeira and Floresta rivers sediments using molecular techniques and the culturing of methanogenic archaea. Most sequences obtained in the libraries from both rivers belonged to the Crenarchaeota domain, and around half of them presented less than 97% of similarity to sequences available in databases, revealing the existence of new archaea groups yet to be described in the literature. Cells belonging to the Methanosarcinaceae and Methanobacteriaceae families were detected in the enrichment cultures from Madeira River through the use of RNA fluorescent probes. Strains of Methanosarcina sp. and Methanobacterium sp. are being maintained under laboratory conditions. The great diversity of uncultured Archaea found emphasizes the need to study this group, mainly its physiology and, consequently, its role in the diverse environments they occupy.

The influence of uranyl and sodium nitrate under aerobic and anaerobic conditions on the microbial community structure of a soil sample from the uranium mining waste pile Haberland in Germany was studied by using the 16S rRNA gene retrieval. The results demonstrate a shifting in the bacterial populations depending on the treatment, whereas the archaeal community was changed independently of the treatment. By using the nitrate reductase gene (narG) as a functional marker, it was additionally demonstrated that some of the bacteria stimulated possess the membrane-bound nitrate reductase. In addition, two Arthrobacter strains were isolated from the studied uranium mining waste pile, which tolerate relatively high concentrations of uranium and heavy metals. These strains are able to precipitate lead as lead sulphide (galena) or lead phosphate mineral phase (pyromorphite) depending on their physiological state and to accumulate uranium intracellularly. The results demonstrate a microbial community in the uranium mining waste pile Haberland, which is able to influence the fate of uranium.

Mikrobielle Prozesse spielen eine wichtige Rolle im Sediment von Talsperren und Seen. Demgegenüber stehen nur unzureichende Erkenntnisse über die Zusammensetzung mikrobieller Biozönosen in Sedimenten sowie deren Aktivität zur Verfügung. Das Ziel dieser Studie war die Untersuchung und der Vergleich der Zusammensetzung und der Struktur mikrobieller Gemeinschaften in Sedimenten um eine Abschätzung der mikrobiellen Diversität in Talsperrensedimenten unterschiedlicher Trophie zu erreichen. Durch die Kombination der in dieser Arbeit verwendeten Methoden (Vergleichende 16S rDNA Analyse, Fingerprinttechniken, klassische Methoden) konnte eine Charakterisierung der mikrobiellen Zusammensetzung der obersten 5 cm von den Talsperrensedimenten Neunzehnhain, Muldenberg, Quitzdorf und Saidenbach erzielt werden. Die vergleichende 16S rDNA Analyse offenbarte in 2541 analysierten rekombinanten Klonen 528 verschiedene Sequenztypen, welche zu 293 OTUs zusammengefaßt werden konnten. Obwohl die Gemeinschaften der verschiedenen Talsperren nur schwach auf der Ebene der phylogenetischen Gruppen differierten, konnte durch die Verwendung von Ähnlichkeitsindices gezeigt werden, dass jede Talsperre eine spezifische mikrobielle Sedimentgemeinschaft aufweist. Über 60% aller Klone zeigten Ähnlichkeiten von mehr als 97% zu 16S rDNA-Sequenzen kultivierter Organismen oder phylogenetisch eingeordneten Sequenzen (14 bekannte phylogenetische Gruppen). Alle anderen Klone zeigten hohe Sequenzhomologien zu unidentifizierten, phylogenetisch bisher nicht eingeordneten Bakterien. Diese Bakterien waren mit Anteilen zwischen 19,8% (Muldenberg) und 54,6% (Saidenbach) in den 16S rDNA Bibliotheken repräsentiert. Mittels Fingerprinttechniken (DGGE, T-RFLP, ARISA) konnten komplexe Muster der mikrobiellen Diversität erzeugt werden. Dabei konnten die Ergebnisse der 16S rDNA Analyse bestätigt werden. Durch die verwendeten Methoden konnte eine komplexe mikrobielle Diversität in den Sedimenten aufgedeckt werden und die Ergebnisse weisen darauf hin, dass die mikrobielle Diversität in Sedimenten wesentlich höher ist als bisher angenommen.