Les procédés anaérobies de traitement de déchets apparaissent clairement pouvoir répondre à l'enjeu socio-économique actuel que représente la valorisation énergétique de la fraction organique contenue dans les déchets ménagers. En effet, les processus de dégradation anaérobies font intervenir en cascade, différentes réactions et populations de micro-organismes permettant de transformer la matière organique en biogaz riche en méthane. Une bonne connaissance des effets des paramètres opérationnels sur l'orientation des métabolismes s'avère ainsi nécessaire à l'émergence de solutions permettant d'optimiser ces procédés. Ceci est notamment le cas pour la dernière étape, appelée méthanogenèse. Dans ce contexte, l'approche isotopique reposant sur la mesure de la composition isotopique (13C/12C) du méthane et du dioxyde de carbone, devrait pouvoir répondre à cet objectif en permettant l'identification des métabolismes à l'origine de la production du méthane. La transposabilité à l'étude de la digestion anaérobie des déchets de cette approche isotopique déjà utilisée dans les écosystèmes naturels, a tout d'abord été vérifiée expérimentalement. Les effets de certains paramètres opérationnels connus pour avoir un impact fort sur le processus de digestion anaérobie, tels que la température et la concentration en azote ammoniacal, ont ensuite été étudiés. Il a été mis en évidence qu'en condition thermophile, la méthanogenèse acétoclaste observée en condition mésophile, était remplacée par une oxydation syntrophique de l'acétate lors de la digestion anaérobie des déchets ménagers. Des expériences sur acétate ont montré que cet effet sur les voies métaboliques n'était toutefois pas systématique et pourrait ne pas être dû à un effet direct d'une augmentation de la température, mais plutôt à l'accroissement de la concentration en ammoniaque qui en résulte. D'autres expériences ont clairement établi qu'une augmentation de la concentration en azote ammoniacal conduisait également à la mise en place de l'oxydation syntrophique de l'acétate. Le couplage de l'approche isotopique avec des analyses microbiologiques a révélé que cette réaction d'oxydation syntrophique de l'acétate, à haute concentration en azote ammoniacal, pouvait s'établir telle que déjà décrite, par la mise en place d'une relation symbiotique bactéries/archées hydrogénotrophes strictes, mais également de manière différente en impliquant des membres de la famille Methanosarcinaceae qui pourraient réaliser seuls les deux étapes de la réaction (oxydation et méthanogenèse hydrogénotrophe). L'application de l'approche isotopique a également permis de mettre en évidence, lors d'une expérience visant à simuler la recirculation de différents effluents au sein d'une installation de stockage de déchets bioactive, l'influence de la nature de l'effluent sur l'orientation des métabolismes méthanogènes. Enfin, l'influence de la proportion de déchets verts, lors de la co-digestion biodéchets / déchets verts, sur la concentration en ions ammonium libérés ainsi que sur l'orientation du métabolisme en résultant, a été étudiée. Les potentialités d'une utilisation de l'approche isotopique sur site ont également été investiguées au travers d'une campagne de mesures sur une installation de stockage de déchets non dangereux. / Anaerobic waste treatment processes are clearly part of the answer to a current important socio-economic issue in waste management: energy production from the organic fraction of municipal solid waste. The anaerobic digestion of municipal solid waste is a complex process involving numerous reactions and microorganism communities. At the end of the degradation process, some biogas with a particularly high methane content is produced. A detailed knowledge on how operational parameters affect metabolism orientations is required to optimize these treatment processes. This is in particular the case for the last degradation reaction called methanogenesis. In this context, an isotopic approach based on isotopic composition measurements (13C/12C) for methane and carbon dioxide can provide some clues with regard to this objective. Indeed, this methodology enables the determination of the methanogenic pathways by which methane is produced.Transferability of the isotopic approach used for natural ecosystems to the field of anaerobic digestion of municipal solid waste was first experimentally verified. In a second time, the effects of some operational parameters known to strongly impact the anaerobic digestion process, such as temperature and ammonia concentration, were studied. During anaerobic digestion of reconstituted municipal solid waste in thermophilic conditions, it was shown that aceticlastic methanogenesis (occurring in mesophilic conditions) was replaced by a syntrophic acetate oxidation reaction. Additional experiments using acetate as sole substrate were performed and showed that this effect on the metabolic pathways was not systematic. Consequently, it cannot be due to a direct effect of the temperature increase. It could rather be explained by the induced and indirect increase in ammonia concentration. Additional experiments clearly demonstrated that an increase in ammonia concentration led to the establishment of a syntrophic acetate oxidation reaction. The isotopic approach was combined with microbiological analyses and showed that the syntrophic acetate oxidation reaction occurring at high ammonia concentration during acetate incubations could have been performed through a syntrophic relationship between bacteria and strict hydrogenotrophic archaea, as previously described in the literature. Interestingly, the syntrophic acetate oxidation could also have occurred using a different pathway relying on members of the Methanosarcinaceae family putatively able to perform the two steps of the reaction (oxidation and hydrogenotrophic methanogenesis). In addition, the implementation of the isotopic approach during an experiment designed to simulate a landfill bioreactor evidenced the influence of the effluent's nature on the methanogenesis metabolism orientation. The influence of green waste proportion during the co-digestion of biowaste / green waste mixtures on resulting ammonia concentrations and methanogenesis pathways was also studied through dedicated experiments. Finally, the potential of the isotopic approach for landfill-scale application was investigated through a measurement campaign on a landfill site.

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Notre étude porte sur la preuve de concept de contrôle électrochimique de la méthanogénèse, métabolisme clé de la digestion anaérobie et de la bioremédiation des eaux, en exploitant le principe des piles à combustible microbiennes. Une première partie bibliographique vise à décrire les mécanismes de la méthanogénèse dans le contexte de l'auto-épuration des eaux et de production naturelle de gaz à effet de serre (GES). Les technologies de pile à combustibles microbiennes y sont traitées. Une analyse critique des études sur le contrôle électrochimique de la méthanogénèse permet de dimensionner un montage expérimental dédié à la quantification des GES en cultures biologiques électro-stimulées. Sa conception, sa validation ainsi que les méthodes de mise en culture sont décrites dans une seconde partie. Une série de cultures préliminaires sur des boues digérées anaérobies de station d'épuration permettent d'identifier et fixer les paramètres expérimentaux. Dans une troisième partie, une étude expérimentale fait la preuve de concept de contrôle électrochimique de la méthanogénèse avec une diminution significative de 33% en CH4 (tension de +300 mV vs Ag/AgCl) par rapport à la méthanogénèse naturelle non stimulée. Toutefois, la stimulation contribue à multiplier par 10 la production de CO2. Ce constat amène la problématique supplémentaire d'impact sur l'effet de serre des cultures étudiées. Nous allons donc plus loin que l'objectif initial en nous intéressant à l'empreinte carbone générée par l'ensemble des GES. Le traitement électrochimique, outre la diminution du CH4 produit, permet de diminuer la contribution à l'effet de serre de 15% des cultures électro-stimulées / Our study focuses on the proof of concept of electrochemical control of methanogenesis, key metabolism of anaerobic digestion and water bioremediation, using the principle of microbial fuel cells. A first bibliographic section aims to describe the mechanisms of methanogenesis in the context of self-purification of water and natural production of greenhouse gases (GHG). Microbial fuel cell technologies are addressed. A critical analysis of the studies dealing with electrochemical control of methanogenesis makes it possible to size an experimental setup dedicated to quantification of GHGs in electro-stimulated biological cultures. Its design, validation and methods of cultivation are described in a second part. A series of preliminary cultures on anaerobic digested sewage sludge make it possible to identify and set the experimental parameters. In a third part, an experimental study proves the concept of electrochemical control of methanogenesis with a significant decrease of 33% in CH4 (voltage of +300 mV vs Ag/AgCl) compared to natural unstimulated methanogenesis. However, stimulation contributes to a 10-fold increase in CO2 production. This observation leads to the additional problem of impact on the greenhouse effect of the cultures studied. We go further than the initial objective by looking at the carbon footprint generated by all GHGs. The electrochemical treatment, in addition to the reduction of CH4 produced, makes it possible to reduce the contribution to the greenhouse effect of 15% of electro-stimulated cultures

Energy sources usually used are fossil fuels, which are recursosexauríveis and combustion gases responsible for environmental damage generates. For these reasons, it is necessary to identify alternative energy sources that produce less negative impacts. In this scenario, the hydrogen arises as an alternative source of energy, since it is renewable and there's only generation combustion of oxygen and water, and therefore considered a clean fuel. Similarly, the methane is important in decentralised energy generation and contributes to increase the viability of implementing waste treatment processes. The fermentation is a biological process sustainably to produce hydrogen and methane, because you can use as many different types of agro-industrial waste substrate rich in carbs, minimizing the problems caused by the improper disposal of these materials, which can be harmful to the environment. On the above, the objective of this work was to apply agro-industrial residue of coconut processing in anaerobic reactors for biological production of hydrogen and methane, studying the acidogenic phase coupling with metanogênica serial reactors (two-phase system). For development, were used two anaerobic Fluidised bed reactors (RALF) in series, the first being for the biological production of hydrogen from the agro-industrial residue and the second for reusing the effluent of acidogênico first RALF, aiming at the production of methane. The reactors were operated under progressive increase of organic loading rate (TCO), varying the hydraulic detention time (TDH) of operation. For adhesion of microorganisms, employed as materials supports the expanded clay in the reactor for production of hydrogen and the shells of sururu on methane production reactor. It was found that the generation of hydrogen and methane in two-phase system. Higher income and percentage were obtained on biogas in the TDH of 2:00, corresponding to the values of 2.45 mol H2/mol glucose and 33.82%. Already the largest hydrogen production occurred in the TDH of 1:00, it was 0.57 L/h/l. for the production, income and the percentage in methane biogas, highest values were observed in the TDH of 12:00 am such values corresponded, respectively, to 1.56 L/h/L, 0.09 L CH4/g cod and 38.60%. During the operation of the reactors, were observed in all stages, the presence of acetic acid, butyric acid, propionic acid, caproic acid and ethanol. Compared to other systems of anaerobic digestion, two-phase system used in this work showed satisfactory performance parameters. / Fundação de Amparo a Pesquisa do Estado de Alagoas / As fontes energéticas usualmente utilizadas são os combustíveis fósseis, que são recursosexauríveis e cuja combustão gera gases responsáveis por danos ambientais. Por esses motivos, faz-se necessário identificar fontes de energias alternativas que produzam menos impactos negativos. Nesse cenário, o hidrogênio surge como uma fonte alternativa de energia, já que é de origem renovável e na sua combustão há a geração apenas de oxigênio e água, sendo, portanto, considerado um combustível limpo. Similarmente, o metano é importante na geração descentralizada de energia e contribui para aumentar a viabilidade da implementação de processos de tratamentos de resíduos. O processo biológico fermentativo é uma forma sustentável de produzir hidrogênio e metano, pois pode utilizar como substrato diversos tipos de resíduos agroindustriais ricos em carboidratos, minimizando os problemas causados pelo descarte inadequado desses materiais, os quais podem ser nocivos ao meio ambiente. Diante do exposto, o objetivo deste trabalho foi aplicar o resíduo agroindustrial do processamento de côco em reatores anaeróbios para a produção biológica de hidrogênio e metano, estudando o acoplamento da fase acidogênica com a metanogênica em reatores em série (sistema de duas fases). Para o desenvolvimento, foram usados dois reatores anaeróbios de leito fluidificado (RALF) em série, sendo o primeiro para a produção biológica de hidrogênio a partir do resíduo agroindustrial e o segundo para reaproveitar o efluente acidogênico do primeiro RALF, objetivando a produção de metano. Os reatores foram operados sob aumento progressivo da taxa de carregamento orgânico (TCO), variando o tempo de detenção hidráulica (TDH) de operação. Para aderência dos microrganismos, empregaram-se como materiais suportes a argila expandida no reator para produção de hidrogênio e as conchas de sururu no reator de produção de metano. Verificou-se a geração de hidrogênio e metano no sistema de duas fases. Foram obtidos maiores rendimento e porcentagem no biogás de hidrogênio no TDH de 2 h, correspondendo aos valores de 2,45 mol H2/mol glicose e 33,82%. Já a maior produção de hidrogênio ocorreu no TDH de 1 h, que foi de 0,57 L/h/L. Para a produção, o rendimento e a porcentagem no biogás de metano, foram observados maiores valores no TDH de 24 h. Tais valores corresponderam, respectivamente, a 1,56 L/h/L, 0,09 L CH4/g DQO e 38,60%. Durante a operação dos reatores, foram observadas, em todas as fases, as presenças de ácido acético, ácido butírico, ácido propiônico, ácido capróico e etanol. Em comparação a outros sistemas de digestão anaeróbia, o sistema de duas fases usado no presente trabalho apresentou parâmetros de desempenho satisfatórios.

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