A digestão anaeróbia tem se apresentado como um processo de grande interesse sob a ótica da potencial produção de energia renovável (H2 e CH4), considerando-se a ampla variedade de compostos orgânicos que podem ser utilizados. Neste estudo desejou-se avançar na compreensão do sistema de reatores anaeróbios de duas fases (acidogênico seguido de metanogênico) operados em condições termofílicas (55°C), alimentados com melaço da cana-de-açúcar, subproduto da indústria sucroalcooleira. Os experimentos foram conduzidos em reatores anaeróbios de leito fixo estruturado com fluxo ascendente e o melaço foi diluído com água de abastecimento, para adequação da concentração aos processos de tratamento de águas residuárias. Na 1ª Etapa dois reatores acidogênicos foram operados em paralelo para avaliar diferentes formas de inoculação e meios suportes, a fim de manter a produção continua e estável de hidrogênio. Para isso foram aplicadas diferentes cargas orgânicas (2,5, 5 e 10 gDQO.L-1) que resultam em COV de 30, 60 e 120 g.DQO.Lreator1.dia-1, com TDH fixo de 2 horas. A expressão do gene hidrogenase foi detectado em ambos os reatores, mas em maior proporção no reator inoculado com lodo de reator UASB e usando como material suporte a espuma de poliuretano. Sequencialmente a este reator, foi acoplado um reator metanogênico, alimentado com efluente do reator acidogênico, estabilizado nas condições apresentadas, e operado com COV crescentes de 1, 2, 5, 7, 14, 17 e 26,5 gDQO.Lreator-1.dia-1 e consequente diminuição do TDH de 240, 96, 48, 32, 24, 16 e 12 horas. O reator acidogênico na 2ª etapa foi operado por 417 dias consecutivos e COV de 120 g.DQO.Lreator1.dia-1, produzindo hidrogênio continuamente, alcançado valores de produção bruta de H2 de 7,60 LH2.dia-1. O reator metanogênico foi operado por 251 dias consecutivos, produzindo metano e alcançado valores de produção bruta de CH4 de 5,90 LCH4.dia-1. A eficiência de remoção de DQO do sistema de reatores foi de aproximadamente 90%, com contribuição aproximadamente de 10% para o reator acidogênico e contribuição aproximadamente de 80% para o reator metanogênico. O reator acidogênico alcançou rendimento de produção de hidrogênio por kg de melaço aplicado de 392 LH2.kgmelaço-1 e o reator metanogênico de 387 LCH4.kgmelaço-1. Para finalidade de comparações e aplicabilidade, o ganho energético global do sistema de reatores de duas fases foi de aproximadamente 5,7 kWh.kgmelaço-1 (1,4 kWh.kgmelaço-1 para o reator acidogênico e 4,3 kWh.kgmelaço-1 para o reator metanogênico). A produção continua de H2 obtida neste estudo está relacionada à associação das vias dos ácidos produtores de hidrogênio já consolidados pela literatura pertinente (acético e butírico) e pela produção de hidrogênio pela rota do ácido lático, devido a associação entre as comunidades de microrganismos estabelecidas no reator. O sequenciamento massivo MiSeq mostrou a seleção de diversos gêneros de microrganismos com redundância funcional e pertencentes principalmente aos Filos Firmicutes, Proteobacteria e Thermotogae, tais como Clostridium sensu stricto, Thermohydrogenium, Thermoanaerobacterium e Cellulosibacter (Firmicutes); Pseudomonas, Enterobacter, Shewanella e Petrobacter (Proteobacteria) e Fervidobacterium (Thermotogae). Microrganismos produtores de ácido lático também foram selecionados tais como: Lactobacillus, Leuconostoc, Sporolactobacillus e Trichococcus. Dos pontos de vista científico e tecnológico este estudo deu mais um passo para a compreensão dos bioprocessos envolvidos nos sistemas anaeróbios em dois estágios produzindo H2 e CH4 continuamente por longo período de tempo. / Anaerobic digestion has shown as an interesting process for renewable energy production (H2 and CH4), for a wide variety of organic compounds (carbon source). This study aimed to advance the understanding of a two-stage process anaerobic system (acidogenic bioreactor followed by methanogenic bioreactor) under thermophilic condition (55°C) fed with molasses, a sugarcane industry by-product. The experiments were conducted at up-flow structured bed reactors and sugarcane molasses was diluted with tap water, to adjust the concentration to the wastewater treatment. At first stage two acidogenic reactors were operated in parallel to evaluate different source of inocula and support bed, to obtain continuous and stable hydrogen production. It was applied 2.5, 5 and 10 gCOD.L-1 resulting in OLR of 30, 60 and 120 g.COD.Lreactor-1.day-1, with HRT fixed at 2 hours of hydrogenase gene was detected in both reactors but with higher number of copies of the gene in the reactor that showed higher hydrogen production: the reactor sed with sludge of UASB reactor and using polyurethane foam as support material. To this reactor was coupled a methanogenic reactor fed with effluent from acidogenic reactor and operated with increasing OLR (1, 2, 5, 7, 14, 17 e 26,5 gCOD.Lreactor-1.day-1) decreasing the HRT (240, 96, 48, 32, 24, 16 and 12 hours). The acidogenic reactor was operated during 471 days with OLR of 120 g.COD.Lreactor-1.day-1, with HRT fixed at 2 hours, with continuous hydrogen production with a gross production of 7.60 LH2.day-1. The methanogenic reactor was operated for 251 days, with continuous methane production of up to 5.90LCH4.day-1. The COD removal efficiency using the two-stage system was approximately 90% , with 10% contribution by the acidogenic reactor and 80% contribution by the methanogenic reactor. The acidogenic reactor achieved hydrogen yield per kg of applied molasses equal to 392 LH2.kgmolases-1. The methanogenic reactor achieved methane yield per kg of applied molasses equal to 387 LCH4.kgmolasses-1. For comparison and applicability purposes, the overall energy yield using the two stage reactor system was approximately 5.7 kWh.kgmolasses-1 (Acidogenic reactor 1.4 kWh.kgmolasses-1 and Methanogenic reactor 4.3 kWh.kgmolasses-1). The continuous production of H2 obtained in this study is related to the association of the hydrogen producer acids pathway established by the relevant literature (acetic and butyric) and the hydrogen production by the lactic acid pathway due to the microorganisms association established in the reactor. Metagenomic analysis by MiSeq Plataform revealed that hydrogen production was due the selection of microorganisms with functional redundancy mainly of Phyla Firmicutes, Proteobacteria and Thermotogae, such as Clostridium sensu stricto, Thermohydrogenium, Thermoanaerobacterium, Cellulosibacter (Firmicutes); Pseudomonas, Enterobacter, Shewanella and Petrobacter (Proteobacteria) and Fervidobacterium (Thermotogae). Genera of acid latic producers, such as Lactobacillus, Leuconostoc, Sporolactobacillus and Trichococcus, were also selected. From the scientific and technological point of view this study has taken another step towards the understanding of bioprocesses involving two stage anaerobic systems for a long term continuous production of H2 and CH4.

Municipal wastewater treatment plants (WWTP) are important infrastructural components in a society and also important for sustainability. In a WWTP the most common treatment configuration is mechanical, biological, and chemical treatment of the wastewater. The treatments reduce nutrients and organic matter before the water is transferred to the recipient. During wastewater treatment, sludge is produced and then stabilized in an anaerobic digestion (AD) process.  The Käppala Association operates the third largest WWTP in Sweden. In the future the Käppala plant is expecting an increased number of connected households and also stricter sludge hygienization regulations. This implies that current strategies for the WWTP have to be developed. One idea has been to use a thermophilic AD process instead of mesophilic AD.Thermophilic AD has previously been shown to have higher capacities, lower hydraulic retention times (HRT) and increased pathogenic destruction compared to their mesophilic counterpart. A common negative aspect for a thermophilic process has been process instability.  In this study a 5 m3 pilot plant rented from Research Institutes of Sweden (RISE) was used to evaluate maximal organic loading rate (OLR) for a stable thermophilic AD process using mixed sludge as substrate. Four HRT were chosen, and each HRT was maintained for 3 retention times. Laboratory analyzes of the raw and digested sludge and on-line monitoring were performed regularly to collect information about process stability and efficiency. The pilot plant was controlled through a surveillance system where operating parameters were introduced. The main objectives of this study were to investigate how an increase of OLR affected pH, alkalinity, and volatile fatty acid (VFA) content and also how the alkalinity and VFA affected the process stability. Gas composition, gas production, degree of digestion (DD) and foaming were also investigated throughout this study. Results showed that VFA initially increased, and alkalinity decreased when every OLR increase occurred. The VFA and alkalinity returned to lower values after the process was given time to recover during constant OLR. The OLR increase caused slight variations in process pH but not enough for process failure. Increased OLR did not seem to have a large impact on the DD since it seemed to remain between 40-50% throughout the entire study. Composition monitoring showed an increased hydrogen sulfide content in the gas as a consequence of increased OLR. An OLR increase also lead to a volumetric gas production (VGP) and volumetric methane production (VMP) increase while efficiency of the AD process was seen to decrease when evaluating specific gas production (SGP) and specific methane production (SMP). The conclusion of the study was that a stable thermophilic AD process using mixed sludge as substrate could be operated with an OLR of 6.55 ± 0.06 kg VS m-3d-1and a HRT of 7 days. Maximal OLR for a stable thermophilic AD process was never achieved due to the process remaining relatively stable the entire experimental period. The process recovered from signs of instability during all HRT-transition times indicating it can maintain all investigated OLR. / Kommunala avloppsreningsverk är en viktig infrastruktur i ett samhälle och även viktigt för hållbarheten. I ett avloppsreningsverk renas spillvatten genom mekaniska, biologiska och kemiska behandlingar processteg. Behandlingarna minskar näringsämnen och organiskt material innan vattnet rinner ut i recipienten. Vid rening av avloppsvatten erhålls slam. Slammet kan användas som substrat för en anaerobisk nedbrytningsprocess. Käppalaförbundet driver det tredje största reningsverket i Sverige. I framtiden förväntas antal anslutna hushåll till reningsverket att öka samt strängare slamhanteringskrav appliceras. Detta innebär att nuvarande slamstrategi måste utvecklas. En idé är att tillämpa en termofilrötnings process. En termofil rötningsprocess har visat sig ha högre kapacitet, lägre uppehållstider och ökad patogen reduktion i jämförelse med en mesofil rötning. En nackdel med termofil rötning är den försämrade processtabiliteten.  I denna studie har en 5 m3 pilotanläggning inhyrd från RISE använts för att utvärdera den maximala organiska belastningen för en stabil termofil rötningsprocess med blandslam som substrat. Fyra uppehållstider valdes och varje uppehållstid hölls tre gånger. Regelbundna analyser av rå och rötslam utfördes för att erhålla information om process stabilitet och effektivitet. Pilotanläggningen styrdes genom ett människa-maskingränssnitt där driftparametrar matades in. Huvudmålen för studien var att se hur en ökning av organisk belastning påverkade pH, alkaliniteten och halt av flyktiga fettsyror samt att undersöka alkalinitetens och halten av flyktiga fettsyrors påverkan på processtabiliteten. Påverkan av en ökad organisk belastning på gas sammansättning, gasproduktion, rötningsgrad har även undersökts. Resultaten visade att flyktiga fettsyror initialt ökade och alkaliniteten minskade vid en belastnings ökning. Flyktiga syror och alkalinitet återgick till mer stabila värden efter processen fick återhämta sig vid en konstant belastning. Belastningsökningen orsakade små variationer i pH värdet men inte tillräckligt för att orsaka processkollaps. En ökad belastning verkade inte ha någon större påverkan på rötningsgraden då den låg runt 40–50% hela rötningsperioden. Skumutvecklingen visade sig inte heller vara ett problem. Resultaten visade även att väte sulfid ökade i gasen när en belastningsökning gjordes. Den volymetriska gas och metan produktionen ökade vid en ökad OLR medan specifika gas och metan produktionen minskade. Slutsatsen var att en stabil termofil rötningsprocess av blandslam kan drivas med en organisk belastning på 6.55 ± 0.06 kg VS m-3d-1 och en uppehållstid på 7 dagar. Maximal belastning för stabil termofil rötningsprocess uppnåddes dock ej då processen förblev relativt stabil under alla undersökta uppehållstider. Processen återhämtade sig även efter tecken på instabilitet vilket indikerar att AD processen kan behålla stabilitet vid de undersökta belastningar.

Les cheminées et sédiments hydrothermaux actifs du Bassin de Guaymas (Mexique) hébergent diverses communautés microbiennes présentant différents métabolismes, y compris ceux impliqués dans les cycles biogéochimiques du soufre et du fer. Il est établi, que, dans ces écosystèmes dynamiques, les micro-organismes qui dépendent pour leur croissance des substrats présents dans leur environnement pourraient à leur tour, affecter localement la composition du fluide hydrothermal et la minéralogie des cheminées, la composition des sédiments, en favorisant la dissolution et/ou la précipitation de certaines phases minérales. Pour étudier ces interactions procaryotes-biotopes, et établir des liens entre la structure, l’activité et les signatures isotopiques des communautés microbiennes et les caractéristiques physico-chimiques des systèmes hydrothermaux du Bassin de Guaymas, une approche pluridisciplinaire mettant en oeuvre des techniques de microbiologie, de géochimie et d’isotopie, lors des cultures d’enrichissements en bioréacteur et des études de colonisation de substrats basaltiques a été réalisée. Ces travaux ont permis : 1) de caractériser la diversité des micro-organismes de différents compartiments de l’écosystème hydrothermal (cheminée et sédiments) du Bassin de Guaymas par des approches culturales et moléculaires ; 2) de décrypter le fonctionnement des cycles biogéochimiques du soufre et du fer en mettant en évidence la présence simultanée, à haute température et en anaérobiose, de micro-organismes sulfo-réducteurs, sulfato-réducteurs, sulfo-oxydants mais également ferri-réducteurs et ferro-oxydants ; et 3) de mieux comprendre les interactions procaryotes-minéraux-fluide en soulignant le rôle des microorganismes dans la modification des conditions environnementales, la formation de minéraux ainsi que leur rôle direct ou indirect dans les fractionnements des isotopes du Fer et Soufre. Ces études ont donc permis de montrer l’importance des interactions croisées entre les différents composantes : communautés microbiennes, le fluide (fluide hydrothermal, eaux interstitielles des sédiments, eau de mer) et les structures minérales des écosystèmes hydrothermaux. / Active hydrothermal chimney and sediments of the Guaymas Basin (Mexico) host various microbial communities with different metabolisms, including those involved in biogeochemical cycles of sulfur and iron. It is established that, in these dynamic ecosystems, microbial activity depends on the availability of substrates in their environment and that prokaryotes could, in return locally affect the composition of the hydrothermal fluid and mineralogical composition of the chimney or sediment, by mediating the dissolution and / or precipitation of some mineral phases. In order to study these prokaryotes-biotope interactions, and establish links between the structure, the activity and the isotopic signatures of microbial communities with the physico-chemical characteristics of hydrothermal systems of Guaymas Basin, a multidisciplinary approach based on the implementation of microbiology, geochemistry and isotope technics was performed using enrichment cultures in bioreactors and the study of the colonization of basaltic substrates. This work led to: 1) characterize the diversity of microorganisms in different hydrothermal compartments (chimney and sediments) of the Guaymas Basin using both cultural and molecular approaches; 2) to decrypt the functioning of sulfur and iron biogeochemical cycles, by highlighting the occurrence, at high temperature and anaerobic condition, of sulfur-reducing, sulfate-reducing, sulfur-oxidizing and also iron-reducing and iron-oxidizing prokaryotes ; and 3) to better understand the prokaryote-mineral-fluid interactions by pointing out the role of microorganisms in environmental condition changes, mineral precipitation and their direct or indirect role in iron and sulfur isotope fractionation. Thus these studies have demonstrated the importance of the cross-interactions between the different hydrothermal components: microbial communities, fluids (hydrothermal fluid, sediment pore water, sea water) and mineral structures.