Thesis (MScEng) -- Stellenbosch University, 2014. / ENGLISH ABSTRACT: Urine contains up to 80% of nitrogen, 50 % of phosphates and 90 % of potassium of the total load in domestic wastewater but makes up less than 1% of the total volume (Larsen et al., 1996). The source separation and separate treatment of this concentrated waste stream can have various downstream advantages on wastewater infrastructure and treated effluent quality. The handling of undiluted source separated urine however poses various challenges from the origin onward. The urine has to be transported to a point of discharge and ultimately has to be treated in order to remove the high loads of organics and nutrients. Wilsenach (2006) proposed onsite treatment of source separated urine in a sequencing batch reactor before discharging it into the sewer system. This study focused on the treatment of urine in a sequencing batch reactor (SBR) primarily for removal of nitrogen through biological nitrification-denitrification. The aim of the study was to determine nitrification and denitrification kinetics of undiluted urine as well as quantification of the stoichiometric reactions. A further objective was to develop a mathematical model for nitrification and denitrification of urine using experimental data from the SBR. The SBR was operated in 24 hour cycles consisting of an anoxic denitrification phase and an aerobic nitrification phase. The sludge age and hydraulic retention time was maintained at 20 days. pH was controlled through influent urine during volume exchanges. Undiluted urine for the study was obtained from a source separation system at an office at the CSIR campus in Stellenbosch. Conditions in the reactor were monitored by online temperature, pH and ORP probes. The OUR of the system was also measured online. One of the main challenges in the biological treatment of undiluted urine was the inhibiting effect thereof on nitrification rate. The anoxic mass fraction was therefore limited to 17 % in order to allow longer aerobic phases and compensate for the slow nitrification rates. Volume exchanges were also limited to 5% of the reactor volume in order to maintain pH within optimal range. Samples from the reactor were analysed for TKN, FSA-N, nitrite-N, nitrate-N and COD. From the analytical results it was concluded that ammonia oxidising organisms and nitrite oxidising organism were inhibited as significant concentrations of ammonia-N and nitrite-N were present in the effluent. It was also concluded that nitrite oxidising organisms were more severely inhibited than ammonia oxidising organisms as nitrate-N was present in very low concentrations in the effluent and in some instances not present at all. Ultimately the experimental system was capable of converting 66% of FSA-N to nitrite- N/nitrate-N of which 44% was converted to nitrogen gas. On average 48% of COD was removed. A mathematical model was developed in spreadsheet form using a time step integration method. The model was calibrated with measured online data from the SBR and evaluated by comparing the output with analytical results. Biomass in the model was devised into three groups, namely heterotrophic organisms, autotrophic ammonia oxidisers (AAO) and autotrophic nitrite oxidisers (ANO). It was found that biomass fractionation into these three groups of 40% heterotrophs, 30% AAO and 30% ANO produced best results. The model was capable of reproducing the general trends of changes in substrate for the various organism groups as well as OUR. The accuracy of the results however varies and nearexact results were not always achievable. The model has some imperfections and limitations but provides a basis for future work.

Master of Science / Department of Civil Engineering / Alok Bhandari / In the upper Midwest, subsurface tile drainage water is a major contributor of nitrate (NO[subscript]3–N) coming from fertilizers and animal manure. Movement of NO[subscript]3-N through tile drainage into streams is a major concern as it can cause eutrophication and hypoxia conditions, as in the Gulf of Mexico. Denitrifying bioreactors is one of the pollution control strategies to treat contaminated tile drainage water. These bioreactors require four conditions which are: 1) organic carbon source, 2) anaerobic conditions, 3) denitrifying bacteria and 4) influent NO[subscript]3-N. This research focuses on investigating fate of veterinary antibiotics in woodchips commonly used in in-situ reactors. Tylosin (TYL) and sulfamethazine (SMZ) are two veterinary antibiotics which are most commonly used in the United States and can be found in tile water after manure is land applied. Partition coefficients of TYL and SMZ on wood were determined by sorption experiments using fresh woodchips and woodchips from an in situ reactor. It was concluded that the woodchips were an effective means to sorb the veterinary antibiotics leached into the tile water after application of animal manure. Linear partition coefficients were calculated and phase distribution relationships were established for both the chemicals. The fresh woodchips gave inconclusive data but predictions could be made by the information determined in the experiments using woodchips from a ten year old woodchip bioreactor. Desorption was also studied and the likelihood of desorption was predicted using the Apparent Hysteresis Index. Overall, it was found that the old woodchips allowed for quick sorption of both antibiotics. It was also found that SMZ had reversible sorption on old woodchips. Thus, it was concluded that the woodchip bioreactor would not be effective for removal of veterinary antibiotics from tile drainage. More research is required for the fate of TYL and to confirm the conclusion.

Dans l'étang de Berre, les apports excessifs en eau douce entraînent souvent des variations brusques de la salinité et des apports en azote parfois abondants. Les processus de régénération sédimentaire et d'élimination d'azote constituent des phénomènes majeurs pouvant expliquer le développement phytoplanctoniques et contrôler l'eutrophisation. L'étude de ces processus a été réalisée dans 2 stations ayant subi des historiques de variations de salinité différents. RDNA et anammox étaient négligeables. La dénitrification (couplée à la nitrification) serait le principal processus de sortie de N2 de l'étang. La forte production primaire pélagique utilisant l'ammonium a pu être expliquée par les processus de régénération, contrairement au nitrate dont la demande pélagique serait soutenue par des apports externes. Comme la salinité est susceptible d'influencer les processus d'azote, des microcosmes ont été utilisés afin d'évaluer la sensibilité de ces processus (résistance, résilience, shift) face aux variations à court terme de la salinité. Les 2 stations ont présenté des réponses différentes (dénitrification et nitrate réduction plus résilientes que la nitrification), suggérant que les stress halins à long terme influencent les réponses à court terme. Une hypothèse serait que la diversité et la structure des communautés joueraient un rôle crucial dans le maintien des taux d'activité. Dans cette étude, seule la communauté nitrifiante (AO) a été étudiée. Les AO et leurs transcrits étaient spécifiques aux sites et leurs abondances changeaient faiblement entre les microcosmes (résistance plus forte dans le site fréquemment soumis à des fluctuations de salinité). / The Berre lagoon receives excess freshwater leading to strong changes in the ecosystem salinity and in the nutrient inputs. Regeneration processes and the N2 removal of nitrogen are of a particular interest as they can explain the planktonic development and control the eutrophication state. These processes were studied in 2 stations with different patterns of long term stresses. DNRA and anammox were negligible. Denitrification (particularly the one coupled to nitrification) would be the main N2 removal in the lagoon. The high primary production based on N-NH4+ can be explained by the regeneration processes, while the primary production based on N-NO3- might be sustained by external inputs. As salinity susceptible to influence directly or indirectly the nitrogen processes, a microcosm approach was used in order to assess their sensitivity (resistance, resilience, shifts) to short term salinity fluctuations. Overall, towards the same kind of perturbations, the two stations responded differently (denitrification and nitrate reduction exhibited showed higher resilience than nitrification.), suggesting that long-term saline stresses would influence short-term responses. We suggest that the diversity and the structure of the communities would be crucial in their functional redundancy and thereby in the maintenance of the rates. In this study, we focused on the total structure of the nitrifying community (AO). AO and their transcripts were site-specific and their abundances changed slightly between microcosms (higher resistance in the site subject to frequent salinity fluctuations compared to the other site which is not affected by changes in the in situ salinity).

Objetivou-se neste trabalho avaliar a influência do tempo de detenção hidráulica (TDH) na remoção do surfactante aniônico, alquilbenzeno linear sulfonado (LAS), de água residuária de lavanderia, em dois reatores de leito fluidificado (RLF) operados em TDH de 8 horas (RLF1) e 12 horas (RLF2). Os reatores foram inoculados com biomassa de lodos ativados e foi investigado a influência da adição de nitrato e etanol na degradação do LAS e seus impactos na comunidade microbiana. Para tal fim, o trabalho foi desenvolvido nas fases que se seguem: caracterização da água residuária de lavanderia comercial, ensaio de degradação desnitrificante de LAS, operação dos reatores de leito fluidificado, planejamento experimental de condições ótimas de degradação de surfactante em reatores em batelada e caracterização microbiana do consórcio microbiano do RLF1 e RLF2 por meio do Illumina MiSeq. Verificou-se considerável remoção de LAS (77%) no ensaio de degradação sob condição desnitrificante nos reatores em batelada com 21 mgLAS/L e adição de 100 mgNO3-/L. Ao longo de 436 dias de operação, verificou-se elevada remoção de LAS em RLF1 e RLF2, todavia, observou-se diferença significativa; ou seja, maior remoção global de LAS (99±3%) para 22,5±5,9 mg /L afluente em RLF2. A adição de etanol influenciou, notadamente, a remoção de LAS em RLF1, por outro lado a adição de nitrato exerceu influência na remoção de LAS em ambos os reatores. Na realização do planejamento experimental, em reatores em batelada, pode-se obter condições ótimas de degradação de LAS, verificando-se os valores críticos para etanol de 97,5 mg /L, e de 87,5 mg /L para nitrato. Para as seis etapas de operação foram identificados gêneros relacionados à desnitrificação e a degradação de LAS. Para Etapa I, foram identificadas bactérias relacionadas com a redução de nitrato, dentre eles Flavobacterium, Sphingobium e Xanthobacter. Para as etapas com adição de surfactante (II, III, IV, V e VI) identificou-se Dechloromonas, Zoogloea, Novosphingobium, Pseudomonas, Acinetobacter e Desulfobulbus, provavelmente relacionadas à degradação de LAS. / The aim of this study was to evaluate the influence of hydraulic retention time (HRT ) on anionic surfactant; linear alkyl benzene sulphonate (LAS) from laundry wastewater in two fluidized bed reactors (FBR) operated in HRT of 8 hours (FBR1) and 12 hours (FBR2). The reactors were inoculated with activated sludge and the influence of nitrate and ethanol addition on LAS removal was studied with subsequent impact on microbial diversity. The study was conducted in following phases; laundry wastewater characterization, denitrifying degradation potential of LAS, fluidized bed reactor operation under specified conditions, central composite design, and determination of microbial shift between biofilm of FBR1 and FBR2 through Mi-Seq Illumima®. Higher LAS removal (77%) was verified in denitrifying potential degrading of LAS using batch reactors feed with 21 mg LAS /L and 150mg NO3-/L. Great LAS removal efficiency was observed for 436 days, however significant notice was observed for global LAS removal between FBR1 and FBR2, i.e. greater LAS removal in FBR2 (99±3% for 22,5 ±5,9 mg LAS/L) than FBR1 (93 ±12% for 20,6 ±4,4 mg LAS /L). The ethanol addition significantly influenced FBR1 efficiency while nitrate addition improves LAS removal in both reactors. Central composite design resulted into optimal values for LAS removal (94%) in addition to 97,5 mg/L ethanol and 87,5 mg/L nitrate. Microbial shift showed genus abundance related to denitrification and LAS degradation. In Step I denitrifying bacteria Flavobacterium, Shingobium and Xanthobacter were identified. For Steps with laundry wastewater addition, Dechloromonas, Zoogloea, Novosphingobium, Pseudomonas, Acinetobacter and Desulfobulbus, were identified, probably, related to LAS degradation.

O nitrogênio é o quinto elemento mais abundante em nosso sistema solar, é essencial para a síntese de ácidos nucleicos e proteínas, os dois polímeros mais importantes da vida. Apesar da importância do nitrogênio e sua abundância na atmosfera, o N2 é praticamente inerte; assim, as formas de nitrogênio inorgânico fixados mais comuns são os íons nitrato e amônio que muitas vezes limitam a produtividade primária nos ecossistemas marinhos e terrestres. Uma biosfera ativa requer a incorporação do nitrogênio em moléculas biológicas por meio da fixação de nitrogênio, um processo no qual os domínios em procarióticos Bacteria e Archaea reduzem gás nitrogênio em amônia. Porém, a maioria dos organismos não pode fixar nitrogênio, mas sim obtê-lo na forma de nitrogênio orgânico a partir do ambiente, ou a partir da redução do nitrato. O amônio é retornado para o ambiente quando os organismos morrem e isto depende da presença do oxigênio nestes locais. Na presença do oxigênio, o amônio é oxidado em nitrato, numa via conhecida como nitrificação. Na ausência de oxigênio, o nitrato é utilizado por muitos microrganismos como um receptor de elétrons, como na redução dissimilatória de nitrato a amônia (DNRA), acoplada à oxidação anaeróbica de carbono orgânico; o nitrato pode ser convertido em gás dinitrogênio pela desnitrificação, ou ainda uma via alternativa pode transformar o nitrogênio fixado em N2, realizada por um grupo de bactérias conhecido como planctomycetes, onde a oxidação do amônio é acoplada a redução de nitrato, no processo chamado anammox. Neste trabalho de tese foi realizada a avaliação do ciclo do nitrogênio, buscando entendimento de três vias de transformação deste elemento. Neste sentido, estas vias foram avaliadas quanto as suas taxas de ocorrência, com uso incubações do sedimento de manguezais e nitrogênio marcado, como também por meio da identificação de genes funcionais envolvidos nestes três processos, por meio de pirosequenciamento. O grupo Planctomycetes ainda pode ser estudado utilizando a hibridização com sonda fluorescente in situ FISH. Os resultados indicam a presença marcante de genes identificados como parte da via desnitrificação, assim como as taxas de incubação observadas para esta via foram muito expressivas quando comparadas com as taxas obtidas para o processo anammox e DNRA. Os resultados obtidos com as análises metagenômicas e com a técnica FISH sustentam os resultados obtidos com as incubações. Pode-se observar que a filogenia sugere a tendência do agrupamento com organismos planctomycetes não descritos como responsáveis pelo processo anammox. Por meio da técnica FISH pode-se confirmar a detecção destas bactérias e visualizar sua menor presença nos manguezais amostrados, quando comparadas às bactérias totais encontradas nestas amostras. / Nitrogen is the fifth most abundant element in our solar system, essential for the synthesis of nucleic acids and proteins, one of the most important polymers of life. Despite the importance of nitrogen and its overwhelming abundance in the atmosphere, N2 is practically inert, so the most commom forms of inorganic nitrogen fixed are nitrate and ammonium ions, which often limit primary productivity in marine and terrestrial ecosystems. An active biosphere requires the incorporation of nitrogen in biological molecules through nitrogen fixation, a process in which domains Archaea and Bacteria reduce nitrogen gas into ammonia. The most organisms can\'t fix nitrogen, but they can get it in inorganic nitrogen form from the environment or from the reduction of nitrate. The ammonia is returned to environment when organisms die and it depends on the presence of oxygen at these sites. In the presence of oxygen, ammonia is oxidized to nitrate, in a way known as nitrification, and in the absence of oxygen, the nitrate being used by many microbes as an electron acceptor, such as dissimilatory nitrate reduction to ammonia (DNRA), coupled to anaerobic oxidation of organic carbon. The nitrate can be converted to dinitrogen gas by denitrification, or an alternative route can turn fixed nitrogen in N2, performed by a bacteria group known as Planctomycetes, where oxidation is coupled to the ammonium nitrate reduction, in a process called anammox. In this work, with mangrove sediments from Bertioga city, State of São Paulo, the evaluation of these three pathways of nitrogen transformation was performed as their rates of occurrence, with incubations with labeled nitrogen, as well as through the identification of functional genes involved in these three cases, by pyrosequencing. The group Planctomycetes can also be studied using the hybridization probe FISH fluorescence in situ. The results show the strong presence of genes identified as part of the denitrification pathway, as well as the rates observed for incubation of this pathway were very significant when compared with the rates obtained for the process anammox and DNRA. The results obtained from the metagenomic analyze and the FISH technique support the results obtained with incubations. It can be observed that the trend suggests the phylogeny of the organisms group with Planctomycetes not described as responsible for anammox process. Through the FISH technique, it was possible to confirm the detection of these bacteria and view its reduced presence in mangrove sampled compared to total bacteria found in these samples.

A presente pesquisa teve a intenção de desenvolver reator biológico de 159 \'M POT.3\' de volume total, com leito de carvões granulares (ativado e antracito) expandido e parcialmente aerado, para promover remoções de matéria orgânica e nutrientes (N e P) do esgoto sanitário, devido a manutenção de ambientes anaeróbio e aeróbio, estratificados ao longo da altura do reator e ao longo do biofilme cultivado em seu interior. Para avaliar o impacto das modificações operacionais no comportamento biológico dos microrganismos presentes no reator, foram aplicados métodos para análise das atividades nitrificante (ANE), desnitrificante (ADE) e metanogênica (PME), embasados em testes respirométricos padrões descritos na literatura. A estratificação reacional também foi estudada dentro de filmes biológicos com a ajuda de microsensores amperométricos de OD em testes de bancada. Assim, após 451 dias de operação, foi possível verificar remoções médias de \'DQO IND.F\', NTK e fosfato total de, respectivamente 78%, 56% e 42%, quando o reator foi operado por 66 dias com tempo de detenção hidráulica médio de 8,8 h, injetor de oxigênio puro, instalado após placa de orifício em linha de recirculação aerada, a qual funcionava pressurizada (3 a 4 bar) e com razão média de recirculação igual a 3, comparada à vazão de alimentação. Para o restante do período, no qual foram aplicadas taxas de carregamentos volumétricos médios de 0,74 \'+ OU -\' 0,28 kg\'DQO IND.F\'/\'M POT.3\'.dia; 0,17 \'+ OU -\' 0,07 kg NTK/\'M POT.3\'.dia e 0,05 \'+ OU -\' 0,02 kg\'PO IND.4\'POT.-3\'/\'M POT.3\'.dia, o sistema demonstrou remoções médias de 65 \'+ OU -\' 20% para \'DQO IND.F\', 25 \'+ OU -\' 21% para o NTK e 48 \'+ OU -\' 18% para o fosfato total, mesmo operado com tempo de retenção celular médio de 15 \'+ OU -\' 7 dias. Os testes respirométricos foram sensíveis para avaliar a atividade microbiana do material biológico coletado ao longo do reator, com os quais foram verificados PME médio de 0,25 mL\'CH IND.4\'/gSVT.h, para as amostras de material biológico coletado na região anaeróbia, ao fundo do reator; ANE variando entre 1,3 a 4,4 mg\'O IND.2\'/gSVT.h, para as amostras coletadas na região aeróbia, intermediária ao reator; e ADE variando entre 0,024 e 5,20 mg\'N IND.2\'/gSVT.h, para amostras coletadas, respectivamente, no fundo e no topo do reator. As análises do material líquido também corroboram com a idéia de estratificação dos ambientes aeróbio, no fundo do reator, e micro-aerado, em sua região intermediária superior, apontando para região com alto potencial de ocorrerem nitrificação e desnitrificação conjuntas no topo da sua zona reativa. O microsensor amperométrico de OD com ponta entre 10 e 30 \'mü\'m de diâmetro, confeccionados em laboratório especializado, se mostrou sensível para a observação do gradiente de concentração de oxigênio dissolvido dentro de filme biológico, suficientes para a formação de regiões aeróbia e anaeróbia em seu interior, cuja informação pode ser útil para a aprimoramento de reatores com biofilme, projetados para remoções combinadas de matéria orgânica e nutrientes. Entretanto, quando se pretende definir parâmetros cinéticos ou de transferência de massa, maior rigor deve ser dispensado na definição dos locais para a aplicação do sensor, nos quais sejam reduzidos os efeitos da heterogeneidade do agregado microbiano no ajuste da modelagem matemática aplicada aos pontos experimentais / The current research aimed the development of a 159 \'M POT.3\' total volume biological reactor, with a expanded and partially aerated granular coal bed (activated and anthracite), to promote organic matter and nutrients (N and P) removal from wastewater due to the maintenance of anaerobic and aerobic environments, stratified throughout the height of the reactor and also all over the biofilm cultivated in its interior. Methods for the analyses of specific nitrifying activities (ENA), denitrifying (EDA) and methanogenic (EMA) were applied to assess the impact of the operational modifications in the biological behavior of microorganisms present in the reactor, based on standard respirometric tests found in literature. The reactional stratification was also studied inside the biological films with the help of DO microsensors in batch tests. Thus, after 451 of operation it was possible to verify mean \'COD IND.F\', TNK and phosphate removal of 78%, 56% and 42%, respectively, when the reactor was operated for 66 days with average hydraulic detention time of 8.8 h, pure oxygen injector which was installed after the aerated recirculation line which was working pressurized (3 to 4 bar) and with mean recirculation ratio equal to 3, when compared to the feeding flow. For the rest of the period where mean volumetric loading rates of 0.74 \'+ OR -\' 0.28 kg\'COD IND.F\'/\'M POT.3\'.day; 0.17 \'+ OR -\' 0.07 kgTNK/\'M POT.3\'.day and 0.05 \'+ OR -\' 0.02 kg\'PO IND.4\'POT.-3\'/\'M POT.3\'.day were applied the system demonstrated average removal of 65 \'+ OR -\' 20% for \'COD IND.F\', 25 \'+ OR -\' 21% for TNK and 48 \'+ OR -\' 18% for total phosphate, even when it operated with mean cellular retention time of 15 \'+ OR -\' 7 days. The respirometric tests were sensible enough to assess the microbial activity from the biological material collected throughout the reactor, and where mean PME of 0.25 mL\'CH IND.4\'/gSVT.h was verified for the samples of biological material collected in the anaerobic region, at the bottom of the reactor; ANE varying between 1.3 to 4.4 mg\'O IND.2\'/gSVT.h, for the samples collected in the aerobic region, reactor\'s intermediary; and ADE varying between 0.024 to 5.20 mg\'N IND.2\'/gSVT.h, for samples collected at the bottom and the top of the reactor, respectively. The analyses of the liquid material also support the idea of stratification of the aerobic environments, at the bottom of the reactor, and micro-aerated, in its superior intermediate region, pointing to the region as a high potential of occurring joint nitrification and denitrification at the top of the reactive zone. The DO amperometric microsensor, with tip between 10 and 30 \'mü\'m of diameter, produced in a special laboratory, is sensible to the determination of dissolved oxygen concentration gradient inside biological film, sufficient for the formation of anaerobic and aerobic regions in its interior, this information can be useful to the improvement of biofilm reactors, projected for the combined removal of organic matter and nutrients. However, when the definition of kinetic parameters or mass transference is intended more strictness must be applied when choosing the locals for microsensor application, where the effects of the microbial aggregate heterogeneity is reduced in the adjustment of the mathematical modeling applied to the experimental points.

A ampla ocorrência de contaminação de águas por compostos de nitrogênio em concentrações superiores às recomendadas pela legislação tem suscitado interesse no desenvolvimento de tecnologias viáveis de remoção desses compostos. A remoção biológica de nitrogênio apresenta como principais vantagens os custos relativamente reduzidos e a possibilidade de maior eficiência. Compostos reduzidos de enxofre como sulfetos podem ser oxidados a enxofre elementar ou a sulfato por bactérias oxidantes de sulfeto que utilizam nitrato ou nitrito como receptor de elétrons. Esta desnitrificação reduz os requerimentos globais de carbono para a remoção de nutrientes, com menor produção de lodo, proporcionando grande economia. O objetivo desta pesquisa consistiu em contribuir para o conhecimento acerca dos aspectos microbiológicos do processo de desnitrificação com o uso de sulfeto. Foram avaliados os efeitos dos modos de operação dos reatores desnitrificantes sobre a biomassa em cada uma das diferentes configurações e monitorada a colonização microbiana por meio de técnicas de Biologia Molecular como PCR/DGGE, sequenciamento e análises filogenéticas. Os fragmentos do gene RNAr 16S foram relacionados aos gêneros Pseudomonas, Aeromonas, Acidobacteria, Chlroroflexi, Clostridium, Cupriavidus e Ralstonia. Filotipos dos clones para o sistema piloto foram associados a bactérias não cultiváveis e Firmicutes envolvidos na digestão anaeróbia em reatores tratando água residuária, Synergistetes, Deferribacteria e Proteobacteria. Foram identificados micro-organismos presentes nos reatores com reconhecida capacidade para desnitrificação: Pseudomonas, Desulfovibrio desulfuricans e Ralstonia. Amostras de ambos os reatores desnitrificantes apresentaram reações de amplificação positivas com primers específicos para bactérias semelhantes a Thiomicrospira associados a primers universais: 100% das amostras amplificaram com OST1F/1492R e 75% com EUB8F/OSTR1R. Para duas condições de operação do reator em escala de bancada foram identificados micro-organismos semelhantes a Sulfurimonas denitrificans, bactéria autotrófica redutora de nitrato e oxidadora de sulfeto. Chloroflexi também foram encontrados em digestores localizados em plantas de tratamento de águas residuárias recebendo essencialmente efluente doméstico e Propionibacterium foi associada a comunidade microbiana de reatores UASB tratando água residuária industrial. Bactérias em associações sintróficas com participação no ciclo do enxofre e/ou na digestão anaeróbia foram igualmente identificadas: Clostridium sulfidigenes e outros Firmicutes, Synergistetes, clones de bactérias de cultura de enriquecimento e bactérias do gênero Syntrophorhabdus. Os resultados deste trabalho proporcionaram associar a colonização microbiana com o desempenho e as características metabólicas de alguns micro-organismos com reatores desnitrificantes combinados para o tratamento de águas residuárias. / The widespread nitrate contamination in concentrations higher than recommended by legislation has raised interest in technologies for water and wastewater treatment. Biological nitrogen removal is relatively low cost and higher efficiency. Sulfide as electron donor can be oxidized to elemental sulfur or sulfate by sulfide oxidizing bacteria that can use nitrate or nitrite as electron acceptor. This type of denitrification reduces the overall requirements for removal of carbon nutrients and less sludge is produced. The aim of this research was to contribute to microbiological knowledge about denitrification using sulfide. The effects of operation conditions on the denitrifying biomass were monitored through molecular biology techniques such as PCR/DGGE, sequencing and phylogenetic analysis. 16S rRNA gene fragments were related to Pseudomonas, Aeromonas, Acidobacteria, Chlroroflexi, Clostridium, Cupriavidus and Ralstonia. Phylotypes of clones from samples of pilot-scale reactor were associated with non-cultivable bacteria and Firmicutes involved in anaerobic digestion of wastewater, Synergistetes, Deferribacteria and Proteobacteria. Microorganisms with ability to denitrification were identified in both reactors: Pseudomonas, Desulfovibrio desulfuricans and Ralstonia. Samples of denitrifying reactors showed positive amplification with specific primers for Thiomicrospira associated to universal primers: 100% of the samples amplified with OST1F/1492R and 75% EUB8F/OSTR1R. Sulfurimonas denitrificans-like were identified for two operational conditions of the bench scale reactor. Chloroflexi were also found in treatment plants digesters receiving domestic wastewater and Propionibacterium was associated with microbial community of UASB reactors treating industrial wastewater. Syntrophic bacteria participating in the sulfur cycle and/or anaerobic digestion were also identified: Clostridium sulfidigenes and other Firmicutes, Synergistetes, clone enrichment culture bacteria and Syntrophorhabdus. These results provide to associate this microbial colonization and metabolic characteristics of some microorganisms with performance of combined denitrifying reactors for treatment of wastewater.

Um Reator com Aeração Intermitente e Leito Estruturado (RAILE) foi testado como pós- tratamento de UASB tratando efluente doméstico. O RAILE foi submetido a ciclos de uma hora de aeração e duas de anoxia, visando obtenção de Nitrificação e Desnitrificação Simultâneas (NDS) para remoção de nitrogênio. A recirculação aplicada foi de 3 vezes a vazão de entrada, para equalização da qualidade do efluente. O reator tem formato cilíndrico e seu meio suporte é espuma de poliuretano na forma de cilindros dispostos verticalmente, suportados por hastes de cloreto de polivinila (PVC) presas nas extremidades. O estudo foi dividido em duas fases em que o RAILE recebeu efluentes de UASB com diferentes relações de DQO/N-Total. Na primeira fase a relação foi 2,44 e na segunda fase a relação foi 4,01. As remoções de N-Total foram 23,7% e 65,3%, respectivamente para primeira e segunda fase. As remoçôes de DQO para todo o sistema foram 94,8%, para a primeira fase, e 93,4%, para a segunda fase. A alcalinidade se mostrou um parâmetro efetivo pra monitorar a ocorrência de NDS. Embora tenha se mostrado viável como pós-tratamento de UASB em sistemas tratando efluente doméstico, a relação DQO/N-Total precisa ser controlada para garantir o processo de NDS / A Structured Bed Intermittently Aerated reactor (SBIA) was tested as post-treatment for UASB treating domestic wastewater. The SBIA underwent cycles of one hour of aeration and two hour of anoxia to obtain Simultaneous Nitrification and Denitrification (SND) for the removal of nitrogen. The recirculation flow applied was 3 times the input flow to equalize the effluent quality. The reactor had a cylindrical shape and its support medium is polyurethane foam in the form of cylinders arranged vertically, supported by polyvinyl chloride (PVC) rods attached at the ends. The study was divided into two stages in which the SBIA received the UASB reactor effluent with different COD/Total-N ratios. The first COD/Total-N ratio was 2,44 and the second was 4,01. The removal of N-total were 23.7% and 65.3% respectively for the first and second stages. The removal of COD, in all the system, was 94.8% for the first stage and 93.4% for the second stage. The alkalinity proved to be an effective parameter to monitor the occurrence of SND. Although the SBIA proved to be feasible as post-treatment of UASB systems treating domestic wastewater the ratio COD/N-total must be controlled to ensure the process of SND

O reator em batelada seqüencial com biomassa em suspensão foi submetido a concentrações de N-amoniacal de 125, 250 e 500 mg N/L e em condições de oxigênio dissolvido (OD) no reator de 2 mg 'O IND.2'/L, em ciclos de 2h/2h de anóxico/aeróbio. Em todas as fases, o reator foi operado como batelada alimentada. Na condição de 125 mg N/L obteve-se eficiência de remoção de 87% de N, tendo o nitrato sido o principal produto da nitrificação. Na condição de 250 mg N/L, obteve-se eficiência de remoção de N de 84%, com predominância de nitrito como principal produto da nitrificação e com ocorrência de nitrificação e desnitrificação simultânea durante os dois primeiros ciclos aeróbios. Na condição de 500 mg N/L, as condições de concentração de OD de 2 mg 'O IND.2'/L e aeração intermitente a cada 2h não foram suficientes para promover a remoção total de nitrogênio amoniacal. Foram feitas alterações: ciclos de 2h anóxico e 9h aeróbios, com concentração média de 2,8 mg 'O IND.2'/L, que resultaram em eficiências de remoção de N de 94%, com predominância de nitrito. Foram isoladas cepas desnitrificantes com similaridade de 97% para Thauera mechernichensis e Thauera sp. 27 nas condições operacionais de 125 e 250 mg N/L e de 99% para Ochrobactrum anthropi e Ochrobactrum tritici, na condição operacional de 500 mg N/L. O longo tempo de operação resultou na diminuição da população de bactérias oxidantes de nitrito, podendo ter sido uma das causas que contribuiu para que se criassem condições que levariam à nitrificação via nitrito na concentração de 500 mg N/L. O sucesso na prevenção da inibição do processo por amônia livre foi atribuído à adoção das condições operacionais do reator, que foi operado sob aeração intermitente e batelada alimentada. / The sequential batch reactor with suspended bioma was subjected to ammonium concentrations of 125, 250 and 500 mg N/L, oxygen dissolved (OD) concentrations of 2 mg 'O IND.2'/L, 2h/2h of anoxic/aerobic steps. The reactor was operated under fed-batch feeding. At the operational condition of 125 mg N/L, the mean nitrogen removal efficiency of 87% was obtained and nitrate was the main nitrification product. At operational condition of 250 mg N/L, was gotten nitrogen removal efficiencies of 84% and nitrite was the predominant form of nitrogen and simultaneous nitrification and denitrification occurred during the two first aerobic steps of the cycle. At 500 mg N/L, the operating conditions imposed to the reactor (OD concentrations of 2 mg 'O IND.2'/L and intermittent aeration of 2h) did not lead to complete nitrification during the aerated steps, thus affecting nitrogen removal. The conditions were altered increasing the aerated steps to 9 hours, with OD concentration of 2,8 mg 'O IND.2'/L, and keeping the duration of the anoxic steps in 2 hours. Under such conditions, the mean nitrogen removal efficiency attained 94% and nitrite was the predominant oxidized nitrogen specie. It was isolated denitrifiers with similarity of 97% for Thauera mechernichensis and Thauera sp. 27 at the operational conditions of 125 and 250 mg N/L and of 99% for Ochrobactrum anthropi and Ochrobactrum tritici, at operational condition of 500 mg N/L. The long term operation resulted in the decrease of nitrite oxidizers populations and this was probably the main factor contributing for the creation of conditions for the partial nitrification via nitrite to prevail during the operation at ammonium concentration of 500 mg N/L. The success in preventing free ammonia inhibition was attributed to the adoption of intermittent aeration and fed batch operation.

Grande parte das indústrias alimentícias, principalmente as que utilizam processos fermentativos, geram efluentes com altas concentrações de material orgânico e nutrientes (principalmente de nitrogênio), necessitando de sistemas complexos para o seu tratamento. Neste sentido, o presente trabalho foi proposto com o intuito de desenvolver um sistema compacto que possa operar como única unidade de tratamento de águas residuárias industriais, tanto na remoção de matéria orgânica quanto na remoção de nitrogênio, com baixo consumo de energia e baixa produção de lodo. Constatou-se, portanto, a viabilidade técnica do reator combinado anaeróbio-aeróbio vertical de leito fixo para o tratamento de efluentes industriais contendo matéria orgânica (1400 mg DQO/l) e nitrogênio (160 mg N/l). A melhor condição operacional do reator foi conseguida aplicando-se tempo de detenção hidráulica (TDH) de 35 h (21 h na zona anaeróbia e 14 h na zona aeróbia), com base no volume útil do reator, e razão de recirculação (R) igual a 3,5, apresentado eficiências na remoção de DQO, NTK e NT de 97%, 94% e 77%, respectivamente, com concentrações efluentes médias de 36 '+ OU -' 10 mg DQO/l, 2 '+ OU -' 1 mg N-'NH IND.4' POT.+'/l,8 '+ OU -' 3 mg N-org/l, 1 '+ OU -' 1 mg N-'NO IND.2'POT.-'/l e 26 '+ OU -' 23 mg N-'NO IND.3'POT.-'/l. O reator vertical de leito fixo, operando unicamente em condição anaeróbia, apresentou eficiências médias na remoção de DQO de 43 '+ OU -' 9%, 60 '+ OU -' 9% e 70 '+ OU -' 6%, respectivamente para TDH aplicados de 11 h, 17 h e 21 h / Most of the food industries, mainly the ones that use fermentative processes, generates effluent with high concentrations of organic matter and nutrients (nitrogen mainly), needing of complex systems for its treatment. In this direction, the present work was considered to develop a compact system that can operate as only unit of industrial wastewater treatment, in the removal of organic matter and nitrogen, with low consumption of energy and low sludge production. Therefore, the viability of the upflow anaerobic-aerobic combined fixed bed reactor for the treatment of industrial effluent with organic matter (1400 mg COD/l) and nitrogen (160 mg N/l) was evidenced. The best operational condition of the reactor was obtained applying 35 h of hydraulic retention time (21 h in the anaerobic zone and 14 h in the aerobic zone) and recycle ratio (R) of 3.5. In this condition, the COD, TKN and TN efficiencies removal were of 97%, 94% and 77%, respectively, with average effluent concentrations of 10 '+ OU -' 36 mg COD/l, 2 '+ OU -' 1 mg N-'NH IND.4'POT.+'/l, 8 '+ OU -' 3 mg N-org/l, 1 '+ OU -' 1 mg N-'NO IND.2'POT.-'/l and 26 '+ OU -' 23 mg N-'NO IND.3'POT.-'/l. The up-flow fixed bed reactor, operating only in anaerobic condition, presented average efficiencies in the COD removal of 43 '+ OU -' 9%, 60 '+ OU -' 9% and 70 '+ OU -' 6%, for HRT applied of 11 h, 17 h and 21 h, respectively