Made available in DSpace on 2017-07-10T19:23:58Z (GMT). No. of bitstreams: 1 Izabela Regina Costa _Araujo.pdf: 2798947 bytes, checksum: 10a0bad3f765462b54bb964dd971203c (MD5) Previous issue date: 2015-02-19 / Researches have shown that among several ways of treating cassava starch wastewater, anaerobic process has been considered a simple and economical option that combines the high content of organic matter conversion to biogas production. On the other hand, biogas is a potential energy resource that can be used to supply part of the energy demands from industrial plants. Thus, this trial aimed at evaluating an anaerobic reactor with fixed bed up-flow, regarding the removal of organic load in wastewater from cassava starch extraction as well as biogas production. So, volumetric organic loads (VOC) of 2.5; 5.0; 8.0 and 10.0 g.L-1.d-1 were tested in a 2.82 L volume reactor. A support medium of polypropylene rings filled with polyurethane foam was used. The monitored parameters regarding the reactor effluent were: pH, volatile acidity (VA), total alkalinity (TA), VA/TA ratio, total solid concentrations (TS), volatile solids (VS), total suspended solids (TSS), volatile suspended solids (VSS), total chemical oxygen demand (TCOD), soluble chemical oxygen demand (SCOD) and organic acids. The reactor performance was tested in relation to removals of solids, organic load, as well as biogas and methane production. As the most important results, it was observed that the reactor remained constant concerning pH, total removal of VSS and removals of TCOD and SCOD. While, there were also lactic, butyric, propionic and acetic acids in the studied effluent, although acetic acid was recorded in higher concentrations. Biogas production average was 3.8 L.d-1, the average of specific biogas production was 0.156 L.gCODconsumed-1, but VOC answer was 10.0 g.L-1.d-1, being the greatest results for both of them. Methane percentage in biogas ranged from 66 to 84% and the specific methane production ranged from 0.01 to 0.36 Lg-1. The highest values were obtained in VOC - 2.5 and 10.0 g.L-1d-1, respectively. The removal of medium TS, VS, TSS and VSS were 72, 80, 63 and 92%, respectively. The average removals of TCOD and SCOD were 98.7 and 97.9 respectively, therefore, the highest removal rates were obtained for 10.0 and 2.5 VOC g.L-1.d-1. Thus, it can be concluded from this trial that it is possible to produce biogas with a methane percentage higher than 80% (maximum) and remove the organic loading rate by more than 90% (maximum) according to tested configuration systems with wastewater from cassava starch industry. Furthermore, the highest biogas productions were obtained for the highest VOC, while it was observed a bias on biogas production with VOC increase as well as the specific production of methane. / Dentre as diversas formas existentes de tratar os efluentes da indústria de fécula de mandioca, o processo anaeróbio é considerado uma alternativa simples e econômica, que alia o elevado grau de conversão da matéria orgânica à produção do biogás. O biogás, por sua vez, é um recurso energético potencial que pode ser utilizado para suprir parte das demandas energéticas das unidades industriais. Nesse sentido, estabeleceu-se como objetivo deste trabalho avaliar um reator anaeróbio de leito fixo e fluxo ascendente, em relação à remoção de carga orgânica da água residuária de extração de fécula de mandioca e à produção de biogás. Para isso, foram testadas as cargas orgânicas volumétricas (COV) de 2,5, 5,0, 8,0 e 10,0 g.L -1.d-1 em um reator de 2,82 L de volume útil. Utilizou-se meio suporte de anéis de polipropileno preenchidos com espuma de poliuretano. Os parâmetros monitorados no efluente do reator foram o pH, acidez volátil (AV), alcalinidade total (AT), razão AV/AT, concentrações de sólidos totais (ST), sólidos voláteis (SV), sólidos suspensos totais (SST), sólidos suspensos voláteis (SSV), demanda química por oxigênio total (DQOT) e demanda química por oxigênio solúvel (DQOS) e de ácidos orgânicos. A eficiência do reator foi avaliada em relação às remoções de sólidos e carga orgânica e à produção de biogás e de metano. Como principais resultados, observou-se que o reator manteve-se estável em relação aos parâmetros pH, remoção de SSV e remoções de DQOT e DQOS. Observou-se a presença de ácido lático, butírico, propiônico e acético no efluente, sendo o ácido acético em maiores concentrações. A produção média de biogás foi de 3,8 L.d-1 e a média da produção específica de biogás foi de 0,156 L.g-1DQOconsumida, sendo que, para ambos os maiores resultados foram obtidos para a COV 10,0 g.L-1.d-1. O percentual de metano no biogás variou de 66 a 84% e a produção específica de metano de 0,01 até 0,36 L.g-1 e os maiores valores foram obtidos nas COV 2,5 e 10,0 g.L-1.d-1, respectivamente. As remoções médias de ST, SV, SST e SSV foram de 72, 80, 63 e 92 %, respectivamente. As remoções médias de DQOT e DQOS foram de 98,7 e 97,9, respectivamente e as taxas de remoção mais altas foram obtidos para as COV 10,0 e 2,5 g.L-1.d-1 . Desta forma, conclui-se que é possível produzir biogás com percentuais de metano superiores a 80% (máximo) e remover a carga orgânica em mais de 90% (máximo), em sistemas com a configuração testada, utilizando água residuária de indústria de fécula de mandioca. Além disso, verificou-se que as maiores produções de biogás foram obtidas para as COV maiores e a tendência de aumento da produção de biogás com o aumento da COV, assim como a produção específica de metano.

In Brazil, with the continued expansion of agriculture for supplying demands from international markets, progressive increases in emissions of green house gases are expected. The purpose of the project was hypothesized with three major approaches, 1) Strategies to mitigate methane emission in small ruminant production systems; 2) Identify tropical plants and individual bioactive compound against methanogenic propertie and 3) In vivo evaluation of the nutrients metabolism of Santa Ines sheep fed with tropical plants. For this, we have performed three experiments. The first study (Expt. 1) was designed to determine the in vitro effects of three tropical tannin rich plants such as Leucaena leucocephala (LL), Mimosa caesalpineafolia (MC), Schinus molle (SM) and one non-tannin rich plant Medicavo sativa (MS) for their anti-methanogenic properties when used with and without polyethylene glycol (PEG). All plants had significantly (P<0.05) influenced the degraded organic matter (TDOM) and degraded neutral detergent fiber (DNDF), especially LL, which had most influence on these parameters compared to other tannin containing plants. LL had positive response on antimethanogenic effects; its nutrient degradability was higher than that of other tannin containing plants. The second study (Expt. 2) was set to evaluate the effect of different organic extracts from the whole plant methanolic extract (MHE) of LL on in vitro gas production and to characterize the chemical constituents by using gas chromatography coupled with mass spectroscopy (GC-MS). Major abundant compounds present at the relative percentages of MHE were found to be stigmasterol trimethyl ester (TMS), neophytadiene, palmitic acid TMS, ?-Linolenic acid TMS and 2, 3, 5, 6-Tetra-M-Anisylbenzene. The effects of additions of different extracts in terms of nutrient degradability (TDOM and DNDF) were increased by all extracts. This study explained that the hexane extract from whole plant MHE was effective against methanogenic activity. The objective of the third study (Expt. 3) was to study the effect of LL plant leaves on rumen fermentation, apparent nutrient digestibility, nitrogen balance and methane production in Santa Ines sheep. The animals were divided in three groups in which they were fed with (i) 88% Tifton 85-hay (Cynodon dactylon) and 12% soyabean meal (Control group, n=4); (ii) 28% Tifton 85-hay (Cynodon dactylon) and 72% LL plus 20 ml solution containing 10g/day/animal of PEG (With PEG group - WPEG, n=6); (iii) 28% Tifton 85-hay (Cynodon dactylon) and 72% LL plus 20 ml of distilled water (Without PEG group- WOPEG, n=6). Nutrient intake (dry matter, organic matter, acid detergent fiber, lignin and crude protein) were higher in WPEG and WOPEG compared to the control group, except neutral detergent fiber intake. Apparent digestibilities and nitrogen metabolism had non-significant effects between the treatments. However, CH4 emissions were significantly lower in WPEG and WOPEG than the control. Furthermore, expressions of microbial populations of methanogens in WPEG had lower tendency than that of WOPEG and control. The most salient findings of this study were that, using plant rich in tannin in diets of small ruminants, we can get more benefits in terms of replacing the source of protein in the diet (food safety) and reduced production of enteric CH4 (animal production) / No Brasil, com expansão da agricultura para suprir as exigências dos mercados internacionais, são esperados aumentos progressivos nas emissões de gases do efeito estufa. O objetivo do projeto foi hipotetisado com três abordagens principais, 1) estratégias para mitigar emissões de metano em sistemas de produção de pequenos ruminantes; 2) identificar plantas tropicais com compostos bioativos com propriedades anti metanogénicas e 3) avaliação in vivo do metabolismo de nutrientes em ovelhas Santa Inês alimentadas com planta taninífera. Para isso, foram efetuados três experimentos. O primeiro estudo (Expt 1) foi concebido para estudar os efeitos in vitro de plantas tropicais ricas em tanino como Leucaena leucocephala (LL), Mimosa caesalpineafolia (MC) e Schinus molle (SM) e uma planta não taninífera, Medicavo sativa (MS) quanto às propriedades anti-metanogénicas quando usadas com e sem polietileno glicol (PEG). Todas as plantas significativamente (P < 0.05) influenciaram na matéria orgânica degradada (MOD) e na fibra em detergente neutro degradada (FDND), especialmente a LL, que teve maior influência sobre estes parâmetros, em comparação com as outras plantas que contém tanino LL teve resposta positiva sobre os efeitos de antimethanogênicos e a degradabilidade dos nutrientes foi maior do que a das outras plantas que contém tanino. O segundo estudo (Expt 2) foi definido para avaliar o efeito de diferentes extratos orgânicos a partir do extrato metanólico da planta (EMP) de LL na produção de gás in vitro e caracterizar os constituintes químicos usando cromatografia gasosa acoplada com espectroscopia de massa (GC-MS). Os compostos mais abundantemente encontrados, em termos de percentagens relativas do EMP, foram o éster de trimetil estigmasterol, neofitadina, ácido palmítico, ácido ?-linolênico e 2, 3, 5, 6-Tetra-M-anizil -benzeno. Os efeitos de adições dos diferentes extratos orgânicos, em termos de degradabilidade de nutriente (MOD e NDFD) foram aumentados para todos os extratos. Este estudo explicou que o extrato de hexano a partir do EMP foi eficaz na atividade anti metanogênicas em modificar a degradação ruminal de nutrientes. O objetivo do terceiro estudo (Expt 3) foi estudar o efeito das folhas da planta LL na fermentação ruminal, digestibilidade aparente de nutrientes, balanço de nitrogênio e produção de metano em ovinos Santa Inês. Os animais foram divididos em três grupos em que eles foram alimentados com (i) 88% feno de Tifton-85 (Cynodon dactylon) e 12% de farelo de soja (Grupo controle, n = 4); (ii) 28% feno de Tifton-85 (Cynodon dactylon) e 72% LL mais 20 ml solução contendo 10g/dia/animal de PEG (grupo com PEG - CPEG, n = 6); (iii) 28% feno de Tifton-85 (Cynodon dactylon) e 72% LL mais 20 ml de água destilada (sem PEG-grupo-SPEG, n = 6). A ingestão de nutrientes (matéria seca, matéria orgânica, fibra em detergente ácido, lignina e proteína bruta) foram maiores no grupos CPEG e SPEG em relação ao grupo controle, exceto a ingestão de fibra em detergente neutro. As digestibilidades aparentes e o metabolismo do nitrogênio não apresentaram efeitos significativos entre os tratamentos. No entanto, as emissões de CH4 foram significativamente inferiores nos grupos CPEG e SPEG em comparação com o grupo controle. Além disso, as expressões de populações microbianas de metanogênicas no grupo CPEG apresentaram tendência menor do que nos grupos SPEG e controle. As conclusões mais salientes do presente estudo foram que, usando planta rica em tanino em dietas de pequenos ruminantes, poderemos ter mais benefícios em termos de substituição da fonte de proteína da dieta (segurança alimentar) e redução da produção de CH4 entérico

Bifenilas policloradas (PCBs) são compostos de difícil degradação presentes na composição de ascarel, muito utilizado como fluidos dielétricos e isolantes. Neste contexto, a presente pesquisa teve como objetivo avaliar a diversidade de microrganismos em biofilmes de reatores anaeróbios na presença de PCB empregando Métodos de Microbiologia de Anaeróbios Estritos e de Biologia Molecular. Em reator anaeróbio horizontal de leito fixo (RAHLF), alimentado com etanol, formiato, Triton X-100 (0,1%) e ascarel (1 mL/L), operado com tempo de detenção hidráulica (TDH) de 24 horas, foi retirado a comunidade microbiana do biofilme da espuma de poliuretano. Os grupos microbianos encontrados por meio da clonagem e sequenciamento do gene RNAr 16S para o domínio Bacteria foram relacionados aos filos Thermogae, Proteobacteria (Brachymonas petroleovorans, 100% de similaridade e Methylobacillus, 98% de similaridade), Firmicutes (Clostridium, 97% de similaridade, Syntrophomonas, 100% de similaridade e Sporomusa com 100% de similaridade), Synegistetes (Synergistes, 98% de similaridade), Spirochaetes (Leptonema illini, 98% de similaridade), Aminanaerobia, Deferribacteres, Chlorobi, Chloroflexi e Armatimonadetes. Além disso, como nesse biofilme foram identificadas bactérias redutoras de ferro, procedeu-se a sua quantificação por meio da técnica de tubos múltiplos (NMP, Número Mais Provável) obtendo 5,26 x \'10 POT.12\' NMP/g STV de bactérias redutoras de ferro. Ensaio em batelada foi realizado separadamente sob duas condições: (1) metanogênica e (2) ferro redutora. Em ambas as condições foram adicionadas aroclor 1260 (PCB). Os reatores, sob condição metanogênica, foram alimentados com meio de cultivo Angelidaki e substratos orgânicos (formiato e etanol), além de aroclor 1260 (0,2 \'mü\'g/L). Para simular a condição redutora de ferro foi acrescido ao meio de cultura Angelidaki, EDTA férrico (1,86 g/L). A produção de metano, na presença de aroclor 1260 foi de 3,8 x \'10 POT.-4\' mmol \'CH IND.4\'/g STV. A presença de bactérias ferro redutoras foi confirmada indiretamente pela taxa média de redução férrica (90%) nos reatores em batelada, após 60 dias de operação. Por meio de PCR/DGGE, elaborou-se um dendograma das amostras deste ensaio em batelada (metanogênico e redutor de ferro) comparativamente com as do reator RAHLF (biofilme presente na parede do reator e no material suporte). Os reatores em batelada apresentaram similaridade entre si de 79% e 92% para os domínios Bacteria e Archaea, respectivamente. As amostras do reator RAHLF foram 80% (Bacteria) e 96% (Archaea) similares. A existência de bactérias degradadoras de PCB, bem como, bactérias redutoras de ferro no biofilme anaeróbio contribuiu com informações sobre o consórcio microbiano e sua diversidade. / Polychlorinated biphenyls (PCBs) are compounds of difficult degradation, a component of askarel, which were used widely as coolants and lubricants. Hence, this study evaluated the diversity of microorganisms in the presence of PCBs in anaerobic reactors. For such, methods as Strict Anaerobic Microbiology and Molecular Biology were employed. The microbial community of the biofilm, developed in a fixed horizontal bed anaerobic reactor (RAHLF), was studied using the technique of cloning and sequencing of RNAr 16S gene for the Bacteria domain. The reactor had immobilized cells in polyurethane foam with ethanol and formate as a carbon source, Triton X-100 (0.1%) and polychlorinated biphenyls (1 mL/L), and operated with 24 hours HRT. The microbial groups found in this biofilm were related to phyla Thermogae, Proteobacteria (Brachymonas petroleovorans, 100% similarity and Methylobacillus, 98% similarity), Firmicutes (Clostridium, 97% similarity Syntrophomonas, and 100% similarity with Sporomusa 100% similarity), Synegistetes (Synergistes, 98% similarity), Spirochaetes (Leptonema Illini, 98% similarity), Aminanaerobia, Deferribacteres, Chlorobi, Chloroflexi and Armatimonadetes. Furthermore, as bacteria that reduce iron were found, we proceeded the quantification by the multiple tube method (MPN) for this group, obtaining 5.26 x \'10 POT.12\' MPN/g STV of iron-reducing bacteria. The batch reactors evaluated the growth of microorganisms in two condictions: (1) methanogenic e (2) iron reduction, both had the presence of PCBs (Aroclor 1260). The reactor, under methanogenic condition, was fed with synthetic substrate Angelidaki, ethanol and formate, used as carbon source, and aroclor 1260 (0.2 \'mü\'g /L). To simulate the condition of iron reducing, the same synthetic substrate was supplemented with ferric EDTA (1.86 g/L). The production of methane in the presence of aroclor 1260, was 3.8 x \'10 POT.-4\' mmol \'CH IND.4\'/g STV. The presence of iron reducing bacteria, after 60 days, was confirmed indirectly by the average rate of iron ferric reduction (90%). Filogenetics analysis (PCR/DGGE) compared the samples of this batch reactor - methanogenic and reduction of iron ferric -, with the samples of RAHLF - the biofilm in the reactor wall and the support material. The two condictions in batch reactors showed similarity of 79% and 92% respectively for the Bacteria and Archaea domain. Therefore, both samples of RAHLF showed 80% (Bacteria) and 96% (Archaea) of similarity. In other words, more similarity were presented due configuration of the reactor as well as the type of PCB added. As a result, the existence of PCBs degrading bacteria and iron-reducing bacteria in anaerobic biofilm, provided informations about the microbial consortium and its diversity in the presence of PCB.

A metilamina ('CH IND.3'NH IND.2') é um composto orgânico usado na produção de inseticidas, herbicidas, fungicidas, surfactantes, combustíveis fósseis, explosivos, produtos farmacêuticos, químicos fotográficos, tintas, tecidos, solventes, borrachas e anti-corrosivos. Estudos sobre tratamento de águas residuárias contendo metilamina são escassos e se restringem aos trabalhos envolvendo pesticidas carbamatados. Visando contribuir com os estudos acerca da degradação anaeróbia da metilamina, esta pesquisa estudou a comunidade microbiana e a atividade metanogênica específica em reatores anaeróbios em batelada, inoculados com lodo granular oriundo de reator UASB usado no tratamento de água residuária de abatedouro de aves, sob diferentes condições nutricionais: controle – sem metilamina, 5 mM, 10 mM, 20 mM, 30 mM, 50 mM, 75 mM e 90 mM de metilamina. Os reatores foram incubados sob temperatura de 30°C e agitação de 150 rpm. Desses reatores foram obtidas amostras para a determinação da atividade metanogênica específica (AME), sólidos suspensos voláteis (SVT), nitrogênio amoniacal e exames microscópicos. Ao final do experimento, foram realizados exames da biomassa por meio da técnica do número mais provável (NMP) e análise da diversidade microbiana por PCR/DGGE e seqüenciamento. O aumento da AME foi proporcional ao aumento das concentrações de metilamina, com inibição de produção de metano apenas nos reatores alimentados com 90 mM de metilamina. Os reatores alimentados com 50 mM e 75 mM de metilamina apresentaram os melhores resultados, com valores médios de AME de 0,0804 mmol 'CH IND.4'/g SVT.h e 0,0825 mmol 'CH IND.4'/g SVT.h respectivamente. Nos exames microscópicos foi verificado semelhança de morfologias microbianas em todas as concentrações de metilamina estudadas. Os organismos presentes nos reatores foram Methanosarcina sp., Methanosaeta sp., bacilos, coco-bacilos, filamentos e cocos. Em relação à análise de DGGE, não houve variação significativa nos padrões de bandas, tanto para o domínio Archaea quanto para o domínio Bacteria. Com os resultados da técnica de número mais provável (NMP) observou-se a predominância de arquéias metanogênicas dentre as bactérias anaeróbias totais. / The methylamine ('CH IND.3'NH IND.2') is an organic compound used in the production of insecticides, herbicides, fungicides, surfactants, fossil fuels, explosives, pharmaceuticals, photographic chemicals, paints, textiles, dyes, rubber and anticorrosive chemicals. Some studies about the treatment of wastewater containing methylamine are scarce and limited to works involving carbamate pesticides. This research aimed to study the anaerobic degradation of methylamine, the microbial community and the specific methanogenic activity in anaerobic battled reactors. The reactors were inoculated with granular sludge from a UASB reactor treating poultry wastes. Different nutritional conditions were adopted in the operation of the reactors: control (without methylamine), 5 mM, 10 mM, 20 mM, 30 mM, 50 mM, 75 mM and 90 mM of methylamine. The reactors were incubated under standard conditions: 30ºC and 150 rpm. Samples had been removed from the reactors to determine the specific methanogenic activity, the concentration of volatile suspended solids and ammoniacal nitrogen and the microscopic analysis. At the end of the experiment, the biomass was studied by the most probable number (MPN) technique and by the microbial diversity analysis with PCR and DGGE techniques. The increase of the specific methanogenic activity was proportional to the increase of methylamine concentration. The methane production was inhibited only in the reactor that was fed with 90 mM of methylamine. The reactors that were fed with 50 mM and 75 mM of methylamine showed the best results, with medium values of specific methanogenic activity equal to 0,0804 mmol 'CH IND.4'/g SVT.h and 0,0825 mmol 'CH IND.4'/g SVT.h, respectively. The microscopic analysis showed similarity between the microbial morphologies in all of the reactors. The observed microorganisms were Methanosarcina sp., Methanosaeta sp., rods, cocci and filaments. The DGGE analysis did not show significant variation in the standard profile of the Archaea and Bacteria domains. The results of the MPN technique revealed the predominance of the methanogenic archaea among the total anaerobic bacteria.

Anaerobic Digestion (AD) is an essential component in wastewater treatment to recover energy from waste and deals with sludge management issues effectively. AD is a treatment process that converts organic matter to methane and carbon dioxide with multi-step biological reactions. Methanogenesis, the subprocess of AD that produces methane, is an important indicator of the stability of AD and is influenced by pH, temperature, ammonia, volatile fatty acids (VFAs), and solids concentrations among other factors. Ammonia is an essential nutrient for methanogenic bacteria but at certain ammonia concentrations and pH levels, ammonia is said to be a toxicant for methanogenic archaea. Substrates that are high in ammonia content can include those high in protein, such as food waste, which can be inhibitory to methanogens in the digestion process. Thickened waste activated sludge (TWAS) also contains a large amount of nitrogen with its higher solids concentration, promoting methane production. VFAs are produced during acidogenesis and they can negatively affect methanogenic archaea. High organic loading rates into AD can lead to an accumulation of VFAs and thus inhibition of methanogenic activity. Even with well-known inhibitory effects of ammonia and VFAs on methanogenesis, there are limited tools available for modelling these inhibitions, especially when evaluating diverse compositions of substrate. The objectives of this research work are to experiment for various pairings of pH, ammonia, and acetate levels using batch reactors and to quantify the inhibition on the overall methane production using an AD-based model focused on biological reactions. / Thesis / Master of Applied Science (MASc)

The objective of this study was to utilize an alternating anaerobic/aerobic sequence to biologically transform perchloroethylene to non-hazardous end products such as ethylene, CO2 and H2 using a single microbial consortia in a methanogenic and/or a homoacetogenic environment followed by a aerobic methanotrophic environment. Reductive dechlorination of PCE and TCE to cDCE and VC in an anaerobic environment is typically carried out by methanogens, sulfidogens, or homoacetogens but often (e.g. in-situ) leads to an accumulation of daughter compounds (cDCE, VC) which are more toxic than their parent compounds (PCE, TCE). Furthermore, PCE is resistant to degradation in aerobic environments while VC and cDCE are readily oxidized co-metabolically by aerobic methanotrophic bacteria, among others. In order to achieve complete mineralization of chlorinated solvents using a biotic system, an anaerobic/aerobic treatment strategy was investigated. This strategy has been accomplished successfully at a lab scale with anaerobic and aerobic reactors in series, and in-situ anaerobic zones with downgradient aerobic zones have been proposed in the field. In contrast, the focus of this research was to expose single mixed microbial consortia to sequential anaerobic/aerobic treatments in order to determine if reductive dechlorination could be sustained following aerobic phases of treatment. If possible this would imply that the anaerobic and aerobic zones (in-situ) or reactors (ex-situ) would not necessarily have to be spatially separated. In pure or dilute cultures where soil material is not present strict anaerobes would typically not resume metabolic activity if exposed to frequent aerobic phases of treatment. However in aquifer material or reactors with large floc/granules it might be possible due to the protection of anaerobic micro-environments as a result of diffusion limitations. Microcosms contained in sealed 120-mL serum bottles were used to generate experimental data including autoclaved abiotic controls with mercuric chloride. Inocula for these microcosms come from a several sources, including anaerobic digester sludge, soils, and contaminated aquifers. Once an experimental microcosm showed signs of reductive dechlorination, an aerobic treatment was implemented. The anaerobic phase of the microcosm was interrupted with a short duration aerobic phase. Headspace air or hydrogen peroxide addition was used to supply oxygen. Analytical data from the experiments indicated that anaerobic reductive dechlorination was readily accomplished during anaerobic phase experiments as PCE was sequentially dechlorinated to TCE and then to cDCE as reported in previous research reported by others in the literature. Additionally, a few mixed consortia microcosms showed evidence of further reductive dechlorination to VC and ethylene. During the sequential environment experiments, analytical data also indicated that reductive dechlorination also resumed after an aerobic sequence utilizing hydrogen peroxide as an oxidizer in the microcosm. No conclusive evidence was observed to indicate that aerobic degradation of cDCE during any of the aerobic phase treatments. This was probably due to the inocula not containing methanotrophs.

The anaerobic treatment of waste water has been studied with an emphasis on the Up- flow Anaerobic Sludge Blanket (UASB) reactor. A model to describe the processes occurring in a UASB reactor was developed and an experimental study of the anaerobic wastewater treatment systems in Nicaragua was also performed. Experimental work was carried out in order to link the study to the wastewater treatment situation in Nicaragua, a developing country. In order to assess the performance of the treatment plants, the methanogenic activity of sludge from seven anaerobic wastewater treatment plants was first addressed. Due to a lack of Standards for the measurement of methanogenic activity, a laboratory method was developed based on the methods found in the literature. An additional aim of this study was to find adequate inoculum for the wastewater treatment plant in a brewery using an anaerobic reactor. Physic-chemical characteristics of the sludge were also determined to provide a basis for decisions regarding the agricultural employment of the sludge from the treatment plants. A one-dimensional model describing the physical and biological processes occurring in an Up-flow Anaerobic Sludge Blanket reactor has been developed. These processes are advection, dispersion and reaction in the granule, including mass transport at the interface and diffusion within the particle. The advection-dispersion equation is used to describe these phenomena in the reactor. Dispersion is mainly caused by the gas bubbles rising up through the reactor and the granules in the ascending flow. The extent of the dispersion is expressed by the dimensionless Peclet (Pe) number. It is assumed that the biological degradation takes place at the surface and within the granules. The processes occurring in the granules formed by the microorganisms are described in detail; they include diffusion in the stagnant film around the granule, diffusion within the particle, and a degradation reaction. From these processes, the reaction term is analytically determined. The granules were modelled as spherical porous biocatalysts of different sizes. The biochemical degradation reactions were assumed to follow Monod type kinetics of the first order. For the numerical solution of the model, a standard program was used (Within MATLAB). The model was applied to some experimental data taken from the literature. An important characteristic of the model is that it can simultaneously take into account reactions in granules of different sizes. At present, the parameters of the model are calculated using data from the literature; but experimental measurements of the main parameters are planned. The impact of the different parameters was studied by numerical simulation and its validity was tested using experimental data reported in the literature. The model could be a useful tool in the performance optimization of UASB reactors by predicting the influences of different operational parameters. / QC 20101116

A series of lab-scaled digestion studies including conventional mesophilic anaerobic digestion(MAD), thermophilic anaerobic digestion (TAD) at a range of treatment temperatures, and mesophilic high solids digestion of thermally pretreated wastewater sludge (THD) were carried out. Enhanced digestion performance in terms of solids destruction and methane generation by THD relative to MAD was achieved, and was largely attributable to the solubilization and subsequent biodegradation of energy-rich substrates within blended primary and secondary sludge. TAD was observed to underperform MAD, especially at elevated temperatures as methanogenic inhibition resulted in the accumulation of headspace hydrogen, thus resulting in poor removal of volatile fatty acids. The thermodynamics of fatty acid metabolism was favorable at each digestion temperature, thus it was concluded that microbial inhibition was the controlling factor in poor thermophilic performance. Inhibition by free unionized ammonia (NH₃) was characterized for THD and MAD biomass. Acetic acid degradation was equally affected over a range of NH₃ concentrations; however, methane generation by THD was less sensitive to ammonia inhibition, thus suggesting that methanogenesis by THD was less dependent on the NH₃-sensitive process of aceticlastic methanogenesis. Total ammonia nitrogen (TAN) and bicarbonate alkalinity were stoichiometrically produced from proteinaceous material during thermal hydrolytic pretreatment and subsequent high solids anaerobic digestion. Combined effects of TAN and high pH resulted in NH₃-inhibition during THD. Kinetic evaluations suggested that a growth rate reduction of approximately 65% was associated with in-situ NH₃ concentrations of the THD reactor. NH₃-inhibition was apparently responsible for a shift in dominant methanogenic community of the aceticlastic Methanosarcina barkeri in MAD to the hydrogenotrophic Methanoculleus bourgensis in THD. A similar shift in methanogenic community was observed between low temperature thermophilic digestion at 47°C, where the dominant order was Methanosarcinales, to high temperature thermophilic digestion at 59°C where the dominant order was Methanobacteriales. These findings support a process-driven pathway shift from aceticlastic to non-aceticlastic methanogenesis between 180 and 290 mg/L NH₃-N. Such a threshold is supported by previous literature related to ammonia tolerance of pure cultures of methanogens and has significant implications for the kinetic design of advanced anaerobic digestion processes. / Ph. D.

Foram operados dois reatores UASB de bancada (volumes de 39,0 e 10,5 L) instalados em série, alimentados com águas residuárias de suinocultura com concentração de sólidos suspensos totais (SST), no primeiro reator, em torno de 5000 mg/L, com temperatura controlada (de 25 a 30 graus Celsius) e com tempo de detenção hidráulica (TDH) no primeiro reator de 62 a 16 h e no segundo de 16 a 4 h. O objetivo foi avaliar o desempenho e caracterizar física e microbiologicamente o lodo dos reatores UASB operados em dois estágios. Os resultados obtidos no ensaio 1, com TDH de 62 h no primeiro reator e SST no afluente de 5240 mg/L, mostraram eficiências de remoção de DQO total de 86% e 59% e SST de 82% e 57%, no primeiro e segundo reatores, respectivamente. A eficiência de remoção de DQO total e SST no sistema, no ensaio 1, foi de 95% e 94%. No ensaio 2, com TDH de 31 h no primeiro reator e SST de 5000 mg/L no afluente, observou-se eficiência de remoção de DQO total de 86% e 43% e SST de 85% e 58%, no primeiro e segundo reatores, respectivamente. A eficiência de remoção de DQO total e SST no sistema, no ensaio 2, foi de 92% e 94%. No ensaio 3, com TDH de 16 h no primeiro reator e SST de 5490 mg/L no afluente, observou-se eficiência de remoção de DQO total de 73% e 23% e SST de 65% e 20%, no primeiro e segundo reatores, respectivamente. A eficiência de remoção de DQO total e SST no sistema, no ensaio 3, foi de 79% e 73%. A TCOV aplicada no primeiro reator, no ensaio 1, foi de 4,55 kg DQO total/\'M POT.3\'.d, no ensaio 2 de 8,75 kg DQO total/\'M POT.3\'.d e no ensaio 3 de 18,65 kg DQO total/\'M POT.3\'.d. A produção de \'CH IND.4\' no primeiro reator foi de 17,50 a 68,20 L \'CH IND.4\'/d e no segundo reator de 1,62 a 5,50 L \'CH IND.4\'/d com a diminuição do TDH. Os reatores UASB instalados em série foram eficientes na remoção da fração dissolvida e, principalmente, da fração devido à concentração de SST do afluente. Para TCOV de 4,55 kg DQO/\'M POT.3\'.d no primeiro reator, pôde-se obter eficiências de remoção de DQO total e de SST acima de 90% e de DQO dissolvida acima de 85%. Para TCOV de 18,65 kg DQO/\'M POT.3\'.d no primeiro reator, as eficiências de remoção de DQO total e de SST foram acima de 70% e DQO dissolvida acima de 75%. As maiores produções específicas de metano foram obtidas com TCOV de 2,55 kg DQO/\'M POT.3\'.d para o segundo reator e de 8,65 kg DQO/\'M POT.3\'.d para o primeiro reator. A operação dos reatores UASB com valores de concentração de SST no afluente em torno de 5000 mg/L foram prejudiciais ao processo de granulação do lodo. Os grânulos apresentaram distribuição dispersa das morfologias microbianas ao longo da parede, não caracterizando a divisão em camadas definidas. As arqueas metanogênicas predominantes foram as semelhantes à Methanosaeta. / Two bench scale UASB reactors (volumes of 39,0 and 10,5 L) were operated in sequence, fed with swine wastewater with total suspended solids (TSS) concentration around 5000 mg/L in the first reactor, with controlled temperature (from 25 to 30 Celsius degrees) and operating with hydraulic detention time (HDT) in the first reactor varying from 62 to 16 h and in the second reactor from 16 to 4 h. The objective was to evaluate the performance and to characterize physically and microbiologically the sludge from UASB reactors operated in two stages treating swine wastewater. The results obtained in phase 1, with HDT of 62 h in the first reactor and TSS in the influent of 5240 mg/L, presented total COD removal efficiencies of 86% and 59% and TSS reduction efficiency of 82% and 57%, in the first and second reactors, respectively. The removal efficiency of total COD and TSS in the system, in phase 1, was 95% and 94%, respectively. In the phase 2, using HDT of 31 h in the first reactor and TSS of 5000 mg/L in the influent, it was observed a total COD removal efficiency of 86% and 43% and TSS reduction efficiency of 85% and 58%, in the first and second reactors, respectively. The system removal efficiency of total COD and TSS in phase 2, was 92% and 94%, respectively. In phase 3, with a HDT of 16 h in the first reactor and TSS of 5490 mg/L in the influent, it was observed a total COD removal efficiency of 73% and 23% and TSS reduction efficiency of 65% and 20%, in the first and second reactors, respectively. The total COD removal efficiency and TSS reduction efficiency in the system, in phase 3, was 79% and 73%, respectively. The volumetric organic loading rate (VOLR) applied in the first reactor, in phase 1, was 4,55 kg total COD/\'M POT.3\'.d, in phase 2 was 8,75 kg total COD/\'M POT.3\'.d and in phase 3 was 18,65 kg total COD/\'M POT.3\'.d. The \'CH IND.4\' production in the first reactor was from 17,50 to 68,20 L \'CH IND.4\'/d and in the second reactor from 1,62 to 5,50 L \'CH IND.4\'/d decreasing the HDT in the experiment phases. The UASB reactor installed in sequence were efficient in the dissolved fraction removal and, mainly, to the fraction due to the TSS influent concentration. The total VOL value of 4,55 kg COD/\'M POT.3\'.d was measured in the first reactor, it was achieved TSS and total COD removal efficiencies above 90% and dissolved COD above 85%. For the OVL of 18,65 kg COD/\'M POT.3\'.d in the first reactor the TSS and total COD removal were above 70% and dissolved COD above 75%. The larger methane specific production was obtained with a total VOL of 2,55 kg COD/\'M POT.3\'.d in the second reactor and 8.65 kg COD/\'M POT.3\'.d in the first reactor. The UASB reactors operation with the TSS concentration values of 5000 mg/L in the influent was prejudicing the sludge granulation process. The granules present a microbial morphology disperse distribution that doesn\'t characterize a layers defined distribution. The predominant metonogenic archeas were similar to Methanosaeta.

O estudo sobre a comunidade microbiana de um sistema de tratamento biológico de águas residuárias é de particular interesse, uma vez que o conhecimento da microbiologia do processo pode levar ao aperfeiçoamento de projetos e ao aumento da eficiência dos sistemas. Este trabalho avaliou a atividade microbiana, particularmente a metanogênica, na lagoa de estabilização anaeróbia da estação de tratamento de esgotos sanitários do município de Cajati - SP. Para isso adotou a taxonomia polifásica na caracterização dos microrganismos e de seus aspectos funcionais, buscando o conhecimento da diversidade dos microrganismos e suas relações nesse tipo de sistema anaeróbio. Objetivou também contribuir para o estabelecimento de um protocolo seguro na realização dos ensaios de atividade metanogênica específica (AME). Os estudos foram realizados com amostras dos sedimento da lagoa coletadas em três períodos diferentes, a saber: outubro/2003, outubro/2004 e dezembro/2004. Durante as amostragens foram determinadas variáveis abióticas como temperatura, condutividade, pH, potencial de óxido-redução e teor de oxigênio dissolvido. Medidas do conteúdo de sólidos totais e voláteis (SV) foram também realizadas. A avaliação da atividade microbiana foi feita por exames microscópicos de contraste de fase e fluorescência, AME, determinação do DNAtotal, FISH - hibridização in-situ com sondas fluorescentes e DGGE - eletroforese em gel com gradiente linear de agentes desnaturantes. Também procedeu-se a contagem de protozoários e análise da presença de algas e cianobactérias. Os resultados revelaram que ocorreu variação nas condições do processo biológico nos períodos amostrados, sendo que em outubro/2004, durante período de fortes chuvas e ventos, a eficiência na redução da DBO foi apenas 18,2%. Nesse período, constatou-se organismos como algas do gênero Chorella sp e cianobactérias do gênero Merismopedia sp. Nas demais coletas a remoção da matéria orgânica medida em DBO foi superior a 80%, com boa atividade anaeróbia. Os resultados mostraram que a relação So/Xo de 0,25 gDQO/g SV foi a mais adequada para determinar o valor de AME, e os ensaios com as amostras de outubro/2003 e dezembro/2004 revelaram valores de AME na faixa de 0,85 a 0,21 mg\'CH IND.4\'/gSV.d. Constatou-se a ocorrência de alterações na estrutura da comunidade microbiana inicial em relação à final do experimento de AME, por meio do DGGE. Verifcou-se também nesses ensaios, que o conteúdo de SV inicial variou entre amostras e substratos, conferindo alta variabilidade ao teste. Os perfís de DGGE das amostras coletadas revelaram variação na estrutura das comunidades microbianas no sedimento, e maior diversidade de bactérias e arquéias quando a lagoa anaeróbia apresentava boa eficiência na redução da DBO. A técnica FISH como adotada não foi eficaz para quantificar e identificar os microrganismos devido ao excesso de hibridizações inespecíficas. Mesmo com suas limitações, a técnica FISH revelou a presença de microrganismos dos Domínios Bacteria e Archaea. Nesse caso, a Família Methanobacteriaceae, a ordem Methanomicrobiales e o gênero Methanosaeta sp. foram confirmados. Nas diferentes coletas, foram identificados protozoários dos gêneros Paramecium sp. e Vorticella sp., e rotíferos dos gêneros Brachionus sp., Trichocerca sp., Synchaeta sp. e Keratella sp. / Microbial diversity studies have remarkable relevance since the knowledge about the microbiology of process can improve plants and system efficiency. This work assessed microbial activity, specially methanogenic, at anaerobic pond for domestic wastewater treatment in the city of Cajati, São Paulo State, Brazil. Poliphasic taxonomy was adopted in onder to contribute to the understanding of microbial community diversity and functionality. As well as to contribute for the establishment of a protocol to the specific methanogenic activity test (SMA). Three periods of sampling were done at the sediment of the anaerobic pond (october 2003, october 2004 and december 2004). Abiotic variables as temperature, conductivity, pH, dissolved oxygen and redox potential were measured at sampling time. TS and VS contents were determined in the samples. Microbial studies were done by observation on optical and fluorescent microscopy, analyse of SMA, totalDNA quantitation, counting of protozoa, analyze of algal and cyanobacteria presence, as well as application of two molecular techniques: Fluorescent in-situ Hybridization (FISH) and Denaturing Gradient Gel Electrophoresis (DGGE). Results showed that general conditions of the anaerobic pond changed among samplings. On October 2004, when the rain and wind were very strong, the organic matter removal efficiency (BOD basis) in the anaerobic pond was low (18.2%) and predominant microorganisms were of aerobic algae, as Chorella sp, and the blue-green algae Merismopedia sp. On the other hand, the removal of organics at other two samplings was more than 80% and the anaerobic microbial activity was verified in the sample. SMA tests showed the food/microorganism rate (F/M) of 0.25 gDQO/g VS was the most suitable to the samples. The results showed SMA values between 0.85 and 0.21 mg\'CH IND.4\'/gVS.d for samples of October 2003 and October 2004. SMA test induced modifications in the structure of the microbial community according to DGGE-profile. In addition, VS content, which was used in the SMA tests as biomass measurement, displayed variable behavior making test results difficult to interpret in some situations. DGGE-profile showed variation in the sediment community structure. Higher bacterial and archaeal diversity was observed when anaerobic pond showed 80%, or more, of DBO removal. FISH technique was not a suitable method to secure quantification and identification of the microorganisms from in excess. In spite of the technique limitations, it was possible to identify microorganisms of Bacteria Domain, Archaea Domain, Methanobacteriaceae Family, Methanomicrobiales Order, and microorganisms belonging to Methanosaeta genus. Paramecium and Vorticella were the protozoans identified in all samplings. Rotifers belonging to genders Brachionus, Trichocerca, Synchaeta and Keratella were also observed.