A atividade metanogênica específica foi estudada neste trabalho a partir do levantamento de diversas metodologias propostas para a realização desse teste . Procurou-se otimizar o teste de atividade metanogênica tomando-se por base a metodologia adotada por Willem de Zeeuw (1984). No desenvolvimento do trabalho experimentou-se várias condições metodológicas para o teste de atividade, objetivando-se a máxima atividade metanogênica específica do lodo, através de um ensaio que fosse simples e rápido. Para isso, utilizando-se de cinco tipos de lodos anaeróbios, estudou-se a influência das quantidades de biomassa e de substrato, da natureza do substrato, da forma de agitação do meio, das soluções de nutrientes e de metais, da redução do meio com nitrogênio, do volume dos frascos-reatores e do tempo de armazenamento do lodo nos resultados dos testes de atividade. Observou-se que, em função do tipo de lodo e da sua atividade metanogênica, devem ser pesquisados o substrato mais adequado e a relação ótima entre a quantidade de substrato e biomassa que conduzam à atividade metanogênica específica máxima durante o teste. Observou-se também que o ensaio pode ser mais simples e mais rápido, comparativamente àquele baseado na metodologia de de Zeeuw (1984). Como conclusão principal são apresentadas novas condições metodológicas para o teste de atividade metanogênica específica para lodos anaeróbios. / The specific methanogenic activity test was studied with basis on several methodologies proposed for carrying out the referred test. The methodology used by Willem de Zeeuw (1984) was employed to optimize this test. Several methodological processes were used for the activity test in an attempt to reach the maximum specific methanogenic activity of the sludge in a simple and quick process. Five types of anaerobic sludges were used and the following parameters were studied: the influence of the amounts of biomass and substrate, the nature of substrate, the liquor mixing, the nutriente and metal solutions, oxygen removal the reactor capacity, and the sludge storage time in the results of the activity tests. It was observed that the more suitable substrate and the optimum relationship between the quantity of substrate and biomass must be researched, sludge, so that the referred according to the type of relationship leads to a maximum specific methanogenic activity. It was observed that the test can be quicker and simpler than de Zeeuw\'s (1984). The main result reached consists of the presentation of new conditions to carry out the test of specific methanogenic activity for anaerobic sludges.

In theoretic parts diploma work is described mechanism anaerobic of the process, factors which him work and description qualitative determination biogas. Objective practical parts was quantification and qualities produced biogas. Led I'm investigation intent on investigation choice type phytomass and their fitness for production and quality biogas. On fermentative sweetness I'm watched change pH, Cox, decrease organic masses along anaerobic digestion. Further I'm establish entitlements content lignin and CHSK in single figures phytomass. On device OxiTop( control AN12 I'm measured maximum yield and speed development methane and biogas on hundreddweight substrates

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.

<p>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.</p><p>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.</p><p>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.</p><p>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.</p>

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.

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

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.

Made available in DSpace on 2014-06-11T19:32:54Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-02-24Bitstream added on 2014-06-13T20:24:21Z : No. of bitstreams: 1 duda_rm_dr_jabo.pdf: 7705565 bytes, checksum: 22edf4cc4e57fedc9fa2426e83e181e0 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Avaliou-se o desempenho de um reator anaeróbio de fluxo ascendente com manta de lodo (UASB) seguido de um filtro anaeróbio de fluxo ascendente, instalados em série, com volume total de 300 L e 190 L, respectivamente, no tratamento de águas residuárias de suinocultura. As cargas orgânicas volumétricas aplicadas no reator UASB foram de 9,2 a 26,7 g DQOtotal (L d)-1. Para o pós-tratamento do efluente do sistema de tratamento anaeróbio utilizou-se um filtro biológico percolador (FBP) com volume total de 250 L. O meio suporte utilizado no filtro anaeróbio e no FBP foi composto por anéis de bambu e anéis de eletroduto plástico corrugado. As cargas hidráulicas superficiais (Qs) aplicadas no FBP variaram de 3,5 a 21,1 m3 (m2 d)-1. Os tempos de detenção hidráulica (TDH) aplicados no sistema de tratamento anaeróbio e pós-tratamento foram de 23,3 a 66,6 h. As produções volumétricas de metano no sistema de tratamento anaeróbio variaram de 0,16 a 0,68 L CH4 (L reator d)-1. Foram observadas eficiências médias de remoção de até 97% para a demanda química de oxigênio total, de 98% para os sólidos suspensos totais, de 78% para o nitrogênio total, de 79% para o fósforo total, de 99,9% para o cobre, de 99,9% para o zinco e de 99,99% para os coliformes termotolerantes, no sistema de tratamento anaeróbio e póstratamento. Com a utilização de diferentes fontes de substrato no ensaio de atividade da microbiota, observou-se o crescimento equilibrado das populações hidrolíticas, acidogênicas, acetogênicas e metanogênicas acetoclásticas e hidrogenotróficas no lodo dos reatores anaeróbios. Com a aplicação da técnica de biologia molecular (metagenômica) foram identificadas no lodo do reator UASB e do filtro anaeróbio de fluxo ascendente arquéias metanogênicas das famílias Methanomicrobiaceae, Methanobacteriacea, Methanosaetaceae e Methanosarcinaceae. / The performance of an upflow anaerobic sludge blanket (UASB) followed of the anaerobic filter, installed in series, were evaluated for the treatment of swine wastewater. The total volume of UASB and anaerobic filter were of 300 L and 190 L, respectively. The organic load rate applied on the reactor UASB were of 9.2 to 26.7 g total COD (L d)-1, in the start and assays 1, 2, 3 and 4. For the post-treatment of effluent the anaerobic system was used a trickling filter, with total volume of 250 L. The supports used in the anaerobic filter and trickling filter were composed by bamboo rings in the start up and assays 1 to 2 and plastics rings in the assays 3 to 4. The hydraulic load applied on the trickling filter were of 3.5 to 21.1 m3 (m2 d)-1. The hydraulic detetion time (HDT) applied of system anaerobic and post treatment were 23.3 at 66.6 h. The volumetric methane productions ranged from 0.16 a 0.68 L CH4 (L reactor d)-1. The efficiencies of removal the chemical oxygen demand, total solids suspended, copper, zinc and fecal coliforms were of up to 97, 98, 78, 79, 99.9, 99.9 e 99.99%, respectively, for the anaerobic and post-treatment. With the use of different substratum sources in the assays of specific methanogenic activity, it was observed the balanced growth of the populations of microorganisms in the sludge of anaerobic reactor. The archaeas of the families Methanomicrobiaceae, Methanobacteriaceae; Methanosaetaceae and Methanosarcinaceae were identified in the sludge of the reactor UASB and of the anaerobic filter.

Orientador: Roberto Alves de Oliveira / Banca: Eugênio Foresti / Banca: Maria Bernadete Amâncio Varesche / Banca: Manoel Victor Franco Lemos / Banca: Edson Aparecido Abdul Nour / Resumo: Avaliou-se o desempenho de um reator anaeróbio de fluxo ascendente com manta de lodo (UASB) seguido de um filtro anaeróbio de fluxo ascendente, instalados em série, com volume total de 300 L e 190 L, respectivamente, no tratamento de águas residuárias de suinocultura. As cargas orgânicas volumétricas aplicadas no reator UASB foram de 9,2 a 26,7 g DQOtotal (L d)-1. Para o pós-tratamento do efluente do sistema de tratamento anaeróbio utilizou-se um filtro biológico percolador (FBP) com volume total de 250 L. O meio suporte utilizado no filtro anaeróbio e no FBP foi composto por anéis de bambu e anéis de eletroduto plástico corrugado. As cargas hidráulicas superficiais (Qs) aplicadas no FBP variaram de 3,5 a 21,1 m3 (m2 d)-1. Os tempos de detenção hidráulica (TDH) aplicados no sistema de tratamento anaeróbio e pós-tratamento foram de 23,3 a 66,6 h. As produções volumétricas de metano no sistema de tratamento anaeróbio variaram de 0,16 a 0,68 L CH4 (L reator d)-1. Foram observadas eficiências médias de remoção de até 97% para a demanda química de oxigênio total, de 98% para os sólidos suspensos totais, de 78% para o nitrogênio total, de 79% para o fósforo total, de 99,9% para o cobre, de 99,9% para o zinco e de 99,99% para os coliformes termotolerantes, no sistema de tratamento anaeróbio e póstratamento. Com a utilização de diferentes fontes de substrato no ensaio de atividade da microbiota, observou-se o crescimento equilibrado das populações hidrolíticas, acidogênicas, acetogênicas e metanogênicas acetoclásticas e hidrogenotróficas no lodo dos reatores anaeróbios. Com a aplicação da técnica de biologia molecular (metagenômica) foram identificadas no lodo do reator UASB e do filtro anaeróbio de fluxo ascendente arquéias metanogênicas das famílias Methanomicrobiaceae, Methanobacteriacea, Methanosaetaceae e Methanosarcinaceae. / Abstract: The performance of an upflow anaerobic sludge blanket (UASB) followed of the anaerobic filter, installed in series, were evaluated for the treatment of swine wastewater. The total volume of UASB and anaerobic filter were of 300 L and 190 L, respectively. The organic load rate applied on the reactor UASB were of 9.2 to 26.7 g total COD (L d)-1, in the start and assays 1, 2, 3 and 4. For the post-treatment of effluent the anaerobic system was used a trickling filter, with total volume of 250 L. The supports used in the anaerobic filter and trickling filter were composed by bamboo rings in the start up and assays 1 to 2 and plastics rings in the assays 3 to 4. The hydraulic load applied on the trickling filter were of 3.5 to 21.1 m3 (m2 d)-1. The hydraulic detetion time (HDT) applied of system anaerobic and post treatment were 23.3 at 66.6 h. The volumetric methane productions ranged from 0.16 a 0.68 L CH4 (L reactor d)-1. The efficiencies of removal the chemical oxygen demand, total solids suspended, copper, zinc and fecal coliforms were of up to 97, 98, 78, 79, 99.9, 99.9 e 99.99%, respectively, for the anaerobic and post-treatment. With the use of different substratum sources in the assays of specific methanogenic activity, it was observed the balanced growth of the populations of microorganisms in the sludge of anaerobic reactor. The archaeas of the families Methanomicrobiaceae, Methanobacteriaceae; Methanosaetaceae and Methanosarcinaceae were identified in the sludge of the reactor UASB and of the anaerobic filter. / Doutor

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.