Made available in DSpace on 2017-05-12T14:47:38Z (GMT). No. of bitstreams: 1 Carla Limberger _Lopes (Tese revisada) 2016.pdf: 2793834 bytes, checksum: bd8e57da9e9986df4f212107c57ca249 (MD5) Previous issue date: 2016-02-17 / The aim of this study was to evaluate a combined anaerobic-aerobic upflow fixed-bed reactor with recirculation of the liquid phase for the removal of nitrogen and organic matter from poultry slaughterhouse wastewater. The reactor was made of an acrylic tube of internal diameter of 93 mm and the length of 1000 mm with a useful volume of 5.6 L being 3.5 L corresponding to the anaerobic compartments and 2.1 L to the aerobic one. The bed for immobilization of the biomass was formed by expanded clay and polyurethane foam. For discussing the results, this study was divided into three articles. In the first article, was evaluated the reactor performance with respect to the elimination of nitrogen (≈65 mg.NT.L-1) and organic matter (≈600 mg.DQO.L-1) due to the recirculation rate (R = 0.5, 1 and 2) and the hydraulic retention time (HRT) of 11 h (6.8 h in anaerobic condition and 4.2 h in aerobic condition) over the time. The best operating condition was obtained at the recirculating rate of 2. In this condition, the total nitrogen removal was 65% with the effluent concentration of 6 mg.NH4+.L-1 e 12 mg.NO3-.L-1. For all condition, the organic matter removal was greater than to 95% with the effluent concentration of approximately 20 mg.COD.L-1. Thus, the increasing of the recirculation rate influenced positively in the reactor performance. In the second article, the hydraulic detention time was evaluated (HDT) at 14 h, 11 h and 8 h with the recirculation rate (R) of 0.5 (Step I), R = 1 (Step II) and R 2 = (Step III). The affluent average concentrations were 65 mg.NT.L-1, 580 mg.COD.L-1, 77 mg L-1 of total alkalinity and pH of 6.4. The samples were collected inlet and output of each compartment along the reactor height. In the Step I, the nitrification efficiencies were 76%, 70% and 41% respectively for 14 h, 8 h and 11 h, showing the effect of HRT. In all steps, the alkalinity has been regarded as the limiting factor of the process and its deficit was 10 to 30%. It was attributed to this factor the low efficiency of total nitrogen removal of about 45%. Throughout the experiment the removal efficiency of organic matter in terms of chemical oxygen demand (COD) was over 90% and made to fit the first order kinetic model for degradation of the substrate. In the third article, was evaluated the hydrodynamic behavior of this system, through stimulus response tests using eosin Y dye as tracer. It has been evaluated the hydraulic retention time of 8 h with three recirculation rates, of .0.5, 1 and 2 times. Under these conditions, the removal of organic matter was greater than 90% and nitrogen conversion was favored by applying lower loads of (0.18 Kg.N.m-3.d-1). It was found that the hydrodynamic evaluation showed the continuous-stirred reactors behavior with 2 to 2.5 reactors in series (N-CSTR). / O objetivo desse trabalho foi avaliar um reator combinado anaeróbio-aeróbio de leito fixo e fluxo ascendente com recirculação da fase líquida, para a remoção de nitrogênio e matéria orgânica proveniente de água residuária de abatedouro de aves. O reator foi confeccionado em um tubo de acrílico de diâmetro interno de 93 mm e comprimento de 1000 mm, com volume útil de 5,6 L, sendo 3,5 L correspondentes aos compartimentos anaeróbios e 2,1 L correspondentes ao compartimento aeróbio. O leito para a imobilização da biomassa foi formado por argila expandida e espuma de poliuretano. A apresentação desse trabalho foi dividida em três artigos. No primeiro artigo, avaliou-se o desempenho do reator em relação à remoção de nitrogênio (≈65 mg.N.L-1) e de matéria orgânica (≈600 mg.DQO.L-1) em função da taxa de recirculação (R=0,5; 1 e 2) e do tempo de detenção hidráulica (TDH) de 11 horas (6,8 horas na condição anaeróbia e 4,2 horas na condição aeróbia), ao longo do tempo. A melhor condição operacional foi obtida com taxa de recirculação de 2. Nessa condição, a eficiência de remoção de nitrogênio total foi de 65% com concentrações efluentes de 6 mg.NH4+.L-1 e 12 mg.NO3-.L-1. Para todas as condições testadas, a eficiência de remoção de matéria orgânica apresentou-se superior a 95%, com concentração efluente de aproximadamente 20 mg.DQO.L-1. Assim, o aumento da taxa de recirculação influenciou positivamente no desempenho do reator. No segundo artigo, avaliaram-se os tempos de detenção hidráulica (TDH) de 14 horas, 11 horas e 8 horas com taxa de recirculação (R) de 0,5 (Ensaio I), R=1 (Ensaio II) e R=2 (Ensaio III). As concentrações médias afluentes foram 65 mg.NT.L-1, 580 mg.DQO.L-1, 77 mg.L-1 de alcalinidade total e pH de 6,4. As amostras foram coletadas na entrada e saída de cada compartimento, ao longo da altura do reator. Na Etapa I, e as eficiências de nitrificação foram de 76%, 70% e 41%, respectivamente para 14 horas, 11 horas e 8 horas, evidenciando o efeito do TDH. Em todas as etapas, a alcalinidade foi considerada o fator limitante do processo e o seu déficit variou de 10% a 30%. Atribuiu-se a esse fator a baixa eficiência na eliminação de nitrogênio total de aproximadamente 45%. Durante todo o experimento, a eficiência de remoção de matéria orgânica em termos de demanda química de oxigênio (DQO) foi superior a 90% e apresentou ajuste ao modelo cinético de primeira ordem para a degradação do substrato. No terceiro artigo, avaliou-se o comportamento hidrodinâmico desse sistema, a partir de ensaios de estímulo resposta utilizando o traçador Eosina Y. Avaliou-se o tempo de detenção hidráulica de 8 horas com as três taxas de recirculação, de 0,5, 1 e 2 vezes. Nessas condições, a remoção de matéria orgânica foi superior a 90% e a conversão de nitrogênio foi beneficiada com a aplicação de cargas menores (0,18 Kg.N.m-3.d-1). Na avaliação hidrodinâmica verificou-se que o reator apresentou comportamento de reator de mistura completa com 2 a 2,5 reatores em série (N-CSTR).

O presente trabalho constou da concepção de um reator anaeróbio em regime de batelada com circulação da fase aquosa contendo biomassa imobilizada em espuma de poliuretano. O reator foi operado em ciclos de 8 h e mantido a 30 ± 1ºC tratando água residuária sintética elaborada à base de glicose, com concentração de aproximadamente 500 mgDQO/L. O estudo hidrodinâmico realizado com velocidade superficial de recirculação entre 0,16 e 0,80 cm/s indicou que o tempo de mistura obtido em todas as condições pode ser considerado desprezível quando comparado ao tempo total do ciclo, e também permitiu considerar o comportamento do escoamento no leito como o de um reator pistonado. O desempenho do reator no tratamento da água residuária foi avaliado para a condição sem circulação e também para velocidades superficiais de recirculação entre 0,03 e 0,30 cm/s. Os resultados obtidos indicaram que o reator apresentou desempenho satisfatório e boa estabilidade, apresentando eficiências de até 96% de remoção da matéria orgânica. O aumento da velocidade superficial de recirculação diminuiu a resistência a transferência de massa na fase líquida, ocasionando um aumento de 115% na velocidade global de reação, estimada através do ajuste dos valores experimentais da concentração de substrato ao modelo de primeira ordem. / A new configuration of an anaerobic bioreactor with external circulation of the liquid phase wherein the biomass was immobilized on a polyurethane foam matrix is proposed. 8-hours cycles were cartied out at a temperature of 30 ± 1°C treating glucose synthetic wastewater at a concentration around 500 mg/L. A hydrodynamic study performed at 0.16 to 0.80 cm/s showed that the obtained mixture time is insignificant compared to the total cycles time, and the flow through the polyurethane foam bed can be represented by a plug flow. The reactor\'s performance assessed without circulation and with circulating liquid superficial velocity between 0.03 and 0.30 cm/s. The reactor attained operating stability and a COD removal efficiency of 96% was achieved. The increase in the liquid superficial velocity decreases the liquid resistance mass transfer, resulting in an increase of 115% in the global reaction velocity, estimated through the fit of a first model equation on the substrate concentration experimental values.

The reversible oxidation of cysteine by reactive oxygen species (ROS) is both a mechanism for cellular protein signaling as well as a cause of cellular injury and death through the generation of oxidative stress. The study of cysteine oxidation is complicated by the methodology currently available to isolate and enrich oxidized-cysteine containing proteins. We sought to simplify this process by reducing the time needed to process samples and reducing sample loss and contamination risk. We accomplished this by eliminating precipitation steps needed for the protocol by (a) introducing an in-solution NEM-quenching step prior to reduction and (b) replacing soluble dithiothreitol reductant with a series of newly-developed high-capacity polyacrylamide-based solid-phase reductants that could be easily separated from the lysate through centrifugation. These modifications, collectively called resin-assisted reduction and capture (RARC), reduced the time needed to perform the RAC method from 2-3 days to 4-5 hours, while the overall quality and quantity of previously-oxidized cysteines captured was increased. In order to demonstrate the RARC method’s utility in studying complex cellular oxidants, the optimized methodology was used to study cysteine oxidation caused by the redox-active metals arsenic, cadmium, and chromium. As(III), Cr(VI), and Cd(II) were all found to increase cysteine oxidation significantly, with As(III) and Cd(II) inducing more oxidation than Cr(VI) following a 24-hour exposure to cytotoxic concentrations. Label-free proteomic analysis and western blotting of RARC-isolated oxidized proteins found a high degree of commonality between the proteins oxidized by these metals, with cytoskeletal, translational, stress response, and metabolic proteins all being oxidized. Several previously-unreported redox-active cysteines were also identified. These results indicate that cysteine oxidation by As(III), Cr(VI), and Cd(II) may play a significant role in these metals’ cytotoxicity and demonstrates the utility of the RARC method as a strategy for studying reversible cysteine oxidation by oxidants in oxidative signaling and disease. The RARC method is a simplification and improvement upon the current state of the art which decreases the barrier of entry to studying cysteine oxidation, allowing more researchers to study this modification. We predict that the RARC methodology will be critical in expanding our understanding of reactive cysteines in cellular function and disease.

<p>When drug molecules are passively absorbed through the cell membrane in the small intestine, the first key step is partitioning of the drug into the membrane. Partition data can therefore be used to predict drug absorption. The partitioning of a solute can be analyzed by drug partition chromatography on immobilized model membranes, where the chromatographic retention of the solute reflects the partitioning. The aims of this thesis were to develop the model membranes used in drug partition chromatography and to study the effects of different membrane components and membrane structures on drug partitioning, in order to characterize drug–membrane interactions.</p><p>Electrostatic effects were observed on the partitioning of charged drugs into liposomes containing charged detergent, lipid or phospholipid; bilayer disks; proteoliposomes and porcine intestinal brush border membrane vesicles (BBMVs), and on the retention of an oligonucleotide on positive liposomes. Biological membranes are naturally charged, which will affect drug partitioning in the human body.</p><p>Proteoliposomes containing transmembrane proteins and cholesterol, BBMVs and bilayer disks were used as novel model membranes in drug partition chromatography. Partition data obtained on proteoliposomes and BBMVs demonstrated how cholesterol and transmembrane proteins interact with drug molecules. Such interactions will occur between drugs and natural cell membranes. In the use of immobilized BBMVs for drug partition chromatography, yet unsolved problems with the stability of the membrane were encountered. A comparison of partition data obtained on bilayer disks with data on multi- and unilamellar liposomes indicated that the structure of the membrane affect the partitioning. The most accurate partition values might be obtained on bilayer disks.</p><p>Drug partition data obtained on immobilized model membranes include both hydrophobic and electrostatic interactions. Such partition data should preferably be used when deriving algorithms or computer programs for prediction of drug absorption.</p>

Se presenta un modelo matemático para el diseño y simulación de un reactor de lecho fijo con enzimas inmovilizadas en partículas esféricas porosas. La ecuación de diseño del reactor se ha resuelto para el caso de un sistema enzimático, con limitaciones difusionales internas, que obedece a una cinética de Michaelis-Menten reversible.La validez del modelo se ha comprobado con el sistema enzimático naringina/naringinasa, aplicable al proceso de desamargado de zumos cítricos. / A mathematical model for design and simulation of a fixed bed reactor with immobilized enzymes in spherical particles is presented. The reactor design equation is solved for an enzymatic system taking into account internal diffusional limitations. Moreover, the enzyme obeys a reversible Michaelis-Menten kinetic. The validity of the model is checked by using the enzymatic system naringine/naringinase, which is used for fruit juice debbitering processes.

Cell adhesion to the extracellular matrix (ECM) provides tissue structure and integrity as well as triggers signals that regulate complex biological processes such as cell cycle progression and tissue-specific cell differentiation. Hence, cell adhesion is critical to numerous physiological and pathological processes, including embryonic development, cancer metastasis, and wound healing, as well as biotechnological applications, such as host responses to implanted devices and integration of tissue-engineered constructs. During the adhesion process, integrin surface receptors bind ECM proteins, cluster, and associate with the actin cytoskeleton. Subsequent strengthening of the integrin/actin cytoskeleton interaction occurs via complexes of proteins known as focal adhesions. Due to the close association between biochemical and biophysical processes within adhesion complexes, mechanical analyses can provide important new insights into structure/function relationships involved in regulating the adhesion process. The objective of this project was to investigate the role of the protein tyrosine kinase FAK in cell adhesion strengthening. Our central hypothesis was that FAK regulates adhesion strengthening by modulating interactions between integrins and FA structural components. Using a novel combination of genetically engineered cells to control the interactions of FAK, a spinning disk adhesion assay with micropatterned substrates to obtain reproducible and sensitive measurements of adhesion strength, and quantitative biochemical assays for analyzing changes in adhesive complexes, we demonstrate that FAK modulates adhesion strengthening via two distinct mechanisms: (1) FAK expression results in elevated integrin activation leading to regulation of strengthening rate and (2) FAK regulates steady-state adhesion strength via vinculin recruitment to focal adhesions. We also show that the autophosphorylation and catalytic sites of FAK are critical to this regulation of adhesion strengthening. This work is significant because it both identifies functional mechanisms of FAK and provides the first evidence that focal adhesion signaling regulates the adhesion strengthening process. Furthermore, this research demonstrates that the dependency of migration on adhesion strength is highly complex and establishes a need for adhesion strengthening metrics in analyzing the functional mechanisms of molecules within adhesion complexes.

Clostridium chauvoei is a gram-positive, spore-forming anaerobic bacterium. It is the pathogenic agent of blackleg, a disease causing serious toxemia and high mortality in cattle, sheep and many other domestic and wild animals. It is considered the most important Clostridium producing economic losses in livestock. Typically, animals infected with blackleg die rapidly without any signs of illness. Animals quickly die within 12 to 48 hours after contracting the disease. Therefore, the control of this disease is done by commercial vaccines consisting of whole formolized cultures. Immunity against C. chauvoei is associated with whole cell, including its somatic and flagellar antigens while in other clostridial diseases, protective immunity is obtained by the use of vaccines containing toxoids. Moreover, it is essential to obtain new information about the somatic antigens of C. chauvoei. Proteomics is the study of the proteome, the protein complement of the genome. The proteome has been defined as the entire complement of proteins expressed by a cell, organism, or tissue type, and accordingly, proteomics is the study of this complement expressed at a given time or under certain environmental conditions. 2-DE with Immobilized pH Gradients (IPGs) combined with protein identification by Mass Spectrometry (MS) is currently the workhorse for proteomics. Much of information about immunogenic component can be derived from proteomics coupled to Western blotting, namely immunoproteomics. Our study constitutes the first immunoproteomic analysis of C. chauvoei to identify candidate immunogenic antigens for development of new vaccines. Analyses were performed by Western blot and dot blot techniques against the whole cell extract proteins of C. chauvoei separated by 2-DE. Firstly, the growth conditions of two different strains, C. chauvoei ATCC 11957 and C. chauvoei 20 were optimized. After mice immunization studies with experimental vaccines prepared, sera were obtained for evaluation of the immunoglobulin G antibody level by ELISA. After high level of antibody response determination, 1-DE, 2-DE and immunoblot studies were performed for the characterization of immunogenic proteins. In the study, a total of 460 protein spots could be detected on the 2-DE gels by the help of Delta2D image analysis software and 30 of them were reacted with polyclonal antibodies against inactivated whole cells of C. chauvoei. Among these 30 spots, and 8 of them could be characterized by MALDI-TOF MS analyses. Of these 8 spots revealed four different gene products (distinct ORFs). Ornithine decarboxylase, methionine adenosyltransferase, glucose-6-phosphate isomerase, and flagellin protein FliB (C) are the characterized proteins. Glucose-6-phosphate isomerase has been identified as an immunogenic protein for a pathogenic microbe and in C. chauvoei for the first time. Methionine adenosyltransferase and ornithine decarboxylation were identified as immunogenic for C. chauvoei for the first time. The last defined protein is the flagellin protein FliB(C) which is known to be major immunogenic protein of C. chauvoei.

When drug molecules are passively absorbed through the cell membrane in the small intestine, the first key step is partitioning of the drug into the membrane. Partition data can therefore be used to predict drug absorption. The partitioning of a solute can be analyzed by drug partition chromatography on immobilized model membranes, where the chromatographic retention of the solute reflects the partitioning. The aims of this thesis were to develop the model membranes used in drug partition chromatography and to study the effects of different membrane components and membrane structures on drug partitioning, in order to characterize drug–membrane interactions. Electrostatic effects were observed on the partitioning of charged drugs into liposomes containing charged detergent, lipid or phospholipid; bilayer disks; proteoliposomes and porcine intestinal brush border membrane vesicles (BBMVs), and on the retention of an oligonucleotide on positive liposomes. Biological membranes are naturally charged, which will affect drug partitioning in the human body. Proteoliposomes containing transmembrane proteins and cholesterol, BBMVs and bilayer disks were used as novel model membranes in drug partition chromatography. Partition data obtained on proteoliposomes and BBMVs demonstrated how cholesterol and transmembrane proteins interact with drug molecules. Such interactions will occur between drugs and natural cell membranes. In the use of immobilized BBMVs for drug partition chromatography, yet unsolved problems with the stability of the membrane were encountered. A comparison of partition data obtained on bilayer disks with data on multi- and unilamellar liposomes indicated that the structure of the membrane affect the partitioning. The most accurate partition values might be obtained on bilayer disks. Drug partition data obtained on immobilized model membranes include both hydrophobic and electrostatic interactions. Such partition data should preferably be used when deriving algorithms or computer programs for prediction of drug absorption.

Made available in DSpace on 2014-06-11T19:23:27Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-02-26Bitstream added on 2014-06-13T19:09:12Z : No. of bitstreams: 1 covizzi_lg_me_sjrp.pdf: 1482206 bytes, checksum: 2f1c1f77dc261f160aba2bc3a1d1ffea (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O uso de células microbianas imobilizadas para aumentar a produção de metabólitos fúngicos em processos fermentativos tem mostrado altos rendimentos. Nesse trabalho foi avaliado pela primeira vez, a imobilização de células do Botryosphaeria rhodina, um fungo ligninolitico produtor constitutivo de lacases. Três suportes foram avaliados: Fibra Acrílica Fina (FAF); Espuma de Poliuretano Expandido (EPE); Espuma de Poliuretano Fibroso (EPF). O EPF foi o melhor suporte por ter mostrado uma imobilização mais homogenia das células. Um planejamento fatorial foi desenvolvido para otimizar a produção de lacases por células livres, na presença de álcool veratrílico (AV). A análise da superfície de resposta mostrou que 18mM como a melhor concentração de AV para a produção de lacases, usando-se 3 mL de homogeinato de células como inóculo (DOλ400nm 0.4-0.6) para 25 mL de meio de cultura em frascos de 125mL, a 180 rmp, durante 126 horas a 28ºC. O perfil de crescimento do fungo, associado a produção de lacase foram comparados na presença e na ausência de AV, usando-se células livres e células imobilizadas do B. rhodina. A imobilização aumentou aproximadamente 3 vezes a produção de lacases e manteve estável o nível de produção durante 6 reciclos. A imobilização de células do B. rhodina mostrou-se útil uma vez que economizou 72horas para atingir a maior produção de lacase, quando comparada com células livres e também aumentou a tolerância do fungo a concentrações mais altas de AV (500mM) / The use of microbial immobilized cells to increase the production of fungal metabolites in fermentation processes has showed higher yields. This work evaluated by the first time, the immobilization of Botryosphaeria rhodina cells, a ligninolytic fungus that produces laccase. Three carriers were evaluated: acrylic fine fiber (FAF), expanded polyurethane foam (EPE), and fiber polyurethane foam (EPF). The EPF was the best carrier because showed a homogeneous immobilization cells. A factorial design was developed in order to optimize the laccase production by free cells in the presence of the laccase inducer veratryl alcohol (VA). The analysis by response surface answer showed 18 mM as the best VA concentration to produce laccase using 3 mL of a cell homogenate (ODλ400nm 0.4-0.6) as inoculum, to 25 mL of culture medium in shaked flasks (125 mL) at 180 rpm, during 126 hours at 28 °C. A growth profile for laccase and fungal biomass production were compared with and without VA using free and immobilized cells of B. rhodina. The cell immobilization increased approximately 3 folds the laccase production and maintained it stable during 6 consecutive recycles. The cell immobilization of B. rhodina showed to be useful once saved 72 hours to achieve the higher laccase production when compared to the one with free cells, and also increased the fungal cell tolerance at higher VA concentrations (500 mM)

O presente trabalho teve como objetivo principal avaliar a performance e a estabilidade operacional de uma nova configuração de um reator anaeróbio híbrido (RAH) que combina um filtro anaeróbio de fluxo descendente seguido de uma câmara de manta de lodo de fluxo ascendente comparativamente ao reator UASB nas mesmas condições operacionais aplicados ao tratamento de esgoto sanitário. A avaliação da eficiência de remoção de remoção da matéria orgânica e dos sólidos suspensos nos reatores piloto foi por meio de testes da DQO, DBO, sólidos em suspensão totais (SST) e dos sólidos em suspensão voláteis (SSV).Os reatores UASB e RAH foram operados por um período de 240 dias na fase 1 e 150 dias na fase 2 com vazão afluente de 1,6 m3/h e TDH de 11,8 h. O TDH médio na câmara de entrada do RAH foi de 2,7h e de 9,1h na câmara ascendente de manto de lodo. Os reatores piloto apresentaram capacidade de tratamento para suportar os choques de carga orgânica devido à variabilidade do esgoto bruto. A eficiência de remoção de matéria orgânica em termos de DQO foi de 66% e 59% para o UASB e RAH, a eficiência de remoção de SST atingiu valores de 65% e 63% respectivamente, aliás, o reator UASB apresentando valores superiores de desempenho operacional em quase todos os parâmetros de monitoramento dos processos de digestão anaeróbia os valores médios dos parâmetros DQO, DBO5, SST e SSV não tiveram diferenças estatisticamente significativas. O pH para os sistemas piloto esteve na faixa ideal para o desenvolvimento dos microorganismos anaeróbios (6,5 a 7,5) e a alcalinidade e os AGVs demonstraram estabilidade dos reatores, corroborados pela estabilidade do pH no interior de cada sistema. A velocidade ascensional na câmara de manta de lodo (0,45 m/h) não causou efeito adverso na qualidade do efluente em termos de SST, com valor médio de 39,8 mg/L, na mesma faixa observada no reator UASB (41,6 mg/L). Foi observada menor concentração de biomassa no reator RAH (distribuição vertical) comparativamente ao reator UASB segundo os perfis verticais de lodo, entretanto, este fato não comprometeu a sua eficiência de remoção de matéria orgânica e dos sólidos suspensos. A retirada do lodo de excesso dos sistemas piloto foi baseada na relação STV/ST para estimar a fração de biomassa, a freqüência do descarte foi de 1 vez por mês retirando o equivalente aos 10% do volume de lodo das tomadas que apresentaram uma fração de STV menor que 50%. É viável a implementação da turbidez como parâmetro de detenção de perdida de lodo no efluente dos reatores anaeróbios devido a que a turbidez e os SST apresentaram uma relação direta o que ajudaria a simplificar mecanismos de controle operacional para a toma de decisões nas estações de tratamento, sendo que a turbidez é um parâmetro de leitura rápida “in situ” e poderia advertir do excesso de lodo ao interior do reator. Foram empregados diferentes métodos titrimétricos para a determinação dos AGVs no efluente do RAH e UASB. Após análise dos resultados, recomenda-se o uso do método de KAPP e RIPLEY para a determinação dos AGVs pela simplicidade metodológica e pela rapidez na obtenção do resultado. / This study aimed to evaluate the performance and operational stability of a new configuration of a hybrid anaerobic reactor (RAH) combining an anaerobic filter downflow followed by a sludge blanket chamber upflow compared to UASB in same operating conditions applied to sewage treatment. The evaluation of organic matter removal and removal efficiency of suspended solids in the pilot reactor was by testing COD, BOD, total suspended solids (TSS) and volatile suspended solids (VSS). The RAH and UASB reactors were operated for a period of 240 days in the first phase and 150 days in phase 2 with the inlet flow of 1,6 m3/h and 11,8h TDH. The average TDH RAH in the inlet chamber was 2.7h and 9,1h in ascending chamber sludge blanket. Pilot reactors showed treatment capacity to support organic shock loads due to the variability of raw sewage. The organic matter removal efficiency in terms of COD was 66% and 59% for the UASB and RAH, the TSS removal efficiency reached values of 65% and 63% respectively, by the way, the UASB presenting performance values higher operational in almost all parameters of the monitoring anaerobic digestion processes mean values of the parameters COD, BOD5, TSS and VSS were not statistically significant differences. The pH for the pilot systems was in the ideal range for the development of anaerobic microorganisms (6,5 to 7,5) and alkalinity and Volatile Fatty Acids (VFA) demonstrated stability of reactors corroborated by the pH stability within each system. The upflow velocity in the sludge blanket chamber (0,45 m/h) caused no adverse effect on effluent quality in terms of TSS, with an average of 39,8 mg/L, in the same range observed in UASB (41,6 mg/L). It was observed lower concentration of biomass in the reactor RAH (vertical distribution) compared to UASB second vertical sludge profiles, however, this fact did not compromise its removal efficiency of organic matter and suspended solids. The withdrawal of the pilot systems excess sludge was based on TSV/TS ratio to estimate the biomass fraction, the frequency of disposal was 1 once a month by removing the equivalent of 10% of the outlets sludge volume that had a fraction of TSV less than 50%. The implementation of turbidity as sludge lost detention parameter is viable in the effluent of the anaerobic reactors due to turbidity and TSS showed a direct relationship which would help simplify operational control mechanisms for decision making in treatment plants, wherein the turbidity is a quick read parameter "in situ" and could warn of excess sludge inside the reactor. Different titrimetric methods were employed for the determination of VFA and the effluent from the UASB and RAH. After analyzing the results, we recommend the use of KAPP and RIPLEY method for the determination of VFA by methodological simplicity and speed in obtaining results.