Este trabalho buscou avaliar o desempenho de um reator de leito fixo e escoamento ascendente, operado de modo contínuo, com argila expandida e espuma de poliuretano como suportes para imobilização da biomassa, no tratamento de água residuária proveniente de abatedouro de aves. Na primeira fase do trabalho, o reator foi operado em condição anaeróbia e anaeróbia-aeróbia, no tratamento da água residuária bruta proveniente do abatedouro. No entanto, sua eficiência na redução de matéria orgânica pode ser atribuída a processos de adsorção e não biodegradação, o que resultou em períodos de operação curtos devido à obstrução do leito reacional. Na segunda fase, o reator foi alimentado com água residuária coletada após o pré-tratamento por sistema de flotação por ar dissolvido (FAD), realizado no próprio abatedouro, com intuito de minimizar a carga lipídica e a concentração de sólidos na entrada do reator. Assim, a vida útil do reator foi prolongada, com eficiências médias de remoção de DQO e nitrogênio total de 90 \'+ OU -\' 4% e 44 \'+ OU -\' 10%, respectivamente, e eficiência de desnitrificação de 89 \'+ OU -\' 6%, para o reator anaeróbio-aeróbio com razão de recirculação do efluente líquido igual a 1,5. Assim, o reator demonstrou bom desempenho na remoção simultânea de matéria orgânica e nutrientes, desde que seja precedido por sistema de pré-tratamento para remoção parcial de sólidos e óleos e graxas. / This work aimed to evaluate the performance of an up-flow fixed bed reactor, operated in a continuous way, using expanded clay and polyurethane foam as supports for immobilization of the biomass, for treatment of poultry slaughterhouse wastewater. Initially, the reactor was operated under anaerobic and anaerobic-aerobic conditions for the treatment of poultry slaughterhouse wastewater. However, the COD removal efficiency was related to adsorption and not to biodegradation processes. In this way, the operational periods were short due to solids retention in the system. In the second part of this work, trying to reduce the lipids and solids concentration in the reactor\'s affluent, the wastewater was collected after a pre-treatment installed at the poultry slaughterhouse. This pre-treatment was constituted of a dissolved air flotation (DAF) system. As a result, the operational longevity of the reactor was increased and the COD and total nitrogen removal efficiencies were 90 \'+ OU -\' 4% and 44 \'+ OU -\' 10% respectively, and the denitrification efficiency was 89 \'+ OU -\' 6% under an anaerobic-aerobic condition with a recycle ratio of the liquid phase of 1,5. Therefore, the reactor showed a good performance in reducing the organic matter and removing nutrients simultaneously, since it is preceded by a pre-treatment for the partial removal of solids, oils and grease.

Este projeto estudou a influência do substrato na dinâmica do processo de formação do biofilme em espuma de poliuretano, em reatores anaeróbios horizontais de leito fixo diferenciais, alimentados com extrato de carne, glicose, amido, lipídeos e esgoto sanitário sintético. O estudo constitui em acompanhar a colonização das matrizes de espuma, ao longo do tempo, em relação a quantidade de biomassa aculumada, polímero extracelular produzido e características morfológicas das células presentes nos suportes retirados dos reatores diferenciais. O uso dessas técnicas permitiu o melhor entendimento do processo de aderência, além da verificação da composição morfológica e a estrutura do biofilme aderido ao suporte, possibilitando correlacionar a ocorrência de determinadas morfologias com cada etapa da colonização observada. A influência do substrato na dinâmica de aderência foi verificada devido aos diferentes padrões de colonização encontrados. Foi observada uma grande variabilidade morfológica em relação ao substrato utilizado. Todavia, uma grande ocorrência de organismos semelhantes a Methanosaeta sp foi verificada em todos os ciclos, principalmente em relação a Methanosarcina sp. O fenômeno de excreção de polímeros pareceu ser de fundamental importância no processo de colonização de matrizes de poliuretano, estando vinculado provavelmente à fixação das células aos suportes. A produção de polímeros, na etapa de aderência inicial, apresentou comportamento diferenciado para cada substrato utilizado. De acordo com os modelos propostos e as análises de microscopia eletrônica de varredura, verificou-se que os polímeros extracelulares podem estar ligados ao entupimento de reatores de leito fixo, permanecendo no interior dos suportes e nos interstícios do leito, causando problemas operacionais. As rápidas partidas observadas em trabalhos utilizando reatores de leito fixo e espuma de poliuretano como suporte podem estar vinculadas à rápida aderência dos organismos em todos os ciclos estudados. / This work focused on the influence of the type of substrate on the process of biofilm formation onto polyurethane foam matrices, in differential horizontal anaerobic immobilized sludge reactors, fed with meat extract (protein), glucose, starch, lipid and synthetic domestic wastewater. It consisted of accompanying the colonization of foam matrices with time, regarding to biomass amount, extracellular polymers produced and the morphological characteristics of the cells present on the supports packed in the differential reactors. These techniques permitted a better understanding of the adhesion process, besides a verification of the morphological composition and the structure of the biofilm attached to the support, making possible a correlation of the particular morphologies occurrences with each colonization step. The influence of the substrate was verified due to the different colonization patterns found. It was observed considerable morphological variety depending on the substrate utilized. However, the presence of Methanosaeta sp.- like organisms was often verified in all cycles, specially Methanosarcina sp. The excretion of polymers seemed to be crucial in the colonization process of the polyurethane matrices, being probably related to the cell fixation on the support. The polymeric production, in the inital adhesion step, showed a particular behavior for each substrate employed. According to the proposed models and the scanning electronic microscopy analysis, it was verified that the extracellular polymers can be related to the clogging of the fixed-bed reactors, as it keeps into the supports and the bed interstices, causing operational problems. The fast start-ups observed in works using fixed-bed reactors and polyurethane foam as support can be related to the fast cell adhesion during all cycles studied.

O presente trabalho teve como objetivo avaliar a influência da intensidade de agitação e tipo de impelidor, bem como a estratégia de alimentação, em reator anaeróbio operado em batelada seqüencial, escala piloto (\'da ordem de\' 1 \'M POT.3\'), com agitação mecânica e em duas configurações: uma com biomassa granulada (ASBR) e outra com biomassa imobilizada em suporte inerte de espuma de poliuretano (ASBBR), aplicadas ao tratamento de esgoto doméstico. No estudo da intensidade de agitação e tipo de impelidor, para cada reator foram avaliadas três tipos de impelidores (turbina de pás planas, turbina de pás inclinadas 45º e hélice) associados a duas intensidades de agitação (40 e 80 rpm), obtendo uma combinação de 6 (seis) condições experimentais. A combinação da intensidade de agitação e tipo de impelidor que apresentou melhor desempenho no processo foi utilizado no estudo da estratégia de alimentação, na qual foi avaliada as seguintes condições: batelada alimentada durante 25%, 50%, 75% e 100% do ciclo. Os resultados obtidos permitiram concluir que: no ASBBR o aumento da intensidade de agitação de 40 rpm para 80 rpm permitiu uma melhoria nos fluxos de transferência de massa e, portanto, aumentou a velocidade de consumo de substrato; no ASBR o aumento da intensidade de agitação de 40 rpm para 80 rpm proporcinou uma desestabilização do sistema, provavelmente por causa da ruptura dos grânulos provocada pela maior agitação; os sistemas operados com impelidor do tipo hélice apresentaram vantagens, tais como: melhor eficiência de remoção de sólidos, maior valor da constante cinética de primeira ordem (melhor fluxo de transferência de massa e conseqüentemente maior consumo de substrato) e maior produção de alcalinidade, ou seja, maior estabilidade para o sistema; tanto o sistema ASBR como o ASBBR quando operados nas condições de batelada típica, batelada alimentada durante 50% e 75% do ciclo apresentaram melhores desempenhos no processo de tratamento, mostrando a flexibilidade operacional dos reatores anaeróbios operados em bateladas seqüenciais; e comparando os sistemas ASBBR e ASBR verificou-se que estes apresentaram comportamentos similares em todas as condições de operação de batelada alimentada avaliadas, não sendo possível, estatisticamente apontar um sistema com melhor desempenho. / The objective of this work was to assess the effect of the stirring speed, the type of impeller and the feed strategy in a mechanically stirred pilot-scale (\'da ordem de\' 1 \'M POT.3\') anaerobic sequencing batch reactor to two configurations: a containing granulated biomass (ASBR) and the other containing immobilized biomass (AnSBBR). Domestic wastewater was treated in 8-h cycles. Three impeller types (turbine with six-flat blades, turbine with six 45º-inclined blades and helix with three blades) were assessed at two different stirring speeds (40 and 80 rpm), totaling six experimental conditions. The stirring speed and the impeller that resulted in the best combination was used in work of the feed strategy . The reactors were operated at room temperature at four different feed strategies (fed batch during 25%, 50% and 75% of the cycle, and conventional fed-batch). The results allowed conclude that: in the AnSBBR increasing the stirring speed from 40 rpm to 80 rpm showed to improve mass transfer, with consequent increase in substrate consumption; in the ASBR increasing the stirring speed from 40 rpm to 80 rpm showed desestabilization in system, because of the disruption caused in the granules witth greater agitation; operation with the helix impeller showed some advantages over the turbine impellers, such as: improved efficiency in solids removal, higher value of the first order kinetic constant and higher alkalinity production; both for the ASBR as for the ASBBR the best performance in wastewater treatment was obtained when the reactors were operated at conventional batch, fed-batch during 50% and 75% of the cycle; no significant difference in performance was observed among these three conditions. Despite poor performance of the conventional fed-batch and fed-batch during 25% of the cycle compared to the other conditions, both these conditions presented operational stability. Hence, the anaerobic sequencing batch reactors presented operational flexibility as far as feed strategy is concerned.

O projeto proposto teve como objetivo principal à avaliação do suporte no desempenho do reator anaeróbio operado em bateladas seqüenciais, com agitação, contendo biomassa imobilizada no tratamento de esgoto sanitário do Campus da USP em São Carlos. O estudo do desempenho global deste tipo de configuração de reator é extremamente vinculado com o suporte inerte o qual proporciona a aderência e crescimento da biomassa. Neste trabalho foram estudados três suportes inertes: espuma de poliuretano, carvão vegetal e polietileno de baixa densidade reciclado. A avaliação global foi realizada sob análise de três diferentes e dependentes variáveis locais: o desempenho do reator, a análise cinética e os exames microbiológicos. A espuma de poliuretano pôde ser considerada como sendo o material mais adequado para tal finalidade, apresentando maiores eficiências de redução da DQO de amostras brutas e filtradas de 61% e 71%, respectivamente, o melhor coeficiente cinético aparente de primeira ordem (k1T): 0,52 ± 0,04 h-1, e uma biomassa predominantemente caracterizada por arqueas metanogênicas e bactérias fototróficas. O carvão vegetal apresentou resultados intermediários, no entanto não satisfatórios, na redução de DQO e ainda uma biomassa não compatível com a esperada para um tratamento anaeróbio biológico. Ou seja, pode-se inferir que neste tipo de suporte houve seleção de biomassa inadequada para se tratar esgoto sanitário. Por fim, o polietileno apresentou os resultados menos expressivos, porém, dada sua caracterização microbiológica constatou-se ser um suporte com potencial para o tratamento de esgoto sanitário quando nele for aplicado um tratamento prévio, objetivando melhores características físico-químicos, como rugosidade, por exemplo. / The main objective of this study was the evaluation of bed materials on the performance of an anaerobic sequencing batch biofilm reactor, with agitation, treating sewage wastewater of the Campus of University of São Paulo in São Carlos. The study of the global performance of this reactor configuration is extremely linked with the bed material which provides adhesive and growing conditions. In this work three different bed materials were investigated : polyurethane foam, vegetal carbon and recycled low-density polyethylene. The global evaluation was performed under analysis of three different and dependant local variables: the reactor performance, kinetic analyses and microbial exams. The polyurethane foam was considered the most suitable material. Using the polyurethane foam as a bed material were obtained the highest COD removal efficiencies, 61% and 71% corresponding to total and filtered samples, respectively. In Addition, the polyurethane foam showed the best modified first-order kinetic model (k1T) of 0,52 ± 0,04 h-1, and a biomass predominately characterized by microorganisms resembling to methanogenic archea and phototrophic bacteria . The vegetal carbon showed intermediate results, which were not satisfactory enough, in the COD removal. Also, the vegetal carbon showed an incompatible biomass in accordance with the expected anaerobic biomass. In others words, it can be inferred that, using this type of material, there was a selective biomass process in which an inappropriate biomass was selected. Finally, the recycled low-density polyethylene showed the less expressive results. However, because of its microbial characterization, it can be concluded that the polyethylene material has potential to be applied as a bed material in the sewage wastewater treatment. Nevertheless, a previous treatment physics-chemical must be applied in this kind of bed material to improve the physics-chemical properties such as porosity and surface area.

Dentro do escopo de estudo de aspectos fundamentais e tecnológicos, este trabalho foi direcionado à avaliação do comportamento da estabilidade e da eficiência de um reator anaeróbio contendo biomassa imobilizada em espuma de poliuretano para o tratamento de água residuária sintética com concentração de aproximadamente 500 mgDQO/L. O reator, de 6 litros de volume nominal, mantido à temperatura de 30 +/- 1ºC e sob a agitação mecânica de 500 rpm, foi operado em bateladas seqüenciais com a renovação de apenas parte do volume de água residuária tratada por ciclo, ou seja, a descarga do reator não foi completa, mas apenas parcial. Neste contexto, a principal característica operacional investigada foi a relação entre o volume de água residuária afluente, o qual era alimentado a cada novo ciclo de operação, e o volume de meio contido no reator, o qual não era descarregado no ciclo anterior, mantendo-se a mesma carga orgânica volumétrica. Desta forma, foi possível verificar a flexibilidade operacional do sistema com relação ao volume de água residuária tratada a cada ciclo e o tempo de duração do ciclo para uma mesma carga orgânica. Os resultados indicaram que o reator pôde operar com diferentes relações entre o volume alimentado e o volume de meio reacional sem diminuição significativa do seu desempenho, permitindo-se, assim, maior flexibilidade operacional. Para as condições em que o volume renovado foi maior ou igual a 50% do volume de meio do reator (ciclos de 4 a 8 horas), as eficiências de remoção de matéria orgânica filtrada e não filtrada mantiveram-se no patamar de 84 e 79%, respectivamente, enquanto que nas demais condições de maior diluição inicial do afluente (ciclos de 1 e 2 horas), essas eficiências foram ligeiramente menores, em torno de 80 e 74%, respectivamente. A alta velocidade de formação de material viscoso de aparência polimérica, nas condições de maior diluição inicial do afluente, dificultou a manutenção do volume de meio reacional do reator constante. / An assessment was performed on the stability and efficiency of an anaerobic reactor containing biomass immobilized on polyurethane foam for the treatment of synthetic wastewater with organic loading of approximately 500 mgCOD/L. The reactor had a total volume of 6 L. Batch operation took place at a temperature of 30 +/- 1ºC and an agitation rate of 500 rpm. During operation only part of the treated wastewater was renewed per cycle, i.e., reactor discharge was not complete, but partial. The main operation characteristic investigated was the ratio between influent wastewater volume, which was fed at each cycle, and the volume of medium contained in the reactor, which had not been discharged in the previous cycle, maintaining the same volumetric organic load. This way, it was possible to verify operation flexibility of the system regarding treated wastewater volume in each cycle as well as cycle time duration for the same volumetric organic loading. Results indicated that the reactor was able to operate at different ratios between feed and reaction medium volume without significant loss in performance, allowing, hence, greater operation flexibility. For conditions at which renewed volume was greater than or equal to 50% of the reactor medium (cycles from 4 to 8 hours), filtered and non-filtered organic matter removal efficiencies were about 84 and 79%, respectively, whereas at conditions of higher initial influent dilution (cycles from 1 to 2 hours) these efficiencies were slightly lower, about 80 and 74%, respectively. The high formation rate of viscous polymer-like material probably of microbiological origin, at the conditions of higher initial influent dilution hindered maintenance of constant reactor medium volume.

Cette étude vise à produire deux enzymes amylolytiques (α-amylase et pullulanase) thermostables par des levures contaminant le blé récolté dans des zones semi arides et arides (Biskra - Sahara, Sud Algérien) et capables d’hydrolyser à la fois les liaisons α-1-4 et α-1-6 de polysaccharides comme l’amidon et le pullulane, molécules d’intérêt industriel. Après isolement et caractérisation de colonies levuriennes, la méthode de plate-test-agar permet d’isoler des souches amylolytiques et de montrer que la souche L7 est la plus performante dans la production enzymatique parmi une douzaine de souches de levures productrices d’α-amylase et de pullulanase thermostables. L’identification des souches, basée sur les caractères morphologiques, les tests biochimiques et la biologie moléculaire a permis de répartir la population comme suit : 50% Clavispora lusitaniae (forme anamorph Candida lusitaniae), 25%, Pichia guilliermondii, 8% Pichia carribbicca, 8% Meyerozyma guilliermondii et 8% Rhodotorula rubra. Par sa richesse en amidon, le biotope du blé est favorable à la survie des levures amylolytiques. La majorité de ces souches dont la souche L7 est productrice de pseudo ou vrai mycélium et est tolérante à certains paramètres comme la température, la salinité, les stress osmotique et éthanolique. La souche de levure L7, Clavispora lusitaniae ABS7, semble être la plus performante dans la production d’enzymes thermostables. Son identification moléculaire a montré deux bandes avec l’endonucléase HAE III alors que les autres souches de la même espèce de Clavispora lusitaniae (L5, L9, L10, L11 et L12) présentent une seule bande. En conditions optimales (agitation 136,56 rpm, température 54,14°C, amidon 2,66g/l, extrait de levure 0,365g/l, sels 8, 75ml/l et oligo-éléments 4,3ml/l en erlenmeyers de 250 ml), la production maximale atteint les valeurs suivantes : 13456,36±300 UI pour l’ α-amylase et 12611, 6±154 UI pour la pullulanase. Ces performances sont en accord étroit avec la prédiction du modèle statistique évaluée à 13231UI pour l’α-amylase et 12825,5 UI pour la pullulanase. La production optimisée a pratiquement doublé par rapport à la production avant l’optimisation (6639,16 UI pour l’α-amylase et 6308,5 UI pour la pullulanase). En conditions optimales et en fermenteur de 2 L, la production maximale pour les deux enzymes de la levure Clavispora lusitaniaeABS7 est obtenue au bout de 28 h avec un optimum de croissance obtenu à 40 heures. La production des deux enzymes n’est donc pas associée à la croissance. La production maximale des deux enzymes s’effectue à pH 8. pic protéique. L’élution sur DEAE-cellulose confirme la présence des deux activités dans la même fraction. Les deux enzymes sont donc présentes sur la même molécule. L’α-amylase et la pullulanase sont purifiées avec un taux de purification de 50,5 et 44,6 respectivement et des rendements respectifs de 23,9% et 21,1%. L’extrait purifié montre une seule bande sur le gel de SDS-PAGE avec un poids moléculaire estimé à 75KDa et une activité amylolytique contenant à la fois les activités α-amylasique (indépendante de Ca2+) et pullulanasique (une métalloenzyme à calcium). La souche de la levure Clavispora lusitaniae ABS7 possède donc une enzyme amylolytique avec deux sites actifs. La CCM révèle une enzyme qui hydrolyse l’amidon en maltose et glucose et le pullulane en maltotriose, maltose et glucose, ce qui montre que l’enzyme est saccharifiante et correspond à une pullulanase de type II (amylopullulase). L’optimisation de l’immobilisation de l’enzyme a permis l’amélioration de l’activité: α-amylasique à 4907,75 UI (rendement 72,3 %) et celle de la pullulanase à 4491,83 UI (rendement 70,1%) avec un pH optimum de 8,5. Il ressort de notre étude que l’amylopullulanase type II libre de Clavispora lusitaniae ABS7 est thermostable puisqu’elle résiste à un traitement thermique de 75°C pendant 3 heures d’incubation et conserve 88% de son activité initiale. / This study aims to produce two amylolytic enzymes (α-amylase and pullulanase) by thermostable wheat contaminant yeast harvested in semi arid and arid zones (Biskra, Sahara, Algeria SUD) and capable of hydrolyzing both the α links 1-4 and 1-6 of polysaccharides such as starch and pullulan, molecules of industrial interest. After isolation and characterization of levuriennes colonies, the test method of agar-plate allows to isolate amylolytic strains and show, that the L7 strain is the most effective, in the enzymatic production of the 12 yeast strains producing α-amylase and pullulanase the thermostable. The identification of strains, based on morphological, biochemical tests and molecular biology has helped spread the population as follows: 50% Clavispora lusitaniae (anamorph form Candida lusitaniae), 25%, Pichia guilliermondii, 8% carribbicca Pichia, 8% Meyerozyma guilliermondii and 8% Rhodotorula rubra. By its high starch, the wheat biotope is favorable to the survival of amylolytic yeasts. Most of these strains, including the strain L7, is producer, pseudo or true mycelium and is tolerant to certain parameters such as temperature, salinity, osmotic stress and ethanolic stress. The yeast strain Clavispora lusitaniae ABS7 (L7) seems to be the most efficient in the production of thermostable enzymes. Its molecular identification showed two bands with the endonuclease HAE III while other strains of the same species Clavispora lusitaniae (L5, L9, L10, L11 and L12) have a single band. In optimal conditions (agitation 136.56 rpm, temperature 54.14 ° C, starch 2,66g / l, yeast extract 0,365g / l, salts 8 75ml / l and trace elements 4,3ml / liter Erlenmeyer flasks into 250 ml), the maximum production reached: 13456.36 ± 300 IU for the α-amylase and 12611, 6 ± 154 IU for pullulanase. This performance is in close agreement with the prediction of the statistical model 13231UI evaluated for α-amylase and 12825.5 IU for pullulanase. The optimized production almost doubled compared to production before optimization (6639.16 IU for the α-amylase and pullulanase for 6308.5 IU). In optimal conditions, and 2 L fermenter, the maximum production for the two enzymes of Clavispora lusitaniae ABS7 obtained after 28 hours, with an optimum of growth obtained at 40 hours. The production of both enzymes is thus not associated with growth. The maximum production of both enzymes is obtained at pH 8. The kinetics are characterized by an increase in carbohydrate and a substance spooning the wall of the fermenter, probably an exo-polysaccharide. The chromatographic profile on Sephacryl S200 reveals two α-amylase and pullulanase activities eluted along with the protein peak. Elution DEAE cellulose confirms the presence of both activities in the same fraction. Both enzymes are present on the same molecule. The α-amylase and pullulanase were purified with a purification rate of 50.45 and 44.59 respectively and respective yields of 23.88% and 21.11%. The purified enzyme showed a single band on SDS-PAGE gel with a molecular weight estimated at 75 KDa and an amylolytic activity containing both the α-amylase activities (independent of Ca2+) and pullulanase (a calcium metalloenzyme). The strain of the yeast Clavispora lusitaniae ABS7 therefore has an amylolytic enzyme with two active sites. TLC reveals an enzyme which hydrolyzes starch into maltose and glucose and pullulan into maltotriose, maltose and glucose, which shows that the saccharifying enzyme, and corresponds to a pullulanase type II (amylopullulase). The optimization of the immobilization of the enzyme enabled the improvement of the activity: α-amylase to 4907.75 IU (yield 72.3%) and pullulanase to 4491.83 IU (yield 70, 1%) with a pH optimum of 8.5. It appears from our study that amylopullulanase type II free is thermostable to heat treatment of 75 ° C for 3 hours of incubation, and retains 88% of its original activity.

Ce travail a pour objectif la conception et l'étude d'un réacteur enzymatique à membrane (REM) en vue de la dégradation de composés phénoliques. Pour cela, des membranes actives ont été préparées par greffage covalent de la laccase de Trametesversicolor à la surface d'une membrane céramique Après avoir mis en évidence les potentialités du réacteur vis-à-vis de la dégradation du 2,6-diméthoxyphénol (DMP) choisi comme substrat modèle, l'impact de paramètres opératoires (débit d'alimentation, concentration en substrats) sur les performances du REM a été étudié. Pour résoudre des problèmes d'instabilité, différentes étapes du protocole de fabrication des membranes actives ont été revues. Puis les paramètres opératoires (pH et température) ont été étudiés afin d'optimiser les conditions de mise en œuvre du REM. Il a été établi que les performances épuratoires étaient maximales à pH acide (pH 4) mais restaient stables sur une large plage de températures (15 à 40°C). Enfin la dernière partie de la thèse a été consacrée à l'étude du colmatage et à la recherche de différentes stratégies visant à limiter l'impact de ce dernier sur les performances et la stabilité du REM. / This work was devoted to the design and the study of an enzymatic membrane reactor (EMR) for the degradation of phenolic compounds. For this, active membranes were synthesized by covalent grafting of laccase from Trametes versicolor on the surface of a porous ceramic membrane.The 2,6-dimethoxyphenol (DMP) was chosen as model substrate and the effect of operating parameters (feed flow rate, substrate concentration) on the performance of the EMR have been studied.Different stages of the active membrane preparation process were reviewed in order to improve the system stability. Moreover operating parameters (pH and temperature) were studied in order to optimize the performance of the EMR. It was shown that the depletion efficiency was maximal at relatively low pH (pH 4) but remained stable over a wide temperature range (15 to 40 ° C).Finally the last part of the work was devoted to the developmentof different strategies to limit the impact of fouling on the performance and stability of the EMR.

A utilização de reatores anaeróbios de fluxo ascendente com manta de lodo no tratamento de águas residuárias tem tido grande aceitação dos projetistas e pesquisadores pelas vantagens que esse sistema apresenta sobre os demais (aeróbios e anaeróbios). Uma das principais limitações do processo relaciona-se com o período de partida, normalmente lento e de difícil controle. Esse trabalho apresenta os resultados de pesquisas em laboratório sobre a partida de dois reatores de manta de lodo, um deles inoculado com lodo anaeróbio imobilizado em matrizes de gel agar e o outro com lodo livre. A imobilização de microrganismos em gel surge como uma nova possibilidade de retenção da população microbiana no reator e consequentemente, esse trabalho objetiva também verificar a viabilidade do uso de lodo imobilizado em qel agar no estudo sobre fatores que afetam o desempenho de reatores com manta de lodo. Os dois reatores anaeróbios utilizados neste trabalho eram idênticos e possuiam volume de 10 L cada um. Utilizou-se substrato sintético para que se obtivesse alimento com características estáveis durante a maior parte do tempo necessário à partida dos reatores, preparados de maneira a se obter DQO de aproximadamente 2000 mg/L. Os reatores estiveram em operação durante 179 dias e foram submetidos a cargas orgânicas volumétricas que variaram de 0,47 a 12,83 KgDQO.m-3.d-1 e a cargas hidráulicas volumétricas que variaram de 0,23 a 6,0 m3.m-3.d-1. As principais análises realizadas foram DQO, alcalinidade, ácidos voláteis, pH, temperatura e altura do leito de lodo. A eficiência de remoção de DQO esteve em geral entre 70 e 80%. Em vista dos resultados obtidos, conclue-se que a utilização de lodo anaeróbio imobilizado em matrizes de gel agar pode ser útil no estudo de partida de reatores anaeróbios com manta de lodo. Sugere-se a continuidade dos estudos, utilizando-se as técnicas descritas no trabalho, na ampliação do conhecimento dos processos anaeróbios neste tipo de reator. / The use of Upflow Anaerobic Sludge Blanket (UASB) Reactor for wastewater treatment has been well accepted by designers and researchers due to the advantages shown over athers reactors (aerobic and anaerobic). The reactor start-up remains as one of the most important limitations of the process. It is usualy slow and is difficult to control. This thesis shows the results of a laboratory research on the start-up of two UASB reactors. One of them was seeded with anaerobic sludge immobilized in gel agar and the other with free anaerobic sludge. The immobilization of microorganisms in gel represents a new possibility of biomass retention within the reactors. Consequently this research also aims to check the viability of using immobilized sludge in gel agar in the study of factors affecting UASB reactors performance. Both the UASB reactors used in this research were exatly alike and had a volume 10 L each. Synthetic substrate was used to provide, consistent feed during the most of the start-up period. The substrate was prepared to contain about 2000 mg/L COD. The reactors were aperated for 179 days and were submitted to organic loading rates which varied from 0,47 to 12,83 KgCOD.m-3.d-1 and to hydraulic loading rates which varied from 0,23 to 6,0 m3.m-3.d-1. The main physical-chemical parameters measured were COD, alkalinity, volatite fatly acids, pH, temperature and sludge bed height. The COD removal efficiency was between 70 and 80%. This research comproved the feasibility of sludge immobilized in gel agar for UASB reactor start-up. It is suggested to continue the studies to achieve the best conditions for the use of this technique.

The main purpose of the master thesis project is to investigate the influence of selected serine protease inhibitors (SPI) on the catalytic action of the serine proteases chymotrypsin and trypsin, in a conjugated and non-conjugated state. The inhibitors included for this study were extracted from Solanum tuberosum, i.e.common potato. The purification method included in this study consist of crude extraction by mixer, followed by a salt-out procedure with ammonium sulphate. Further purification steps were cation exchange chromatography and, finally, gel filtration to obtain SPI of high purity. The purified sample was then characterized by SDS-page and kinetic activity measurement of trypsin and chymotrypsin action on synthetic substrate derivate, N-Benzoyl-DL-arginine-4-nitroanilide hydrochloride (BAPA) and N-Succinyl-L-phenylalanine-p-nitroaniline (SFpNA) respectively. The characterization showed inhibitory inactivation of both pancreatic proteases. This would indicate successful extraction of SPI. To investigate inhibitory action in a conjugated state, either enzyme or inhibitor was immobilized onto aluminium oxide membranes. Then two different experimental setups were tested, called experiment 1 and 2. In experiment 1, the inhibitor was immobilized and the interaction was monitored from a retention shift of enzyme flow-through compared to a blank column, using detection at 280 nm of the enzyme. In experiment 2 the enzyme was instead immobilized and a mixture of inhibitor and substrate was circulated with monitoring of the catalytic activity. The main goal was thus to measure the effects on the kinetics in the conjugated state compared to enzyme and inhibitor in the free state. The result from both experiment 1 and 2 did not yield consistent and reliable result so the discussed method should be regarded as preliminary results. The study also includes investigation of inhibitor-enzyme interaction as revealed by molecular mass data to determine complex formation. This part was conducted with static light scattering analysis to determine the stoichiometry for the interaction between pancreas proteases and the inhibitor. Results from light scattering showed promising indication of many-to-one interaction between enzyme and inhibitor, which have been seen by previous studies. It should be considered a preliminary result as complex formation does not exclude aggregation of enzymes or inhibitor in the solution.

O projeto consistiu na concepção e avaliação do desempenho de um sistema anaeróbio-aeróbio para tratamento de esgoto sanitário. O leito do reator foi dividido em quatro compartimentos de igual volume. Foi usada espuma de poliuretano para imobilização da biomassa e, para facilitar a partida do reator, esse material suporte foi previamente inoculado. O projeto teve três fases distintas: na primeira etapa, foi observada a importância da concentração de biomassa anaeróbia no desempenho de reator anaeróbio a princípio operado com metade do leito reacional com espuma e, a seguir, com o leito todo preenchido com espuma. Na segunda etapa, foram testados diferentes tempos de detenção hidráulica no reator que operou apenas em condições anaeróbias. Na última etapa foi operado o reator combinado anaeróbio-aeróbio. Ficou constatada a importância da concentração de microrganismos no desempenho do reator anaeróbio, pois com o aumento da quantidade de espuma, o reator atingiu resultados melhores e maior estabilidade operacional. Foi comprovada a influência do tempo de detenção hidráulica (TDH) no desempenho do reator. Tempos de detenção muito altos acarretam problemas de transferência de massa líquido-sólido, e baixos TDH dificultam adequada ação dos microrganismos. O melhor resultado para o reator em operação exclusivamente anaeróbia foi para o TDH de 10 horas, no qual se conseguiu reduzir a DQO de amostra bruta de 389 +/- 70 mg/L para 137 +/- 16 mg/L, em média. Para o reator operado anaeróbio-aeróbio a DQO de amostra bruta decresceu de 259 +/- 69 mg/L para 93 +/- 31 mg/L, em média. A comparação de todos os resultados obtidos, evidenciou a importância do pós-tratamento aeróbio na remoção de parcela de matéria orgânica não removida em tratamento unicamente anaeróbio. / The project consisted in the design and performance evaluation of an anaerobic-aerobic system for wastewater treatment. Polyurethane foam was used for biomass immobilization and, to smooth the reactor start-up, this supporting material was previously inoculated. The project was divided in three distinct phases. In the first one, the importance of the anaerobic biomass concentration was observed in the performance of the anaerobic reactor in a way to operate the reactor primarily with half of the reaction bed filled with foam and subsequently operate it with the bed completely filled with foam. In the second phase, different times of hydraulic retention were tested with the reactor operating exclusively in anerobic conditions. In the third and last phase, an anaerobic-aerobic combined reactor was operated. It was possible to confirm the importance of microorganism concentration in the performance of the anaerobic reactor, since the increase in the amount of foam allowed the reactor to reach better results and greater operational stability. The influence of the hydraulic retention time in the reactor performance was also proved. Very high retention times cause problems in the liquid-solid mass transference, while low retention times do not allow an adequate action of microorganisms. The best result for the reactor with an exclusive anaerobic operation was the 10 hour retention time, when it was possible to reduce the COD of a 389 +/- 70 mg/L gross sample to a 137 +/- 16 mg/L in average. On the other hand, for the anaerobic-aerobic operating reactor, the COD of a gross sample dropped from 259 +/- 69 mg/ L to 93 +/- 31 mg/L in average. Finally, comparing all the obtained results, it was possible to verify the importance of the anaerobic post treatment in the removal of a part of the organic matter not removed in an exclusively anaerobic treatment.