O objetivo desse estudo foi averiguar a possibilidade de se estimar os componentes anaeróbios do déficit máximo acumulado de oxigênio (MAOD) utilizando apenas o consumo de oxigênio (VO2). Adicionalmente, foi proposto um método alternativo que não necessita utilizar a extrapolação do VO2 do exercício submáximo para estimar a demanda de O2 para o exercício supramáximo. Para tanto, dez sujeitos fisicamente ativos foram submetidos aos seguintes testes: a) teste progressivo até a exaustão para determinar a carga correspondente ao consumo de oxigênio (WVO2max); b) seis testes de cargas constantes com intensidades abaixo da WVO2max (40-90% WVO2max); c) um teste a 110% da WVO2max; d) teste anaeróbio de Wingate. A fração do metabolismo anaeróbio alático foi estimada pela fase rápida do excesso do VO2 após o exercício (EPOCRÁPIDO), ao passo que a fração do metabolismo anaeróbio lático foi determinada tanto pela diferença entre MAOD e o EPOCRÁPIDO (MAODLA-1) como pelo acúmulo de lactato no sangue (MAODLA-2). O MAODMOD foi calculado pela somatória do EPOCRÁPIDO com MAODLA-2. Os principais resultados foram: a) as estimativas das contribuições do metabolismo anaeróbio lático não eram estatisticamente diferentes (MAODLA-1 = 2,40 ± 0,66 l; MAODLA-2 = 2,20 ± 0,4 l; p > 0,05); b) similarmente, os resultados do MAOD (3,03 ± 0,62 l) MAODMOD (2,84 ± 0,41) não eram significativamente diferentes (p > 0,05); c) foram detectadas correlações positivas entre MAODMOD, EPOCRÁPIDO, MAODLA-2 e os índices do teste de Wingate; d) os valores percentuais do EPOCRÁPIDO e do MAODLA-1 eram 22 ± 9% e 78 ± 9%, respectivamente. Esses resultados sugerem que os componentes anaeróbios do MAOD podem ser estimados satisfatoriamente utilizando somente o VO2 e que o MAOD pode ser determinado mediante MAODmod / The objective of this study was ascertain the possibility of estimate the anaerobic components of the maximal accumulated oxygen deficit (MAOD) used only oxygen uptake (VO2). In addition, was proposed an alternative method (MAODMOD) to determine the MAOD without using the extrapolation from submaximal VO2 to estimate the O2 demand of supramaximal exercise. Thus, ten subjects physically active performed the following tests: a) a maximal incremental exercise test for the measurement of the power output correspondent to maximal oxygen uptake (WVO2max); b) six submaximal tests with intensities bellow of the WVO2max (40-90% WVO2max); c) a supramaximal test at 110% WVO2max; d) Wingate anaerobic test. The fraction of the anaerobic alactic metabolism was calculated based on the fast component of excess post-exercise oxygen uptake (EPOCRÁPIDO), although the fraction of the anaerobic lactic metabolism was calculated based on both difference between MAOD and EPOCRÁPIDO (MAODLA-1) and changes in net blood lactate accumulation (MAODLA-2). MAODMOD was calculated by the sum of the EPOCRÁPIDO with the MAODLA-2. The main results were: a) non significant difference was found between the anaerobic lactic estimates (MAODLA-1 = 2,40 ± 0,66 l; MAODLA-2 = 2,20 ± 0,4 l; p > 0,05); b) similarly, there was no significant difference (p > 0,05) between MAOD (3,03 ± 0,62 l) and MAODMOD (2,84 ± 0,41); c) positive and significant correlation were detected between MAODMOD, EPOCRÁPIDO, MAODLA-2 and the indices of the Wingate anaerobic test (p < 0,05); d) the mean of the percentage values of the EPOCRÁPIDO and MAODLA-1 were 22 ± 9% e 78 ± 9%, respectively. These data suggest that the anaerobic components of the MAOD can be satisfactorily estimated using only the VO2 and that the MAOD can be determined using the MAODmod

O objetivo do presente trabalho foi realizar estudos acerca da degradação anaeróbia de proteínas no tocante à cinética de consumo de substrato e às rotas metabólicas presentes no processo. Inicialmente, realizou-se um estudo comparativo da degradação de efluentes sintéticos em reatores anaeróbios horizontais de leito fixo (RAHLFs) em escala de bancada. Nesse intuito, foi proposto um modelo cinética de primeira ordem de reações em série e paralelo, que permitiu a estimativa dos parâmetros cinéticos das várias etapas do processo estudado. As fontes de carbono nos efluentes sintéticos foram primeiramente compostas por compostos protéicos, tendo sido posteriormente adicionadas fontes de carboidratos e lipídeos à sua composição. Nesse intuito, utilizaram-se duas proteínas, caseína e soro albumina bovina (S.A.B.), e um hidrolisado protéico, peptona. Os resultados preditos pelo modelo cinético proposto apresentaram boa correlação com os dados experimentais e, com isso, foi possível o entendimento dos vários aspectos envolvidos na degradação de compostos protéicos em reatores que apresentam regime de escoamento predominantemente pistonado. A peptona foi mais rapidamente consumida, seguida pela caseína e finalmente pela S.A.B.. A presença de lipídeos nos efluentes sintéticos utilizados se constituiu no maior problema para o processo estudado. Análises de microscopia e biologia molecular mostraram que houve realocação da biomassa nos RAHLFs à medida que foram adicionadas fontes alternativas de carbono aos efluentes sintéticos à base de proteína. As morfologias encontradas foram comuns em todos os ensaios realizados, tendo predominado as semelhantes a Methanosaeta sp., bacilos fluorescentes e não fluorescentes, bacilos curvos e cocos não fluorescentes. Também foram visualizados três tipos de filamentos, espiroquetas e morfologias semelhantes a Methanosarcina sp. A segunda etapa do presente trabalho foi o estudo comparativo do processo de degradação de água residuária proveniente de abatedouro de aves em dois reatores um UASB e um RAHLF. O modelo cinético de primeira ordem se ajustou bem aos dados de decaimento de matéria orgânica em ambos os reatores. Dessa forma, foi possível concluir que no reator UASB, o mecanismo de adsorção do material colóide e solúvel nos grânulos parece ter regulado o processo, resultando em maior velocidade de remoção dessas frações de matéria orgânica. No RAHLF, o que parece ter regulado o processo foi a retenção de sólidos orgânicos, aumentando, dessa forma, a velocidade de remoção dessa fração de matéria orgânica. / This work reports on the anaerobic degradation of proteins from the standpoint of substrate consumption kinetics and metabolic routes. Initially, a comparative study of synthetic effluents in a bench scale horizontal-flow anaerobic immobilized biomass (HAIB) reactor was carried out. For this purpose, a kinetic model of irreversible first-order series-parallel reactions with two intermediate products was proposed. This allowed the estimative the kinetic parameters of several steps of the process. The carbon source in the synthetic effluents was composed primarily of casein, peptone and bovine serum albumin (B.S.A.). Then, sources of carbohydrate and lipid were added to the composition. The proposed model fitted the experimental protein degradation data well and allowed the best comprehension of the process. Thus it was possible to understand several aspects about the degradation of the protein substrates utilized in HAIB reactors. In relation to degradation rate, it was concluded that peptone was the most rapidly consumed, followed by casein, and finally BSA. Regarding substrate composition, the major problem for the studied process was the presence of lipids in the synthetics effluents. The results obtained from the microbiological analysis showed that relocation of biomass in the HAIBs reactors happened in correlation with the addition of alternative carbon sources to synthetic protein effluents. In all assays, common morphologies were found. These werepredominantly Methanosaeta sp. - like archaeas, fluorescent bacillus (three types), non-fluorescent bacillus (6 types), curved bacillus and non-fluorescent coccus. Three types of filaments, spirochetes and Methanosarcina sp. - like archaeas were also found. In a second phase, a comparative analysis of the degradation process of wastewater from a poultry slaughterhouse was carried out in two different reactors, one up flow anaerobic sludge blacked (UASB), and one HAIB. The first order kinetic model proposed fitted the experimental organic matter degradation data well in UASB and HAIB reactors treating residual water from avian abattoirs. In the UASB reactor, the adsorption mechanism of the granules for colloid and soluble material seems to regulate the process. This results in a higher rate of removal for these fractions of organic material. However, in the HAIB reactor, what seems to be controlling the process was the retention of organic solids, which increased the removal rate for this fraction of organic material.

Anaerobic digestion (AD) is becoming a widely adopted technology for conversion of organic waste and nutrient-rich fertiliser production due to its cost-effectiveness and sustainability. In this study, a batch experiment was conducted using five different types of food waste and cow dung (CD). No significant difference was observed among the four substrates that produced the highest methane (P<0.05). Based on the batch experiment results, two substrates were selected for semi-continuous digestion and the highest methane yield (67%) was obtained from co-digestion (CO). PCR-DGGE results revealed higher bacterial and archaeal diversity indices in CO as compared to mono-digestion of CD and mixed food waste. The high-throughput sequence analyses revealed that the Operational Taxonomic Units (OTUs) belonging to the phyla Bacteroidetes, followed by Firmicutes, Actinobacteria and Proteobacteria, were dominant in all treatments. The enhanced methane production in CO could be attributed to the neutral pH and partial shift of archaea from Methanosaeta to Methanosarcina. The digestate and fresh CD were screened for plant growth promoting bacteria (PGPB), nutrient and heavy metal content. The dung contained higher concentrations of heavy metals (P<0.05) and potential pathogens in comparison to the digestate. The use of digestate may, therefore, enhance soil fertility with minimal negative environmental effects. / School of Agriculture and Life Sciences / M. Sc. (Life Sciences)

Made available in DSpace on 2014-06-11T19:22:52Z (GMT). No. of bitstreams: 0 Previous issue date: 2004-05-17Bitstream added on 2014-06-13T19:49:12Z : No. of bitstreams: 1 afonso_m_me_rcla.pdf: 222236 bytes, checksum: 1a0edd0d0141d788c0b3436c47d1f119 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O limiar anaeróbio (LAN) tem sido muito utilizado como preditor da capacidade aeróbia na avaliação e prescrição do treinamento. Dentre os protocolos de determinação do LAN, o método do lactato mínimo (LM) parece ser um dos mais precisos, determinando de certa forma, individualmente os valores do LAN. Um outro modelo utilizado na avaliação do parâmetro aeróbio é o método não invasivo de determinação de potência crítica (PC), sendo que esse modelo possibilita também a quantificação das reservas anaeróbias intramusculares (CTA). Vários autores propõem determinações de PC por diferentes protocolos, dentre eles, os propostos por JENKINS & QUIGLEY em 1991 (PCJQ) e CHASSAIN em 1986 (PCCH) especialmente, são muito diferentes, no aspecto metodológico e repercutiram bastante no meio científico. O objetivo do presente estudo foi de comparar métodos invasivos e não invasivos de avaliação aeróbia e anaeróbia em atletas ciclistas, identificando a relação existente entre o LM e a PC (determinada por dois métodos diferentes) e, a relação dos parâmetros anaeróbios, através da potência média e da potência pico com a CTA. Foram voluntários desse estudo 9 ciclistas treinados do sexo masculino (24,11 1,62 anos; 67,28 4,37 kg e 175,78 5,89 cm) com experiência na modalidade. Os atletas foram submetidos a três protocolos experimentais: 1) Teste de LM, com teste de Wingate para induzir a acidose e posterior exercício progressivo; 2) Teste de PCJQ com quatro sessões de cargas exaustivas randômicas; 3) Teste de PCCH a partir da variação da freqüência cardíaca e do lactato através de dois esforços de 180 segundos com intervalo de pausa de 90 segundos entre os esforços, com intensidade de 85, 95, 105 e 115% do LM. Foram encontradas diferenças significativas entre as intensidades de LM (219,73 4,43 W), PCJQ nos modelos Pot- 1/tlim... . / The anaerobic threshold (AT) has been used as a good predictor of the aerobic capacity for physical evaluation and training prescription. Among the protocols of AT determination, the lactate minimal test (LMT) seems to be one of the most accurate for obtain individually the AT values. Another model used in the aerobic evaluation is the non-invasive method for determination of the critical power (CP), and this model also makes possible the quantification of the intramuscular anaerobic stores, which represents the anaerobic work capacity (AWC). Several authors proposed determinations of CP on different protocols, as those propored by, Jenkins & Quigley (1991 - CPJQ) and Chassain (1986 - CPCH), that in spite of the very different methodological aspects they are a reference in the scientific way. The purpose of the present study was to compare invasive and non-invasive protocols to evaluate the aerobic and anaerobic fitness (by two different methods) in cyclists. Also, the relationship of the mean and peak power obtained during an Wingate test to AWC identified by different methods was investigate. Nine male trained cyclists volunteered to the study (24.11 l 1.62 years; 67.28 l 4.37 kg and 175.78 l 5.89 cm). The athletes were submitted to three experimental protocols: 1) LMT, with initial lactic acidosis induced thought a Wingate test and a subsequent incremental exercise test; 2) test of PCJQ with four sessions of random exhaustive loads; 3) test of PCCH based on the variation of the heart rate and lactate along two bouts of efforts of 180 seconds with pause interval of 90 seconds between them, with intensity of 85, 95, 105 and 115% of LMT. Our results showed significant differences among the intensities of LMT (219.73 l 4.43 W), PCJQ in the models P-1/tlim (301.47 l 10.46 W) and Wlim - tlim (295.22 l 10.14 W) and, in PCCH for the methods of lactate variation (201.53 l 4.44 W) and... (Complete abstract, click electronic address below).

Orientador: Claudio Alexandre Gobatto / Banca: Eduardo Kokubun / Banca: Herbert Gustavo Simões / Resumo: O limiar anaeróbio (LAN) tem sido muito utilizado como preditor da capacidade aeróbia na avaliação e prescrição do treinamento. Dentre os protocolos de determinação do LAN, o método do lactato mínimo (LM) parece ser um dos mais precisos, determinando de certa forma, individualmente os valores do LAN. Um outro modelo utilizado na avaliação do parâmetro aeróbio é o método não invasivo de determinação de potência crítica (PC), sendo que esse modelo possibilita também a quantificação das reservas anaeróbias intramusculares (CTA). Vários autores propõem determinações de PC por diferentes protocolos, dentre eles, os propostos por JENKINS & QUIGLEY em 1991 (PCJQ) e CHASSAIN em 1986 (PCCH) especialmente, são muito diferentes, no aspecto metodológico e repercutiram bastante no meio científico. O objetivo do presente estudo foi de comparar métodos invasivos e não invasivos de avaliação aeróbia e anaeróbia em atletas ciclistas, identificando a relação existente entre o LM e a PC (determinada por dois métodos diferentes) e, a relação dos parâmetros anaeróbios, através da potência média e da potência pico com a CTA. Foram voluntários desse estudo 9 ciclistas treinados do sexo masculino (24,11 1,62 anos; 67,28 4,37 kg e 175,78 5,89 cm) com experiência na modalidade. Os atletas foram submetidos a três protocolos experimentais: 1) Teste de LM, com teste de Wingate para induzir a acidose e posterior exercício progressivo; 2) Teste de PCJQ com quatro sessões de cargas exaustivas randômicas; 3) Teste de PCCH a partir da variação da freqüência cardíaca e do lactato através de dois esforços de 180 segundos com intervalo de pausa de 90 segundos entre os esforços, com intensidade de 85, 95, 105 e 115% do LM. Foram encontradas diferenças significativas entre as intensidades de LM (219,73 4,43 W), PCJQ nos modelos Pot- 1/tlim... (Resumo completo, clicar acesso eletrônico abaixo). / Abstract: The anaerobic threshold (AT) has been used as a good predictor of the aerobic capacity for physical evaluation and training prescription. Among the protocols of AT determination, the lactate minimal test (LMT) seems to be one of the most accurate for obtain individually the AT values. Another model used in the aerobic evaluation is the non-invasive method for determination of the critical power (CP), and this model also makes possible the quantification of the intramuscular anaerobic stores, which represents the anaerobic work capacity (AWC). Several authors proposed determinations of CP on different protocols, as those propored by, Jenkins & Quigley (1991 - CPJQ) and Chassain (1986 - CPCH), that in spite of the very different methodological aspects they are a reference in the scientific way. The purpose of the present study was to compare invasive and non-invasive protocols to evaluate the aerobic and anaerobic fitness (by two different methods) in cyclists. Also, the relationship of the mean and peak power obtained during an Wingate test to AWC identified by different methods was investigate. Nine male trained cyclists volunteered to the study (24.11 l 1.62 years; 67.28 l 4.37 kg and 175.78 l 5.89 cm). The athletes were submitted to three experimental protocols: 1) LMT, with initial lactic acidosis induced thought a Wingate test and a subsequent incremental exercise test; 2) test of PCJQ with four sessions of random exhaustive loads; 3) test of PCCH based on the variation of the heart rate and lactate along two bouts of efforts of 180 seconds with pause interval of 90 seconds between them, with intensity of 85, 95, 105 and 115% of LMT. Our results showed significant differences among the intensities of LMT (219.73 l 4.43 W), PCJQ in the models P-1/tlim (301.47 l 10.46 W) and Wlim - tlim (295.22 l 10.14 W) and, in PCCH for the methods of lactate variation (201.53 l 4.44 W) and... (Complete abstract, click electronic address below). / Mestre

O objetivo desse estudo foi averiguar a possibilidade de se estimar os componentes anaeróbios do déficit máximo acumulado de oxigênio (MAOD) utilizando apenas o consumo de oxigênio (VO2). Adicionalmente, foi proposto um método alternativo que não necessita utilizar a extrapolação do VO2 do exercício submáximo para estimar a demanda de O2 para o exercício supramáximo. Para tanto, dez sujeitos fisicamente ativos foram submetidos aos seguintes testes: a) teste progressivo até a exaustão para determinar a carga correspondente ao consumo de oxigênio (WVO2max); b) seis testes de cargas constantes com intensidades abaixo da WVO2max (40-90% WVO2max); c) um teste a 110% da WVO2max; d) teste anaeróbio de Wingate. A fração do metabolismo anaeróbio alático foi estimada pela fase rápida do excesso do VO2 após o exercício (EPOCRÁPIDO), ao passo que a fração do metabolismo anaeróbio lático foi determinada tanto pela diferença entre MAOD e o EPOCRÁPIDO (MAODLA-1) como pelo acúmulo de lactato no sangue (MAODLA-2). O MAODMOD foi calculado pela somatória do EPOCRÁPIDO com MAODLA-2. Os principais resultados foram: a) as estimativas das contribuições do metabolismo anaeróbio lático não eram estatisticamente diferentes (MAODLA-1 = 2,40 ± 0,66 l; MAODLA-2 = 2,20 ± 0,4 l; p > 0,05); b) similarmente, os resultados do MAOD (3,03 ± 0,62 l) MAODMOD (2,84 ± 0,41) não eram significativamente diferentes (p > 0,05); c) foram detectadas correlações positivas entre MAODMOD, EPOCRÁPIDO, MAODLA-2 e os índices do teste de Wingate; d) os valores percentuais do EPOCRÁPIDO e do MAODLA-1 eram 22 ± 9% e 78 ± 9%, respectivamente. Esses resultados sugerem que os componentes anaeróbios do MAOD podem ser estimados satisfatoriamente utilizando somente o VO2 e que o MAOD pode ser determinado mediante MAODmod / The objective of this study was ascertain the possibility of estimate the anaerobic components of the maximal accumulated oxygen deficit (MAOD) used only oxygen uptake (VO2). In addition, was proposed an alternative method (MAODMOD) to determine the MAOD without using the extrapolation from submaximal VO2 to estimate the O2 demand of supramaximal exercise. Thus, ten subjects physically active performed the following tests: a) a maximal incremental exercise test for the measurement of the power output correspondent to maximal oxygen uptake (WVO2max); b) six submaximal tests with intensities bellow of the WVO2max (40-90% WVO2max); c) a supramaximal test at 110% WVO2max; d) Wingate anaerobic test. The fraction of the anaerobic alactic metabolism was calculated based on the fast component of excess post-exercise oxygen uptake (EPOCRÁPIDO), although the fraction of the anaerobic lactic metabolism was calculated based on both difference between MAOD and EPOCRÁPIDO (MAODLA-1) and changes in net blood lactate accumulation (MAODLA-2). MAODMOD was calculated by the sum of the EPOCRÁPIDO with the MAODLA-2. The main results were: a) non significant difference was found between the anaerobic lactic estimates (MAODLA-1 = 2,40 ± 0,66 l; MAODLA-2 = 2,20 ± 0,4 l; p > 0,05); b) similarly, there was no significant difference (p > 0,05) between MAOD (3,03 ± 0,62 l) and MAODMOD (2,84 ± 0,41); c) positive and significant correlation were detected between MAODMOD, EPOCRÁPIDO, MAODLA-2 and the indices of the Wingate anaerobic test (p < 0,05); d) the mean of the percentage values of the EPOCRÁPIDO and MAODLA-1 were 22 ± 9% e 78 ± 9%, respectively. These data suggest that the anaerobic components of the MAOD can be satisfactorily estimated using only the VO2 and that the MAOD can be determined using the MAODmod

O objetivo do presente trabalho foi realizar estudos acerca da degradação anaeróbia de proteínas no tocante à cinética de consumo de substrato e às rotas metabólicas presentes no processo. Inicialmente, realizou-se um estudo comparativo da degradação de efluentes sintéticos em reatores anaeróbios horizontais de leito fixo (RAHLFs) em escala de bancada. Nesse intuito, foi proposto um modelo cinética de primeira ordem de reações em série e paralelo, que permitiu a estimativa dos parâmetros cinéticos das várias etapas do processo estudado. As fontes de carbono nos efluentes sintéticos foram primeiramente compostas por compostos protéicos, tendo sido posteriormente adicionadas fontes de carboidratos e lipídeos à sua composição. Nesse intuito, utilizaram-se duas proteínas, caseína e soro albumina bovina (S.A.B.), e um hidrolisado protéico, peptona. Os resultados preditos pelo modelo cinético proposto apresentaram boa correlação com os dados experimentais e, com isso, foi possível o entendimento dos vários aspectos envolvidos na degradação de compostos protéicos em reatores que apresentam regime de escoamento predominantemente pistonado. A peptona foi mais rapidamente consumida, seguida pela caseína e finalmente pela S.A.B.. A presença de lipídeos nos efluentes sintéticos utilizados se constituiu no maior problema para o processo estudado. Análises de microscopia e biologia molecular mostraram que houve realocação da biomassa nos RAHLFs à medida que foram adicionadas fontes alternativas de carbono aos efluentes sintéticos à base de proteína. As morfologias encontradas foram comuns em todos os ensaios realizados, tendo predominado as semelhantes a Methanosaeta sp., bacilos fluorescentes e não fluorescentes, bacilos curvos e cocos não fluorescentes. Também foram visualizados três tipos de filamentos, espiroquetas e morfologias semelhantes a Methanosarcina sp. A segunda etapa do presente trabalho foi o estudo comparativo do processo de degradação de água residuária proveniente de abatedouro de aves em dois reatores um UASB e um RAHLF. O modelo cinético de primeira ordem se ajustou bem aos dados de decaimento de matéria orgânica em ambos os reatores. Dessa forma, foi possível concluir que no reator UASB, o mecanismo de adsorção do material colóide e solúvel nos grânulos parece ter regulado o processo, resultando em maior velocidade de remoção dessas frações de matéria orgânica. No RAHLF, o que parece ter regulado o processo foi a retenção de sólidos orgânicos, aumentando, dessa forma, a velocidade de remoção dessa fração de matéria orgânica. / This work reports on the anaerobic degradation of proteins from the standpoint of substrate consumption kinetics and metabolic routes. Initially, a comparative study of synthetic effluents in a bench scale horizontal-flow anaerobic immobilized biomass (HAIB) reactor was carried out. For this purpose, a kinetic model of irreversible first-order series-parallel reactions with two intermediate products was proposed. This allowed the estimative the kinetic parameters of several steps of the process. The carbon source in the synthetic effluents was composed primarily of casein, peptone and bovine serum albumin (B.S.A.). Then, sources of carbohydrate and lipid were added to the composition. The proposed model fitted the experimental protein degradation data well and allowed the best comprehension of the process. Thus it was possible to understand several aspects about the degradation of the protein substrates utilized in HAIB reactors. In relation to degradation rate, it was concluded that peptone was the most rapidly consumed, followed by casein, and finally BSA. Regarding substrate composition, the major problem for the studied process was the presence of lipids in the synthetics effluents. The results obtained from the microbiological analysis showed that relocation of biomass in the HAIBs reactors happened in correlation with the addition of alternative carbon sources to synthetic protein effluents. In all assays, common morphologies were found. These werepredominantly Methanosaeta sp. - like archaeas, fluorescent bacillus (three types), non-fluorescent bacillus (6 types), curved bacillus and non-fluorescent coccus. Three types of filaments, spirochetes and Methanosarcina sp. - like archaeas were also found. In a second phase, a comparative analysis of the degradation process of wastewater from a poultry slaughterhouse was carried out in two different reactors, one up flow anaerobic sludge blacked (UASB), and one HAIB. The first order kinetic model proposed fitted the experimental organic matter degradation data well in UASB and HAIB reactors treating residual water from avian abattoirs. In the UASB reactor, the adsorption mechanism of the granules for colloid and soluble material seems to regulate the process. This results in a higher rate of removal for these fractions of organic material. However, in the HAIB reactor, what seems to be controlling the process was the retention of organic solids, which increased the removal rate for this fraction of organic material.

Submitted in complete fulfillment for the degree of Doctor of Philosophy (Biotechnology), Durban University of Technology, Durban, South Africa, 2015. / Anaerobic digestion, a proven and highly efficient biological process for treating industrial wastewater and biogas generation is an underutilized technology in South Africa. Some of the industries that have on-site anaerobic reactors tend to face problems in operating these reactors due to poor understanding of the process and implementation of the technology. This has resulted in high pollutant loads in their final effluents and low energy recovery. In this study, an on-site full–scale upflow anaerobic sludge blanket (UASB) reactor treating brewery wastewater was extensively monitored in order to evaluate the efficiency in terms of effluent quality, biogas production and microbial structure. Furthermore, developed and adopted kinetic models were used to optimize the performance of the full–scale UASB reactor using a combined Pareto differential evolution (CPMDE) algorithm. A preliminary analysis of the raw wastewater has shown that the wastewater produced from the brewery industry was high in organic matter with a total chemical oxygen demand (COD) between 1096.41 to 8926.08 mg/L. The average removal efficiency of COD from the UASB reactor after treatment was 79% with a methane (CH4) production of 60-69% at temperature ranges of 28-32˚C and hydraulic retention time (HRT) of 12 h within the optimal pH range for anaerobic bacteria (6.6 and 7.3) under various organic loading rates. However, the results also showed an increase in total suspended solids (TSS), nitrogen (N2), ammonia (NH3) and orthophosphate concentrations when comparing the influent to the effluent, which indicated the necessity for further optimization of the reactor condition in order to reduce these effluent parameters to acceptable standards and to increase CH4 production. In order to optimize the process, a thorough understanding of microbial interaction was essential. A combination of different molecular techniques viz., fluorescence in–situ hybridization (FISH), polymerase chain reaction (PCR) and quantitative real-time PCR (QPCR) were employed to understand the microbial community structure of the granular sludge samples using species specific primers and probes. The results revealed that the dominance of diverse groups of eubacteria belonging to phyla Proteobacteria, Firmicutes and Chloroflexi and an uncultured candidate division WS6 with four different orders of methanogenic Archaea viz., Methanomicrobiales, Methanococcales, Methanobacteriales and Methanosarcinales belonging to hydrogenotrophic and aceticlastic methanogens were within the reactor samples. Quantification of the 16S rDNA copies of eubacteria and methanogenic Archaea using species-specific primers further confirmed the spatial distribution of these microorganisms within the different compartments of the reactor where, the upper compartments were dominated by eubacteria and the lower compartments by methanogenic Archaea. The concentration of Archaea per nanogram of DNA was much higher (96.28%) than eubacteria (3.78%) in lower compartments, while, the eubacteria concentration increased to 98.34% in upper compartments with a decrease in Archaea quantity (1.66%). A modified kinetic methane generation model (MMGM) was developed on the basis of mass balance principles with respect to substrate (COD) degradation and the endogenous decay rate to predict CH4 production efficiency of the reactor. Furthermore, a Stover–Kincannon kinetic model was adopted with the aim of predicting the final effluent quality in terms of COD concentration and model coefficients were determined using the data collected from the full–scale reactor. Thereafter, a model-based multi-objective optimization was carried out using the CPMDE algorithm with three–objective functions namely; maximization of volumetric CH4 production rate; minimization of effluent substrate concentration and minimization of biomass washout, in order to increase the overall efficiency of the UASB reactor. Important decision variables and constraints related to the process were set for the optimization. A set of non-dominated solutions with high CH4 production rates of between 2.78 and 5.06 L CH4/g COD/day at low biomass washout concentrations were obtained at almost constant solution for the effluent COD concentration. A high COD removal efficiency (85-87%) at ~30-31˚C and 8-9 h HRT was obtained for the multi-objective optimization problem formulated. This study could significantly contribute towards optimization of a full–scale UASB reactor treating brewery wastewater for better effluent quality and biogas production. Knowledge on the activity and performance of microbial community present in the granular sludge taken from the full–scale UASB reactor would contribute significantly to future optimization strategies of the reactor. In addition, optimization using an evolutionary algorithm under different operational conditions would help to save both time and resources wasted in operating anaerobic bioreactors.

Le processus de dégradation anaérobie de la matière organique est un phénomène naturel largement répandu sur terre (ex. marais, lacs, rizières, systèmes digestifs d'animaux et humains). Une très grande diversité microbienne est entretenue durant ce processus, traduisant une diversité de voies métaboliques impliquées. Lorsqu'elle est complète, la digestion anaérobie aboutie à la formation de biogaz (mélange de méthane et de dioxyde de carbone). En termes de biotechnologie, le traitement par voie anaérobie de pollutions organiques permet de réduire le volume de déchets en générant du méthane valorisable sous plusieurs formes (électricité, chaleur, gaz naturel, biocarburant). Cependant, les digesteurs industriels sont optimisés pour un fonctionnement à 35°C ou à 55°C, ce qui nécessite un apport exogène d'énergie de maintenance. Ainsi, les travaux de thèse se sont intéressés à l'étude de la capacité d'adaptation de divers écosystèmes anaérobies industriels couvrant une variété de procédés et de conditions opératoires à convertir l'éthanol en biogaz à différentes températures. La première phase de l'étude avait pour but le conditionnement, en réacteurs de laboratoire d'écosystèmes à leur température d'origine avec un substrat facilement biodégradable (éthanol). Ensuite, les performances des communautés microbiennes (le potentiel méthanogène maximal et la cinétique de dégradation) ont été estimées sur un gradient de température de 5°C à 55°C en fioles. La phase de conditionnement des écosystèmes en réacteur batch a montré que la production de biogaz avoisinait la production théorique et que cette production s'accompagnait d'une diminution de la durée de réaction avec ajout successif du substrat. De plus, les cinétiques de production de biogaz obtenues les variaient fortement d'un écosystème à l'autre. Des profils d'empreintes moléculaires (CE-SSCP) des communautés bactériennes et archées ont été réalisés au début et à la fin du conditionnement. Ces profils de communauté ont été comparés entre eux par analyse en composante principale (ACP). Les populations bactériennes qui assuraient une performance efficiente étaient différentes de celles qui garantissaient une bonne capacité d'adaptation. Par ailleurs, le potentiel d'adaptation dépendait de la présence de populations d'Archaea méthanogènes bien spécifiques. En plaçant ensuite les écosystèmes conditionnés dans des conditions de température éloignées de la température d'origine, seuls les écosystèmes mésophiles se sont acclimatés aux températures psychrophiles. Comme attendu, l'activé spécifique maximale des méthanogènes était toujours obtenue à la température d'origine de l'écosystème. L'analyse des communautés bactériennes et archées à la fin de la période d'acclimatation a révélé que l'acclimatation des écosystèmes thermophiles et mésophiles à des températures plus faibles ne modifiait que légèrement la structure des communautés microbiennes. En revanche, des changements plus importants étaient obtenus lorsque la température d'incubation était augmentée par rapport à la température d'origine de l'écosystème. En résumé, l'étude de l'effet de la température d'incubation (de 5°C à 55°C) sur l'activité fermentaire et sur la structure des populations microbiennes est un bon modèle d'étude au laboratoire pour appréhender l'impact d'un facteur abiotique sur la dynamique structurelle et fonctionnelle d'une communauté microbienne complexe. / The process of anaerobic degradation of organic matter is a natural phenomenon widespread on Earth (eg, marshes, lakes, rice fields, digestive systems of animals and humans). A high microbial diversity is maintained during this process, reflecting a diversity of metabolic pathways involved. When complete, the anaerobic digestion accomplished in the formation of biogas (methane mixture and carbon dioxide). In terms of biotechnology, anaerobic treatment of organic pollution reduces the volume of waste and generates methane recoverable in several forms (electricity, heat, natural gas, biofuels). However, industrial digesters are optimized for operation at 35 ° C or 55 ° C, which requires exogenous energy maintenance. Thus, the thesis is interested in the study of the adaptability of various anaerobic ecosystems covering a variety of industrial processes and operating conditions to convert ethanol into biogas at different temperatures. The first phase of the study was to the conditioning, in laboratory reactors ecosystems to their original temperature with a readily biodegradable substrate (ethanol). Then, the performances of microbial communities (the maximum methanogenic potential and degradation kinetics) were estimated on a temperature gradient of 5 ° C to 55 ° C in glass bottles. The conditioning phase of the ecosystems in batch reactor showed that the biogas averaged theoretical production and this production was followed by a decrease in reaction time with successive addition of the substrate. In addition, the kinetics of the biogas obtained varied greatly from one ecosystem to another. Molecular fingerprinting profiles (CE-SSCP) of bacterial and archaeal communities were performed at the beginning and at the end of conditioning. These community profiles were compared with each other by principal component analysis (PCA). Bacterial populations that ensured efficient performance were different from those that ensured a good adaptability. In addition, the potential for adaptation depended on the presence of very specific methanogenic Archaea populations. When placing ecosystems conditioned in temperature away from the original temperature, only mesophilic ecosystems adapted to psychrophilic temperatures. As expected, specific methanogenic activity was always obtained at the original temperature of the ecosystem. Analysis of bacterial and archaeal communities at the end of the acclimation period revealed that acclimation thermophilic and mesophilic ecosystems to lower temperatures only modified slightly the structure of microbial communities. On the other hand, more significant changes were obtained when the incubation temperature was increased in comparison to the original temperature of the ecosystem. In summary, the study of the effect of incubation temperature (5 ° C to 55 ° C) on the fermentation activity and microbial population structure is a good model for laboratory study to understand the impact of abiotic factor on the structural and functional dynamics of a complex microbial community.

Para minimizar os impactos ambientais causados pelo lançamento de formaldeído na natureza, é importante o desenvolvimento de tecnologias adequadas para o seu tratamento. Sistemas aeróbios têm sido utilizados, porém, as opções por tratamento anaeróbio estão tendo cada vez mais aceitação, principalmente pelo baixo consumo de energia e baixa produção de lodo. Por outro lado, substâncias tóxicas podem provocar distúrbios em reatores anaeróbios. Algumas pesquisas já foram desenvolvidas quanto ao tratamento biológico anaeróbio de formaldeído, no entanto, ainda não se chegou a um consenso sobre o seu comportamento ou qual o sistema mais eficaz. Nesse trabalho, avaliou-se a degradação e a toxicidade dessa substância em reator anaeróbio horizontal de leito fixo, a fim de trazer subsídios para essa questão. As concentrações de estudo variaram de, aproximadamente, 30 a 1500 mg/L de formaldeído, atingindo-se as eficiências de remoção de até 99,7 % de formaldeído e 92 % de DQO. A presença de ácidos graxos com até cinco carbonos durante a degradação do formaldeído pode indicar uma rota de degradação diferente da sugerida na literatura, onde os produtos intermediários seriam o metanol e o ácido fórmico. O modelo cinético de Monod foi o que melhor representou os dados experimentais, com \'R IND.max\' POT.app\' = 2,79 x \'10 POT.-3\' HCHO/mg SSV.h e \'K IND.S\' POT. app\' = 242,8 mg HCHO/L. / In order to minimize the environmental impacts caused by formaldehyde discharge in the nature, the development of appropriate technologies for its treatment is important. Aerobic systems have been used, however, anaerobic treatment options have been widely studied, mainly due to low energy consumption and sludge production. On the other hand, toxic substances can lead to disturbances in anaerobic reactors. Some formaldehyde anaerobic biological treatment researches have already been developed, although, there is no consensus about its behavior or which is the most efficient system. Degradation and toxicity of this substance were evaluated in horizontal-flow anaerobic immobilized sludge reactor, with the purpose of producing subsidies for this issue. Formaldehyde concentrations ranging from, approximately, 30 to 1500 mg/L were applied in the reactor and formaldehyde and COD removal efficiencies of 99,7% e 92% were achieved, respectively. Volatile fatty acids with up to 5 carbons, found during the formaldehyde degradation, can indicate that degradation followed different routes than those suggested in the literature, that reports the formation of intermediates such as methanol and formic acid. Monod kinetic model fitted well the experimental data, with apparent kinetic parameters estimated as \'R IND.max\' POT. app\' = 2,79 x \'10 POT.-3\' mg HCHO/mg SSV.h and \'K IND.S\' POT. app\' = 242,8 mg HCHO/L.