Submitted by Sara Ribeiro (sara.ribeiro@ucb.br) on 2018-11-07T17:34:33Z No. of bitstreams: 1 HenriqueCesarTeixeiraVerasDissertacao2013.pdf: 26147875 bytes, checksum: 3014f6ec0d791f46e4e1ee8c49894c2f (MD5) / Approved for entry into archive by Sara Ribeiro (sara.ribeiro@ucb.br) on 2018-11-07T17:35:09Z (GMT) No. of bitstreams: 1 HenriqueCesarTeixeiraVerasDissertacao2013.pdf: 26147875 bytes, checksum: 3014f6ec0d791f46e4e1ee8c49894c2f (MD5) / Made available in DSpace on 2018-11-07T17:35:09Z (GMT). No. of bitstreams: 1 HenriqueCesarTeixeiraVerasDissertacao2013.pdf: 26147875 bytes, checksum: 3014f6ec0d791f46e4e1ee8c49894c2f (MD5) Previous issue date: 2013-09-06 / Humans live in constant association with microorganims. The amount of microorganims present in the human body exceeds our own cell number. That community of microorganisms has deep influence in health and disease. The use of high-throughput DNA sequencing technologies and culture independent approaches have been enlarging the understanding concerning the communities of microorganisms and the association of these with the host. The human gastrointestinal tract contains one of the most complex bacterial communities. It was proposed recently that the microbiome can be classified in three enterotypes. In our study, we used data metagenomics quantitative search to identify phylogenetic patterns in the intestinal microbiome to develop prediction models for the enterotypes. To reach this aim, statistical tests were applied to the data regarding abundance of bacteria in level taxonomic corresponding to genus. We identified genus significantly with the abundance different and important correlations. Besides the ratio among genus to be used as parameter bioindicator of the respectives enterotypes. Through the logistic regression test we identified that the prediction model for ET1 was influenced significantly by the ratio of Bacteroides / (Prevotella + Ruminococcus). In the model for prediction of ET2, it was the ratio of Prevotella / Bacteroides with such as characteristic significance. And for the model of ET3, we identified the ratio of (Akkermansia + Alistipes) / (Bacteroides + Prevotella) as significant parameter. These models were assessed against two groups of independent data and associated with the value of cut-off 5%; 20% and 95% respectively. Besides the value of cutoff for each models, the crossed validation allowed the association of the model with the measures of PPV for ET1, specificity for ET2 and PNV for ET3. We propose the experimental validation of these models for the qPCR technique. And with that methodology established, it would be possible to do the diagnosis of the enterotype individually. / Humanos vivem em constante associação com microrganismos. A quantidade de microrganismos presentes no corpo humano ultrapassa o nosso próprio número de células. Essa comunidade de microrganismos tem profunda influência na saúde e doenças. As tecnologias de sequenciamento de DNA de alta capacidade e abordagens moleculares independente de cultura têm ampliado a compreensão acerca das comunidades de microrganismos e a associação destes com o hospedeiro. O trato gastrointestinal humano abriga uma das mais complexas comunidades bacterianas. Foi proposto recentemente que o microbioma pode ser categorizado em três enterotipos. No nosso estudo, utilizamos dados metagenômicos quantitativos buncando identificar padrões filogenéticos no microbioma intestinal para desenvolver modelos de predição para os enterotipos. Para alcançar este objetivo, testes estatísticos foram aplicados aos dados referente a abundância de bactérias em nível taxonômico correspondente a gênero. Identificamos gêneros com a abundância significativamente diferente e correlações importantes. Além da razão entre gêneros para ser utilizada como parâmetro bioindicativo dos respectivos enterotipos. Através do teste de regressão logística identificamos que o modelo de predição para o ET1 foi influenciado significativamente pela razão de Bacteroides / (Prevotella + Ruminococcus). No modelo para predição do ET2, foi a razão de Prevotella / Bacteroides que apresentou significância. E para o modelo do ET3, identificamos a razão de (Akkermansia + Alistipes) / (Bacteroides + Prevotella) como parâmetro significativo. Estes modelos foram avaliados contra dois conjuntos de dados independentes e associados com o valor de cut-off 5%; 20% e; 95% respectivamente. Além do valor de cut-off para cada modelos, a validação cruzada permitiu a associação do modelo com as medidas de PPV para o ET1, especificidade para o ET2 e PNV para o ET3. Propomos a validação experimental destes modelos pela técnica de qPCR. E com essa metodologia estabelecida, seria possível fazer o diagnóstico do enterotipo individualmente.

Introdução: o estresse psicológico aumenta a permeabilidade intestinal em roedores e humanos, potencialmente levando a inflamação de baixo grau e aos sintomas em distúrbios gastrintestinais funcionais. No entanto, o efeito do estresse agudo combinado ao estresse da vida crônica, que mimetiza potencialmente melhor a situação humana, é desconhecido. Além disso, há poucos dados disponíveis sobre os efeitos do estresse em intestino delgado versus cólon. Métodos: ratos Wistar foram alocados em quatro protocolos de estresse: 1/ controles; 2/ estresse agudo (isolamento e movimentos limitados); 3/ Crowding stress:crônico e 4/ estresse agudo + estresse crônico. Amostras de jejuno e cólon foram colhidas para estudar a permeabilidade em câmaras deUssing, a expressão gênica de moléculas de junção firmes e a densidade de mastócitos. Níveis de corticosterona no plasma foram medidos. Principais resultados:corticosterona plasmática foi avaliada nas três condições de estresse, teve níveis mais altos na condição de estresse combinado. Permeabilidade do jejuno foi aumentada em todas as condições de estresse e correlacionada com os níveis de corticosterona. O aumento da expressão das claudinas 1, 5 e 8, daocludina e da ZO-1 foi detectado no estado de estresse agudo no jejuno. Em contraste, a permeabilidade do cólon foi aumentada no protocolo de estresse combinado, e a expressão de moléculas das junção firmes permaneceu inalterada. O aumento da densidade de mastócitos foi observado no cólon nos ratos submetidos aos estresses crônico e combinado. Conclusão e inferências:os estresses agudo, crônico e combinado influenciam diferentemente a permeabilidade intestinal, a expressão de moléculas de junção firmes e a atividade dos mastócitos, no jejuno e no cólon. Estes resultados fornecem uma visão mais aprofundada dos mecanismos de hiperpermeabilidade intestinal relacionadas ao estresse. / Background: Psychological stress increases intestinal permeability in rodents and humans, potentially leading to low-grade inflammation and symptoms in functional gastrointestinal disorders through disturbances in brain-gut axis. However, the effect of acute stress on the background of Crhonic life stress, potentially better approaching the human situation, is unknown. Moreover, only limited information is available on the effects in small intestine versus colon in animal model. Methods: Wistar rats were allocated to 4 stress protocols: 1/ sham; 2/ acute stress (isolation and limited movement); 3/ Crhonic crowding stress and 4/ acute + Crhonic stress (n = 8 per group). Jejunum and colon were harvested to study permeability in Ussing chambers, gene expression of tight junction molecules and mast cell density. Plasma corticosterone levels were measured. Key Results: Plasma corticosterone was elevated in all three stress conditions, with the highest levels in the combined stress condition. Permeability of the jejunum was increased in all stress conditions and correlated with corticosterone levels. Increased expression of claudin 1, 5 and 8, occludin and ZO-1 was detected in the acute stress condition in the jejunum. In contrast, colonic permeability was increased in the acute on Crhonic stress protocol only and the expression of tight junction molecules was unaltered. Increased mast cell density was observed in the Crhonic and acute on Crhonic stress condition in the colon only. Conclusion and Inferences: Acute, Crhonic and combined stress differentially affect intestinal permeability, expression of tight junction molecules and mast cells in the jejunum and the colon. These findings provide further insight in the mechanisms of stress-related intestinal hyperpermeability and barrier.

O objetivo deste estudo foi avaliar o efeito da mistura comercial do líquido da casca de castanha de caju e do óleo de mamona (Essential, Oligo Basics Agroind. Ltda., Cascavel, Brasil) no desempenho, na microbiota e no sistema imune de frangos de corte desafiados ou não por coccidiose. Ao total 864 pintos machos (Cobb) de um dia de idade foram distribuídos aleatoriamente em 6 tratamentos (8 boxes/tratamento e 18 pintos/box) em um desenho fatorial 3 x 2 com 3 aditivos: controle (sem aditivo), 100 ppm de monensina ou 0,15% de Essential e 2 níveis de desafio aos 14 dias de idade: não desafiados ou inoculados por gavagem com 1mL de solução contendo oocistos esporulados de E. tenella, E. acervulina e E. máxima. Os resultados foram divididos em dois artigos. Artigo 1: Na primeira semana após desafio, as aves desafiadas suplementadas com monensina apresentaram maior ganho de peso (GP), consumo de ração (CR) e melhor conversão alimentar (CA) (P <0,05), porém na segunda semana o Essential apresentou maior GP e melhor CA (P <0,05), aos 42 dias de idade, ambos os grupos não se diferiram em GP, CR, PV e foram maiores do que o controle (P <0,05). A utilização de monensina em aves desafiadas reduziu o número cópias do domínio bactéria e de E.coli (P<0,05), por sua vez, a suplementação com Essential reduziu Clostridium Cluster XIV, Clostridium perfringens e Staphylococcus aureus em relação aos demais tratamentos (P<0,05). As aves não desafiadas que receberam Essential ou monensina apresentaram menor população de C.perfringens e S. aureus (P<0,05). Artigo 2: O grupo que recebeu Essential aumentou a expressão gênica de IFN-y, IL-6 e TNF-α (P<0,05) e o grupo controle aumentou a expressão gênica de COX-2 e IL-1 em relação aos demais tratamentos (P<0,05). As aves não desafiadas que receberam monensina apresentaram maior expressão gênica de IFN-y, COX-2 e IL-1 comparadas aos demais tratamentos (P<0,05), ao contrário do grupo com Essential que reduziu a expressão gênica com exceção do TNF-α. Aos 7 e 14 dias após o desafio houve maior excreção de oocistos para o grupo controle, Essential e monensina não diferiram-se (P>0,05). Assim, o Essential melhorou o desempenho de frangos de corte infectados por coccidiose após a segunda semana do desafio e atuou como um modulador da microbiota intestinal e do sistema imune, direcionando a resposta inflamatória contra o parasita. / The objective of this study was to evaluate the effects of a commercial mixture of cashew shell liquid and castor oil (Essential, Oligo Basics Agroind. Ltda., Cascavel, Brazil) on growth performance, immunity and microbiota in broilers challenged with coccidiosis. A total of 864 one-day-old male chicks (Cobb) were randomly assigned to 6 treatments (8 pens/treatment and 18 birds/pen) in a 3 x 2 factorial design with 3 additives: control (no additive), 100 ppm of monensin, and 0.15% of Essential; and 2 challenge levels at 14 days of age: no challenge and inoculation by gavage of 1 ml of a solution containing sporulated oocysts of E. tenella, E. acervulina, and E. maxima. The results were divided into two articles. Article 1: In the first week after challenge, challenged birds supplemented with monensin showed higher LW, WG, FI and better FCR (P <0.05), but in the second week Essential presented higher WG and better FCR (P <0.05), at 42 days of age, both groups did not differ in WG, FI, and LW and were higher than the control (P <0.05). The use of monensin in challenged birds reduced the number of copies of the bacteria domain and of E.coli (P<0,05). In turn, Essential supplementation reduced Clostridium Cluster XIV, Clostridium perfringens and Staphylococcus aureus in relation to the other treatments (P<0,05). The unchallenged birds that received Essential or monensin presented a lower population of C.perfringens and S. aureus (P<0,05). In addition, Essential presented higher number of copies of Lactobacillus spp., followed by monensin and control (P <0.05). Article 2: The group that received Essential increased the gene expression of IFN-y, IL-6 e TNF-α (P<0,05) and the control group increased the gene expression of COX-2 and IL-1 in relation to the other treatments (P <0.05). The unchallenged birds that received monensin presented upregulated expression of IFN-y, COX-2 and IL-1 compared to the other treatments (P <0.05), unlike the Essential group, which reduced gene expression with the exception of TNF-α. At 7 and 14 days after the challenge there was a higher excretion of oocysts for the control group, Essential and monensin did not differ (P>0,05). Thus, Essential improved the performance of coccidiosis-infected broiler chickens after the second week of challenge, as well as acts as a modulator of intestinal flora and immune system, directing the inflammatory response against the parasite.

O presente estudo busca fornecer subsídios para a evolução da qualidade de seleção da fiscalização da Compensação Financeira da Exploração Mineral (CFEM) visando a propiciar um aumento na recuperação de créditos em cada fiscalização. Pretende-se mensurar os parâmetros que possam ser utilizados na definição da priorização de empresas a serem auditadas in loco, com a construção, através de pesquisa exploratória por meio de análise documental e entrevistas com especialistas do setor da matriz de priorização. Com esta pesquisa, espera-se contribuir na otimização da arrecadação das receitas públicas, assim como melhorar os resultados através do aumento da eficácia das fiscalizações, ou seja, aumentando o grau de acerto das fiscalizações e reduzindo o número de fiscalizações sem resultado. A Auditoria Fiscal realizada pelo DNPM em empresas de mineração pode ser dividida nas seguintes etapas: Seleção da empresa; Observação dos Processos Operacionais da Empresa; Inspeção da Documentação Contábil, Fiscal e Gerencial; Relatório de Fiscalização; Lavratura da Notificação Fiscal de Lançamento de Débitos para Pagamento (NFLPD). Nesta pesquisa, foram utilizados métodos primários (entrevistas com especialistas do setor) e secundários de pesquisa como a revisão da literatura. O método Delphi foi utilizado para coletar a opinião dos especialistas do setor. Quanto à técnica, utilizou-se a matriz GUT (Gravidade, Urgência, Tendência) que tem como objetivo estabelecer as prioridades das ações entre diversas alternativas, em que são atribuídos graus de importância nos valores de 1 a 5 para cada um dos problemas que compõe essa matriz. Na fase de entrevistas, identificou-se o nível de consenso e a prioridade dos critérios para seleção de empresas para fins de auditoria da CFEM. Concluiu-se que a adoção de critérios para direcionamento das auditorias externas da CFEM é mais eficiente do que a seleção aleatória realizada atualmente na SUP/DNPM/RS. Propõe-se que essa conclusão seja considerada em futuros planejamentos na SUP/DNPM/RS, para produzir melhorias em termos de arrecadação da CFEM e para tornar o processo de seleção eficiente. / The present study seeks to provide subsidies for the evolution of the selection quality of the Financial Compensation for Mineral Exploration (FCME) supervision in order to provide an increase in the recovery of credits in each inspection. It is intended to measure the parameters that can be used to define the prioritization of companies to be audited in loco, with the construction, through exploratory research through documentary analysis and interviews with specialists of the sector of the prioritization matrix. With this research, we hope to contribute to the optimization of the collection of public revenues, as well as to improve the results by increasing the efficiency of the inspections, that is, increasing the degree of correctness of the inspections and reducing the number of inspections without result. The Fiscal Audit carried out by DNPM in mining companies can be divided into the following stages: Company selection; Observation of the Company's Operating Processes; Inspection of Accounting, Tax and Management Documentation; Inspection Report; Drawing up of the Tax Notification of Debits for Payment (TNDP). In this research, primary (interviews with industry experts) and secondary research methods were used as the literature review. The Delphi method was used to gather the opinion of industry experts. As for the technique, the GUT (Severity, Urgency, Tendency) matrix was used to establish the priorities of the actions among several alternatives, in which degrees of importance are assigned in the values from 1 to 5 for each of the problems that compose this matrix. In the interview phase, the level of consensus and the priority of the criteria for company selection for FCME audit purposes were identified. It was concluded that the adoption of criteria to guide FCME's external audits is more efficient than the current random selection performed at SUP/DNPM/RS. It is proposed that this conclusion be considered in future SUP/DNPM/RS planning to produce improvements in terms of FCME collection and to make the selection process efficient.

Diante da importância do zinco no desempenho produtivo e reprodutivo de matrizes suínas, e no desempenho de leitões, foram conduzidos três experimentos com o objetivo de avaliar diferentes fontes de zinco na alimentação de suínos. O experimento I avaliou diferentes fontes de zinco, orgânico (zinco+metionina ((Zn-Met) e zinco+glicina(Zn-Gli)) e inorgânico (óxido de zinco(ZnO)) na dieta de matrizes em gestação e lactação e os efeitos do uso destas fontes na sua progênie. Foram utilizadas 18 fêmeas de uma linhagem comercial distribuídas em delineamento experimental de blocos ao acaso, com três tratamentos e seis repetições de um animal. As características avaliadas nas matrizes foram: peso, espessura de toucinho, prolificidade, número de leitões nascidos vivos e níveis séricos de zinco no colostro e no leite. Para os leitões após parto até a desmama foram analisados: níveis de zinco no soro, ganho de peso, histomorfologia entérica e densidade óssea. No experimento II foram utilizados 180 leitões, desmamados com 21 dias de idade, distribuídos em delineamento de blocos ao acaso em esquema fatorial 3 x 3 (3 dietas maternas - 100 ppm de ZnO, Zn-Met, Zn-Gli x 3 dietas da progênie - 100 ppm de ZnO, Zn-Met, Zn-Gli) totalizando 9 tratamentos e 5 repetições de 4 animais cada, em que foram avaliados o desempenho, incidência de diarréia, níveis de zinco no soro, densidade óssea e histomorfologia entérica No experimento III foram utilizados 810 leitões, desmamados com 21 dias de idade, distribuídos em delineamento de blocos ao acaso com 6 tratamentos de 3 repetições de 45 animais: Fases I e II - 1. dieta + 2300 ppm de ZnO; 2. dieta + 2200 ppm de ZnO + 100 ppm de Zn-Met; 3. dieta + 2200 ppm de ZnO + 100 ppm de Zn-Gli; 4. dieta + 2400 ppm de ZnO; 5. dieta + 2200 ppm de ZnO + 200 ppm de Zn-Met; 6. dieta + 2200 ppm de ZnO+200 ppm de Zn-Gli; Fase III - 1. dieta + 1600 ppm de ZnO; 2. dieta + 1500 ppm de ZnO + 100 ppm de Zn-Met; 3. dieta + 1500 ppm de ZnO + 100 ppm de Zn-Gli; 4. dieta + 1700 ppm de ZnO; 5. dieta + 1500 ppm de ZnO + 200 ppm de Zn-Met; 6. dieta + 2200 ppm de ZnO+200 ppm de Zn-Gli, em que avaliou-se o desempenho dos animais. Os dados foram analisados utilizando o programa SAS (1998). As médias dos tratamentos foram comparadas pelo teste de Tukey ao nível de 5% de probabilidade, segundo Steel e Torrie (1980). / Given the importance of zinc in the productive and reproductive performance of sows and piglets performance, three experiments to evaluate different sources of zinc in swine were conducted. The first experiment evaluated different sources of zinc, organic (zinc methionine (Zn-Met) and zinc glycine (Zn-Gly)) and inorganic (zinc oxide (ZnO)) in the diet of pregnant gilts and sows lactating and the effects of using these sources in their progeny. Were used 18 female of a commercial strain distributed in experimental design of randomized blocks with three treatments and six repetitions with one animal each. The characteristics evaluated in the sows were: weight, backfat thickness, prolificacy, number of piglets born alive and zinc levels in serum, colostrum and milk; In the piglets after birth until weaning were analyzed: serum levels of zinc, weight gain, bone density and histomorphology enteric. The second experiment 180 piglets were used, weaned at 21 days of age in a randomized block design in a factorial 3 x 3 (3 maternal diets - 100 ppm from ZnO , Zn-Met , Zn-Gly x 3 diets progeny - 100 ppm ZnO , Zn-Met - , Zn-Gly) totaling 9 treatments and 4 replicates of 5 animals each, which evaluated the performance, diarrhea incidence, serum zinc levels, bone density and histomorphology enteric. The third experiment 810 weaned pigs weaned at 21 days of age in a randomized block design with 6 treatments of 3 replicates of 45 animals each: Phases I and II - 1. diet + 2300 ppm of ZnO; 2. diet + 2200 ppm of ZnO + 100 ppm of Zn-Met; 3. diet + 2200 ppm ZnO + 100 ppm of Zn-Gly; 4. diet + 2400 ppm of ZnO; 5. diet + 2200 ppm of ZnO + 200 ppm of Zn- Met; 6. diet + 2200 ppm of ZnO + 200 ppm of Zn-Gly; Phase III - 1. diet + 1600 ppm of ZnO; 2. diet + 1500 ppm of ZnO + 100 ppm of Zn-Met; 3. diet + 1500 ppm of ZnO + 100 ppm of Zn-Gly; 4. diet + 1700 ppm of ZnO; 5. diet + 1500 ppm of ZnO + 200 ppm of Zn- Met; 6. diet + 2200 ppm of ZnO + 200 ppm of Zn-Gly in evaluating the performance of animals. All datas were analyzed using SAS (1998 ) program. The treatment means were compared by Tukey test at 5 % probability , according to Steel and Torrie (1980).

abstract: Machine learning (ML) has played an important role in several modern technological innovations and has become an important tool for researchers in various fields of interest. Besides engineering, ML techniques have started to spread across various departments of study, like health-care, medicine, diagnostics, social science, finance, economics etc. These techniques require data to train the algorithms and model a complex system and make predictions based on that model. Due to development of sophisticated sensors it has become easier to collect large volumes of data which is used to make necessary hypotheses using ML. The promising results obtained using ML have opened up new opportunities of research across various departments and this dissertation is a manifestation of it. Here, some unique studies have been presented, from which valuable inference have been drawn for a real-world complex system. Each study has its own unique sets of motivation and relevance to the real world. An ensemble of signal processing (SP) and ML techniques have been explored in each study. This dissertation provides the detailed systematic approach and discusses the results achieved in each study. Valuable inferences drawn from each study play a vital role in areas of science and technology, and it is worth further investigation. This dissertation also provides a set of useful SP and ML tools for researchers in various fields of interest. / Dissertation/Thesis / Doctoral Dissertation Electrical Engineering 2018

Diante da importância do zinco no desempenho produtivo e reprodutivo de matrizes suínas, e no desempenho de leitões, foram conduzidos três experimentos com o objetivo de avaliar diferentes fontes de zinco na alimentação de suínos. O experimento I avaliou diferentes fontes de zinco, orgânico (zinco+metionina ((Zn-Met) e zinco+glicina(Zn-Gli)) e inorgânico (óxido de zinco(ZnO)) na dieta de matrizes em gestação e lactação e os efeitos do uso destas fontes na sua progênie. Foram utilizadas 18 fêmeas de uma linhagem comercial distribuídas em delineamento experimental de blocos ao acaso, com três tratamentos e seis repetições de um animal. As características avaliadas nas matrizes foram: peso, espessura de toucinho, prolificidade, número de leitões nascidos vivos e níveis séricos de zinco no colostro e no leite. Para os leitões após parto até a desmama foram analisados: níveis de zinco no soro, ganho de peso, histomorfologia entérica e densidade óssea. No experimento II foram utilizados 180 leitões, desmamados com 21 dias de idade, distribuídos em delineamento de blocos ao acaso em esquema fatorial 3 x 3 (3 dietas maternas - 100 ppm de ZnO, Zn-Met, Zn-Gli x 3 dietas da progênie - 100 ppm de ZnO, Zn-Met, Zn-Gli) totalizando 9 tratamentos e 5 repetições de 4 animais cada, em que foram avaliados o desempenho, incidência de diarréia, níveis de zinco no soro, densidade óssea e histomorfologia entérica No experimento III foram utilizados 810 leitões, desmamados com 21 dias de idade, distribuídos em delineamento de blocos ao acaso com 6 tratamentos de 3 repetições de 45 animais: Fases I e II - 1. dieta + 2300 ppm de ZnO; 2. dieta + 2200 ppm de ZnO + 100 ppm de Zn-Met; 3. dieta + 2200 ppm de ZnO + 100 ppm de Zn-Gli; 4. dieta + 2400 ppm de ZnO; 5. dieta + 2200 ppm de ZnO + 200 ppm de Zn-Met; 6. dieta + 2200 ppm de ZnO+200 ppm de Zn-Gli; Fase III - 1. dieta + 1600 ppm de ZnO; 2. dieta + 1500 ppm de ZnO + 100 ppm de Zn-Met; 3. dieta + 1500 ppm de ZnO + 100 ppm de Zn-Gli; 4. dieta + 1700 ppm de ZnO; 5. dieta + 1500 ppm de ZnO + 200 ppm de Zn-Met; 6. dieta + 2200 ppm de ZnO+200 ppm de Zn-Gli, em que avaliou-se o desempenho dos animais. Os dados foram analisados utilizando o programa SAS (1998). As médias dos tratamentos foram comparadas pelo teste de Tukey ao nível de 5% de probabilidade, segundo Steel e Torrie (1980). / Given the importance of zinc in the productive and reproductive performance of sows and piglets performance, three experiments to evaluate different sources of zinc in swine were conducted. The first experiment evaluated different sources of zinc, organic (zinc methionine (Zn-Met) and zinc glycine (Zn-Gly)) and inorganic (zinc oxide (ZnO)) in the diet of pregnant gilts and sows lactating and the effects of using these sources in their progeny. Were used 18 female of a commercial strain distributed in experimental design of randomized blocks with three treatments and six repetitions with one animal each. The characteristics evaluated in the sows were: weight, backfat thickness, prolificacy, number of piglets born alive and zinc levels in serum, colostrum and milk; In the piglets after birth until weaning were analyzed: serum levels of zinc, weight gain, bone density and histomorphology enteric. The second experiment 180 piglets were used, weaned at 21 days of age in a randomized block design in a factorial 3 x 3 (3 maternal diets - 100 ppm from ZnO , Zn-Met , Zn-Gly x 3 diets progeny - 100 ppm ZnO , Zn-Met - , Zn-Gly) totaling 9 treatments and 4 replicates of 5 animals each, which evaluated the performance, diarrhea incidence, serum zinc levels, bone density and histomorphology enteric. The third experiment 810 weaned pigs weaned at 21 days of age in a randomized block design with 6 treatments of 3 replicates of 45 animals each: Phases I and II - 1. diet + 2300 ppm of ZnO; 2. diet + 2200 ppm of ZnO + 100 ppm of Zn-Met; 3. diet + 2200 ppm ZnO + 100 ppm of Zn-Gly; 4. diet + 2400 ppm of ZnO; 5. diet + 2200 ppm of ZnO + 200 ppm of Zn- Met; 6. diet + 2200 ppm of ZnO + 200 ppm of Zn-Gly; Phase III - 1. diet + 1600 ppm of ZnO; 2. diet + 1500 ppm of ZnO + 100 ppm of Zn-Met; 3. diet + 1500 ppm of ZnO + 100 ppm of Zn-Gly; 4. diet + 1700 ppm of ZnO; 5. diet + 1500 ppm of ZnO + 200 ppm of Zn- Met; 6. diet + 2200 ppm of ZnO + 200 ppm of Zn-Gly in evaluating the performance of animals. All datas were analyzed using SAS (1998 ) program. The treatment means were compared by Tukey test at 5 % probability , according to Steel and Torrie (1980).

Le diabète de type 2 (DT2) progresse dans le monde entier de manière alarmante. Les complications sont graves et les traitements sont souvent inefficaces et donnent beaucoup d'effets indésirables. Le resvératrol (RSV) et la curcumine (Cur), des polyphénols naturels, sont connus pour leurs différentes activités pharmacologiques, y compris antidiabétiques. Les objectifs de cette étude ont été d'évaluer les effets de la combinaison de ces deux polyphénols sur l'amélioration de la tolérance au glucose. Nos résultats montrent que chez des souris soumises à un régime riche en graisse (HFD), le RSV améliore la tolérance au glucose alors que la Cur n'a pas d'effet significatif. De manière surprenante, lorsque les deux polyphénols sont administrés simultanément, la Cur inhibe l'effet du RSV sur l'amélioration de la tolérance au glucose. Pour expliquer cet antagonisme, nous avons évalué l'effet de ces polyphénols associés ou pas sur le métagénome. Nos résultats indiquent que la Cur et le RSV normalisent la flore intestinale altérée par le HFD. Lors d'une association de ces polyphénols, la Cur inhibe aussi l'effet du RSV sur la croissance de certaines espèces bactériennes. Ainsi, la Cur bloque l'effet inhibiteur du RSV sur Alistipes putredinis. Nos données indiquent que ces polyphénols sont en général anti-inflammatoires mais leur association peut être synergique in vitro ou bien, à l'opposé, antagoniste in vivo pour l'expression de certaines cytokines. Afin d'évaluer l'effet physiologique de la modification de la flore intestinale par les polyphénols, nous avons étudié le métabolome. Parallèlement au métagénome, chacun des polyphénols améliore le profil métabolomique altéré par le HFD. Cependant, la Cur s'oppose à l'effet du RSV. Les données de ce travail indiquent que la Cur n'est pas associable au RSV pour améliorer la tolérance au glucose. / Each year, the worldwide impact of type 2 diabetes (T2D) increases significantly. The complications are serious and treatments are often ineffective and provide a lot of side effects. Resveratrol (RSV) and curcumin (Cur) are natural polyphenols known for their different pharmacological activities, including anti-diabetic. The objectives of this study were to evaluate the effects of the combination of these polyphenols on glucose tolerance improvement. Our results show that in animals fed with a high fat diet (HFD), RSV improved glucose tolerance while Cur displayed no significant effect. Surprisingly, when both polyphenols were administered simultaneously, Cur inhibited the effect of RSV on glucose tolerance improvement. To explain this antagonism between both polyphenols on glucose tolerance, we have evaluated the effect of these compounds on the metagenome associated or not. Our results indicate that RSV and Cur normalized gut flora altered by HFD. When Cur and RSV were administrated simultaneously, Cur inhibited the effect of RSV on the growth of some bacterial species. Thus, Cur blocked the inhibitory effect of RSV on Alistipes putredinis. Our data indicate that these polyphenols displayed generally anti-inflammatory effects, but their combination may be either synergistic in vitro or antagonistic in vivo on the expression of some cytokines. To evaluate the physiological effect of the gut flora changes induced by polyphenols, we have drawn a parallel between metagenome and metabolome data. Each of the polyphenols improved by itself the metabolomic profile altered by HFD. However, Cur hindered the effect of RSV. In a whole, this study indicates that combination of Cur with RSV is not successful to further enhance the RSV-induced improvement of glucose tolerance.

De nos jours, le changement de style de vie et la consommation excessive d'aliments riches en énergie sont associés avec l'augmentation majeure de l'incidence des maladies métaboliques comme l'obésité et le diabète de type 2. Le diabète de type 2 est caractérisé, entre autre, par une augmentation de la production hépatique de glucose responsable d'une hyperglycémie chronique. Durant ces 10 dernières années, plusieurs études ont suggéré que le microbiote intestinal pouvait être impliqué dans le développement des maladies métaboliques. Le microbiote intestinal est composé de plusieurs milliards de bactéries réparties en plus de 1000 espèces différentes qui colonisent le tractus digestif. Plusieurs études ont montré que certaines pathologies comme le diabète et l'obésité sont caractérisées par des altérations taxonomiques et fonctionnelles du microbiote intestinal. De plus, la colonisation de souris axéniques (i.e. dépourvues de microbiote) par un microbiote intestinal provenant de souris ou d'Hommes obèses/diabétiques est suffisante pour induire la pathologie. Ces résultats suggèrent que les modifications du microbiote intestinal retrouvées chez les patients obèses/diabétiques sont potentiellement impliquées dans le développement des maladies métaboliques. Cependant, l'absence de microbiote intestinal chez les souris axéniques induit des altérations structurelles et fonctionnelles de l'intestin comme une hyperperméabilité intestinale ou un système immunitaire atrophié. Dans ces conditions, il est possible de se demander si les effets délétères induits par la colonisation des souris axéniques avec un microbiote modifié peuvent être observés chez des souris conventionnelles. Pour répondre à cette question nous avons développé un nouveau protocole de transfert de microbiote intestinal dans un modèle de souris conventionnelles. Nous avons transféré le microbiote contenu dans le caecum de souris obèses (" microbiote obèse ") et celui contenu dans le caecum de souris minces (" microbiote mince ") dans des souris conventionnelles non traitées aux antibiotiques. De manière surprenante, le transfert du " microbiote obèse " a induit une diminution de la glycémie à jeun associée à une baisse de la néoglucogenèse hépatique chez les souris transplantées. A l'inverse, le transfert du " microbiote mince " n'a pas modifié la néoglucogenèse. De plus, le transfert du " microbiote obèse " a induit des modifications taxonomiques et fonctionnelles du microbiote intestinal des souris transplantées. De manière intéressante, une fois nourries avec un régime hyperlipidique les souris ayant reçu le " microbiote obèse " ont conservé une glycémie à jeun plus faible que les souris non transplantées. Encore une fois, ce phénotype résulte d'une diminution de la production hépatique de glucose caractérisée par une baisse de l'activité des enzymes néoglucogéniques phosphoenolpyruvate carboxykinase et glucose-6-phosphatase. Par ailleurs, ces souris sont également moins grasses que les souris non transplantées. En conclusion, nous avons montré que le transfert d'un " microbiote obèse " peut moduler le métabolisme hépatique et prévenir l'augmentation de la néoglucogenèse hépatique normalement induite par le régime hyperlipidique chez des souris conventionnelles. Ces travaux de thèse ont montré d'une part, que la modification du microbiote intestinal de souris conventionnelles est possible par transfert de microbiote caecal. D'autre part et contre toutes attentes, ces résultats mettent en lumière que, contrairement aux observations faites chez les souris axéniques, le transfert d'un " microbiote obèse " dans une souris conventionnelle n'induit pas les phénotypes caractéristiques des maladies métaboliques. Par ailleurs, ce modèle de transfert caecal pourrait être utile pour la compréhension du rôle des bactéries intestinales sur le développement des maladies métaboliques. / Nowadays, the change of lifestyle and increase in the consumption of high-calorie foods are associated with a marked rise of the prevalence of metabolic diseases, including obesity and type 2 diabetes. Type 2 diabetes is linked, at least in part, to an increase of hepatic glucose production responsible for a fasting hyperglycemia. In the past decade, an increasing body of evidence has proposed gut microbiota as a new factor contributing to these metabolic alterations. Gut microbiota consists of trillions of bacteria identifying more than 1000 different species that inhabit our intestine. A body of work has demonstrated that multiple pathologies such as type 2 diabetes and obesity are characterized by an altered proportion and activity of the gut microbiota. In addition, the colonization of germ-free mice with the gut microbiota from either obese/diabetic humans or obese/diabetic mice transfers the phenotype. These results suggest that the modifications of the gut microbiota found in obese/diabetic patients are a potential etiologic factor for those diseases. Nevertheless, the lack of microbiota in germ-free mice determines both structural and functional alterations such as gut hyperpermeability and the atrophy of the immune system. Therefore, we could wonder whether the detrimental effects of the gut microbiota from obese/diabetic patients observed in germ-free mice may also be observed in healthy conventional mice. To address this issue, we have developed a new gut microbiota transferring process from conventional mice to other mice. We have transferred the cecal microbiota harvested from either obese ("obese microbiota") or lean ("lean microbiota") mice in antibiotic-free conventional mice. Surprisingly, the mice which received the "obese microbiota" had a reduced fasted glycaemia compared to the mice which received the "lean microbiota". This diminution could be attributed to a decrease of the hepatic gluconeogenesis since conversion from pyruvate to glucose and phosphoenolpyruvate carboxykinase activity were lower in the liver of mice which received the "obese microbiota". Conversely, the transfer of the "lean microbiota" did not affect the hepatic gluconeogenesis. In addition, the transfer of the "obese microbiota" changed gut microbiota composition and the microbiome of recipient mice. Interestingly, mice which received the "obese microbiota" and fed a high-fat diet still exhibited reduced fed and fasted glycaemia. Once again, this phenotype was due to a decrease of hepatic gluconeogenesis characterized by a diminution of phosphoenolpyruvate carboxykinase and glucose-6-phosphatase activity. In addition, the mice which received the "obese microbiota" had less adiposity compared to the non-transferred mice. Finally, we reported that transferring the "obese microbiota" impact on hepatic metabolism and prevent HFD-increase hepatic gluconeogenesis. On the one hand, these thesis works, have demonstrated that it is possible to modify the gut microbiota by our caecal transferring process. On the other hand, our results suggest that the transfer of the "obese microbiota" in conventional mice does not induced some characteristics of metabolic diseases contrary to that it is observed in germ-free mice. Furthermore, this kind of gut microbiota transferring process may be useful for a better understanding of the etiology of metabolic diseases.

Acteur clé de la symbiose hôte-microbiote, les IgA sécrétoires modulent le microbiote et participe à l'homéostasie intestinale. Chez la souris, les IgA sont polyréactives, en reconnaissent diverses bactéries, elles régulent la composition du microbiote et réduisent l'inflammation intestinale. Ces observations ouvrent des perspectives thérapeutiques intéressantes. Cependant, les caractéristiques des IgA et leurs spécificités restent mal connues chez l'homme. L'objectif de ce travail a donc été d'étudier la spécificité de la réponse IgA, en distinguant IgA1 et IgA2. La mise au point d'une technique de production in vitro d'IgA monoclonales 100% humaines issues de l'intestin nous a permis de montrer le profil de polyréactivité des IgA. Chaque IgA interagit avec un spectre large mais défini de bactéries commensales. Par ailleurs, les IgA1 et les IgA2 ciblent la même fraction du microbiote. Les réponses IgA1 et IgA2 convergent aussi au niveau des épitopes polysaccharidiques reconnus. Si les IgA sécrétoires contribuent largement à l’homéostasie intestinale, des IgG sériques anti-microbiote jouent un rôle dans la relation symbiotique hôte-microbiote. Leur présence n’ayant pas encore été démontrée chez l’homme en condition physiologique, ce travail a eu pour objectif secondaire de les explorer. Le développement d’une technique de cytométrie bactérienne nous a permis de détecter des IgG ciblant les bactéries commensales dans le sérum des individus sains. Ces IgG anti-microbiote sont dirigées vers les bactéries déjà reconnues par les IgA sécrétoires, elles présentent des spécificités propres à chaque individu. / IgA, the dominant immunoglobulin produced in the gut, plays diverse roles ranging from toxin neutralization, immune functions regulation, intestinal homeostasis maintenance. It is now well established that polyreactive IgA, which target multiple bacteria, can modulate gut microbiota composition and have promising therapeutic effects. However, IgA features remain elusive in humans. We therefore determined the reactivity profile of native monoclonal antibodies from human colon and compared IgA1 and IgA2. We found that IgA are polyreactive and bind a diverse but restricted subset of gut commensals. Most commensals were dually coated by IgA1 and IgA2, yet IgA2 alone coated a distinct fraction of colonic bacteria. Besides their common microbial targets, IgA1 and IgA2 exhibited overlapping anti-carbohydrate repertoires. An essential link between IgA and IgG responses against microbiota has been recently demonstrated in mice. Nevertheless, it remains unclear whether symbiotic bacteria could induce systemic IgG under homeostatic conditions in humans. Hence, we characterized anti-microbiota IgG in serum of healthy donors by bacterial flow cytometry. We found that each individual harbored a diverse and private panel of anti-commensals IgG that converge with secretory IgA to cover a restricted fraction of the gut microbiota. Patients with IgA deficiency or common variable immunodeficiency (CVID) exhibited a distinct set of anti-commensals IgG suggesting that IgA replacement in addition to polyvalent IgG might be beneficial to treat gastro-intestinal symptoms in these patients.