Orientador: Nereu Carlos Prestes / Resumo: A proposta do presente estudo foi descrever as diferenças no padrão morfológico das células do fluido amniótico, no momento do parto. Comparando os resultados obtidos de bezerros nelores oriundos de produção in vitro, transferência de embriões e inseminação artificial. Utilizaram-se 60 animais, divididos em Grupo 01: vinte vacas inovuladas com embriões Nelore produzidos in vitro (PIV) a partir de colheita de oócitos por aspiração folicular de doadoras, Grupo 02: vinte vacas gestando bezerros nelores originados de inovulações de embriões oriundos de doadoras superovuladas pelo método convencional, e Grupo 03: vinte vacas nelores PO gestando bezerros oriundos de inseminação artificial (controle). Próximo ao parto as vacas foram transferidas para piquete maternidade com acompanhamento dos partos. Durante a fase de expulsão e após a ruptura do alantocórion realizou-se a punção do âmnion para colheita de 20mL de liquido, que foi depositado em tubo plástico e congelado em freezer. Para a classificação celular as amostras, pós-descongelação, foram centrifugadas na Citocentrifuga (Revan Centrifuga Ciclo Cito) a 4000rpm durante seis minutos e empregou-se a técnica de coloração Hematoxilina-Shorr. Ao final deste procedimento, as lâminas citológicas foram montadas com lamínulas empregando Bálsamo do Canadá e posteriormente (após o período de secagem) observadas em microscópio óptico com aumentos de 200x e 400x para visualização das características morfológicas e tintoriais das células epiteliais do feto. Para avaliação da maturidade epidérmica utilizou-se a técnica de coloração de Sulfato Azul do Nilo bem como a porcentagem de células queratinizadas (coloração Hematoxilina-Shorr). As células basais e parabasais não foram encontradas nas lâminas citológicas das amostras de líquido amniótico analisadas... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The aim of the present study was to describe the differences in the cytological pattern of the amniotic fluid, at the moment of delivery, comparing the data from Nelore calves obtained by in vitro production, embryo transfer and artificial insemination. Sixty (n=60) Nelore cows were divided in group 01: Twenty cows (n=20) pregnant with Nelore calves obtained by in vitro production (IVP) after follicular aspiration; group 02: Twenty cows (n=20) pregnant with Nelore calves obtained by superovulation of embryo donors (ET); group 03: Twenty Nelore cows (n=20) pregnant of calves obtained by artificial insemination (AI; control). Near to the labor, the cows were transferred to a maternal paddock, permitting delivery observation. During the expulsion phase, after the rupture of the alantochorion the amnion was punctured and 20ml of fluid were collected in a plastic device and frozen. The samples, after being thawed, were centrifuged at 4000 rpm, during 06 minutes the Hematoxilin-Shorr stain was applied, after this, the glass slides were covered with Canadian balsame and analyzed in optical microscopy. To the evaluation of the epidermal maturity were used the Blue Nile Sulfate stain, as well the percent of keratinizaded cells observed in the Hematoxilin-Shorr stain. The basal and parabasal cells were not found on the slides of the amniotic fluid samples analyzed. The small intermediate cells (CIP) had its shape as an oval or polygonal form with large cytoplasm and central nucleus. The large intermediate cell (CIG) showed a central nucleus and a bigger nucleus-cytoplasm relation compared to the superficial cells. These were the largest cells found. They showed keratinization with angular edges, having both a picnotic nucleus (CSN) and no nucleus at all. (CSA). The technique of Blue Nile Sulfate stain did not showed satisfactory for the evaluation of the fetal maturity, in spite of some records... (Complete abstract click electronic access below) / Mestre

Orientador: Márcia Guimarães da Silva / Banca: Rodrigo Paupéro de Camargo Soares / Banca: Vera Lúcia Mores Rall / Não disponível / Abstract: Microbial invasion of the amniotic cavity has been described in term deliveries and its role on the immune modulation is of interest to the better understanding of the underlying labor processes. The aim of this study was to determine the prevalence of Mycoplasma hominis and Ureaplasma urealyticum in the amniotic fluid of term pregnancies and to evaluate its influence on cytokines production at the end of pregnancy. A cross sectional study was conducted with fifty five pregnant women out of labor with intact membranes and gestational age between 37 and 41 weeks seen at the Bom Jesus hospital in Ariquemes, Rondônia, between June 2009 and May 2010. Amniotic fluid samples and fragments of chorioamniotic membranes were collected at cesarean section. M. hominis and U. urealyticum detection was performed by PCR and Interleukin (IL)-1β, IL-6, IL-8, IL-10 and Tumor Necrosis Factor (TNF)- levels were determined by ELISA. Chorioamniotic membranes were submitted to histopatological analyses. Presence of M. hominis was detected in 36.4% of amniotic fluid samples and any of them was positive for U. urealyticum. Regarding cytokines levels, 63.6% and 90.9% of samples have not shown detectable concentrations of TNF- and IL-1β. The median concentration of IL-6 and IL-8 were 107.9 pg/mL (0-517.1) and 208.1 pg/mL (0-1897.4), respectively. Interleukin-1, IL-6, IL-8 and TNF- concentrations were not associated with the presence of M. hominis in amniotic fluid, regardless the gestational age. No sample had detectable IL-10 levels. The histopatological analyses have shown no chorioamnionitis in any of the membranes, only a discreet mononuclear infiltration in the decidua could be observed in 40.4% of the samples. Presence of M. hominis was detected in 36.4% of amniotic fluid samples and any of them was positive for U. urealyticum. Regarding cytokines levels, 63.6% and 90.9% of samples have not... (Complete abstact click electronic access below) / Mestre

Orientador: Nereu Carlos Prestes / Banca: Maria Denise Lopes / Banca: João Carlos Pinheiro Ferreira / Banca: Márcia Guimarães da Silva / Banca: Daniel Bartole de Sousa / Resumo: Tendo em vista, a incidência de problemas fetais observados no momento do parto de animais produzidos in vitro (PIV) e a escassez de literatura nesta área, o presente estudo teve por objetivo determinar os níveis de fator de necrose tumoral-alfa (TNF-), interferon-gama (IFN-), interleucina-6 (IL-6) e IL-8 no fluido amniótico de bezerros oriundos de PIV comparando com aqueles obtidos pela transferência de embrião convencional e inseminação artificial no momento do parto, na tentativa de auxiliar na resolução de problemas ainda pouco conhecidos que são observados a campo durante emprego comercial da PIV. Foram utilizados sessenta animais divididos em grupo 01: vinte vacas gestando bezerros da raça Nelore PO de inovulações de embriões oriundos de doadoras superovuladas pelo método convencional (TE); grupo 02: vinte vacas gestando bezerros Nelore PO de inovulações de embriões oriundos de PIV; grupo 03: vinte vacas Nelore PO gestando bezerros oriundos de inseminação artificial (IA) - controle. Todos os partos dos animais do experimento foram observados. Durante a fase de expulsão e após a ruptura do alantocórion, foi realizada a punção do âmnion para a colheita de 15mL de líquido amniótico utilizando-se agulhas e seringas descartáveis compatíveis. O material foi depositado em tubo plástico e congelado em freezer para posterior análise laboratorial. O acompanhamento do padrão de desenvolvimento dos bezerros no periparto foi realizado. Após a descongelação das amostras foi realizado ensaio imunoenzimático (ELISA) para a dosagem de TNF-, com emprego do Kit comercial: ESS0011 - Bovine TNF Alpha, para IFN- foi Kit comercial: ESS0026 - Bovine IFN-, para IL-6 foi Kit comercial: ESS0029 - Bovine IL-6 da Pierce Biotechnology e para IL-8 Kit... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: In view, the incidence of fetal problems seen at the time of birth of animals in vitro production and the be short of the literature in this area, this study aimed to determine the levels of tumor necrosis factor (TNF-), interferon-gamma (IFN-), interleukin-6 (IL-6) and IL-8 in amniotic fluid of calves from in vitro production (IVP) compared with those obtained by conventional embryo transfer and artificial insemination at the delivery, in an attempt to assist in solving problems that are still little known observed in simulations and the experimental field of commercial employment during IVP. Sixty animals used were divided into groups: 1 - Twenty Nelore cross-breed cows pregnant with Nelore calves of embryos transfer conventional method (ET); 2 - Twenty Nelore cross-breed cows pregnant with Nelore calves obtained by in vitro production after follicular aspiration; 3 - Twenty pregnant Nelore cows bearing calves obtained by artificial insemination (AI). All births of the animals of the experiment were observed. During the expulsion phase the amnion was punctured and 15mL of fluid were collected using a needle and syringes compatible. The material was deposited on plastic tube and frozen in freezer for later analysis. Monitoring the pattern of development of the calves in peripartum was performed. The cytokines levels were measured by immunoenzymatic assay (ELISA). The commercial kit used was: ESS0011 - Bovine TNF Alpha, ESS0026 - Bovine IFN- gamma, ESS0029 - Bovine IL-6 Pierce Biotechnology and comercial Kit: D8000C - Human IL-8 of the R & D Systems. The protocol was performed according to the manufacturer's recommendations. The densities were evaluated in automatic optical reader ELISA. The well plates were read at 450nm. The Kruskal-Wallis Test was used for statistical analysis... (Complete abstract click electronic access below) / Mestre