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Made available in DSpace on 2014-06-11T19:35:12Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-12-09Bitstream added on 2014-06-13T18:46:36Z : No. of bitstreams: 1 martins_lr_dr_botfmvz.pdf: 1331957 bytes, checksum: 49bb483ccab8a8fb76bb4bcfe96d230a (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Testou-se a hipótese de influência da sazonalidade sobre a produção embrionária in vitro de gato doméstico (Felis catus), na cidade de Botucatu, localizada sob latitude de 22º53’09” S, no estado de São Paulo, Brasil, em associação a três condições ovarianas – inativa, luteal e folicular em dois momentos: período I, que compreendeu os meses de janeiro, fevereiro e março e período II constituído pelos meses de agosto, setembro e outubro. Realizaram-se dois experimentos: I) Aspectos relacionados à maturação in vitro e análise ultra-estrutural de complexos cumulus-oophorus (COC) de felinos domésticos e II) Produção in vitro e análise ultra-estrutural de embriões de gato doméstico. O objetivo do experimento I foi avaliar as taxas de recuperação e maturação in vitro de COC e a descrição de suas características ultra-estruturais. O experimento II visou à avaliação das taxas de clivagem, de formação de mórulas, de produção de blastocistos; à avaliação da qualidade dos embriões felinos em fase inicial de desenvolvimento e à descrição das características ultra-estruturais dos embriões obtidos. No experimento I, para verificação das taxas de recuperação oocitária e de maturação in vitro foram avaliados 473 oócitos grau I no período I e 482 oócitos grau I no período II, provenientes de 20 e de 10 pares de ovário, respectivamente. O número médio de oócitos obtidos por gata foi de 23,65 e 48,2 nos períodos I e II, respectivamente. Esses COC foram maturados in vitro e classificados quanto à configuração nuclear em vesícula germinativa (VG), quebra de vesícula germinativa (QVG), metáfase I (MI), metáfase II (MII) e degenerados/não identificados (D/NI). No experimento II, 386 e 568 COC durante o período I e II respectivamente, foram fertilizados com sêmen fresco e após três e oito dias de cultivo verificou-se... / The hypothesis of season effect at Botucatu, 22º53’09” S, Sao Paulo State, Brazil, on in vitro embryo production of domestic cat (Felis catus), was tested in association with three ovarian conditions – inactive, luteal and follicular during two seasons: season I, which comprehended the months of January, February and March and season II, constituted by the months of August, September and October. This study was divided in two experiments: I) In vitro oocyte maturation and ultrastructural analysis of cumulus-oocyte complexes (COC) of domestic cats and II) In vitro embryo production and ultrastructural analysis of embryo of domestic cats. The aim of the experiment I was to analyze the COC recovery rate and in vitro nuclear maturation (IVM) rate and the description of its ultrastructural characteristics. The experiment II aimed to evaluate cleavage and morulae rates and blastocyst formation; to evaluate the quality of domestic cat embryos and to describe ultrastructural characteristics of these embryos. In experiment I, to evaluate COC recovery rate and in vitro maturation rate; 473 Grade I COC recovered from 20 ovarian pairs and 482 Grade I COC recovered from 10 ovarian pairs were evaluated during season I and season II, respectively. These COC were in vitro matured and classified according to nuclear configuration as germinal vesicle (GV), germinal vesicle breakdown (GVBD), metaphase I (MI), metaphase II (MII) and degenerated/non-identified (D/NI). In experiment II, 368 and 568 COC recovered from ovaries during season I and II, respectively, were in vitro fertilized with fresh semen and after three and eight days of culture the development stage and embryo viability were analyzed. In experiment I, no significant difference was verified in the percentage of oocytes in MII in both seasons and considering the three conditions. These results indicate that in vitro maturatiom... (Complete abstract click electronic access below)

Orientador: Sergio Ricardo Batlouni / Banca: Maria Inês Borella / Banca: Carlos Alberto Vicentini / Resumo: A reprodução de peixes migradores nativos apesar de já apresentar relativo sucesso, ainda apresenta diversos obstáculos. Entre eles podemos destacar as variações nas taxas de fertilidade, eclosão e sobrevivência de larvas entre desovas. Desta forma, o objetivo deste trabalho foi avaliar de forma inédita se o manejo pode interferir neste processo em machos de P. lineatus. Os resultados mostraram que os machos mantidos em tanques-rede (TR) com água de baixa qualidade durante 18 meses apresentam encurtamento do ciclo reprodutivo; redução significativa no IGS e no diâmetro de túbulos seminíferos (p<0,05) quando comparados com machos mantidos em viveiros escavados (VE). A concentração espermática e as taxas de fertilidade obtidas com o sêmen dos machos de TR foram inferiores às obtidas com o sêmen dos machos em VE (p<0,05). Além disso, os espermatozoides de TR apresentaram maior frequência de danos de DNA, mesmo apresentando taxas similares de motilidade e sobrevivência que os de VE (p<0,05). Os resultados mostraram de forma inédita que o manejo inadequado pode interferir na gametogênese e na qualidade de gametas masculinos nesta espécie. Além disso, demonstramos que práticas inadequadas de manejo podem prejudicar a função testicular ainda nos meses de inverno, muito antes dos meses de reprodução / Abstract: The reproduction of native migratory fish still presents several obstacles. Among them we can highlight the changes in fertility rates, hatching and survival of larvae among spawning. Thus, the objective of this study was to evaluate in an unprecedented way if the reproductive management can interfere in this process in males of P. lineatus. The results showed that males kept in cages (C) for 18 months have their reproductive cycle shortened; significant reduction in the GSI and in the diameter of seminiferous tubules (p<0.05) compared with males kept in earths ponds (EP). The sperm concentration and fertility rates obtained with semen from males of Cs were lower than those obtained with semen EP (p<0.05). In addition, the sperm of males maintained in Cs had higher frequency of DNA damage, even with similar rates of survival and motility of the EPs (p<0.05). The results showed by unprecedented manner that the improper management can interfere with gametogenesis and quality of male gametes in this species. Furthermore, we demonstrate inadequate management practices can affect the testicular development even in the winter months, long before the breeding season / Mestre

Improvements in turkey reproductive efficiency is a very desirable goal for the turkey industry. The ability to maintain turkey hens in the egg-laying (LAY) stage and produce one additional egg per hen a year is estimated to save the turkey industry approximately $1.5 million dollars per year. Overall protein expression generated by tissues of the hypothalamic-hypophyseal complex, namely the anterior pituitary, of the mature turkey hen have a profound impact on reproductive cycling (Scanes, 2000). One of the key physiological factors produced by the anterior pituitary and shown to play a significant role in the regulation of egg laying is the protein prolactin (Prl). The objectives for this study are to examine the overall protein expression patterns from turkey hen pituitary tissue during the nonphotostimulated (NPS), photostimulated (PS), and egg laying (LAY) stages. Attempts to isolate transcription factors that regulate the expression of Prl using an affinity chromatography technique or southwestern screening of a bacteriophage expression library were not successful. A global analysis of protein expression, using two-dimensional polyacrylamide gels (2D gels), was conducted using whole cell, cytoplasmic and nuclear protein extracts from pituitary tissue collected during the NPS, PS and LAY reproductive stages. Approximately 1,046 proteins ranging in pI from 4.6-8.2 and molecular weights between 100 kDa-6kDa were resolved. Protein expression patterns were replicated and verified using pituitaries harvested from NPS, PS and LAY stage turkey hens from another laboratory. Proteins showing considerable changes (563 proteins increased in expression and 98 proteins decreased in expression from the NPS to the LAY stage) in their expression between the reproductive stages were grouped in analysis sets for future identification. These proteins may prove to be important to the reproductive cycling of the turkey hen and warrant future investigation. The results of this study contribute to the overall understanding of the role that the pituitary, as a critical part of the hypothalamic-hypophyseal complex, plays in turkey hen reproductive cycling. / Master of Science