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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Perfil mineral e proteico do plasma seminal de coelhos / Mineral and protein profile of seminal plasma of rabbits

Marco AntÃnio BasÃlio Linard 28 August 2013 (has links)
CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / Seminal plasma is the fluid portion of semen and its presence positively affects the survival and parameters of sperm motility in rabbits. This study aimed (i) to verify the seminal concentrations of sodium, chloride and citric acid as a function of collection month, the color and aspect of the ejaculate, and to find the frequency of ejaculations with the presence of gel fraction, color and aspect of the semen of rabbits on tropical climate, (ii) to meet monthly protein spots and their possible correlation with seminal and biochemical parameters in rabbits. 20 rabbits of New Zealand White, raised in the flat-deck, fed commercial feed were used. The samples were collected twice a week, and then evaluated for volume, color, aspect, vigor, motility and sperm concentration. After the evaluations, the samples were centrifuged to obtain seminal plasma, which was stored in eppendorfs tubes at -18 oC. A monthly seminal plasma pool of each animal was made to evaluate the concentrations of sodium, chloride, citric acid and proteins. Significant monthly variations were found (p <0.05) in the concentrations of sodium, chloride and citric acid in the seminal plasma of rabbits. All mineral constituents analyzed suffered significant influence (p <0.05) by the color of the ejaculate, and the highest concentrations were found in the white-yellowish ejaculates. Correlation studies found a high and significant association between concentrations of sodium and vigor (r = 0.80, p <0.001), and between sperm concentration and citric acid (r = -0.64, p <0.02). Most of the ejaculates of rabbits showed no gel fraction. The results also showed that the white and milky ejaculate are the most common for the species. It was also observed an average concentration of 2.73  0.31 mg / dl total protein in samples of seminal plasma. From the quantification of total protein were prepared two-dimensional electrophoresis gels stained SDS-PAGE with silver nitrate, with a pH gradient between 3 â 10, mesh 15% and a concentration of 100 mg of protein per sample for each month. Gels were analyzed with the Image Master 2D Platinum 6.0.  software. The gel containing the most spots (555 spots) was the gel of the month of May, and the gel with fewer spots (71 spots) was observed in January, but no effect of month on the amount of spots was detected. The majority of spots present in seminal plasma of rabbits have pI below 8, and these spots, about 40% have pI acid, the distribution of the spots as a function of pI was homogeneous throughout the year. The distribution of spots depending on the molecular weight widely varied between the months. Except of a few months, most of the protein had a molecular weight above 100 kDa. The number of protein spot positively and moderately correlated with total protein (r = 0.57, p <0.05) and citric acid (r = 0.59, p <0.05). In silico analysis of 1411 protein spots found compatible proteins with the Swiss-Prote database and TrEMBL (UniProtKB). It is concluded that the composition of the seminal plasma of rabbits showed a broad monthly variation and ejaculated with high concentrations of citric acid are undesirable. In addition, the protein profile of rabbits has great affinity of the proteins to acidic and high molecular weight, no influence of the months of the year on the amount of protein spots identified were found and bioinformatics analysis tool does not provide consistent results with those obtained gel electrophoresis, but it allows an estimate of the probable proteins that can be found. / O plasma seminal representa a porÃÃo fluida do sÃmen e sua presenÃa afeta positivamente a sobrevivÃncia e os parÃmetros de motilidade espermÃtica em coelhos. Este trabalho teve como objetivos (i) verificar as concentraÃÃes seminais de sÃdio, cloretos e Ãcido cÃtrico em funÃÃo do mÃs de coleta, da cor e do aspecto do ejaculado, e encontrar a frequÃncia de ejaculados com presenÃa de fraÃÃo gel, cor e aspecto do sÃmen de coelhos criados em clima tropical; (ii) conhecer os spots proteicos mensais e suas possÃveis correlaÃÃes com parÃmetros seminais e bioquÃmicos de coelhos. Foram utilizados 20 coelhos da raÃa Nova ZelÃndia Branca, criados em sistema flat-deck, alimentados com raÃÃo comercial. Os ejaculados foram coletados duas vezes por semana, e logo avaliados quanto ao volume, cor, aspecto, vigor, motilidade e concentraÃÃo espermÃtica. ApÃs as avaliaÃÃes os ejaculados foram centrifugados para obtenÃÃo do plasma seminal, que foi acondicionado em tubos eppendorfs a -18 oC. Um pool mensal de PS de cada animal foi feito para avaliaÃÃo das concentraÃÃes de sÃdio, cloretos, Ãcido cÃtrico e proteÃnas totais. Foram constatadas variaÃÃes mensais significativas (p<0,05) nas concentraÃÃes de sÃdio, cloretos e Ãcido cÃtrico no plasma seminal de coelhos. Todos os constituintes minerais analisados sofreram influencia significativa (p<0,05) da cor do ejaculado, e as concentraÃÃes mais elevadas foram constatadas nos ejaculados de cor branco-amarelada. O estudo de correlaÃÃes encontrou associaÃÃo alta e significativa entre as concentraÃÃes de sÃdio e vigor (r=0,80; p<0,001), bem como entre a concentraÃÃo espermÃtica e o Ãcido cÃtrico (r=-0,64; p<0,02). A maior parte dos ejaculados de coelhos nÃo apresentaram a fraÃÃo gel. Os resultados tambÃm mostraram que o ejaculado de cor branca e de aspecto leitoso sÃo os mais comuns nessa espÃcie. TambÃm foi verificado uma concentraÃÃo mÃdia de 2,73 Â0,31 &#956;g/dl de proteÃnas totais nas amostras de plasma seminal. A partir da quantificaÃÃo das proteÃnas totais foram confeccionados dois gÃis de eletroforese bidimensional SDS-PAGE corados com nitrato de prata, com gradiente de pH entre 3 â 10, malha de 15% e uma concentraÃÃo de 100 &#956;g de proteÃnas por amostra, para cada mÃs. Os gÃis foram analisados no software Image Master 2D Platinum 6.0. Â. O gel que continha o maior nÃmero de spots (555 spots) foi o do mÃs de maio, e o gel com menor nÃmero spots (71 spots) foi verificado em janeiro, mas nÃo houve efeito do mÃs sobre a quantidade de spots detectados. A maioria dos spots presentes no plasma seminal de coelhos tem pI abaixo de 8 e, destes spots, cerca de 40% tem pI Ãcido, a distribuiÃÃo dos spots em funÃÃo do pI foi homogÃnea ao longo do ano. A distribuiÃÃo dos spots em funÃÃo do peso molecular variou amplamente entre os meses. Com exceÃÃo de alguns os meses, a maioria das proteÃnas apresentaram peso molecular acima de 100 kDa. O nÃmero de spot proteicos correlacionaram-se moderada e positivamente com as proteÃnas totais (r=0,57; p<0,05) e com o Ãcido cÃtrico (r=0,59; p<0,05). A anÃlise in silico dos spots encontrou 1.411 proteÃnas compatÃveis com os bancos de dados Swiss-Prote e TrEMBL (UniProtKB). Concluiu-se que a composiÃÃo do plasma seminal de coelhos apresentou ampla variaÃÃo mensal e que ejaculados com alta concentraÃÃo de Ãcido cÃtrico sÃo indesejÃveis. AlÃm disso, o perfil proteico de coelhos apresenta grande parte das proteÃnas com afinidade a meio Ãcido e com alto peso molecular, nÃo houve influencia de meses do ano sobre a quantidade de spots proteicos detectados, e devido à ausÃncia de dados sobre coelhos, a ferramenta de anÃlise bioinformÃtica nÃo ofereceu resultados coerentes, mas permite uma estimativa das provÃveis proteÃnas que podem ser encontradas.

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