Cryopreservation of epididymal spermatozoa may be the only opportunity to preserve valuable genetics of males in cases of unforeseen injury or death. Stallion epididymal spermatozoa have been cryopreserved successfully and it has been demonstrated that stallion epididymal spermatozoa are fertile, and pregnancies as well as live foals have been produced. As spermatozoal quality parameters like motility, morphology and viability have a major influence on fertility and pregnancy rates, it is of great interest to describe these and investigate the influence of seminal plasma on these parameters. Fresh and post-thaw parameters (motility, morphology and viability) of stallion epididymal spermatozoa that have been and that have not been exposed to seminal plasma were evaluated, and directly compared to fresh and post-thaw parameters of ejaculated spermatozoa from the same stallions. Six sperm categories of each stallion (n= 4) were evaluated for motility, morphology and viability. These categories were fresh ejaculated spermatozoa (Fr-E), fresh epididymal spermatozoa that had been exposed to seminal plasma (Fr-SP+), fresh epididymal spermatozoa that had never been exposed to seminal plasma (Fr-SP-), frozen-thawed ejaculated spermatozoa (Cr-E), frozen-thawed epididymal spermatozoa that had been exposed to seminal plasma prior to freezing (Cr-SP+) and frozen-thawed epididymal spermatozoa that had never been exposed to seminal plasma (Cr-SP-). Results show that seminal plasma stimulates initial motility of fresh epididymal stallion spermatozoa while this difference in progressive motility is no longer present post-thaw; and that progressive motility of fresh or frozen-thawed ejaculated stallion spermatozoa is not always a good indicator for post-thaw progressive motility of epididymal spermatozoa. This study shows that seminal plasma has a positive influence on the incidence of overall sperm defects, midpiece reflexes and distal cytoplasmic droplets in frozen-thawed stallion epididymal spermatozoa while the occurance of midpiece reflexes is likely to be linked to distal cytoplasmic droplets. Furthermore, we could show that seminal plasma does not have an influence on viability of fresh and frozen-thawed morphologically normal epididymal spermatozoa. In conclusion, we recommend the retrograde flushing technique using seminal plasma as flushing medium to harvest and freeze stallion epididymal spermatozoa. AFRIKAANS : Bevriesing van epididimale spermatozoa mag die enigste geleentheid wees om waardevolle genetika van manlike diere te bewaar in die geval van onverwagse dood of besering. Epididimale spermatozoa van hingste is al suksesvol bevries, is bewys vrugbaar te wees en dragtighede sowel as lewendige vullens is deur die gebruik daarvan gelewer. Aangesien maatstawwe van spermgehalte soos beweeglikheid, morfologie en lewensvatbaarheid ‘n beduidende invloed uitoefen op vrugbaarheid en dragtigheidsyfers, is dit van groot belang om hierdie maatstawwe te omskryf en die invloed van seminale plasma daarop te ondersoek. Vars en ontdooide maatstawwe (beweeglikheid, morfologie en lewensvatbaarheid) van hings epididimale spermatozoa wat of aan seminal plasma blootgestel is of nie, is vergelyk met vars en ontdooide maatstawwe van geejakuleerde spermatozoa. Ses spermkategorieë van elke hings (n=4) is geevalueer vir beweeglikheid, morfologie en lewensvatbaarheid. Die kategorieë was vars geejakuleerde spermatozoa (Fr-E), vars epididimale spermatozoa wat blootgestel is aan seminale plasma (Fr-SP+), vars epididimale spermatozoa sonder blootstelling aan seminale plasma (Fr-SP-), ontdooide geejakuleerde spermatozoa (Cr-E), ontdooide epididimale spermatozoa blootgestel aan seminale plasma voor bevriesing (Cr-SP+) en ontdooide epididimale spermatozoa wat nooit blootgestel is aan seminale plasma nie (Cr-SP-). Uitslae toon dat seminale plasma die aanvanklike beweeglikheid van vars epididimale hings spermatozoa stimuleer, terwyl die verskil in lynbeweeglikheid nie meer teenwoordig is na ontdooiing nie; dat lynbeweeglikheid van vars epididimale spermatozoa of ontdooide geejakuleerde hings spermatozoa nie altyd ‘n goeie aanduiding is vir lynbeweeglikheid na ontdooiing nie. Hierdie studie toon dat seminale plasma ‘n positiewe invloed het op die voorkoms van algehele spermdefekte, midstukreflekse en distale sitoplasmiese druppeltjies in ontdooide hings epididimale spermatozoa terwyl die voorkoms van midstuk reflekse waarskynlik verband hou met distale sitoplasmiese druppletjies. Verder kon ons ook aantoon dat seminale plasma geen invloed het op die lewensvatbaarheid van vars en ontdooide morfologies normale epididimale spermatozoa nie. Ten slotte beveel ons aan dat retrograadse spoeling met seminale plasma as spoelmedium gebruik word wanneer hings epididimale spermatozoa versamel en bevries word. / Dissertation (MSc)--University of Pretoria, 2010. / Production Animal Studies / unrestricted

SILVA, Katiane Queiroz da. Efeito da adição de plasma seminal e de sua composição bioquimica sobre os espermatozóides epididimários de caprinos. 2009. 43 f. : Dissertação (mestrado) - Universidade Federal do Ceará, Centro de Ciências Agrárias, Departamento de Zootecnia, Fortaleza-CE, 2009 / Submitted by Nádja Goes (nmoraissoares@gmail.com) on 2016-08-02T14:51:18Z No. of bitstreams: 1 2009_dis_kqsilva.pdf: 313426 bytes, checksum: 9fae2725277c1fe6e2b27d9bb34c0908 (MD5) / Approved for entry into archive by Nádja Goes (nmoraissoares@gmail.com) on 2016-08-02T14:51:56Z (GMT) No. of bitstreams: 1 2009_dis_kqsilva.pdf: 313426 bytes, checksum: 9fae2725277c1fe6e2b27d9bb34c0908 (MD5) / Made available in DSpace on 2016-08-02T14:51:57Z (GMT). No. of bitstreams: 1 2009_dis_kqsilva.pdf: 313426 bytes, checksum: 9fae2725277c1fe6e2b27d9bb34c0908 (MD5) Previous issue date: 2009 / The objective of this study was to verify the effect of the addition and of the composition of the seminal plasma (SP) on the characteristics of the epididymal spermatozoa (ES) of goat. Eight goat males were used for obtaining of the biological material so that, the SP it was obtained in previous collections and stored at -18 oC until that proceeded to the addition to ES and the biochemical analyses. ES were obtained of the tail of the epididymal by the method of surgical castration. Four sample of 100 μL were removed from the "pool" of ES, and in two of them 120 μL were added of SP. Two sample, one with and other without SP, were diluted in a concentration of 200 x 106 sptz /mL, in the extender citrate-yolk (CY) and in the extender tris-yolk (TY). The samples were evaluated the vigor, motility and motility degradation rate (TDM) in fresh state and for the test of slow thermoresistence, after two and 24 hours of conservation. For the determination of fructose levels and total proteins were used specific kits of In Vitro Diagnóstico S/A®. The determination of activity of the phospholipase A2 was realized by the technique of the pH-stat. For the analysis of the data was used the statistical program SAS@. The results demonstrated that the addition of SP to ES reduced seminal parameters significantly, except in the extender TY when the animals were to grouped in function of concentration of total proteins of SP. Besides, the TY, when used to conserve the ES added of SP with low fructose concentration (540 mg/dL) preserved the seminal characteristics better. It follows that the initial concentration of fructose in the seminal plasma influenced the quality of conservation of the epididymal spermatozoa; as for the total proteins just affected the conservation in the citrate-yolk and the intensity of activity of the phospholipase A2 did not interfere in the conservation at 5 ºC of the epididymal spermatozoa of goat / O objetivo deste estudo foi verificar o efeito da adição e da composição do plasma seminal (PS) sobre as características dos espermatozóides epididimários (EEP) de caprinos. Utilizaram-se oito machos caprinos para obtenção do PS, que foi obtido em coletas prévias e armazenado a -18o C até que se procedessem a sua adição aos EEP e as análises bioquímicas. Os EEP foram obtidos da cauda do epidídimo pelo método de castração cirúrgica. Do “pool” de EEP foram retiradas quatro alíquotas de 100 μL, sendo que em duas delas foram adicionados 120 μL de PS e nas outras duas não. Duas alíquotas, uma com e outra sem PS, foram diluídas a uma concentração de 200 x 106 sptz/ mL, no diluidor citrato-gema (CG) e no diluidor tris-gema (TG). As amostras foram avaliadas quanto ao vigor, à motilidade e à taxa de degradação da motilidade (TDM) no seu estado fresco e pelo teste de termorresistência lento, após duas e 24 horas de conservação. Para a determinação dos níveis de frutose e de proteínas totais foram utilizados os kits específicos da In Vitro Diagnóstico S/A®. A determinação da atividade da fosfolipase A2 foi realizada pela técnica do pH-stat. Para a análise dos dados foi utilizado o programa estatístico SAS@. Os resultados demonstraram que a adição de PS aos EEP diminuiu significativamente os parâmetros seminais avaliados, exceto no diluidor TG quando os animais foram agrupados em função da concentração de proteínas totais do PS. Além disso, o TG, quando utilizado para conservar os EEP adicionados de PS de baixa concentração de frutose (540 mg/dL) preservou melhor as características seminais avaliadas. Concluiu-se que a concentração inicial de frutose no PS influenciou positivamente a qualidade de conservação dos espermatozóides epididimários; já a de proteínas totais afetou apenas a conservação no CG e a intensidade de atividade da fosfolipase A2 não interferiu na conservação a 5 ºC dos espermatozóides epididimários de caprinos

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / O objetivo deste estudo foi verificar o efeito da adiÃÃo e da composiÃÃo do plasma seminal (PS) sobre as caracterÃsticas dos espermatozÃides epididimÃrios (EEP) de caprinos. Utilizaram-se oito machos caprinos para obtenÃÃo do PS, que foi obtido em coletas prÃvias e armazenado a -18o C atà que se procedessem a sua adiÃÃo aos EEP e as anÃlises bioquÃmicas. Os EEP foram obtidos da cauda do epidÃdimo pelo mÃtodo de castraÃÃo cirÃrgica. Do âpoolâ de EEP foram retiradas quatro alÃquotas de 100 μL, sendo que em duas delas foram adicionados 120 μL de PS e nas outras duas nÃo. Duas alÃquotas, uma com e outra sem PS, foram diluÃdas a uma concentraÃÃo de 200 x 106 sptz/ mL, no diluidor citrato-gema (CG) e no diluidor tris-gema (TG). As amostras foram avaliadas quanto ao vigor, à motilidade e à taxa de degradaÃÃo da motilidade (TDM) no seu estado fresco e pelo teste de termorresistÃncia lento, apÃs duas e 24 horas de conservaÃÃo. Para a determinaÃÃo dos nÃveis de frutose e de proteÃnas totais foram utilizados os kits especÃficos da In Vitro DiagnÃstico S/AÂ. A determinaÃÃo da atividade da fosfolipase A2 foi realizada pela tÃcnica do pH-stat. Para a anÃlise dos dados foi utilizado o programa estatÃstico SAS@. Os resultados demonstraram que a adiÃÃo de PS aos EEP diminuiu significativamente os parÃmetros seminais avaliados, exceto no diluidor TG quando os animais foram agrupados em funÃÃo da concentraÃÃo de proteÃnas totais do PS. AlÃm disso, o TG, quando utilizado para conservar os EEP adicionados de PS de baixa concentraÃÃo de frutose (540 mg/dL) preservou melhor as caracterÃsticas seminais avaliadas. Concluiu-se que a concentraÃÃo inicial de frutose no PS influenciou positivamente a qualidade de conservaÃÃo dos espermatozÃides epididimÃrios; jà a de proteÃnas totais afetou apenas a conservaÃÃo no CG e a intensidade de atividade da fosfolipase A2 nÃo interferiu na conservaÃÃo a 5 ÂC dos espermatozÃides epididimÃrios de caprinos / The objective of this study was to verify the effect of the addition and of the composition of the seminal plasma (SP) on the characteristics of the epididymal spermatozoa (ES) of goat. Eight goat males were used for obtaining of the biological material so that, the SP it was obtained in previous collections and stored at -18 oC until that proceeded to the addition to ES and the biochemical analyses. ES were obtained of the tail of the epididymal by the method of surgical castration. Four sample of 100 μL were removed from the "pool" of ES, and in two of them 120 μL were added of SP. Two sample, one with and other without SP, were diluted in a concentration of 200 x 106 sptz /mL, in the extender citrate-yolk (CY) and in the extender tris-yolk (TY). The samples were evaluated the vigor, motility and motility degradation rate (TDM) in fresh state and for the test of slow thermoresistence, after two and 24 hours of conservation. For the determination of fructose levels and total proteins were used specific kits of In Vitro DiagnÃstico S/AÂ. The determination of activity of the phospholipase A2 was realized by the technique of the pH-stat. For the analysis of the data was used the statistical program SAS@. The results demonstrated that the addition of SP to ES reduced seminal parameters significantly, except in the extender TY when the animals were to grouped in function of concentration of total proteins of SP. Besides, the TY, when used to conserve the ES added of SP with low fructose concentration (540 mg/dL) preserved the seminal characteristics better. It follows that the initial concentration of fructose in the seminal plasma influenced the quality of conservation of the epididymal spermatozoa; as for the total proteins just affected the conservation in the citrate-yolk and the intensity of activity of the phospholipase A2 did not interfere in the conservation at 5 ÂC of the epididymal spermatozoa of goat

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