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Produ??o in vivo e identifica??o do sexo de embri?es h?bridos Equus caballus X Equus asinus / In vivo production and sex determination of hybrid embryos Equus caballus x Equus asinus

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Previous issue date: 2015-11-06 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Embryo transfer and other biotechnologies are intensivelyused for equine reproduction and other species as well. Even though there is an expansion of the mule market and an increase in the number of animals, researches working with reproduction of these animals are still scarce. The aim of this study was to evaluate aspects of embryo transfer, embryo morphology and gender identification of hybrid (horses x donkeys) embryos, checking either their similarity or divergence with the characteristics already known of equine embryos. Attempts of embryo collection on Day 6 -9 after OV were performed in mares previously bred with a P?ga donkey. The embryo recovery rates, the characteristics related to age, morphology and embryonic diameter were evaluated. After these assessments, a group of embryos was cut using an adapted technique, where, the cutting process was carried out with the aid of an ophthalmic scalpel blade. The resulting biggest cut portion was used for embryo transfer, and the smallest parts and the others whole embryos to determination of the embryo gender. Data were analyzed using Fisher's exact test, with 5% significance, except the daily growth of embryos, which was analyzed by linear regression. The overall embryo recovery rate was 55,9% (71/127), and for each group D6, D7, D8 and D9, was 57,1% (28/49), 51% (25/49), 63% (15/24) 60% (3/5), respectively. The developmental stage of the collected embryos were, morula: 18.3% (13/71); early blastocysts: 26,8% (19/71); blastocyst: 29,6% (21/71); and expanded blastocysts: 25,3% (18/71). The measured diameter revealed that the size of the embryos ranged from 147 to 1688?m, the mean diameter of all collected embryos was 438,04?m, and according to different groups, the average size (smallest and largest diameter) of embryos were D6 (n = 29) - 183,4?m (147 - 253?m), D7 (n = 24) - 463,2?m (168 - 886?m), D8 (n = 15) - 727,2?m (422 - 1224?m ) and D9 (n = 3) - 1350,6?m (844 - 1688?m), and daily growth rate was 312,7?m. After the section, the 23 embryos were transfered, only one recipient mare was diagnosed as pregnant at 15 days after ovulation, however after 30 days the embryo was lost. The efficiency of the sex identification by PCR using the primers SRY and ZFX / ZFY was 85,9% (61/71), being 55,7% (34/61) determined as female: and 39,3% (27/61) as male. Embryo transfer has shown to be favorable to aid mule reproduction, whereas the recovery rate and the characteristics of the embryos are similar to equine embryos. Altough, the methodology used to section of embryos had unable the gestational development, the most part of the biopsy derived cutting allowed sexing of the embryos / A transfer?ncia de embri?es e outras biotecnologias reprodutivas s?o cada vez mais utilizadas para a produ??o de equinos e de outros animais, entretanto, mesmo com a expans?o do mercado de muares e com crescimento do n?mero de animais, as pesquisas relacionadas com a produ??o desses animais ainda s?o raras. O presente estudo avaliou as caracter?sticas relacionadas ? transfer?ncia de embri?es, a morfologia e a identifica??o do sexo de embri?es h?bridos (muares), verificando sua semelhan?a ou diverg?ncia com as caracter?sticas j? conhecidas em equinos. Foram realizadas colheitas de embri?es provenientes do cruzamento de ?guas com um jumento P?ga, nos dias 6, 7, 8 e 9 ap?s a ovula??o, a taxa de recupera??o embrion?ria, e as caracter?sticas relacionadas com a idade, morfologia e di?metro embrion?rio foram avaliadas para os diferentes dias. Ap?s essas avalia??es, uma parte dos embri?es coletados foi seccionada, com uma t?cnica adaptada onde o corte foi realizado com o aux?lio de uma l?mina de bisturi oftalmol?gico. A maior por??o resultante do corte foi destinada para transfer?ncia de embri?es e a menor parte, juntamente com os embri?es inteiros, foram utilizados para verificar a efici?ncia dos primers SRY e ZFX/ZFY, na an?lise molecular para a determina??o do sexo dos embri?es. Os dados foram analisados pelo teste Exato de Fisher, com 5% de signific?ncia, exceto o crescimento di?rio dos embri?es que foi analisado atrav?s da regress?o linear. A taxa de recupera??o embrion?ria total foi de 55,9% (71/127), e para os diferentes dias de colheita D6, D7, D8 e D9, foi 57,1% (28/49), 51% (25/49), 63% (15/24), 60% (3/5), respectivamente. Os embri?es coletados apresentavam-se nos seguintes est?gios de desenvolvimento, m?rulas: 18,3% (13/71); blastocistos iniciais: 26,8% (19/71); blastocistos: 29,6% (21/71); e blastocistos expandidos: 25,3% (18/71). O di?metro mensurado revelou que o tamanho dos embri?es variou entre 147 - 1688?m, a m?dia do di?metro de todos os embri?es recolhidos foi de 438,04?m, e de acordo com os diferentes dias de colheita, o tamanho m?dio (maior e menor di?metro) dos embri?es foi de: D6 (n = 29) ? 183,4?m (147 - 253?m), D7 (n = 24) ? 463,2?m (168 - 886?m), D8 (n = 15) ? 727,2?m (422 - 1224?m) e D9 (n = 3) ? 1350,6?m (844 ? 1688?m), e a taxa de crescimento di?ria foi de 312,7?m. Ap?s a sec??o e transfer?ncia de 23 embri?es, apenas uma receptora foi diagnosticada como gestante aos 15 dias, mas aos 30 dias, o embri?o tinha sido absorvido. A identifica??o do sexo atrav?s da t?cnica de PCR, utilizando os primers SRY e ZFx/ZFy foi de 85,9% (61/71), sendo determinado f?meas: 55,7% (34/61) e machos: 39,3% (27/61). A transfer?ncia de embri?es ? favor?vel para auxiliar na produ??o de muares, a taxa de recupera??o e as caracter?sticas dos embri?es s?o semelhantes aos embri?es equinos. A metodologia de sec??o dos embri?es, inviabilizou o desenvolvimento gestacional, entretanto a biopsia oriunda do corte permitiu a sexagem da maioria dos embri?es coletados.

Identiferoai:union.ndltd.org:IBICT/oai:localhost:jspui/1299
Date06 November 2015
CreatorsSilva, Paula Cardoso de Almeida
ContributorsJacob, J?lio Cesar Ferraz, Nogueira, Denise Monnerat, Silva, Luciano Andrade, Brand?o, Felipe Zandonadi, Soares, Maria Am?lia Menck, Mello, Marco Roberto Bourg de
PublisherUniversidade Federal Rural do Rio de Janeiro, Programa de P?s-Gradua??o em Medicina Veterin?ria (Patologia e Ci?ncias Cl?nicas), UFRRJ, Brasil, Instituto de Veterin?ria
Source SetsIBICT Brazilian ETDs
LanguagePortuguese
Detected LanguageEnglish
Typeinfo:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/doctoralThesis
Formatapplication/pdf
Sourcereponame:Biblioteca Digital de Teses e Dissertações da UFRRJ, instname:Universidade Federal Rural do Rio de Janeiro, instacron:UFRRJ
Rightsinfo:eu-repo/semantics/openAccess
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