Plasma seminal suíno na criopreservação de sêmen ovino

Detalhes bibliográficos
Autor(a) principal: Martins, Kauê Rodriguez
Data de Publicação: 2014
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da UFPel - Guaiaca
Texto Completo: http://guaiaca.ufpel.edu.br/handle/123456789/1234
Resumo: There is a growing interest in using artificial insemination (AI) in sheep due to the potential for genetic improvement. However, cryopreservation damages spermatozoa, decreasing their fertilizing potential when frozen semen is deposited in the cervix. Sperm damages attributed to cryopreservation may be minimized by the addition of seminal plasma (SP), which contains several factors produced by the testis, epididhymis and accessory glands with potential to prevent premature capacitation. Supplementation of SP prior to freezing would be beneficial for various processes of periods of the selection and freezing process, reported before freezing would be beneficial for the processes of selection and freezing. The supplementation of extenders with SP from ram and boars is associated with increased sperm motility after incubation in vitro, as well as when used for cooling, freezing and thawing. The objectives of this study were to test the addition of 20% boar SP to the extender to freeze ram sperm and to evaluate parameters of sperm quality after thawing. Ejaculates from four rams and three boars were collected to form pooled SP samples. A fraction of each pooled sample was used for protein quantification. Six samples from four rams were collected and diluted in Tris-egg yolk - glycerol for freezing, forming three treatments: control (no SP); inclusion of 20% ram SP; and inclusion of 20% boar SP. After thawing, the samples were subjected to a thermal stress test for five hours. Sperm quality was assessed every two hours. Analyses by flow cytometry were done to evaluate the integrity of acrosome and membrane integrity. For the control, ram SP and boar SP treatments, the evaluated parameters of sperm quality were: motility (30.4 ± 2.0, 24.6 ± 2.0 and 30.0 ± 2.0, respectively); membrane integrity (37.5 ± 2.6; 40.9 ± 2.6 and 31.4 ± 2.6 respectively); mitochondrial functionality (70.0 ± 1.7; 61.8 ± 1.7 and 63.6 ± 1.7); and DNA integrity (91.2 ± 3.1; 96 5 ± 3.1 and 93.6 ± 3.1 respectively). For those parameters, no significant differences were observed across treataments (P > 0.05). However, addition of boar SP to the extenders was related to greater acrosome integrity (59, 3 ± 3.5) than that of the control (46.7 ± 3.5) (P < 0.05), although both means were similar (P > 0.05) to that observed for the treatment with ram SP (56.7 ± 3.5). Despite of the benefit on acrosome integrity related to addition of boar SP, no other positive effects were observed for post-thawing ram sperm viability.
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spelling 2014-08-20T13:32:49Z2014-05-292014-08-20T13:32:49Z2014-03-19MARTINS, Kauê Rodriguez. Swine seminal plasma for ram sperm cryopreservation. 2014. 48 f. Dissertação (Mestrado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2014.http://guaiaca.ufpel.edu.br/handle/123456789/1234There is a growing interest in using artificial insemination (AI) in sheep due to the potential for genetic improvement. However, cryopreservation damages spermatozoa, decreasing their fertilizing potential when frozen semen is deposited in the cervix. Sperm damages attributed to cryopreservation may be minimized by the addition of seminal plasma (SP), which contains several factors produced by the testis, epididhymis and accessory glands with potential to prevent premature capacitation. Supplementation of SP prior to freezing would be beneficial for various processes of periods of the selection and freezing process, reported before freezing would be beneficial for the processes of selection and freezing. The supplementation of extenders with SP from ram and boars is associated with increased sperm motility after incubation in vitro, as well as when used for cooling, freezing and thawing. The objectives of this study were to test the addition of 20% boar SP to the extender to freeze ram sperm and to evaluate parameters of sperm quality after thawing. Ejaculates from four rams and three boars were collected to form pooled SP samples. A fraction of each pooled sample was used for protein quantification. Six samples from four rams were collected and diluted in Tris-egg yolk - glycerol for freezing, forming three treatments: control (no SP); inclusion of 20% ram SP; and inclusion of 20% boar SP. After thawing, the samples were subjected to a thermal stress test for five hours. Sperm quality was assessed every two hours. Analyses by flow cytometry were done to evaluate the integrity of acrosome and membrane integrity. For the control, ram SP and boar SP treatments, the evaluated parameters of sperm quality were: motility (30.4 ± 2.0, 24.6 ± 2.0 and 30.0 ± 2.0, respectively); membrane integrity (37.5 ± 2.6; 40.9 ± 2.6 and 31.4 ± 2.6 respectively); mitochondrial functionality (70.0 ± 1.7; 61.8 ± 1.7 and 63.6 ± 1.7); and DNA integrity (91.2 ± 3.1; 96 5 ± 3.1 and 93.6 ± 3.1 respectively). For those parameters, no significant differences were observed across treataments (P > 0.05). However, addition of boar SP to the extenders was related to greater acrosome integrity (59, 3 ± 3.5) than that of the control (46.7 ± 3.5) (P < 0.05), although both means were similar (P > 0.05) to that observed for the treatment with ram SP (56.7 ± 3.5). Despite of the benefit on acrosome integrity related to addition of boar SP, no other positive effects were observed for post-thawing ram sperm viability.O interesse pelo uso da inseminação artificial (IA) em ovinos vem crescendo, em função do avanço no melhoramento genético. Entretanto, a criopreservação causa danos aos espermatozóides, diminuindo seu potencial fertilizante, quando a IA é feita com sêmen congelado, pela via cervical. Uma alternativa para proteger ou recuperar a célula dos danos da criopreservação é a adição de plasma seminal (PS), que contém vários fatores produzidos pelos testículos, epidídimos e glândulas acessórias do macho, com potencial de prevenir a capacitação prematura e danos gerados pelo congelamento. A adição de PS antes do congelamento seria benéfica para os processos de seleção e congelamento. A suplementação do diluente com PS ovino e suíno foi associada com aumento na motilidade espermática, após a segunda hora de incubação sob condições in vitro, assim como quando usado na refrigeração, congelamento e descongelamento. Este estudo teve como objetivo testar a adição de PS suíno (20%) ao diluente para congelamento do sêmen ovino e avaliar os parâmetros seminais in vitro pós-descongelamento. Ejaculados de quatro machos ovinos e três machos suínos foram coletados para formação de amostras combinando PS de vários machos (pools). Uma fração das amostras de PS foi destinada a quantificação de proteínas. Seis coletas dos quatro machos ovinos foram colhidas e diluídas em Tris-gema de ovo-glicerol, para congelamento, compondo três tratamentos: controle (sem PS); inclusão de 20% de PS ovino; e inclusão de 20% PS suíno. Após o descongelamento, as amostras foram submetidas a um teste de termo resistência durante cinco horas. Avaliações de qualidade espermática foram realizadas a cada duas horas. Também foram realizadas análises por citometria de fluxo para as avaliações de integridade de acrossoma e integridade de membrana. Para os tratamentos controle, PS ovino e PS suíno, não foram observadas diferenças (P > 0.05) quanto a motilidade (30,0 ± 2,0; 30,4 ± 2,0 e 24,6 ± 2,0 respectivamente), integridade de membrana (37,5 ± 2,6; 40,9 ± 2,6 e 31,4 ± 2,6, respectivamente), função mitocondrial (70,0 ± 1,7; 61,8 ± 1,7 e 63,6 ± 1,7, respectivamente) e integridade de DNA (91,2 ± 3,1; 96,5 ± 3,1 e 93,6 ± 3,1, respectivamente). A integridade do acrossoma foi maior (P < 0.05) com inclusão de PS suíno (59,3 ± 3,5) em comparação com o controle (46,7 ± 3,5), mas ambas as médias foram similares (P > 0.05) à observada para o PS ovino (56,7 ± 3,5). Conclui-se que apesar do beneficio do PS suíno a 20% para a integridade de acrossoma, não se obteve resultados positivos nas demais avaliações de qualidade seminal.application/pdfporUniversidade Federal de PelotasPrograma de Pós-Graduação em BiotecnologiaUFPelBRBiotecnologiaFreezingSeminal plasmaBoarRamFlow-cytometryDecapacitationCongelamentoPlasma seminalSuínoOvinoCitometriaDecapitaçãoCNPQ::CIENCIAS BIOLOGICAS::GENETICA::GENETICA ANIMALPlasma seminal suíno na criopreservação de sêmen ovinoSwine seminal plasma for ram sperm cryopreservationinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesishttp://lattes.cnpq.br/7793098173274177http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4793234U8Lucia Júnior, ThomazMartins, Kauê Rodriguezinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFPel - Guaiacainstname:Universidade Federal de Pelotas (UFPEL)instacron:UFPELORIGINALdissertacao_kaue_rodriguez_martins.pdfapplication/pdf452926http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1234/1/dissertacao_kaue_rodriguez_martins.pdfae46017afbe7b23ca7e1d34a015987faMD51open accessTEXTdissertacao_kaue_rodriguez_martins.pdf.txtdissertacao_kaue_rodriguez_martins.pdf.txtExtracted Texttext/plain76601http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1234/2/dissertacao_kaue_rodriguez_martins.pdf.txta6be24c1aa40b7683af01be8da9cdd52MD52open accessTHUMBNAILdissertacao_kaue_rodriguez_martins.pdf.jpgdissertacao_kaue_rodriguez_martins.pdf.jpgGenerated Thumbnailimage/jpeg1780http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1234/3/dissertacao_kaue_rodriguez_martins.pdf.jpg1767318a887a668d0d8de771f1fe7741MD53open access123456789/12342019-08-23 10:31:13.648open accessoai:guaiaca.ufpel.edu.br:123456789/1234Repositório InstitucionalPUBhttp://repositorio.ufpel.edu.br/oai/requestrippel@ufpel.edu.br || repositorio@ufpel.edu.br || aline.batista@ufpel.edu.bropendoar:2019-08-23T13:31:13Repositório Institucional da UFPel - Guaiaca - Universidade Federal de Pelotas (UFPEL)false
dc.title.por.fl_str_mv Plasma seminal suíno na criopreservação de sêmen ovino
dc.title.alternative.eng.fl_str_mv Swine seminal plasma for ram sperm cryopreservation
title Plasma seminal suíno na criopreservação de sêmen ovino
spellingShingle Plasma seminal suíno na criopreservação de sêmen ovino
Martins, Kauê Rodriguez
Freezing
Seminal plasma
Boar
Ram
Flow-cytometry
Decapacitation
Congelamento
Plasma seminal
Suíno
Ovino
Citometria
Decapitação
CNPQ::CIENCIAS BIOLOGICAS::GENETICA::GENETICA ANIMAL
title_short Plasma seminal suíno na criopreservação de sêmen ovino
title_full Plasma seminal suíno na criopreservação de sêmen ovino
title_fullStr Plasma seminal suíno na criopreservação de sêmen ovino
title_full_unstemmed Plasma seminal suíno na criopreservação de sêmen ovino
title_sort Plasma seminal suíno na criopreservação de sêmen ovino
author Martins, Kauê Rodriguez
author_facet Martins, Kauê Rodriguez
author_role author
dc.contributor.authorLattes.por.fl_str_mv http://lattes.cnpq.br/7793098173274177
dc.contributor.advisorLattes.por.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4793234U8
dc.contributor.advisor1.fl_str_mv Lucia Júnior, Thomaz
dc.contributor.author.fl_str_mv Martins, Kauê Rodriguez
contributor_str_mv Lucia Júnior, Thomaz
dc.subject.eng.fl_str_mv Freezing
Seminal plasma
Boar
Ram
Flow-cytometry
Decapacitation
topic Freezing
Seminal plasma
Boar
Ram
Flow-cytometry
Decapacitation
Congelamento
Plasma seminal
Suíno
Ovino
Citometria
Decapitação
CNPQ::CIENCIAS BIOLOGICAS::GENETICA::GENETICA ANIMAL
dc.subject.por.fl_str_mv Congelamento
Plasma seminal
Suíno
Ovino
Citometria
Decapitação
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS BIOLOGICAS::GENETICA::GENETICA ANIMAL
description There is a growing interest in using artificial insemination (AI) in sheep due to the potential for genetic improvement. However, cryopreservation damages spermatozoa, decreasing their fertilizing potential when frozen semen is deposited in the cervix. Sperm damages attributed to cryopreservation may be minimized by the addition of seminal plasma (SP), which contains several factors produced by the testis, epididhymis and accessory glands with potential to prevent premature capacitation. Supplementation of SP prior to freezing would be beneficial for various processes of periods of the selection and freezing process, reported before freezing would be beneficial for the processes of selection and freezing. The supplementation of extenders with SP from ram and boars is associated with increased sperm motility after incubation in vitro, as well as when used for cooling, freezing and thawing. The objectives of this study were to test the addition of 20% boar SP to the extender to freeze ram sperm and to evaluate parameters of sperm quality after thawing. Ejaculates from four rams and three boars were collected to form pooled SP samples. A fraction of each pooled sample was used for protein quantification. Six samples from four rams were collected and diluted in Tris-egg yolk - glycerol for freezing, forming three treatments: control (no SP); inclusion of 20% ram SP; and inclusion of 20% boar SP. After thawing, the samples were subjected to a thermal stress test for five hours. Sperm quality was assessed every two hours. Analyses by flow cytometry were done to evaluate the integrity of acrosome and membrane integrity. For the control, ram SP and boar SP treatments, the evaluated parameters of sperm quality were: motility (30.4 ± 2.0, 24.6 ± 2.0 and 30.0 ± 2.0, respectively); membrane integrity (37.5 ± 2.6; 40.9 ± 2.6 and 31.4 ± 2.6 respectively); mitochondrial functionality (70.0 ± 1.7; 61.8 ± 1.7 and 63.6 ± 1.7); and DNA integrity (91.2 ± 3.1; 96 5 ± 3.1 and 93.6 ± 3.1 respectively). For those parameters, no significant differences were observed across treataments (P > 0.05). However, addition of boar SP to the extenders was related to greater acrosome integrity (59, 3 ± 3.5) than that of the control (46.7 ± 3.5) (P < 0.05), although both means were similar (P > 0.05) to that observed for the treatment with ram SP (56.7 ± 3.5). Despite of the benefit on acrosome integrity related to addition of boar SP, no other positive effects were observed for post-thawing ram sperm viability.
publishDate 2014
dc.date.accessioned.fl_str_mv 2014-08-20T13:32:49Z
dc.date.available.fl_str_mv 2014-05-29
2014-08-20T13:32:49Z
dc.date.issued.fl_str_mv 2014-03-19
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
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dc.identifier.citation.fl_str_mv MARTINS, Kauê Rodriguez. Swine seminal plasma for ram sperm cryopreservation. 2014. 48 f. Dissertação (Mestrado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2014.
dc.identifier.uri.fl_str_mv http://guaiaca.ufpel.edu.br/handle/123456789/1234
identifier_str_mv MARTINS, Kauê Rodriguez. Swine seminal plasma for ram sperm cryopreservation. 2014. 48 f. Dissertação (Mestrado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2014.
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