Efeito de diferentes curvas de resfriamento, tempos de equilíbrio e crioprotetores permeáveis no congelamento de espermatozóides de caprinos
Autor(a) principal: | |
---|---|
Data de Publicação: | 2006 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | LOCUS Repositório Institucional da UFV |
Texto Completo: | http://locus.ufv.br/handle/123456789/5714 |
Resumo: | The aim of this study was to evaluate the effect of different cooling rates, equilibration times and permeating cryoprotectants on motility characteristics and plasma membrane integrity of cryopreserved goat spermatozoa. Three experiments were designed. In the first one was evaluated the effect of different cooling methods on goat semen cryopreservation. A total of 20 ejaculates from Saanen (n = 2) and Alpine (n = 2) goats were frozen using a standard dry skim milk yolk diluent with 5% glycerol. Aliquots of the extended semen were then cooled using two different cooling methods (PR1; Becker or PR2; Fürst, 2002), before being frozen in liquid nitrogen. The semen was evaluated by total motility, vigor and the response to hypoosmotic solution test (HOST), live/dead stain and thermo-resistance test (TRT), before and after thaw. In this experiment the cooling method PR2 protected cell viability better them PR1 during the criopreservation process, reducing cold shock damage and improving the cryosurvival of goat spermatozoa. In the second experiment was evaluated the effect of different equilibration times on motility characteristics and plasma membrane integrity of cryopreserved goat spermatozoa. A total of 20 ejaculates from Saanen (n = 2) and Alpine (n = 2) goats were frozen using a standard dry skim milk yolk diluent with 5% glycerol. Aliquots of the extended semen were then cooled using the cooling methodology described by Fürst (2002) and remained at the temperature of 5-4 °C by a period of 15 minutes (TE1) or 75 minutes (TE2). The semen was evaluated by total motility, vigor and the response to hypoosmotic solution test (HOST), live/dead stain and thermoresistance test (TRT), before and after thaw. In this experiment the equilibration time of 75 minutes protected the cell viability better than the period of 15 minutes during the criopreservation process, improving the cryosurvival of goat spermatozoa. In the third experiment was evaluated the effect of the glycerol (G) and ethylene glycol (EG), on motility characteristics of cryopreserved goat spermatozoa by the in vitro semen analyses. A total of 20 ejaculates from Saanen (n = 2) and Alpine (n = 2) goats were frozen using a standard dry skim milk-yolk diluent with 5% glycerol. Aliquots of the extended semen were then cooled using the cooling methodology described by Fürst (2002). The semen was evaluated by total motility, vigor and the response to hypoosmotic solution test (HOST), live/dead stain and thermo-resistance test (TRT), before and after thaw. In this experiment EG promoted a better protection of the spermatic cell viability during the cryopreservation than the G. In conclusion the use of the cooling method PR2 associated with the one hour equilibration period and the ethylene glycol improved the motility characteristics and integrity of plasma membrane, increasing the number of viable goat spermatozoa after cryopreservation. |
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Rovay, Herberthttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4762371E2Costa, Eduardo Paulino dahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787237D6Carvalho, Giovanni Ribeiro dehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4723068Z6Torres, Ciro Alexandre Alveshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787213D4Guimarães, José Domingoshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782270U6Fonseca, Jeferson Ferreira dahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4703690H82015-03-26T13:55:07Z2007-02-082015-03-26T13:55:07Z2006-08-14ROVAY, Herbert. Effect of different cooling rates, equilibration times and permeating cryoprotectants on deep-freezing of buck spermatozoa. 2006. 73 f. Dissertação (Mestrado em Genética e Melhoramento de Animais Domésticos; Nutrição e Alimentação Animal; Pastagens e Forragicul) - Universidade Federal de Viçosa, Viçosa, 2006.http://locus.ufv.br/handle/123456789/5714The aim of this study was to evaluate the effect of different cooling rates, equilibration times and permeating cryoprotectants on motility characteristics and plasma membrane integrity of cryopreserved goat spermatozoa. Three experiments were designed. In the first one was evaluated the effect of different cooling methods on goat semen cryopreservation. A total of 20 ejaculates from Saanen (n = 2) and Alpine (n = 2) goats were frozen using a standard dry skim milk yolk diluent with 5% glycerol. Aliquots of the extended semen were then cooled using two different cooling methods (PR1; Becker or PR2; Fürst, 2002), before being frozen in liquid nitrogen. The semen was evaluated by total motility, vigor and the response to hypoosmotic solution test (HOST), live/dead stain and thermo-resistance test (TRT), before and after thaw. In this experiment the cooling method PR2 protected cell viability better them PR1 during the criopreservation process, reducing cold shock damage and improving the cryosurvival of goat spermatozoa. In the second experiment was evaluated the effect of different equilibration times on motility characteristics and plasma membrane integrity of cryopreserved goat spermatozoa. A total of 20 ejaculates from Saanen (n = 2) and Alpine (n = 2) goats were frozen using a standard dry skim milk yolk diluent with 5% glycerol. Aliquots of the extended semen were then cooled using the cooling methodology described by Fürst (2002) and remained at the temperature of 5-4 °C by a period of 15 minutes (TE1) or 75 minutes (TE2). The semen was evaluated by total motility, vigor and the response to hypoosmotic solution test (HOST), live/dead stain and thermoresistance test (TRT), before and after thaw. In this experiment the equilibration time of 75 minutes protected the cell viability better than the period of 15 minutes during the criopreservation process, improving the cryosurvival of goat spermatozoa. In the third experiment was evaluated the effect of the glycerol (G) and ethylene glycol (EG), on motility characteristics of cryopreserved goat spermatozoa by the in vitro semen analyses. A total of 20 ejaculates from Saanen (n = 2) and Alpine (n = 2) goats were frozen using a standard dry skim milk-yolk diluent with 5% glycerol. Aliquots of the extended semen were then cooled using the cooling methodology described by Fürst (2002). The semen was evaluated by total motility, vigor and the response to hypoosmotic solution test (HOST), live/dead stain and thermo-resistance test (TRT), before and after thaw. In this experiment EG promoted a better protection of the spermatic cell viability during the cryopreservation than the G. In conclusion the use of the cooling method PR2 associated with the one hour equilibration period and the ethylene glycol improved the motility characteristics and integrity of plasma membrane, increasing the number of viable goat spermatozoa after cryopreservation.O objetivo deste estudo foi avaliar o efeito de diferentes protocolos de resfriamento, tempos de equilíbrio e crioprotetores permeáveis sobre as características de motilidade e integridade de membrana de espermatozóides caprinos criopreservados. Para tal, foram delineados três experimentos. No experimento 1, avaliou-se o efeito de diferentes protocolos de resfriamento sobre a criopreservação de sêmen caprino. Um total de 20 ejaculados de bodes das raças Saanen (n = 2) e Alpina (n = 2) foram congelados utilizandose um meio à base de leite desnatado-gema, acrescido de 5% de glicerol. Alíquotas do sêmen, então, diluído, foram resfriadas até 5 °C, utilizando dois protocolos de resfriamento diferentes (PR1; média de -0,12 °C/min e PR2; média de -0,4 °C/min), antes de serem congeladas em nitrogênio liquido. O sêmen foi avaliado quanto à sua motilidade espermática progressiva, vigor, taxa de recuperação da motilidade, resposta ao teste hiposmótico (HOST) e coloração. Nesse experimento, a curva de resfriamento PR2 promoveu uma melhor proteção aos efeitos causados pelo choque térmico durante a criopreservação do sêmen caprino, que a curva PR1. No experimento 2, avaliou-se o efeito de dois períodos de equilíbrio sobre as características de motilidade e integridade de membrana de espermatozóides caprinos criopreservados. Um total de 20 ejaculados de animais da raça Saanen (n = 2) e Alpina (n = 2) foram congelados utilizando-se um meio à base de leite desnatado-gema, acrescido de 5% de glicerol. Alíquotas do sêmen diluído foram, então, resfriadas segundo Fürst (2002), e mantidas a temperatura de 5 °C, por um período de 15 minutos (TE1) ou 75 minutos (TE2) antes de serem congeladas em nitrogênio liquido. O sêmen foi avaliado quanto à sua motilidade espermática progressiva, vigor, resposta ao teste hiposmótico (HOST), ao teste supravital e ao teste de termorresistência (TTR), antes e após o congelamento. Nesse experimento, o tempo de equilíbrio de 75 minutos promoveu uma melhor criopreservação do sêmen caprino que o tempo de equilíbrio curto, de 15 minutos. No experimento 3, avaliou-se o efeito do glicerol e etileno glicol sobre a criopreservação de sêmen caprino, por meio de análises in vitro do sêmen. Um total de 20 ejaculados de animais da raça Saanen (n = 2) e Alpina (n = 2) foram congelados utilizando-se um meio à base de leite desnatado-gema, acrescido de 5% de glicerol. Alíquotas do sêmen diluído foram, então, resfriadas, utilizando-se a metodologia descrita por Fürst (2002). O sêmen foi avaliado quanto à sua motilidade espermática progressiva, vigor, resposta ao teste hiposmótico (HOST), ao teste supravital e ao teste de termorresistência (TTR), antes e após o congelamento. Nesse experimento, o uso do EG como crioprotetor permeável promoveu uma melhor proteção à viabilidade da célula espermática, durante o processo de criopreservação, que o glicerol. Em conclusão, o uso do protocolo de resfriamento 2, associado a um período de equilíbrio de 1 hora e ao uso do etileno glycol, promoveu uma melhor proteção das características de motilidade e integridade de membrana, aumentando a viabilidade dos espermatozóides caprinos congelados.Conselho Nacional de Desenvolvimento Científico e Tecnológicoapplication/pdfporUniversidade Federal de ViçosaMestrado em ZootecniaUFVBRGenética e Melhoramento de Animais Domésticos; Nutrição e Alimentação Animal; Pastagens e ForragiculEquilíbrioEspermatozóideCrioprotetoresResfriamentoEquilibrationSpermatozoaCryoprotectantsCoolingCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::REPRODUCAO ANIMALEfeito de diferentes curvas de resfriamento, tempos de equilíbrio e crioprotetores permeáveis no congelamento de espermatozóides de caprinosEffect of different cooling rates, equilibration times and permeating cryoprotectants on deep-freezing of buck spermatozoainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALtexto completo.pdfapplication/pdf256712https://locus.ufv.br//bitstream/123456789/5714/1/texto%20completo.pdf96078d7a918b47a581d54f5c6c68b221MD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain127560https://locus.ufv.br//bitstream/123456789/5714/2/texto%20completo.pdf.txt3ec6040726bba2e67ac9246c8d9a4c5bMD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3763https://locus.ufv.br//bitstream/123456789/5714/3/texto%20completo.pdf.jpg36a6e68af066325b5cd01afa8da6a437MD53123456789/57142016-04-10 23:12:59.021oai:locus.ufv.br:123456789/5714Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-11T02:12:59LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false |
dc.title.por.fl_str_mv |
Efeito de diferentes curvas de resfriamento, tempos de equilíbrio e crioprotetores permeáveis no congelamento de espermatozóides de caprinos |
dc.title.alternative.eng.fl_str_mv |
Effect of different cooling rates, equilibration times and permeating cryoprotectants on deep-freezing of buck spermatozoa |
title |
Efeito de diferentes curvas de resfriamento, tempos de equilíbrio e crioprotetores permeáveis no congelamento de espermatozóides de caprinos |
spellingShingle |
Efeito de diferentes curvas de resfriamento, tempos de equilíbrio e crioprotetores permeáveis no congelamento de espermatozóides de caprinos Rovay, Herbert Equilíbrio Espermatozóide Crioprotetores Resfriamento Equilibration Spermatozoa Cryoprotectants Cooling CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::REPRODUCAO ANIMAL |
title_short |
Efeito de diferentes curvas de resfriamento, tempos de equilíbrio e crioprotetores permeáveis no congelamento de espermatozóides de caprinos |
title_full |
Efeito de diferentes curvas de resfriamento, tempos de equilíbrio e crioprotetores permeáveis no congelamento de espermatozóides de caprinos |
title_fullStr |
Efeito de diferentes curvas de resfriamento, tempos de equilíbrio e crioprotetores permeáveis no congelamento de espermatozóides de caprinos |
title_full_unstemmed |
Efeito de diferentes curvas de resfriamento, tempos de equilíbrio e crioprotetores permeáveis no congelamento de espermatozóides de caprinos |
title_sort |
Efeito de diferentes curvas de resfriamento, tempos de equilíbrio e crioprotetores permeáveis no congelamento de espermatozóides de caprinos |
author |
Rovay, Herbert |
author_facet |
Rovay, Herbert |
author_role |
author |
dc.contributor.authorLattes.por.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4762371E2 |
dc.contributor.author.fl_str_mv |
Rovay, Herbert |
dc.contributor.advisor-co1.fl_str_mv |
Costa, Eduardo Paulino da |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787237D6 |
dc.contributor.advisor-co2.fl_str_mv |
Carvalho, Giovanni Ribeiro de |
dc.contributor.advisor-co2Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4723068Z6 |
dc.contributor.advisor1.fl_str_mv |
Torres, Ciro Alexandre Alves |
dc.contributor.advisor1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787213D4 |
dc.contributor.referee1.fl_str_mv |
Guimarães, José Domingos |
dc.contributor.referee1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782270U6 |
dc.contributor.referee2.fl_str_mv |
Fonseca, Jeferson Ferreira da |
dc.contributor.referee2Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4703690H8 |
contributor_str_mv |
Costa, Eduardo Paulino da Carvalho, Giovanni Ribeiro de Torres, Ciro Alexandre Alves Guimarães, José Domingos Fonseca, Jeferson Ferreira da |
dc.subject.por.fl_str_mv |
Equilíbrio Espermatozóide Crioprotetores Resfriamento |
topic |
Equilíbrio Espermatozóide Crioprotetores Resfriamento Equilibration Spermatozoa Cryoprotectants Cooling CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::REPRODUCAO ANIMAL |
dc.subject.eng.fl_str_mv |
Equilibration Spermatozoa Cryoprotectants Cooling |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::REPRODUCAO ANIMAL |
description |
The aim of this study was to evaluate the effect of different cooling rates, equilibration times and permeating cryoprotectants on motility characteristics and plasma membrane integrity of cryopreserved goat spermatozoa. Three experiments were designed. In the first one was evaluated the effect of different cooling methods on goat semen cryopreservation. A total of 20 ejaculates from Saanen (n = 2) and Alpine (n = 2) goats were frozen using a standard dry skim milk yolk diluent with 5% glycerol. Aliquots of the extended semen were then cooled using two different cooling methods (PR1; Becker or PR2; Fürst, 2002), before being frozen in liquid nitrogen. The semen was evaluated by total motility, vigor and the response to hypoosmotic solution test (HOST), live/dead stain and thermo-resistance test (TRT), before and after thaw. In this experiment the cooling method PR2 protected cell viability better them PR1 during the criopreservation process, reducing cold shock damage and improving the cryosurvival of goat spermatozoa. In the second experiment was evaluated the effect of different equilibration times on motility characteristics and plasma membrane integrity of cryopreserved goat spermatozoa. A total of 20 ejaculates from Saanen (n = 2) and Alpine (n = 2) goats were frozen using a standard dry skim milk yolk diluent with 5% glycerol. Aliquots of the extended semen were then cooled using the cooling methodology described by Fürst (2002) and remained at the temperature of 5-4 °C by a period of 15 minutes (TE1) or 75 minutes (TE2). The semen was evaluated by total motility, vigor and the response to hypoosmotic solution test (HOST), live/dead stain and thermoresistance test (TRT), before and after thaw. In this experiment the equilibration time of 75 minutes protected the cell viability better than the period of 15 minutes during the criopreservation process, improving the cryosurvival of goat spermatozoa. In the third experiment was evaluated the effect of the glycerol (G) and ethylene glycol (EG), on motility characteristics of cryopreserved goat spermatozoa by the in vitro semen analyses. A total of 20 ejaculates from Saanen (n = 2) and Alpine (n = 2) goats were frozen using a standard dry skim milk-yolk diluent with 5% glycerol. Aliquots of the extended semen were then cooled using the cooling methodology described by Fürst (2002). The semen was evaluated by total motility, vigor and the response to hypoosmotic solution test (HOST), live/dead stain and thermo-resistance test (TRT), before and after thaw. In this experiment EG promoted a better protection of the spermatic cell viability during the cryopreservation than the G. In conclusion the use of the cooling method PR2 associated with the one hour equilibration period and the ethylene glycol improved the motility characteristics and integrity of plasma membrane, increasing the number of viable goat spermatozoa after cryopreservation. |
publishDate |
2006 |
dc.date.issued.fl_str_mv |
2006-08-14 |
dc.date.available.fl_str_mv |
2007-02-08 2015-03-26T13:55:07Z |
dc.date.accessioned.fl_str_mv |
2015-03-26T13:55:07Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
ROVAY, Herbert. Effect of different cooling rates, equilibration times and permeating cryoprotectants on deep-freezing of buck spermatozoa. 2006. 73 f. Dissertação (Mestrado em Genética e Melhoramento de Animais Domésticos; Nutrição e Alimentação Animal; Pastagens e Forragicul) - Universidade Federal de Viçosa, Viçosa, 2006. |
dc.identifier.uri.fl_str_mv |
http://locus.ufv.br/handle/123456789/5714 |
identifier_str_mv |
ROVAY, Herbert. Effect of different cooling rates, equilibration times and permeating cryoprotectants on deep-freezing of buck spermatozoa. 2006. 73 f. Dissertação (Mestrado em Genética e Melhoramento de Animais Domésticos; Nutrição e Alimentação Animal; Pastagens e Forragicul) - Universidade Federal de Viçosa, Viçosa, 2006. |
url |
http://locus.ufv.br/handle/123456789/5714 |
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por |
language |
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info:eu-repo/semantics/openAccess |
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openAccess |
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Universidade Federal de Viçosa |
dc.publisher.program.fl_str_mv |
Mestrado em Zootecnia |
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UFV |
dc.publisher.country.fl_str_mv |
BR |
dc.publisher.department.fl_str_mv |
Genética e Melhoramento de Animais Domésticos; Nutrição e Alimentação Animal; Pastagens e Forragicul |
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Universidade Federal de Viçosa |
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