Avaliação individual e racial da qualidade do sêmen canino in natura e criopreservado com diferentes protocolos
Autor(a) principal: | |
---|---|
Data de Publicação: | 2008 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | LOCUS Repositório Institucional da UFV |
Texto Completo: | http://locus.ufv.br/handle/123456789/4975 |
Resumo: | Although the first birth of pups with use of frozen semen has been reported almost 40 years ago, still today the conception rate with thawed canine semen is generally inferior to the observed in other species. Many factors influencing the semen quality through the cryopreservation process are known, including the centrifugation protocol, the extender composition, the association between the cryoprotectors, the cooling rate, the time of equilibrium and the freezing rate, as well as the association between thawing and freezing rates. Individual variations in the semen freezability are found in many species, including the dog. Some authors attribute these variations to the differences in the spermatic membrane lipid composition and spermatozoon sensitivity of the glycerol toxic effects. In present work 36 semen samples where collected, from 12 dog of the: Beagle, Schnauzer, Doberman and Boxer breeds, by digital manipulation. After the collection the semen was centrifuged, reextended in Tris-Citrato extender added with 1, 2, 4 and 6% of glycerol and packaged in 0,25mL straws. After that the semen were cooled for 1 hour until 4oC, then half of the straws of each treatment was immediately freezed and the remain was kept by more 1 hour in equilibrium at 4oC before the freezing. Two freezing rates had been tested in termical box contend liquid nitrogen through the positioning of the straws 5 and 10 cm above the surface of nitrogen per 10 minutes. The thawing was carried through immersion in water 38oC per 1 minute. The semen was evaluated at the moment of the collection, after the centrifugation and immediately after thawing for spermatozoa movement through motility, progressive status, and sperm index and for plasma membrane integrity and viability through hiposmotic swelling tests and live/dead stain. The sperm longevity was evaluated through the thermorresistece test. In the present work the individual averages of sperm motility and progressive status in the fresh semen had varied from 3.5 to 5.0 and 63 to 90% respectively, while the breed averages had varied of 4.4 the 4.9 and 80 to 90% respectively. The semen centrifugation resulted in significant fall of 13% in progressive status and 14% in the motility and spermatic index. The analysis of the parameters of plasma membrane viability and integrity in the semen after centrifugation shows significant reduction of 38% and 14% in the reactive cells to the hiposmotic swelling test and to the supravital coloration respectively. The results of the hiposmotic swelling test, live/dead stain and thermorresistence in the thawed semen demonstrate better performance when protocols of freezing were used with equilibrium time. Influence of the freezing rate, in isolated form, was not observed on the seminal parameters evaluated and significant variations had been only observed when equilibrium time and glycerol concentration were associated. Isolate the increase in the percentage of glycerol from 1 to 6% in the extender composition, promotes prompt improvement to plasma membrane integrity and to spermatic index in the thawed samples. However, when associates the observed results of biggest levels of glycerol to the presence of equilibrium time and to the slow rate of freezing, significant increases are observed in all the studied parameters. Individual variation was observed in spermatic index of thawed semen, but not in the fresh semen. Already the parameters of plasma membrane integrity and viability had presented individual variation in both. When the parameter breeds is evaluated, any significant alterations were observed on the evaluated parameters in fresh semen, however in the thawed semen, significant variations in the plasma membrane integrity and viability had been observed. The dogs of the Schnauzer breed had presented significantly shorter sperm longevity after the thawing. The Doberman and Boxer breeds had presented the best results of freezability in in vitro evaluation. With the exception of the test of live/dead stain, the studied parameters had presented significant correlations between the data collected in fresh semen with those observed in thawed semen. |
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Mascarenhas, Rebeca Marqueshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4776958J3Costa, Eduardo Paulino dahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787237D6Matta, Sérgio Luis Pinto dahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4798314Z0Paula, Tarcízio Antônio Rego dehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4701637D5Costa, Deiler Sampaiohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4790473Y9Cunha, Isabel Candia Nunes dahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4773817A7Valle, Guilherme Ribeirohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4790271Z4Guimarães, José Domingoshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782270U62015-03-26T13:46:40Z2009-08-122015-03-26T13:46:40Z2008-02-08MASCARENHAS, Rebeca Marques. Individual and breed evaluation of the quality of canine sêmen in natura and cryopreserved with different protocols. 2008. 83 f. Dissertação (Mestrado em Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de) - Universidade Federal de Viçosa, Viçosa, 2008.http://locus.ufv.br/handle/123456789/4975Although the first birth of pups with use of frozen semen has been reported almost 40 years ago, still today the conception rate with thawed canine semen is generally inferior to the observed in other species. Many factors influencing the semen quality through the cryopreservation process are known, including the centrifugation protocol, the extender composition, the association between the cryoprotectors, the cooling rate, the time of equilibrium and the freezing rate, as well as the association between thawing and freezing rates. Individual variations in the semen freezability are found in many species, including the dog. Some authors attribute these variations to the differences in the spermatic membrane lipid composition and spermatozoon sensitivity of the glycerol toxic effects. In present work 36 semen samples where collected, from 12 dog of the: Beagle, Schnauzer, Doberman and Boxer breeds, by digital manipulation. After the collection the semen was centrifuged, reextended in Tris-Citrato extender added with 1, 2, 4 and 6% of glycerol and packaged in 0,25mL straws. After that the semen were cooled for 1 hour until 4oC, then half of the straws of each treatment was immediately freezed and the remain was kept by more 1 hour in equilibrium at 4oC before the freezing. Two freezing rates had been tested in termical box contend liquid nitrogen through the positioning of the straws 5 and 10 cm above the surface of nitrogen per 10 minutes. The thawing was carried through immersion in water 38oC per 1 minute. The semen was evaluated at the moment of the collection, after the centrifugation and immediately after thawing for spermatozoa movement through motility, progressive status, and sperm index and for plasma membrane integrity and viability through hiposmotic swelling tests and live/dead stain. The sperm longevity was evaluated through the thermorresistece test. In the present work the individual averages of sperm motility and progressive status in the fresh semen had varied from 3.5 to 5.0 and 63 to 90% respectively, while the breed averages had varied of 4.4 the 4.9 and 80 to 90% respectively. The semen centrifugation resulted in significant fall of 13% in progressive status and 14% in the motility and spermatic index. The analysis of the parameters of plasma membrane viability and integrity in the semen after centrifugation shows significant reduction of 38% and 14% in the reactive cells to the hiposmotic swelling test and to the supravital coloration respectively. The results of the hiposmotic swelling test, live/dead stain and thermorresistence in the thawed semen demonstrate better performance when protocols of freezing were used with equilibrium time. Influence of the freezing rate, in isolated form, was not observed on the seminal parameters evaluated and significant variations had been only observed when equilibrium time and glycerol concentration were associated. Isolate the increase in the percentage of glycerol from 1 to 6% in the extender composition, promotes prompt improvement to plasma membrane integrity and to spermatic index in the thawed samples. However, when associates the observed results of biggest levels of glycerol to the presence of equilibrium time and to the slow rate of freezing, significant increases are observed in all the studied parameters. Individual variation was observed in spermatic index of thawed semen, but not in the fresh semen. Already the parameters of plasma membrane integrity and viability had presented individual variation in both. When the parameter breeds is evaluated, any significant alterations were observed on the evaluated parameters in fresh semen, however in the thawed semen, significant variations in the plasma membrane integrity and viability had been observed. The dogs of the Schnauzer breed had presented significantly shorter sperm longevity after the thawing. The Doberman and Boxer breeds had presented the best results of freezability in in vitro evaluation. With the exception of the test of live/dead stain, the studied parameters had presented significant correlations between the data collected in fresh semen with those observed in thawed semen.O presente trabalho busca descrever a análise in vitro do sêmen fresco e as diferenças individuais e raciais na congelabilidade do sêmen de cães das raças Beagle, Schnauzer, Doberman e Boxer. Foi também avaliado o efeito da centrifugação, do tempo de equilíbrio, da taxa de congelamento e de diferentes concentrações de glicerol, sobre a qualidade o sêmen canino criopreservado em meio diluidor a base de Tris-Citrato. Foram ainda analisadas as correlação dos parâmetros de avaliação in vitro do sêmen antes e depois o congelamento. Para tanto, foram utilizados 12 cães das raças Beagle (n=3), Schnauzer (n=3), Doberman (n=3) e Boxer (n=3), sendo 3 ejaculados coletados por indivíduo, totalizando 36 ejaculados. Após a coleta o sêmen foi centrifugado, ressuspendido em meio Tris-Citrato contendo 1, 2, 4 e 6% de glicerol e evasado em palhetas de 0,25mL. Após o envase o sêmen foi resfriado por 1 hora a 40C e em seguida metade das palhetas de cada tratamento foi encaminhada para o congelamento e o restante foi mantido por mais 1 hora em equilíbrio a 4oC, antes do congelamento. Duas taxas de congelamento foram testadas em caixa de isopor contendo nitrogênio líquido através do posicionamento das palhetas a 5 e 10 cm da superfície do nitrogênio por 10 minutos. O descongelamento foi realizado através de imersão em água a 38oC por 1 minuto. O sêmen foi avaliado no momento da coleta, após a centrifugação e ao descongelamento quanto à movimentação dos espermatozóides através de vigor, motilidade e índice espermático e quanto à integridade e viabilidade da membrana plasmática pelos testes hiposmótico e de coloração supravital. A longevidade espermática foi estimada no sêmen descongelado através do teste de termorresistência (TTR). No presente trabalho as médias individuais de vigor e motilidade espermáticos no sêmen fresco variaram de 3,5 a 5,0 e de 63 a 90% respectivamente enquanto as médias raciais variaram de 4,4 a 4,9 e de 80 a 90% respectivamente. A centrifugação do sêmen resultou em queda de cerca de 13% no vigor espermático, 14% na motilidade e no índice espermático. A análise dos parâmetros de viabilidade e integridade de membrana plasmática no sêmen após centrifugação mostra diminuição de 38% e 14% nas células reativas ao teste hiposmótico e de coloração supravital respectivamente. Os resultados dos testes hiposmótico, de coloração supravital e de termorresistência no sêmen descongelado demonstram de maneira geral melhor desempenho dos protocolos de congelamento com a presença do tempo de equilíbrio. Não foi observada influência da taxa de congelamento de forma isolada sobre os parâmetros seminais avaliados. Variações significativas só foram observadas quando se associou à variação da taxa de congelamento, variações no tempo de equilíbrio e concentração de glicerol. Isoladamente, o aumento no percentual de glicerol de 1 para 6% na composição do meio diluidor, promove melhoria pontual na integridade da membrana citoplasmática e no índice espermático ao descongelamento. Porém, quando se associa os resultados observados dos maiores níveis de glicerol à presença do tempo de equilíbrio e à taxa lenta de congelamento, aumentos significativos são observados em todos os parâmetros estudados. Variação individual foi observada no índice espermático no sêmen descongelado, mas não no sêmen fresco. Já os parâmetros de integridade e viabilidade da membrana plasmática apresentaram variação individual em ambos. Quando se avalia a variável raça não se observa alterações significativas quanto aos parâmetros avaliados no sêmen fresco, porém no sêmen descongelado, observaram-se variações significativas na integridade e viabilidade da membrana plasmática. Os cães da raça Schnauzer apresentaram significativamente menor longevidade espermática após o descongelamento. As raças Doberman e Boxer apresentaram os melhores resultados de congelabilidade na avaliação in vitro. A exceção do teste de coloração supravital, os parâmetros estudados apresentaram correlações significativas entre os dados coletados no sêmen fresco com aqueles observados no sêmen descongelado.Coordenação de Aperfeiçoamento de Pessoal de Nível Superiorapplication/pdfporUniversidade Federal de ViçosaMestrado em Medicina VeterináriaUFVBRBiotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. deSêmenCriopreservaçãoCãoSêmenCryopreservationDogsCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::REPRODUCAO ANIMALAvaliação individual e racial da qualidade do sêmen canino in natura e criopreservado com diferentes protocolosIndividual and breed evaluation of the quality of canine sêmen in natura and cryopreserved with different protocolsinfo: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/pdf702747https://locus.ufv.br//bitstream/123456789/4975/1/texto%20completo.pdf881b24ba4fe9f249940d1ad2d3923f73MD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain165847https://locus.ufv.br//bitstream/123456789/4975/2/texto%20completo.pdf.txtf266656883b085504b697e7c51559828MD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3529https://locus.ufv.br//bitstream/123456789/4975/3/texto%20completo.pdf.jpg6203dde34c6771fb1650ce832a71cf4eMD53123456789/49752016-04-11 23:05:28.189oai:locus.ufv.br:123456789/4975Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-12T02:05:28LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false |
dc.title.por.fl_str_mv |
Avaliação individual e racial da qualidade do sêmen canino in natura e criopreservado com diferentes protocolos |
dc.title.alternative.eng.fl_str_mv |
Individual and breed evaluation of the quality of canine sêmen in natura and cryopreserved with different protocols |
title |
Avaliação individual e racial da qualidade do sêmen canino in natura e criopreservado com diferentes protocolos |
spellingShingle |
Avaliação individual e racial da qualidade do sêmen canino in natura e criopreservado com diferentes protocolos Mascarenhas, Rebeca Marques Sêmen Criopreservação Cão Sêmen Cryopreservation Dogs CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::REPRODUCAO ANIMAL |
title_short |
Avaliação individual e racial da qualidade do sêmen canino in natura e criopreservado com diferentes protocolos |
title_full |
Avaliação individual e racial da qualidade do sêmen canino in natura e criopreservado com diferentes protocolos |
title_fullStr |
Avaliação individual e racial da qualidade do sêmen canino in natura e criopreservado com diferentes protocolos |
title_full_unstemmed |
Avaliação individual e racial da qualidade do sêmen canino in natura e criopreservado com diferentes protocolos |
title_sort |
Avaliação individual e racial da qualidade do sêmen canino in natura e criopreservado com diferentes protocolos |
author |
Mascarenhas, Rebeca Marques |
author_facet |
Mascarenhas, Rebeca Marques |
author_role |
author |
dc.contributor.authorLattes.por.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4776958J3 |
dc.contributor.author.fl_str_mv |
Mascarenhas, Rebeca Marques |
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 |
Matta, Sérgio Luis Pinto da |
dc.contributor.advisor-co2Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4798314Z0 |
dc.contributor.advisor1.fl_str_mv |
Paula, Tarcízio Antônio Rego de |
dc.contributor.advisor1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4701637D5 |
dc.contributor.referee1.fl_str_mv |
Costa, Deiler Sampaio |
dc.contributor.referee1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4790473Y9 |
dc.contributor.referee2.fl_str_mv |
Cunha, Isabel Candia Nunes da |
dc.contributor.referee2Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4773817A7 |
dc.contributor.referee3.fl_str_mv |
Valle, Guilherme Ribeiro |
dc.contributor.referee3Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4790271Z4 |
dc.contributor.referee4.fl_str_mv |
Guimarães, José Domingos |
dc.contributor.referee4Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782270U6 |
contributor_str_mv |
Costa, Eduardo Paulino da Matta, Sérgio Luis Pinto da Paula, Tarcízio Antônio Rego de Costa, Deiler Sampaio Cunha, Isabel Candia Nunes da Valle, Guilherme Ribeiro Guimarães, José Domingos |
dc.subject.por.fl_str_mv |
Sêmen Criopreservação Cão |
topic |
Sêmen Criopreservação Cão Sêmen Cryopreservation Dogs CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::REPRODUCAO ANIMAL |
dc.subject.eng.fl_str_mv |
Sêmen Cryopreservation Dogs |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::REPRODUCAO ANIMAL |
description |
Although the first birth of pups with use of frozen semen has been reported almost 40 years ago, still today the conception rate with thawed canine semen is generally inferior to the observed in other species. Many factors influencing the semen quality through the cryopreservation process are known, including the centrifugation protocol, the extender composition, the association between the cryoprotectors, the cooling rate, the time of equilibrium and the freezing rate, as well as the association between thawing and freezing rates. Individual variations in the semen freezability are found in many species, including the dog. Some authors attribute these variations to the differences in the spermatic membrane lipid composition and spermatozoon sensitivity of the glycerol toxic effects. In present work 36 semen samples where collected, from 12 dog of the: Beagle, Schnauzer, Doberman and Boxer breeds, by digital manipulation. After the collection the semen was centrifuged, reextended in Tris-Citrato extender added with 1, 2, 4 and 6% of glycerol and packaged in 0,25mL straws. After that the semen were cooled for 1 hour until 4oC, then half of the straws of each treatment was immediately freezed and the remain was kept by more 1 hour in equilibrium at 4oC before the freezing. Two freezing rates had been tested in termical box contend liquid nitrogen through the positioning of the straws 5 and 10 cm above the surface of nitrogen per 10 minutes. The thawing was carried through immersion in water 38oC per 1 minute. The semen was evaluated at the moment of the collection, after the centrifugation and immediately after thawing for spermatozoa movement through motility, progressive status, and sperm index and for plasma membrane integrity and viability through hiposmotic swelling tests and live/dead stain. The sperm longevity was evaluated through the thermorresistece test. In the present work the individual averages of sperm motility and progressive status in the fresh semen had varied from 3.5 to 5.0 and 63 to 90% respectively, while the breed averages had varied of 4.4 the 4.9 and 80 to 90% respectively. The semen centrifugation resulted in significant fall of 13% in progressive status and 14% in the motility and spermatic index. The analysis of the parameters of plasma membrane viability and integrity in the semen after centrifugation shows significant reduction of 38% and 14% in the reactive cells to the hiposmotic swelling test and to the supravital coloration respectively. The results of the hiposmotic swelling test, live/dead stain and thermorresistence in the thawed semen demonstrate better performance when protocols of freezing were used with equilibrium time. Influence of the freezing rate, in isolated form, was not observed on the seminal parameters evaluated and significant variations had been only observed when equilibrium time and glycerol concentration were associated. Isolate the increase in the percentage of glycerol from 1 to 6% in the extender composition, promotes prompt improvement to plasma membrane integrity and to spermatic index in the thawed samples. However, when associates the observed results of biggest levels of glycerol to the presence of equilibrium time and to the slow rate of freezing, significant increases are observed in all the studied parameters. Individual variation was observed in spermatic index of thawed semen, but not in the fresh semen. Already the parameters of plasma membrane integrity and viability had presented individual variation in both. When the parameter breeds is evaluated, any significant alterations were observed on the evaluated parameters in fresh semen, however in the thawed semen, significant variations in the plasma membrane integrity and viability had been observed. The dogs of the Schnauzer breed had presented significantly shorter sperm longevity after the thawing. The Doberman and Boxer breeds had presented the best results of freezability in in vitro evaluation. With the exception of the test of live/dead stain, the studied parameters had presented significant correlations between the data collected in fresh semen with those observed in thawed semen. |
publishDate |
2008 |
dc.date.issued.fl_str_mv |
2008-02-08 |
dc.date.available.fl_str_mv |
2009-08-12 2015-03-26T13:46:40Z |
dc.date.accessioned.fl_str_mv |
2015-03-26T13:46:40Z |
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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masterThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
MASCARENHAS, Rebeca Marques. Individual and breed evaluation of the quality of canine sêmen in natura and cryopreserved with different protocols. 2008. 83 f. Dissertação (Mestrado em Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de) - Universidade Federal de Viçosa, Viçosa, 2008. |
dc.identifier.uri.fl_str_mv |
http://locus.ufv.br/handle/123456789/4975 |
identifier_str_mv |
MASCARENHAS, Rebeca Marques. Individual and breed evaluation of the quality of canine sêmen in natura and cryopreserved with different protocols. 2008. 83 f. Dissertação (Mestrado em Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de) - Universidade Federal de Viçosa, Viçosa, 2008. |
url |
http://locus.ufv.br/handle/123456789/4975 |
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por |
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Universidade Federal de Viçosa |
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Mestrado em Medicina Veterinária |
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UFV |
dc.publisher.country.fl_str_mv |
BR |
dc.publisher.department.fl_str_mv |
Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de |
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Universidade Federal de Viçosa |
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