Uso do extrato líquido de melancia (Citrullus lanatus) na criopreservação de sêmen de touros da raça Nelore

Detalhes bibliográficos
Autor(a) principal: Castilho, Erick Fonseca de
Data de Publicação: 2012
Tipo de documento: Tese
Idioma: por
Título da fonte: LOCUS Repositório Institucional da UFV
Texto Completo: http://locus.ufv.br/handle/123456789/1460
Resumo: The objectives of this study were to evaluate the potential of liquid watermelon extract as a natural diluent for cooling and/or freezing semen, and determining the cryoprotectant effect of liquid watermelon extract on the integrity of the plasma membrane of bovine sperm. Semen from four adult bulls were used, totalizing 10 semen samples for each one. After semen collection, the evaluation consisted of physical and morphological exam, live and dead cells (supravital test) and hypoosmotic swelling test. Afterwards, the fresh semen was divided in four equal parts and diluted with respective extenders: D1 (Bovimix® - control); D2 (Bovimix® base + 25 % of liquid watermelon extract); D3 (Bovimix® base + 50 % of liquid watermelon extract); and D4 (100 % of liquid watermelon extract base). After final dilutions, sperm motility and vigor were evaluated in each extender, and posterior seal. The straws were cooled and frozen in the freezing machine (Cryogen Dualflex®). Then, the straws were immersed in nitrogen. The thawing of the samples was done by immersion of the straws in a 37 °C water-bath for 30 minutes to evaluate the sperm motility and vigor, morphological characteristics of sperm, supravital test, hypoosmotic swelling test, epifluorescence test and thermoresistence test. The volume, motility, sperm vigor, sperm concentration per mL, supravital test, hypoosmotic swelling test, major and total sperm defects varied among animals, but showed no difference between them (P>0.05). The appearance, whirlpool and minor sperm defects showed differences among animals (P<0.05). Sperm motility showed high and positive correlation (r = 0.98) with the hypoosmotic swelling test. The supravital test showed high and positive correlation (r = 0.88) with the hypoosmotic swelling test. There was an average and negative correlation (r = -0.48) between motility and sperm concentration per mL, as well as average and negative correlation (r = -0.46) between the hypoosmotic swelling test and sperm concentration per mL. The motility and sperm vigor of the diluted semen (pre-cooling) in all extenders used, presented no difference between them (P>0.05). The motility upon thawing and at the end of thermoresistence test differed between extenders (P<0.05), where higher values were obtained by diluents D1 and D2, and lower values were obtained by diluents D3 and D4. At the beginning of the thermoresistence test, there was no difference between diluents D1 and D2. However, there was a difference when comparing the diluent D1 with D3 and D4. There was no difference between the diluent D2 and D3 or D2 and D4. At the end of the thermoresistence test, there was no difference between the diluents D1 and D2 or D1 and D4, but there was a difference between D1 and D3. There was no difference when comparing the diluent D2 with D3 and D4. The average sperm alive in the supravital test at the time of thawing in all extenders used, presented no difference between them (P>0.05). During the thermoresistence test, the average vigor of postthawed semen, in all extenders used, showed no difference between them (P>0.05). The hypoosmotic swelling test differed between extenders (P<0.05), where higher values were recorded in the diluent D1, an intermediate value was recorded in the diluent D2 and lower values were recorded in the diluents D3 and D4. There was no difference when comparing the diluent D1 with D2 and, no difference in between the diluents D2 and D3 or D2 and D4. However, there was a difference when comparing the diluent D1 with D3 and D4. There was no difference between the diluents D2 and D3 or D4. The epifluorescence test differed between extenders (P<0.05), but there was no difference between diluents D1 and D2. There was no difference between the diluents D2 and D3, or between the diluents D3 and D4. However, there was a difference when comparing D1 with D3 and D4. There were differences between diluents D2 and D4. The major and total sperm defects ranged among extenders, but showed no difference between them (P>0.05). However, minor sperm defects differed between extenders (P<0.05), where the highest values were observed in the diluent D3, and lower values were observed in the other diluents. There was no difference between diluent D1 and the other extenders. However, there was a difference between diluents D3 and D4. In the diluent D1, the values obtained in the supravital test showed average and positive correlation (r = 0.49) with sperm motility. The hypoosmotic swelling test showed high and positive correlation (r = 0.89) with intact sperm and, high and negative correlation (r = -0.80) with damaged spermatozoa observed in epifluorescence test. In the diluent D2, the values obtained in hypoosmotic test showed high and positive correlation (r = 0.86) with intact sperm, high and negative correlation (r = -0.83) with damaged spermatozoa and, average and positive correlation (r = 0.41) with semi-damaged sperm observed in epifluorescence test. In the diluent 3, the values obtained in the supravital test showed low and positive correlation (r = 0.33) with sperm motility. The hypoosmotic test showed high and positive correlation (r = 0.82) with sperm integrity, high and negative correlation (r = -0.80) with damaged sperm and, average and positive correlation (r = 0.51) with semi-damaged sperm observed in epifluorescence test. In the diluent D4, the values obtained in the supravital test showed average and positive correlation (r = 0.46) with sperm motility. The hypoosmotic swelling test showed high and positive correlation (r = 0.83) with intact sperm, high and negative correlation (r = - 0.85) with damaged sperm and, low and positive correlation (r = 0.38) with semidamaged sperm observed in epifluorescence test. At concentrations of 50 % and 100 %, the liquid watermelon extract was not effective in maintaining the integrity and sperm viability after thawing. At the concentration of 25 %, the liquid watermelon extract maintained the structural integrity of the membrane of spermatozoa during the cryopreservation process, and viable after the thermoresistence test, could be an alternative in the composition of diluents for cryopreservation of bovine semen.
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spelling Castilho, Erick Fonseca dehttp://lattes.cnpq.br/7525440848644647Costa, Eduardo Paulino dahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787237D6Fonseca, Jeferson Ferreira dahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4703690H8Guimarães, José Domingoshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782270U6Siqueira, Jeanne Brochhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4766315A3Paula, Tarcízio Antônio Rego dehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4701637D5Guimarães, Simone Eliza Facionihttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782526Y2Torres, Ciro Alexandre Alveshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787213D42015-03-26T12:47:49Z2013-11-292015-03-26T12:47:49Z2012-05-28CASTILHO, Erick Fonseca de. Use of liquid watermelon extract (Citrullus lanatus) in cryopreservation of semen of Nelore bulls. 2012. 57 f. Tese (Doutorado em Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de) - Universidade Federal de Viçosa, Viçosa, 2012.http://locus.ufv.br/handle/123456789/1460The objectives of this study were to evaluate the potential of liquid watermelon extract as a natural diluent for cooling and/or freezing semen, and determining the cryoprotectant effect of liquid watermelon extract on the integrity of the plasma membrane of bovine sperm. Semen from four adult bulls were used, totalizing 10 semen samples for each one. After semen collection, the evaluation consisted of physical and morphological exam, live and dead cells (supravital test) and hypoosmotic swelling test. Afterwards, the fresh semen was divided in four equal parts and diluted with respective extenders: D1 (Bovimix® - control); D2 (Bovimix® base + 25 % of liquid watermelon extract); D3 (Bovimix® base + 50 % of liquid watermelon extract); and D4 (100 % of liquid watermelon extract base). After final dilutions, sperm motility and vigor were evaluated in each extender, and posterior seal. The straws were cooled and frozen in the freezing machine (Cryogen Dualflex®). Then, the straws were immersed in nitrogen. The thawing of the samples was done by immersion of the straws in a 37 °C water-bath for 30 minutes to evaluate the sperm motility and vigor, morphological characteristics of sperm, supravital test, hypoosmotic swelling test, epifluorescence test and thermoresistence test. The volume, motility, sperm vigor, sperm concentration per mL, supravital test, hypoosmotic swelling test, major and total sperm defects varied among animals, but showed no difference between them (P>0.05). The appearance, whirlpool and minor sperm defects showed differences among animals (P<0.05). Sperm motility showed high and positive correlation (r = 0.98) with the hypoosmotic swelling test. The supravital test showed high and positive correlation (r = 0.88) with the hypoosmotic swelling test. There was an average and negative correlation (r = -0.48) between motility and sperm concentration per mL, as well as average and negative correlation (r = -0.46) between the hypoosmotic swelling test and sperm concentration per mL. The motility and sperm vigor of the diluted semen (pre-cooling) in all extenders used, presented no difference between them (P>0.05). The motility upon thawing and at the end of thermoresistence test differed between extenders (P<0.05), where higher values were obtained by diluents D1 and D2, and lower values were obtained by diluents D3 and D4. At the beginning of the thermoresistence test, there was no difference between diluents D1 and D2. However, there was a difference when comparing the diluent D1 with D3 and D4. There was no difference between the diluent D2 and D3 or D2 and D4. At the end of the thermoresistence test, there was no difference between the diluents D1 and D2 or D1 and D4, but there was a difference between D1 and D3. There was no difference when comparing the diluent D2 with D3 and D4. The average sperm alive in the supravital test at the time of thawing in all extenders used, presented no difference between them (P>0.05). During the thermoresistence test, the average vigor of postthawed semen, in all extenders used, showed no difference between them (P>0.05). The hypoosmotic swelling test differed between extenders (P<0.05), where higher values were recorded in the diluent D1, an intermediate value was recorded in the diluent D2 and lower values were recorded in the diluents D3 and D4. There was no difference when comparing the diluent D1 with D2 and, no difference in between the diluents D2 and D3 or D2 and D4. However, there was a difference when comparing the diluent D1 with D3 and D4. There was no difference between the diluents D2 and D3 or D4. The epifluorescence test differed between extenders (P<0.05), but there was no difference between diluents D1 and D2. There was no difference between the diluents D2 and D3, or between the diluents D3 and D4. However, there was a difference when comparing D1 with D3 and D4. There were differences between diluents D2 and D4. The major and total sperm defects ranged among extenders, but showed no difference between them (P>0.05). However, minor sperm defects differed between extenders (P<0.05), where the highest values were observed in the diluent D3, and lower values were observed in the other diluents. There was no difference between diluent D1 and the other extenders. However, there was a difference between diluents D3 and D4. In the diluent D1, the values obtained in the supravital test showed average and positive correlation (r = 0.49) with sperm motility. The hypoosmotic swelling test showed high and positive correlation (r = 0.89) with intact sperm and, high and negative correlation (r = -0.80) with damaged spermatozoa observed in epifluorescence test. In the diluent D2, the values obtained in hypoosmotic test showed high and positive correlation (r = 0.86) with intact sperm, high and negative correlation (r = -0.83) with damaged spermatozoa and, average and positive correlation (r = 0.41) with semi-damaged sperm observed in epifluorescence test. In the diluent 3, the values obtained in the supravital test showed low and positive correlation (r = 0.33) with sperm motility. The hypoosmotic test showed high and positive correlation (r = 0.82) with sperm integrity, high and negative correlation (r = -0.80) with damaged sperm and, average and positive correlation (r = 0.51) with semi-damaged sperm observed in epifluorescence test. In the diluent D4, the values obtained in the supravital test showed average and positive correlation (r = 0.46) with sperm motility. The hypoosmotic swelling test showed high and positive correlation (r = 0.83) with intact sperm, high and negative correlation (r = - 0.85) with damaged sperm and, low and positive correlation (r = 0.38) with semidamaged sperm observed in epifluorescence test. At concentrations of 50 % and 100 %, the liquid watermelon extract was not effective in maintaining the integrity and sperm viability after thawing. At the concentration of 25 %, the liquid watermelon extract maintained the structural integrity of the membrane of spermatozoa during the cryopreservation process, and viable after the thermoresistence test, could be an alternative in the composition of diluents for cryopreservation of bovine semen.Os objetivos deste estudo foram avaliar o potencial do extrato líquido de melancia como meio diluidor natural no resfriamento e congelamento do sêmen, e determinar o efeito crioprotetor do extrato líquido de melancia sobre a integridade de membrana plasmática dos espermatozoides de bovinos. Foram utilizados quatro touros da raça Nelore. Para as coletas de sêmen, utilizou-se o método de eletroejaculação, onde se obteve 10 ejaculados por animal. Após a coleta, fez-se o exame físico do sêmen e morfológico dos espermatozoides, teste supravital e teste hiposmótico. Em seguida, o sêmen in natura foi dividido em quatro alíquotas iguais e diluído de acordo com os diluentes: D1 (Bovimix® - controle); D2 (base Bovimix® + 25 % de extrato líquido de melancia); D3 (base Bovimix® + 50 % de extrato líquido de melancia); e D4 (base 100 % de extrato líquido de melancia). Após as diluições finais, foram avaliados a motilidade e o vigor espermático de cada diluente, e posterior envase. As palhetas foram resfriadas e congeladas na máquina de congelamento (Cryogen Dualflex®). Após o congelamento, as palhetas foram imersas no nitrogênio líquido para o congelamento final do sêmen. As doses foram descongeladas em banho-maria a 37°C por 30 segundos, e acondicionadas em tubos plásticos de 1,5 mL e homogeneizadas para análise imediata de motilidade e vigor espermático, características morfológicas dos espermatozoides, teste supravital, teste hiposmótico, teste de epifluorescência e teste de termorresistência. No sêmen in natura, o volume, motilidade espermática progressiva, vigor espermático, concentração espermática por mL, teste supravital, teste hiposmótico, defeitos espermáticos maiores e totais variaram entre os animais, porém não houve diferença entre os mesmos (P>0,05). O aspecto, turbilhonamento e defeitos espermáticos menores apresentaram diferença entre os animais (P<0,05). A motilidade espermática apresentou correlação alta e positiva (r = 0,98) com o teste hiposmótico. O teste supravital apresentou correlação alta e positiva (r = 0,88) com o teste hiposmótico. Houve correlação média e negativa (r = - 0,48) entre a motilidade espermática progressiva e a concentração espermática por mL, bem como houve correlação média e negativa (r = -0,46) do teste hiposmótico com a concentração espermática por mL. No sêmen diluído (pré-resfriamento), as médias da motilidade e vigor espermático em todos os diluentes utilizados, não apresentaram diferença entre si (P>0,05). A motilidade espermática progressiva no momento do descongelamento e no final do TTR apresentou diferença entre os diluentes (P<0,05), onde os valores superiores foram obtidos pelos diluentes D1 e D2, e valores menores foram obtidos pelos diluentes D3 e D4. No início do TTR, não houve diferença do diluente D1 com o diluente D2. Porém, houve diferença do diluente D1 com os diluentes D3 e D4. Não houve diferença do diluente D2 com os diluentes D3 e D4. Enquanto que no final do TTR, não houve diferença do diluente D1 com os diluentes D2 e D4, porém, houve diferença do diluente D1 com o diluente D3. Não houve diferença do diluente D2 com os diluentes D3 e D4. As médias dos espermatozoides vivos no teste supravital no momento do descongelamento, em todos os diluentes utilizados, não apresentaram diferença entre si (P>0,05). Durante o TTR, as médias do vigor do sêmen pós-descongelado, em todos os diluentes utilizados, não apresentaram diferença entre si (P>0,05). O teste hiposmótico apresentou diferença entre os diluentes (P<0,05), onde valores mais altos foram registrados com o uso do diluente 1, valor intermediário foi registrado com o diluente D2 e valores inferiores foram registrados com os diluentes D3 e D4. Os valores do teste hiposmótico não diferiram entre os diluentes D1 e D2, assim como os valores do teste hiposmótico do diluente D2 não diferiu dos diluentes D3 e D4. Porém, os valores do teste hiposmótico do diluente D1 doram diferentes dos diluentes D3 e D4. Não houve diferença do diluente D2 com os diluentes D3 e D4. O teste de epifluorescência apresentou diferença entre os diluentes (P<0,05), porém não houve diferença do diluente D1 com o diluente D2. Não houve diferença entre os diluentes D2 e D3, bem como, não houve diferença entre os diluentes D3 e D4. Porém, houve diferença do diluente D1 com os diluentes D3 e D4. Houve diferença do diluente D2 com os diluentes D4. Os defeitos espermáticos maiores e totais variaram entre os diluentes, porém não houve diferença entre os mesmos (P>0,05). No entanto, os defeitos espermáticos menores apresentaram diferença entre os diluentes (P<0,05), onde o diluente D3 apresentou o maior número de espermatozoides com defeitos menores e os demais diluentes se apresentaram com valores inferiores. Não houve diferença do diluente D1 com os demais diluentes. Porém, houve diferença do diluente D3 com o diluente D4. No diluente 1, os valores obtidos no teste supravital apresentaram correlação média e positiva (r = 0,49) com a motilidade espermática. O teste hiposmótico apresentou correlação alta e positiva (r = 0,89) com os espermatozoides íntegros e correlação alta e negativa (r = -0,80) com os espermatozoides lesados observados no teste de epifluorescência. No diluente 2, os valores obtidos no teste hiposmótico apresentaram correlação alta e positiva (r = 0,86) com os espermatozoides íntegros, correlação alta e negativa (r = -0,83) com os espermatozoides lesados e correlação média e positiva (r = 0,41) com os espermatozoides semi-lesados observados no teste de epifluorescência. No diluente 3, os valores obtidos no teste supravital apresentaram correlação baixa e positiva (r = 0,33) com a motilidade espermática. O teste hiposmótico apresentou correlação alta e positiva (r = 0,82) com os espermatozoides íntegros, correlação alta e negativa (r = -0,80) com os espermatozoides lesados e correlação média e positiva (r = 0,51) com os espermatozoides semi-lesados observados no teste de epifluorescência. No diluente 4, os valores obtidos no teste supravital apresentaram correlação média e positiva (r = 0,46) com a motilidade espermática. O teste hiposmótico apresentou correlação alta e positiva (r = 0,83) com os espermatozoides íntegros e correlação alta e negativa (r = - 0,85) com os espermatozoides lesados e correlação baixa e positiva (r = 0,38) com os espermatozoides semi-lesados observados no teste de epifluorescência. Nas concentrações de 50 % e 100%, o extrato líquido de melancia não se mostrou eficaz na manutenção da integridade e viabilidade espermática pós-descongelamento. Na concentração de 25 %, o extrato líquido de melancia manteve a integridade estrutural da membrana dos espermatozoides durante o processo de criopreservação, bem como sua viabilidade após o teste de termorresistência, podendo ser uma alternativa na composição de diluentes para criopreservação de sêmen bovino.Coordenação de Aperfeiçoamento de Pessoal de Nível Superiorapplication/pdfporUniversidade Federal de ViçosaDoutorado em Medicina VeterináriaUFVBRBiotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. deAntioxidantesCrioprotetoresBovinosAntioxidantsCryoprotectorsCattleCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::REPRODUCAO ANIMALUso do extrato líquido de melancia (Citrullus lanatus) na criopreservação de sêmen de touros da raça NeloreUse of liquid watermelon extract (Citrullus lanatus) in cryopreservation of semen of Nelore bullsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALtexto completo.pdfapplication/pdf950785https://locus.ufv.br//bitstream/123456789/1460/1/texto%20completo.pdfd87988eac422eb97ddd4e0cec895d172MD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain123163https://locus.ufv.br//bitstream/123456789/1460/2/texto%20completo.pdf.txt660dd2210f83d8602e54843ca77437a9MD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3537https://locus.ufv.br//bitstream/123456789/1460/3/texto%20completo.pdf.jpg5bfc04514076e86df28a2a3502b24733MD53123456789/14602016-04-07 23:04:58.086oai:locus.ufv.br:123456789/1460Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-08T02:04:58LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false
dc.title.por.fl_str_mv Uso do extrato líquido de melancia (Citrullus lanatus) na criopreservação de sêmen de touros da raça Nelore
dc.title.alternative.eng.fl_str_mv Use of liquid watermelon extract (Citrullus lanatus) in cryopreservation of semen of Nelore bulls
title Uso do extrato líquido de melancia (Citrullus lanatus) na criopreservação de sêmen de touros da raça Nelore
spellingShingle Uso do extrato líquido de melancia (Citrullus lanatus) na criopreservação de sêmen de touros da raça Nelore
Castilho, Erick Fonseca de
Antioxidantes
Crioprotetores
Bovinos
Antioxidants
Cryoprotectors
Cattle
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::REPRODUCAO ANIMAL
title_short Uso do extrato líquido de melancia (Citrullus lanatus) na criopreservação de sêmen de touros da raça Nelore
title_full Uso do extrato líquido de melancia (Citrullus lanatus) na criopreservação de sêmen de touros da raça Nelore
title_fullStr Uso do extrato líquido de melancia (Citrullus lanatus) na criopreservação de sêmen de touros da raça Nelore
title_full_unstemmed Uso do extrato líquido de melancia (Citrullus lanatus) na criopreservação de sêmen de touros da raça Nelore
title_sort Uso do extrato líquido de melancia (Citrullus lanatus) na criopreservação de sêmen de touros da raça Nelore
author Castilho, Erick Fonseca de
author_facet Castilho, Erick Fonseca de
author_role author
dc.contributor.authorLattes.por.fl_str_mv http://lattes.cnpq.br/7525440848644647
dc.contributor.author.fl_str_mv Castilho, Erick Fonseca de
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 Fonseca, Jeferson Ferreira da
dc.contributor.advisor-co2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4703690H8
dc.contributor.advisor1.fl_str_mv Guimarães, José Domingos
dc.contributor.advisor1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782270U6
dc.contributor.referee1.fl_str_mv Siqueira, Jeanne Broch
dc.contributor.referee1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4766315A3
dc.contributor.referee2.fl_str_mv Paula, Tarcízio Antônio Rego de
dc.contributor.referee2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4701637D5
dc.contributor.referee3.fl_str_mv Guimarães, Simone Eliza Facioni
dc.contributor.referee3Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782526Y2
dc.contributor.referee4.fl_str_mv Torres, Ciro Alexandre Alves
dc.contributor.referee4Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787213D4
contributor_str_mv Costa, Eduardo Paulino da
Fonseca, Jeferson Ferreira da
Guimarães, José Domingos
Siqueira, Jeanne Broch
Paula, Tarcízio Antônio Rego de
Guimarães, Simone Eliza Facioni
Torres, Ciro Alexandre Alves
dc.subject.por.fl_str_mv Antioxidantes
Crioprotetores
Bovinos
topic Antioxidantes
Crioprotetores
Bovinos
Antioxidants
Cryoprotectors
Cattle
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::REPRODUCAO ANIMAL
dc.subject.eng.fl_str_mv Antioxidants
Cryoprotectors
Cattle
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::REPRODUCAO ANIMAL
description The objectives of this study were to evaluate the potential of liquid watermelon extract as a natural diluent for cooling and/or freezing semen, and determining the cryoprotectant effect of liquid watermelon extract on the integrity of the plasma membrane of bovine sperm. Semen from four adult bulls were used, totalizing 10 semen samples for each one. After semen collection, the evaluation consisted of physical and morphological exam, live and dead cells (supravital test) and hypoosmotic swelling test. Afterwards, the fresh semen was divided in four equal parts and diluted with respective extenders: D1 (Bovimix® - control); D2 (Bovimix® base + 25 % of liquid watermelon extract); D3 (Bovimix® base + 50 % of liquid watermelon extract); and D4 (100 % of liquid watermelon extract base). After final dilutions, sperm motility and vigor were evaluated in each extender, and posterior seal. The straws were cooled and frozen in the freezing machine (Cryogen Dualflex®). Then, the straws were immersed in nitrogen. The thawing of the samples was done by immersion of the straws in a 37 °C water-bath for 30 minutes to evaluate the sperm motility and vigor, morphological characteristics of sperm, supravital test, hypoosmotic swelling test, epifluorescence test and thermoresistence test. The volume, motility, sperm vigor, sperm concentration per mL, supravital test, hypoosmotic swelling test, major and total sperm defects varied among animals, but showed no difference between them (P>0.05). The appearance, whirlpool and minor sperm defects showed differences among animals (P<0.05). Sperm motility showed high and positive correlation (r = 0.98) with the hypoosmotic swelling test. The supravital test showed high and positive correlation (r = 0.88) with the hypoosmotic swelling test. There was an average and negative correlation (r = -0.48) between motility and sperm concentration per mL, as well as average and negative correlation (r = -0.46) between the hypoosmotic swelling test and sperm concentration per mL. The motility and sperm vigor of the diluted semen (pre-cooling) in all extenders used, presented no difference between them (P>0.05). The motility upon thawing and at the end of thermoresistence test differed between extenders (P<0.05), where higher values were obtained by diluents D1 and D2, and lower values were obtained by diluents D3 and D4. At the beginning of the thermoresistence test, there was no difference between diluents D1 and D2. However, there was a difference when comparing the diluent D1 with D3 and D4. There was no difference between the diluent D2 and D3 or D2 and D4. At the end of the thermoresistence test, there was no difference between the diluents D1 and D2 or D1 and D4, but there was a difference between D1 and D3. There was no difference when comparing the diluent D2 with D3 and D4. The average sperm alive in the supravital test at the time of thawing in all extenders used, presented no difference between them (P>0.05). During the thermoresistence test, the average vigor of postthawed semen, in all extenders used, showed no difference between them (P>0.05). The hypoosmotic swelling test differed between extenders (P<0.05), where higher values were recorded in the diluent D1, an intermediate value was recorded in the diluent D2 and lower values were recorded in the diluents D3 and D4. There was no difference when comparing the diluent D1 with D2 and, no difference in between the diluents D2 and D3 or D2 and D4. However, there was a difference when comparing the diluent D1 with D3 and D4. There was no difference between the diluents D2 and D3 or D4. The epifluorescence test differed between extenders (P<0.05), but there was no difference between diluents D1 and D2. There was no difference between the diluents D2 and D3, or between the diluents D3 and D4. However, there was a difference when comparing D1 with D3 and D4. There were differences between diluents D2 and D4. The major and total sperm defects ranged among extenders, but showed no difference between them (P>0.05). However, minor sperm defects differed between extenders (P<0.05), where the highest values were observed in the diluent D3, and lower values were observed in the other diluents. There was no difference between diluent D1 and the other extenders. However, there was a difference between diluents D3 and D4. In the diluent D1, the values obtained in the supravital test showed average and positive correlation (r = 0.49) with sperm motility. The hypoosmotic swelling test showed high and positive correlation (r = 0.89) with intact sperm and, high and negative correlation (r = -0.80) with damaged spermatozoa observed in epifluorescence test. In the diluent D2, the values obtained in hypoosmotic test showed high and positive correlation (r = 0.86) with intact sperm, high and negative correlation (r = -0.83) with damaged spermatozoa and, average and positive correlation (r = 0.41) with semi-damaged sperm observed in epifluorescence test. In the diluent 3, the values obtained in the supravital test showed low and positive correlation (r = 0.33) with sperm motility. The hypoosmotic test showed high and positive correlation (r = 0.82) with sperm integrity, high and negative correlation (r = -0.80) with damaged sperm and, average and positive correlation (r = 0.51) with semi-damaged sperm observed in epifluorescence test. In the diluent D4, the values obtained in the supravital test showed average and positive correlation (r = 0.46) with sperm motility. The hypoosmotic swelling test showed high and positive correlation (r = 0.83) with intact sperm, high and negative correlation (r = - 0.85) with damaged sperm and, low and positive correlation (r = 0.38) with semidamaged sperm observed in epifluorescence test. At concentrations of 50 % and 100 %, the liquid watermelon extract was not effective in maintaining the integrity and sperm viability after thawing. At the concentration of 25 %, the liquid watermelon extract maintained the structural integrity of the membrane of spermatozoa during the cryopreservation process, and viable after the thermoresistence test, could be an alternative in the composition of diluents for cryopreservation of bovine semen.
publishDate 2012
dc.date.issued.fl_str_mv 2012-05-28
dc.date.available.fl_str_mv 2013-11-29
2015-03-26T12:47:49Z
dc.date.accessioned.fl_str_mv 2015-03-26T12:47:49Z
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dc.identifier.citation.fl_str_mv CASTILHO, Erick Fonseca de. Use of liquid watermelon extract (Citrullus lanatus) in cryopreservation of semen of Nelore bulls. 2012. 57 f. Tese (Doutorado em Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de) - Universidade Federal de Viçosa, Viçosa, 2012.
dc.identifier.uri.fl_str_mv http://locus.ufv.br/handle/123456789/1460
identifier_str_mv CASTILHO, Erick Fonseca de. Use of liquid watermelon extract (Citrullus lanatus) in cryopreservation of semen of Nelore bulls. 2012. 57 f. Tese (Doutorado em Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de) - Universidade Federal de Viçosa, Viçosa, 2012.
url http://locus.ufv.br/handle/123456789/1460
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dc.publisher.none.fl_str_mv Universidade Federal de Viçosa
dc.publisher.program.fl_str_mv Doutorado em Medicina Veterinária
dc.publisher.initials.fl_str_mv 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
publisher.none.fl_str_mv Universidade Federal de Viçosa
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