Bicarbonato, albumina s�rica bovina e heparina afetam a integridade do acrossomo do espermatozoide bovino p�s-descongela��o? / Bicarbonate, bovine serum albumin and heparin affect acrosomal integrity of bovine post-thawing?

O trabalho teve como objetivo avaliar o efeito do bicarbonato, albumina s�rica bovina e heparina sobre a integridade do acrossomo do espermatozoide bovino criopreservado, durante incuba��o in vitro. Doses comerciais de s�men de dez touros foram submetidas �s an�lises de motilidade, vigor, integridade da membrana plasm�tica (eosina/nigrosina-EN) e status acrossomal(trypan blue/Giemsa-TBG), nos momentos p�s-descongela��o, p�s-wash e p�s-incuba��o por 5 horas a 37�C. Ap�s a descongela��o e lavagem por centrifuga��o em solu��o salina 0,9% (wash),as doses foram fracionadas e submetidas aos seguintes tratamentos: Talp-sp (controle); Talp-sp sem bicarbonato; Talp-sp sem albumina s�rica bovina e; Talp-sp acrescido de heparina. Os dados foram submetidos � an�lise de vari�ncia e as m�dias comparadas pelo teste de Duncan. Todas as caracter�sticas esperm�ticas sofreram efeito do tempo de incuba��o, diferindo significativamente dos valores obtidos p�s-descongela��o. Houve efeito significativo de tratamento para rea��o acrossomal verdadeira, com os maiores percentuais nos tratamentos controle (2,72�0,40) e com heparina (2,31�0,44), sendo que este �ltimo n�o diferiu dos demais. Houve intera��o significativa entre os momentos de avalia��o e os tratamentos para a vari�vel rea��o do acrossomo verdadeira, com os maiores valores nos tratamentos controle e com heparina (4,65�0,79 e 4,30�0,94%, respectivamente), n�o havendo diferen�a entre si. De acordo com os resultados obtidos conclui-se que a presen�a do bicarbonato ou da albumina s�rica bovina no meio de cultivo, leva ao aumento de espermatozoides apresentando rea��o do acrossomo verdadeira(AU)

The aim of this work was to evaluate de effect of bicarbonate, bovine serum albumin and heparin on sperm acrosome integrity of bovine cryopreserved semen, during in vitro incubation.Commercial doses from ten bulls were submitted to analysis of motility, vigour, plasmatic membrane integrity (eosin/nigrosin � EN) and acrosomal status (trypan blue/Giemsa), after thawing, wash and 5 hours incubation at 37�C. After thawing and washing by centrifugation with saline solution at 0.9%, doses were fractionated and submitted to following treatments:Talp-sp (control); Talp-sp bicarbonate free; Talp-sp bovine serum albumin free and; Talp-sp plus heparin. Data were submitted to ANOVA and means were compared by Duncan�s test. All sperm characteristics changed along incubation time with values differing significantly from those observed after thawing. Treatment had significant effect on the percentage of true acrosome reaction with the highest values being observed in control (2.72�0.40) and heparin(2.31�0.44) groups, this last being equal to the others. There was significant interaction between moments and treatments for true acrosome reaction variable with the highest values being observed in control and heparin groups (4.65�0.79 and 4.30�0.94%, respectively), although none difference had been found between these values. These results indicate that the presence of bicarbonate or bovine serum albumin in culture medium enhance the number of spermatozoa showing true acrosome reaction(AU)

Efeito do estresse cal�rico in vitro em fol�culos pr�-antrais de Bos indicus / Effect of in vitro heat stress in preantral follicles of Bos indicus

O presente estudo teve como objetivo verificar o efeito do estresse cal�rico na viabilidade morfologia nuclear de fol�culos pr�-antrais bovinos in vitro. Os ov�rios foram obtidos em batedouro e divididos em duas metades, uma para o controle (sem estresse cal�rico) e a outra aquecida a 42�C por 20 min em solu��o salina de NaCl a 0,9 %. Para an�lise da viabilidade, os fol�culos foram isolados mecanicamente e classificados em n�o vi�veis ou vi�veis quando foram corados ou n�o com o azul de tripan, respectivamente. Para an�lise da morfologia nuclear, as amostras foram fixadas em Carnoy e encaminhadas para rotina histol�gica cl�ssica. Diferen�a significativa (P<0,05) na viabilidade folicular foi observada entre o controle (76,8 %) e fol�culos submetidos aos estresse cal�rico (62,5 %). Em rela��o an�lise histol�gica no grupo controle, 87,2% apresentaram-se normais. No grupo exposto ao estresse cal�rico, todos os fol�culos analisados apresentaram altera��o na morfologia nuclear. Conclui-se que fol�culos pr�-antrais bovinos s�o sens�veis ao estresse cal�rico in vitro(AU)

The aim present was to evaluate the effect of heat stress in the viability and nuclear morphology of bovine preantral follicles in vitro. Ovaries were obtained from a slaughterhouse and were divided into two halves, one for control (no heat stress) and the other heated to 42�C for 20min in NaCl solution 0.9 %. For analysis of viability, the follicles were mechanically isolated and classified as non-viable or viable when they were stained or not with trypan blue, respectively. For analysis of nuclear morphology, the samples were fixed in Carnoy and sent for routine histology. Significant difference (P <0.05) in follicular viability was observed between the control (76.8 %) follicles and subjected to heat stress (62. 5%). For histological analysis in the control group, 87.2% were normal. In the group exposed to heat stress, all the follicles examined showed changes in nuclear morphology. Concluded that bovine preantral follicles are sensitive to heat stress in vitro(AU)

Efeito do estresse cal�rico in vitro em fol�culos pr�-antrais de Bos indicus / Effect of in vitro heat stress in preantral follicles of Bos indicus

O presente estudo teve como objetivo verificar o efeito do estresse cal�rico na viabilidade morfologia nuclear de fol�culos pr�-antrais bovinos in vitro. Os ov�rios foram obtidos em batedouro e divididos em duas metades, uma para o controle (sem estresse cal�rico) e a outra aquecida a 42�C por 20 min em solu��o salina de NaCl a 0,9 %. Para an�lise da viabilidade, os fol�culos foram isolados mecanicamente e classificados em n�o vi�veis ou vi�veis quando foram corados ou n�o com o azul de tripan, respectivamente. Para an�lise da morfologia nuclear, as amostras foram fixadas em Carnoy e encaminhadas para rotina histol�gica cl�ssica. Diferen�a significativa (P<0,05) na viabilidade folicular foi observada entre o controle (76,8 %) e fol�culos submetidos aos estresse cal�rico (62,5 %). Em rela��o an�lise histol�gica no grupo controle, 87,2% apresentaram-se normais. No grupo exposto ao estresse cal�rico, todos os fol�culos analisados apresentaram altera��o na morfologia nuclear. Conclui-se que fol�culos pr�-antrais bovinos s�o sens�veis ao estresse cal�rico in vitro

The aim present was to evaluate the effect of heat stress in the viability and nuclear morphology of bovine preantral follicles in vitro. Ovaries were obtained from a slaughterhouse and were divided into two halves, one for control (no heat stress) and the other heated to 42�C for 20min in NaCl solution 0.9 %. For analysis of viability, the follicles were mechanically isolated and classified as non-viable or viable when they were stained or not with trypan blue, respectively. For analysis of nuclear morphology, the samples were fixed in Carnoy and sent for routine histology. Significant difference (P <0.05) in follicular viability was observed between the control (76.8 %) follicles and subjected to heat stress (62. 5%). For histological analysis in the control group, 87.2% were normal. In the group exposed to heat stress, all the follicles examined showed changes in nuclear morphology. Concluded that bovine preantral follicles are sensitive to heat stress in vitro

Bicarbonato, albumina s�rica bovina e heparina afetam a integridade do acrossomo do espermatozoide bovino p�s-descongela��o? / Bicarbonate, bovine serum albumin and heparin affect acrosomal integrity of bovine post-thawing?

O trabalho teve como objetivo avaliar o efeito do bicarbonato, albumina s�rica bovina e heparina sobre a integridade do acrossomo do espermatozoide bovino criopreservado, durante incuba��o in vitro. Doses comerciais de s�men de dez touros foram submetidas �s an�lises de motilidade, vigor, integridade da membrana plasm�tica (eosina/nigrosina-EN) e status acrossomal(trypan blue/Giemsa-TBG), nos momentos p�s-descongela��o, p�s-wash e p�s-incuba��o por 5 horas a 37�C. Ap�s a descongela��o e lavagem por centrifuga��o em solu��o salina 0,9% (wash),as doses foram fracionadas e submetidas aos seguintes tratamentos: Talp-sp (controle); Talp-sp sem bicarbonato; Talp-sp sem albumina s�rica bovina e; Talp-sp acrescido de heparina. Os dados foram submetidos � an�lise de vari�ncia e as m�dias comparadas pelo teste de Duncan. Todas as caracter�sticas esperm�ticas sofreram efeito do tempo de incuba��o, diferindo significativamente dos valores obtidos p�s-descongela��o. Houve efeito significativo de tratamento para rea��o acrossomal verdadeira, com os maiores percentuais nos tratamentos controle (2,72�0,40) e com heparina (2,31�0,44), sendo que este �ltimo n�o diferiu dos demais. Houve intera��o significativa entre os momentos de avalia��o e os tratamentos para a vari�vel rea��o do acrossomo verdadeira, com os maiores valores nos tratamentos controle e com heparina (4,65�0,79 e 4,30�0,94%, respectivamente), n�o havendo diferen�a entre si. De acordo com os resultados obtidos conclui-se que a presen�a do bicarbonato ou da albumina s�rica bovina no meio de cultivo, leva ao aumento de espermatozoides apresentando rea��o do acrossomo verdadeira

The aim of this work was to evaluate de effect of bicarbonate, bovine serum albumin and heparin on sperm acrosome integrity of bovine cryopreserved semen, during in vitro incubation.Commercial doses from ten bulls were submitted to analysis of motility, vigour, plasmatic membrane integrity (eosin/nigrosin � EN) and acrosomal status (trypan blue/Giemsa), after thawing, wash and 5 hours incubation at 37�C. After thawing and washing by centrifugation with saline solution at 0.9%, doses were fractionated and submitted to following treatments:Talp-sp (control); Talp-sp bicarbonate free; Talp-sp bovine serum albumin free and; Talp-sp plus heparin. Data were submitted to ANOVA and means were compared by Duncan�s test. All sperm characteristics changed along incubation time with values differing significantly from those observed after thawing. Treatment had significant effect on the percentage of true acrosome reaction with the highest values being observed in control (2.72�0.40) and heparin(2.31�0.44) groups, this last being equal to the others. There was significant interaction between moments and treatments for true acrosome reaction variable with the highest values being observed in control and heparin groups (4.65�0.79 and 4.30�0.94%, respectively), although none difference had been found between these values. These results indicate that the presence of bicarbonate or bovine serum albumin in culture medium enhance the number of spermatozoa showing true acrosome reaction