Utilization of the osmotic shock to assess the frozen ram semen viability

  • Gabriel Felipe Oliveira de Menezes Universidade Federal de Sergipe, Aracaju, SE
  • Rodrigo Freitas Bittencourt Universidade Federal da Bahia, Salvador, BA
  • Antonio de Lisboa Ribeiro Filho Universidade Federal da Bahia, Salvador, BA
  • Marcus Chalhoub Universidade Federal da Bahia, Salvador, BA
  • Marta Freitas Bittencourt União Metropolitana para o Desenvolvimento da Educação e Cultura, Faculdade de Ciências Agrárias e da Saúde, Lauro André, BA
  • Eunice Oba Universidade Estadual Paulista Júlio de Mesquita Filho, Botucatu, SP
  • Sony Dimas Bicudo Universidade Estadual Paulista Júlio de Mesquita Filho, Botucatu, SP
Keywords: Ovine, Frozen semen, Hipoosmotic swelling test, Deionized water

Abstract

This study evaluated the use of hipoosmotic swelling test (HOST) with deionized water (0 mOsmol), as a method of post thaw ram semen evaluation and correlate their findings with different techniques of semen evaluation. Therefore, twenty semen samples of 20 different adult rams were assessed as for kinetic sperm parameters through computerized system (IVOS 12, Hamiton Thorn Biosciences, Beverly, MA, EUA) and subjective analysis. The sperm membranes viability was carried out by the association of fluorescent probes (propidium iodide, JC-1 and FITC-PSA). The structural integrity of the plasma membrane was also studied through supravital test with eosin and the functional integrity of membrane evaluated by doing the hipoosmotic swelling test with deionized water (0 mOsmol), in the following proportions: One part of semen for 10 (HOST 10), 50 (HOST 50) and 100 (HOST 100) parts of water. After semen dilution in the different proportions it was fixed in formalin-buffered saline and analyzed with regard to percentage of HOST reactive sperm (bent/coiled). The percentage of reaction obtained for HOST 10 (33,1%); HOST 50 (32,8%) and HOST 100 (31,8%) did not differ significantly. HOST 10 presented positive correlation with the plasma membrane integrity by the EOS (r = 0,80; p < 0,05). Positive correlations between HOST 50 and HOST 100 with sperm subpopulation with membrane integrity by fluorescence were observed (r = 0,83 and r = 0,85; p < 0,01). The findings suggest that the HOST with deionized water can provide additional information for post thawing ram sperm viability evaluation.

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Published
2013-10-29
How to Cite
Menezes, G. F., Bittencourt, R., Ribeiro Filho, A. de, Chalhoub, M., Bittencourt, M., Oba, E., & Bicudo, S. (2013). Utilization of the osmotic shock to assess the frozen ram semen viability. Brazilian Journal of Veterinary Research and Animal Science, 50(5), 396-405. https://doi.org/10.11606/issn.2318-3659.v50i5p396-405
Section
ARTICLES