Influence of alpha linolenic acid on the motility, viability, antioxidant activity and fertility of frozen-thawed New Zealand white rabbit buck semen

Abstract

Freezing and thawing processes result in production and accumulation of high concentrations of reactive oxygen species that are detrimental to spermatozoal motility and fertility. Therefore, supplementation of exogenous source of antioxidants to freezing diluent is crucial. The aim of the present study was to investigate for the first time whether supplementation of semen diluent with alpha linolenic acid (ALA) can improve motility, viability, membrane integrity, antioxidant status and fertility of post-thaw rabbit spermatozoa. Semen was collected and pooled from fifteen New Zealand white rabbit bucks. Semen samples were diluted with a tris-citrate-glucose (TCG) extender supplemented with ALA (0, 50, 75 and 100 μmol). Then, extended rabbit semen was cooled at 5°C and cryopreserved in liquid nitrogen. After thawing, spermatozoal quality parameters (individual motility %, viability %, osmotic resistance %, and acrosome integrity %), antioxidant activity (SOD, CAT, and GSH activities), lipid peroxidation (malondialdehyde) and fertility (conception and kindling rates) were evaluated. Results revealed that supplementation of rabbit semen extender with 50 μmol ALA significantly (P<0.05) increased spermatozoal characteristics including motility (56.54%), viability (60.01%), acrosome status (72.66%) and membrane integrity (59.13%). The activity of semen antioxidant enzymes (SOD, CAT, and GSH) showed a significant improvement with a marked decrease in lipid peroxidation. Moreover, the conception (73.30%) and kindling (70.00%) rates were significantly (P<0.05) higher in does inseminated with thawed semen treated with 50 μmol ALA in comparison with other concentrations (0, 75 and 100 μmol). In summary, supplementation of rabbit semen extender with 50 μmol ALA improved motility, viability, membrane integrity, acrosome integrity, antioxidant enzymes activity and fertility of post-thaw rabbit spermatozoa. Our findings suggested that higher concentrations of ALA are detrimental to post-thaw characteristics of New Zealand white rabbit buck spermatozoa. To achieve better results, the semen freezing extender should be supplemented with ALA at lower concentrations, especially 50 μmol.