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The aim of the present study was to elucidate the mode of action of goat bulbourethral lipase (BUSgp60 lipase) previously identified as responsible for the deterioration of goat sperm viability in skimmed milk-based extenders. Milk fractions were purified by micro- and ultrafiltration and characterized by SDS-PAGE, thin layer chromatography, triglyceride quantitative analysis and by their ability to potentiate the lipase and the sperm-deteriorating activity of the bulbourethral lipase. Components in both the phosphocaseinate and soluble whey protein fractions enhanced the lipase activity of BUSgp60 but only the phosphocaseinate fraction, which contains triglycerides, promoted deterioration of spermatozoa in the presence of bulbourethral gland secretion. These data suggest that the sperm-deteriorating effect of bulbourethral gland secretion is due to the catalysis of triglyceride hydrolysis, and that proteins increase this activity. BUSgp60 hydrolysed milk triglycerides and triolein very effectively, and its lipase activity was enhanced by several highly purified milk proteins. The major cis-unsaturated fatty acid from milk (oleic acid) but not the major saturated fatty acid (palmitic acid) exhibited dose-dependent detrimental effects on goat spermatozoa. Therefore, the catalysis of oleic acid formation from residual milk triglycerides by BUSgp60 appears responsible for the deterioration of goat spermatozoa when unwashed semen is diluted in skimmed milk-based extenders. The precise mechanism of action of oleic acid remains to be elucidated but the drawbacks of washing buck semen might be avoided by inhibiting BUSgp60 or by depriving it of substrate.

Deterioration of goat spermatozoa in skimmed milk-based extenders as a result of oleic acid released by the bulbourethral lipase BUSgp60