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Enzyme leakage during cryopreservation of ram spermatozoa
Affiliation:1. School of Life Sciences, Sun Yat-sen University, Guangzhou 510006, China;2. Guangdong Provincial Key Laboratory for Aquatic Economic Animals, Institute of Aquatic Economic Animals, School of Life Sciences, Sun Yat-sen University, Guangzhou 510006, China;3. Guangdong Provincial Engineering Technology, Research Center for Healthy Breeding of Important Economic Fish, Guangzhou 510006, China
Abstract:Spermatozoal motility and enzyme leakage during cryopreservation of ram spermatozoa was evaluated in the presence and absence of antifreeze proteins (AFP). Loss of spermatozoal motility due to the cryopreservation process was reduced by 10% in the presence of AFP. Samples of the diluted semen at various stages of the cryopreservation process were assayed for aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), acid phosphatase (ACP) and lactate dehydrogenase (LDH). The levels of AST, ALT and ACP were low in ram spermatozoa and were not considered suitable for the objective measurement of spermatozoal damage. ALP leaked during the cooling and freezing process, whereas LDH leakage was prevalent during cooling, freezing and post-thaw incubation of spermatozoa. Changes in ALP and LDH could be used as marker enzymes in the development of semen processing protocols and of semen diluents.
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