Abstract
Soybean lecithin (SL) has been used as a substitute for egg yolk (EY) in stallion semen cryopreservation, but its mechanism to maintain the integrity of the sperm plasma membrane is not clear. In this experiment, we compared effects of different SL concentrations (10%, 20%, 30% and 40%, v/v) on physiological characteristics of stallion sperm after freezing and thawing in the extender with or without EY. Semen was collected from three fertile stallions, each of them was collected four times. After collection, semen was diluted 1:1 with milk-based extender (INRA82) and split into two aliquots and centrifuged at 600 g for 10 min. After removal of the supernatant, sperm pellets were resuspended with different freezing extenders. Semen was packaged in 0.25mL straws and frozen in liquid nitrogen (LN2) vapor (5 cm above LN2 surface) for 15 min, then plunged into LN2 for storage. Straws were stored in LN2 for one month, and frozen semen was thawed in a water bath at 37 C for 30s. Sperm motility parameters were evaluated by computer analysis system (CASA), and sperm plasma membrane integrity was detected by the hypoosmotic swelling test (HOST). Sperm mitochondrial membrane potential was checked by flow cytometry after fluorescence labeling with JC-1 and PI, and lipid oxidative production was detected by the generation of malondialdehyde (MDA). All
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