Abstract
In this study, the cryopreservation methods for Bama miniature pig semen were investigated and optimized. First, using an orthogonal experimental design, the semen freezing-thawing procedure for Bama miniature boars was optimized based on analysis of the effects of concentrations of LDL (LC, parameter A), trehalose (TC, parameter B) and glycerol (GC, parameter C), the equilibration time at 15°C (ET, parameter D), and the thawing method (TM, parameter E) on sperm motility. The results showed that the effects of the parameters could be arranged as A>C>B>D>E. The LDL concentration and final glycerol concentration had exceedingly significant effects on the motility of thawed spermatozoa (P<0.01), and the effects of the trehalose concentration, equilibration time at 15°C, and the thawing method were not significant (P>0.05). Scheme 2 (A3B4C2D3E1) gave a motility of 52.26% after thawing. Then, using sperm motility, acrosome integrity, plasma membrane integrity, and DNA injury rate as indicators, four combinations, on the basis of scheme 2, were designed to analyze the protective effects of different combinations of LDL, glycerol, and trehalose; the results showed that combination of 9% LDL, 200 mM trehalose, and 2% glycerol (i.e., combination 4) demonstrated significantly better protective effects than the other combinations (P<0.05), further verifying that scheme 2 was the best for cryopreservation of Bama miniature boar semen. In this way, a method with favorable performance was established for cryopreservation of semen of Bama miniature boars.
