Study On The Application Effect Of Four Kinds Of Antioxidants In The Freezing Perservation Of Donkey Semen

Guanzhong donkey is a unique variety of Shaanxi. At present, it has become less and less.It is very important to protect it. Application of semen freezing technology can contribute to the protection of the Guanzhong donkey`s species. Finding the optimal cryoprotectants was still one of the main and important problems at present. Vitamin C, Vitamin E, GSH and Lamarin are commonly used antioxidant. This research used semen samples from Guanzhong donkey as the investigation target, to explore the effects of several antioxidants on donkey cryo-sperm. The sperm quality was evaluated by analyzing thawed donkey permatozoa motility, viability, mitochondrial membrane integrity, plasma membrane integrity and acrosomal integrity. The experimental techniques of eosin staining, hypoosmotic swelling test(HOST), propidium iodide(PI) and rhodamin 123(Rh 123) staining, and fluorescein isothiocyanate-peanut agglutinin(FITC-PNA) staining were used to test sperm viability, plasma membrane integrity, mitochondrial membrane integrity and acrosomal integrity. The results of this research were shown below:1.The group of II is the best group in this text. The indicators are better than the other groups.2. Certain concentration of both vitamin C and vitamin E added separately to the basic freezing diluent for donkey spermatozoa could effectively improve the sperm viability, sperm acrosome integrity rate, plasma membrane integrity rate and mitochondrial activity of donkey spermatozoa after frozen-thawed. The best results were obtained by the group adding 400 mg/L vitamin and the group adding 600 mg/L vitamin E. Compared with the control group, the difference was significant(P<0.05)3. Compared with adding individual, combination of the two applications could significantly improve the sperm viability, sperm acrosome integrity rate, plasma membrane integrity rate and mitochondrial activity of donkey spermatozoa after frozen-thawed. The concentrations of the best compatibility were 200 mg / L vitamin and 200 mg / L vitamin E(P <0.05).4.Certain concentration of both GSH and lamarin added separately to the basic freezing diluent for donkey spermatozoa could effectively improve the sperm viability, sperm acrosome integrity rate, plasma membrane integrity rate and mitochondrial activity of donkey spermatozoa after frozen-thawed. The best results were obtained by the group adding 1000 mg/L GSH and the group adding 1500 mg/L lamarin. Compared with the control group, the difference was significant(P<0.05).5. Compared with adding individual, combination of the two applications could signifi-cantly improve the sperm viability, sperm acrosome integrity rate, plasma membrane integrity rate and mitochondrial activity of donkey spermatozoa after frozen-thawed. The concentrations of the best compatibility were 500 mg / L GSH and 1500 mg / L lamarin(P<0.05).