Abstract: In order to screen suitable low temperature preservation conditions and diluent formulation for donkey sperm, the low temperature preservation effects of the filtration methods for diluent and adding different concentrations of Vitamin C（VC）, Vitamin E（VE）, graphene oxide on donkey sperm were studied. The results showed that the better effect of low temperature preservation was obtained by adding 15% yolk after filtration of base solution than the diluent filtered after adding 15% yolk to the base solution or the yolk was filtered and then added to the diluent in the base solution by 15% volume percentage. The sperm motility still exceeded 0.30 after 8 days of preservation with diluents of 200 µg/100 mL and 300 µg/100 mL VC, significantly higher than that with diluents of 100 µg/100 mL VC（P<0.05）. When low temperature preservation for 10 days, adding 5% VE diluent had a good effect, and the sperm motility was still close to 0.30, which was significantly better than adding 10% and 15% VE diluents（P<0.05）. When low temperature preservation for 8 days, adding 30% graphene oxide diluent had a good effect, and the sperm motility was still close to 0.30, which was significantly better than adding 20% and 25% graphene oxide diluents（P<0.05）. In conclusion, the good effect was detected for donkey sperm low temperature preservation by the filtration method of adding yolk after filtration of base liquid, adding 200 µg/100 mL VC or 300 µg/100 mL VC, adding 5% VE, and adding 30% graphene oxide in diluent.

Key words: donkey, sperm, low temperature preservation, graphene oxide, VC, VE