The impact of addition of ascorbic acid to cryopreservation medium on dog epididymal spermatozoa

Document Type : Original Article

Authors

1 Department of Theriogenology, Faculty of Veterinary Medicine, Cairo University, Giza 12211, Egypt

2 Animal Reproduction Research Institute, Giza, Egypt

Abstract

The objective of this study was to assess the effect of the addition of different concentrations of ascorbic acid to the extender on frozen-thawed epididymal dog spermatozoa. Epididymides from 17 castrated dogs were minced and incubated in a Tris buffer. The retrieved spermatozoa were diluted with Tris based-egg yolk-glycerol extender supplemented with different concentrations of ascorbic acid (0.45 mg/ml and 0.90 mg/ml) and the control (0.0 mg/ml). Diluted samples were equilibrated at 5°C for 2 h, packaged in 0.25 ml straws, and stored in liquid nitrogen (-196°C). After thawing (37°C for 30 s), sperm motility, viability, membrane integrity, acrosome integrity, DNA integrity, and lipid peroxidation by malondialdehyde (MDA) concentration were evaluated. The results were expressed as mean ± SE. Adding 0.90 mg/ml ascorbic acid to the cryopreservation medium significantly (P<0.05) improved motility, viability, membrane and acrosome integrity compared to the control. MDA concentration was significantly (P<0.05) reduced at 0.90 mg/ml related to the control. Percent of DNA damage was significantly (P<0.05) reduced in 0.45 mg/ml and 0.90 mg/ml ascorbic acid compared to the control. In conclusion, addition of ascorbic acid (0.90 mg/ml) to TCF extender resulted in a significant increase in the percentage of motility, viability, membrane intact, and acrosome- intact canine epididymal sperm, as well as the maintenance of DNA integrity and the reduction of lipid peroxidation at the membrane level.

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