Effect of tris-citric acid-egg yolk-glycerol and vitrified medium with and without ascorbic acid on the quality of post- thaw canine semen

Elbehiry, Mohamed A.; Adel, Mohamed; Hattab, Sayed; Ghallab, Rezk; Sobhy, Kamel

doi:10.21608/djvs.2022.169032.1098

Effect of tris-citric acid-egg yolk-glycerol and vitrified medium with and without ascorbic acid on the quality of post- thaw canine semen

Document Type : Original Article

Authors

¹ Department of Theriogenology, Faculty of Veterinary Medicine, Damanhour University, Damanhour, Egypt

² Departments of Theriogenology, Faculty of Veterinary Medicine, Alexandria University, Alexandria, Egypt

³ Department of Theriogenology, Faculty of Veterinary Medicine Matrouh University, Egypt

10.21608/djvs.2022.169032.1098

Abstract

The study aimed to compare vitrified medium (Soya lecithin 1% and 0.25 M sucrose) with tris-citric acid fructose-egg yolk 20%-glycerol3% (TCF-EY-GL) for freezing of canine semen and assess the impact of adding ascorbic acid to both extenders on the post-thawing semen quality. Semen was collected from 3 healthy and fertile German shepherd dogs by digital manipulation and stimulation of the bulbus glandis. Collected semen was initially evaluated by a computerized assisted semen analyzer (CASA). Ejaculates from the dogs were pooled and divided into 4 equal parts and diluted 1:1 with the following 4 extenders: TCF-EY-GL with ascorbic acid (Ex1), TCF-EY-GL without ascorbic acid (Ex2), vitrified medium with ascorbic acid (Ex3) and vitrified medium without ascorbic acid (Ex4). The extended semen allowed for equilibrating for 1 h in a refrigerator set at 4 ºC then packed into 0.5ml straws. Straws containing semen diluted in Ex1 and2 were frozen by the conventional method however, those containing semen diluted in Ex3 and Ex4 (vitrified media) were plunged directly in liquid nitrogen. Semen samples were evaluated by CASA immediately after thawing. Results revealed that progressive motility, normal morphology, and acrosome integrity were significantly higher in vitrified medium (Soya lecithin 1% and 0.25 M sucrose) than those obtained in TCF-EY-glycerol however, plasma membrane integrity was significantly improved in TCF-EY-GL compared with that observed in the vitrified medium. The addition of ascorbic acids to vitrified medium and TCF-EY-GL resulted in a significant increase in all semen quality parameters. Total sperm motility, progressive motility, and normal sperm morphology were significantly higher in vitrified medium with ascorbic acid than those observed in TCF-EY-GL with ascorbic acid. It was concluded that the addition of ascorbic acid to vitrified medium or TCF-EY-GL extender resulted in significant improvement in post-thawing canine semen quality and the vitrified medium can be used as a good alternative for TCF-EY-GL extender to facilitate freezing and avoid undesirable use of EY.

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