Cryopreservation of Bali Bull Semen in CEP Diluent with the Addition of Various Antioxidants

¹ Animal Bioscience Department, Faculty of Food Security, Universitas Negeri Surabaya, Indonesia
² Aquaculture Department, Faculty of Food Security, Universitas Negeri Surabaya, Indonesia
³ Biology Department, Faculty of Mathematics and Natural Science, Universitas Negeri Surabaya, Indonesia
⁴ Singosari National Artificial Insemination Center, Malang, Indonesia

^* Corresponding author: This email address is being protected from spambots. You need JavaScript enabled to view it.

Abstract

Cryopreservation of semen was a crucial technology in livestock breeding; however, it compromised sperm quality due to oxidative stress, lipid peroxidation, and structural damage. This study aimed to determine the effect of adding α-tocopherol and glutathione antioxidants in Caudal Epididymal Plasma (CEP) diluent on the quality of frozen Bali bull semen. Four groups of CEP semen diluents were prepared with different antioxidant supplementation: P1 (CEP), P2 (CEP + 2 mM α-tocopherol), P3 (CEP + 0.75 mM glutathione), and P4 (CEP + 2 mM α-tocopherol + 0.75 mM glutathione), while Tris Egg Yolk extender was used as a comparator (P5). Semen collected from two Bali bulls was evaluated before and after the freezing and thawing processes using parameters such as motility, viability, membrane integrity, lipid peroxidation (MDA), and DNA integrity. The results showed that the P2 group supplemented with 2 mM α-tocopherol had the highest post-thawed motility, viability, and membrane integrity compared to other groups (P<0.05). The addition of α-tocopherol and glutathione proved to reduce MDA concentrations and improve the quality of frozen semen. This study concluded that α-tocopherol and glutathione improved the quality of Bali bull semen after cryopreservation, with α-tocopherol showing more significant results.