EFFECT OF SEPHADEX SPERM FILTRATION AND ANTIOXIDANTS SUPPLEMENTATION ON POST THAW SEMEN QUALITY OF HOLSTEIN BULLS

Abstract

This study aimed to evaluate the efficacy of semen filtration using two different grades of Sephadex in enhancing the quality of low-grade ejaculates from Holstein bulls, in combination with antioxidant supplementation. Semen samples were collected from eight Holstein bulls over a period of eight weeks and subsequently divided into four experimental groups. The first group included SG5, SG6, and SG7, in which low-quality spermatozoa were filtered using Sephadex G-15. The second group consisted of SG8, SG9, and SG10, where low-quality spermatozoa were filtered using Sephadex G-25. A third group (SG1), comprising unfiltered low-quality semen, served as the negative control. Finally, a group of high-quality semen samples (SG2) was designated as the positive control. Groups SG3, SG6, and SG9 were supplemented with a combination of two amino acids (20 mM glycine and 1 mM glutamine), whereas groups SG4, SG7, and SG10 received a supplementation of 0.2 mM vitamin E and 0.8 mg/mL omega-3 fatty acids. Groups SG5 and SG8, which underwent filtration without any supplementation, served as control subgroups. The results demonstrated a significant improvement (P ≤ 0.01) in sperm motility, viability, normal morphology, acrosome integrity, and plasma membrane integrity in the groups filtered through Sephadex (SG5, SG6, SG7, SG8, SG9, and SG10) following both cooling and cryopreservation. Moreover, groups SG4, SG7, and SG10 showed a significant improvement (P ≤ 0.01) in sperm motility, viability, and plasma membrane integrity compared to the control groups SG8 and SG1. It can be concluded that sperm filtration is an effective method for reducing dead and abnormal sperm, as well as cellular debris, in low-quality ejaculates of Holstein bulls.