Quality of Sexed Bali Bull Semen Post-Thawing Using Percoll Density Gradient Centrifugation with Different Gradients and AndroMed® Diluent

Sexing is a reproduction technique used to modify the natural 50:50 spermatozoa ratio through specific methods to achieve a desired proportion, enabling the production of calves with the preferred sex. This study is the first to systematically investigate the use of three simplified levels of Percoll Density Gradient Centrifugation (PDGC) combined with Andromed® extender for improving both the efficiency of X-and Y-spermatozoa separation and the post-thaw quality of Bali bull semen. This dual-focused approach aims to enhance sex-specific sperm sorting while preserving key functional attributes of cryopreserved sperm, including motility, viability, and membrane integrity. This study evaluated the quality of sexed frozen semen produced using the Percoll Density Gradient Centrifugation (PDGC) method with three different gradient levels: 10, 5, and 3. Semen collection from bull was performed using an artificial vagina. The experimental treatments included T1: 10 gradients, T2: 5 gradients, and T3: 3 gradients, each with 10 replications. Statistical analysis revealed there were no significant differences (P>0.05) observed in individual motility, viability, abnormality, concentration, total motile spermatozoa, motility, progressive motility, and kinematic parameters, including VCL, VSL, VAP, LIN, STR, WOB, ALH, BCF, DAP, DSL, and DCL. In conclusion, the different gradient levels used in the sexing process did not significantly affect the quality of frozen sexed semen processed with Andromed® diluent. However, the use of 3 gradients is recommended, as it demonstrated higher values for individual motility of sexed spermatozoa.