Enhanced sperm sexing efficiency and quality preservation in Bali bulls using freeze-dried albumin separation media

Abstract

Background: Sperm sexing is a crucial technique in livestock breeding, allowing for the selection of offspring of the desired sex. Traditional methods present challenges, including high costs, time consumption, and potential damage to sperm quality. This study explored an alternative approach using freeze-dried albumin as a medium for sperm separation in Bali bulls, a breed of significant economic importance in Indonesia. Aim: This study aimed to compare the advantages of freeze-dried albumin and fresh albumin for the separation of Bali bull sperm based on the existing albumin gradient separation technique. The evaluation focused on the impact of sperm separation, quality preservation, and kinematic properties. In addition, this study seeks to understand the broader applicability of this method beyond Bali bulls, considering its potential integration into various livestock reproductive programs. Methods: Fresh semen from Bali bulls was processed using two different sperm separation media: fresh albumin and freeze-dried albumin. Sperm motility, viability, membrane integrity, acrosome integrity, and kinematic parameters (e.g., velocity and linearity) were measured and compared between the two mediums. The proportions of X- and Y-chromosome-bearing sperm in the separated fractions were also assessed. Results: The study found no significant differences in sperm quality parameters (motility, viability, abnormalities, membrane integrity, and acrosome integrity.) between the two mediums. In contrast, freeze-dried albumin resulted in higher separation efficiency, with a greater proportion of X-bearing sperm in the upper gradient layer, compared with fresh albumin. Kinematic analysis showed a slight increase of about 2%-4% inmotility and stability in sperm treated with freeze-dried albumin. Conclusion: Freeze-dried albumin is a viable alternative to fresh albumin for sperm sexing because it improves separation efficiency while maintaining sperm quality. The advantages of stability, ease of storage, and cost- effectiveness make it an ideal choice for resource-limited settings, potentially advancing reproductive biotechnology in livestock breeding.

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