Evaluation of sperm sexing technique using BSA column medium in Garut rams: Enhancing sperm quality, kinematics, and fertility performance for improved breeding efficiency.

Background: Sperm sexing techniques face challenges such as damage to sperm, lower fertility rates, high costs, and technical limitations, which hinder their widespread application... This study evaluates the effectiveness of BSA and albumin column sperm sexing techniques in Garut rams, focusing on separation efficiency, sperm quality, and fertility post-AI. Evaluating alternative methods like the BSA column medium in Garut rams could offers critical insights into the practical application of sexing methods, potentially improving the viability and practicality of sperm sexing in local Indonesian livestock.. Aim: This study aims to evaluated two sperm sexing techniques-Bovine Serum Albumin (BSA) column and albumin column-in Garut rams (Ovis aries), an economically important breed in Indonesia. The research focused on sperm separation efficiency, sperm quality, and fertility performance following artificial insemination (AI). Methods: This study compared BSA and albumin column techniques for sperm sexing in Garut rams. Sperm quality, kinematics, and sexing validation were analyzed using CASA and morphometric analysis. Frozen sexed semen was used for artificial insemination, and pregnancy rates were recorded. Data were analyzed using the Mann-Whitney U test and Pearson correlation to assess differences and relationships among all parameters, with significance set at p < 0.05. Results: BSA medium outperforms Albumin in motility, kinematic stability (Y-sperm), and fertility, though abnormalities and movement patterns (e.g., WOB) require optimization. Pre-sexing Garut ram sperm averaged 70.4% motility, 78.4% viability, and 7.8% abnormalities. Y-sexed sperm in BSA medium showed higher motility than in Albumin, while viability remained stable (74-75%); X-sexed sperm in Albumin had the highest abnormalities (11.2%). Post-sexing, VSL and DCL were significantly higher for Y-sperm in BSA (*p* < 0.05), but other parameters (VCL, VAP) showed no difference. Sperm post-sexing exhibited STR (90.06%) > WOB (81.88%) > LIN (72.72%), with WOB differing significantly for X-sperm between media (p < 0.05). BSA yielded higher fractions of both X- (65.4% vs. 62.8%) and Y-sperm (66.4% vs. 61.6%) than Albumin (p< 0.05). Fertility rates were higher in BSA for X- (62.8% vs. 56.6%) and Y-sperm (63.4% vs. 57.8%) (p< 0.05). Conclusion: The BSA column method is a cost-effective alternative to flow cytometry for sperm sexing in Garut rams, preserving sperm quality and improving fertility. The BSA column method provides small-scale breeders with an affordable and practical approach to sperm sexing, offering reasonable gender accuracy. However, its adoption requires careful consideration of potential trade-offs in sperm quality and fertility rates. While the technique's simplicity is advantageous, optimal outcomes depend on rigorous sperm quality management and possible supplementation with alternative albumin sources.