Quality and Kinematic Characteristics of Frozen Sexed Bali Bull Sperm: Optimizing Percoll Density Gradient Centrifugation
Quality and Kinematic Characteristics of Frozen Sexed Bali Bull Sperm: Optimizing Percoll Density Gradient Centrifugation
Nanda Ayu Rahmawati1, Aulia Puspita Anugra Yekti1, Aditiya Wahyudi1, Putri Utami1, Habib Asshidiq Syah1, Fardha Ad Durrun Nafis2, Sri Wahjuningsih1, Achadiah Rachmawati1, Nurul Isnaini1, Trinil Susilawati1*
ABSTRACT
This research aims to determine and evaluate the quality, proportion and kinematic characteristics of frozen Bali bull sexed sperm using Percoll density gradient centrifugation (PDGC) method with various gradients using egg yolk tris aminomethane diluent. Semen was obtained from a Bali bull, with a confirmed individual motility of ≥ 70%, for this research. The data were analyzed by Analysis of Variance (ANOVA) with a Randomized Group Design in R studio (4.4.3 version). T1 = ten gradients, T2 = five gradients, and T3 = three gradients, each consisting of ten replicates. The analysis showed that gradient variation significantly affected individual motility in the bottom layer of sexed semen before freezing (P<0.01). It also had a very significant impact on individual motility, concentration, and total motile sperm (TMS) in the bottom layer of frozen sexed semen (P<0.01) and a significant effect on the viability of the top layer of frozen sexed semen (P<0.05). Meanwhile, gradient variation did not affect kinematic characteristic parameters with CASA (P>0.05). Regarding the proportion of X and Y sexed sperm, the results were: T1 = 85.40% and 79.80%; T2 = 81.20% and 77.90%; T3 = 75.10% and 79.90%. In conclusion, the five-gradients Percoll (bottom layer) can replace the ten-gradients PDGC method based on visual motility and proportion. Further research using 1 mL fresh semen with five and three gradients is recommended for improved results.
Keywords | Bali bull, Computer-assisted sperm analysis, Freezing, Percoll density gradient centrifugation, Proportion, Sexed semen
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July 2025
Adv. Anim. Vet. Sci., Vol. 13, Iss. 7,
