Analysis of kinetic parameters of sexed Holstein-Friesian bull spermatozoa using Percoll density gradient centrifugation with computer-assisted sperm analysis

Vet World Vol.18 February-2025 Article - 5

Research Article

Veterinary World, 18(2): 287-295

https://doi.org/10.14202/vetworld.2025.287-295

Putri Utami¹, Aulia Puspita Anugra Yekti¹, Chairun Nisa Aperi Simbolon¹, Habib Asshidiq Syah¹, Anny Amaliya², Tri Agus Siswoyo³, Nurul Isnaini¹, and Trinil Susilawati¹

1. Department of Animal Reproduction and Breeding, Faculty of Animal Science, Universitas Brawijaya, Malang, Indonesia.
2. Singosari Artificial Insemination Center (SAIC), Malang, Indonesia.
3. Graduate Program of Biotechnology, the Center of Excellence on Crop Industrial Biotechnology (PUI-PT BioTIn), Universitas Jember, Jember, Indonesia.

Background and Aim: Artificial insemination (AI) is a key biotechnology for improving dairy cattle populations, offering genetic enhancement and increased milk production. The advent of sexed semen allows for the preferential selection of female offspring which is beneficial for dairy operations. This study aimed to evaluate and optimize a spermatozoa sexing method using Percoll density gradient centrifugation (PDGC) and analyze kinetic parameters of the separated spermatozoa using computer-assisted sperm analysis.

Materials and Methods: The study was conducted on two Holstein-Friesian bulls at the Singosari Artificial Insemination Center and Universitas Brawijaya, Indonesia. Semen samples underwent PDGC sexing at two density gradients, (T1) 20%-65% and (T2) 20%-60%. Kinetic parameters, including motility, velocity, and movement patterns, were assessed pre- and post-sexing. Statistical analyses were performed using a one-way analysis of variance and Duncan’s test to determine significant differences.

Results: Fresh semen (control) exhibited significantly higher motility (88.45%) compared to T1 (70.94%) and T2 (72.22%), with p < 0.01. Velocity parameters, including curvilinear velocity, were also significantly reduced post-sexing. However, motility levels in sexed samples still exceeded the 40% AI threshold. The 20%-65% gradient demonstrated better performance in maintaining sperm quality compared to the 20%-60% gradient.

Conclusion: Although sexing reduced motility and kinetic parameters, both gradients yielded semen suitable for AI applications. The 20%-65% gradient showed superior results, indicating its potential for optimizing the sexing process. Further research is recommended to refine the technique and improve the viability of sexed sperm.

Keywords: Artificial insemination, computer-assisted sperm analysis, Holstein-Friesian, Percoll density gradient centrifugation, sperm motility.

How to cite this article: Utami P, Yekti APA, Simbolon CNA, Syah HA, Amaliya A, Siswoyo TA, Isnaini N, and Susilawati T (2025) Analysis of kinetic parameters of sexed Holstein-Friesian bull spermatozoa using Percoll density gradient centrifugation with computer-assisted sperm analysis, Veterinary World, 18(2): 287–295.

Received: 2024-10-15 Accepted: 2025-01-07 Published online: 2025-02-13

Corresponding author: Trinil Susilawati E-mail: tsusilawati@ub.ac.id

DOI: 10.14202/vetworld.2025.287-295

Copyright: Utami, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/ by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/ publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.