Post-thawed semen quality and genomic variation in indigenous Indonesian buffalo breeds
Budi Utomo1*, Rimayanti Rimayanti1, Sri Mulyati1, Tjuk Imam Restiadi1, Nurul Khaeriyah Amrullah2, Faheem Ahmed Khan3, Syahruddin Said4, Santoso Santoso5, Athhar Manabi Diansyah6, Muhammad Rizal7, Rahmat Rahmat7, Muhammad Fajar Amrullah8, Fuad Hasan9, Aeni Nurlatifah10
1Division of Veterinary Reproduction, Faculty of Veterinary Medicine, Universitas Airlangga, City of Surabaya, East Java, Indonesia
2Bachelor program of Veterinary Medicine, Faculty of Veterinary Medicine, Universitas Airlangga, City of Surabaya, East Java, Indonesia
3Department of Transfusion Medicine and Clinical Microbiology, Faculty of Allied Health Sciences, Chulalongkorn University, Bangkok, Thailand
4Research Center for Applied Zoology, National Research and Innovation Agency, Cibinong Science Center, Jl. Raya Jakarta, Bogor, West Java, Indonesia
5Research Center for Animal Husbandry, National Research and Innovation Agency, Cibinong Science Center, Jl. Raya Jakarta, Bogor, West Java, Indonesia
6Department of Animal Production, Faculty of Animal Science, Hasanuddin University, Jl. Perintis Kemerdekaan Km. 10 Tamalanrea Makassar, South Sulawesi, Indonesia
7Faculty of Agriculture, Lambung Mangkurat University, Jl. Jenderal Ahmad Yani Km. 36 Banjarbaru, South Kalimantan, Indonesia
8Doctoral Program of Animal Biomedical Science, School of Veterinary Medicine and Biomedical Science, Bogor Agricultural University, Jl. Agatis, Kampus IPB Darmaga Bogor, Indonesia
9Animal Science Study Program, Faculty ofAgriiculture, Universitas Sumatera Utara, Medan 20155, Indonesia
10Department of Nutrition and Feed Technology, Faculty of Animal Science, Gadjah Mada University, Depok, Sleman Regency, Special Region of Yogyakarta, Indonesia
Received:13 August2025/ Revised:11 October 2025/ Accepted:14 October 2025/ Published Online: 21 October 2025
Abstract
Buffaloes (Bubalus bubalis) play a vital role in Indonesian livestock systems, but artificial insemination (AI) programs are often hindered by the decline in semen quality after cryopreservation. This study aimed to evaluate post-thawed semen performance and genetic variability among Toraja, Kalsel, and Silangit buffalo bulls, with the goal of identifying phenotypic and molecular traits linked to cryotolerance. A total of 60 frozen semen straws (20 per bull) were assessed for motility, viability, membrane integrity, and acrosome integrity using computer-assisted semen analysis (CASA) and functional assays. Genomic DNA obtained from blood samples was subjected to PCR amplification of candidate fertility genes (OPN, IGF-1, LHβ, SPAG11B, and TNP1), followed by sequence analysis to identify nucleotide mutations and deduced amino acid substitutions. Results showed that Silangit bulls had significantly greater viability (81.49%) and membrane integrity (89.70%) than Toraja and Kalsel bulls (p < 0.001), suggesting superior tolerance to freezing. All breeds displayed conserved G+C content (55–62%), but mutation patterns varied: OPN mutations appeared only in Toraja bulls, while LHβ and TNP1 showed higher mutation frequencies in Silangit bulls. Correlation tests revealed positive associations between LHβ mutations and sperm viability/straightness (r = 0.999–1.000, p < 0.05), while variations in OPN and SPAG11B were negatively linked to motility, acrosome integrity, and survival. These findings indicate that semen cryotolerance differences among buffalo breeds are influenced by specific genetic variations. Despite a limited sample size and narrow genomic coverage, the study highlights the value of integrating semen quality analysis with genomic tools for sire selection and improving AI success in native buffalo populations.
