Sahiruddin Sahiruddin1*, Muhammad Yusuf1, Athhar Manabi Diansyah1, Masturi Masturi1, Herdis Herdis2, Tulus Maulana3, Syaputra Wibowo4
1Faculty of Animal Science, Hasanuddin University, Jl. Perintis Kemerdekaan Km. 10 Tamalanrea Makassar, South Sulawesi 90245, Indonesia
2Research Center for Animal Husbandry, National Research and Innovation Agency, Cibinong Science Center, Jl. Raya Jakarta, Bogor 16915, West Java, Indonesia
3Research Center for Applied Zoology, National Research and Innovation Agency, Cibinong Science Center, Jl. Raya Jakarta, Bogor 16915, West Java, Indonesia
4Eijkman Research Center for Molecular Biology, National Research and Innovation Agency (BRIN), Cibinong, Bogor 16911, Indonesia
*Corresponding author’s email: sahirsabile@unhas.ac.id
Received: 12 September 2025 / Revised: 04 November 2025 / Accepted: 11 November 2025 / Published Online: 21 November 2025
Abstract
Semen quality is one of important factor that impacts cattle reproduction. However, the seminal plasma and spermatozoa separation is still unclear. We attempted proteomic and metabolomic techniques on fresh bovine semen and its compartments to molecularly characterise and cross these two compartments. The proteomic profile of the semen plasma showed pronounced accessory proteins enhancing lipid binding, ion homeostasis, and membrane dynamics, particularly PDC-109, enolases, VDAC2, and SP-10, while the spermatozoa comprised scaffolding, anchoring, and mitochondrial enzymatic proteins such as AKAP3, tektins, cylicins, and COX5B. The dichotomy was reinforced by complementary metabolomic analysis, with seminal plasma containing antioxidants and lipids such as taurine, ergothioneine, palmitoylglycine, and stearamide, while spermatozoa were enriched in metabolites associated with energy, including citrate, inosine, succinic semialdehyde, and pantothenic acid. Pathway analysis reinforced plasma specialisation about antioxidants and lipids rather than spermatozoa on glycolysis and amino acid metabolism with oxidative phosphorylation. Collectively, these results illustrate the presence of interconnected but non-interchangeable molecular domains in which seminal plasma provides protective and regulatory buffering, while spermatozoa are specialised in structural and energetic components for fertilisation. The candidate biomarkers identified from this study such as PDC-109, AKAP3, cylicins, taurine, and citrate illustrate molecular outputs corresponding to the quality of semen and provide a systematised context for enriched understanding of bovine reproductive biology.
Keywords: Bovine semen, Metabolomics, Seminal plasma, Spermatozoa, Proteomics
