A Comparative Study of Seminal Plasma and Sperm Metabolome Profiles in Toraya Buffalo Bulls

The objective of this study was to compare the metabolomic profiles of seminal plasma and sperm in Toraya buffalo bulls to identify key metabolites that influence sperm quality and reproductive potential. Semen samples were collected from eight mature Toraya buffalo bulls aged 4-10 years and classified as Saleko and Bonga types. Sperm were separated from seminal plasma and subjected to metabolite extraction, followed by derivatization and gas chromatography-mass spectrometry (GC-MS) analysis. Metabolites were identified and quantified using the NIST library, and statistical analysis was performed with MetaboAnalyst 6.0. This study utilized GC-MS to analyze the metabolomic profiles of seminal plasma and sperm from Toraya buffalo bulls. The analysis detected 10 metabolite groups, including carboxylic acids, fatty acids, and steroids. Metabolite enrichment revealed carboxylic acids as dominant in plasma and fatty acyls in sperm. Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) showed clear separation between seminal plasma and sperm. Key metabolites contributing to the separation were identified via variable importance in projection (VIP) analysis. Pathway enrichment analysis highlighted galactose metabolism in plasma and glyoxylate metabolism in sperm. Significant differences in metabolic pathways, such as citrate cycle and unsaturated fatty acid biosynthesis, were observed between the two samples. In conclusion, this study revealed unique metabolomic profiles in the seminal plasma and sperm of Toraya buffalo using GC-MS. Important metabolites such as creatinine, docosahexaenoic acid (DHA), and α-ketoglutaric acid may serve as potential biomarkers.