The Yoruba Ecotype Chicken (YEC) is an indigenous Nigerian poultry breed of considerable importance. The metabolomic composition of its sperm reserves remains uncharacterized, with limiting molecular-level understanding of its reproductive biology. The present study was aimed to quantify metabolites in sperm reserves of YEC, map them to annotated metabolic pathways, and integrate them with sperm quality parameters to derive candidate fertility biomarkers. Thirty sexually mature cocks of 25-30 weeks age were used. Semen was collected from these birds via abdominal massage method. Sperm reserves were analyzed using an Agilent 3800 Gas Chromatography with a 4000 MSD and HP-5 Mass Spectrophotometry column. Methyl chloroformate was used for pre-column derivatization. Metabolites were identified via the NIST08/Wiley275 library, mapped to Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, and integrated with sperm progressive motility, concentration, acrosomal competitive index (ACI), membrane evaluation index (MEI), and morphological abnormalities. Results showed that 16 metabolites were identified across eight biochemical classes. The metabolome was dominated by dl-2-Aminobutyric acid (17.24±12.05%), L-Glutamine (12.50±1.61%), and cholesterol (10.04±0.95%). Metabolites mapped to 11 KEGG-annotated pathways. Sperm quality was excellent, with progressive motility 77.50±1.44%, MEI 80.00±0.58%, ACI 71.75±7.26%, sperm viability 63.09±2.53% and morphological abnormalities 3.00±0.71%. In conclusion, YEC sperm reserve metabolome reflects active energy metabolism, antioxidant defense, and membrane maintenance. L-Glutamine, cholesterol, and dl-2-Aminobutyric acid are proposed as primary biomarkers of sperm reserve quality. This study delivers the first comprehensive GC–MS-based metabolomic profile of sperm reserves in YEC, integrating data with sperm quality parameters to propose a tier-ranked fertility biomarker panel. These findings support conservation and productivity of indigenous poultry, contributing to Zero Hunger through enhanced reproductive efficiency in smallholder systems.

To Cite This Article: Akintunde AO, Mustofa I, Ndubuisi-Ogbonna LC, Aliyu KI, Widodo OS, Khan IU, Maslachah L and Puspitasari Y, 2026. Metabolomic profiles of sperm reserves as potential fertility biomarkers for yoruba ecotype chickens. Pak Vet J, 46(8): 2108-2117. http://dx.doi.org/10.29261/pakvetj/2026.193