Cooked yolk fatty acid and metabolomic profiles reveal a greater docosahexaenoic acid proportion in one green-shell crossbred hen combination.

Although genetic background and crossbreeding affect egg quality and yolk composition, their effects on cooked-yolk lipid nutritional traits and metabolomic profiles remain unclear. This study evaluated production performance, internal and external egg quality , cooked-yolk fatty acids, and untargeted metabolomic profiles of four three-way cross combinations using G02, a Dongxiang green-shell chicken line, as the terminal sire at 30 weeks of age. The four combinations showed similar growth trajectories but differed in laying performance and selected egg-quality traits. Notably, G02 × (L02 × L03), involving Wenshang Barred (L02) and Tejia Barred (L03) maternal lines, exhibited lower total saturated, monounsaturated, and polyunsaturated fatty acid (PUFA) contents but higher eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) contents than the other combinations. It also showed the highest DHA/total PUFA proportion and omega-3/omega-6 fatty acid ratio. Untargeted metabolomics identified 11 candidate differentially abundant metabolites, mainly mapping to pathways related to carbon and energy metabolism. These findings indicate that G02 × (L02 × L03) showed a relatively favorable cooked-yolk long-chain n-3 fatty acid profile and may warrant further evaluation for developing green-shell eggs with a higher DHA proportion, although validation across laying stages and breeding batches is required.