A putative model of the gut-muscle axis in aged livestock
The gut-muscle axis has been proposed to link gut microbiota with skeletal muscle physiology, yet its universality across livestock species remains unclear. Using aged laying hens, a livestock model with a relatively short digestive tract, we examined the gut microbiota, faecal metabolome, and breast-muscle metabolome by integrative multi-omics analyses in hens fed a Caldifermentibacillus hisashii-containing fermented feed or a control diet. Non-metric multidimensional scaling revealed clear separation of the microbial community between groups (stress = 0.0097), characterised by a marked expansion of Lactobacillus with the administration of the fermented feed. Variance partitioning showed that the 16S microbiota shared substantial variance with both the faecal (shared R2 adj = 0.54) and muscle (shared R2 adj = 0.48) metabolomes, and partial dbRDA demonstrated that the faecal-to-muscle metabolite association was largely retained after controlling for 16S (direct R2 = 0.538, partial R2 = 0.485), consistent with faecal metabolites acting as an integral layer linking microbiota to muscle. Cliff's delta-based selection showed depletion of proteolytic taxa and faecal amino acids, and reduced muscle Ornithine and uric acid alongside elevated Hypoxanthine. Because both groups were processed identically post-slaughter, these differences reflect in vivo states: amino acid depletion despite reduced bacterial proteolytic capacity points to enhanced host utilisation, and reduced uric acid, a post-mortem-stable purine end-product in uricotelic chickens, indicates efficient nitrogen turnover rather than accumulation. Collectively, these findings support a putative tripartite model of the gut-muscle axis in aged laying hens, providing a statistically grounded framework for understanding microbial contributions to muscle physiology in aged livestock.