The Glutamatergic System Regulates Feather Pecking Behaviors in Laying Hens Through the Gut-Brain Axis

Feather pecking (FP) is a significant welfare and economic problem in laying hen husbandry. While there is growing evidence that the glutamatergic system plays a crucial role in regulating FP behavior, the biological mechanisms remain unclear, largely due to the limited uptake of peripheral glutamate across the blood-brain barrier (BBB). Here, we applied a multi-omics approach combined with physiology assays to answer this question from the perspective of the gut-brain axis. A total of 108 hens were randomly assigned to two groups (treatment and control) with six replicates each, and the treatment group was subjected to chronic environmental stressors including re-housing, noise, and transport. We found that chronic exposure to environmental stressors induced severe FP, accompanied by reduced production performance and increased anxiety- and depression-related behaviors, compared to controls. In addition, the immune system was potentially disrupted in FP chickens. Notably, gut microbiota diversity and composition were significantly altered, leading to decreased microbial community stability. Non-targeted metabolomic analysis identified a variety of differential metabolites, primarily associated with arginine and histidine biosynthesis. A significant increase in glutamate levels was also observed in the hippocampus of FP chickens. Transcriptome analysis revealed the upregulated expressions of glutamate-related receptors GRIN2A and SLC17A6 in the hippocampus. Correlation analysis indicated that GRIN2A and SLC17A6 are positively associated with arginine levels in the duodenum, while Romboutsia in the duodenum is negatively correlated with arginine. These findings suggest that intestinal bacteria, including Romboutsia, may influence FP behavior by altering plasma arginine and histidine levels. These changes, in turn, affect glutamate levels and receptor gene expression in the hippocampus, thereby regulating the glutamatergic system. Our research offers insights into novel strategies for mitigating harmful behaviors in poultry farming, with potential benefits for animal performance and welfare.