Porcine Bile Acids Improve Antioxidant Status and Immune Function by Increasing Hungatella Abundance with Different Protein Level Diets in Late-Laying Hens

The modern livestock industry faces significant challenges due to the extension of production cycles and a growing shortage of feed resources. Low-protein (LP) diets offer potential environmental advantages whereas damage intestinal integrity and increase pro-inflammatory cytokines. Bile acids (BAs), metabolic products of cholesterol in the liver, possess strong antioxidative and immune properties. This study evaluated the effects of dietary porcine BA supplementation at different protein levels on the antioxidant status, immune function, and gut microbiota of late-laying hens. A 2 × 2 factorial design was employed with 198 Hy-Line Brown laying hens (62 weeks old), randomly divided into one of four treatment groups with 8 replicates for each treatment. Diets included normal (16.42%) or low-protein (15.35%) levels, with porcine BAs supplemented at 0 or 120 mg/kg (62-69 weeks) and 0 or 200 mg/kg (70-75 weeks) over 14 weeks. Dietary protein levels and bile acids had no effect on organ development in laying hens. Low-protein diets increased serum anti-inflammatory cytokines IL-4 and IL-10 but elevated pro-inflammatory cytokines IL-1β (<i>p</i> < 0.05), corroborated by higher <i>IL-10</i> gene expression in the ileum mucosa (<i>p</i> < 0.05). The supplementation of bile acids improved immune function by increasing serum IL-4 and TGF-β while decreasing IL-6 levels (<i>p</i> < 0.05). Moreover, bile acids treatments upregulated <i>IL-10</i>, <i>TGF-β</i> and <i>ZO-1</i> gene expression in the ileum mucosa (<i>p</i> < 0.05) and alleviated oxidative stress by enhancing serum GSH-Px activity (<i>p</i> < 0.05). Additionally, both low-protein diets and bile acids enriched beneficial cecal bacteria, including <i>Hungatella</i> and <i>Blautia</i>, primarily linked to immune function. These findings demonstrate that porcine bile acids administration improve antioxidant capacity, immune function and gut microbiota dysbiosis. These beneficial effects were associated with <i>Hungatella</i> and <i>Blautia</i> levels, which may be responsible for the porcine bile acids efficacy in poultry nutrition.