This study was conducted to investigate the effect of carotenoid, oligosaccharide and anthocyanin supplementation in broiler diets under
Escherichia coli lipopolysaccharide (LPS) challenge. Ross 308 chickens were fed 5 diets: basal diet (control diet), diet supplemented with β-glucan in 0.05% (positive control) and diets with 0.5% carotenoid-, oligosaccharide- or anthocyanin contents. On the 26th days of age, chickens were challenged intraperitoneally 2 mg LPS per kg of body weight. 12 h after injection, birds were euthanized, then spleen and ileum samples were collected. LPS induced increased relative mRNA expression of splenic (
p = 0.0445) and ileal (
p = 0.0435) interleukin-1β (
IL-1β), which was lower in the spleen in carotenoid (
p = 0.0114), oligosaccharide (
p = 0.0497) and anthocyanin (
p = 0.0303)-treated chickens compared to LPS-injected control birds. Dietary supplementation of carotenoids also decreased relative gene expression of splenic interleukin-6 (
IL-6) (
p = 0.0325). In the ileum, β-glucan supplementation showed lower relative mRNA expression of toll-like receptor 5 (
TLR-5) (
p = 0.0387) compared to anthocyanin treatment. Gene expression of both splenic and ileal interferon-α (
IFN-α), interferon-γ (
IFN-γ), toll-like receptor 4 (
TLR-4) and toll-like receptor 5 (
TLR-5) were not influenced by dietary supplements. In conclusion, carotenoids, oligosaccharides and anthocyanins could partially mitigate the immune stress caused by LPS challenge. All of the compounds impacted longer villus height (
p < 0.0001), villus height:crypt depth ratios were higher after β-glucan (
p < 0.0001) and anthocyanin (
p = 0.0063) supplementations and thickened mucosa was observed in β-glucan (
p < 0.0001), oligosaccharide (
p < 0.0001) and anthocyanin (
p = 0.048) treatments. All of these findings could represent a more effective absorption of nutrients.
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