Effect of Carotenoids, Oligosaccharides and Anthocyanins on Growth Performance, Immunological Parameters and Intestinal Morphology in Broiler Chickens Challenged with Escherichia coli Lipopolysaccharide

Abstract

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.

Keywords: anthocyanins; broiler chicken; carotenoids; cytokines; gene expression; intestinal morphology; natural compounds; oligosaccharides; receptors; β-glucan.

Figure 1

HPLC profile of DAD detection of carotenoids on 460 nm from Hungarian red sweet pepper.

Figure 2

Relative spleen weight (spleen weight compared to live weight) of chickens fed basal diet under Escherichia coli O55:B5 LPS challenge, basal diet under isotonic saline challenge, diet supplemented with 0.05% β-glucan-, diet supplemented with 0.5 % carotenoids-, diet supplemented with 0.5% oligosaccharides- and diet supplemented with 0.5% anthocyanins under Escherichia coli O55:B5 LPS challenge (n = 6/treatment). Error bars represent means ± standard errors of the mean. The effects were analyzed by One-way ANOVA and differences among treatments were considered significant at p < 0.05. Dietary effects were not significant.

Figure 3

Relative interleukin-1β (a), interleukin-6 (b), interferon-α (c), interferon-γ (d), toll-like receptor 4 (e) and toll-like receptor 5 (f) mRNA expression in spleen of chickens fed basal diet under Escherichia coli LPS challenge, basal diet under isotonic saline challenge, diet supplemented with 0.05% β-glucan-, diet supplemented with 0.5% carotenoids-, diet supplemented with 0.5% oligosaccharides- and diet supplemented with 0.5% anthocyanins under Escherichia coli O55:B5 LPS challenge (n = 6/treatment). Error bars represent means ± standard errors of the mean. The effects were analyzed by One-way ANOVA and groups that do not share a letter are significantly different (p < 0.05).

Figure 3

Relative interleukin-1β (a), interleukin-6 (b), interferon-α (c), interferon-γ (d), toll-like receptor 4 (e) and toll-like receptor 5 (f) mRNA expression in spleen of chickens fed basal diet under Escherichia coli LPS challenge, basal diet under isotonic saline challenge, diet supplemented with 0.05% β-glucan-, diet supplemented with 0.5% carotenoids-, diet supplemented with 0.5% oligosaccharides- and diet supplemented with 0.5% anthocyanins under Escherichia coli O55:B5 LPS challenge (n = 6/treatment). Error bars represent means ± standard errors of the mean. The effects were analyzed by One-way ANOVA and groups that do not share a letter are significantly different (p < 0.05).

Figure 4

Relative interleukin-1β (a), interferon-α (b), interferon-γ (c), toll-like receptor 4 (d) and toll-like receptor 5 (e) mRNA expression in ileum of chickens fed basal diet under Escherichia coli LPS challenge, basal diet under isotonic saline challenge, diet supplemented with 0.05% β-glucan-, diet supplemented with 0.5% carotenoids-, diet supplemented with 0.5% oligosaccharides- and diet supplemented with 0.5% anthocyanins under Escherichia coli O55:B5 LPS challenge (n = 6/treatment). Error bars represent means ± standard errors of the mean. The effects were analyzed by One-way ANOVA and groups that do not share a letter are significantly different (p < 0.05).

Figure 4

Relative interleukin-1β (a), interferon-α (b), interferon-γ (c), toll-like receptor 4 (d) and toll-like receptor 5 (e) mRNA expression in ileum of chickens fed basal diet under Escherichia coli LPS challenge, basal diet under isotonic saline challenge, diet supplemented with 0.05% β-glucan-, diet supplemented with 0.5% carotenoids-, diet supplemented with 0.5% oligosaccharides- and diet supplemented with 0.5% anthocyanins under Escherichia coli O55:B5 LPS challenge (n = 6/treatment). Error bars represent means ± standard errors of the mean. The effects were analyzed by One-way ANOVA and groups that do not share a letter are significantly different (p < 0.05).