Specific Secondary Bile Acids Control Chicken Necrotic Enteritis - PubMed (original) (raw)

Specific Secondary Bile Acids Control Chicken Necrotic Enteritis

Mohit Bansal et al. Pathogens. 2021.

Abstract

Necrotic enteritis (NE), mainly induced by the pathogens of Clostridium perfringens and coccidia, causes huge economic losses with limited intervention options in the poultry industry. This study investigated the role of specific bile acids on NE development. Day-old broiler chicks were assigned to six groups: noninfected, NE, and NE with four bile diets of 0.32% chicken bile, 0.15% commercial ox bile, 0.15% lithocholic acid (LCA), or 0.15% deoxycholic acid (DCA). The birds were infected with Eimeria maxima at day 18 and C. perfringens at day 23 and 24. The infected birds developed clinical NE signs. The NE birds suffered severe ileitis with villus blunting, crypt hyperplasia, epithelial line disintegration, and massive immune cell infiltration, while DCA and LCA prevented the ileitis histopathology. NE induced severe body weight gain (BWG) loss, while only DCA prevented NE-induced BWG loss. Notably, DCA reduced the NE-induced inflammatory response and the colonization and invasion of C. perfringens compared to NE birds. Consistently, NE reduced the total bile acids in the ileal digesta, while dietary DCA and commercial bile restored it. Together, this study showed that DCA and LCA reduced NE histopathology, suggesting that secondary bile acids, but not total bile acid levels, play an essential role in controlling the enteritis.

Keywords: Clostridium perfringens; Eimeria maxima; intestinal inflammation; microbiota; secondary bile acid.

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Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Figures

Figure 1

Figure 1

DCA Prevented Clinical NE-induced Ileal Inflammation. Cohorts of 7 to 14 broiler chickens were fed basal diets supplemented with different bile acids starting from day 14. Birds were challenged with E. maxima at day 18 and C. perfringens at day 23 and 24. The birds were sacrificed on day 25. (A) Representative images of H&E staining showing small intestinal histopathology. (B) Microscopic quantification of intestinal histopathological lesion score. Yellow arrows: immune cell infiltration; Green arrow: blunted villi; blue arrow: hyperplastic crypts. Different letters of a, b, and c mean p < 0.05. The scale bar is 200 μm. Results are representative of 3 independent experiments.

Figure 2

Figure 2

Cohorts of broiler chickens were fed different bile diets and infected as in Figure 1. (A) Representative images of the TUNEL assay showing cell death at the late stage of apoptosis (green dots). Yellow arrowhead: apoptotic immune cells in lamina propria; Green arrow: apoptotic epithelial cells. (B) Ileal Ifnγ and the Mmp9 mRNA qPCR fold-change relative to noninfected birds and normalized to Gapdh. Different letters of a and b mean p < 0.05. The scale bar is 40 μm. Results are representative of 3 independent experiments.

Figure 3

Figure 3

DCA reduced C. perfringens and E. maxima ileal colonization. Cohorts of broiler chickens were fed different bile diets and infected as in Figure 1. (A) Luminal C. perfringens colonization was quantified in the ileal digesta by measuring 16S rDNA using a qPCR. (B) Luminal E. maxima was quantified in the ileal digesta by measuring 18S rDNA using a qPCR. (C) C. perfringens mucosal invasion was visualized by FISH. Yellow arrows indicate the vegetative and spore cells (red) of C. perfringens. Yellow arrowheads indicate red blood cells (red). The blue color is the DAPI staining for the cell nucleus. The scale bar is 20 µm. All graphs show mean ± SEM. Different letters of a, b, and c mean p < 0.05. Results are representative of 3 independent experiments.

Figure 4

Figure 4

DCA reduced NE-induced productivity loss. Cohorts of broiler chickens were fed different bile supplemented diets from day 14 and infected as in Figure 1. Bird body weight gain (BWG) was measured at 0, 14, 18, and 25 days of age, and daily periodic BWG is shown. All graphs show mean ± SEM. Different letters of a, b, and c mean p < 0.05. Results are representative of 3 independent experiments.

Figure 5

Figure 5

Dietary bile acids modulated the ileal bile acid composition in NE birds. Cohorts of broiler chickens fed with different bile acids and infected with NE as in Figure 1. (A) Total and individual bile acid quantification. (B) Relative composition of bile acids. All graphs show mean ± SEM. Different letters of a, b, and c mean p < 0.05. Results are representative of 3 independent experiments.

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