Natural intraepithelial lymphocyte populations rise during necrotic enteritis in chickens.

Intraepithelial lymphocytes (IEL) reside in the epithelium at the interface between the contents of the intestinal lumen and the sterile environment of the lamina propria. Because of this strategic location, IEL play a crucial role in various immunological processes, ranging from pathogen control to tissue stability. In mice and humans, IEL exhibit high diversity, categorized into induced IEL (conventional CD4 and CD8αß T cells) and natural IEL (TCRαßCD8αα, TCRγδ, and TCRneg IEL). In chickens, however, the subpopulations of IEL and their functions in enteric diseases remain unclear. Thus, we conducted this study to investigate the role of IEL populations during necrotic enteritis (NE) in chickens. At 14 days of age, sixty-three Specific-pathogen-free (SPF) birds were randomly assigned to three treatments: Control (sham challenge), Eimeria maxima challenge (EM), and Eimeria maxima + Clostridium Perfringens (C. Perfringens) co-challenge (EM/CP). The EM and EM/CP birds were infected with Eimeria maxima at day 14 of age, and EM/CP birds were additionally orally inoculated with C. perfringens at days 18 and 19 of age. Birds were weighed at days 18, 20, and 26 of age to assess body weight gain (BWG). At 20 days of age (1 day-post C. perfringens infection; dpi), and 26 days of age (7 dpi), 7 birds per treatment were euthanized, and jejunum was harvested for gross lesion scores, IEL isolation, and gene expression. The EM/CP birds exhibited subclinical NE disease, lower BWG and shorter colon length. The Most changes in the IEL populations were observed at 1 dpi. The EM/CP group showed substantial increases in the total number of natural IEL subsets, including TCRαß+CD4-CD8-, TCRαß+CD8αα+, TCRγδ+, TCRneg and innate CD8α (iCD8α) cells by at least two-fold. However, by 7 dpi, only the number of TCRαß+CD4-CD8- and TCRαß+CD8αα+ IEL maintained their increase in the EM/CP group. The EM/CP group had significantly higher expression of proinflammatory cytokines (IL-1ß and IFN-γ) and Osteopontin (OPN) in the jejunum at 1 dpi. These findings suggest that natural IEL with innate and innate-like functions might play a critical role in the host response during subclinical NE, potentially conferring protection against C. perfringens infection.

Effect of synbiotic supplementation on production performance and severity of necrotic enteritis in broilers during an experimental necrotic enteritis challenge.

To evaluate the efficacy of synbiotic during a necrotic enteritis (NE) infection, a total of 360 day-old chicks were randomly assigned into 4 experimental groups in a 2 × 2 factorial setup: control, challenge, synbiotic (1 g/kg), and challenge + synbiotic, with 6 replicates. NE was induced by gavaging 1 × 104Eimeria maxima oocysts and 1 × 108 CFU/mL of Clostridium perfringens on d 14 (D14) and D19, 20, and 21, respectively. At D35, the NE challenge decreased the BW gain (P < 0.001) and increased feed conversion ratio (P = 0.03), whereas synbiotic supplementation decreased the feed intake (P = 0.04). At D21, NE challenge increased gut permeability (P < 0.001), decreased regulatory T cells (Tregs) in the cecal tonsil (CT) (P = 0.02), increased Tregs in the spleen (P = 0.02), decreased nitric oxide (NO) production in the spleen (P = 0.04) and decreased IL-10 expression in CT (P = 0.02), whereas synbiotic supplementation increased CD4+:CD8+ T cells in the spleen (P < 0.001) and decreased interferon (IFN)-γ expression in the jejunum (P = 0.07), however, synbiotic supplementation during NE challenge decreased mid-gut lesion score (P < 0.001), increased CD4+:CD8+ T cells in CT and decreased IgA production in bile (P < 0.001), compared to the control group. At D28, synbiotic supplementation decreased CD4+:CD8+ T cells in CT (P < 0.001), whereas synbiotic supplementation during NE challenge decreased Tregs in CT (P < 0.001) and increased NO production in the spleen (P = 0.04), compared to the control group. At D35, the NE challenge decreased CD4+:CD8+ T cells in the spleen (P = 0.03), decreased IgA production in bile (P = 0.02), decreased IL-10 expression in CT (P = 0.04), and decreased IL-10 (P = 0.009), IFN-γ (P = 0.03) and inducible nitric oxide synthase (P = 0.02) expression in the jejunum, whereas synbiotic supplementation increased Tregs in the spleen (P = 0.04), compared to control group. Synbiotic supplementation during the NE challenge decreased both IL-1ß (P = 0.02) and IFN-γ (P = 0.001) expression in CT, compared to the control group. It can be concluded that synbiotic supplementation increases production performance by decreasing mid-gut lesions and enhancing protective immunity against NE, and efficiency of synbiotic could be improved by blending additional probiotics and prebiotics.