Stimbiotics help improve intestinal immunity and positively modulate the gut microbiome in broilers with necrotic enteritis

Abstract

This experiment was conducted to investigate the effect of stimbiotic (STB) in broilers with necrotic enteritis (NE) on nutrient digestibility and gut health. A total of 200 one-day-old Arbor Acres (initial body weight of 44.03 ± 0.28 g) were used in this experiment for 28 days. All broilers were randomly allocated into four treatments, and each experimental group had 10 replicate cages with five broilers per cage. The experiment was conducted in a 2 × 2 factorial designs consisting of two levels of challenge (challenge and non-challenge) and two levels of STB (0 and 0.05 %). All broilers in challenged groups were orally challenged by overdosing with coccidia vaccines (× 10 recommended doses; Livacox® Q). The NE challenge significantly decreased (P < 0.05) nutrient digestibility, interferon-γ, heterophil levels in blood, and villus height:crypt depth (VH:CD) compared to the non-challenge group. Also, the NE challenge significantly lower (P < 0.05) ZO-1 and higher MUC2 gene expression than the non-challenge group. Supplementation of 0.05 % STB with NE challenge significantly increased (P < 0.05) gross energy digestibility and decreased (P < 0.05) the number of oocysts per gram of feces compared to the NE-challenged group. Supplementation of 0.05 % STB significantly increased (P < 0.05) the VH:CD in ileum compared to the non-supplementation group. Also, supplementation of 0.05 % STB is significantly lower (P < 0.05) MUC2 and TLR4 gene expression in ileum than the non-supplementation group. At the genus level, the supplementation of 0.05 % STB with NE challenge significantly decreased (P < 0.05) the abundance of Muribaculaceae compared to the NE-challenged group on d 21. In conclusion, supplementation of 0.05 % STB in a diet could positively regulate the cecal microflora and gene expression of tight junction protein and alleviate the decline in nutrient digestibility caused by NE.

Keywords: Broiler; Intestinal barrier integrity; Microbiome; Necrotic enteritis; Stimbiotic.

Fig. 1

Effects of supplementation with 0.05 % stimbiotic (STB) on the intestinal lesion score of broilers challenged with necrotic enteritis (NE) on day 21. -C, non-challenge; +C, NE challenge; 0, basal diet with non-supplementation of STB; 0.05, basal diet with supplementation of 0.05 % STB. n = 10 broilers/treatment. (A) x² = 42.794, P < 0.001. (B) x² = 21.333, P < 0.05. Numbers inside the bar indicate percentage of score out of total (100 %). a, b Means with different letters are significantly differ (P < 0.05).

Fig. 2

Effects of supplementation with 0.05 % stimbiotic (STB) on the intestinal lesion score of broilers challenged with necrotic enteritis (NE) on day 28. -C, non-challenge; +C, NE challenge; 0, basal diet with non-supplementation of STB; 0.05, basal diet with supplementation of 0.05 % STB. n = 10 broilers/treatment. (A) x² = 28.648, P < 0.001. (B) x² = 20.952, P < 0.05. Numbers inside the bar indicate percentage of score out of total (100 %). a, b Means with different letters are significantly differ (P < 0.05).

Fig. 3

Effects of supplementation with 0.05 % stimbiotic (STB) on the jejunum microscopic morphology (H&E staining) of broilers challenged with necrotic enteritis (NE) on day 21 and 28. -C, non-challenge; +C, NE challenge; 0, basal diet with non-supplementation of STB; 0.05, basal diet with supplementation of 0.05 % STB. The scale bar is 500 μm.

Fig. 4

Effects of supplementation with 0.05 % stimbiotic (STB) on the ileum microscopic morphology (H&E staining) of broilers challenged with necrotic enteritis (NE) on day 21 and 28. -C, non-challenge; +C, NE challenge; 0, basal diet with non-supplementation of STB; 0.05, basal diet with supplementation of 0.05 % STB. The scale bar is 500 μm.

Fig. 5

Unweighted and weighted Unifrac measurement in positive control (PC), basal diet; negative control (NC), basal diet with NE challenge; T1, PC + 0.05 % STB; T2, NC + 0.05 % STB. Each treatment group was placed as the control group, and treatment groups were compared by using one-way PROC MIXED with Dunnett’s post-hoc test.

Fig. 6

Visualized beta-diversity indices including unweighted and weighted emperor in positive control (PC), basal diet; negative control (NC), basal diet with NE challenge; T1, PC + 0.05 % STB; T2, NC + 0.05 % STB.