Modulation of Growth Performance and Intestinal Microbiota in Chickens Fed Plant Extracts or Virginiamycin

Abstract

In this study, the effects of plant extracts (PEs) and virginiamycin (VIRG) on broiler growth performance, as well as on host intestinal microbiota composition and function were investigated. A total of 288 one-day-old male Cobb broiler chickens were randomly divided into four treatment groups (with six replicates per group). The duodenal, ileal, and cecal content of six broilers per treatment group after 14 and 28 days of treatment were sampled. This material was used for high-throughput Illumina sequencing of the V3-V4 region of the 16S rRNA gene. The results showed that chickens fed 400 mg/kg plant extracts (HPE group) had significantly higher average body weights at day 28 as compared to the control group (CT; P < 0.05), and lower feed-to-meat ratios over days 15-42 (P < 0.01). Within the HPE group at day 14, the relative abundances of two bacterial phyla and 10 bacterial genera increased significantly in the ileal microbiota, and the relative abundance of three bacterial phyla and four bacterial genera decreased. The relative abundance of the genus Lactobacillus in the cecal microbiota decreased from 21.48% (CT group) to 8.41% (fed 200 mg/kg PEs; LPE group), 4.2% (HPE group), and 6.58% (fed 30 mg/kg virginiamycin; VIRG group) after 28 days. In contrast, Faecalibacterium and unclassified Rikenellaceae increased in abundance in the HPE group (from 18 to 28.46% and from 10.83 to 27.63%, respectively), while Bacteroides (36.7%) and Lachnospiraceae increased in abundance in the VIRG group. PICRUSt function analysis showed that the ileal microbiota of the PE treatment groups were more enriched in genes related to the meolism of cofactors and vitamins. In addition, the cecal microbiotas of the LPE and HPE groups were enriched in genes predicted to encode enzymes within 15 and 20 pathways, respectively. These pathways included protein digestion and absorption, amino acid metabolism, lipid biosynthesis, lipopolysaccharide biosynthesis, the citrate cycle (TCA cycle), and lipoic acid metabolism. Similarly, the VIRG group was enriched in 55 metabolic pathways (17 in the duodenum, 18 in the ileum, and 20 in the cecum) on day 28 (P < 0.05). Thus, the results indicated that the observed increase in broiler growth performance after PE or VIRG supplementation might be attributed to an improvement in intestinal microbial composition and metabolic function.

Keywords: broilers; growth performance; intestinal microbiota; plant extracts; virginiamycin.

FIGURE 1

Principal coordinate analysis (PLS-DA), based on the weighted unifrac distances of 16S rRNA bacterial sequences from the intestines of broilers. (A,B) PLS-DA of duodenal bacteria at (A) day 14 or (B) day 28. (C,D) PLS-DA of ileal bacteria at (C) day 14 or (D) day 28. (E,F) PLS-DA of cecal bacteria at (E) at day 14 or (F) day 28. CT, basal diet (control); VIRG, basal diet supplemented with 30 mg/kg virginiamycin; LPE, basal diet supplemented with 200 mg/kg plant extracts; HPE, basal diet supplemented with 400 mg/kg plant extracts.

FIGURE 2

Effects of plant extracts and virginiamycin on duodenal, ileal, and cecal microbiota composition at phylum and genus level. Relative abundances of bacterial phyla (A) and genera (B) in four treatment groups. CT, basal diet (control); VIRG, basal diet supplemented with 30 mg/kg virginiamycin; LPE, basal diet supplemented with 200 mg/kg plant extracts; HPE, basal diet supplemented with 400 mg/kg plant extracts. Intestinal chyme was collected from six birds per group for this analysis (n = 6).

FIGURE 3

Histograms showing the results of the linear discriminant analysis effect size (LEfSe) measurements, identifying the most differentially abundant phyla and genera between the PE/VIRG and CT groups. (A,B) Representation of the LEfSe results, showing different phyla and genera in the duodenum, ileum, and cecum samples from the HPE and CT chickens at day 14 and 28. (C,D) Representation of the LEfSe results, showing different phyla and genera in the duodenum, ileum, and cecum samples from the LPE and CT chickens at day 14 and 28. (E,F) Representation of the LEfSe results, showing different phyla and genera in the duodenum, ileum, and cecum samples from the VIRG and CT chickens at day 14 and 28. The CT chickens are indicated with a negative LDA scores (LDA > 2; P < 0.05). CT, basal diet (control); VIRG, basal diet supplemented with 30 mg/kg virginiamycin; LPE, basal diet supplemented with 200 mg/kg plant extracts; HPE, basal diet supplemented with 400 mg/kg plant extracts.

FIGURE 4

Genes overrepresented in the level 2 KEGG pathways in the duodenal, ileal, and cecal microbiotas of broilers on (A) day 14 and (B) day 28. CT, basal diet (control); VIRG, basal diet supplemented with 30 mg/kg virginiamycin; LPE, basal diet supplemented with 200 mg/kg plant extracts; HPE, basal diet supplemented with 400 mg/kg plant extracts.

FIGURE 5

Heatmap of relative abundance of predicted functional KEGG pathways of level 3 of the duodenal, ileal, and cecal microbiota of broilers on (A) day 14, and (B) day 28 by the LEfSe test (LDA > 2; P < 0.05). CT, basal diet (control); VIRG, basal diet supplemented with 30 mg/kg virginiamycin; LPE, basal diet supplemented with 200 mg/kg plant extracts; HPE, basal diet supplemented with 400 mg/kg plant extracts.

FIGURE 6

PICRUSt analysis of differences in metabolic functions among treatments, based on LEfSe results (LDA > 2; P < 0.05). (A,B) Differences in the metabolic functions of the intestinal microbiota between HPE and CT group chickens at (A) day 14 and (B) day 28. (C,D) Differences in the metabolic functions of the intestinal microbiota between LPE and CT group chickens at (C) day 14 and (D) day 28. (E) Differences in the metabolic functions of the intestinal microbiota between VIRG and CT group chickens at day 14. (F–H) Differences in the metabolic functions of the (F) duodenal, (G) ileal, and (H) cecal microbiota between VIRG and CT group chickens at day 28. CT, basal diet (control); VIRG, basal diet supplemented with 30 mg/kg virginiamycin; LPE, basal diet supplemented with 200 mg/kg plant extracts; HPE, basal diet supplemented with 400 mg/kg plant extracts.