Dietary supplementation with berberine improves growth performance and modulates the composition and function of cecal microbiota in yellow-feathered broilers

Abstract

This study investigated the effect of berberine (BBR) on growth performance and composition and function of cecal microbiota in yellow-feathered broilers. A total of 360 1-day-old female broilers were assigned to 3 dietary treatments, each with 6 replicates of 20 birds. The dietary treatments consisted of a basal diet as negative control (NC), basal plus 200 mg/kg oxytetracycline calcium and 250 mg/kg nasiheptide as an antibiotic positive control (PC), and basal plus 250 mg/kg BBR. On day 21, 42, and 63, one chicken from each replicate was randomly selected for blood collection and cecal sampling. The 16S rRNA sequencing technology was used to analyze the community composition and function of cecal microbiota. Dietary supplementation with antibiotics or BBR increased the final body weight (BW) at day 63 and the average daily gain (ADG) during 1 to 21 d compared with the NC (P < 0.05). Supplementation with BBR improved the average daily feed intake (ADFI) at 22 to 42 d, 43 to 63 d, and 1 to 63 d (P < 0.05). Feed efficiency, indicated by feed to gain ratio (F/G), increased with PC during day 1 to 21 compared with NC (P < 0.05). The plasma concentrations of total protein at 42 d and uric acid at 21 d were increased, whereas creatine concentration at 63 d was decreased by BBR treatment (P < 0.05). The Chao 1 and Shannon index representing microbial α-diversity was reduced by BBR treatment (P < 0.05). The abundances of phylum Firmicutes and genera Lachnospiraceae, Lachnoclostridium, Clostridiales, and Intestinimonas were decreased, whereas the abundances of phylum Bacteroidetes and genus Bacteroides were increased with BBR treatment. Functional prediction of microbiota revealed that BBR treatment enriched pathways related to metabolism, organismal systems, and genetic information processing, especially DNA replication. The abundance of phylum Bacteroidetes, and genera Bacteroides and Lactobacillus in cecal contents were positively correlated with broiler growth performance. These results demonstrated dietary BBR supplementation improved the growth performance of yellow-feathered broilers, and was closely related to the significant changes in cecal microbiota composition.

Keywords: berberine; broiler; cecum; growth performance; gut microbiota.

Figure 1

Effect of dietary berberine supplementation on the composition of cecal microbiota in yellow-feathered broilers at the phylum and genus levels. The relative abundance of the most abundant 10 phyla in cecal contents of yellow-feathered broilers (A) and the significantly different species at the phylum level among treatments (B). The relative abundance of the most abundant 10 genera in cecal contents of yellow-feathered broilers (C) and the significantly different species at genus level among treatments (D). Differences were determined by one-way ANOVA followed by Duncan's multiple comparison. ^a,b indicate a significant difference at P < 0.05 (n = 6). Abbreviations: BBR, berberine supplementation at 250 mg/kg; NC, negative control without in-feed antibiotics; PC, positive control with in-feed antibiotics 200 mg/kg of oxytetracycline calcium and 250 mg/kg of nasiheptide.

Figure 2

Effect of dietary berberine supplementation on the α-diversity of cecal microbiota in yellow-feathered broilers. (A) Venn diagram of OTU. (B) Chao 1. (C) Shannon index. ∗indicates a significant difference at P < 0.05. Abbreviations: BBR, berberine supplementation at 250 mg/kg; NC, negative control without in-feed antibiotics; PC, positive control with in-feed antibiotics 200 mg/kg of oxytetracycline calcium and 250 mg/kg of nasiheptide.

Figure 3

Effect of dietary berberine supplementation on the β-diversity of cecal microbiota in yellow-feathered broilers. (A) PCoA plot. (B) Analysis of similarities (ANOSIM) tests were performed between groups based on relative abundance of OTU. (C) The LEfSe analysis (LDA score ≥ 4). (D) UPGMA Clustering was conducted based on Unweighted Unifrac distance. Abbreviations: BBR, berberine supplementation at 250 mg/kg; LEfSe, linear discriminant analysis effect size; NC, negative control without in-feed antibiotics; PC, positive control with in-feed antibiotics 200 mg/kg of oxytetracycline calcium and 250 mg/kg of nasiheptid; PCoA, Principal Coordinate Analysise; UPGMA, Unweighted Pair-group Method with Arithmetic Means.

Figure 4

T-test analysis for the significant changes of differential cecal microbiota at different levels in yellow-feathered broilers. (A) Comparison of differential cecal microbiota at day 42. (B) Comparison of differential cecal microbiota at day 63. Abbreviations: BBR, berberine supplementation at 250 mg/kg; NC, negative control without in-feed antibiotics; PC, positive control with in-feed antibiotics 200 mg/kg of oxytetracycline calcium and 250 mg/kg of nasiheptide.

Figure 5

Predicted microbial function of cecal microbiota in yellow-feathered broilers treated with berberine. The heatmap of predicted microbial function by PICRUST at day 21 (A), 42 (B), and 63 (C). The comparisons of cecal microbial functions of yellow-feathered broilers between treatments and ages (D–G). When compared to the negative controls (NC), BBR significantly increased the function enrichment of DNA replication, starch and sucrose metabolism, transcription machinery, chaperones and folding catalysts, and protein export in the cecal microbiota of 42-d yellow-feathered broilers. At day 63, BBR significantly increased the function enrichment of DNA replication, carbon fixation pathways in prokaryotes, glycine, serine and threonine metabolism, and protein export when compared with controls (NC). Statistics were conducted by two-sided Welch's t-test and Benjamini-Hochberg FDR correction between pairs of means, and the P-value of different functions lower than 0.05 are shown. Abbreviations: BBR, berberine supplementation at 250 mg/kg; NC, negative control without in-feed antibiotics; PC, positive control with in-feed antibiotics 200 mg/kg of oxytetracycline calcium and 250 mg/kg of nasiheptide.

Figure 6

The Spearman correlation analyses of cecal microbial species with growth performance in yellow-feathered broilers. Relationships between growth performance and cecal microbial species at the phylum (A–C) or genus level (D–F) determined at day 21, 42, or 63. Performance during the starter phase (A, B), grower phase (C, D), finisher phase and overall (E, F) are shown as heatmaps with red and blue representing positive correlation and negative correlations. ADGW, ADFIW, and FGW, are the ADG, ADFI, and F/G during the whole period (1 to 63 d). ∗indicates a significant difference at P < 0.05, and ∗∗ indicates P < 0.01. Abbreviation: FG, feed-to-gain ratio.