At 8 dpf, Lactobacillus group was significantly reduced in the TNBS-exposed groups (Figure 8B). Numbers of Burkholderia increased significantly (Figure 8D), but not Enterobacteriaceae family (Figure 8C). Which was consistent with the DGGE result. selleck screening library Figure 8 Quantitative

analysis of characteristic bacterial species. The relative quantity of specific groups of bacteria was determined by real-time PCR of 16S rRNA gene of (A) Total bacteria, (B) Lactobacillus group, (C) Burkholderia and (D) Enterobacteriaceae family. All reactions were performed in triplicate. Specific bacteria 16S rRNA gene amount was normalized to total bacteria 16S rRNA. Quantification values were represented as mean ± SEM log 16S rRNA gene copies per 10 ng of bacterial genomic DNA. a Indicates a significant difference (p<0.05) between TNBS-exposed group (25 μg/ml) and the control, b Indicates IWR-1 ic50 a significant difference (p<0.05) between TNBS-exposed group (50 μg/ml)

and the control, c Indicates a significant difference (p<0.05) between TNBS-exposed group (75 μg/ml) and the control, d Indicates a significant difference (p<0.05) between control groups at 6 dpf and 4 dpf, e Indicates a significant difference (p<0.05) between control groups at 8 dpf and 4 dpf. Enterocolitis severity and TNF-α expression correlate with the composition in gut microbiota We had observed the severity of enterocolitis in TNBS-treated zebrafish increased in a dose-dependent pattern at 8 dpf as compared with contols (Figure 2), whereas the abundance of Proteobacteria (especially Burkholderia) dramatically Stattic supplier increased and the proportion Interleukin-3 receptor of Firmicutes (Lactobacillus group) decreased significantly (Figure 8). We may predict that colitis severity would correlate with TNBS-induced changes in composition of gut bacteria. Accordingly, we calculated the correlation between enterocolitis scores and the density of Burkholderia

and Lactobacillus group separately by Pearson correlation analysis. We found that the colitis scores correlated with the abundance of Burkholderia (p=0.0045, Figure 9A) and the richness of Lactobacillus group (p=0.006, Figure 9B). These findings demonstrate that TNBS-induced enterocolitis correlate with changes in the composition and density of the gut microbiota. Figure 9 Total enterocolitis score and TNF-α expression level correlate with the composition of intestinal microbiota. A: Pearson correlation between total enterocolitis score and Lactobacillus group 16S rRNA gene copies, p=0.0045. B: Pearson correlation between total enterocolitis score and Burkholderia 16S rRNA gene copies, p=0.006. C: Pearson correlation between TNF-α expression levels and Lactobacillus group 16S rRNA gene copies, p=0.002. D: Pearson correlation between TNF-α expression levels and Burkholderia 16S rRNA gene copies, p=0.002.