Gallstone and bile microbiota: A case-control study based on 16S rRNA gene sequencing

Figure 1 Clinical characteristics of the study subjects. ^aP < 0.05 vs control was considered statistically significant. Differences in clinical characteristics between the groups were analyzed using the Student’s t-test. A: Abdominal B-ultrasound in patients with gallstones, purple boxes point to gallstones; B: Computed tomography mage in patients with gallstones, and the high-density highlight in the purple box is the gallstone; C: Comparison of serum biochemical parameters including total bile acids, triglycerides, cholesterol, high-density lipoprotein, glucose, serum total bilirubin, and low-density lipoprotein between the two groups. TBA: Total bile acids; TG: Triglycerides; CH: Cholesterol; HDL: High-density lipoprotein; GLU: Glucose; STB: Serum total bilirubin; LDL: Low-density lipoprotein; GS: Gallstone; GSF: Gallstone-free.

Figure 2 Analysis of 16S rRNA gene sequences and bile microbiome diversity. Statistical significance vs the control was defined as a two-tailed ^aP < 0.05. A: Venn on operational taxonomic units level; B: Rank-Abundance curves on operational taxonomic units level; C: Alpha diversity analysis of bile microbiome including Shannon, Chao, Simpson, Ace, Coverage, and Sobs index; D: Beta diversity analysis of bile microbiome. OUT: Operational taxonomic units; GS: Gallstone; GSF: Gallstone-free; PCoA: Principal coordinate analysis; PC1: Principal component 1; PC2: Principal component 2.

Figure 3 Analysis of the composition of bile microorganisms. A: Community heatmap analysis at genus level, the red part indicates that the abundance of the genus in this sample is higher than other samples; the blue part represents the lower abundance of the genus in this sample than in other samples; B: Community barplot analysis at Phylum and Genus level. GS: Gallstone; GSF: Gallstone-free.

Figure 4 Analysis of the differences of bile microorganisms. A: Analysis of bile microorganisms at the genus level between the two groups was performed using Student’s t-test, with a P value < 0.05 vs control indicating significant differences in bacterial genera; B: Linear discriminant analysis effect size multi-level species differential analysis was conducted in the two groups, significance thresholds were set at Linear Discriminant Analysis Effect Size Score (log10) > 3.0 and P < 0.05 vs control. GS: Gallstone; GSF: Gallstone-free.