Microbiota in Gut, Oral Cavity, and Mitral Valves Are Associated With Rheumatic Heart Disease

Figures

• 

  Figure 1. Alterations of gut microbiota in patients with rheumatic heart disease. Microbiota of fecal samples from patients with rheumatic heart disease (RHD) and control subjects (Ctrl) were analyzed using 16S rRNA gene sequencing. (A) Richness of fecal microbiota assessed by Chao1. (B) α-diversity of fecal microbiota assessed by Shannon index. (C) β-diversity analyzed by principal coordinate analysis (PCoA) based on Bray–Curtis distance of fecal microbiota at genus level. (D) Determination of module-based topological roles (peripherals, connectors, module hubs, or network hubs) of fecal microbiota in RHD patients using ZP-plot at genus level. The size of dots represents abundance of each genus. (E) Relative abundances of the fifteen most abundant genera in fecal microbiota. (F) Taxonomic cladogram of fecal microbiota based on LEfSe. Red color indicates increase and blue indicates decrease of taxa in RHD compared to Ctrl. LDA = 3. Mann–Whitney test was used. n = 20: 20. *P < 0.05; **P < 0.01; ****P < 0.0001.

• 

  Figure 2. Alterations of subgingival plaque microbiota in patients with rheumatic heart disease. Microbiota of subgingival plaques (SP) from patients with rheumatic heart disease (RHD) and control subjects (Ctrl) were analyzed using 16S rRNA gene sequencing. (A) Richness of SP microbiota assessed by Chao1. (B) α-diversity of SP microbiota assessed by Shannon index. (C) β-diversity analyzed by principal coordinate analysis (PCoA) based on Bray–Curtis distance of SP microbiota at genus level. (D) Determination of module-based topological roles (peripherals, connectors, module hubs, or network hubs) of SP microbiota in RHD patients using ZP-plot at genus level. The size of dots represents abundance of each genus. (E) Relative abundances of the fifteen most abundant genera in SP microbiota. (F) Taxonomic cladogram of SP microbiota based on LEfSe. Red color indicates increase and blue indicates decrease of taxa in RHD compared to Ctrl. LDA = 3. Mann–Whitney test was used. n = 20: 20. *P < 0.05.

• 

  Figure 3. Alterations of salivary microbiota in patients with rheumatic heart disease. Microbiota of saliva from patients with rheumatic heart disease (RHD) and control subjects (Ctrl) were analyzed using 16S rRNA gene sequencing. (A) Richness of salivary microbiota assessed by Chao1. (B) α-diversity of salivary microbiota assessed by Shannon index. (C) β-diversity analyzed by principal coordinate analysis (PCoA) based on Bray–Curtis distance of salivary microbiota at genus level. (D) Determination of module-based topological roles (peripherals, connectors, module hubs, or network hubs) of salivary microbiota in RHD patients using ZP-plot at genus level. The size of dots represents abundance of each genus. (E) Relative abundances of the fifteen most abundant genera in salivary microbiota. (F) Taxonomic cladogram of salivary microbiota based on LEfSe. Red color indicates increase and blue indicates decrease of taxa in RHD compared to Ctrl. LDA = 3. Mann–Whitney test was used. n = 20: 20. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

• 

  Figure 4. Comparisons of microbiota composition between mitral valves and other body sites of patients with rheumatic heart disease. Microbiota of mitral valves from patients with rheumatic heart disease (RHD) were analyzed using 16S rRNA gene sequencing. (A) Comparison of microbial β-diversity (PCoA) among gut, subgingival plaques (SP), saliva, and mitral valves (valve) of RHD patients. (B)Taxonomic composition of microbiota at different body sites of RHD patients at phylum level. (C) Illustration of genera detected in both fecal sample (f) and mitral valve (v) of each RHD patient. The numbers (1–16) denote RHD patients. The column on right indicates the number of patients who possess the corresponding genus in both sites. (D) Illustration of genera detected in both subgingival plaque (sp) and mitral valve (v) of each RHD patient. (E) Illustration of genera detected in both saliva (s) and mitral valve (v) of each RHD patient. n = 16.

• 

  Figure 5. Gut and oral microbiota differentiate rheumatic heart disease patients from control subjects. (A) Receiver operating characteristic (ROC) curve according to random forest model for fecal microbiota. A greater area under the ROC curve (AUC) indicates better performance. (B) ROC curve according to random forest model for subgingival plaque microbiota. (C) ROC curve according to random forest model for salivary microbiota.

• 

  Figure 6. Correlations between microbial genera and clinical indices in patients with rheumatic heart disease. (A) Heatmap showing correlations between different genera and LAD. (B) Heatmap showing correlations between different genera and PASP. LAD indicates left atrial diameter; and PASP, pulmonary artery systolic pressure. SP indicates subgingival plaque. Spearman’s rank correlation coefficients are illustrated in squares. *P < 0.05; **P < 0.01.