Global overview of sputum microbiome. (a) A stacked bar chart to show the top 20 most represented microbiome genera in TB (SP12-SP61) and non-TB (SP313-SP321) sputa. SP47 had an expansion of Haemophilus and SP315 comprised mainly of known artifacts Ralstonia and Bradyrhizobium. These two samples were subsequently removed from all downstream analyses. (b) Microbiome species richness and diversity were calculated. Non-TB samples (n = 9) had a significantly higher number of observed OTUs and estimated number of true OTUs (chao1 indicator) compared to TB samples (n = 17). (c) There was no difference in species diversity, as measured by the Shannon and Simpson indices, indicating species evenness and distribution did not differ between TB and non-TB groups. (d) HIV-1 coinfection did not impact the global microbiome species richness or diversity in sputum. For panels b to d, statistical difference was calculated using Mann-Whitney U-test (*, P < 0.05; **, P < 0.01; n.s., not significant).