Fig. 5: Changes in the diversity and composition of the sewage microbiome across different months (From June to December 2023) in Haryana.

a A 3D PCoA (Principal Coordinate Analysis) plot was generated to visualize variations in the sewage microbiome structure across different months. Each dot corresponds to a specific sample, with colors representing the respective months. The statistical significance of variance was assessed using permutational multivariate analysis of variance (PERMANOVA), revealing a significant variance among the sites (R2 = 0.213, p = 0.001). Box plots illustrating different alpha-diversity metrics include b Observed, c Shannon, and d Simpson indices, with statistical significance observed. Group differences were evaluated using the Kruskal-Wallis test, followed by pairwise comparisons with Dunn’s post hoc test. The Benjamini-Hochberg (BH) procedure was applied to adjust for multiple testing. The box represents the interquartile range (IQR), the median value is indicated by a line within the box, and whiskers extend to the extreme values (within 1.5 × IQR). The bars represent mean ± standard deviation, SD. e A heatmap depicting the distribution of highly prevalent bacterial taxa in terms of relative abundance at the species level across different months (from June to December 2023). The color code corresponds to the relative abundance of the bacterial taxa. The analysis included sewage samples collected longitudinally from June to December at multiple time points. The number of samples collected per month was as follows: June (n = 15), July (n = 28), August (n = 28), September (n = 28), October (n = 34), November (n = 18) and December (n = 21). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.