Anthropogenic activity remains the main contributor to fecal pollution in managed tropical watersheds as unraveled by PhyloChip microarray-based microbial source tracking

The spread of disease by enteric pathogens associated with fecal contamination is a major concern for the management of urban watersheds. So far, the relative contribution of natural and anthropogenic sources to fecal pollution in managed tropical watersheds remains poorly evaluated. In this study, the microbiomes of water samples collected from managed watersheds in Singapore were elicited using the PhyloChip, a dense 16S rRNA gene-based DNA microarray, and fecal impairment was inferred using a machine-learning classification algorithm (SourceTracker). The predicted contribution of wildlife fecal sources to environmental samples was generally negligible (< 0.01 +/- 0.01), indicating a low likelihood of fecal impairment from natural sources. However, sewage showed considerably higher contribution (0.09 +/- 0.05) to microbial communities in a subset of watershed samples from canals and rivers, suggesting persistent impairment of certain areas by anthropogenic activity although being managed. Interestingly, the contribution of sewage microbial communities showed decreasing trends from canals/rivers to the connected reservoirs, indicating meaningful auto-mitigation of fecal pollution in canals and rivers. Notably, exclusion of locally derived fecal samples and source categories from the training data set impaired the predictive performance of the classification algorithm despite a high degree of similarity in the phylogenetic composition of microbiomes in biologically similar but geographically distinct sources.