Microbial diversity associated with the natural spring water of Western Himalayas

Springs in the Himalayan landscape are vital sources of potable water for millions of inhabitants who use it for domestic and farming purposes. The rise in urbanization, pilgrimage and various other anthropogenic activities invariably result in microbial contamination. In this study, we conducted a microbiological examination of water samples collected from four distinct Himalayan springs, located in the Rudraprayag district of Uttarakhand. We employed targeted metagenomics, using 16S rRNA and Internal Transcribed Spacer (ITS) gene analysis, to elucidate the bacterial and fungal composition. Cumulatively, the high-throughput sequencing data from all four springs revealed the abundance of Proteobacteria (35.38%), followed by Fermicutes (22.47%), Acidobacteriota (14.73%), and Actinobacteriota (5.35%). Genus level analysis of bacterial communities revealed the prevalence of Tumebacillus (13.85%) and Massilia (12.97%). Mycobiome analysis demonstrated the dominance of the fungal phyla Ascomycota (72.11%), followed by Basidiomycota (16.04%), Mortierellomycota (9.21%), and Chytridiomycota (1.2%). We further analysed the microbial communities using marker gene (16S rRNA) data through PICRUSt and the KEGG database to identify the gene abundance associated with carbon metabolism.