Metagenomic insights into the urban–rural variation of antimicrobial resistance and pathogen reservoirs in untreated wastewater from central India

Introduction Rapid and scalable surveillance of antimicrobial resistance (AMR) is urgently needed in resource-constrained countries where routine monitoring is limited. Wastewater-based metagenomics offers a potential solution for early detection and geographic mapping of AMR. Methods We conducted a retrospective DNA shotgun metagenomic analysis of untreated wastewater collected across Nagpur, India (February–April 2021). A total of 422 grab samples were pooled into 138 composite samples from 10 urban zones and rural catchments. The bacterial microbiota and resistome were profiled, and urban–rural patterns were compared using diversity metrics and correlation analyses. Results Across all samples, 871 bacterial genera were detected, dominated by Proteobacteria, with frequent presence of Pseudomonas , Acinetobacter , Aeromonas , Acidovorax and Bacteroides . Beta diversity revealed statistically significant but subtle urban–rural compositional shifts. Of 33 globally important pathogens examined, 13 were detected at generally low relative abundance (