Tracking Age-Linked Antibiotic Resistance Patterns through Building-Level Wastewater Analysis

Antimicrobial resistance (AMR) is a global health challenge, and monitoring different demographic populations can improve our understanding of its spread and prevalence in urban settlements. This study applies building-level wastewater-based epidemiology (WBE) to analyze the resistome and mobilome of age-segregated populations from an elementary school (School), a university residence (UnivRes), and an elderly care facility (ElderlyRes) all located in Girona (Catalonia, Spain). Metagenomic analyses were subsequently conducted to investigate differences in bacterial communities, antibiotic resistance genes (ARGs), and mobile genetic elements (MGEs). The results revealed age-linked variations in the relative abundance and diversity of ARGs. The wastewater collected at the School exhibited the highest abundance of ARGs, while the ElderlyRes showed the highest diversity. Furthermore, sequences affiliated with bacterial pathogens were more prevalent in samples from both the School and the ElderlyRes, emphasizing potential public health implications. Among the 12 bacterial genera most strongly correlated with ARGs (Pearson <i>R</i> > 0.7), 11 were identified as members of the gut microbiota, underscoring their predominant role as reservoirs of resistance compared to bacteria of environmental origin. By integrating localized wastewater sampling with metagenomics, our study uncovers demographic-specific resistome patterns, delivering actionable evidence to strengthen AMR surveillance and intervention strategies in urban populations.