Wastewater-Based Genomic Surveillance Reveals High-Risk Multidrug-Resistant Escherichia Coli And Klebsiella Pneumoniae In Windhoek, Namibia

Abstract

Wastewater-based epidemiology (WBE) provides a cost-effective approach to monitoring AMR dynamics in communities. This study used whole-genome sequencing (WGS) to investigate the genomic characteristics of multidrug-resistant Escherichia coli and Klebsiella pneumoniae isolated from four wastewater treatment plants (WWTPs) in Windhoek, Namibia. Influent and effluent samples (n =32) were collected and analyzed for physicochemical and microbiological parameters. Bacterial isolates were identified using Vitek and MALDI-TOF, followed by antimicrobial suscepti bility testing and WGS. Antibiotic resistance genes, plasmid replicons, virulence factors, and sequence types (STs) associated with each isolate were determined. High-risk clones, including E. coli ST410 and ST44 and K. pneumoniae ST219 and ST20, were detected within the treatment systems. Additional clinically relevant STs (e. g., ST10, ST10955, ST62) exhibited multidrug resistance profiles. Plasmid-borne ARGs, such as blaCTX-M-15, sul1, and tet(A/B), were detected in influent and effluent samples. Virulence determinants, including siderophore- mediated iron acquisition systems (yersiniabactin and enterobactin) and adhesion-associated fimbrial genes (fim and pap), were frequently detected alongside ARGs. Except for pH, temperature, and nitrate, most physico chemical parameters exceeded Namibian wastewater standards, indicating opportunities for further optimization of treatment processes. These findings highlight the value of wastewater-based genomic surveillance for char acterizing antimicrobial resistance in environmental settings and support its use to monitor the environmental distribution of multidrug-resistant bacteria in low- and middle-income countries (LMICs), particularly in settings where wastewater reuse is practiced.