Whole-genome sequencing characterization of silver-resistant bacteria from the outfall of wastewater treatment plants and effluent-receiving rivers

ABSTRACT The excessive use of silver compounds has led to the environmental dissemination of silver resistance genes. However, little is known about the epidemiology of silver-resistant bacteria in the environment. Wastewater treatment plants (WWTPs) link the clinical settings with the natural environment and serve as a major pathway for silver entering the natural environment. However, their role in the dissemination of silver resistance genes remains unclear. This study investigated the characteristics of silver-resistant bacteria in the vicinity of four WWTPs to assess their environmental impact. Water and sediment samples were collected from the WWTP outfalls and downstream rivers. Among 22 silver-resistant strains obtained through plate screening, the majority were Klebsiella spp., followed by Escherichia spp. and Kluyvera spp. Notably, ST23 and ST2464 were the predominant sequence types (multilocus sequence typing) identified among the Klebsiella pneumoniae isolates. Antimicrobial susceptibility testing and whole-genome sequencing were performed to identify environmental heavy metal and antibiotic-resistant genes. Whole-genome sequencing revealed the presence of the sil and pco operons, which together formed the copper homeostasis and silver resistance island. The silver resistance gene sequences varied. Various heavy metal resistance genes, including mer and ars , were detected in the strains, as were a diverse array of plasmid types, including IncFIB(K) and repB(R1701), and fosA and β -lactamase encoding genes. Taken together, the findings underscore the coexistence of silver resistance genes with multiple heavy metal resistance genes in wastewater bacteria, highlighting the environmental implications of silver usage. Efforts should be directed toward restricting silver usage, improving WWTP purification methods to safeguard human and environmental health. IMPORTANCE The misuse of silver compounds has led to an increasing presence of silver-resistant microorganisms in the environment, which cannot be completely eliminated in wastewater treatment plants, allowing them to enter the environment and pose risks to environmental safety and human health. However, research on the epidemiology of silver-resistant bacteria in wastewater and their whole-genome sequencing remains limited. Our findings explain that silver-resistant bacteria from the environment often possess resistance to other heavy metals, share genetic similarities, and possess the potential for widespread transmission. Furthermore, these bacteria may enter clinical settings through environmental pathways, posing a risk to human health.