Metagenomic Analysis of Urban Wastewater Treatment Plant Effluents in Tokyo

Infect Drug Resist. 2022 Aug 23:15:4763-4777. doi: 10.2147/IDR.S370669. eCollection 2022.

Abstract

Purpose: Urban wastewater treatment plant (WWTP) effluents, even with proper treatment, may cause antimicrobial resistance (AMR) burden, with a high frequency of acquired antimicrobial resistance genes (ARGs). The dissemination of ARGs into the environment increases the risk of infectious diseases; however, there is little direct evidence regarding their epidemiological effects. This study aimed to assess effluents from urban WWTPs around the Tama River and Tokyo Bay using metagenomic analysis of (AMR) genes (ARGs) and heavy-metal resistance genes.

Methods: Metagenomic DNA-seq analysis of water samples and resistome analysis were performed.

Results: The most prevalent ARG was the sulfonamide resistance gene, sul1, followed by the quaternary ammonium compound resistance gene, qacE, suggesting that basic gene sets (sul1 and ∆qacE) in the class 1 integrons are the predominant ARGs. The aminoglycoside resistance genes, aadA and aph, and macrolide resistance genes, msr(E) and mph(E), were the predominant ARGs against each antimicrobial. bla OXA and bla GES were frequently detected, whereas the bla CTX-M cluster was faintly detected. Non-metric multidimensional scaling plot analysis and canonical correspondence analysis results suggested that marked differences in ARGs could be involved in the seasonal differences; qnrS2, aac(6')-Ib, and mef(C) increased markedly in summer, whereas msr(E) was more frequently detected in winter. Heavy-metal (Hg and Cu) resistance genes (HMRGs) were significantly detected in effluents from all WWTPs.

Conclusion: We characterized a baseline level of the environmental ARG/HMRG profile in the overall community, suggesting that environmental AMR surveillance, particularly in urban WWTPs, is a valuable first step in monitoring the AMR dissemination of bacteria from predominantly healthy individuals carrying notable ARG/Bs.

Keywords: ESBL; Enterobacteriaceae; carbapenemase; effluent; heavy-metal resistance; metagenomics; urban sewage.

Grants and funding

This work was supported by the Research Program on Emerging and Reemerging Infectious Diseases from the Japan Agency for Medical Research and Development (Grant numbers: JP19fk0108048 and JP20fk0108131). This work was also supported by a grant for research on emerging and reemerging infectious diseases and immunization (H30 Shinkogyosei-Ippan-002) from the Ministry of Health, Labour and Welfare, Japan. The funding agencies had no role in the study design, data collection and analysis, decision to publish, or manuscript preparation.