Interphylum dissemination of NDM-5-positive plasmids in hospital wastewater from Fuzhou, China: a single-centre, culture-independent, plasmid transmission study

作者：Qiu E. Yang, Xiaodan Ma, Lingshuang Zeng, Qinqin Wang, Minchun Li, Lin Teng, Mingzhen He, Chen Liu, Mengshi Zhao, Mengzhu Wang, Hui Deng, Jonas Stenløkke Madsen, Hanpeng Liao, Timothy R. Walsh, Shungui Zhou · 发表于：The Lancet Microbe · 年份：2023 · DOI：10.1016/s2666-5247(23)00227-6 · 被引用次数：69 · 研究领域：Antibiotic Resistance in Bacteria、Pharmaceutical and Antibiotic Environmental Impacts、Antibiotic Use and Resistance

BACKGROUND: The global spread of plasmid-borne carbapenem resistance is an ongoing public health challenge; however, the nature of such horizontal gene transfer events among complex bacterial communities remains poorly understood. We examined the in-situ transfer of the globally dominant New Delhi metallo-β-lactamase (NDM)-5-positive IncX3 plasmid (denoted pX3_NDM-5) in hospital wastewater to simulate a real-world, One Health antimicrobial resistance context. METHODS: For this transmission study, we tagged pX3_NDM-5 with the green fluorescent protein gene, gfp, using a CRISPR-based method and transferred the plasmid to a donor Escherichia coli strain. Bacteria were extracted from a hospital wastewater treatment plant (Fujian Provincial Maternity and Children's Hospital, Fuzhou, China) as the bacterial recipient community. We mixed this recipient community with the E coli donor strain carrying the gfp-tagged plasmid, both with and without sodium hypochlorite (NaClO) as an environmental stressor, and conducted several culture-based and culture-independent conjugation assays. The conjugation events were observed microscopically and quantified by fluorescence-activated cell sorting. We analysed the taxonomic composition of the sorted transconjugal pool by 16S rRNA gene amplicon sequencing and assessed the stability of the plasmid in the isolated transconjugants and its ability to transfer back to E coli. FINDINGS: We show that the plasmid pX3_NDM-5 has a broad host range and can ...