上海市饮用水源地原水中抗生素抗性基因多样性研究  

Diversity of antibiotic resistance genes in raw water from drinking water source in Shanghai

作　　者：潘淼 朱彦祺 方沚昕 张曦[1] 陈洪友[1] 陈敏[1] PAN Miao;ZHU Yanqi;FANG Zhixin;ZHANG Xi;CHEN Hongyou;CHEN Min(Division of Pathogen Testing and Analysis,Shanghai Municipal Center for Disease Control and Prevention,Shanghai 200336,China)

机构地区：[1]上海市疾病预防控制中心病原生物检定所,上海200336

出　　处：《上海预防医学》2022年第11期1123-1128,共6页Shanghai Journal of Preventive Medicine

基　　金：上海市卫生健康委科研课题(20184Y0292);上海市公共卫生体系建设三年行动计划人才培养计划(GWV-10.2-YQ04);上海市公共卫生体系建设三年行动计划重点学科项目(GWV-10.1-XK3)。

摘　　要：【目的】研究饮用水源地原水中的各种抗生素抗性基因(ARGs)的分布,分析抗性基因与水源地水中分离到的常见碳青霉烯多重耐药细菌相关性,为城市饮用水安全提供科学依据。【方法】采集2020年上海市某主要饮用水源地原水水样30件,用含1μg·μL^(−1)美罗培南的哥伦比亚血琼脂培养基选择性培养菌株,基质辅助激光解吸电离飞行时间质谱(MALDI⁃TOF⁃MS)系统进行菌种鉴定,微量肉汤稀释法检测菌株的最小抑菌浓度(MIC)。0.45μm滤膜过滤水样,富集样本,采用高通量宏基因组测序进行抗性基因多样性分析。【结果】从30件水样中分离到占比前5位的碳青霉烯多重耐药菌64株,分别为嗜麦芽窄食单胞菌(Stenotrophomonas maltophilia)、屎肠球菌(Enterococcus faecium)、铜绿假单胞菌(Pseudomonas aeruginosa)、鲍曼不动杆菌(Acinetobacter baumannii)和肺炎克雷伯菌(Klebsiella pneumoniae),这些菌株对多种临床常用的抗生素均耐药。通过宏基因组测序,检测到1244种抗性基因,前100个抗性基因的相对平均丰度占所有抗性基因96.1%,多重耐药类占63.41%,多重耐药基因主要为adeJ,mexT,adeC,oprM,mexF,mdfA,mexB,mdtK,adeK等。使用Spearman等级相关系数分析发现,水源地水中分离的5种多重耐药菌与抗性基因有明显正相关关系。【结论】该主要水源地原水中多重耐药基因相对丰度占比较高,5种分离到的碳青霉烯多重耐药细菌与抗性基因明显相关,涉及的抗生素种类与多重菌耐药表型相符。提示需持续加强抗生素的合理规范应用,保护水资源,保障饮水安全。[Objective]To determine the distribution of various antibiotic resistance genes(ARGs)in raw water from drinking water source,and to explore the correlation between the ARGs and common carbapenem-resistant and multidrug-resistant bacteria isolated from drinking water source,so as to provide scientific evidence for improving the safety of urban drinking water.[Methods]A total of 30 raw water samples were collected from a major drinking water source in Shanghai in 2020.Bacterial strains were selectively cultured on Columbia blood agar medium containing 1μg·μL^(−1)meropenem,and then identified by MALDI-TOF-MS mass spectrometry system.Minimum inhibitory concentration(MIC)of the strains was detected by broth microdilution method.The water samples were filtered through a 0.45μm filter membrane and diversity of ARGs was determined by using high-throughput metagenomic sequencing.[Results]A total of 64 strains of carbapenem-resistant bacteria were isolated from the water samples,including Stenotrophomonas maltophilia,Enterococcus faecium,Pseudomonas aeruginosa,Acinetobacter baumannii and Klebsiella pneumoniae,which were resistant to a variety of common antibiotics.Using metagenomic sequencing,1244 ARGs were identified.The relative average abundance of the top 100 ARGs accounted for 96.1%,and that of the multidrug-resistant ARGs accounted for 63.41%.Furthermore,the multidrug-resistant ARGs were mainly adeJ,mexT,adeC,oprM,mexF,mdfA,mexB,mdtK,adeK,etc.Using Spearman's correlation,five multidrug-resistant bacteria isolated from the drinking water source were significantly associated with the ARGs.[Conclusion]Relative abundance of multidrug-resistant ARGs is high in raw water from main drinking water source.The five isolated carbapenem-resistant and multidrug-resistant bacteria are significantly correlated with the ARGs.It warrants strengthening the rational and standardized application of antibiotics to protect water resources and ensure the safety of drinking water.

关 键 词：饮用水源地水 耐药细菌 抗性基因 多重耐药 高通量测序 

分 类 号：R123.7[医药卫生—环境卫生学]