利用交叉组装噬菌体示踪水环境中人源粪便来源的耐药细菌  

Tracing Antibiotic Resistant Bacteria from Human Feces in Water E nvironment Utilizing the Cross-assembly Phage

作　　者：段连瑞 段宇婧 李镇镇 苗泽丰 佘慧 杨涛 Duan Lianrui;Duan Yujing;Li Zhenzhen;Miao Zefeng;She Hui;Yang Tao(Department of Biochemistry and Molecular Biology,School of Basic Medicine,Shanxi Medical University,Taiyuan 030001,China;Higher Education Key Laboratory of Tumor Immunology&Targeted Drug Development in Shanxi Province,Shanxi Medical University,Taiyuan 030001,China;Key Laboratory of Coal Environmental Pathogenicity and Prevention,Ministry of Education,Shanxi Medical University,Taiyuan 030001,China;Department of Environmental and Safety Engineering,Taiyuan Institute of Technology,Taiyuan 030018,China)

机构地区：[1]山西医科大学基础医学院生物化学与分子生物学教研室,太原030001 [2]山西医科大学肿瘤免疫与靶向药物研发山西省高校重点实验室,太原030001 [3]山西医科大学煤炭环境致病与防治教育部重点实验室,太原030001 [4]太原工业学院环境与安全工程系,太原030018

出　　处：《生态毒理学报》2024年第4期100-111,共12页Asian Journal of Ecotoxicology

基　　金：国家自然科学基金青年项目(42107466);山西省基础研究计划(自由探索类)青年项目(20210302124498);山西省科技创新团队支持计划资助项目(202204051002030);山西医科大学校级博士启动基金项目(XD2110);山西省高等学校科技创新项目(2021L211)。

摘　　要：人类粪便中的耐药细菌(antibiotic resistant bacteria,ARB)及其携带的耐药基因(antibiotic resistance genes,ARGs)可以通过城市污水处理系统排放进入当地水环境,因此,快速准确地探明水环境中粪便污染及ARB情况对于保护生态系统和居民健康具有重要意义。本研究借助噬菌体crAssphage作为新型人类粪便污染特异性指示标记,采集太原市健康人群粪便、晋阳湖、汾河水库、自来水和污水处理厂的样品,首先采用实时荧光定量PCR技术检测样品中人类粪便污染指示标记crAssphage、细菌核糖体16S rRNA基因与耐药基因bla TEM-1的存在情况;其次针对各类样品进行细菌分离培养与鉴定,完成样品中bla TEM-1耐药菌株及多重耐药细菌的筛选;最后通过构建系统发育树分析菌株的进化关系,探究不同环境介质中细菌之间的相互影响。结果显示crAssphage在晋阳湖、汾河水库和污水处理厂样品中被检出,证明太原市区水环境存在人类粪便污染。本研究共分离鉴定出254株细菌,氨苄青霉素耐药细菌占比最高(74.41%),环丙沙星耐药细菌占比最低(1.97%);耐药基因bla TEM-1在各类样品的菌株中都有检出(22.83%),多重耐药细菌共有79株(31.10%)。进化关系分析表明人体肠道与水环境中的耐药细菌具有亲缘关系。上述结果表明太原市水环境受到人类粪便污染的影响,并且可能促进了ARGs和ARB的传播扩散,对人体健康产生威胁。该研究为评估太原市水环境粪便污染情况及ARB分布提供基础数据,为水环境监测保护给出科学依据。Antibiotic resistant bacteria(ARB)and the carrying antibiotic resistance genes(ARGs)in human feces can be discharged into the local water environment through the urban sewage treatment system.Therefore,it is of great significance to rapidly and accurately detect fecal pollution and ARB in water environment for safeguarding ecosystems and promoting public health.In this study,the bacteriophage crAssphage was utilized as a novel specific indicator for human fecal contamination.The samples were collected from feces of healthy individuals in Taiyuan,as well as water samples from Jinyang Lake,water reservoir,tap water and sewage treatment plant.Firstly,r eal-time fluorescent quantitative PCR(qPCR)was used to detect the indicator crAssphage,the presence of bacterial ribosome 16S rRNA gene and antibiotic resistance gene bla TEM-1 in all samples.Secondly,bacterial isolation,c ultivation and identification were carried out,followed by the screening of bla TEM-1 resistant strains and multidrug-resistant bacteria.Finally,the phylogenetic tree was constructed to analyze the evolutionary relationships among bacterial strains and explore the mutual interaction between bacteria in diverse environmental media.The results showed that crAssphage can be detected in samples collected from Jinyang Lake,water reservoir and sewage treatment plant,indicating the occurrence of human fecal pollution in water environment of Taiyuan.A total of 254 b acterial strains were isolated and identified in this study,with the highest proportion of ampicillin-resistant bacteria(74.41%)and the lowest proportion of ciprofloxacin-resistant bacteria(1.97%).The strains carrying antibiotic r esistance gene bla TEM-1 was also detected in all types of samples(22.83%),and 79 strains exhibited multidrug r esistance(31.10%).Evolutionary analysis revealed that there might exist a genetic relationship of ARB between the human gut and the water environment.The above results indicate that the water environment in Taiyuan is a ffected by human fecal pollution,potent

关 键 词：人类粪便污染 bla TEM-1 crAssphage 多重耐药菌 系统发育树 

分 类 号：X171.5[环境科学与工程—环境科学]