庄芳芳;苏建强;陈辉煌;朱永官;

• 1:中国科学院城市环境研究所城市环境与健康重点实验室
• 2:中国科学院大学

摘要(Abstract)：

水体微生物污染(包括致病菌、病毒、寄生虫)会引起多种传染病和寄生虫病,对生产生活用水安全和人体健康造成重要威胁。本研究应用基于Taq Man探针的高通量荧光定量PCR技术对厦门市后溪流域冬季微生物污染进行检测,包含了5种粪便污染源(人源、反刍动物源、猪源、家禽源、狗源)微生物源示踪分子标记物与12种病原微生物。结果表明,该流域在上游及水库5个位点没有粪便污染,仅在其中一个水库位点检测出棘阿米巴,微生物污染极小;中下游检测出人类、反刍动物、猪、家禽、狗粪便污染,并且检测出产气荚膜梭菌、肠聚集性大肠杆菌、肠毒素型大肠杆菌、幽门螺杆菌、霍乱弧菌、副溶血弧菌、棘阿米巴、克雷伯氏肺炎杆菌等病原菌,其中流经旧城区居民生活生产区水样微生物污染严重,下游新城区微生物污染较小。这些结果暗示着城市人类活动是流域微生物污染主要来源,应从污染源头加强微生物污染控制。

关键词(KeyWords)： 流域;微生物污染;微生物源示踪;病原微生物;高通量荧光定量PCR

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(31722004);; 国家重点研发计划重点专项(2016YFD0800205)

作者(Author): 庄芳芳;苏建强;陈辉煌;朱永官;

Email:

参考文献(References)：

• [1]杨勇,魏源送,郑祥,等.北京温榆河流域微生物污染调查研究[J].环境科学学报,2012,32(1):9-18Yang Y,Wei Y S,Zheng X,et al.Investigation of microbial contamination in Wenyu River of Beijing[J].Acta Scientiae Circumstantiae,2012,32(1):9-18(in Chinese)
• [2]国家环境保护总局.GB3838—2002地表水环境质量标准[S].北京:中国标准出版社,2002Ministry of Environmental Protection of the People's Republic of China.GB3838—2002 Environmental quality standards for surface water[S].Beijing:China Standard Press,2002(in Chinese)
• [3]世界卫生组织.饮用水水质准则(第四版)[M].上海:上海交通大学出版社,2014:87-232World Health Organization.Guidelines for Drinking-water Quality(Fourth Edition)[M].Shanghai:Shanghai Jiaotong University Press,2014:87-232(in Chinese)
• [4]Simpson J M,Santo D J,Reasoner D J.Microbial source tracking:State of the science[J].Environmental Science and Technology,2002,36(24):5279-5288
• [5]王耀兵,苏洁,杨玉敏,等.一种粪便污染源识别新技术——微生物源示踪[J].海洋环境科学,2008,27(S2):122-128Wang Y B,Su J,Yang Y M,et al.New technology of identification of fecal pollution——Microbial source tracking[J].Marine Environmental Science,2008,27(S2):122-128(in Chinese)
• [6]段传人,刘爱喜,王贵学,等.应用肠道微生物示踪地表水粪便污染的研究进展[J].微生物学通报,2013,40(12):2319-2329Duan C R,Liu A X,Wang G X,et al.Research progress of using intestinal microbes to track the sources of fecal pollution in surface water[J].Microbiology China,2013,40(12):2319-2329(in Chinese)
• [7]Wiggins B A.Discriminant analysis of antibiotic resistance patterns in fecal streptococci,a method to differentiate human and animal sources of fecal pollution in natural waters[J].Applied and Environmental Microbiology,1996,62(11):3997-4002
• [8]Parveen S,Murphree R L,Edmiston L,et al.Association of multiple-antibiotic-resistance profiles with point and nonpoint sources of Escherichia coli in Apalachicola Bay[J].Applied and Environmental Microbiology,1997,63(7):2607-2612
• [9]Scott T M,Rose J B,Jenkins T M,et al.Microbial source tracking:Current methodology and future directions[J].Applied and Environmental Microbiology,2002,68(12):5796-5803
• [10]Harwood V J,Staley C,Badgley B D,et al.Microbial source tracking markers for detection of fecal contamination in environmental waters:Relationships between pathogens and human health outcomes[J].FEMS Microbiology Reviews,2014,38(1):1-40
• [11]魏源送,郑嘉熹,王光宇,等.地表水微生物溯源技术的开发和应用进展[J].水资源保护,2016,32(1):1-11Wei Y S,Zheng J X,Wang G Y,et al.Development and application progress of microbial source tracking in surface water[J].Water Resources Protection,2016,32(1):1-11(in Chinese)
• [12]陈亚楠,王亚炜,魏源送,等.不同功能地表水体中病原微生物指示物的标准比较[J].环境科学学报,2015,35(2):337-351Chen Y N,Wang Y W,Wei Y S,et al.Evolution and standard comparison of indicator microorganisms for different surface waters[J].Acta Scientiae Circumstantiae,2015,35(2):337-351(in Chinese)
• [13]Field K G,Samadpour M.Fecal source tracking,the indicator paradigm,and managing water quality[J].Water Research,2007,41(16):3517-3538
• [14]Ferguson A S,Layton A C,Mailloux B J,et al.Comparison of fecal indicators with pathogenic bacteria and rotavirus in groundwater[J].Science of the Total Environment,2012,431:314-322
• [15]张崇淼,王晓昌,周进宏,等.城市地表水中肠道病原微生物与粪便污染指示菌的关系研究[J].环境科学学报,2012,32(11):2789-2794Zhang C M,Wang X C,Zhou J H,et al.Comparative study on enteric pathogens and their relations with fecal indicator bacteria in urban surface waters[J].Acta Scientiae Circumstantiae,2012,32(11):2789-2794(in Chinese)
• [16]王宁,黄友谊,陈伟伟.构建城市水系生态安全格局初探——以厦门市后溪为例[C].青岛:城市时代,协同规划——2013中国城市规划年会,2013:1-12Wang N,Huang Y Y,Chen W W.A Preliminary study on establishing an ecological security pattern of urban water system—A case study of Houxi River,Xiamen[C].Qingdao:City Times,Collaborative Planning-China Urban Planning Annual Meeting 2013,2013:1-12(in Chinese)
• [17]Liu L,Yang J,Yu X,et al.Patterns in the composition of microbial communities from a subtropical river:Effects of environmental,spatial and temporal factors[J].PLOS One,2013,8(11):e81232
• [18]Mieszkin S,Furet J P,Corthier G,et al.Estimation of pig fecal contamination in a river catchment by real-time PCR using two pig-specific Bacteroidales 16S r RNA genetic markers[J].Applied and Environmental Microbiology,2009,75(10):3045-3054
• [19]Shanks O C,Kelty C A,Sivaganesan M,et al.Quantitative PCR for genetic markers of human fecal pollution[J].Applied and Environmental Microbiology,2009,75(17):5507-5513
• [20]Lee C S,Lee J.Evaluation of new gyr B-based real-time PCR system for the detection of B.fragilis as an indicator of human-specific fecal contamination[J].Journal of Microbiological Methods,2010,82(3):311-318
• [21]Green H C,Haugland R A,Varma M,et al.Improved HF183 quantitative real-time PCR assay for characterization of human fecal pollution in ambient surface water samples[J].Applied and Environmental Microbiology,2014,80(10):3086-3094
• [22]Schill W B,Mathes M V.Real-time PCR detection and quantification of nine potential sources of fecal contamination by analysis of mitochondrial cytochrome b targets[J].Environmental Science&Technology,2008,42(14):5229-5234
• [23]Layton A,Mckay L,Williams D,et al.Development of Bacteroides 16S r RNA gene Taq Man-based real-time PCR assays for estimation of total,human,and bovine fecal pollution in water[J].Applied and Environmental Microbiology,2006,72(6):4214-4224
• [24]Kobayashi A,Sano D,Hatori J,et al.Chicken-and duckassociated Bacteroides-Prevotella genetic markers for detecting fecal contamination in environmental water[J].Applied Microbiology and Biotechnology,2013,97(16):7427-7437
• [25]Kildare B J,Leutenegger C M,Mcswain B S,et al.16S r RNA-based assays for quantitative detection of universal,human-,cow-,and dog-specific fecal Bacteroidales:A Bayesian approach[J].Water Research,2007,41(16):3701-3715
• [26]Green H C,White K M,Kelty C A,et al.Development of rapid canine fecal source identification PCR-based assays[J].Environmental Science and Technology,2014,48(19):11453-11461
• [27]Wroblewski D,Hannett G E,Bopp D J,et al.Rapid molecular characterization of Clostridium difficile and assessment of populations of C.difficile in stool specimens[J].Journal of Clinical Microbiology,2009,47(7):2142-2148
• [28]Lee D Y,Shannon K,Beaudette L A.Detection of bacterial pathogens in municipal wastewater using an oligonucleotide microarray and real-time quantitative PCR[J].Journal of Microbiological Methods,2006,65(3):453-467
• [29]Liu J,Gratz J,Amour C,et al.A laboratory-developed Taq Man Array Card for simultaneous detection of 19 enteropathogens[J].Journal of Clinical Microbiology,2013,51(2):472-480
• [30]Kobayashi D,Eishi Y,Ohkusa T,et al.Gastric mucosal density of Helicobacter pylori estimated by real-time PCR compared with results of urea breath test and histological grading[J].Journal of Medical Microbiology,2002,51(4):305-311
• [31]Bliem R,Schauer S,Plicka H,et al.A novel triplex quantitative PCR strategy for quantification of toxigenic and nontoxigenic Vibrio cholerae in aquatic environments[J].Applied and Environmental Microbiology,2015,81(9):3077-3085
• [32]Wang H,Edwards M,Falkinham J R,et al.Molecular survey of the occurrence of Legionella spp.,Mycobacterium spp.,Pseudomonas aeruginosa,and amoeba hosts in two chloraminated drinking water distribution systems[J].Applied and Environmental Microbiology,2012,78(17):6285-6294
• [33]Su J Q,Wei B,Ou-Yang W Y,et al.Antibiotic resistome and its association with bacterial communities during sewage sludge composting[J].Environmental Science&Technology,2015,49(12):7356-7363
• [34]Zhu Y G,Zhao Y,Li B,et al.Continental-scale pollution of estuaries with antibiotic resistance genes[J].Nature Microbiology,2017,2:16270
• [35]路凤,金银龙,程义斌.军团菌病的流行概况[J].国外医学(卫生学分册),2008(2):78-83Lu F,Jin Y L,Cheng Y B.Epidemiology of Legionella disease[J].Foreign Medicine(Hygiene),2008(2):78-83(in Chinese)
• [36]James E S,Joyce M P.Assessment and management of watershed microbial contaminants[J].Critical Reviews in Environmental Science and Technology,2004,34(2):109-139
• [37]Blackstone G M,Nordstrom J L,Bowen M D,et al.Use of a real time PCR assay for detection of thectx A gene of Vibrio cholerae in an environmental survey of Mobile Bay[J].Journal of Microbiological Methods,2007,68(2):254-259