Development and Evaluation of New Diagnostic Microbiological Tools for Monitoring Water Quality in Canadian Urban Environments

现有的大肠杆菌水质监测方案是基于水样收集和公共卫生决策之间的一到两天延迟。人们认识到，目前的做法并不能完全保护使用泳滩的公众的健康。在这种情况下，可能需要几天时间才能发现以前的粪便污染事件，因此了解影响海滩的粪便污染类型非常重要。如果影响来自人类废物（例如污水或化粪池），情况尤其如此，因为这类粪便污染源可对人类健康构成重大风险。近年来，微生物源追踪科学的发展为检测海滩粪便污染的性质提供了有效的新工具。我们将与我们的滑铁卢生物过滤系统（WBS）、多伦多公共卫生部（TPH）和安大略省环境、保护和公园部（MECP）合作伙伴一起，开发和测试新的监测和微生物源跟踪工具，这将加快多伦多这一加拿大重要流域的水质监测。我们将使用qPCR和高通量DNA测序进行DNA分析，以确定和量化多伦多市作为其定期海滩监测计划一部分的污水样本中的特定细菌，病毒和原生动物。主要目标将是量化肠球菌和大肠杆菌水平，以便迅速向公共卫生部门提供信息，以便于当天关闭海滩。其他研究目标将包括1)检测其他细菌（Bacteriodetes）和病毒（crASSphage）粪便指标的存在；2) DNA序列数据中其他微生物的生物信息学鉴定，包括细菌（弯曲杆菌、沙门氏菌、蓝藻）、病毒（冠状病毒- sars -2）和原生动物（隐孢子虫）。利用已鉴定的生物特异性DNA序列，我们将为WBS、MECP和TPH感兴趣的生物开发新的qPCR检测。这项研究的结果将提供全面的新资料和工具，以确保本港未来的供水质素。

Existing water quality monitoring protocols for E. coli are based upon a one or two day delay between watersample collection and public health decisions. It is recognized that this current approach is not fully protectiveof the health of the public using bathing beaches. Under these circumstances, where it may take several days todetect a previous fecal pollution event, it is important to have an understanding of the types of fecal pollutionimpacting a beach. This is particularly the case if the impacts are from human wastes (e.g. sewage or septictanks), as this type of fecal pollution source can pose significant human health risks. The science of microbialsource tracking has advanced in recent years to provide effective new tools for detecting the nature of fecalpollution at beaches. We will, in conjunction with our Waterloo Biofilter Systems (WBS), Toronto Public Health (TPH) and Ontario Ministry of the Environment, Conservation and Parks (MECP) partners, develop and test new surveillance and microbial source tracking tools that will expedite water quality monitoring in Toronto, an important Canadian watershed. We will conduct DNA analyses using both qPCR and high-throughput DNA sequencing to identify and quantify specific bacteria, viruses, and protozoa in water effluent samples taken by the City of Toronto as part of their regular, scheduled beach monitoring program. The primary goal will be to quantify Enterococcus and E. coli levels to quickly provide information to Public Heath to facilitate same-day beach closures. Additional research goals will include 1) testing for the presence of other bacterial (Bacteriodetes) and viral (crASSphage) fecal indicators; 2) bioinformatic identification of other microorganisms in DNA sequence data including bacteria (Campylobacter, Salmonella, Cyanobacteria), viruses (coronavirus-SARS-2) and protozoa (Cryptosporidium). Using identified organism-specific DNA sequences, we will develop novel qPCR tests for organisms of interest to WBS, MECP and TPH. The results of this study will provide comprehensive new information and tools that can be deployed to ensure the future quality of our water supply.