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与水之间的一天或两天的延迟 样本收集和公共卫生决策。人们认识到，目前的这种做法并不能完全起到保护作用。 使用海水浴场的公众的健康。在这种情况下，可能需要几天时间才能 检测以前的粪便污染事件，重要的是了解粪便污染的类型 冲击着海滩。如果影响来自人类排泄物(如污水或化粪池)，情况尤其如此 因为这种排泄物污染源可能对人体健康构成重大风险。微生物科学 近年来，来源追踪技术的进步为检测粪便的性质提供了有效的新工具 海滩上的污染。我们将与我们的滑铁卢生物滤池系统(WBS)、多伦多公共卫生(TPH)和安大略省环境保护与园林部(MECP)合作伙伴一起，开发和测试新的监测和微生物来源跟踪工具，这些工具将加快加拿大重要分水岭多伦多的水质监测。我们将同时使用qPCR和高通量DNA测序进行DNA分析，以识别和量化多伦多市作为其定期海滩监测计划的一部分采集的污水样本中的特定细菌、病毒和原生动物。主要目标是量化肠球菌和大肠杆菌的水平，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)测试其他细菌(杆菌)和病毒(CrAssphage)粪便指标的存在；2)在DNA序列数据中对其他微生物进行生物信息学鉴定，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有DNA序列，我们将为WBS、MECP和TPH感兴趣的生物开发新的qPCR测试。这项研究的结果将提供全面的新信息和工具，可用于确保我们未来的供水质量。

Existing water quality monitoring protocols for E. coli are based upon a one or two day delay between water sample collection and public health decisions. It is recognized that this current approach is not fully protective of the health of the public using bathing beaches. Under these circumstances, where it may take several days to detect a previous fecal pollution event, it is important to have an understanding of the types of fecal pollution impacting a beach. This is particularly the case if the impacts are from human wastes (e.g. sewage or septic tanks), as this type of fecal pollution source can pose significant human health risks. The science of microbial source tracking has advanced in recent years to provide effective new tools for detecting the nature of fecal pollution 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.