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

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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.