Evaluation of Tracers of Fecal Pollution in Drinking Water Distribution Systems Walter Q. Betancourt and Minkyu Park Water and Energy Sustainable Technology Center. The University of Arizona RESEARCH PROGRESS REPORT UA-01-F2018 STATUS: Continuing, No Additional Funds Requested Relationship to Technology Roadmap: Municipal Water Use Practices and Wastewater Reuse Rationale: Approach: The Total Coliform Rule, a National Primary Drinking Water Regulation (NPDWR), uses Escherichia coli (E. coli) as a fecal indicator and total coliform (TC) to determine the adequacy of water treatment and the integrity of the drinking water distribution system (DWDS)   Host-specific bacterial gene sequences (genetic markers) and chemical compounds (fecal sterols) associated with humans and animals can be measured by highly sensitive analytical methods that provide more accurate and rapid (same-day) results than the culture-based method for E. coli and total coliforms Collect water samples from DWDS served by groundwater, 98% Central Arizona Project water, and water treated by advanced oxidation processes Apply Colilert, qPCR and GC-MS/MS for quantitative analysis of E. coli -TC, genetic markers and fecal sterols, respectively Human markers (qPCR): HF183, LACHNOS, C. coccoides, Blautia Ruminant fecal markers (CF193, Rum2Bac, CowM2 and CowM3) Avian fecal marker (GFD) Fecal sterols (GC-MS/MS): coprostanol, epicoprostanol, 24-ethylcoprostanol, and sitostanol Objectives: Key Deliverables: Compare analytical methods for microbial and chemical tracers of fecal pollution with the total coliform and E. coli assay Conduct quantitative analysis of genetic markers and fecal sterols in water distribution systems Develop a sensitive and reliable quantitative assay for rapid evaluation of drinking water safety in distribution systems Provide a sensitive and rapid (same-day results) approach to ascertain for fecal contamination in drinking water distribution systems based on the analysis of genetic markers and fecal sterols   Demonstrate the robustness of the microbial and chemical tracer analysis over the culture-based assay for total coliforms and E. coli to ascertain for drinking water safety in the distribution system Accomplishments: Development of all standard curves and methods for precise quantification of genetic markers by quantitative PCR assays (qPCR) and fecal sterols by gas chromatography-mass spectrometry (GC-MS/MS) Project Duration: July 1, 2018 – June 30, 2019 Water & Environmental Technology Center