Mobile Water Kit, U of Alberta

Transcription

1 Pilot Testing of New Techniques for Illicit Discharge/Illicit Connection Mobile Water Kit Jeanne Dorn Canine Scent Tracking Debra Bouchard & Andrew Miller Human Bacteroides qpcr Debra Bouchard & Andrew Miller Mobile Water Kit, U of Alberta Developed for testing drinking water wells, rural India Mobile Water Kit King County Stormwater Services learned about MWK Pilot test set up between U of Alberta and King County Pilot test run June 2014; Capacity Grant funded Goal: assess usefulness as possible IC/ID detection tool Samples from four King County areas tested White Center Boise Creek North King County City of Kirkland Urban, piped system Ag-rural, ditches & stream Streams & lakes SW pipes, streams Boise1, June 2014 Sample location MWK filter 1

2 Mobile Water Kit ranges or bins Bin (& Bin number) E. coli, cfu/100 ml Very high (5) >10,000 High (4) 1000 to 10,000 Medium (3) 200 to 1000 Low (2) 50 to 200 Very low (1) 0 to 50 MWK & E. coli results, by bin number MWK & E. coli results (log scale) 2

3 MWK pilot test results General findings: Not there yet as a reliable tool for IC/ID in surface and stormwater Need to simplify kit for field use, and make it portable & durable Need to standardize & make easier the filter test color intensity interpretation Need to assure kit receptacle does not splash/cross-contaminate test filters White Center, June 2014 King County would be willing to conduct another pilot test on an improved version of the MWK These folk worked on MWK pilot test: King County: Sally Abella Bobbie Anderson Debra Bouchard Rebecka Carroll Cameron Chapman Maureen Dahlstrom Jeanne Dorn Joe Espinosa Bob Gilland Cynthia Hickey Jae Huh Rita Jordan Jennifer Keune Chris Knutson Terae Jones Tim O Leary Rebecca Love LeeAnn Merrill Doug Navetski Tim O Leary Susan Pratt Mark Preszler Jeanie Pride Dan Perzynski Mary Rabourn Joe Rochelle Dave Sizemore Ray Timm Debbie Turner City of Kirkland: Wes Ayers Jenny Gaus Ryean-Marie Tuomisto University of Alberta: Dr. Sushanta Mitra* Dr. Naga Siva Kumar Gunda* Hazel Robinson Ravi Shankar Chavali Venkata Subramanya * * Now at York University Canines Scent Tracking Human Bacteroides qpcr Microbial Source Tracking Tools Project Leads Debra Bouchard King County Science and Technical Support Katherine Bourbonais King County Environmental Lab Jonathan Frodge Seattle Public Utilities Ryean-Marie Tuomisto Kirkland Water Quality Karen Reynolds Environmental Canine Services 3

4 RUMINANT HUMAN CRITERIA Fecal coliform E. Coli Testing Microbial Source Tracking Tools Ruminant Bacteroidales qpcr (Rum-2-Bac) Human Bacteroides qpcr (Hu-2-Bac) Canine scent tracking Urban Watersheds Thornton Creek Juanita Creek Lake Washington Mathews Beach Juanita Beach Both Creeks Drain to swimming beaches with historically high fecal coliform Past intensive bacterial studies and partnerships No TMDL (yet) - Direct to Implementation Study Design Two samples collected at each site: Canine testing Lab testing E. coli / FC Hu-2-Bac Juanita Creek (73 samples) Thornton Creek (77 samples) Day 1 Day 2 Day 3 Day 4 Bucket test in scent neutral field site investigations site investigations Bucket test in scent neutral field site investigations site investigations 4

5 Controls in each basin (Day 1 & Day 3) 4 Positive Controls 4 Negative Controls 4 Alternate Waste Samples 2 Field Duplicates Influent diluted to 1%, 2%, 5%, and 10% Distilled H 2 0 1% to 2 % diluted feces Canine Source Tracking Human Source qpcr Hu2Bac qpcr = quantitative Polymerase Chain Reaction not a culturing process not relying on viable cells targets and quantifies the most common human gut Bacteroides spp top performing marker in 2013 study more specific than previous markers Sample membrane filtration -80 C aliquot processed reported in copies/ml 5

6 Canine and Lab Results Canines performed well on controls But did not agree with Hu-2-bac results Threshold of significance? Thornton Creek Results Thornton Creek Results 4,970, ,700 1,873, , ,100 24,900 9,996 31,000 Hu-2-bac (copies/100ml) 1,800 2,100 6

7 Thornton Creek Results 4,970, ,700 1,873, , ,100 24,900 9,996 31,000 1,800 2,100 Thornton Creek Results ,000 Pre-storm E. Coli (sampled 10/8/14) 10/13-14 Precip = 0.60 Post-storm E. Coli (sampled 10/14/14) 160 3, ,100 7, , , , ,400 4, , ,800 Juanita Creek Results 7

8 Juanita Creek Results 4,517 3,845 1, No sources found in follow-up investigations 1,192 2,851,404 Hu-2-bac (copies/100ml) Juanita Creek Results Juanita Creek Results , , , , ,100 12,000 8,000 Pre-storm E. Coli (sampled 10/8/14) 10/13-14 Precip = 0.60 Post-storm E. Coli (sampled 10/14/14) 8

9 Canine Scent Tracking Summary Pros Cons Rapid response for field work Control results were good one exception. May be useful on samples with FC levels 1% influent Variability between dogs Follow up dye and smoke testing showed nothing Uncertain what exactly in waste stream dogs key on (may not be fecal material) Did not correlate well with laboratory results Not local - cost $$$ for travel Human Bacteroides qpcr Summary Pros Control results were good Specific to human gut bacteria Quantitative results Can freeze samples for later analysis allowing for tiered processing Cons Interference can inhibit reading DNA can last in environment for up to a week depending on conditions Two week turnaround time Acknowledgements King County Andrew Miller Jeanne Dorn Katherine Bourbonais Colin Elliott Debbie Turner Stephanie Hess Eric Thompson Joe Clark Tami Alley Tim O Leary Doug Williams Logan Harris Seattle Public Utilities Jonathan Frodge Holli Brandt Angelique Hockett Nathan Hart Ellen Stewart Adam Bailey Environmental Canine Services: Karen Reynolds Gwen Molly s handler Aryn Crush s handler City of Kirkland: Ryean-Marie Tuomisto Wes Ayers Jenny Gaus Marie Jensen Most members of the Kirkland engineering and maintenance staff!! 9

10 Questions? King County Environmental Lab Services Human Organism or Gene Method and Result Units Type of fecal Marker Name Pollution Indicated Bacteroides spp. Membrane filtration & Real Indicates Possible Hu1Bac Time qpcr Human Source Cells/100 ml Bacteroides spp. Membrane filtration & Indicates Possible Hu2Bac qpcr Human Source Copies/ml Bifidobacteria spp. Culture Based Indicates Possible Human Source Cells/100 ml Performance Notes Cost for Cost for Prep Characteristics Complete and Hold Analysis Highly Sensitive 1 Presence $125 $50.00 (detects target at indicates low concentrations) recent human contaminatio n Top performing Presence $125 $50.00 human marker in indicates 2013 study 2 recent human contaminatio n High Confidence. Presence $175 N\A (Any level of indicates detection is recent significant) human contaminatio n Ruminant Rum2Bac Rhodococcus spp. Membrane filtration & Indicates Possible High Confidence. $125 $50.00 qpcr Ruminant Source Top performing ruminant marker in Copies/ml 2013 study 3 Culture Based Indicates Possible High Confidence $300 N\A Ruminant Source Cells/100 ml Costs here for single samples and may be discounted in larger batches. 10