Document QgOVV5GkZxNa55va5Q3D4O4d4

W i l i f c W I i l k1R I3 i l K l &2w m m 3 i 1 i I Sierra Club v. EPA 18cv3472 NDCA Tier 10 ED 002061 00176094-00001 m m Problem Swimming in recreational waters poses gastrointestinal illness risks to swimmers from fecal pollution -> need for improved monitoring tools in the toolbox (tens of millions of surface water associated swimming illnesses in the US a year resulting in billions of dollars in economic burden) - Direct detection of pathogens is a challenge - Pollution regulated with general fecal indicators Culture-based methods require overnight incubation Same day rapid methods improve public safety - Health risks originating from fecal sources are not equal Human and cattle fecal waste are of highest risk to humans - Bacterial indicators may not be suitable for viral pathogens which are believed to be the major cause of illness Sierra Club v. EPA 18cv3472 NDCA Tier 10 ED 002061 00176094-00002  w EFS^ Approach Recreational Water Quality Criteria (us era's office of water, 2012 update) ORD in Cincinnati focused on research in 3 areas: Rapid Quantitative Polymerase Chain Reaction (qPCR) methods for enterococci and E. coli fecal indicator bacteria qPCR microbial source tracking for human fecal contamination Coliphage (a type of bacteriophage, viruses that infect bacteria) as an alternative virus-based indicator of fecal contamination Sierra Club v. EPA 18cv3472 NDCA Tier 10 ED 002061 00176094-00003 oEPA Improve Methods for Detection of Bacterial General Fecal Indicators Development of rapid molecular methods for enterococci and E. coli Assess method robustness across the US Develop criteria for E. coli qPCR Provide implementation assistance for stakeholders Develop microbiological water quality forecasting tool (Virtual Beach) Virtual Beach is a software package designed by US EPA for developing site-specific statistical models for the prediction of pathogen indicator levels at recreational beaches. Available: 3 Sierra Club v. EPA 18cv3472 NDCA National Rivers and Streams Assessment Reprinted from US EPA Office of Water and ORD National Rivers and Streams Assessment 2008-2009: A Collaborative Survey (EPA/841/R-16/007). Washington, DC. March 2016. Tier 10 ED 002061 00176094-00004 v>EPA ' '' Microbial Source Tracking Innovative Methods Development of human-associated microbial source tracking qPCR methods Method selection by expert consensus Standardization of procedures Build laboratory capacity (training and standardized reagents) Science to support implementation Case Studies Great Lakes Beaches study Chicago Beaches study - Assessment at 9 beaches - Collaboration with University of Illinois at Chicago 4 Sierra Club v. EPA 18cv3472 NDCA Tier 10 ED 002061 00176094-00005 l^fl m m Coliphage Develop method to detect coliphage in ambient waters - Development/validation, performance across the US Determine occurrence in wastewater treatment and surface waters - Great Lakes Beaches study Fate and transport in wastewater treatment and environment - Comparisons to other indicators and viral pathogens S&j*#"IV - m&Mm Stem m i Sierra Club v. EPA 18cv3472 NDCA Tier 10 h* 'W& <*h\ gt? ED 002061 00176094-00006 Results: Improvements in enterococci and coli qPCR US EPA method 1609.1 (enterococci) and draft US EPA method C. (E. coli) - Validation studies for enterococci (completed) and E. coli (pending) Building stakeholder laboratory capacity - Training, collaboration with Michigan DEQ Development of DNA reference standards and standardization of methods Development of site specific E. coli qPCR criteria for beach notification - Collaboration with Michigan DEQ. Sierra Club v. EPA 18cv3472 NDCA m ': -' ' ' < ' \ . ' , ' . i m iw m xt k t!5,csm I inim m m . - y. y. . ____ ' .............................................................. v : . y . : v v y . 1 . ..........; . y . . . . - y : ..............................................................^ ........._ y ............... ittiis s : & y ; :::S>Sy: i- s ijw i f :# A K r f i i : .. y i 'y s . r f y * , Si:':!:;:*;. . !.i x .y v ? . V :< ? \ j; Tier 10 ED 002061 00176094-00007  EPA Results: Development and Validation of HumanAssociated Microbial Source Tracking Methods Human-associated qPCR methods for recreational water (for 2 types of bacteria, HF183 and HumM2) - Standardized procedure - Automated data analysis tool - Data acceptance metrics - Strong human-association - US EPA Region 5 lab training - Quantitative methods - Expert consensus - Field demonstrations - Implementation of tool by stakeholders - Michigan DEQ lab network training - Reference DNA material I ISCmahmparrpaolcevtseedrizIIaFti1o8n3oQfuHaunmtitaantivFeecRaelaPl-oTlliumtieonPCinRAAmssbaiyenfotrSurface Water Hvstt C.Green,* ktatd A. t uga >3,bMars11 Varrrs," Kars 7. Miller^ fvlark A. fcrcterdt^ Katherine G. Fiele!9V/illfernA. V isteti fi. Knight;9h Mano Sfgaresn*Catherine A Kfeltya OrinC. Shanks5 DSoautarcAe cIcdeepnttainficceatCiorniteQriuaanfotritaSttiavnedRaredailz-TedimHeuPmCaRn-MAsestohcoidasted Fecal OrinC. Shanks^a Catherine A. Kelty Robin OrhitO|b Sichard A. Hauglarr,5Tania ifedj5 Lauren Grooks,13Kat barine G. Ffeld,d Maro Svaganeana uo Sierra Club v. EPA 18cv3472 NDCA Tier 10 ED 002061 00176094-00008 wJTPHXWl^^'fWl mj^m Results: Coliphage Coliphage method development for fresh and marine surface waters and wastewater - Ultrafiltration to concentrate 1L samples followed by cultivation - Method validation (pending) - National assessment of method performance (pending) Coliphage fate and transport - Published results from OH and FL locations - Great Lakes Beaches study (pending) Sierra Club v. EPA 18cv3472 NDCA Tier 10 ED 002061 00176094-00009 ^EPA Building Stakeholder Laboratory Capacity US ERA Workshops arid Collaboration with Michigan PEQ coli and enterococci qPCR Training of labs on EPA Method 1609.1 for Enterococci Draft Method C for E. coil qPCR Multi-lab validation study on the E. coli method Study to assess the development of criteria for E. coli qPCR at Michigan Beaches and impact of applying enterococcus qPCR EPA Method 1609.1 Human associated source tracking markers Planning has been initiated for lab training on human-associated qPCR methods (HumM2 and HF183) Laboratory Training Course on EPA Metiiods 1609 and 1611 May 12 IS, 2014 Enterococci in Water fayTaqMan0 Quantitative Polymerase Chain Reaction (qPCR) - EPA An drew W. Breidenbaeh Envi ronrnental Researeh Center 9 26 W, Martin Luther King Drive Cincinnati,, OH 4S26S Sierra Club v. EPA 18cv3472 NDCA Tier 10 ED 002061 00176094-00010  w w^k. Indicator Occurrence: Great Lakes Field Studies (Ongoing) *Three Great Lakes beach areas *Water data collection: Enterococci and E. coli qPCR Enterococci and E. coli culture methods Microbial source tracking qPCR - Coliphage Edgewater Beach, Cleveland, OH * 16 water quality and recreational area parameters * Method comparisons Multiple publications FY 2018-19 10 Grant Park Beach, S. Milwaukee, Wl Washington Park Beach, Michigan City, IN Sierra Club v. EPA 18cv3472 NDCA Tier 10 ED 002061 00176094-00011 j l cIBhd'Mma m. Impact: Contributions to Recreational Water ' ' - - Criteria and Applications Toolbox Overall Impact * Improve microbiological water quality at beaches * Better inform when beaches should open or close qPCR for enterococci and E. coli * A tool that improves beach manager's ability to close and open beaches in a more timely manner. Enterococcus qPCR has a stronger relationship to Gl illness than overnight culture methods Microbial Source Tracking * Provides a tool for prioritization of beaches for remediation by identifying beaches that have the greatest human derived contamination Coliphage * A tool that targets viral indicators and has a potential to improve wastewater treatment and beach monitoring based on close relationship with viral pathogens 11 Sierra Club v. EPA 18cv3472 NDCA Tier 10 ED 002061 00176094-00012  w EFS^ Kevin Oshima US EPA Office of Research and Development National Exposure Research Laboratory Environmental Methods and Measurements Division Cincinnati, OH 513-569-7476 12 Sierra Club v. EPA 18cv3472 NDCA Tier 10 ED 002061 00176094-00013