Mic r obial Sour c e T r ac king to battle E . c o li pollution in the Distr ic t of Columbia Amir S harifi, PhD Nov 21 2019 @ DOEE_DC
Pre se nta tio n o ve rvie w 1. Microbial Pollution in the District of Columbia 2. MS T basics 3. DOEE / EP A MS T Proj ect @DOE E _DC
F e c a l Po llutio n is a Na tio nwide Cha lle ng e Pathogens are the No.1 cause of impairments to the Rivers/ streams of the US . ht t ps:/ / ofmpub.epa.gov/ wat ers10/ at t ains_nat ion_cy.cont rol#causes @DOE E _DC
Mic ro b ia l Po llutio n in the Distric t o f Co lumb ia @DOE E _DC
Mic ro b ia l Po llutio n in the Distric t o f Co lumb ia • Blue Plains WTP and CS O (Combined ewer Overflow) ~95% bacterial load S • MS 4 (Municipal S eparate S torm S ewer S ystem) ~ 5% bacterial load @DOE E _DC
Co mmo n so urc e s o f E . c o li in MS4 • Illegal sanitary sewer connections to the storm drain • S anitary sewer exfiltration via groundwater seepage • Wildlife, such as birds and deer • Pets - especially dogs @DOE E _DC
DC Wa te r Qua lity Sta nda rds fo r E . c o li Water quality standards for E. coli in District specify that: • No single sample shall exceed 410 MPN/100 mL • The 30-day geometric mean should not exceed 126 MPN/100 mL ht t ps:/ / www.epa.gov/ wqs-t ech/ wat er-qualit y-st andards- regulat ions-washingt on-dc @DOE E _DC
So urc e o f F e c a l Po llutio n is I mpo rta nt • E. coli data alert to the total pollution level, however, does not provide information about the cause or source of pollutants • Public health risk can vary by source • Mitigation strategies can vary by source • S ource information improves water quality management and public safety @DOE E _DC
DNA-Ba se d Mic ro b ia l So urc e T ra c king • There are specialist microbes closely associated with a given pollution source o Host and gut microbes co-evolve Physiological differences of the gut Dietary differences between hosts • MS T provides a set of tools to characterize sources of contamination @DOE E _DC
Adva nta g e s o f q PCR fo r MST qPCR = quantitative Polymerase Chain Reaction Procedure for the measurement of host- associated gene sequences isolated from environmental water samples • “ Gold standard” for MS T • No cultivation requirement • Highly reproducible when standardized • Established quality control guidelines • S pecialized reagents for environmental testing @DOE E _DC
E PA Na tio na lly Va lida te d Me tho ds Dr. Orin Shanks U.S. EPA Office of Research & Development (ORD) @DOE E _DC
with Qua ntita tive re a l-time PCR (q PCR ) MST qPCR Data Sample DNA Sample Amplification Analysis Concentration Isolation Collection U.S. EPA- ORD Cincinnati, OH @DOE E _DC
DC MST Study Ob je c tive s 1. Characterize fecal source trends in select District outfalls to improve urban stormwater management 2. Develop procedure for future MS 4 outfall fecal pollution source characterization with MS T qPCR District EP A Region 3 National 13 @DOE E _DC
Se le c te d Site s East side of Anacostia River • • First order catchments • Size 13 to 123 ha • 32 MS4 outfalls across sites • Historic E. coli data (n = 202; since 2008) Chronic fecal pollution • (median E. coli geometric mean > 126 MPN/100ml) • No known CSO impact • Range of land use (100% parkland to 100% urban) • Dry weather MS4 outflows at most sites 14 @DOE E _DC
Sa mpling Pla n Dry weather • Precipitation 12-16 months • Flow Information 2x per month • Water Quality Metrics Receiving water MS 4 dry flow E. coli (IDEXX Colilert) outfalls Turbidity, Temperature, DO, pH • MS T qPCR Methods Wet weather Human-associated (HF183/ BacR287 and HumM2) 6-8 events Ruminant-associated (Rum2Bac) Receiving water MS Dog-associated (DG3) 4 dry flow outfalls only Avian-associated (GFD) @DOE E _DC
E vide nc e -Ba se d Hypo the sis T e sting H 1 : E. coli concentrations downstream of MS 4 outfalls will be higher during wet weather compared to dry weather conditions H 2 : Human sources will be more prevalent during dry weather outfalls compared to wet weather H 3 : Non-human sources will be more prevalent during wet weather outfalls compared to dry weather conditions H 4 : S patial and temporal trends will vary by pollution source and subwatershed land use practices @DOE E _DC
E . c o li mo nito ring , 7 e ve nts Low Density Parks Residential Fort Davis 54% 24% Texas Run 60% 18% https:/ / public.tableau.com/ profile/ amir.sharifi3095#!/ vizhome/ Graphs-mst/ Dashboard2 @DOE E _DC
E . c o li mo nito ring , 08-08-2019
E . c o li mo nito ring , 08-21-19 @DOE E _DC
E . c o li mo nito ring , 09-05-2019 @DOE E _DC
E . c o li mo nito ring , 09-18-19 @DOE E _DC
E . c o li mo nito ring , 10-02-19 @DOE E _DC
10-15-19 @DOE E _DC
E . c o li mo nito ring , 10-16-19 (we t we a the r e ve nt) 1.35” total rainfall @DOE E _DC
E . c o li mo nito ring , 10-31-19 @DOE E _DC
E . c o li mo nito ring , 11-14-19 @DOE E _DC
Ca se Study: T illa mook Ba sin, OR Applied human-associated qPCR methods (HF183/ BacR287 and HumM2), along with procedures for ruminant (Rum2Bac), cattle (CowM2 and CowM3), canine (DG3 and DG37), and avian (GFD) fecal pollution sources to characterize trends in fecal pollution sources in the research area. Li X, S ivaganesan M, KeltyCA, Zimmer-FaustA, ClintonP, ReichmanJR, et al. (2019)Large- @DOE E _DC scaleimplementationof standardizedquantitative real-timePCR fecal sourceidentificationprocedures in the TillamookBayWatershed.PLoS ONE 14(6):e0216827.https:/ / doi.org/ 10.1371/ j ournal.pone.0216827
@DOE E _DC
Avia n Pollution Spa tia l a nd T e mpora l T re nds Potential bird migration water quality impact Heat map: estimated log 10 copies per reaction color coding and frequency information.
0 Human Pollution Spatial and T e mpor al T r e nds River systems exhibit different temporal trends @DOE E _DC
Huma n F e c a l Co nta mina tio n Sc o re (HF S) Proj ect Level Probabilistic Modeling that can be used to prioritize sampling sites for remediation based on measured human waste levels. @DOE E _DC
Huma n F e c a l Co nta mina tio n sc o re (HF S) BMP performance can be assessed through HFS @DOE E _DC
Thank you for your attention! @DOE E _DC
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