From the Headwaters To Boulder Creek Urban Corridor

Transcription

1 A Multifaceted Approach to Microbial Source Tracking Within Secondary Environments & Donna Scott Tetra Tech, Golden, CO City of Boulder Acknowledgments This project would not have been possible if not for the support of: Junko Munakata-Marr- CSM Larry Barber- USGS John Spear- CSM Donna Scott- City of Boulder Jill Tomaras - CSM Greg Brown- USGS Andrew Taylor - City of Boulder EPA GRANT #X General Outline Boulder Creek Watershed Background & Motivation Methods Alternative constituents Results - Comparison studies Recommendations From the Headwaters To Boulder Creek Urban Corridor Recreation Class 1A Primary contact Aquatic Life Class 1 1

2 QuickTime and a TIFF (Uncompressed) decompressor are needed to see this picture. Indicator Bacteria - Escherichia coli Presence indicates the water may cause illness. Five requirements of an indicator: Easy to detect. Generally not present in unpolluted waters. Concentrations can correlate to extent of contamination. Die-off rates are comparable to pathogens of concern. Generally not pathogenic Microbes in the Human Gut Fecal matter contains 1 12 bacteria cells per gram! That s approximately 9% of the feces weight! Of these bacteria, humans shed 2 x 1 9 coliforms/day/person (2,,,) Fecal coliforms: Only.1% of total microbes in feces! National Water Quality Inventory Stretch of Boulder Creek Impaired Water Quality Inventory: EPA. 2 33(d) listing Clean Water Act Impaired Waters Remediation Efforts - Best Management Practices established. Total Maximum Daily Load (TMDL) Program A tool for implementing state water quality standards. Based on the relationship between source of pollutants and in-stream water quality. Established allowable loading a water body can receive. TMDL: LC = WLA + LA + MOS 2

3 City of Boulder Outfalls Outfall Environments Dynamic & diverse Urban Runoff Motor oil Lawn care products Sediment Pet waste Plastics/garbage Toolbox Approach Utilize state of the art Multiple technologies Captures more complete view of system For wise men to go over, and fools to go under! Currently one method cannot answer all of the questions USEPA 25. The Truth About Poop By: Susan E. Goodman Wastewater Identification- Source Tracking Match microbe or chemical indicator from a polluted site to trace to the origin of pollution. Sources include: Treatment plants Septic systems Agricultural runoff Wildlife Boulder Creek Methods Bacterial IDEXX: E. coli Real-time, quantitative Polymerase Chain Reaction (PCR): Bacteroides (HF-183) Chemical Fluorescence: Optical brighteners ELISA: Enzyme-Linked Immunosorbant Assays GC-MS: Gas Chromatography-Mass Spectrometry 3

4 Boulder Creek E. coli Daily Geometric Mean (log CFU/1mL) E. coli Concentrations (Geometric Mean CFU/1mL) E. coli Geometric Mean (cfu/1ml) Orodell Gauge Daily Mean Discharge cfs 3/1/27 4/1/27 5/1/27 6/1/27 7/1/27 8/1/27 9/1/27 1/1/27 11/1/27 12/1/27 1/1/28 2/1/28 3/1/28 4/1/28 5/1/28 6/1/28 7/1/28 8/1/28 9/1/28 1/1/28 11/1/28 12/1/28 1/1/29 2/1/29 3/1/29 4/1/29 5/1/29 6/1/29 7/1/29 Average daily water temperature (C) Boulder Creek -- Monitoring Goals 7 Boulder Creek Daily E.coli, flow & water temperature 3 1. Identify concern temporal & spatial. Target Sampling Boulder Creek daily geomean 2. Examine persistence & background levels of E. coli. 3. Implement multiple analyses to accurately identify wastewater contamination. Toolbox Approach Orodell gauge daily mean discharge (cfs) Average water temps (C) 3 Boulder Creek 27 Targeted Sampling Temporal Distribution Impairment specific to lower portion January February March April May June July August Septmeber October November December Boulder Creek Environmental Trends E. coli vs Temperature Recent E. coli Research Persist within the aquatic environment, soils and substrate (Byappanahalli 23) y =.37x R² =.42 Soil exhibited cohesive distinct phylogenic groups (Byappanahalli 26) Water Temperatures (C) Controlled by predation direct transfer from bacterial mass to predation growth. (Jurgens et al. 1997). 4

5 Water column E. coli concentrations (log CFU/1mL) Persistence Research- Environmental Contribution? Boulder Creek E. coli Concentrations Sediment vs Water Column y =.8547x R² = Sediment E. coli concentrations (log CFU/1mL) Boulder Creek Benchtop Sediment Microcosms E. coli Genetics E.coli Concentrations (CFU/1mL) /14 1/3 11/7 11/24 12/1 Date 12/18 1/5 1/21 Variable BC CAN BC CU OUT POM OUT SKI Whittam addressed impacts of natural selection on environmental strains (1989). Primary vs Secondary environments Low number of virulent and antibiotic resistance strains, suggesting characteristics could be lost in the transition from primary to secondary environment. (Kon et al. 27) Source Tracking Guide Source Tracking of Wastewater Microbial Source Tracking Overview Bacterial Chemical E. coli Bacteriodes Optical Brighteners (OB) ELISA Triclosan & 17B GC-MS Trace Organics Graphics by: Jill Tomaras 5

6 Bacteroides (log #/ml) E. coli stakeholder meeting Alternative Indicators - Library Dependent? Microbial: The primary detection method for microorganisms is via culturing: using growth medium in a test tube or dish However, 99.9% of microbes are unculturable! Why Bacteroides? Only found in feces, rumen, and body cavities 1/3 of fecal flora Anaerobe - Limited survival in environment Host-specific variation in animal host Found to correlate more often than E. coli to pathogens (Savichtcheva 27). Bacteroides - Research & Method Development Boulder Creek Bacteroides qpcr Don Stoeckel - USGS Bernhard & Dick Field Seurinck Ahmed McQuaig Boulder Creek E. coli vs Bacteroides y =.1551x R² =.31-4 Concentration of E. coli (log CFU/1mL) 15 significant detects. 1 positive E. coli correlated with 1 positive Bacteroides. 7 positive E. coli NOT associated with Bacteroides. Only positive Bacteroides w/o E. coli at WWTP. Source Tracking Guide Source Tracking of Wastewater Bacterial Chemical Alternative chemical indicators Alternative chemical indicators of wastewater: E. coli Bacteriodes Optical Brighteners (OB) ELISA Triclosan & 17B GC-MS Trace Organics 1. Personal Care Products 2. Pharmaceuticals 3. Hormones 4. Metabolites 6

7 QuickTime and a decompressor are needed to see this picture. QuickTime and a decompressor are needed to see this picture. Optical Brighteners What is an FWA/OB? Relative Fluorescence Need to quantify levels. Standards most common FWA (Tenopal CBS-X) USGS, Boulder, CO Larry Barber Photo: Hartel Photo: Hartel Relative Fluorescence ELISA (Enzyme-Linked Immunosorbant) Competitive reaction Fluorescence Triclosan 17 B Estradiol ELISA Results Triclosan Antimicrobial Resistant strains Levels found within: Outfalls: -343ng/L In-stream: 34.46ng/L 17 -Estradiol Naturally occurring hormone. Estrogenic in nature Levels found within: Outfalls: -38.8ng/L In-stream: ave 3.3ng/L Solid Phase Extraction Surrogate Standards Full scan for standards SIM scan for samples Peer reviewed Gas Chromatography - Mass Spectrometry 7

8 GC-MS Detects Bisphenol-A 4-Methylphenol 4-tert-butylphenol N,N,diethyl-m-toluamide (DEET) 4-n-octylphenol Caffeine Triclosan GC-MS - Caffeine Levels found within: Outfalls: ,ng/L In-stream: 42ng/L 4-Methylphenol Disinfectant Solvent Levels found within: Outfalls: -256ng/L In-stream: 19ng/L GC-MS Detects Bisphenol-A Plasticizer PVC piping Toilet paper Endocrine disruptor Levels found within: Outfalls: 9-24ng/L In-stream: 38ng/L Bacterial E. coli Bacteriodes Source Tracking of Wastewater Chemical Optical Brighteners (OB) ELISA GC-MS Environmental Trends? Comparison #1: Out-Pom E. coli 26.2 CFU/1mL TOC 89mg/L OB 98ug/L Caffeine 12,275ng/L Triclosan 95ng/L ng/L Comparison #2: Out-Ski E. coli 7,71 CFU/1mL Bacteroides.1copies/mL TOC 1.8mg/L OB 45ug/L Caffeine ng/l Triclosan 1ng/L 17-.9ng/L 8

9 Comparison #3: Out-Fol E. coli 9,84 CFU/1mL Bacteroides 34 copies/ml OB 45ug/L Caffeine 297ng/L Triclosan 31ng/L ng/L What it all means. Relationships between indicators could not be established. Does not weaken alternative indicators Raises further doubt in the utility of E. coli as an indication of wastewater contamination. Due to environmental persistence, E. coli is not completely accurate in identifying recent contamination. Multiple constituents must be used in order to accurately detect a broad range of humanderived contamination. WERF 29 What s next TMDL development The knowledge gaps are so large and the inherent variability in NPS-impacted streams is so great that we do not know if the water quality criteria based on Great Lakes/coastal studies can be extrapolated to inland waters. These criteria could either be over- or underprotective of human health WERF Inland Waters Workshop Report, 29, pg The E. Coli TMDL Quandary E. coli poses a number of challenges for stormwater managers Questionable indicator of pathogens Unknown public health risk Sources may be diffuse and difficult to identify and control Difficult to quantify TMDL loads from nonpoint sources Hard to develop/implement BMPs Identifying Sources of E. coli Storm drains (WLA): Inconsistent flow Sporadic events Variable catchments 9

10 Sources of E. coli Sanitary Sources Sources of E. coli Diffuse and Difficult to Identify and Control The Wildlife Exclusion? Sewer overflows, leaking pipes, infiltration, etc First Priority: Identify and eliminate sanitary sources Leave No Trace 1999 study: Dog Excrement 68 pooping events were observed. Excrement picked up 59% Excrement left 41% Diffuse and Difficult to Identify and Control What can we do? Stormwater BMPs - Sediment Boulder Shelter for the Homeless assisting over 1 people per year. Recreational Sources -kayaking, tubing, tributary path system users. BMPs - End of Pipe Treatment? Non-structural BMPs Public Education Costly Questionable effectiveness Worst case treat specific problem outfalls, capture first flush Hard to quantify effectiveness 1

11 Is TMDL the best regulatory option? E. coli is not a good indicator of pathogens and their health risk difficult to quantify, model and to calculate loading rates numeric standard may not be appropriate for this constituent If not TMDL BMP requirements added to MS4 permits Identify and Remove Sanitary Sources outfall inventory in-stream and outfall pathogen and wastewater constituent screening and monitoring Additional BMPs to target constituents and reduce public health risk Future Recommendations: Need for source tracking All Bac (Total Bacteroides) Annual sampling Persistence - level of constituents Human metabolites - additional constituents Questions? 11