Cyanobacteria, Toxins and Indicators. Field Monitoring Treatment Facility Monitoring Treatment Studies

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1 U.S. Environmental Projection Agency, Office of Research and Development Cyanobacteria, Toxins and Indicators Field Monitoring Treatment Facility Monitoring Treatment Studies Nicholas Dugan, Toby Sanan, Samantha Smith, Darren Lytle US Environmental Protection Agency Region 8 HABs Workshop Rapid City, SD September 30, 2015

Monitoring Treatment Studies Nicholas Dugan, Toby Sanan, Samantha Smith, Darren

2 Organization of Presentation Field monitoring Treatment plant influent monitoring Laboratory treatment studies

3 Office of Research and Development Field Monitoring

4 ELISA assay target Microcystin Molecule

5 Toxin Analysis Enzyme linked Immunosorbent assay (ELISA): Targets molecules of a specific shape in this case, the ADDA functional group Provides an approximation of total concentration of all microcystin congeners Liquid chromatography with tandem mass spectrometric detection (LC/MS/MS): Retention time Mass Fragmentation pattern Provides congener specific concentration data Only quantitates congeners for which standards are available

Liquid chromatography with tandem mass spectrometric detection (LC/MS/MS): Retention time Mass Fragmentation

6 Wyoming: Boysen Reservoir Swim beach Source: Wyoming Department of Environmental Quality

7 Boysen Boat Ramp

8 Region 8 Monitoring Results (State contingency samples) Site Extracellular ELISA (µg/l) Extra + Intracellular Extracellular LC/MS/MS (µg/l) Extra + Intracellular WY Boysen boat ramp 7/ WY Boysen swim beach 7/ < 0.05 WY Boysen treatment influent 7/29 < 0.25 < < 0.05 WY Boysen swim beach 9/2 < ND Homme dam 8/ ND Turtle Mountain site #1 9/1 31 > 50 > 5 > 400 ND Turtle Mountain site #2 9/1 30 > 120 > 6 > 600

1 230 WY Boysen swim beach 7/29 0.31 1.4 1.7 - < 0.05 WY Boysen treatment influent 7/29 < 0.25 < 0.25 - < 0.

9 Ohio River Bloom: September 2015 Source: Ohio River Sanitation Commission

10 Ohio River Monitoring Data (9/8/15 9/9/15) Microcystin concentration ( g/l) Medians: (1.5) (8.6) (2.0) Extracellular by ELISA Extra + intracellular by ELISA Extra + intracellular by LC/MS/MS

11 Lake Erie bloom: 6 August 2014 Sources: National Oceanic and Atmospheric Administration; National Aeronautics and Space Administration

12 Lake Erie Bloom: 10 August 2015 Sources: National Oceanic and Atmospheric Administration; National Aeronautics and Space Administration

13 Field Monitoring Takeaways Toxin concentrations are spatially heterogeneous across wide range of scales 1 ft to hundreds of miles Toxin concentrations are temporally heterogeneous Absolutely critical to monitor at the location of interest (beach, treatment facility intake, etc.) on a regular basis

concentrations are temporally heterogeneous Absolutely critical to monitor

14 Office of Research and Development Treatment Plant Intake Studies

15 Surface Water Treatment Process Coagulant Powdered activated carbon (PAC) Chlorine Permanganate Source Water Coagulation Sedimentation Flocculation Finished water Filtration Disinfection

16 Plant intake studies Two study areas: West four facilities in 2013, five facilities in 2014 East two facilities in 2013 & 2014 Toxin analyses: 2013 mostly by ELISA 2014 by ELISA and LC/MS/MS Chlorophyll a analyses: Proxy for suspended cyanobacterial biomass Other water quality analyses: Nitrate, nitrite, ammonia, phosphate

mostly by ELISA 2014 by ELISA and LC/MS/MS Chlorophyll a analyses: Proxy for

17 Lake Erie West study area East study area Source: National Oceanic and Atmospheric Administration

18 Chlorophyll a Summary (2013) Western study area Chlorophyll a Treatment plant influent (µg/l) Mean Minimum Maximum West West West West East East Eastern study area

20 4.2 66 West 2 31 0.89 102 West 3 11 0.17 40 West 4 17 1.

19 Inorganic Nitrogen at the Plant Intake (2013) (NH4+ + NO3- + NO2-) Nitrogen levels higher in western study area (red symbols) vs. eastern study area (green symbols) 4 4 Inorganic nitrogen (mg/l as N) West 1 West 2 West 3 West 4 East 1 East June July August September October November

eastern study area (green symbols) 4 4 Inorganic nitrogen (mg/l as N) West

20 Variable Bloom Dynamics The exact timing of the bloom peak can change from year to year 110 Chlorophyll ( g/l) West 2: 2013 bloom season West 2: 2014 bloom season Apr 0 May Jun Jul Aug Sep Oct Nov

2013 bloom season West 2: 2014 bloom season 100 90 90 80 80 70 70

21 Most of the cell removal work is accomplished prior to filtration May 19 July 1 July 21 Finished Filter eff pre Cl2 Clarifier eff Post PAC Post MnO Raw Chlorophyll a (ppb) Cell Removal at West 2 Treatment Plant (2014 Bloom Season)

22 Toxin Propagation Measured by ELISA (2014,West 2 Treatment Plant) Toxin Concentration by ELISA (µg/l) 120 Extracellular Toxin Total Toxin Intracellular toxin release following MnO4 addition Raw Post-MnO4 Post-PAC Filter Influent Filter Effluent Plant Tap Treatment Stage

23 Toxin Propagation Measured by LC/MS/MS (2014,West 2 Treatment Plant) Toxin Concentration by LC/MS/MS (µg/l) 10 Extracellular Toxin Total Toxin Intracellular toxin release following MnO4 addition 2 0 Raw Post-MnO4 Post-PAC Filter Influent Filter Effluent Plant Tap Treatment Stage

24 Toxin Speciation and Propagation Measured by LC/MS/MS Total Toxin Measurements Through Treatment by LC/MS/MS Extracellular Toxin Measurements Through Treatment by LC/MS/MS 3.5 MYC-LR MYC-RR MYC-YR MYC-LA MYC-WR MYC-LY Extracellular Toxin by LC/MS/MS (µg/l) Total Toxin by LC/MS/MS (µg/l) 10 MYC-LR MYC-RR MYC-YR MYC-LA MYC-WR MYC-LY Raw Post MnO4 Post PAC Filter Effluent Treatment Stage Finished Raw Post MnO4 Post PAC Filter Effluent Treatment Stage Significant diversity in toxin speciation More research needed to investigate effects of KMnO4 and PAC on toxin release/removal Finished

25 Office of Research and Development Laboratory Studies

26 Test Methodology Intact, toxin-producing cyanobacterial cells, grown in culture Cells suspended in de-chlorinated tap water Tests performed at ph 7 Potassium permanganate doses of 1, 2.5 and 5 mg/l Two trials at each permanganate dose ( A and B ) B trial with PAC addition at 30 minutes Tests performed in 2 L beakers on a standard jar test apparatus Samples collected prior to oxidant addition as well as at t = 15, 30 and 90 minutes Analyses conducted for total and extracellular toxins by ELISA

27 Toxin release: 1 mg/l KMnO4, ph 7 (ELISA) PAC addition correlates with an additional 2.3 µg/l removal of extracellular toxins

28 Toxin release: 2.5 mg/l KMnO4, ph 7 (ELISA) PAC addition correlates with an additional 5.9 µg/l removal of extracellular toxins

29 Toxin release: 5.0 mg/l KMnO4, ph 7 (ELISA) PAC addition correlates with an additional 0.1 µg/l removal of extracellular toxins

30 KMnO4 Concentrations (5 mg/l Dose) A fraction of the KMnO4 neutralized by PAC

31 Baseline to Peak Increase in Toxin Concentration Toxin release and degradation by KMnO4 happen concurrently KMnO4 Dose (mg/l) A Trial B Trial

32 Total Cell Counts Cyanobacterial cells maintain some degree of physical integrity during oxidation

33 Laboratory Test Highlights Increases in extracellular toxin concentrations observed at all doses Largest relative extracellular toxin increases (and lowest subsequent degradation) associated with lowest initial permanganate dose PAC addition correlates with decreases in extracellular toxin concentrations after 90 minutes No conclusive visual evidence of outright cell destruction

34 Disclaimer U.S. Environmental Protection Agency, through its Office of Research and Development, funded and managed, or partially funded and collaborated in, the research described herein. It has been subjected to the Agency s peer and administrative review and has been approved for external publication. Any opinions expressed in this paper are those of the author(s) and do not necessarily reflect the views of the Agency, therefore, no official endorsement should be inferred. Any mention of trade names or commercial products does not constitute endorsement or recommendation for use.

35 ThankYou! Nicholas Dugan

Cyanotoxin Removal in Drinking. Recreational Waters. Judy Westrick April 14, 2011 2011 Northeast Regional Cyanobacteria Workshop

Cyanotoxin Removal in Drinking Water Treatment Process and Recreational Waters Judy Westrick April 14, 2011 2011 Northeast Regional Cyanobacteria Workshop NEIWPCC Overview Water Treatment Source and Recreational