Preliminary Assessment of Cyanotoxin Occurrence in Lakes and Reservoirs in the

United StatesKeith A. Loftin, Jennifer L. Graham, Michael T. Keith A. Loftin, Jennifer L. Graham, Michael T.

Meyer, Andrew C. Ziegler, Julie E. DietzeMeyer, Andrew C. Ziegler, Julie E. DietzeU.S. Geological SurveyU.S. Geological Survey

SSusan Holdsworth, Ellen Tarquiniousan Holdsworth, Ellen TarquinioU.S. Environmental Protection AgencyU.S. Environmental Protection Agency

Acknowledgements

USEPA Office of Wetlands, Oceans and Watersheds (National Lake Assessment Website: http://www.epa.gov/owow/lakes/lakessurvey/)

USGS Toxic Substance Hydrology Program (http://toxics.usgs.gov/regional/emc/)

Disclaimer:

Any use of trade, product, or firm names in this presentation is for descriptivepurposes only and does not imply endorsement by the U.S. Government.

A. Concerns and Guidelines

B. Introduction to Cyanotoxins and Cyanobacteria

USGS cyanotoxin sampling protocol:

http://pubs.usgs.gov/sir/2008/5038

C. Previous Occurrence in Midwest 1999-2006

D. Toxin Studies

1. 2007 EPA National Lake Assessment (2007 US EPA NLA)

2. 2006 USGS Midwestern US Cyanotoxin Reconnaissance of

Cyanobacterial Blooms (2006 USGS MCR)

E. Take Home Message

Outline

Lakeline, 26(2), Summer 2006.

Concerns and Guidelines

• Known Acute effects and chronic effects- hepatotoxins- neurotoxins- dermatotoxins- carcinogens

• EPA Contaminant Candidate List 3• Anatoxin-a• Cylindrospermopsin• Microcystin-LR

• WHO guidelines – Microcystin-LR• Drinking water – 1.0 ppb• Recreational water

– Low Risk - < 10 ppb– Moderate Risk - 10-20 ppb– High Risk - > 20 ppb

Binder Lake, IA August 2006

Newsworthy Cyanotoxin Events in the U.S.

At Least 35 U.S. States With At Least 35 U.S. States With ReportsReports

of Cyanotoxin Associated of Cyanotoxin Associated PoisoningsPoisonings

= reported incident

Cyanobacteria Made the News in at Cyanobacteria Made the News in at Least 21 U.S. States During 2006Least 21 U.S. States During 2006

= news report

East Okoboji Lake, IA June 2000

After Graham, 2006

Photic Zone Sampling vs. Bloom SamplingPhotic Zone Sampling vs. Bloom Sampling

Upper Gar, IA August 2006Upper Gar, IA August 2006

Cyanotoxin transport:Intracellular:

WindHydraulic Bouyancy Regulation

Extracellular:Hydraulic (Dissolved-Phase)Particulate-Bound

Sampling protocol critical to ability to interpret data!

Higher concentrations of cyanobacteria and toxins more likely in wind blown accumulations near shore.

Cheney Reservoir, KS, June Cheney Reservoir, KS, June 20032003

Environmental Distribution of CyanotoxinsEnvironmental Distribution of Cyanotoxins“The Quick Version”“The Quick Version”

ToxinToxin

IntracellularToxin

DissolvedToxin

SorbedToxin

IntracellularToxin

DissolvedToxin

SorbedToxin

Dissolved-Phase Toxin (Extracellular)

= +

Particulate Toxin – meaningdepends on whether cells are lysed.

Total Toxin

Total Toxin = Extracellular Toxin + Intracellular Toxin + Particulate Toxin

Microcystins were Commonly Found in the Midwest by ELISA (1999 – 2006)

After Graham and others 2004 and 2006

1. Integrated photic zonesamples.

2. 78% (n = 359) of lakes had at least one detectable occurrence of toxins.

3. Total concentrations ranged from:

< 0.10 to 52 ppb.

Mozingo Lake, MO October 2001

Randomly selected lakes for 2007 EPA Randomly selected lakes for 2007 EPA National Lake Assessment - National Lake Assessment - Photic Zone SamplingPhotic Zone Sampling

Image from EPA 841-F-06-002

(http://www.epa.gov/owow/lakes/lakessurvey/)Total lakes: n=1150 for Microcystins

1. Total microcystin samples collected in photic zone in deepest part of lake.

2. All samples were analyzed by ELISA.

3. 2 % of samples willbe analyzed by LC/MS/MS forconfirmation.

2007 EPA NLA – Photic Zone Sampling 2007 EPA NLA – Photic Zone Sampling Microcystins Were Found Throughout the United States Microcystins Were Found Throughout the United States

Especially in the Upper Midwest.Especially in the Upper Midwest.

% Overall Detections (with Reference Lakes andResampled Lakes): 32 % (401/1238)

Total Microcystin Concentration (ppb MCLR equivalents) Study Lakes

Mean 3.0 (~ 1.0)Median 0.52 (< 0.10)Minimum 0.10 (< 0.10)Maximum 230

1 Values outside parenthesis are summary statistics for detections only. Values inside parenthesis include non-detections in summary statistics.

2006 USGS Midwestern US Cyanotoxin 2006 USGS Midwestern US Cyanotoxin Reconnaissance – Targeted Cyanobacterial Blooms Reconnaissance – Targeted Cyanobacterial Blooms

and Accumulationsand Accumulations

1. Cyanobacterial blooms and accumulations in 23 midwestern lakes (KS, MO, IA, MN) were targeted over a 1 week period inAugust 2006.

2. Measurements included: water chemistry, cyanotoxin analysis by ELISAs and LC/MS/MS (dissolved and total), taste-and-odor compounds (SPME-GC/MS), chlorophyll a (fluorescence), and phytoplankton ID.

2006 USGS Midwest US Cyanotoxin Reconnaissance During August 2006 all lakes (n=23) had detectable microcystins when ELISA and LC/MS/MS

are combined, 30% had detectable Anatoxin-a, and 9% had detectable Nodularin-R.

ELISA (MRL = 0.10 ppb, 0.02 ppb, 0.04 ppb)

1. ELISA detections of Cylindrospermopsin were not reproduced by LC/MS/MS2. ELISA detections for Saxitoxins could not be confirmed at this time. Still

working to include these compounds in our LC/MS/MS method.

LC/MS/MS (MRL = 0.010 ppb)

2006 USGS Midwest Recon 2006 USGS Midwest Recon Microcystins Were the Dominant Class of CyanotoxinsMicrocystins Were the Dominant Class of Cyanotoxins

(ELISA - 23 lakes) (ELISA - 23 lakes)

Upper Pine Lake, IA August 2006Upper Pine Lake, IA August 2006 Rock Creek Lake, IA August 2006Rock Creek Lake, IA August 2006

Total Concentration (ppb) Cylindrospermopsins

Microcystins and Nodularins Saxitoxins

Mean 0.13 (0.011) 1 600 (570) 0.07 (< 0.02)Median 0.13 (< 0.04) 3.0 (2.8) 0.03 (< 0.02)Minimum < 0.04 < 0.10 < 0.02Maximum 0.14 13000 0.20

1 Values outside parenthesis are summary statistics for detections only. Values inside parenthesis include non-detections in summary statistics.

2006 USGS MCR – Blooms and Accumulations2006 USGS MCR – Blooms and AccumulationsMicrocystins –LR and –RR were the Most Abundant Congeners and Microcystin–RR had Microcystins –LR and –RR were the Most Abundant Congeners and Microcystin–RR had

the Highest Concentrations.the Highest Concentrations.

No detections above 0.010 ppb for Cylindrospermopsin, Deoxycylindrospermopsin, and Lyngbyatoxin-A.

30%% Detects 52% 39% 91% 39% 61% 78% 65% 9%

Total Concentration (ppb) Anatoxin-A - LA - LF - LR - LW - LY - RR - YR Nodularin-R

Mean 1.6 (0.49) 1 4.9 (2.5) 5.7 (2.2) 104 (95) 6.3 (2.5) 15 (9.0) 910 (720) 16 (11) 0.10 (~ 0.010)Median 0.16 (< 0.010) 0.24 (0.015) 0.036 (< 0.010) 1.6 (1.1) 0.067 (< 0.010) 0.10 (0.025) 1.1 (0.59) 0.14 (0.032) 0.10 (< 0.010)Minimum < 0.010 < 0.010 < 0.010 < 0.010 < 0.010 < 0.010 < 0.010 < 0.010 < 0.010Maximum 9.5 54 51 2100 56 200 16000 240 0.19

1 Values outside parenthesis are summary statistics for detections only. Values inside parenthesis include non-detections in summary statistics.

Microcystin Congeners

2006 USGS MCR 2006 USGS MCR Cyanotoxin Mixtures Were Commonly Found in Midwest Lakes and Reservoirs in Cyanotoxin Mixtures Were Commonly Found in Midwest Lakes and Reservoirs in

2006 by LC/MS/MS with Up to 9 Toxins Measured in One Sample.2006 by LC/MS/MS with Up to 9 Toxins Measured in One Sample.

0

5

10

15

20

25

1 2 3 4 5 6 7 8 9

Nu

mb

er S

amp

les

Number of Samples with At Least "x" Toxins

Comparison of ELISA Results for MicrocystinsPhotic Zone Sampling vs. Blooms and Accumulations

Higher Detection Frequencies and Higher Concentrations were Observed in Blooms and Accumulations than in Photic Zone Sampling.

Study objectives dictate sample collection procedures (i.e. photic zone vs blooms Study objectives dictate sample collection procedures (i.e. photic zone vs blooms and accumulations, etc.).and accumulations, etc.).

Total Concentration (ppb)

2007 EPA National Lake

Assessment

2006 USGS Midwestern US Reconnaissance

Sample Type Photic Zone, Deepest Blooms and Accummulations

% Detections of Microcystins 32 % (401/1238) 1 100 % (23/23)

Mean 3.0 (~1.0) 2 600 (570)Median 0.52 (< 0.10) 3.0 (2.8)Minimum 0.10 (< 0.10) < 0.10Maximum 230 13000

1 Values in parenthesis are detections divided by total number of samples.

2 Values outside parenthesis are summary statistics for detections only. Values inside parenthesis include non-detections in summary statistics.

Take Home Message…

Cyanotoxins are presentCyanotoxins are present at levels that may be of concern for human health and at levels that may be of concern for human health and ecological impact in some cases. Specifically, microcystins are found nationwide ecological impact in some cases. Specifically, microcystins are found nationwide and are frequently encountered in the Midwest.and are frequently encountered in the Midwest.

Anatoxin-a occurred in 30% of the 2006 USGS Midwest Recon at levels up to 9.5 Anatoxin-a occurred in 30% of the 2006 USGS Midwest Recon at levels up to 9.5 ppb.ppb.

Predictably, Predictably, higher detection frequencies and higher concentrations of toxins higher detection frequencies and higher concentrations of toxins can be can be encountered in cyanobacterial blooms and accumulations versus photic encountered in cyanobacterial blooms and accumulations versus photic zone sampling.zone sampling.

In the 2007 EPA study (photic zone sampling), In the 2007 EPA study (photic zone sampling), 9 (0.7 %) lakes exceeded WHO 9 (0.7 %) lakes exceeded WHO recreational guidelines (20 ppb)recreational guidelines (20 ppb) and 143 (12 %) lakes exceeded WHO drinking and 143 (12 %) lakes exceeded WHO drinking water guidelines (1.0 ppb) for microcystins.water guidelines (1.0 ppb) for microcystins.

In the 2006 USGS Midwest Recon (Bloom and Accumulation sampling), In the 2006 USGS Midwest Recon (Bloom and Accumulation sampling), 4 (17 %) 4 (17 %) lakes exceeded WHO recreational guidelines (20 ppb)lakes exceeded WHO recreational guidelines (20 ppb) and 18 (78 %) lakes and 18 (78 %) lakes exceeded WHO drinking water guidelines (1.0 ppb) for microcystins. Microcystin-exceeded WHO drinking water guidelines (1.0 ppb) for microcystins. Microcystin-LR, one of the most toxic microcystin congeners, was detected most frequently LR, one of the most toxic microcystin congeners, was detected most frequently (91 %).(91 %).

LC/MS/MS results indicate that LC/MS/MS results indicate that toxin mixtures may be fairly commontoxin mixtures may be fairly common..

Jennifer GrahamJennifer Graham jlgraham@usgs.gov jlgraham@usgs.gov

(785) 832-3511(785) 832-3511

Keith LoftinKeith Loftinkloftin@usgs.govkloftin@usgs.gov

(785) 832-3543(785) 832-3543

Additional Information Available on the Web:Additional Information Available on the Web:

Cyanobacteria Cyanobacteria - - http://ks.water.usgs.gov/Kansas/studies/qw/cyanobacteria

USGS cyanotoxin sampling protocol: http://pubs.usgs.gov/sir/2008/5038