1. Research and Other Activities Related To Harmful Algal Blooms (HABs)
United States Environmental Protection Agency
Office of Water and Office of Research and Development
Research and Other Activities Related To
Harmful Algal Blooms (HABs)
February 9, 2016
Christopher Impellitteri (ORD) and Lesley D’Anglada (OW)

2. Disclaimer
The views expressed in this presentation are those of the authors and do not necessarily represent the views or policies of the U.S. Environmental Protection Agency.

3. Analytical methods development - April 2015
Regulations, Guidelines and Recommendations
Cyanobacteria and their toxins included in the Contaminant Candidate List (CCL)
CCL 1, 2, 3, draft 4
Analytical methods development - April 2015
544 (microcystins and nodularin-R)
545 (anatoxin-a and cylindrospermopsin)
Drinking Water Health Advisories (10-day) - June 2015
Microcystins (0.3 µg/L for < 6yr old, 1.6 µg/L for 6 yrs-adult)
Cylindrospermopsin (0.7 µg/L for < 6yr old, 3.0 µg/L for 6 yrs-adult)
Recommendations for Public Water Systems to Manage Cyanotoxins in Drinking Water - June 2015

4. Drinking Water Protection Act (H.R. 212)
Regulations, Guidelines and Recommendations
Drinking Water Protection Act (H.R. 212)
Signed on August 7th, 2015 to amend the SDWA by adding Section 1459
Directs EPA to develop and submit a strategic plan for assessing and managing risks associated with algal toxins in drinking water provided by public water systems
Includes steps and timelines to assess:
Human health effects, list of algal toxins, health advisories, treatment options, analytical and monitoring approaches, causes of HABs, source water protection, and collaboration and outreach.
Identifies information gaps and publishes information from federal agencies studying algal toxins.

5. Regulations, Guidelines and Recommendations
H.R. 212 Strategic Plan
“Algal Toxin Risk Assessment and Management Strategic Plan for Drinking Water”
Developed with input from:
EPA Regional offices, ORD, and OW
Federal partners, including the IWG-HABHRCA
Stakeholders through a listening session webinar
Submitted to Congress November 5, 2015

6. Recreational Ambient Water Quality Criteria (AWQC) for cyanotoxins
Regulations, Guidelines and Recommendations
Recreational Ambient Water Quality Criteria (AWQC) for cyanotoxins
Clean Water Act  §304(a) recommended recreational water quality criteria values protective of human health given a primary contact recreational exposure scenario.
OW is developing AWQC recommendations for microcystins and cylindrospermopsin.
Expected Date: Draft Fall 2016

7. Assessment of Cyanotoxins
Unregulated Contaminant Monitoring Rule (UCMR) 4
Monitoring of cyanotoxins in drinking water public systems from 2018 to 2020.
National Aquatic Resource Surveys (NARS)
Studies of the nation's aquatic resources designed to report on the condition of lakes, rivers/streams, coastal waters and wetlands.
Survey parameters: Indicators associated with the risk of potential exposure to cyanotoxins.
National Lakes Assessment (2007, 2012)
Microcystins, cyanobacteria and chlorophyll-a
National Wetland Condition Assessment (2011, 2016)
Microcystins
National Rivers and Stream Assessments ( )
Microcystins and chlorophyll-a
National Coastal Condition Assessment (2015)
Microcystins and chlorophyll-a, and additional algal toxins

8. Outreach and Communications
EPA’s Cyanobacteria Harmful Algal Blooms Webpage
Freshwater HABs Newsletter
Fact Sheets
Cyanobacteria and Cyanotoxins: Information for Drinking Water Systems
Climate Change and Harmful Algal Blooms
Stakeholder Engagement through webinars and public meetings
Recreational Ambient Water Quality Criteria for Cyanotoxins
Recommendations for public water systems to manage cyanotoxins in drinking water
Inland HABs Discussion Group
EPA Regional Workshops on HABs ( )
Provide technical support to States and tribal agencies working on HABs-related issues in fresh and coastal waters and provide opportunities for collaborations with national and regional partners

9. Partnerships and Collaborations
IWG-HABHRCA
National HABs Committee (NHC)
Cyanobacteria Assessment Network (CyAN) Project
Great Lakes HABs Collaboratory
Source Water Collaborative
Great Lakes Interagency Task Force
Other Partnerships and Collaborations (e.g. Association of Clean Water Administrators (ACWA), Association of State Drinking Water Administrators (ASDWA), Ground Water Protection Council (GWPC))
EPA Regions Partnerships through MOA, Grants and Cooperative Agreements
Region 1 Cyanobacteria Monitoring & Bloom Watch Program
Harmful Algal Blooms Observing System (HABSOS) Binational Workshops on HABs and Environmental Measurements

10. ORD’s HABs-Related Research

11. SSWR HAB Research Areas
Water Quality
Human and Ecological Health Effects
Monitoring/ Analytical Methods
Drinking Water Treatment
The Safe and Sustainable Water Resources program has a large effort on harmful algal blooms. To help with managing the projects, they are broken up into four categories
1) Water Quality,
2) Human and Ecological Health Effects,
3) Monitoring/Analytical Methods,
4) Drinking Water Treatment.
Today, I’ll give you a rundown of the projects in each area, with a few project highlights. On the phone, we have the researchers to answer any questions you may have. Due to the limited time and the large number of studies, perhaps we can save the questions to the end.

12. Water Quality Research
Watershed/Source water HAB risk mitigation
HAB related water quality trading research
In the area of Water Quality studies, there are two main areas of work:
Source Waters-Impacts from HABs on Disinfection By-Product Formation
(POCs-Joel Allen, Chris Nietch, Alan Lindquist, Ed Barth, and Michael Elovitz (NRMRL))
- Model reservoir algal community dynamics and link to DW treatment and recreational uses.
HAB related water quality trading (Matt Heberling, Hale Thurston, Chris Nietch-NRMRL).
- Inclusion of treatment costs due to HABs in water quality trading framework.

13. East Fork Watershed and Lake Harsha as a case study
Study Example:
Water Quality Trading (WQT)
Goal: Improve WQT programs nationwide
Cost-effectively reduce nutrient loadings
Estimating cost savings to participants associated with reduced nutrient loading in streams
Transferable to other watersheds
East Fork Watershed and Lake Harsha as a case study
Drinking Water Intake
Determine if non-traditional participants have an incentive to purchase credits:
DWTPs to protect source water
Recreationists to protect beaches
Local homeowners to protect nearby streams
To improve water quality trading programs nationwide, our interdisciplinary research will determine if non-traditional participants such as drinking water treatment plants to protect source water, recreationists to protect beaches, and local homeowners to protect nearby streams, have an incentive to purchase credits too. Using the East Fork Watershed and Harsha Lake in southwest Ohio as a case study, we are estimating the cost savings to potential participants associated with reduced nutrient loading in streams. The monitoring, modeling, and economic approaches are transferable to other watersheds.
Nutrient Trading
Excess agricultural nutrient runoff is the major cause of impaired waters in the USA and regulatory authority to manage this source is not included in the Clean Water Act.
Water quality trading offers a voluntary market driven means of reducing nutrient loadings and delisting of impaired waters.
Attractive potential solution from environmental, economic and political perspectives.
Insufficient buyers and/or sellers are the chief reasons WQ markets fail.
We are evaluating the potential to establish a WQ trading market (nitrogen) in the watershed drained by the East Fork of the Little Miami River (HUC12).
Farmers are the sellers
Potential buyers include the drinking water treatment plant, recreational users of Lake Harsha, State swimming competition, rowing competition
Realization of a functional trading market in the EFLMR will require quantification, bundling and marketing of the various final ecosystem goods (need a term more meaningful to the audience here)  and services provided by the watershed.
Nitrogen
Phosphorus
HAB mitigation
Greenhouse gas mitigation
Stream temperature control
Etc.
Preliminary data from 2014 hyperspectral flyover:
Multi-agency ground-truthing study for HAB remote sensing algorithms.

14. Human & Ecological Health Effects Research
Characterization of environmental strains of cyanobacteria and their corresponding toxic and allergenic components
Toxic gene-specific monitoring for harmful algal bloom using meta- transcriptomic and RT-qPCR approaches
A data intensive investigation of temperature impacts and bloom modelling
Adverse Mammalian Health Effects of Algal Toxins: Bioaccumulation, Bloom Extract Toxicity, and Basic Mechanisms of Action
For the area of Human and Ecological Health Effects studies, there are a number of projects. They are listed here in no particular order of importance:
Characterization of Environmental Strains of Cyanobacteria and Their Corresponding Toxic and Allergenic Components (POCs-Armah de la Cruz, Dennis Lye, Jerry Stelma-NERL, Jorge Santo Domingo (NRMRL))
- Purify and characterize endotoxins from cyanobacteria.
Perform in vitro assays to determine allergenicity of endotoxins.
Toxic Gene-Specific Monitoring for Harmful Algal Bloom using Meta-transcriptomic and RT-qPCR Approaches POCs: Jingrang Lu and Jim Lazorchak (NERL)
Develop qPCR assays for source cyanobacteria in source water
Monitor upregulated genes using transcriptomic analysis
A Data Intensive Investigation of Temperature Impacts and Bloom Modelling (POCs-Betty Kreakie and Jeff Hollister-NHEERL).
- Data acquisition and database development.
- Temperature modelling.
- Bloom modelling.
- Future scenarios.
Adverse Mammalian Health Effects of Algal Toxins: Bioaccumulation, Bloom Extract Toxicity, and Basic Mechanisms of Action (POC-Neil Chernoff-NHEERL).
- Mechanistic study of cyanotoxin toxicity
- Compare toxicity of bloom extracts with toxicity of mixtures of pure toxins
Gene expression alteration following cyanotoxin exposure
The Effects of Selected Cyanotoxins on Human Cells and Cell Lines
Effects of multiple congeners of microcystins on cells (colonocytes, hepatocytes, renal cells)
Human Exposure to and Health Effects from Cyanotoxins (POC-Elizabeth Hilborn-NHEERL)
- Develop methods to confirm biologic evidence of human and animal exposure to cyanotoxins.
- Use population-based epidemiology and existing health databases to characterize microcystin health effects.
HAB associated disease outbreaks among users of freshwater lakes in the United States.
Understanding Harmful Algal Blooms and Protecting the Ecosystem and Human Health from their Toxins (POCs-Jim Lazorchak, John Darling-NERL; Joel Allen, Chris Nietch-NRMRL)
- Examine impacts from algal toxins on aquatic life.
- Comparison of real-time biomonitoring systems with traditional toxicity tests.
- Identification of conditions that promote golden algae blooms.
The Effects of Selected Cyanotoxins on Human Cells and Cell Lines
Human exposure to, and health effects from, cyanotoxins
Protecting the ecosystem and human health from HAB toxins

15. Characterize Strains of Cyanobacteria
Study Example:
Characterize Strains of Cyanobacteria
Cells
Development of improved method to isolate cyanobacteria and determine toxigenic properties
Identification of cyanobacteria and their toxins
Purify and characterize endotoxins from cyanobacteria
In vitro assays to determine allergenicity of endotoxins
The outcomes of this study will provide researchers with expertise in (1) the identification of cyanobacteria and their toxins, (2) the isolation and culturing of cyanobacteria from the environment, (3) the purification and characterization of LPS and (4) the performance of the in vitro -hexosaminidase release assay for allergens. In addition, a well-characterized collection of cultures from the environment will be obtained for future studies. The development of improved method to isolate cyanobacteria and determine their toxigenic properties is part of EPA-ORD’s Safe and Sustainable Water Resources (SSWR) Research Action Plan Task 2.2B.
Microcystis
Microcystin
Toxin

16. Monitoring/Analytical Methods Research
Satellite Cyanobacteria Assessment Network
Rapid cyanobacteria cell detection
High frequency monitoring of HABs to understand interactions between watershed nutrients and drinking water safety
Tribal-Focused Exposure and Risk Screening Tool (T-FERST) through predictive forecasting of natural toxin blooms
In the area of Monitoring/Analytical Methods, there are also a number of projects:
Satellite Cyanobacteria Assessment Network (POC-Blake Schaeffer-NERL).
- Detect and quantify cyanobacteria blooms in the US using satellite data records.
Rapid Cyanobacteria Cell Detection (POC-Robert Zucker-NHEERL).
- Develop methods to quickly and accurately identify different cyanobacteria.
High Frequency Monitoring of Harmful Algal Blooms: Understanding Interactions Between Watershed Nutrients and Drinking Water Safety (POCs-Joel Allen, Michael Elovitz, Chris Nietch, Jake Beaulieu-NRMRL; Jim Lazorchak-NERL)
- Real-time monitoring for verification of USACE remote sensing algorithms.
- Development of empirical relationships among algal communities, cyanotoxins, climate, and water quality parameters.
Tribal-Focused Exposure and Risk Screening Tool (T-FERST) through Predictive Forecasting of Natural Toxin Blooms (NTBs)
POCs: Tammy Jones-Lepp and Dan Heggem (NERL)
Develop monitoring and forecasting tools for HAB events in Tribal watersheds.
Analytical Method Development:
- Optimization of Method 544-microcystins and nodularin in drinking water using LC/MS/MS. (POC-Jody Shoemaker-NERL).
- Optimization of Methods 544 and 545 (for cylindrospermopsin and anatoxin-a) for ambient water samples (POC-Jody Shoemaker-NERL).
- Identification and verification of analytical methods to measure microcystin concentrations in source waters (POCs-Heath Mash, Darren Lytle, Nicholas Dugan-NRMRL).
Compare/contrast microcystin analytical results from ELISA and LC/MS/MS measurements.
Science to Achieve Results (STAR) Research projects (POC-Angela Page-NCEA)
- Near-real Time, Highly Sensitive and Selective Field Deployable Biosensor for Cyanotoxins and Cyanobacteria Using both Antibodies and DNA-signatures.
- Development and application of a fiber optic array system for detection and enumeration of potentially toxic cyanobacteria.
Development of analytical methods for determination of microcystin and nodularin concentrations
Science to Achieve Results (STAR) HAB-related research

17. Development of Analytical Methods
Study Example:
Development of Analytical Methods
Detection of cyanobacterial toxins
Optimization of Method 544:
Detection of microcystins and nodularin in drinking water by SPE & LC/MS/MS
Optimization of Methods 544 and 545L:
Detection of cylindrospermopsin and anatoxin-a in ambient water by SPE & LC/MS/MS
Comparison of ELISA and LC/MS/MS:
Compare results of ELISA measurements and LC/MS/MS concentrations of microcystin variants in source water
Optimization of Method 544-microcystins and nodularin in drinking water using LC/MS/MS. (POC-Jody Shoemaker-NERL).
Optimization of Methods 544 and 545 (for cylindrospermopsin and anatoxin-a) for ambient water samples (POC-Jody Shoemaker-NERL).
Identification and verification of analytical methods to measure microcystin concentrations in source waters (POCs-Heath Mash, Darren Lytle, Nicholas Dugan-NRMRL). Compare/contrast microcystin analytical results from ELISA and LC/MS/MS measurements.

18. Drinking Water Treatment Research
Toledo Ohio Pilot-Scale Water Treatment Facility for the Bloom Season
Evaluating the Impact of Algicides on a Cyanobacterial Cell’s Propensity to Release Toxins during the Early Stages of the Drinking Water Treatment Process
In the area of drinking water treatment studies, there are two projects.
Toledo Ohio Pilot-Scale Water Treatment Facility for the 2016 Bloom Season (POC-Darren Lytle-NRMRL)
- Installation of pilot plant in Toledo to evaluate treatment strategies for Lake Erie
Evaluating the Impact of Algicides on a Cyanobacterial Cell’s Propensity to Release Toxins during the Early Stages of the Drinking Water Treatment Process
(POCs-Nicholas Dugan, Darren Lytle, Heath Mash, Toby Sanan-NRMRL)
- Effects of copper sulfate and hydrogen peroxide-based algicides on the release of cyanotoxins.

19. Lake Erie DW Treatment Plant Studies
Study Example:
Lake Erie DW Treatment Plant Studies
Field Studies
Monitoring cyanobacteria toxins through numerous treatment plants on Lake Erie
Fate/transport of toxins through treatment systems and treatment effectiveness
Lake Erie
Bench- and Pilot-Scale Studies
Bench-scale permanganate, ozone, and activated carbon trials
Pilot installation (Toledo) anticipated in time to be operational for the 2016 bloom
Lake Erie Studies The sampling campaign provided a unique opportunity to characterize the development of Lake Erie’s cyanobacterial bloom and its associated toxins at a high level of analytical detail. Since different classes of toxins, and even variants within the same class, exhibit differing degrees of treatability, tracking their possible propagation or removal through a treatment facility allows utilities and regulators to make better-informed long term decisions regarding the operation and modification of treatment processes to optimize removals.
Here are the plants along Lake Erie. Only a subset of them are tested each year.
Bench- and pilot-scale Studies
Divided into two phases: a series of bench-scale studies performed over the winter of 2014/2015, and pilot-scale field studies performed at partner drinking water treatment plants during the 2015 bloom season. Will improve existing treatment systems by modifying the locations where chemicals are applied, the types and concentrations of chemicals applied, and the pH at which the processes are operated.
Bench-scale studies – will be performed with Lake Erie raw water spiked with microcystins, and will be subdivided into ozone and non-ozone trials. The ozone trials will focus on the impacts of pH and hydrogen-peroxide addition on ozone contactor efficiency. The results of the bench-scale tests will inform the design of the pilot-scale studies.
Pilot systems – will be installed in time to be fully operational for the 2015 bloom season and will be monitored for the control of cyanobacterial toxins, while ensuring that the effluents continue to comply with federal standards for turbidity and disinfection by-product formation.
Will support technical guidance to regions, states, municipalities

20. Contact Information
Office of Water
Lesley V. D’Anglada
Senior Microbiologist
OW/Office of Science and Technology
Office of Research and Development
Christopher Impellitteri
Associate National Program Director
Safe and Sustainable Water Resources

21. Questions? 12