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MERHAB Fiscal Year 2004 Projects

California Program for Regional Enhanced Monitoring of PhycoToxins
(Cal-PReEMPT)

Institution(s): University of California Santa Cruz (UCSC), California Department of Health Services (CDHS)

Investigator(s): Peter E. Miller, Raphael Kudela, Mary W. Silver (UCSC) and

Gregg W. Langlois (CDHS)

Partners: Center for Integrated Marine Technology, Center for Integrative Coastal Observation, Research and Education, Monterey Bay National Marine Sanctuary, Quileute Tribe, La Push, Washington, Central Coast Long-term Environmental Assessment Network (CCLEAN)

Abstract:

The goal of this MERHAB effort is to implement an economically sustainable harmful algal bloom monitoring plan for the California coastline that exceeds current capabilities of the California Department of Health Services (CDHS) by using new technologies for rapid toxin and species detection and bloom tracking. The innovative approach taken establishes pilot project sites where new technologies are incorporated into an intensive monitoring program, in combination with a tiered decision-making protocol that dictates specific steps to take in response to field observations. The power of this approach is that it paves the way for ultimately shifting much of the monitoring effort to the field, where a network of volunteers, with overall guidance from the CDHS, pre-screen samples using new technologies, thus ensuring early warning of impending blooms while avoiding unnecessary and expensive lab-based sample testing. Using the best available remote sensing data in conjunction with field data provided by the volunteer force will enable tracking the inception, proliferation, advection and decline of bloom events in real-time along the expansive California coast. In turn, this provides managers necessary information to make informed decisions on when and where to direct the field force to increase their efforts.

A particular strength of this program is that it was jointly conceived and developed between the state agency charged with ensuring seafood safety, the CDHS, and academic researchers actively engaged in developing and using new methods for harmful algal bloom research. This effort represents a true collaborative effort that specifically addresses the needs of the CDHS with regard to maintaining a high level of safety, while at the same time recognizing the limitations imposed by economic realities.

In Situ Nutrient Monitoring and Eutrophication-Related Blooms

Institution: University of Maryland Center for Environmental Science Horn Point Laboratory

Investigator(s): Patricia M. Glibert Louis A. Codispoti, University of Maryland Center for Environmental Science Horn Point Laboratory

Abstract:

The goal of this proposal is to continue development of methodologies and approaches that lead to efficient and cost-effective monitoring of nutrients and eutrophication- related blooms. In the Chesapeake and Coastal Bays of Maryland a range of harmful species occur in response to nutrient inputs. This project builds on investment of the past several years in in situ nutrient monitoring equipment and development. The next step is to couple this capability with fine-scale collection of water samples for organismal detection to resolve time-dependent relationships which ultimately can be incorporated into models. Remote water sampling will be triggered by pre-set changes in nutrient concentration and salinity for analysis of HAB species and other chemical parameters. A new sensor for urea will also be developed, as this form of nitrogen is abundant in Chesapeake Bay tributaries. Improved telemetry of nutrient data will also be developed to allow the posting of nutrient data in real time. Results to date have demonstrated that traditional approaches for collecting information on nutrient distributions - such as weekly or bimonthly sampling - are inadequate to resolve the short-lived nutrient pulses that follow meteorological events. Increases in P043, N03 and N}T occur following major freshwater flow events, and these increases are ephemeral. The response of chlorophyll to these nutrient inputs occurs on a short-term scale. What is not known is the response by different species within the phytoplankton community, and whether different HAB species have differential responses. This information is necessary if predictive models by species are to be developed. Results will continue to be integrated with results from other monitoring efforts by Maryland State agencies, as has been done in the past. Results will also prove useful to managers in other regions plagued by HABs and eutrophication. By developing a system to rapidly post the temperature, salinity, chlorophyll, and nutrient data collected autonomously, managers will be provided with relevant water quality data in real time.

Quantitative Molecular Detection of Multiple HAB Species

Institution: University of Delaware, Lewes, DE

Investigator(s): Craig Cary and Kathryn Coyne, University of Delaware

The project will develop quantitative real-time polymerase chain reaction (QRT-PCR) methodology for low-cost, high-throughput detection and enumeration of multiple HAB species. This methodology will include efficient, cost-effective and user-friendly protocols for sample collection, nucleic acid extraction, and QRT-PCR analysis of environmental water samples. In addition, the methodology will be flexible in design to accommodate regionally-specific groups of HAB species. This methodology will be transferred to state monitoring agencies through a series of workshops that will include hands-on training for participants. Working closely with monitoring personnel, the investigators will also prepare a detailed manual of protocols developed in this project that can be integrated into existing HAB monitoring programs.

The project will fully investigate methods for environmental sample collection and extraction of high quality DNA for QRT-PCR analysis. These protocols will be optimized specifically to increase the efficiency of sample preparation. The investigators will develop methodology for the multi-species QRT-PCR detection in a single-tube format using primers and probes to target HAB species indigenous to coastal waters along both the Atlantic and Pacific coasts of the US. Detection sensitivity and quantitative range will be optimized for each multi-species platform using environmental samples that have been spiked with known concentrations of each target species. The technologies developed in this investigation will be transferred to state and Federal monitoring personnel through a series of training workshops and the dissemination of a manual of protocols specifically for the collection, extraction and QRT­PCR analysis of targeted HAB species from environmental samples. The manual will be supported by a web site with species updates and suggestions for improvements.

Monitoring Domoic Acid in Marine Food Webs and Water

Institution(s): NOAA Northwest Fisheries Science Center; Quileute Natural Resources, and NOAA Center for Coastal Fisheries and Habitat Restoration

Investigator(s): Vera Trainer, NOAA Northwest Fisheries Science Center; Jay Burns, Quileute Natural Resources; Pat Tester and Wayne Litaker, NOAA Center for Coastal Fisheries and Habitat Restoration

The project will develop sensitive cost effective detection methods for domoic acid (DA), the toxin produced by diatoms of the genus Pseudo-nitzchia, DA that can be performed in tribal laboratory facilities. The specific objectives of this study are as follows:

  1. To assess whether the RAPID DA assay designed to detect closure levels of domoic acid are reliable. This involves extracting clams and crabs with a 50% methanol solution and comparing the results of the RAPID DA assay with HPLC analyses don on a subset of the same samples;
  2. To complete development and evaluation of an existing colorometric competition enzyme-linked immunosorbent assay (ELISA) capable of detecting sublethal, as well as lethal, levels of domoic acid in clams and crabs.
  3. Adapt the colorometric ELISA to an electrochemical format that can be carried out in the field.
  4. Use both ELISA methods to examine how DA concentrates in the food web, particularly leading to DA accumulation in crabs; determine which environmental conditions are consistently associated with high levels of DA in the water column; examine how free DA in the water column is related to bloom activities and to what extent free DA can serve as an early warning system for toxic events; and to estimate sub lethal exposure levels associated with routine consumption of clams and crabs.
  5. To transfer the technology to other tribes and state health officials.

MERHAB Fiscal Year 2002 Projects

Alexandrium Bloom Transport: Observation and Models

Institution: Woods Hole Oceanographic Institution

Principal Investigators: D.J. McGillicuddy, D.M. Anderson, B.A. Keafer

The overall objective of this project is to obtain field data on A. fundyense  cell distributions and local hydrodynamics and to interpret those data using existing numerical models and data products from GoMOOS to determine if they can be used to provide short-term forecasts of bloom transport. Specific tasks are to: 1) collect field data in the western Gulf of Maine at temporal and spatial scales suitable for resolving A. fundyense distributions within the area covered by a CODAR array; 2) use CODAR and Acoustic Doppler Current Profiler (ADCP) measurements to estimate surface water velocities; 3) assimilate velocity observations into an existing physical/biological coupled model; 4) seed the hindcast simulation with passive particles to assess transport of A. fundyense; 5) compare observed and predicted A. fundyense distributions with PSP toxicity records; and  6) assess future needs for transitioning these predictive tools to meet management needs.

http://www.whoi.edu/science/AOPE/people/olga/mm_main_page.html

MERHAB Lower Great Lakes

Institutions: Research Foundation for the State University of New York (SUNY); SUNY Brockport; University at Buffalo; University of Vermont; Western Michigan University; New York State Sea Grant; University of Tennessee.

Investigators: Gregory L. Boyer, SUNY-College of Environmental Science and Forestry; Joseph Makarewicz, SUNY Brockport; Joe Atkinson University at Buffalo; Mary Watzin, University of Vermont; Tim Mihuc, Western Michigan University; Charles O'Neil New York State Sea Grant; Steven Wilhelm University of Tennessee.

Related Websites:
Toxin Lab Established by CSCOR Helps Ohio Agencies Respond to Toxic Freshwater Algal Blooms
MERHAB-LGL at ESF
State University of New York at Brockport - MERHAB Research
MERHAB-Lower Great Lakes Program, State University of New York at Buffalo
MERHAB-Lower Great Lakes Program, University of Tennessee
MERHAB-Lower Great Lakes Program, Western Michigan University
Rubenstein Ecosystem Science Laboratory at the Lake Champlain Science Center

Abstract:

This proposal will develop an integrated alert system to monitor and detect toxic cyanobacteria blooms in the lower Great Lakes: Lake Erie, Lake Ontario and Lake Champlain. The proposal is organized around six different working groups, each with their own tasks. The Lake Erie working group will investigate the spatial distribution of toxic Microcystis in Lake Erie, evaluate the chemical diversity of microcystin(s) produced in the lake, evaluate the use of molecular markers for the microcystin biosynthesis genes mcyB and mcvD as monitoring tools for toxigenic species, and examine nutritional probes for iron, nitrogen and phosphorus as predictors for toxic cyanobacterial blooms. The Champlain working group will investigate the occurrence of anatoxin-a and microcystins in Lake Champlain, including the identification of the phytoplankton species responsible for toxin formation in this system, examine the correlation between blue-green algal density and toxin production, validate a newly developed dipstick assay for anatoxin-a, evaluate cyanotoxin screening protocols for potential use by water treatment operators, and develop training programs for those water quality managers. The Lake Ontario group will examine the occurrence of toxic cyanobacteria in the Lake Ontario's southern shore embayments and determine if these embayments are a source of cyanobacteria and toxins to the open lake water and to the St Lawrence river. It will also examine the potential of using zebra mussels as surrogate monitoring system (mussel watch). A centralized toxin support group will analyze for the cyanobacteria toxins including microcystins, anatoxin-a, anatoxin-a(s), PSP toxins and cylindrospermopsin. They will also develop extraction methodology and form an HAB event response team. A remote sensing working group will provide information on the occurrence/movement of phytoplankton blooms in the region and apply new remote sensing platforms to the occurrence of toxic cyanobacteria blooms. Finally an education working group will disseminate this material to concern parties by developing a public awareness program for cyanobacteria toxins, informing and educate local environmental, health, and monitoring agencies integrating the groups field studies into information on management strategies. detection techniques, health risks, and what is likely to be an appropriate public response.

Eastern Gulf of Mexico Sentinel Program

Institution: Florida Fish & Wildlife Conservation Commission-Florida Marine Research Institute

Investigator: Brian Bendis, Florida Fish and Wildlife Commission

Abstract:

Blooms of the red-tide dinoflagellate, Karenia brevis, occur annually in Florida coastal waters of the eastern Gulf of Mexico (GOMx) and present serious impacts to marine resources, public health, and community economics. To minimize these impacts, early forecasting of and subsequent mitigation for bloom events are the goal of federal, state, academic, and private partnerships. The difficulty in forecasting occurrence and impacts of K brevis blooms can be traced to the absence of appropriate monitoring technologies. Clearly, the successful, operational utility of any monitoring program (and any resulting mitigation strategies) will be dependent upon synoptic-scale, real-time sampling, regional-based data assimilation and modeling and adaptive, event-response confirmation.

The Eastern GOMx Sentinel Program is a multi-investigator and multi- disciplinary program to develop and assess the utility of a networked system of autonomous sampling platforms incorporating physical/chemical- and bio-sensor packages.. As such, it will facilitate model and forecast initializations and state-wide, adaptive field sampling. Monitoring platforms for sensor deployment will utilize both existing and newly established buoys (The West Florida Coastal Ocean Monitoring and Prediction System) and Bottom-Stationed Ocean Pro filers (autonomous, water-column profiling vehicles carrying modular sensor payloads). The bio-sensors, including a bio-optical phytoplankton discriminator and a processor containing a molecular-probe array, are existing first generation instruments that previously have provided positive, consistent results in both laboratory and field trials and show great promise for remote, autonomous discrimination of K. brevis. Generated data will be used to initialize a coupled bio- physical model for forecasting the development and movement of K. brevis red tides. In addition, the data can be used for (near) real-time 'ground-truthing' of a NOAA/NOAA Ocean Service forecast tool based on satellite ocean-color imagery and other forecast tools.

Platform-based data acquisition and forecasting efforts will be integrated with a geographically-comprehensive, rapid response component that incorporates adaptive, field sampling and a state-sanctioned, volunteer sampling network. Vessel- and shore-based, adaptive sampling (and subsequent analyses) will verify offshore- and near-shore blooms, respectively. The volunteer monitoring program, in existence since May, 2000 and extending from Key West to Pensacola, FL, will be supplemented with a shore-based volunteer network to provide additional coverage.

The Eastern GOMx Sentinel Program is intended to become a strategic, proactive hardware- and information-technology for operational coastal observatories within the GOMx. The program is designed for technological integration and data dissemination into existing (and future) federal, state, academic and private partnerships with their associated data management and communication network. NOAA's National Coastal Data Development Center, a vital figure in the Northern GOMx demonstration project, HABSOS, will receive, store and layer the hydrological and biological data for dissemination and future archiving. Products arising from this program will help fulfill scientific directives of the Alliance for Coastal Technologies, particularly the demonstration, evaluation, and verification of monitoring sensors, platforms, and software.

Websites for more information:
   http://research.myfwc.com/
   http://ocgweb.marine.usf.edu
   http://comps.marine.usf.edu
   http://coolgate.mote.org/socool/

In Situ Optical Early Warning System to Detect Harmful Algal Blooms

Institution: Texas A&M University

Investigator: Dr. Lisa Campbell (TAMU/OCN)

Abstract:

The objective of this research is to develop a buoy-based in situ continuous monitoring system capable of detecting increases in abundance of specific cell types that could form the basis for a real-time early warning system for harmful algae blooms (HABS). Occurrences of HABs have increased at alarming rates over the past 20-30 years. In the Gulf of Mexico, the toxic dinoflagellate Karenia brevis (formerly known as Gymnodinium breve) is a major hannful algal bloom species. Toxic algal bloom events along the Texas coast have occurred in five of the past six years and present serious health and economic risks. Early detection of potential blooms and a rapid response to such events have been suggested as the most effective ways to mitigate the impact of HABs. In some states with recurring blooms, such as Texas, there is no regular monitoring program in place for early warning. Plans for development of the US Coastal Ocean Observing System often mention detection of HABs as an important objective, which requires automated continuous monitoring with real-time data access. The proposed targeted research program is directed towards this goal. The scientific objective of this project is to test a novel optical detection system, the FlowCAM (Flow Imaging Technologies, Inc.) in conjunction with the existing Texas Automated Buoy System (TABS) and modeling program as a new tool for early warning of HAB events. Following laboratory and field tests to validate automatic counting and to optimize data retrieval, the FlowCAM together with an in situ nutrient analyzer and oxygen sensor will be installed on a new buoy located off the coast at Corpus Christi, the site of a number of recent algal blooms. This submersible system will analyze, image and count cells in continuous flow. A digital camera will capture and record images of individual cells for both immediate transmittal back to shore via cell phone and for archival. K. brevis has a characteristic shape and should be easily recognized by the optics system. Results will provide improved efficiency of image capture and transmittal of data, which are essential for early detection. Data will be linked directly to the existing TABS web site for real-time display of phytoplankton images. TABS also runs, on a daily basis, numerical models that nowcast and forecast currents along the Texas coast. Because these capabilities are already operational and TABS scientists and technicians have years of experience in offshore buoy operations, work can concentrate specifically on the technology of measuring and forecasting harmful algal blooms. From continuous measurements of cell abundance, together with nutrient and oxygen concentrations, temperature, salinity and currents, changes in phytoplankton community structure and abundances of K brevis and other potentially harmful species can be determined on a scale never before possible. This program will contribute to understanding the effects of nutrient loading on HABs. Texas A&M University is committed to undergraduate and graduate education. This project will provide opportunity and training for student(s) in coastal ocean observing, an oceanographic specialty that will provide an increasing number of jobs for graduates over the next decade.

Immunoassays for Florida Red Tide Monitoring

Institution: University of North Carolina, Wilmington

Investigator: Jerome Naar, Center for Marine Science, UNCW

Abstract:

Blooms of toxic or harmful micro algae, COmIn,only called red tides, represent a significant and expanding threat in the US with an estimated annual loss of $49 Million by Gulf industries (1). In the Gulf of Mexico, red tides are caused by blooms of the toxic dinoflagellate Karenia brevis (ex Gymnodinium breve). These blooms result in ecological disasters such as fish, marine mammal and sea bird deaths as well as contaminated seafood and human respiratory distress along shorelines.

During the last year, we completed development of a new enzyme immunoassay (EIA) for brevetoxin analysis in complex matrices such as seawater, shellfish and mammalian body fluid (Naar et al. 2002). The assay is designed for research, industry and regulatory use. The advantages of this particular assay are: 1) Sensitivity: detection limit for brevetoxins is in the nanomolar range, 2) Specificity: Only brevetoxins and brevetoxin metabolites are measured, 3) Simplicity: Seawater, shellfish and fish tissue, bird and mammalian body fluids, and sea-aerosol can be analyzed without extraction of the samples. Regarding shellfish monitoring, the ELISA method is more rapid than any other analysis because it can be performed on shellfish homogenate without preliminary extraction and/or purification. This means that the total amount of toxin is measured, regardless of whether it is sequestered in the meat or free in interstitial water in the homogenate, and whether or not it is lipid- soluble. This assay is 40 times more sensitive than the mouse bioassay, and within 8 hours, a single person can analyze over 50 shellfish samples. Thus with automated gear, and the abbreviated preparation scheme, shellfish samples can be measured daily to permit more rapid re-opening of beds, individual shellfish variability can be measured, and a variety of depuration experiments can be carried out. ELISA technology does not require radioactive tracers, live animals, solvents, extraction equipment, or expensive materials and facilities, thus it appears to be very useful for monitoring brevetoxins in both biological and environmental samples.

The objective of the work described hereafter is to incorporate the ELISA technology into the actual monitoring program for brevetoxins in the Gulf States. To reach this objective, water and shellfish samples will be analyzed using 1) the current monitoring protocol (cell counts and mouse bioassays respectively) and 2) ELISA technology. Furthermore as a confmnatory technique, selected samples will be also analyzed by mass spectrometry. The tandem analyses will provide:

1. Correlation between cell counts and the actual amount of toxin present in the water.

2. Comparison between the toxicity of oysters as assessed by the mouse bioassy with the concentration of total toxin present in the shellfish as measured by enzyme immunoassay (EIA).

3. Determination of the actual concentration of brevetoxins present in shellfish using current re- opening criteria (20 MU toxicity). This value will serve as the basis in redefining perniissible levels of brevetoxins in shellfish as measured by EIA.

Development and Field-Testing of an Analytical Protocol for Pfiesteria Toxin

Institution: Old Dominion University

Investigator: Andrew Gordon, Old Dominion University

Abstract:

Dinoflagellates within the toxic Pfiesteria complex (TPC) can cause extensive fish kills and produce toxin(s) that are implicated in serious human heath effects. The currently accepted diagnosis for a Pfiesteria-mediated fish kill requires sequential testing including (1) demonstration of the presence of300 or more Pfiesteria-like organisms (PLO) ml-1 ; (2) demonstration of the presence of toxic Pfiesteria by standardized laboratory fish bioassay; (3) confirmation of species identification by electron microscopy of suture-swollen cells; and (4) corroboration of identification by additional laboratories. Additionally molecular detection methods (PCR and real-time PCR) are now in use by a number of laboratories.

Although this approach is the best available diagnostic procedure, it is confounded by the fact that not all TPC organisms produce toxin(s). Even Pfiesteria strains that produce toxins don't always produce them. Thus even if isolates from water from a fish kill are shown to be Pfiesteria spp and to have the potential to produce ichthyotoxin in a laboratory bioassay, it does not prove that they were toxic in situ.

A more direct and rapid approach to diagnosis of a Pfiesteria-mediated fish kill would be demonstration of Pfiesteria toxin or its products at toxic levels in the water at the kill site. This, in addition to positive molecular or direct count evidence of high PLO numbers, would be sufficient to implicate TPC's as a causative agent. However an analytical protocol for Pfiesteria toxin and/or its products is not presently available.

We have been cultivating toxic Pfiesteria spp. for four years and have recently completed analyses of stability of toxic activity in cell-free filtrates. Preliminary results we have obtained utilizing solid phase separation to concentrate toxic activity from water samples indicate quantitative removal is readily achievable and that toxic products can be eluted in a form suitable for analysis by high performance liquid chromatography (HPLC). We are in the relatively unique position of having access to toxic Pfiesteria cultures in our own laboratory and to the expertise and equipment required for development of an analytical protocol for the toxic products of these organisms.

In this proposal funds are requested to complete development and field-testing of an HPLC-based analytical procedure for Pfiesteria toxin(s) and/or their toxic products. This will yield a rapid, powerful, and definitive tool to enhance our ability to determine if Pfiesteria caused a fish kill and whether potential human health threats exist at a bloom site. The analysis will use instrumentation and methods that are widely available and relatively inexpensive so, once optimized, it will be applicable by most laboratories involved in HAB research and monitoring. Although extensive Pfiesteria-mediated fish kills have not occurred recently, they will undoubtedly occur in the future and a field tested analytical protocol for their toxic products will be an essential addition to the "tool kit" available to researchers and regulators.

Detecting Karenia brevis Blooms in the Western Gulf of Mexico

Institution: University of Texas at Austin

Investigator: Tracy Villareal, The University of Texas at Austin

Abstract:

The proposed work will expand the existing satellite-based monitoring program for the Gulf of Mexico to include the Texas coast (western Gulf). From 1935 to 1986, blooms of the toxic, dinoflagellate, Karenia brevis, affected the region intermittently. Since then, the frequency of events has increased dramatically with over half the documented red tides occurring in the last decade. Unlike Florida, Texas has no large-scale monitoring program, nor is one likely to be developed in the near future due to funding pressures within the State. The state agencies charged with recording fish mortality and closing shellfish beds respond to fish kills or fortuitous observations as tripwire indicators. Thus, implementing satellite and modeling capabilities for routine remote detection and monitoring is the only practical means for covering the state's extensive offshore area. NOAA's existing satellite-based harmful algal bloom program in Florida has been successful in detecting blooms. The resulting data sets have been incorporated into a distribution product, the NOAA Harmful Algal Bloom (HAB) Bulletins, that identifies HAB locations and short-term movements, and directing event response to them. This is an invaluable tool for agencies responsible for regulating shellfish harvesting and providing information for recreational and commercial fishers.

The proposed work will modify this application for the Texas coast and couple it to a focused 3-year monitoring program for model and algorithm verification. Water color characteristics and current patterns are unique to the Texas coast and require site-specific sampling for validation. A simple cross-shelf wind-driven circulation model will be initialized with satellite data and available field data to predict the shot1-term (days) initiation and movement of the bloom. The field program uses a collaboration with Texas Parks and Wildlife Department to provide offshore (9 nautical miles) samples at no cost to this program. These predictions will be tested using event response sampling (cell counts, chlorophyll) based out of the Marine Science Institute (UT -Austin).

The outcome of this project will be a near-real-time tool for detecting and predicting K. brevis events along the Texas coast. The product will be integrated into the NOAA HAB bulletins and the proposed Harmful Algal Bloom Observing System (HABSOS) program in order to provide a near-real time, web-accessible, HAB visualization product.

Tier-based Monitoring for Toxic Cyanobacteria in the Lower Great Lakes

Institution: Research Foundation for the State University of New York (SUNY)

Investigator: Gregory L. Boyer, State University of New York

Abstract:

This proposal will develop an integrated alert system to monitor and detect toxic cyanobacteria blooms in the lower Great Lakes: Lake Erie, Lake Ontario and Lake Champlain. The proposal is organized around six different working groups, each with their own tasks. The Lake Erie working group will investigate the spatial distribution of toxic Microcystis in Lake Erie, evaluate the chemical diversity of microcystin(s) produced in the lake, evaluate the use of molecular markers for the microcystin biosynthesis genes mcyB and mcvD as monitoring tools for toxigenic species, and examine nutritional probes for iron, nitrogen and phosphorus as predictors for toxic cyanobacterial blooms. The Champlain working group will investigate the occurrence of anatoxin-a and microcystins in Lake Champlain, including the identification of the phytoplankton species responsible for toxin formation in this system, examine the correlation between blue-green algal density and toxin production, validate a newly developed dipstick assay for anatoxin-a, evaluate cyanotoxin screening protocols for potential use by water treatment operators, and develop training programs for those water quality managers. The Lake Ontario group will examine the occurrence of toxic cyanobacteria in the Lake Ontario's southern shore embayments and determine if these embayments are a source of cyanobacteria and toxins to the open lake water and to the St Lawrence river. It will also examine the potential of using zebra mussels as surrogate monitoring system (mussel watch). A centralized toxin support group will analyze for the cyanobacteria toxins including microcystins, anatoxin-a, anatoxin-a(s), PSP toxins and cylindrospermopsin. They will also develop extraction methodology and form an HAB event response team. A remote sensing working group will provide information on the occurrence/movement of phytoplankton blooms in the region and apply new remote sensing platforms to the occurrence of toxic cyanobacteria blooms. Finally an education working group will disseminate this material to concern parties by developing a public awareness program for cyanobacteria toxins, informing and educate local environmental, health, and monitoring agencies integrating the groups field studies into information on management strategies. detection techniques, health risks, and what is likely to be an appropriate public response.