Institutions: University of South Florida, Florida Atlantic University, Caribbean Coastal Ocean Observing System, US Virgin Islands Department of Planning and Natural Resources, NOAA Atlantic Oceanographic and Meteorological Laboratory

Project Summary: Coastal communities in the southeastern US and Caribbean have been plagued by increasingly severe inundations of floating macroalgae (Sargassum sp.). These inundation or beaching events are considered harmful algal blooms and can have devastating impacts on coastal ecosystems and economies. This project will improve existing Sargassum detection and forecasting technologies; facilitate a better understanding of biochemical impacts associated with inundation or beaching; and foster the autonomous delivery of timely delivery of accurate forecasts and warning. The project team will produce delineated, daily Sargassum distribution maps at coarse-to-fine spatial scales. Forecast trajectories will be computed for each Sargassum patch identified and estimates of biochemical impacts will be determined when inundation is predicted or observed. Improved forecast and warnings will be autonomously integrated across several distribution platforms, allowing users free access to these data passively (e.g., web distribution) or actively through customized alerts. The project directly involves managers and engages an advisory group to help guide the research product development. Improved alerts will give communities in Florida, Puerto Rico, the US Virgin Islands and the wider Caribbean more time to mobilize responses and minimize the harm caused by Sargassum inundation events.

Institutions: Woods Hole Oceanographic Institution, Florida Fish and Wildlife Commission, Bigelow Laboratory for Ocean Sciences, and Northeastern Regional Association of Coastal Ocean Observing Systems (NERACOOS)

Project Summary: Highly productive shellfisheries in the northeast US continue to be challenged by toxic blooms of certain species of the marine diatom Pseudo-nitzschia (PN) which first emerged as a management concern in 2012. Certain PN species can produce domoic acid, a potent neurotoxin. Domoic acid can accumulate in shellfish that feed on PN, and can lead to amnesic shellfish poisoning (ASP) in people who eat tainted shellfish. Regional shellfish agencies have adopted monitoring protocols to maintain seafood safety but a lack of advanced warning can lead to costly, precautionary closures of large harvest areas. This project will identify the underlying drivers of ASP emergence on the Maine coast. The project will track toxic PN blooms as they develop offshore and track bloom progress as they move across US-Canada maritime boundary and toward Maine coastal shellfish beds. The team will track blooms with Imaging FlowCytobot\autonomous surface vessel (IFCB\ASV) mobile platforms. Data from

IFCB-ASV systems will be telemetered to shore by satellite and made publicly accessible alongside remote sensing products and numerical model simulations of bloom transport. Detection of significant blooms entering the Gulf of Maine will trigger charter vessel-based, event driven intensive sampling cruises to determine toxicity and species composition. The project will also establish co-deployments of IFCB and Environmental Sample Processor (ESP) sensors at three stations along the Maine coast to expand an existing HAB observation network for New England (HABON-NE), including two in Downeast Maine and one in Western Maine.

These shore stations will track the evolution and toxicity of PN as blooms are transported west into New Hampshire and Massachusetts waters. All sources of HAB monitoring information will be made available in real-time through an open-source, interactive data access and visualization portal called HABhub. All HABhub data layers, including new remote sensing and numerical model visualizations, will be built to ingest standard data formats, facilitating adoption of HABhub data sharing by other US regions.

Institutions: Gloucester Marine Genomics Institute, Florida Fish and Wildlife Research Institute, Bigelow Laboratory for Ocean Sciences

Project Summary: Toxin production can vary across strains of the same algal species. For example, more than species of the marine diatom Pseudo-nitzschia have been identified, but only certain species of Pseudo-nitzschia produce domoic acid, a potent neurotoxin. Having cost effective and accurate methods to determine if toxin-producing Pseudo-nitzschia species are present is a critical need for agencies working to mitigate bloom impacts. Domoic acid can accumulate in shellfish prompting human health and marine ecosystem concerns for fishery managers in the Gulf of Maine and Florida. This targeted project will enhance existing

Pseudo-nitzschia monitoring efforts with molecular, genetic-based assays. The team will evaluate existing diagnostic tests including a Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-based (SHERLOCK) assay and qPCR-based assays to identify optimal methods for accurate, rapid, cost-effective, sensitive, and portable molecular detection of toxigenic species and in-field estimates of HAB risks by managers and community scientists.

Collaborations with Maine Department of Marine Resources, the New Hampshire Department of Environmental Services, the Massachusetts Division of Marine Fisheries, and Florida’s Department of Agriculture and Consumer Services and Fish and Wildlife Conservation Commission will allow the team to test new methods with field samples, tailor methods to fit

end-user needs, and speed integration of improved approaches into state HAB monitoring and response programs. The project will create and maintain a field sample database and a curated online sample library. It will also tie-in to existing Maine Environmental DNA (eDNA) project and IFCB user networks to promote data sharing, develop protocols and create a communications toolkit in partnership with NERACOOS.

Institutions: San Francisco Estuary Institute, U.S. Geological Survey California Water Science Center, California Department of Water Resources, University of California Santa Cruz, Central Valley Regional Water Quality Control Board, San Francisco Regional Water Quality Control Board, Bend Genetics, LLC.

Project Summary: The goal of this project is to improve monitoring and response to harmful blooms (of both marine and freshwater species) and a suite of algal toxins they produce which have been documented throughout the San Francisco Estuary (SFE), spanning from the upper estuary to the San Francisco Bay. The project team will develop a cost-effective robust

system-wide HAB monitoring program, identify optimal sites for routine monitoring and foster improved coordination among SFE management entities. This project will advance remote sensing applications for HAB detection and response, molecular DNA-based toxin detection, and broaden community HAB monitoring efforts. An Estuary HAB dashboard will integrate all existing and new data facilitating improved understanding and better decision-making. Key knowledge gaps about the transport of freshwater and marine HAB cells (multiple taxa) and toxins (microcystin, saxitoxin, domoic acid) will be addressed by the team using dock and boat-based water grabs, passive in water sampling devices, shellfish and molecular tools. An advisory group with representatives from the San Francisco Water Board, Bay Area Clean Water Association, Baykeeper, State Water Board, CA Dept. of Fish and Wildlife, NOAA Fisheries, Central Valley Water Board, CA Dept. of Water Resources, Delta Stewardship Council, and Restore the Delta will help facilitate transition of research findings to application and build sustainable support for the piloted system-wide HAB monitoring program.

Project Summary: Subsistence harvesting of seafood is a major focus of food resources and cultural practices among the Native villages in Alaska’s Kodiak Archipelago, but food security is threatened by recurrent blooms of toxic Alexandrium and Pseudo-nitzschia species. Similarly, these blooms are a major obstacle to the developing Kodiak mariculture industry. This targeted project will develop increased capacity for monitoring harmful algal blooms and their toxins in Kodiak coastal waters. Monitoring locations will be established at traditional harvesting sites along the Kodiak Road system and data will be shared to help mitigate HAB impacts on rural and Native communities in the City of Kodiak and greater Chiniak Bay area including those of the Sun’aq Tribe of Kodiak, the Native Village of Afognak and the Tangirnaq Native Village, and a significant number of Filipino-Americans. Coordination and information sharing between communities will be strengthened by building on established Alaska Sea Grant (ASG) and the Kodiak Area Native Association (KANA) efforts and with local education and training opportunities. The project will establish a laboratory in Kodiak providing capacity to test shellfish and seawater samples for algal toxins. It will also advance the development of lab-based quantitative molecular methods such as quantitative polymerase chain reaction (qPCR) testing for more rapid, accurate detection of Alexandrium cells and cysts. Capabilities for running lab-based enzyme-linked immunosorbent assay (ELISA) tests will enable better detection of saxitoxins. The team will continue efforts to map Alexandrium cyst distributions in local waters to identify where future blooms may occur and support efforts to develop and validate an Alexandrium HAB forecast model for Kodiak, Alaska. The team will leverage partnerships with the Knik Tribe of Alaska and with a private company developing improved algal toxin tests.

Project Summary: In the lower Chesapeake Bay region, annual blooms of two harmful algal species, Margalefidinium polykrikoides and Alexandrium monilatum, have been linked to shellfish and fish kills, eutrophication and regional oxygen depletion (hypoxia/anoxia). These blooms disrupt oyster aquaculture and bay restoration, degrade coastal ecosystem quality, and can impact local tourism and recreation. This project will pilot a regional monitoring and response network that incorporates a diverse suite of measurements to expand our understanding of the geographic and temporal distribution of HAB species in the region. This information will help inform predictive models that will enable the delivery of HAB forecasts to inform decisions by shellfish growers, state and regional managers, and other stakeholders.

Engagement and training of scientists, growers and citizens on the use of PlanktoScope, a high-resolution digital microscope designed for professional and citizen scientists, will add new phytoplankton image data to existing HAB monitoring efforts. Improved spatial and temporal monitoring data will enable the team to validate remote sensing and in situ sensor observations and support efforts to develop and refine HAB now-cast and forecasts for the region. Data and forecasts will be accessible via the Chesapeake Bay Environmental Forecasting System (CBEFS) and Mid-Atlantic Regional Association Coastal Ocean Observing System portals. This effort will also help advance NCCOS operational HAB forecasting and observing goals for the region.

Institutions: State University of New York College of Environmental Science and Forestry, National Research Council Canada

Project Summary: Cyanobacteria naturally produce toxins. The most commonly found cyanotoxins found in the U.S. are microcystins, cylindrospermopsin, anatoxins and saxitoxins. Many states have robust monitoring programs which are strongly focused on microcystins, a potent liver toxin. While these are widespread, increasingly other species and their toxins are being reported in the Great Lakes. For example, Anabaena and Aphanizomenon are cyanobacteria known to produce anatoxins. Variants of anatoxin can cause respiratory paralysis and have been associated with dog, livestock, and wildlife fatalities. A lack of robust detection methods has resulted in a lack of monitoring programs focused on anatoxins. Currently, only one anatoxin variant is measured using the approved U.S. Environmental Protection Agency (EPA) method 545; anatoxin-a (ATX). This can lead to under-estimates of the total cyanotoxicity in a freshwater body and complicate the accurate assessment of cyanoHAB risks to humans and wildlife. This project will develop peer-validated, certified reference materials and standards for several anatoxin variants including anatoxin-a (ATX), homoanatoxin-a (HTX), and their derivatives. The team project will communicate all results to state agencies, produce a published peer-reviewed method validation helping ensure compatibility across different laboratories, and will share the new method with the EPA to encourage upgrading method 545 to include new anatoxin variants. A partnership with the National Research Council Canada will enable the team to provide standards for general distribution in Canada and abroad.