USING MOLECULAR METHODS TO DETERMINE THE ROLE OF PHYTOPLANKTON IN WEST COAST SHELLFISH DIE-OFFS

The community members of Lummi Nation are the main stakeholders for this research, and they have identified shellfish die-offs as a problem. Community members have requested that we implement research tools available at the SSRC to mitigate closures and shellfish die-offs.We will use molecular methods to identify and quantify phytoplankton, called dinoflagellates, that produce yessotoxins (YTX).We aim to use a suite of molecular techniques to identify toxic YTX- producing algae in marine waters and shellfish and determine the concentration of YTX in samples from the Salish Sea, WA and locations in California.This proposed research would enhance our capacity to use genomic tools to monitor for a devastating shellfish killer and support tribal interests in safe seafood harvests.?Goal 1: Develop species-specific quantitative PCR (qPCR) Taqman assays to identify the following algae species that produce yessotoxins: (1) L. polyedrum, (2) P. reticulatum, and (3) G. spinifera, (YTX-producing algae).Objective 1: In-situ testing of the Taqman assay using samples collected from sites with L. polyedrum, P. reticulatum, and G. spinifera presence from surface water collections during the regular bloom periods in the Salish Sea and coastal California.Objective 2: In-situ testing of the Taqman assay with collection of shellfish from sites with suspected presence of P. reticulatum, and G. spinifera.Goal 2: Compare qPCR data with the following: (1) relative abundance of phytoplankton data for toxin species, (2) SPATT analyses for YTX by LC-MS/MS, (3) corresponding environmental data.Objective 1: Determine if qPCR results for concentration of L. polyedrum, P. reticulatum, and G. spinifera from surface water correlate to microscopy data that details the relative concentration of L. polyedrum, P. reticulatum, and G. spinifera.Objective 2: Compare qPCR results for concentration of L. polyedrum, P. reticulatum, and G. spinifera with YTX biotoxin concentrations from SPATT data.Objective 3: Investigate the presence of YTX-producing phytoplankton with nutrient and environmental data in order to better understand the causes of shell-fish die-offs.Goal 3: Increase capacity of students at NWIC and Lummi community members to conduct genomics-related research projects to help tribal communities manage tribal resources.Objective 1: Engage students in summer genomic research projects that are support tribal priorities of safe shellfish harvests.Objective 2: Offer an eDNA workshop.Goal 4: Broaden the investigation of YTX-related shellfish deaths from the Salish Sea to sampling locations in California.Objective 1: Provide mentorship of undergraduate SSRC interns by graduate students from UCSC.Objective 2: Examine both local and regional environmental patterns and broadly study YTX related shellfish deaths on the west Coast of the United States.