Temporal and spatial distribution of Azadinium species in the inland and coastal waters of the Pacific northwest in 2014-2018.

Azaspiracids, produced by some species of the dinoflagellate genera Azadinium and Amphidoma, can cause a syndrome in humans called azaspiracid shellfish poisoning (AZP). In 1995, mussels from the Irish west coast contaminated with azaspiracids were, for the first time, linked to this human illness that has symptoms of nausea, vomiting, severe diarrhea, and stomach cramps. The only confirmed cases of AZP to date in the United States occurred in Washington State in 2008 from mussels imported from Ireland. Shortly after this case, several others involving similar gastrointestinal symptoms were reported by shellfish consumers from Washington State. However, no detectable diarrhetic shellfish toxins or Vibrio contamination were found. Cursory analysis of Solid Phase Adsorption Toxin Tracking (SPATT) samplers suggested the presence of azaspiracids in Washington State waters and motivated a study to evaluate the presence and distribution of Azadinium species in the region. During the spring and summer months of 2014-2015, quantitative polymerase chain reaction (qPCR) analyses detected the presence of the toxigenic species Azadinium poporum and A. spinosum on the outer coast and throughout the inland waters of Washington State. In 2016-2018, standard curves developed using A. poporum isolated from Puget Sound and A. spinosum isolated from the North Sea were used to quantify abundances of up to 10,525 cells L of A. poporum and 156 cells L of A. spinosum at shore-based sites. Abundances up to 1,206 cells L of A. poporum and 30 cells L of A. spinosum were measured in the coastal waters of the Pacific Northwest in 2017. Other harmful genera, including Alexandrium, Dinophysis, and Pseudo-nitzschia, were observed using light microscopy at coastal sites where A. poporum was also observed. In some samples where both A. poporum and A. spinosum were absent, an Amphidomataceae-specific qPCR assay indicated that other species of Azadinium or Amphidoma were present. The identification of Azadinium species in the PNW demonstrates the need to assess their toxicity and to incorporate their routine detection in monitoring programs to aid resource managers in mitigating risks to azaspiracid shellfish poisoning in this region.