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1. Application of Six Detection Methods for Analysis of Paralytic Shellfish Toxins in Shellfish from Four Regions within Latin America

Author

Andrew D. Turner, Alejandra B. Goya, Frances M. Van Dolah, Maria Salhi, Benjamin A. Suarez-Isla, Robert G. Hatfield, Maggie Broadwater, Dinorah Medina, Fernanda Barrera, Mickael Teixeira-Alves, Ignacio Rubilar, D. Carrasco, Sophie Tarnovius, Ernesto García-Mendoza, Turner A. D., Tarnovius S., Hatfield R. G., Alves M. T., Broadwater M., Van Dolah F., Garcia-Mendoza E., Medina Dinorah, DINARA, Salhi María, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Ecología y Ciencias Ambientales. DINARA, Goya A. B., Barrera F., Carrasco D., Rubilar I., and Suarez-Isla B. A.

Subjects
0106 biological sciences, toxin profiles, Latin Americans, MBA, Pharmaceutical Science, Biology, RBA, 01 natural sciences, Article, Mice, Mouse bioassay, Paralytic shellfish poisoning (PSP), Tandem Mass Spectrometry, Toxicity Tests, Drug Discovery, Lc ms ms, medicine, Animals, Paralysis, Shellfish Poisoning, False Positive Reactions, paralytic shellfish poisoning (PSP), LC-MS/MS, Paralytic shellfish poisoning, lcsh:QH301-705.5, Pharmacology, Toxicology and Pharmaceutics (miscellaneous), Chromatography, High Pressure Liquid, Shellfish, Replacement method, business.industry, 010604 marine biology & hydrobiology, 010401 analytical chemistry, Reproducibility of Results, Reference Standards, Toxin profiles, medicine.disease, Bivalvia, 0104 chemical sciences, Shellfish poisoning, Biotechnology, Latin America, lcsh:Biology (General), Biological Assay, Marine Toxins, LC-FLD, business
Abstract

With the move away from use of mouse bioassay (MBA) to test bivalve mollusc shellfish for paralytic shellfish poisoning (PSP) toxins, countries around the world are having to adopt non-animal-based alternatives that fulfil ethical and legal requirements. Various assays have been developed which have been subjected to single-laboratory and multi-laboratory validation studies, gaining acceptance as official methods of analysis and approval for use in some countries as official control testing methods. The majority of validation studies conducted to date do not, however, incorporate shellfish species sourced from Latin America. Consequently, this study sought to investigate the performance of five alternative PSP testing methods together with the MBA, comparing the PSP toxin data generated both qualitatively and quantitatively. The methods included a receptor binding assay (RBA), two liquid chromatography with fluorescence detection (LC-FLD) methods including both pre-column and post-column oxidation, liquid chromatography with tandem mass spectrometry (LC-MS/MS) and a commercial lateral flow assay (LFA) from Scotia. A total of three hundred and forty-nine shellfish samples from Argentina, Mexico, Chile and Uruguay were assessed. For the majority of samples, qualitative results compared well between methods. Good statistical correlations were demonstrated between the majority of quantitative results, with a notably excellent correlation between the current EU reference method using pre-column oxidation LC-FLD and LC-MS/MS. The LFA showed great potential for qualitative determination of PSP toxins, although the findings of high numbers of false-positive results and two false negatives highlighted that some caution is still needed when interpreting results. This study demonstrated that effective replacement methods are available for countries that no longer wish to use the MBA, but highlighted the importance of comparing toxin data from the replacement method using local shellfish species of concern before implementing new methods in official control testing programs.

Published
2020
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2. Evaluation of Rapid, Early Warning Approaches to Track Shellfish Toxins Associated with Dinophysis and Alexandrium Blooms.

Author

Hattenrath-Lehmann TK, Lusty MW, Wallace RB, Haynes B, Wang Z, Broadwater M, Deeds JR, Morton SL, Hastback W, Porter L, Chytalo K, and Gobler CJ

Subjects
Animals, Chromatography, Liquid methods, Environmental Monitoring methods, Mytilus edulis parasitology, Seafood analysis, Seafood parasitology, Seawater parasitology, Tandem Mass Spectrometry methods, Dinoflagellida chemistry, Marine Toxins chemistry, Shellfish analysis, Shellfish parasitology
Abstract

Marine biotoxin-contaminated seafood has caused thousands of poisonings worldwide this century. Given these threats, there is an increasing need for improved technologies that can be easily integrated into coastal monitoring programs. This study evaluates approaches for monitoring toxins associated with recurrent toxin-producing Alexandrium and Dinophysis blooms on Long Island, NY, USA, which cause paralytic and diarrhetic shellfish poisoning (PSP and DSP), respectively. Within contrasting locations, the dynamics of pelagic Alexandrium and Dinophysis cell densities, toxins in plankton, and toxins in deployed blue mussels ( Mytilus edulis ) were compared with passive solid-phase adsorption toxin tracking (SPATT) samplers filled with two types of resin, HP20 and XAD-2. Multiple species of wild shellfish were also collected during Dinophysis blooms and used to compare toxin content using two different extraction techniques (single dispersive and double exhaustive) and two different toxin analysis assays (liquid chromatography/mass spectrometry and the protein phosphatase inhibition assay (PP2A)) for the measurement of DSP toxins. DSP toxins measured in the HP20 resin were significantly correlated ( R ² = 0.7-0.9, p < 0.001) with total DSP toxins in shellfish, but were detected more than three weeks prior to detection in deployed mussels. Both resins adsorbed measurable levels of PSP toxins, but neither quantitatively tracked Alexandrium cell densities, toxicity in plankton or toxins in shellfish. DSP extraction and toxin analysis methods did not differ significantly ( p > 0.05), were highly correlated ( R ² = 0.98-0.99; p < 0.001) and provided complete recovery of DSP toxins from standard reference materials. Blue mussels ( Mytilus edulis ) and ribbed mussels ( Geukensia demissa ) were found to accumulate DSP toxins above federal and international standards (160 ng g -1 ) during Dinophysis blooms while Eastern oysters ( Crassostrea virginica ) and soft shell clams ( Mya arenaria ) did not. This study demonstrated that SPATT samplers using HP20 resin coupled with PP2A technology could be used to provide early warning of DSP, but not PSP, events for shellfish management., Competing Interests: The authors declare no conflict of interest.

Published
2018
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