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Amphidoma languida (Amphidomatacea,Dinophyceae) with a novel azaspiracid toxin profile identified as the cause of molluscan contamination at the Atlantic coast of southern Spain
Affiliation:1. Alfred-Wegener-Institut Helmholtz-Zentrum für Polar- und Meeresforschung, Am Handelshafen 12, D-27570 Bremerhaven, Germany;2. Laboratorio de Control de Calidad de los Recursos Pesqueros, Ctra. Punta Umbría-Cartaya, km 12, 21459 Cartaya, Spain;3. Department Biologie, Systematische Botanik und Mykologie, GeoBio-Center, Ludwig-Maximilians-Universität München, Menzinger Str. 67, D-80638 München, Germany;4. Bruker Daltonik GmbH, Fahrenheitstr. 4, 28359 Bremen, Germany;5. Centro de Investigacións Mariñas, Apdo 13. 36620 Vilanova de Arousa, Spain;1. Bachok Marine Research Station, Institute of Ocean and Earth Sciences, University of Malaya, 16310 Bachok, Kelantan, Malaysia;2. Third Institute of Oceanography, SOA, Xiamen, 361005, China;3. Faculty of Resource Science and Technology, Universiti Malaysia Sarawak, 94300, Kota Samarahan, Sarawak, Malaysia;4. State Key Laboratory in Marine Pollution, Department of Biomedical Sciences, City University of Hong Kong, Hong Kong, 999077, China;5. IFREMER, Department ODE, Laboratory Environment and Resources Bretagne Occidentale, Station de Biologie Marine, Place de la Croix, 29900, Concarneau, France;6. Alfred Wegener Institut-Helmholtz Zentrum für Polar- und Meeresforschung, Am Handelshafen 12, D-27570, Bremerhaven, Germany;1. Tunisian National Agronomic Institute (INAT), U.R Marine Biology (FST El Manar I), IRESA - Carthage University, 43 Avenue Charles Nicolle, 1082 Tunis, Tunisia;2. UMR 9190 MARBEC IRD-Ifremer-CNRS-Université de Montpellier, Place Eugène Bataillon, Case 093, 34095 Montpellier Cedex 5, France;3. Toxin/Natural Products Chemistry Program, National Ocean Service/NOAA, Hollings Marine Laboratory, 331 Fort Johnson Road, Charleston, SC 29412, USA;4. IFREMER, Laboratoire Environnement et Ressources Bretagne Occidentale, Station de Biologie Marine, Place de la Croix, 29900 Concarneau, France;1. Third Institute of Oceanography, Ministry of Natural Resources, Xiamen 361005, China;2. Ifremer, LER BO, Station de Biologie Marine, Place de la Croix, BP40537, F-29185 Concarneau CEDEX, France;3. Alfred Wegener Institute for Polar and Marine Research, Am Handelshafen 12, D-27570 Bremerhaven, Germany;4. School of Earth and Ocean Sciences, University of Victoria, OEASB A405, P. O. Box 1700 16 STN CSC, Victoria, British Columbia, V8W 2Y2, Canada;5. National Marine Environmental Monitoring Center, Ministry of Ecology and Environment, Dalian, 116023, China;6. Cawthron Institute, 98 Halifax Street East, Private Bag 2, Nelson 7042, New Zealand;7. NORCE Climate, NORCE Norwegian Research Centre AS, Bjerknes Centre for Climate Research, Jahnebakken 5, 5007 Bergen, Norway;1. Department of Pharmacology and Therapeutics, University College Cork, Cork, Ireland;2. Environmental Research Institute, University College Cork, Cork, Ireland;3. School of Biological, Earth and Environmental Sciences, University College Cork, Cork, Ireland;4. School of Food and Nutritional Sciences, University College Cork, Cork, Ireland;1. Department of Life Science, College of Natural Sciences, Hanyang University, Seoul 04763, South Korea;2. Alfred Wegener Institute, Helmholtz Center for Polar and Marine Research, Am Handelshafen 12, D-27570 Bremerhaven, Germany;3. Northwest Fisheries Science Center, National Marine Fisheries Service, National Oceanic and Atmospheric Administration, 2725 Montlake Blvd. E., Seattle, WA 98112, USA;4. United States Food and Drug Administration, Center for Food Safety and Applied Nutrition, 5001 Campus Drive, College Park, MD 20740, USA
Abstract:Azaspiracids (AZA) are a group of food poisoning phycotoxins that are known to accumulate in shellfish. They are produced by some species of the planktonic dinophycean taxon Amphidomataceae. Azaspiracids have been first discovered in Ireland but are now reported in shellfish from numerous global sites thus showing a wide distribution. In shellfish samples collected in 2009 near Huelva (Spain), AZA was also found along the Andalusian Atlantic coast for the first time. Analysis using LC–MS/MS revealed the presence of two different AZA analogues in different bivalve shellfish species (Chamelea gallina, Cerastoderma edule, Donax trunculus, and Solen vagina). In a number of samples, AZA levels exceeded the EU regulatory level of 160 μg AZA-1 eq. kg?1 (reaching maximum levels of >500 μg AZA-1 eq. kg?1 in Chamelea gallina and >250 μg AZA-1 eq. kg?1 in Donax trunculus) causing closures of some local shellfish production areas. One dinophyte strain established from the local plankton during the AZA contamination period and determined as Amphidoma languida was in fact toxigenic, and its AZA profile disclosed it as the causative species: it contained AZA-2 as the main compound and the new compound AZA-43 initially detected in the shellfish. AZA-43 had the same mass as AZA-3, but produced different collision induced dissociation (CID) spectra. High resolution mass spectrometric measurements indicated that there is an unsaturation in the H, I ring system of AZA-43 distinguishing it from the classical AZA such as AZA-1, -2, and -3. Furthermore, the Spanish strain was different from the previously reported AZA profile of the species that consist of AZA-38 and AZ-39. In molecular phylogenetics, the Andalusian strain formed a monophyletic group together with other strains of Am. languida, but ITS sequences data revealed surprisingly high intragenomic variability. The first Andalusian case of AZA contamination of shellfish above the EU regulatory limit reported here clearly revealed the risk of azaspiracid poisoning (AZP) for this area and also for the Atlantic coast of Iberia and North Africa. The present study underlines the need for continuous monitoring of AZA and the organisms producing such toxins.
Keywords:Azaspiracids  New compound  AZA-43  Shellfish contamination  Spain
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