title: Amphidoma languida (Amphidomatacea, Dinophyceae) with a novel azaspiracid toxin profile identified as the cause of molluscan contamination at the Atlantic coast of southern Spain.
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 160μg AZA-1 eq. kg-1(reaching maximum levels of >500μg AZA-1 eq. kg-1in Chamelea gallina and >250μg AZA-1 eq. kg-1in 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.
authors.affiliation: Alfred-Wegener-Institut Helmholtz-Zentrum für Polar- und Meeresforschung, Am Handelshafen 12, D-27570 Bremerhaven, Germany. Electronic address: urban.tillmann@awi.de.
authors.forename: Urban
authors.initials: U
authors.lastname: Tillmann
authors.affiliation: Laboratorio de Control de Calidad de los Recursos Pesqueros, Ctra. Punta Umbría-Cartaya, km 12, 21459 Cartaya, Spain.
authors.forename: David
authors.initials: D
authors.lastname: Jaén
authors.affiliation: Laboratorio de Control de Calidad de los Recursos Pesqueros, Ctra. Punta Umbría-Cartaya, km 12, 21459 Cartaya, Spain.
authors.forename: Lourdes
authors.initials: L
authors.lastname: Fernández
authors.affiliation: Department Biologie, Systematische Botanik und Mykologie, GeoBio-Center, Ludwig-Maximilians-Universität München, Menzinger Str. 67, D-80638 München, Germany.
authors.forename: Marc
authors.initials: M
authors.lastname: Gottschling
authors.affiliation: Bruker Daltonik GmbH, Fahrenheitstr. 4, 28359 Bremen, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Witt
authors.affiliation: Centro de Investigacións Mariñas, Apdo 13. 36620 Vilanova de Arousa, Spain.
authors.forename: Juan
authors.initials: J
authors.lastname: Blanco
authors.affiliation: Alfred-Wegener-Institut Helmholtz-Zentrum für Polar- und Meeresforschung, Am Handelshafen 12, D-27570 Bremerhaven, Germany.
authors.forename: Bernd
authors.initials: B
authors.lastname: Krock
mesh_terms: D000818:Animals; D049872:Bivalvia / Q000737:chemistry*; D002853:Chromatography, Liquid; D004141:Dinoflagellida / Q000737:chemistry*; D005517:Foodborne Diseases / Q000469:parasitology; D008387:Marine Toxins / Q000032:analysis / Q000633:toxicity*; D012758:Shellfish / Q000032:analysis*; D013030:Spain; D013141:Spiro Compounds / Q000032:analysis / Q000633:toxicity*; D053719:Tandem Mass Spectrometry; D000095702:Polyether Toxins
chemical_list: D008387:Marine Toxins; D013141:Spiro Compounds; C406592:azaspiracid; D000095702:Polyether Toxins
keywords: AZA-43; Amphidoma; Azaspiracids; New compound; Shellfish contamination; Spain