title: Effects of environmental regimens on the toxin profile of Alexandrium ostenfeldii.
abstract: Environmental conditions are key factors in the development of marine toxic phytoplankton. Spirolides are marine toxins with a heptacyclic imine ring responsible for the toxicity in mice. Alexandrium ostenfeldii (A. ostenfeldii) is the main producer of these toxins, although this dinoflagellate often produces toxins belonging to the paralytic shellfish poisoning (PSP) group. The present study shows the first evidence that external environmental factors can influence the toxin profile produced by the dinoflagellate A. ostenfeldii. The species investigated is indigenous to the North Atlantic coast, and their cells grew under several environmental parameters. Toxin production was measured by means of liquid chromatography-mass spectrometry (LC-MS) and the chromatograms reflect the presence of two spirolides in all cultures; one in the region m/z 692.5, corresponding to 13-desmethyl spirolide C (13-desMeC) and the other in the region m/z 678.5, which corresponds to 13,19-didesmethyl spirolide C (13,19-didesMeC). The physical parameters studied were salinity, culture media, and photoperiod. The highest amount of toxin per cell was obtained when dinoflagellates grew in F/2 and Walne medium, 28 per thousand salinity, and 24 h of light. However, the highest proportion of 13,19-didesMeC with respect to 13-desMeC was achieved in L1 medium, 33 per thousand salinity, and 14:10 h light:dark. On the contrary, the highest proportion of 13-desMeC in cells was obtained when A. ostenfeldii was cultured in F/2 medium, 28 per thousand salinity, and the same photoperiod. Therefore, from these data the optimum conditions to culture A. ostenfeldii and to obtain the highest amount of spirolide per cell are shown. In addition, these environmental conditions can be considered a tool to predict and avoid A. ostenfeldii blooms.
authors.affiliation: Departamento de Farmacología, Facultad de Veterinaria, Universidad de Santiago de Compostela, 27002 Lugo, Spain.
authors.forename: Paz
authors.initials: P
authors.lastname: Otero
authors.affiliation: Departamento de Farmacología, Facultad de Veterinaria, Universidad de Santiago de Compostela, 27002 Lugo, Spain.
authors.forename: Amparo
authors.initials: A
authors.lastname: Alfonso
authors.affiliation: Departamento de Fisiología, Facultad de Veterinaria, Universidad de Santiago de Compostela, 27002 Lugo, Spain.
authors.forename: Mercedes R
authors.initials: MR
authors.lastname: Vieytes
authors.affiliation: ANFACO-CECOPESCA, Vigo, 36310 Pontevedra, Spain.
authors.forename: Ana G
authors.initials: AG
authors.lastname: Cabado
authors.affiliation: ANFACO-CECOPESCA, Vigo, 36310 Pontevedra, Spain.
authors.forename: Juan M
authors.initials: JM
authors.lastname: Vieites
authors.affiliation: Departamento de Farmacología, Facultad de Veterinaria, Universidad de Santiago de Compostela, 27002 Lugo, Spain.
authors.forename: Luis M
authors.initials: LM
authors.lastname: Botana
mesh_terms: D002851:Chromatography, High Pressure Liquid; D003470:Culture Media; D004141:Dinoflagellida / Q000472:pathogenicity*; D008027:Light; D008387:Marine Toxins / Q000032:analysis*; D013058:Mass Spectrometry; D012965:Sodium Chloride / Q000494:pharmacology
chemical_list: D003470:Culture Media; D008387:Marine Toxins; D012965:Sodium Chloride
references.citation: MacKinnon SL, Cembella AD, Burton IW, Lewis N, LeBlanc P, Walter JA. 2006. Biosynthesis of 13-desmethyl spirolide C by the dinoflagellate Alexandrium ostenfeldii. J Org Chem 71: 8724-8731.
references.citation: MacKinnon SL, Cembella AD, Quilliam MA, LeBlanc P, Lewis NI, Hardstaff WR, Burton IW, Walter JA. 2004. The caracterization of two new spirolides isolated from Danish Strains of the toxigenic dinoflagellate Alexandrium ostenfeldii. In Steidinger KA, Landsberg JH, Tomas CR, Vargo GA, eds, Harmful Algae 2002. Florida Fish and Wildlife Conservation, Commission, Florida Institute of Oceanography, and Intergovernmental Oceanographic Commission of UNESCO, FL, USA. pp 186-188.
references.citation: MacKinnon SL, Walter JA, Quilliam MA, Cembella AD, Leblanc P, Burton IW, Hardstaff WR, Lewis NI. 2006. Spirolides isolated from Danish strains of the toxigenic dinoflagellate Alexandrium ostenfeldii. J Nat Prod 69: 983-987.
references.citation: Sleno L, Windust AJ, Volmer DA. 2004. Structural study of spirolide marine toxins by mass spectrometry. Part I. Fragmentation pathways of 13-desmethyl spirolide C by collision-induced dissociation and infrared multiphoton dissociation mass spectrometry. Anal Bioanal Chem 378: 969-976.
references.citation: Cembella AD, Bauder AG, Lewis NI, Quilliam MA. 2001. Association of the gonyaulacoid dinoflagellate Alexandrium ostenfeldii with spirolide toxins in size-fractionated plankton. J Plankton Res 23: 1413-1419.
references.citation: Gill S, Murphy M, Clausen J, Richard D, Quilliam M, MacKinnon S, LaBlanc P, Mueller R, Pulido O. 2003. Neural injury biomarkers of novel shellfish toxins, spirolides: A pilot study using immunochemical and transcriptional analysis. Neurotoxicology 24: 593-604.
references.citation: Richard D, Arsenault E, Cembella AD, Quilliam M. 2001. Investigations into the toxicology and pharmacology of spirolides, a novel group of shellfish toxins. In Hallegraeff GM, Blackburn SI, Bolch CJ, Lewis RJ, eds, Harmful Algal Blooms 2000. United Nations Educational, Scientific, and Cultural Organization, Paris, France, pp 383-386.
references.citation: Gribble K, Keafer B, Quilliam A, Cembella AD, Kulis D, Manahan A, Anderson D. 2005. Distribution and toxicity of Alexandrium ostenfeldii (Dinophyceae) in the Gulf of Maine, USA. Deep-Sea Res II 52: 2745-2763.
references.citation: Tillmann U, John U, Cembella AD. 2007. On the allelochemical potency of the marine dinoflagellate Alexandrium ostenfeldii against heterotrophic and autotrophic protists. J Plankton Res 29: 527-543.
references.citation: Hu T, Burton IW, Cembella AD, Curtis JM, Quilliam MA, Walter JA, Wright JL. 2001. Characterization of spirolides a, c, and 13-desmethyl c, new marine toxins isolated from toxic plankton and contaminated shellfish. J Nat Prod 64: 308-312.
references.citation: Aasen J, MacKinnon SL, LeBlanc P, Walter JA, Hovgaard P, Aune T, Quilliam MA. 2005. Detection and identification of spirolides in norwegian shellfish and plankton. Chem Res Toxicol 18: 509-515.
references.citation: Ciminiello P, Dell'aversano C, Fattorusso E, Forino M, Grauso L, Tartaglione L, Guerrini F, Pistocchi R. 2007. Spirolide toxin profile of adriatic Alexandrium ostenfeldii cultures and structure elucidation of 27-hydroxy-13,19-didesmethyl spirolide C. J Nat Prod 70: 1878-1883.
references.citation: Alfonso A, Vieytes MR, Botana AM, Goenaga X, Botana LM. 1993. Preparation of mixtures of paralytic shellfish toxin (PSP) standars from mussel hepatopancreas. Fresenius J Anal Chem 345: 212-216.
references.citation: Jensen MO, Moestrup O. 1997. Autecology of the toxic dinoflagellate Alexandrium ostenfeldii: Life history and growth at different temperatures and salinities. Eur J Phycol 32: 9-18.
references.citation: Cembella AD, Bauder AG, Lewis NI, Quilliam MA. 2001. Population dynamics and spirolide composition of the toxigenic dinoflagellate Alexandrium ostenfeldii in coastal embayments of Nova Scotia. In Hallegraeff GM, Blackburn SI, Bolch CJ, Lewis RJ, eds, Harmful Algal Blooms 2000. United Nations Educational, Scientific, and Cultural Organization, Paris, France, pp 173-176.
references.citation: John U, Quilliam MA, Medline L, Cembella AD. 2001. Spirolide production and photoperiod-dependent growth of the marine dinoflagellate Alexandrium ostenfeldii. In Hallegraeff GM, Blackburn SI, Bolch CJ, Lewis RJ, eds, Harmful Algal Blooms 2000. United Nations Educational, Scientific, and Cultural Organization, Paris, France, pp 299-302.
references.citation: Maclean C, Cembella AD, Quilliam MA. 2003. Effects of light, salinity and inorganic nitrogen on cell growth and spirolide production in the marine dinoflagellate Alexandrium ostenfeldii (Pauslsen) Balech et Tangen. Bot Mar 46: 466-476.