Azaspiracid Shellfish Poisoning: A Review on the Chemistry, Ecology, and Toxicology with an Emphasis on Human Health Impacts

Reguera, B, Blanco, J, Fernandez, ML, and Wyatt, T (1998). Harmful Algae, Santiago de Compostela.

Satake, 1998, Azaspiracid, a new marine toxin having unique spiro ring assemlies, isolated from Irish mussels, Mytilus edulis, J Amer Chem Soc, 120, 9967, 10.1021/ja981413r

(2006). Risk Assessment of Azaspiracids (AZAs) in Shellfish, August 2006 - A Report of the Scientific Committee of the Food Safety Authority of Ireland (FSAI).

Taleb, 2006, First detection of azaspiracids in mussels in north west Africa, J Shellfish Res, 25, 1067, 10.2983/0730-8000(2006)25[1067:FDOAIM]2.0.CO;2

Nicolaou, 2004, Total synthesis and structural elucidation of azaspiracid-1. Final assigment and total synthesis of the correct structure of azaspiracid-1, J Am Chem Soc, 128, 2859, 10.1021/ja054750q

Botana, LM (2007). Phycotoxins Chemistry and Biochemistry, Blackwell Publishing.

James, 2004, Azaspiracid poisoning, the food-borne illness associated with shellfish consumption, Food Addit Contam, 21, 879, 10.1080/02652030400002105

Nicolaou, 2003, Total synthesis of the proposed azaspiracid-1 structure, Part 2: Coupling of the C1–C20, C21–C27, and C28–C40 fragments and completion of the synthesis, Angew Chem, 42, 3649, 10.1002/anie.200351826

Nicolaou, 2003, Total synthesis of the proposed azaspiracid-1 structure, Part 1: Construction of the enantiomerically pure C1–C20, C21–C27, and C28–C40 fragments, Angew Chem, 42, 3643, 10.1002/anie.200351825

Nicolaou, 2004, Structural revision and total synthesis of azaspiracid-1, Part 2: Definition of the ABCD domain and total synthesis, Angew Chem Int Ed, 43, 4318, 10.1002/anie.200460696

Nicolaou, 2004, Structural revision and total synthesis of azaspiracid-1, Part 1: Intelligence gathering and tentative proposal, Angew Chem, 116, 2

Botana, LM (2007). Phycotoxins chemistry and biochemistry, Blackwell Publishing.

Ofuji, 2001, Structures of azaspiracid analogs, azaspiracid-4 and azaspiracid-5, causative toxins of azaspiracid poisoning in Europe, Biosci Biotechnol Biochem, 65, 740, 10.1271/bbb.65.740

Ofuji, 1999, Two analogs of azaspiracid isolated from mussels, Mytilus edulis, involved in human intoxication in Ireland, Nat Toxins, 7, 99, 10.1002/(SICI)1522-7189(199905/06)7:3<99::AID-NT46>3.0.CO;2-L

Brombacher, 2002, Studies on azaspiracid biotoxins. II. Mass spectral behaviour and structural elucidation of azaspiracid analogs, Rapid Comm Mass Spectrom, 16, 2306, 10.1002/rcm.863

James, 2003, Detection of five new hydroxyl analogues of azaspiracids in shellfish using multiple tandem mass spectrometry, Toxicon, 41, 277, 10.1016/S0041-0101(02)00288-X

Rehmann, 2008, Discovery of new analogs of the marine biotoxin azaspiracid (AZA) in blue mussels (Mytilus edulis) by ultra-performance-liquid-chromatography (UPLC) tandem-mass-spectrometry (MSMS), Rapid Comm Mass Spectrom, 22, 549, 10.1002/rcm.3385

Furey, 2003, Elucidation of the fragmentation pathways of azaspiracids, using electrospray ionisation, hydrogen/deuterium exchange, and multiple-stage mass spectrometry, J Mass Spectrom, 38, 1178, 10.1002/jms.526

Wang, 2004, LC/MS analysis of brevetoxin metabolites in the Eastern oyster (Crassostrea virginica), Toxicon, 43, 455, 10.1016/j.toxicon.2004.02.017

Wilkins, 2006, Identification of fatty acid esters of pectenotoxin-2 seco acid in blue mussels (Mytilus edulis) from Ireland, J Agricult Food Chem, 54, 5672, 10.1021/jf060396j

Yasumoto, 1985, Diarrhetic shellfish toxins, Tetrahedron, 41, 1019, 10.1016/S0040-4020(01)96469-5

Aasen, 2006, Discovery of fatty acid ester metabolites of spirolide toxins in mussels from Norway using liquid chromatography/tandem mass spectrometry, Rapid Comm Mass Spectrom, 20, 1531, 10.1002/rcm.2501

Ciminiello, 2000, Structure determination of carboxyhomoyessotoxin, a new yessotoxin analogue isolated from Adriatic mussels, Chem Res Toxicol, 13, 770, 10.1021/tx000048q

Finch, 2005, Isolation and identification of (44-R,S)-44,55,dihydroxyyessotoxin from Protoceratium reticulatum, and its occurrence in extracts of shellfish from New Zealand, Norway and Canada, Toxicon, 46, 160, 10.1016/j.toxicon.2005.03.007

Satake, 1997, Two new analogs of yessotoxin, homoyessotoxin and 4,5-hydroxyhomoyessotoxin, isolated from mussels of the Adriatic Sea, Nat Toxins, 5, 107, 10.1002/nt.4

Hess, 2007, Fit-for-purpose shellfish reference materials for phycotoxins in internal and external quality control, Anal Bioanal Chem, 387, 2463, 10.1007/s00216-006-0792-8

McCarron, 2007, Feasibility of gamma irradiation as a stabilisation technique in the preparation of tissue reference materials for a range of shellfish toxins, Anal Bioanal Chem, 387, 2487, 10.1007/s00216-006-0935-y

Twiner, 2007, Extraction and analysis of lipophilic brevetoxins (PbTx) from the red tide dinoflagellate Karenia brevis, Anal Biochem, 369, 128, 10.1016/j.ab.2007.06.031

Henshilwood, K, Deegan, B, McMahon, T, Cusack, C, Keaveney, S, Silke, J, O’Cinneide, M, Lyons, D, and Hess, P (2006). Proceedings of the 5th International Conference on Molluscan Shellfish Safety, Galway, Ireland, June 14–18, 2004, The Marine Institute.

James, 2003, Ubiquitous ’benign’ alga emerges as the cause of shellfish contamination responsible for the human toxic syndrome, azaspiracid poisoning, Toxicon, 41, 145, 10.1016/S0041-0101(02)00244-1

Hess, 2007, Isolation and purification of azaspiracids from naturally contaminated materials, and evaluation of their toxicological effects– final project report ASTOX (ST/02/02), Marine Institute – Marine Environment & Health Series, 28, 129

Steidinger, K, Landsberg, J, Tomas, CR, and Vargo, GA (2004). Harmful Algae 2002, Florida Fish and Wildlife Conservation Commission, Florida Institute of Oceanography, and Intergovernmental Oceanographic Commission of UNESCO.

Villalba, A, Reguera, B, Romalde, JL, and Beiras, R (2003). Molluscan Shellfish Safety, Proc 4th Intl Conf Molluscan Shellfish Safety, Consellería de Pesca e Asuntos Marítimos da Xunta de Galicia and Intergovernmental Oceanographic Commission of UNESCO.

Henshilwood, K, Deegan, B, McMahon, T, Cusack, C, Keaveney, S, Silke, J, O’Cinneide, M, Lyons, D, and Hess, P Confirmation of azaspiracids in mussels in Norwegian coastal areas, and full profile at one location. The Marine Institute, Rinville, Oranmore, Galway, Ireland.

James, 2002, First evidence of an extensive northern European distribution of azaspiracid poisoning (AZP) toxins in shellfish, Toxicon, 40, 909, 10.1016/S0041-0101(02)00082-X

Lehane, 2003, The first identification of azaspiracids in shellfish from France and Spain, Toxicon, 42, 105, 10.1016/S0041-0101(03)00105-3

Maes, 2002, An outbreak of diarrhoeic shellfish poisoning in Antwerp, Belgium, Eurosurveillance, 7, 139

Vale, 2004, Is there a risk of human poisoning by azaspiracids from shellfish harvested at the Portuguese coast?, Toxicon, 44, 943, 10.1016/j.toxicon.2004.07.026

Torgersen, 2008, Structural confirmation and occurrence of azaspiracids in Scandinavian brown crabs (Cancer pagurus), Toxicon, 51, 93, 10.1016/j.toxicon.2007.08.008

Yasumoto, 2001, The chemistry and biological function of natural marine toxins, The Chemical Record, 1, 228, 10.1002/tcr.1010

Steidinger, K, Landsberg, J, Tomas, CR, and Vargo, GA (2004). Harmful Algae 2002, Florida Fish and Wildlife Conservation Commission, Florida Institute of Oceanography, and Intergovernmental Oceanographic Commission of UNESCO.

Rundberget, T, Sandvik, M, Hovgaard, P, Nguyen, L, Aasen, JAB, Castberg, T, Gustad, E, and Miles, CO (2006). Use of SPATT disks in Norway: detection of AZAs & DTXs and comparison with algal cell counts and toxin profiles in shellfish. Marine Biotoxin Science Workshop No. 23, 35–38.

Miles, 2004, A novel pectenotoxin, PTX-12, in Dinophysis spp. and shellfish from Norway, Chem Res Toxicol, 17, 1423, 10.1021/tx049870a

Cusack, C, Chamberlain, T, Devilly, L, Clarke, D, and Silke, J (2001).

Gribble, 2007, Biodiversity, biogeography and potential trophic impact of Protoperidinium spp. (Dinophyceae) off the southwestern coast of Ireland, J Plankton Res, 29, 931, 10.1093/plankt/fbm070

Poulton, 2005, Toxin variability in natural populations of Alexandrium fundyense in Casco Bay, Maine – evidence of nitrogen limitation, Deep Sea Res II, 52, 2501, 10.1016/j.dsr2.2005.06.029

Gribble, 2007, High intraindividual, intraspecific, and interspecific variability in large-subunit ribosomal DNA in the heterotrophic dinoflagellates Protoperidinium, Diplopsalis, and Preperidinium (Dinophyceae), Phycologia, 46, 315, 10.2216/06-68.1

Park, 2006, First successful culture of the marine dinoflagellate Dinophysis acuminata, Aquatic Microb Ecol, 45, 101, 10.3354/ame045101

Jeong, 2005, Feeding by red-tide dinoflagellates on the cyanobacterium Synechococcus, Aquatic Microb Ecol, 41, 131, 10.3354/ame041131

Jeong, 1999, The ecological roles of heterotrophic dinoflagellates in marine planktonic community, J Euk Microbiol, 46, 390, 10.1111/j.1550-7408.1999.tb04618.x

Ehrenreich, 2005, Distribution and diversity of natural product genes in marine and freshwater cyanobacterial cultures and genomes, Appl Environ Microbiol, 71, 7401, 10.1128/AEM.71.11.7401-7413.2005

Walsh, 2004, Polyketide and nonribosomal peptide antibiotics: modularity and versatility, Science, 303, 1805, 10.1126/science.1094318

Rundberget, 2007, Extraction of microalgal toxins by large-scale pumping of seawater in Spain and Norway, and isolation of okadaic acid and dinophysistoxin-2, Toxicon, 50, 960, 10.1016/j.toxicon.2007.07.003

MacKenzie, 2004, Solid phase adsorption toxin tracking (SPATT): a new monitoring tool that simulates the biotoxin contamination of filter feeding bivalves, Toxicon, 44, 901, 10.1016/j.toxicon.2004.08.020

Hallegraeff, GM, Blackburn, SI, Bolch, CJ, and Lewis, RJ (2001). Harmful Algal Blooms 2000, Intergovernmental Oceanographic Commission of UNESCO.

NRC-Canada, Certificate of analysis – NRC-CRM-AZA1 (lot 20060719), certified calibration solution for Azaspiracid–1 (1.57 +/− 0.09 μg/g). 2007.

Graneli, 2006, Phycotoxin pathways in aquatic food webs: Transfer, accumulation and degradation, Ecology of Harmful Algae, 189, 283, 10.1007/978-3-540-32210-8_22

Bricelj, 1998, Paralytic shellfish toxins in bivalve molluscs: occurrence, transfer kinetics, and biotransformation, Rev Fish Sci, 6, 315, 10.1080/10641269891314294

Furey, 2003, Geographical, temporal, and species variation of the polyether toxins, azaspiracids, in shellfish, Environ Sci Technol, 37, 3078, 10.1021/es020246z

Lehane, 2003, Food safety implications of distribution of azaspiracids in the tissue compartments of scallops (Pecten maximus), Food Addit Contam, 20, 154, 10.1080/0265203021000050275

James, 2002, Azaspiracid shellfish poisoning: unusual toxin dynamics in shellfish and the increased risk of acute human intoxications, Food Addit Contam, 19, 555, 10.1080/02652030210126398

Nzoughet, KJ, Hamilton, JTG, Floyd, SD, Douglas, A, Nelson, J, Devine, L, and Elliott, CT Azaspiracid: First evidence of protein binding in shellfish. Toxicon, in press.

Hess, P, McMahon, T, Slattery, D, Swords, D, Dowling, G, McCarron, M, Clarke, D, Devilly, L, Gibbons, W, Silke, J, and O’Cinneide, M Biotoxin chemical monitoring in Ireland 2001. Marine Institute, Galway, Ireland.

Yasumoto, T, Oshima, Y, and Fukuyo, Y (1996). Harmful and Toxic Algal Blooms, Intergovernmental Oceanographic Commission of UNESCO.

Hess, 2005, Tissue distribution, effect of cooking and parameters affecting the extraction of azaspiracids from mussels, Mytilus edulis, prior to analysis by liquid chromatography coupled to mass spectrometry, Toxicon, 46, 62, 10.1016/j.toxicon.2005.03.010

Hess, P, Brown, NA, Bates, LA, Turriff, JJ, Howard, FG, and Moffat, CF (2000). Fisheries Research Services, Report No 03/00.

Colman, 2005, Teratogenic effects of azaspiracid-1 identified by microinjection of Japanese medaka (Oryzias latipes) embryos, Toxicon, 45, 881, 10.1016/j.toxicon.2005.02.014

McMahon, 1996, Winter toxicity of unknown aetiology in mussels, Harmful Algae News, 14, 2

Flanagan, AF (2002). Detection and biochemical studies on the novel algal toxin, azaspiracid, National University of Ireland.

McMahon, 1998, Re-occurrence of winter toxicity, Harmful Algae News, 17, 12

Reguera, B, Blanco, J, Fernandez, ML, and Wyatt, T (1998). Harmful Algae, Santiago de Compostela: Xunta de Galicia and Intergovernmental Oceanographic Commission of UNESCO.

Pizarro, 2003, Mouse models for the study of Crohn’s disease, Trends Mol Med, 9, 218, 10.1016/S1471-4914(03)00052-2

Ito, 2000, Multiple organ damage caused by a new toxin azaspiracid, isolated from mussels produced in Ireland, Toxicon, 38, 917, 10.1016/S0041-0101(99)00203-2

Ito, 2006, Structure toxicity relationships of synthetic azaspiracid-1 and analogs in mice, Harmful Algae, 5, 586, 10.1016/j.hal.2005.11.005

Ito, 2002, Chronic effects in mice caused by oral administration of sublethal doses of azaspiracid, a new marine toxin isolated from mussels, Toxicon, 40, 193, 10.1016/S0041-0101(01)00226-4

Nicolaou, 2006, Cell growth inhibition and actin cytoskeleton disorganization induced by azaspiracid-1 structure-activity studies, Chem Res Toxicol, 19, 1459, 10.1021/tx060131z

Donato, 2003, Intracellular and extracellular roles of S100 proteins, Microsc Res Tech, 60, 540, 10.1002/jemt.10296

Twiner, 2008, Transcriptional profiling and inhibition of cholesterol biosynthesis in human lymphocyte T cells by the marine toxin azaspiracid, Genomics, 91, 289, 10.1016/j.ygeno.2007.10.015

Anderson, WA, Whelan, P, Ryan, M, McMahon, T, and James, KJ (2001). Risk assessment of azaspiracids (AZAs) in shellfish. Food Safety Authority of Ireland.

Flanagan, 1999, Azaspiracid, detection of a newly discovered phycotoxin in vitro, J Shellfish Res, 18, 716

Cohen, 1989, The structure and regulation of protein phosphatases, Annu Rev Biochem, 58, 453, 10.1146/annurev.bi.58.070189.002321

Flanagan, 2001, A cytotoxicity assay for the detection and differentiation of two families of shellfish toxins, Toxicon, 39, 1021, 10.1016/S0041-0101(00)00241-5

Twiner, 2005, Cytotoxic and cytoskeletal effects of azaspiracid-1 on mammalian cell lines, Toxicon, 45, 891, 10.1016/j.toxicon.2005.02.015

Roman, 2002, Azaspiracid-1, a potent, nonapoptotic new phycotoxin with several cell targets, Cell Signal, 14, 703, 10.1016/S0898-6568(02)00015-3

Ronzitti, 2007, Azaspiracid-1 alters the E-cadherin pool in epithelial cells, Toxicol Sci, 95, 427, 10.1093/toxsci/kfl167

Henshilwood, K, Deegan, B, McMahon, T, Cusack, C, Keaveney, S, Silke, J, O’Cinneide, M, Lyons, D, and Hess, P Development of a functional in vitro bioassay for azaspiracids (AZA) using human colonic epithelial cells. The Marine Institute, Rinville, Oranmore, Galway, Ireland.

Vale, 2007, Effects of azaspiracid-1, a potent cytotoxic agent, on primary neuronal cultures. A structure-activity relationship study, J Med Chem, 50, 356, 10.1021/jm061063g

Kulagina, 2006, Azaspiracid-1 inhibits bioelectrical activity of spinal cord neuronal networks, Toxicon, 47, 766, 10.1016/j.toxicon.2006.02.011

Artursson, 2001, Caco-2 monolayers in experimental and theoretical predictions of drug transport, Adv Drug Deliv Rev, 46, 27, 10.1016/S0169-409X(00)00128-9

Narai, 1997, Rapid decrease in transepithelial electrical resistance of human intestinal caco-2 cell monolayers by cytotoxic membrane perturbents, Toxicol Vitro, 11, 347, 10.1016/S0887-2333(97)00026-X

Nicolaou, 2007, Irreversible cytoskeletal disarrangement is independent of caspase activation during in vitro azaspiracid toxicity in human neuroblastoma cells, Biochem Pharm, 74, 327, 10.1016/j.bcp.2007.04.004

Rossini, 2001, The toxic responses induced by okadaic acid involve processing of multiple caspase isoforms, Toxicon, 39, 763, 10.1016/S0041-0101(00)00202-6

McConkey, 1998, Biochemical determinants of apoptosis and necrosis, Toxicol Lett, 99, 157, 10.1016/S0378-4274(98)00155-6

Hengartner, 1992, The biochemistry of apoptosis, Nature, 407, 770, 10.1038/35037710

Alfonso, 2001, Maitotoxin-induced calcium entry in human lymphocytes: Modulation by yessotoxin, Ca2+ channel blockers and kinases, Cell Signal, 13, 711, 10.1016/S0898-6568(01)00200-5

Alfonso, 2001, Modulation of cytosolic calcium levels of human lymphocytes by yessotoxin, a novel marine phycotoxin, Biochem Pharmacol, 61, 827, 10.1016/S0006-2952(01)00549-4

Perovic, 2000, Dinoflagellates from marine algal blooms produce neurotoxic compounds: effects on free calcium levels in neuronal cells and synaptosomes, Environ Toxicol Pharmacol, 8, 83, 10.1016/S1382-6689(99)00035-6

Xi, 1996, Glutamate receptors and calcium entry mechanisms for domoic acid in hippocampal neurons, Neuroreport, 7, 1115, 10.1097/00001756-199604260-00002

Twiner, 2005, Extracellular organic compounds from the ichthyotoxic red tide alga Heterosigma akashiwo elevate cytosolic calcium and induce apoptosis in Sf9 cells, Harmful Algae, 4, 789, 10.1016/j.hal.2004.12.006

Roman, 2004, Effects of azaspiracids 2 and 3 on intracellular cAMP, [Ca2+], and pH, Chem Res Toxicol, 17, 1338, 10.1021/tx0341862

Alfonso, 2006, Azaspiracids modulate intracellular pH levels in human lymphocytes, Biochem Biophys Res Comm, 346, 1091, 10.1016/j.bbrc.2006.06.019

Tapia, 2008, Crosstalk of tight junction components with signaling pathways, Biochim Biophys Acta, 1778, 729, 10.1016/j.bbamem.2007.08.018

Hartsock, 2008, Adherens and tight junctions: Structure, function and connections to the actin cytoskeleton, Biochim Biophys Acta, 1778, 660, 10.1016/j.bbamem.2007.07.012

Cowan, 2003, Mitogen-activated protein kinases: new signaling pathways functioning in cellular responses to environmental stress, J Exper Biol, 206, 1107, 10.1242/jeb.00220

Lambert, 2005, Depletion of Caco-2 cell cholesterol disrupts barrier function by altering the detergent solubility and distribution of specific tight-junction proteins, Biochem J, 387, 553, 10.1042/BJ20041377

Hess, P, McCarron, P, Ryan, MP, and Ryan, G (2003, January November). Isolation and purification of azaspiracids from naturally contaminated materials and evaluation of their toxicological effects (ASTOX). Proceedings of the 4th Irish Marine Biotoxin Science Workshop, Ireland.

Mileson, B, Faustman, E, Olin, S, Ryan, PB, Ferenc, S, and Burke, T (1999). International Life Science Institute Workshop Report, ILSI Press.

FSA-UK, Draft action plan on the risk assessment of mixtures of pesticides and similar substances, July, 2003. 2003.

http://www.epa.gov/pesticides/cumulative/.

Aasen, JAB (2005). PhD thesis, Oslo University.

Tubaro, 2004, Short-term oral toxicity of homoyessotoxins, yessotoxin and okadaic acid in mice, Toxicon, 43, 439, 10.1016/j.toxicon.2004.02.015

Henshilwood, K, Deegan, B, McMahon, T, Cusack, C, Keaveney, S, Silke, J, O’Cinneide, M, Lyons, D, and Hess, P (2006).

Aune, 2002, Comparison of oral and intraperitoneal toxicity of yessotoxin towards mice, Toxicon, 40, 77, 10.1016/S0041-0101(01)00192-1

Ogino, 1997, Toxicologic evaluation of yessotoxin, Nat Toxins, 5, 255, 10.1002/(SICI)1522-7189(1997)5:6<255::AID-NT6>3.0.CO;2-P

Report of the Joint FAO/IOC/WHO ad hoc Expert Consultation on Biotoxins in Molluscan Bivalves (Oslo, Norway, 26–30 September 2004). Food and Agriculture Organization.

(2004). Regulation (EC) No 853/2004 of 29 April 2004 laying down specific hygiene rules for the hygiene of foodstuffs. Off J Europ Commun, L139–55ff.

(2006, January 26–28th). Bejing China.

(2006, January 10–13th). Ottawa, Canada.

Anon, 2002, Commission Decision 225/2002/EEC, laying down detailed rules for the implementation of Council Directive 91/492/EEC as regards the maximum levels and the methods of analysis of certain marine biotoxins in bivalve molluscs, echinoderms, tunicates and marine gastropods (16.3.2002), Off J Europ Commun, L75, 62

Ofuji, 1999, A sensitive and specific determination method for azaspiracids by liquid chromatography mass spectrometry, Nat Toxins, 7, 247, 10.1002/1522-7189(199911/12)7:6<247::AID-NT68>3.0.CO;2-T

Anon., Council Directive 91/492/EEC of 15 July 1991, laying down the health conditions for the production and the placing on the market of live bivalve molluscs. 1991.

(May, January 21). Brussels, Belgium.

Anon, 1993, Council Decision 93/383/EEC of 14 June 1993 on reference laboratories for the monitoring of marine biotoxins, Off J Europ Commun, L166, 31

Hess, 2006, Three Rs Approaches in Marine Biotoxin Testing: The Report and Recommendations of a joint ECVAM/DG SANCO Workshop (ECVAM Workshop 55), Altern Labor Anim (ATLA), 32, 193, 10.1177/026119290603400207

(2005). Regulation (EC) No 2074/2005 of 5 December 2005 laying down implementing measures for certain products under Regulation (EC) No 853/2004 of the European Parliament and of the Council and for the organisation of official controls under Regulation (EC) No 854/2004 of the European Parliament and of the Council and Regulation (EC) No 882/2004 of the European Parliament and of the Council, derogating from Regulation (EC) No 852/2004 of the European Parliament and of the Council and amending Regulations (EC) No 853/2004 and (EC) No 854/2004. Off J Europ Commun, L338–27ff.

(2004). Regulation (EC) No 854/2004 of 29 April 2004 laying down specific hygiene rules for the organisation of official controls on products of animal origin intended for human consumption. Off J Europ Commun, L155–206ff.

McCarron, 2007, Effect of addition of antibiotics and an antioxidant, on the stability of tissue reference materials for domoic acid, the amnesic shellfish poison, Anal Bioanal Chem, 387, 2495, 10.1007/s00216-006-0833-3

McCarron, 2007, Freeze–drying for the stabilisation of shellfish toxins in mussel tissue (Mytilus edulis) reference materials, Anal Bioanal Chem, 387, 2475, 10.1007/s00216-006-1104-z

(25th, January 24th). Cesenatico, Italy.

Forsyth, 2006, Antibodies with broad specificity to azaspiracids by use of synthetic haptens, J Am Chem Soc, 128, 15114, 10.1021/ja066971h

Yoshida, 1997, Acute oral toxicity of microcystin-LR, a cyanobacterial hepatotoxin, in mice, Nat Toxins, 5, 91, 10.1002/nt.1

Hall, S, and Stricharts, E (1990). Marine Toxins: Origin, Structure, and Molecular Pharmacology, American Chemical Society.