{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T06:19:32Z","timestamp":1787206772180,"version":"build-2736575974"},"reference-count":63,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2020,2,21]],"date-time":"2020-02-21T00:00:00Z","timestamp":1582243200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>In the last decade, several outbreaks of ciguatera fish poisoning (CFP) have been reported in the Canary Islands (central northeast Atlantic Ocean), confirming ciguatera as an emerging alimentary risk in this region. Five Gambierdiscus species, G. australes, G. excentricus, G. silvae, G. carolinianus and G. caribaeus, have been detected in macrophytes from this area and are known to produce the ciguatoxins (CTXs) that cause CFP. A characterization of the toxicity of these species is the first step in identifying locations in the Canary Islands at risk of CFP. Therefore, in this study the toxicity of 63 strains of these five Gambierdiscus species were analysed using the erythrocyte lysis assay to evaluate their maitotoxin (MTX) content. In addition, 20 of the strains were also analysed in a neuroblastoma Neuro-2a (N2a) cytotoxicity assay to determine their CTX-like toxicity. The results allowed the different species to be grouped according to their ratios of CTX-like and MTX-like toxicity. MTX-like toxicity was especially high in G. excentricus and G. australes but much lower in the other species and lowest in G. silvae. CTX-like toxicity was highest in G. excentricus, which produced the toxin in amounts ranging between 128.2 \u00b1 25.68 and 510.6 \u00b1 134.2 fg CTX1B equivalents (eq) cell\u22121 (mean \u00b1 SD). In the other species, CTX concentrations were as follows: G. carolinianus (100.84 \u00b1 18.05 fg CTX1B eq cell\u22121), G. australes (31.1 \u00b1 0.56 to 107.16 \u00b1 21.88 fg CTX1B eq cell\u22121), G. silvae (12.19 \u00b1 0.62 to 76.79 \u00b1 4.97 fg CTX1B eq cell\u22121) and G. caribaeus (&lt;LOD to 90.37 \u00b1 15.89 fg CTX1B eq cell\u22121). Unlike the similar CTX-like toxicity of G. australes and G. silvae strains from different locations, G. excentricus and G. caribaeus differed considerably according to the origin of the strain. These differences emphasise the importance of species identification to assess the regional risk of CFP.<\/jats:p>","DOI":"10.3390\/toxins12020134","type":"journal-article","created":{"date-parts":[[2020,2,26]],"date-time":"2020-02-26T04:18:29Z","timestamp":1582690709000},"page":"134","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":40,"title":["Toxicity Characterisation of Gambierdiscus Species from the Canary Islands"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6052-9067","authenticated-orcid":false,"given":"Araceli E.","family":"Rossignoli","sequence":"first","affiliation":[{"name":"Instituto Espa\u00f1ol de Oceanograf\u00eda, Centro Ocenogr\u00e1fico de Vigo, Subida a Radiofaro 50, 36390 Vigo, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7049-9116","authenticated-orcid":false,"given":"Angels","family":"Tud\u00f3","sequence":"additional","affiliation":[{"name":"IRTA, Ctra. Poble Nou, km. 5.5, 43540 Sant Carles de la R\u00e0pita, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isabel","family":"Bravo","sequence":"additional","affiliation":[{"name":"Instituto Espa\u00f1ol de Oceanograf\u00eda, Centro Ocenogr\u00e1fico de Vigo, Subida a Radiofaro 50, 36390 Vigo, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9403-8151","authenticated-orcid":false,"given":"Patricio A.","family":"D\u00edaz","sequence":"additional","affiliation":[{"name":"Centro i~mar &amp; CeBiB, Universidad de Los Lagos, Casilla 557, Puerto Montt, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jorge","family":"Diog\u00e8ne","sequence":"additional","affiliation":[{"name":"IRTA, Ctra. Poble Nou, km. 5.5, 43540 Sant Carles de la R\u00e0pita, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pilar","family":"Riob\u00f3","sequence":"additional","affiliation":[{"name":"Department of Photobiology and Toxinology of Phytoplankton, Instituto de Investigaciones Marinas, CSIC, Eduardo Cabello 6, 36208 Vigo, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,2,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"67","DOI":"10.2331\/suisan.45.67","article-title":"Thecal structure of a marine toxic dinoflagellate Gambierdiscus toxicus gen et sp-nov collected in a ciguatera-endemic area","volume":"45","author":"Adachi","year":"1979","journal-title":"Bull. Jpn. Soc. Sci. Fish"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"739","DOI":"10.1016\/j.toxicon.2009.06.013","article-title":"Growth and toxin production in the ciguatera-causing dinoflagellate Gambierdiscus polynesiensis (Dinophyceae) in culture","volume":"56","author":"Chinain","year":"2010","journal-title":"Toxicon"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1016\/j.hal.2011.02.004","article-title":"Monitoring of dissolved ciguatoxin and maitotoxin using solid-phase adsorption toxin tracking devices: Application to Gambierdiscus pacificus in culture","volume":"10","author":"Caillaud","year":"2011","journal-title":"Harmful Algae"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/978-3-540-87895-7_1","article-title":"Marine toxins: An overview","volume":"46","author":"Fusetani","year":"2009","journal-title":"Prog. Mol. Subcell. Biol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1021","DOI":"10.2331\/suisan.43.1021","article-title":"Finding of a dinoflagellate as a likely culprit of ciguatera","volume":"43","author":"Yasumoto","year":"1977","journal-title":"Bull. Jpn. Soc. Sci. Fish"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1115","DOI":"10.1016\/0041-0101(91)90209-A","article-title":"Purification and characterization of ciguatoxins from moray eel (Lycodontis javanicus, Muraenidae)","volume":"29","author":"Lewis","year":"1991","journal-title":"Toxicon"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1543","DOI":"10.1016\/j.bcp.2005.03.022","article-title":"Azaspiracid-4 inhibits Ca2+ entry by stored operated channels in human T Iymphocytes","volume":"69","author":"Alfonso","year":"2005","journal-title":"Biochem. Pharmacol."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Ikehara, T., Kuniyoshi, K., Oshiro, N., and Yasumoto, T. (2017). Biooxidation of ciguatoxins leads to species-specific toxin profiles. Toxins, 9.","DOI":"10.3390\/toxins9070205"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"43","DOI":"10.2183\/pjab.81.43","article-title":"Chemistry, etiology, and food chain dynamics of marine toxins","volume":"81","author":"Yasumoto","year":"2005","journal-title":"Proc. Jpn. Acad. Ser B Phys. Biol. Sci."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"EFSA Panel on Contaminants in the Food Chain (2010). Scientific Opinion on marine biotoxins in shellfish\u2014Emerging toxins: Ciguatoxin group. EFSA J., 8.","DOI":"10.2903\/j.efsa.2010.1627"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"799","DOI":"10.1016\/j.toxicon.2006.07.019","article-title":"Ciguatera: Australian perspectives on a global problem","volume":"48","author":"Lewis","year":"2006","journal-title":"Toxicon"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Skinner, M.P., Brewer, T.D., Johnstone, R., Fleming, L.E., and Lewis, R.J. (2011). Ciguatera Fish Poisoning in the Pacific Islands (1998 to 2008). PLoS Negl. Trop. Dis., 5.","DOI":"10.1371\/journal.pntd.0001416"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1016\/j.hal.2014.07.007","article-title":"High abundance of the potentially maitotoxic dinoflagellate Gambierdiscus carpenteri in temperate waters of New South Wales, Australia","volume":"39","author":"Kohli","year":"2014","journal-title":"Harmful Algae"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Larsson, M.E., Laczka, O.F., Harwood, D.T., Lewis, R.J., Himaya, S.W.A., Murray, S.A., and Doblin, M.A. (2018). Toxicology of Gambierdiscus spp. (Dinophyceae) from tropical and temperate Australian Waters. Mar. Drugs, 16.","DOI":"10.3390\/md16010007"},{"key":"ref_15","unstructured":"Gouveia, N.N., Vale, P., Gouveia, N., and Delgado, J. (2009, January 12\u201315). Primeiro registo da ocorrencia de epis\u00f3dios do tipo ciguat\u00e9rico no arquip\u00e9 lago da Madeira. Proceedings of the Algas T\u00f3xicas e Biotoxinas nas aguas da Pen\u00ednsula Ib\u00e9rica 2009, IPIMAR, Lisboa, Portugal."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"6032","DOI":"10.1021\/ac100516q","article-title":"First toxin profile of Ciguateric fish in Madeira Arquipelago (Europe)","volume":"82","author":"Otero","year":"2010","journal-title":"Anal. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Costa, P.R., Estevez, P., Castro, D., Soli\u00f1o, L., Gouveia, N., Santos, C., Rodrigues, S.M., Leao, J.M., and Gago-Mart\u00ednez, A. (2018). New Insights into the occurrence and toxin Profile of Ciguatoxins in Selvagens Islands (Madeira, Portugal). Toxins, 10.","DOI":"10.3390\/toxins10120524"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"576","DOI":"10.1016\/j.scitotenv.2019.03.445","article-title":"Predictive score and probability of CTX-like toxicity in fish samples from the official control of ciguatera in the Canary Islands","volume":"673","author":"Saavedra","year":"2019","journal-title":"Sci. Total Environ."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1981","DOI":"10.3201\/eid1112.050393","article-title":"Ciguatera fish poisoning, Canary Islands","volume":"11","author":"Luzardo","year":"2005","journal-title":"Emerg. Infect. Dis."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1516","DOI":"10.1016\/j.toxicon.2010.07.021","article-title":"Ciguatera fish poisoning on the West Africa Coast: An emerging risk in the Canary Islands (Spain)","volume":"56","author":"Boada","year":"2010","journal-title":"Toxicon"},{"key":"ref_21","first-page":"2","article-title":"Appearance of ciguatoxin fish in atlantic europe waters","volume":"2","author":"Bravo","year":"2015","journal-title":"World Acad. Sci. Eng. Technol. Med Health Sci."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1016\/j.hal.2017.06.009","article-title":"Canary Islands (NE Atlantic) as a biodiversity \u2018hotspot\u2019 of Gambierdiscus: Implications for future trends of ciguatera in the area","volume":"67","author":"Fraga","year":"2017","journal-title":"Harmful Algae"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Bravo, I., Rodr\u00edguez, F., Ramilo, I., Rial, P., and Fraga, S. (2019). Ciguatera-causing dinoflagellate Gambierdiscus spp. (Dinophyceae) in a subtropical region of North Atlantic Ocean (Canary Islands): Morphological characterization and biogeography. Toxins, 11.","DOI":"10.3390\/toxins11070423"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1267","DOI":"10.1126\/science.155.3767.1267","article-title":"Ciguatoxin: Isolation and chemical nature","volume":"155","author":"Scheuer","year":"1967","journal-title":"Science"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"8929","DOI":"10.1021\/ja00206a032","article-title":"Structures of ciguatoxin and its congener","volume":"111","author":"Murata","year":"1989","journal-title":"J. Am. Chem. Soc."},{"key":"ref_26","first-page":"379","article-title":"Differential actions of pacific ciguatoxin-1 on sodium channel subtypes in mammalian sensory neurons","volume":"288","author":"Strachan","year":"1999","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"3795","DOI":"10.1038\/emboj.2012.207","article-title":"Ciguatoxins activate specific cold pain pathways to elicit burning pain from cooling","volume":"31","author":"Vetter","year":"2012","journal-title":"EMBO J."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1111\/j.1476-5381.1983.tb10485.x","article-title":"Contraction and increase in tissue calcium content induced by maitotoxin, the most potent known marine toxin, in intestinal smooth muscle","volume":"79","author":"Ohizumi","year":"1993","journal-title":"Br. J. Pharmacol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"7098","DOI":"10.1021\/ja00095a013","article-title":"Structure and partial stereochemical assignments for maitotoxin, the most toxic and largest natural Non-biopolymer","volume":"116","author":"Murata","year":"1994","journal-title":"J. Am. Chem. Soc."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/j.ejphar.2004.01.010","article-title":"Pharmacological differentiation of the P2X7 receptor and the maitotoxin-activated cationic channel","volume":"487","author":"Lundy","year":"2004","journal-title":"Eur. J. Pharmacol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1400","DOI":"10.1016\/j.toxicon.2008.03.015","article-title":"Attenuation of maitotoxin-induced cytotoxicity in rat aortic smooth muscle cells by inhibitors of Na+\/Ca2+ exchange, and calpain activation","volume":"51","author":"Frew","year":"2008","journal-title":"Toxicon"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.toxicon.2009.03.018","article-title":"Role of the sodium hydrogen exchanger in maitotoxin-induced cell death in cultured rat cortical neurons","volume":"54","author":"Wang","year":"2009","journal-title":"Toxicon"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1093\/oxfordjournals.jbchem.a122438","article-title":"Some chemical properties of maitotoxin, a putative calcium channel agonist from a marine dinoflagellate","volume":"104","author":"Yokoyama","year":"1988","journal-title":"J. Biochem."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Botana, M.L. (2014). Toxicology of seafood toxins: A critical review. Seafood and Freshwater Toxins, Pharmacology, Physiology and Detection, CRC Press.","DOI":"10.1201\/b16662"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Yasumoto, T., Hashimoto, Y., Bagnis, R., Randall, J.E., and Banner, A.H. (1971). Toxicity of the surgeonfishes. Bull. Jpn. Soc. Sci. Fish, 37.","DOI":"10.2331\/suisan.37.724"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"359","DOI":"10.2331\/suisan.42.359","article-title":"Toxicity of the surgeon fishes: II. Properties of the principal water soluble toxin","volume":"42","author":"Yasumoto","year":"1976","journal-title":"Bull. Jpn. Soc. Sci. Fish"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Kretzschmar, A.L., Verma, A., Kohli, G., and Murray, S. (2019). Development of a quantitative PCR assay for the detection and enumeration of a potentially ciguatoxin-producing dinoflagellate, Gambierdiscus lapillus (Gonyaulacales, Dinophyceae). PLoS ONE, 14.","DOI":"10.1101\/544247"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"80","DOI":"10.1016\/j.hal.2018.09.001","article-title":"Development of an LC\u2013MS\/MS method to simultaneously monitor maitotoxins and selected ciguatoxins in algal cultures and P-CTX-1B in fish","volume":"80","author":"Murray","year":"2018","journal-title":"Harmful Algae"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"886","DOI":"10.1021\/ac200799j","article-title":"Detailed LC-MS\/MS analysis of ciguatoxins revealing distinct regional and species characteristics in fish and causative alga from the Pacific","volume":"83","author":"Yogi","year":"2011","journal-title":"Anal. Chem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"731","DOI":"10.1016\/j.toxicon.2009.07.039","article-title":"Characteristic profiles of Ciguatera toxins in different strains of Gambierdiscus spp.","volume":"56","author":"Roeder","year":"2010","journal-title":"Toxicon"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"3627","DOI":"10.1002\/rcm.5264","article-title":"Preliminary study for rapid determination of phycotoxins in microalgae whole cells using matrix assisted laser desorption\/ionization time of flight mass spectrometry","volume":"25","author":"Paz","year":"2011","journal-title":"Rapid Commun. Mass Spectrom."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"804","DOI":"10.1016\/j.toxicon.2009.07.028","article-title":"Establishing a public health analytical service based on chemical methods for detecting and quantifying Pacific ciguatoxin in fish samples","volume":"56","author":"Stewart","year":"2010","journal-title":"Toxicon"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"328","DOI":"10.1016\/j.toxrep.2017.06.006","article-title":"Qualitative and quantitative assessment of the presence of Ciguatoxin, P-CTX-1B, in Spanish Mackerel (Scomberomorus commerson) from waters in New SouthWales (Australia)","volume":"4","author":"Kohli","year":"2017","journal-title":"Toxicol. Rep."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Litaker, R.W., Holland, W.C., Hardison, D.R., Pisapia, F., Hess, P., Kibler, S.R., and Tester, P.A. (2017). Ciguatoxicity of Gambierdiscus and Fukuyoa species from the Caribbean and Gulf of Mexico. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0185776"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.hal.2011.06.013","article-title":"Gambierdiscus excentricus sp. nov. (Dinophyceae): A benthic toxic dinoflagellate from the Canary Islands (NE Atlantic Ocean)","volume":"11","author":"Fraga","year":"2011","journal-title":"Harmful Algae"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1016\/j.hal.2017.02.005","article-title":"Toxicity screening of 13 Gambierdiscus strains using neuro-2a and erythrocyte lysis bioassays","volume":"63","author":"Pisapia","year":"2017","journal-title":"Harmful Algae"},{"key":"ref_47","first-page":"2447","article-title":"Assessment of cytotoxicity in ten strains of Gambierdiscus australes from Macaronesian Islands by neuro-2a cell-based assays","volume":"30","author":"Andre","year":"2018","journal-title":"J. Appl. Physiol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/j.toxicon.2012.12.016","article-title":"Differences in the toxicity of six Gambierdiscus (Dinophyceae) species measured using an in vitro human erythrocyte lysis assay","volume":"65","author":"Holland","year":"2013","journal-title":"Toxicon"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1000","DOI":"10.1080\/19440049.2012.660707","article-title":"Towards the standardisation of the neuroblastoma (neuro-2a) cell-based assay for ciguatoxin-like toxicity detection in fish: Application to fish caught in the Canary Islands","volume":"29","author":"Caillaud","year":"2012","journal-title":"Food Addit. Contam. Part A"},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Pisapia, F., Sibat, M., Herrenknecht, C., Lhaute, K., Gaiani, G., Ferron, P.J., Fessard, V., Fraga, S., Nascimento, S.M., and Litaker, R.W. (2017). Maitotoxin-4, a Novel MTX analog produced by Gambierdiscus excentricus. Mar. Drugs, 15.","DOI":"10.3390\/md15070220"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"567","DOI":"10.1080\/19440049.2012.755644","article-title":"Evaluation of seafood toxicity in the Australes archipelago (French Polynesia) using the neuroblastoma cell-based assay","volume":"30","author":"Pawlowiez","year":"2013","journal-title":"Food Addit. Contam. Part A"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"8240","DOI":"10.1038\/s41598-017-08682-8","article-title":"Identification of ciguatoxins in a shark involved in a fatal food poisoning in the Indian Ocean","volume":"7","author":"Palacios","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"5719","DOI":"10.3390\/md12125719","article-title":"Alternative methods for the detection of emerging marine toxins: Biosensors, biochemical assays and cell-based assays","volume":"12","author":"Solino","year":"2014","journal-title":"Mar. Drugs"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1016\/j.hal.2014.07.018","article-title":"Production of ciguatoxin and maitotoxin by strains of Gambierdiscus australes, G. pacificus and G. polynesiensis (Dinophyceae) isolated from Rarotonga, Cook Islands","volume":"39","author":"Rhodes","year":"2014","journal-title":"Harmful Algae"},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Munday, R., Murray, S., Rhodes, L.L., Larsson, M.E., and Harwood, D.T. (2017). Ciguatoxins and maitotoxins in extracts of sixteen Gambierdiscus isolates and one Fukuyoa isolate from the South Pacific and their toxicity to mice by intraperitoneal and oral administration. Mar. Drugs, 15.","DOI":"10.3390\/md15070208"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/j.hal.2014.05.018","article-title":"A feeding study to probe the uptake of Maitotoxin by snapper (Pagrus auratus)","volume":"37","author":"Kohli","year":"2014","journal-title":"Harmful Algae"},{"key":"ref_57","doi-asserted-by":"crossref","unstructured":"Nishimura, T., Sato, S., Tawong, W., Sakanari, H., Uehara, K., Mahfusur, M., Shah, R., Suda, S., Yasumoto, T., and Taira, Y. (2013). Genetic diversity and distribution of the Ciguatera-causing dinoflagellate Gambierdiscus spp. (Dinophyceae) in coastal areas of Japan. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0060882"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"1838","DOI":"10.3390\/md8061838","article-title":"Update on methodologies available for ciguatoxin determination: Perspectives to confront the onset of Ciguatera Fish Poisoning in Europe","volume":"8","author":"Caillaud","year":"2010","journal-title":"Mar. Drugs"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"541","DOI":"10.1016\/j.toxicon.2008.06.028","article-title":"Comparative study of the use of neuroblastoma cells (Neuro-2a) and neuroblastoma \u00d7 glioma hybrid cells (NG108-15) for the toxic effect quantification of marine toxins","volume":"52","year":"2008","journal-title":"Toxicon"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/0022-1759(83)90303-4","article-title":"Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays","volume":"65","author":"Mosmann","year":"1993","journal-title":"J. Immunol. Methods"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"190","DOI":"10.1006\/abio.1993.1476","article-title":"Tetrazolium-based cell bioassay for neurotoxins active on voltage-sensitive sodium channels: Semiautomated assay for saxitoxins, brevetoxins, and ciguatoxins","volume":"214","author":"Manger","year":"1993","journal-title":"Anal. Biochem."},{"key":"ref_62","unstructured":"R Development Core Team (2013). R: A Language and Environment for Statistical Computing, R Foundation for Statistical Computing."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1016\/j.hal.2007.09.001","article-title":"Proposal for a simple and sensitive haemolytic assay for palytoxin. Toxicological dynamics, kinetics, ouabain inhibition and thermal stability","volume":"7","author":"Paz","year":"2008","journal-title":"Harmful Algae"}],"container-title":["Toxins"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6651\/12\/2\/134\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T08:59:40Z","timestamp":1760173180000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6651\/12\/2\/134"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,2,21]]},"references-count":63,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2020,2]]}},"alternative-id":["toxins12020134"],"URL":"https:\/\/doi.org\/10.3390\/toxins12020134","relation":{},"ISSN":["2072-6651"],"issn-type":[{"value":"2072-6651","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,2,21]]}}}