Rapid Extraction and Identification of Maitotoxin and Ciguatoxin-Like Toxins from Caribbean and Pacific Gambierdiscus Using a New Functional Bioassay

PLoS One. 2016 Jul 28;11(7):e0160006. doi: 10.1371/journal.pone.0160006. eCollection 2016.

Abstract

Background: Ciguatera is a circumtropical disease produced by polyether sodium channel toxins (ciguatoxins) that enter the marine food chain and accumulate in otherwise edible fish. Ciguatoxins, as well as potent water-soluble polyethers known as maitotoxins, are produced by certain dinoflagellate species in the genus Gambierdiscus and Fukuyoa spp. in the Pacific but little is known of the potential of related Caribbean species to produce these toxins.

Methods: We established a simplified procedure for extracting polyether toxins from Gambierdiscus and Fukuyoa spp. based on the ciguatoxin rapid extraction method (CREM). Fractionated extracts from identified Pacific and Caribbean isolates were analysed using a functional bioassay that recorded intracellular calcium changes (Ca2+) in response to sample addition in SH-SY5Y cells. Maitotoxin directly elevated Ca2+i, while low levels of ciguatoxin-like toxins were detected using veratridine to enhance responses.

Results: We identified significant maitotoxin production in 11 of 12 isolates analysed, with 6 of 12 producing at least two forms of maitotoxin. In contrast, only 2 Caribbean isolates produced detectable levels of ciguatoxin-like activity despite a detection limit of >30 pM. Significant strain-dependent differences in the levels and types of ciguatoxins and maitotoxins produced by the same Gambierdiscus spp. were also identified.

Conclusions: The ability to rapidly identify polyether toxins produced by Gambierdiscus spp. in culture has the potential to distinguish ciguatoxin-producing species prior to large-scale culture and in naturally occurring blooms of Gambierdiscus and Fukuyoa spp. Our results have implications for the evaluation of ciguatera risk associated with Gambierdiscus and related species.

MeSH terms

  • Animals
  • Biological Assay*
  • Cell Line, Tumor
  • Chromatography, High Pressure Liquid
  • Ciguatoxins / isolation & purification*
  • Dinoflagellida / chemistry*
  • Humans
  • Marine Toxins / isolation & purification*
  • Oxocins / isolation & purification*
  • Pacific Ocean
  • Spectrometry, Fluorescence

Substances

  • Marine Toxins
  • Oxocins
  • Ciguatoxins
  • maitotoxin

Grants and funding

All support for this work was provided by a NOAA Ecology and Oceanography of Harmful Algae award to PT and WL, ECOHAB contribution number 753. RJL is an NHMRC Principal Research Fellow and IV is an ARC DECRA Fellow. NHMRC grant APP1045592 to IV. There was no additional external funding received for this study.