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Collaborative study for the detection of toxic compounds in shellfish extracts using cell-based assays. Part II: application to shellfish extracts spiked with lipophilic marine toxins

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Abstract

Successive unexplained shellfish toxicity events have been observed in Arcachon Bay (Atlantic coast, France) since 2005. The positive mouse bioassay (MBA) revealing atypical toxicity did not match the phytoplankton observations or the liquid chromatography−tandem mass spectrometry (LC−MS/MS) investigations used to detect some known lipophilic toxins in shellfish. The use of the three cell lines (Caco2, HepG2, and Neuro2a) allows detection of azaspiracid-1 (AZA1), okadaic acid (OA), or pectenotoxin-2 (PTX2). In this study, we proposed the cell-based assays (CBA) as complementary tools for collecting toxicity data about atypical positive MBA shellfish extracts and tracking their chromatographic fractionation in order to identify toxic compound(s). The present study was intended to investigate the responses of these cell lines to shellfish extracts, which were either control or spiked with AZA1, OA, or PTX2 used as positive controls. Digestive glands of control shellfish were extracted using the procedure of the standard MBA for lipophilic toxins and then tested for their cytotoxic effects in CBA. The same screening strategy previously used with pure lipophilic toxins was conducted for determining the intra- and inter-laboratory variabilities of the responses. Cytotoxicity was induced by control shellfish extracts whatever the cell line used and regardless of the geographical origin of the extracts. Even though the control shellfish extracts demonstrated some toxic effects on the selected cell lines, the extracts spiked with the selected lipophilic toxins were significantly more toxic than the control ones. This study is a crucial step for supporting that cell-based assays can contribute to the detection of the toxic compound(s) responsible for the atypical toxicity observed in Arcachon Bay, and which could also occur at other coastal areas.

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Acknowledgements

This work was carried out as part of the Arcachon National Program funded by the General Directorate for Food and the Directorate for Sea Fisheries and Aquaculture (Ministry of Agriculture) and by the General Directorate for Health (Ministry of Labor, Employment and Health). The authors wish to thank Stéphanie Magny, of the Food Safety Laboratory of the French Agency for Food, Environmental and Occupational Health and Safety, for preparing and sending the shellfish extracts to the different labs involved in the study.

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Authors and Affiliations

  1. UMR 7245 MNHN-CNRS MCAM, Muséum National d’Histoire Naturelle, 75231, Paris, France

    Aurélie Ledreux, Sahima Hamlaoui & Cécile Bernard

  2. Unité de Toxicologie des Contaminants, ANSES, 35302, Fougères, France

    Anne-Laure Sérandour, Rachelle Lanceleur & Valérie Fessard

  3. UMR 5805 Université Bordeaux 1-CNRS EPOC, LPTC, 33405, Talence, France

    Bénédicte Morin

  4. Novaleads, 10 Avenue de l’Europe, 31520, Ramonville Saint-Agne, France

    Sylvain Derick & Christophe Furger

  5. Unité de Caractérisation des Toxines, ANSES, 94706, Maisons-Alfort, France

    Ronel Biré & Sophie Krys

  6. UMR 5119 ECOSYM Université Montpellier 2 and 1-CNRS-IRD-Ifremer, 34095, Montpellier, France

    Marc Troussellier

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  1. Aurélie Ledreux
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  2. Anne-Laure Sérandour
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  12. Cécile Bernard
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Correspondence to Cécile Bernard.

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Aurélie Ledreux and Anne-Laure Sérandour contributed equally to the publication.

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Ledreux, A., Sérandour, AL., Morin, B. et al. Collaborative study for the detection of toxic compounds in shellfish extracts using cell-based assays. Part II: application to shellfish extracts spiked with lipophilic marine toxins. Anal Bioanal Chem 403, 1995–2007 (2012). https://doi.org/10.1007/s00216-012-6029-0

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  • DOI: https://doi.org/10.1007/s00216-012-6029-0

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