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Collaborative study for the detection of toxic compounds in shellfish extracts using cell-based assays. Part I: screening strategy and pre-validation study with lipophilic marine toxins

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Abstract

Human poisoning due to consumption of seafood contaminated with phycotoxins is a worldwide problem, and routine monitoring programs have been implemented in various countries to protect human consumers. Following successive episodes of unexplained shellfish toxicity since 2005 in the Arcachon Bay on the French Atlantic coast, a national research program was set up to investigate these atypical toxic events. Part of this program was devoted to fit-for-purpose cell-based assays (CBA) as complementary tools to collect toxicity data on atypical positive-mouse bioassay shellfish extracts. A collaborative study involving five laboratories was conducted. The responses of human hepatic (HepG2), human intestinal (Caco2), and mouse neuronal (Neuro2a) cell lines exposed to three known lipophilic phycotoxins—okadaic acid (OA), azaspiracid-1 (AZA1), and pectenotoxin-2 (PTX2)—were investigated. A screening strategy composed of standard operating procedures and a decision tree for dose–response modeling and assay validation were designed after a round of “trial-and-error” process. For each toxin, the shape of the concentration–response curves and the IC50 values were determined on the three cell lines. Whereas OA induced a similar response irrespective of the cell line (complete sigmoid), PTX2 was shown to be less toxic. AZA1 induced cytotoxicity only on HepG2 and Neuro2a, but not on Caco2. Intra- and inter-laboratory coefficients of variation of cell responses were large, with mean values ranging from 35 to 54 % and from 37 to 48 %, respectively. Investigating the responses of the selected cell lines to well-known toxins is the first step supporting the use of CBA among the panel of methods for characterizing atypical shellfish toxicity. Considering these successful results, the CBA strategy will be further applied to extracts of negative, spiked, and naturally contaminated shellfish tissues.

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Acknowledgments

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 & Aquaculture (Ministry of Agriculture) and by the General Directorate for health (Ministry of Labor, Employment and Health). The reviewers are acknowledged for their helpful comments on the manuscript.

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Authors and Affiliations

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

    Anne-Laure Sérandour, Rachelle Lanceleur & Valérie Fessard

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

    Aurélie Ledreux, Sahima Hamlaoui & Cécile Bernard

  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. Laboratoire de Toxicologie, Université Bordeaux Segalen-INRA, 146, rue Léo Saignat, 33076, Bordeaux, France

    Elie Augier & Serge Moukha

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

    Ronel Biré & Sophie Krys

  7. UMR 5119 ECOSYM Université Montpellier 2 & 1-CNRS-IRD-Ifremer, 34095, Montpellier, France

    Marc Troussellier

Authors
  1. Anne-Laure Sérandour
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  2. Aurélie Ledreux
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  3. Bénédicte Morin
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  4. Sylvain Derick
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  6. Rachelle Lanceleur
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  7. Sahima Hamlaoui
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  8. Serge Moukha
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  9. Christophe Furger
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  10. Ronel Biré
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  11. Sophie Krys
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  12. Valérie Fessard
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  13. Marc Troussellier
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  14. Cécile Bernard
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Corresponding author

Correspondence to Cécile Bernard.

Additional information

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

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Sérandour, AL., Ledreux, A., Morin, B. et al. Collaborative study for the detection of toxic compounds in shellfish extracts using cell-based assays. Part I: screening strategy and pre-validation study with lipophilic marine toxins. Anal Bioanal Chem 403, 1983–1993 (2012). https://doi.org/10.1007/s00216-012-6028-1

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  • DOI: https://doi.org/10.1007/s00216-012-6028-1

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