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

Autor: Cécile Bernard, Rachelle Lanceleur, Marc Troussellier, Sophie Krys, Christophe Furger, Valérie Fessard, Elie Augier, Bénédicte Morin, Sahima Hamlaoui, Ronel Biré, Serge Moukha, Anne-Laure Sérandour, Sylvain Derick, Aurélie Ledreux
Přispěvatelé: Unité de Toxicologie des Contaminants, Laboratoire de Fougères - ANSES, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Molécules de Communication et Adaptation des Micro-Organismes (MCAM), Muséum national d'Histoire naturelle (MNHN)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut des Sciences Moléculaires (ISM), Centre National de la Recherche Scientifique (CNRS)-École Nationale Supérieure de Chimie et de Physique de Bordeaux (ENSCPB)-Université Sciences et Technologies - Bordeaux 1-Université Montesquieu - Bordeaux 4-Institut de Chimie du CNRS (INC), Unité de recherche Mycologie et Sécurité des Aliments (MycSA), Institut National de la Recherche Agronomique (INRA), Biotoxin Chemistry, Marine Institute, Laboratoire d'Études Rurales (LER), Université Lumière - Lyon 2 (UL2)-Isara, Ecosystèmes et Interactions Toxiques, Muséum national d'Histoire naturelle (MNHN)-Institut National de la Santé et de la Recherche Médicale (INSERM), General Directorate for food (Ministry of Agriculture), General Directorate for health (Ministry of Labor, Employment and Health), Directorate for Sea Fisheries & Aquaculture (Ministry of Agriculture), Muséum national d'Histoire naturelle (MNHN)-Centre National de la Recherche Scientifique (CNRS), Université Montesquieu - Bordeaux 4-Université Sciences et Technologies - Bordeaux 1-École Nationale Supérieure de Chimie et de Physique de Bordeaux (ENSCPB)-Centre National de la Recherche Scientifique (CNRS), MycSA, Université Lumière - Lyon 2 (UL2)
Jazyk: angličtina
Rok vydání: 2012
Předmět:
Zdroj: Analytical and Bioanalytical Chemistry
Analytical and Bioanalytical Chemistry, Springer Verlag, 2012, 403 (inpress), epub ahead of print. ⟨10.1007/s00216-012-6028-1⟩
Analytical and Bioanalytical Chemistry, Springer Verlag, 2012, epub ahead of print. ⟨10.1007/s00216-012-6028-1⟩
ISSN: 1618-2642
1618-2650
Popis: 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 IC(50) 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.
Databáze: OpenAIRE
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