title: Development and validation of an OECD reproductive toxicity test guideline with the pond snail Lymnaea stagnalis (Mollusca, Gastropoda).
abstract: The OECD test guideline development program has been extended in 2011 to establish a partial life-cycle protocol for assessing the reproductive toxicity of chemicals to several mollusk species, including the great pond snail Lymnaea stagnalis. In this paper, we summarize the standard draft protocol for a reproduction test with this species, and present inter-comparison results obtained in a 56-day prevalidation ring-test using this protocol. Seven European laboratories performed semi-static tests with cultured snails of the strain Renilys® exposed to nominal concentrations of cadmium chloride (from 53 to 608μgCdL(-1)). Cd concentrations in test solutions were analytically determined to confirm accuracy in the metal exposure concentrations in all laboratories. Physico-chemical and biological validity criteria (namely dissolved oxygen content >60% ASV, water temperature 20±1°C, control snail survival >80% and control snail fecundity >8 egg-masses per snail over the test period) were met in all laboratories which consistently demonstrated the reproductive toxicity of Cd in snails using the proposed draft protocol. Effect concentrations for fecundity after 56days were reproducible between laboratories (68<EC50-56d<124μgL(-1)) and were consistent with literature data. EC50-56d and EC10-56d values were comprised within a factor of 1.8 and 3.6, respectively, which is in the range of acceptable variation defined for reference chemicals in OECD test guidelines for invertebrates. The inter-laboratory reproducibility coefficient of variation (CV) for the Cd LC50-56d values was 8.19%. The inter-laboratory comparison of fecundity within the controls gave a CV of 29.12%, while exposure to Cd gave a CV of 25.49% based on the EC50-56d values. The OECD has acknowledged the success of this prevalidation exercise and a validation ring-test involving 14 laboratories in Europe, North- and South-America is currently being implemented using four chemicals (Cd, prochloraz, trenbolone and tributyltin).
authors.affiliation: INRA, UMR 0985 Ecologie et Santé des Ecosystèmes, Equipe Ecotoxicologie et Qualité des Milieux Aquatiques, 65 rue de Saint-Brieuc, CS 84215, F-35042 Rennes Cedex, France. Electronic address: Virginie.ducrot@rennes.inra.fr.
authors.forename: Virginie
authors.initials: V
authors.lastname: Ducrot
authors.affiliation: CEFAS Lowestoft Laboratory, Pakefield Road, Lowestoft NR33 0HT, United Kingdom.
authors.forename: Clare
authors.initials: C
authors.lastname: Askem
authors.affiliation: INRA, Unité Expérimentale d'Ecologie et d'Ecotoxicologie, 65 rue de Saint-Brieuc, CS 84215, F-35042 Rennes Cedex, France.
authors.forename: Didier
authors.initials: D
authors.lastname: Azam
authors.affiliation: Goethe University Frankfurt am Main, Department of Aquatic Ecotoxicology, Biological Sciences Division, Max-von-Laue-Str. 13, 60348 Frankfurt, Germany.
authors.forename: Denise
authors.initials: D
authors.lastname: Brettschneider
authors.affiliation: Brixham Environmental Laboratory, AstraZeneca, Freshwater Quarry, Brixham, Devon TQ5 8BA, United Kingdom.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Brown
authors.affiliation: Université de Lyon, F-69000 Lyon, France; Université Lyon 1, France; CNRS, UMR5558, Laboratoire de Biométrie et Biologie Évolutive, F-69622 Villeurbanne, France; Institut Universitaire de France, 103 bd Saint-Michel, 75005 Paris, France.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Charles
authors.affiliation: INRA, Unité Expérimentale d'Ecologie et d'Ecotoxicologie, 65 rue de Saint-Brieuc, CS 84215, F-35042 Rennes Cedex, France.
authors.forename: Maïra
authors.initials: M
authors.lastname: Coke
authors.affiliation: INRA, UMR 0985 Ecologie et Santé des Ecosystèmes, Equipe Ecotoxicologie et Qualité des Milieux Aquatiques, 65 rue de Saint-Brieuc, CS 84215, F-35042 Rennes Cedex, France.
authors.forename: Marc
authors.initials: M
authors.lastname: Collinet
authors.affiliation: Université de Lyon, F-69000 Lyon, France; Université Lyon 1, France; CNRS, UMR5558, Laboratoire de Biométrie et Biologie Évolutive, F-69622 Villeurbanne, France; VetAgro Sup Campus Vétérinaire de Lyon, F-69280 Marcy l'Etoile, France.
authors.forename: Marie-Laure
authors.initials: ML
authors.lastname: Delignette-Muller
authors.affiliation: Université de Lyon, F-69000 Lyon, France; Université Lyon 1, France; CNRS, UMR5558, Laboratoire de Biométrie et Biologie Évolutive, F-69622 Villeurbanne, France.
authors.forename: Carole
authors.initials: C
authors.lastname: Forfait-Dubuc
authors.affiliation: Department of Biology, University of Southern Denmark, Campusvej 55, 5230 Odense M, Denmark.
authors.forename: Henrik
authors.initials: H
authors.lastname: Holbech
authors.affiliation: CEFAS Weymouth Laboratory, Barrack Road, Weymouth DT4 8UB, United Kingdom.
authors.forename: Thomas
authors.initials: T
authors.lastname: Hutchinson
authors.affiliation: BASF SE, Crop Protection - Ecotoxicology, Speyerer Strasse 2, D-67117 Limburgerhof, Germany.
authors.forename: Arne
authors.initials: A
authors.lastname: Jach
authors.affiliation: Department of Biology, University of Southern Denmark, Campusvej 55, 5230 Odense M, Denmark.
authors.forename: Karin L
authors.initials: KL
authors.lastname: Kinnberg
authors.affiliation: INRA, Unité Expérimentale d'Ecologie et d'Ecotoxicologie, 65 rue de Saint-Brieuc, CS 84215, F-35042 Rennes Cedex, France.
authors.forename: Cédric
authors.initials: C
authors.lastname: Lacoste
authors.affiliation: Brixham Environmental Laboratory, AstraZeneca, Freshwater Quarry, Brixham, Devon TQ5 8BA, United Kingdom.
authors.forename: Gareth
authors.initials: G
authors.lastname: Le Page
authors.affiliation: Old School House, Brow Edge, Backbarrow, Ulverston, Cumbria LA128QX, United Kingdom.
authors.forename: Peter
authors.initials: P
authors.lastname: Matthiessen
authors.affiliation: Goethe University Frankfurt am Main, Department of Aquatic Ecotoxicology, Biological Sciences Division, Max-von-Laue-Str. 13, 60348 Frankfurt, Germany.
authors.forename: Jörg
authors.initials: J
authors.lastname: Oehlmann
authors.affiliation: Brixham Environmental Laboratory, AstraZeneca, Freshwater Quarry, Brixham, Devon TQ5 8BA, United Kingdom.
authors.forename: Lynsey
authors.initials: L
authors.lastname: Rice
authors.affiliation: CEFAS Weymouth Laboratory, Barrack Road, Weymouth DT4 8UB, United Kingdom.
authors.forename: Edward
authors.initials: E
authors.lastname: Roberts
authors.affiliation: Goethe University Frankfurt am Main, Department of Aquatic Ecotoxicology, Biological Sciences Division, Max-von-Laue-Str. 13, 60348 Frankfurt, Germany.
authors.forename: Katharina
authors.initials: K
authors.lastname: Ruppert
authors.affiliation: CEFAS Weymouth Laboratory, Barrack Road, Weymouth DT4 8UB, United Kingdom.
authors.forename: Jessica Elphinstone
authors.initials: JE
authors.lastname: Davis
authors.affiliation: CEFAS Weymouth Laboratory, Barrack Road, Weymouth DT4 8UB, United Kingdom.
authors.forename: Clemence
authors.initials: C
authors.lastname: Veauvy
authors.affiliation: BASF SE, Crop Protection - Ecotoxicology, Speyerer Strasse 2, D-67117 Limburgerhof, Germany.
authors.forename: Lennart
authors.initials: L
authors.lastname: Weltje
authors.affiliation: CEFAS Lowestoft Laboratory, Pakefield Road, Lowestoft NR33 0HT, United Kingdom.
authors.forename: Ruth
authors.initials: R
authors.lastname: Wortham
authors.affiliation: INRA, UMR 0985 Ecologie et Santé des Ecosystèmes, Equipe Ecotoxicologie et Qualité des Milieux Aquatiques, 65 rue de Saint-Brieuc, CS 84215, F-35042 Rennes Cedex, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Lagadic
mesh_terms: D000818:Animals; D002104:Cadmium / Q000633:toxicity*; D017408:Guidelines as Topic; D008195:Lymnaea / Q000187:drug effects* / Q000502:physiology; D066150:Organisation for Economic Co-Operation and Development; D015203:Reproducibility of Results; D012098:Reproduction / Q000187:drug effects; D018675:Toxicity Tests / Q000379:methods
chemical_list: D002104:Cadmium
keywords: Fecundity; Mollusk; OECD Conceptual Framework for Testing and Assessment of Endocrine Disrupters; OECD test guidelines