title: OECD validation study to assess intra- and inter-laboratory reproducibility of the zebrafish embryo toxicity test for acute aquatic toxicity testing.
abstract: The OECD validation study of the zebrafish embryo acute toxicity test (ZFET) for acute aquatic toxicity testing evaluated the ZFET reproducibility by testing 20 chemicals at 5 different concentrations in 3 independent runs in at least 3 laboratories. Stock solutions and test concentrations were analytically confirmed for 11 chemicals. Newly fertilised zebrafish eggs (20/concentration and control) were exposed for 96h to chemicals. Four apical endpoints were recorded daily as indicators of acute lethality: coagulation of the embryo, lack of somite formation, non-detachment of the tail bud from the yolk sac and lack of heartbeat. Results (LC50 values for 48/96h exposure) show that the ZFET is a robust method with a good intra- and inter-laboratory reproducibility (CV<30%) for most chemicals and laboratories. The reproducibility was lower (CV>30%) for some very toxic or volatile chemicals, and chemicals tested close to their limit of solubility. The ZFET is now available as OECD Test Guideline 236. Considering the high predictive capacity of the ZFET demonstrated by Belanger et al. (2013) in their retrospective analysis of acute fish toxicity and fish embryo acute toxicity data, the ZFET is ready to be considered for acute fish toxicity for regulatory purposes.
authors.affiliation: European Commission Joint Research Centre, Institute for Health and Consumer Protection, via E. Fermi 2749, 21027 Ispra (VA), Italy. Electronic address: caat-eu-policy@uni-konstanz.de.
authors.forename: François
authors.initials: F
authors.lastname: Busquet
authors.affiliation: University of Heidelberg, COS, Aquatic Ecology & Toxicology, Im Neuenheimer Feld 230, 69120 Heidelberg, Germany. Electronic address: rubenstrecker@gmx.de.
authors.forename: Ruben
authors.initials: R
authors.lastname: Strecker
authors.affiliation: The Procter & Gamble Company, Global Product Stewardship, Mason, OH 45040, USA. Electronic address: rawlings.jm@pg.com.
authors.forename: Jane M
authors.initials: JM
authors.lastname: Rawlings
authors.affiliation: The Procter & Gamble Company, Global Product Stewardship, Mason, OH 45040, USA. Electronic address: belanger.se@pg.com.
authors.forename: Scott E
authors.initials: SE
authors.lastname: Belanger
authors.affiliation: University of Heidelberg, COS, Aquatic Ecology & Toxicology, Im Neuenheimer Feld 230, 69120 Heidelberg, Germany. Electronic address: braunbeck@uni-hd.de.
authors.forename: Thomas
authors.initials: T
authors.lastname: Braunbeck
authors.affiliation: The Procter & Gamble Company, Mason Quantitative Sciences, OH 45040, USA. Electronic address: carr.gj@pg.com.
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Carr
authors.affiliation: VU University Amsterdam, Institute for Environmental Studies, De Boelelaan 1087, 1081 HV Amsterdam, The Netherlands. Electronic address: peter.cenijn@vu.nl.
authors.forename: Peter
authors.initials: P
authors.lastname: Cenijn
authors.affiliation: Institute of Industrial Organic Chemistry Branch Pszczyna, Doswiadczalna 27, 43-200 Pszczyna, Poland. Electronic address: fochtman@ipo-pszczyna.pl.
authors.forename: Przemyslaw
authors.initials: P
authors.lastname: Fochtman
authors.affiliation: Organisation for Economic Co-operation and Development, 2, rue André Pascal, 75775 Paris Cedex 16, France. Electronic address: anne.gourmelon@oecd.org.
authors.forename: Anne
authors.initials: A
authors.lastname: Gourmelon
authors.affiliation: Merck KGaA, Frankfurter Str. 250, 64293 Darmstadt, Germany. Electronic address: nicole.huebler@merckgroup.com.
authors.forename: Nicole
authors.initials: N
authors.lastname: Hübler
authors.affiliation: European Commission Joint Research Centre, Institute for Health and Consumer Protection, via E. Fermi 2749, 21027 Ispra (VA), Italy. Electronic address: akleens1@jhu.edu.
authors.forename: André
authors.initials: A
authors.lastname: Kleensang
authors.affiliation: UFZ - Helmholtz Centre for Environmental Research, 04318 Leipzig, Germany. Electronic address: melanie.knoebel@eawag.ch.
authors.forename: Melanie
authors.initials: M
authors.lastname: Knöbel
authors.affiliation: Federal Environment Agency (UBA), Versuchsfeld Marienfelde, Schichauweg 58, 12307 Berlin, Germany. Electronic address: carola.kussatz@uba.de.
authors.forename: Carola
authors.initials: C
authors.lastname: Kussatz
authors.affiliation: VU University Amsterdam, Institute for Environmental Studies, De Boelelaan 1087, 1081 HV Amsterdam, The Netherlands. Electronic address: juliette.legler@vu.nl.
authors.forename: Juliette
authors.initials: J
authors.lastname: Legler
authors.affiliation: Norwegian Institute for Water Research (NIVA), Gaustadalléen 21, 0349 Oslo, Norway. Electronic address: adam.lillicrap@niva.no.
authors.forename: Adam
authors.initials: A
authors.lastname: Lillicrap
authors.affiliation: Instituto Politécnico Nacional, Escuela Nacional de Ciencias Biológicas, Lab. de Hidrobiología Experimental, Prol. Carpio esq. Plan de Ayala S/N, Col. Santo Tomas, México, D.F. 11340, Mexico. Electronic address: ferjeronimo@hotmail.com.
authors.forename: Fernando
authors.initials: F
authors.lastname: Martínez-Jerónimo
authors.affiliation: Federal Environment Agency (UBA), Versuchsfeld Marienfelde, Schichauweg 58, 12307 Berlin, Germany. Electronic address: christian.polleichtner@uba.de.
authors.forename: Christian
authors.initials: C
authors.lastname: Polleichtner
authors.affiliation: Institute of Industrial Organic Chemistry Branch Pszczyna, Doswiadczalna 27, 43-200 Pszczyna, Poland. Electronic address: ew@ipo-pszczyna.pl.
authors.forename: Helena
authors.initials: H
authors.lastname: Rzodeczko
authors.affiliation: BASF SE, 67056 Ludwigshafen, Germany. Electronic address: edward.salinas@basf.com.
authors.forename: Edward
authors.initials: E
authors.lastname: Salinas
authors.affiliation: BASF SE, 67056 Ludwigshafen, Germany. Electronic address: katharina.schneider@cos.uni-heidelberg.de.
authors.forename: Katharina E
authors.initials: KE
authors.lastname: Schneider
authors.affiliation: UFZ - Helmholtz Centre for Environmental Research, 04318 Leipzig, Germany. Electronic address: stefan.scholz@ufz.de.
authors.forename: Stefan
authors.initials: S
authors.lastname: Scholz
authors.affiliation: National Institute for Public Health and the Environment (RIVM), Centre for Environmental Quality, 3720 BA Bilthoven, The Netherlands. Electronic address: evert-jan.van.den.brandhof@rivm.nl.
authors.forename: Evert-Jan
authors.initials: EJ
authors.lastname: van den Brandhof
authors.affiliation: National Institute for Public Health and the Environment (RIVM), Centre for Health Protection, 3720 BA Bilthoven, The Netherlands. Electronic address: leo.van.der.ven@rivm.nl.
authors.forename: Leo T M
authors.initials: LT
authors.lastname: van der Ven
authors.affiliation: Federal Environment Agency (UBA), P.O. 1406, 06813 Dessau-Roßlau, Germany. Electronic address: susanne.walter-rohde@uba.de.
authors.forename: Susanne
authors.initials: S
authors.lastname: Walter-Rohde
authors.affiliation: Merck KGaA, Frankfurter Str. 250, 64293 Darmstadt, Germany. Electronic address: stefan.weigt@merckgroup.com.
authors.forename: Stefan
authors.initials: S
authors.lastname: Weigt
authors.affiliation: Flemish Institute for Technological Research (VITO), Environmental Risk & Health, Team Applied Bio & Molecular Systems (ABS), Boeretang 200, 2400 Mol, Belgium. Electronic address: hilda.witters@vito.be.
authors.forename: Hilda
authors.initials: H
authors.lastname: Witters
authors.affiliation: European Commission Joint Research Centre, Institute for Health and Consumer Protection, via E. Fermi 2749, 21027 Ispra (VA), Italy. Electronic address: marlies.halder@ec.europa.eu.
authors.forename: Marlies
authors.initials: M
authors.lastname: Halder
mesh_terms: D000818:Animals; D007753:Laboratories; D007928:Lethal Dose 50; D066150:Organisation for Economic Co-Operation and Development; D015203:Reproducibility of Results; D023382:Toxicity Tests, Acute / Q000379:methods*; D014874:Water Pollutants, Chemical / Q000633:toxicity*; D015027:Zebrafish
chemical_list: D014874:Water Pollutants, Chemical
keywords: Alternative method; Aquatic acute toxicity; Reproducibility; Validation; Zebrafish embryo