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Regul Toxicol Pharmacol. 2014 Aug;69(3):496-511. doi: 10.1016/j.yrtph.2014.05.018. Epub 2014 May 27.

OECD validation study to assess intra- and inter-laboratory reproducibility of the zebrafish embryo toxicity test for acute aquatic toxicity testing.

Author information

1
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.
2
University of Heidelberg, COS, Aquatic Ecology & Toxicology, Im Neuenheimer Feld 230, 69120 Heidelberg, Germany. Electronic address: rubenstrecker@gmx.de.
3
The Procter & Gamble Company, Global Product Stewardship, Mason, OH 45040, USA. Electronic address: rawlings.jm@pg.com.
4
The Procter & Gamble Company, Global Product Stewardship, Mason, OH 45040, USA. Electronic address: belanger.se@pg.com.
5
University of Heidelberg, COS, Aquatic Ecology & Toxicology, Im Neuenheimer Feld 230, 69120 Heidelberg, Germany. Electronic address: braunbeck@uni-hd.de.
6
The Procter & Gamble Company, Mason Quantitative Sciences, OH 45040, USA. Electronic address: carr.gj@pg.com.
7
VU University Amsterdam, Institute for Environmental Studies, De Boelelaan 1087, 1081 HV Amsterdam, The Netherlands. Electronic address: peter.cenijn@vu.nl.
8
Institute of Industrial Organic Chemistry Branch Pszczyna, Doswiadczalna 27, 43-200 Pszczyna, Poland. Electronic address: fochtman@ipo-pszczyna.pl.
9
Organisation for Economic Co-operation and Development, 2, rue André Pascal, 75775 Paris Cedex 16, France. Electronic address: anne.gourmelon@oecd.org.
10
Merck KGaA, Frankfurter Str. 250, 64293 Darmstadt, Germany. Electronic address: nicole.huebler@merckgroup.com.
11
European Commission Joint Research Centre, Institute for Health and Consumer Protection, via E. Fermi 2749, 21027 Ispra (VA), Italy. Electronic address: akleens1@jhu.edu.
12
UFZ - Helmholtz Centre for Environmental Research, 04318 Leipzig, Germany. Electronic address: melanie.knoebel@eawag.ch.
13
Federal Environment Agency (UBA), Versuchsfeld Marienfelde, Schichauweg 58, 12307 Berlin, Germany. Electronic address: carola.kussatz@uba.de.
14
VU University Amsterdam, Institute for Environmental Studies, De Boelelaan 1087, 1081 HV Amsterdam, The Netherlands. Electronic address: juliette.legler@vu.nl.
15
Norwegian Institute for Water Research (NIVA), Gaustadalléen 21, 0349 Oslo, Norway. Electronic address: adam.lillicrap@niva.no.
16
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.
17
Federal Environment Agency (UBA), Versuchsfeld Marienfelde, Schichauweg 58, 12307 Berlin, Germany. Electronic address: christian.polleichtner@uba.de.
18
Institute of Industrial Organic Chemistry Branch Pszczyna, Doswiadczalna 27, 43-200 Pszczyna, Poland. Electronic address: ew@ipo-pszczyna.pl.
19
BASF SE, 67056 Ludwigshafen, Germany. Electronic address: edward.salinas@basf.com.
20
BASF SE, 67056 Ludwigshafen, Germany. Electronic address: katharina.schneider@cos.uni-heidelberg.de.
21
UFZ - Helmholtz Centre for Environmental Research, 04318 Leipzig, Germany. Electronic address: stefan.scholz@ufz.de.
22
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.
23
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.
24
Federal Environment Agency (UBA), P.O. 1406, 06813 Dessau-Roßlau, Germany. Electronic address: susanne.walter-rohde@uba.de.
25
Merck KGaA, Frankfurter Str. 250, 64293 Darmstadt, Germany. Electronic address: stefan.weigt@merckgroup.com.
26
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.
27
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.

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.

KEYWORDS:

Alternative method; Aquatic acute toxicity; Reproducibility; Validation; Zebrafish embryo

PMID:
24874798
DOI:
10.1016/j.yrtph.2014.05.018
[Indexed for MEDLINE]
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