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#1 option: Use    
single chemicals & drugs
mixtures of chemicals
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h-hours
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mortality
growth /malformation effects
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ADME
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omics methods
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other
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refinement
reduction
replacement
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#2 option: Type    
in silico
physico-chemical methods
in vitro
in vivo embryo
in vivo lower organisms
in vivo modified vertebrates
integrated test strategy
data analysis, extrapolation
other
#2 option: Status    
experimental
prevalidation
ongoing validation
validated
regulatory test
other
not relevant
#2 option: Use    
single chemicals & drugs
mixtures of chemicals
environmental samples
other
not relevant
#2 option: Exposure    
h-hours
d-days
w-weeks
hpf-hours post fertilisation
dph-days post hatch
multiple
other
not relevant
#2 option: Endpoint    
mortality
growth /malformation effects
reproduction effects
endocrine disruption
bioaccumulation, bioconcentration
ADME
PBPK
omics methods
multiple
other
not relevant
#2 option: 3R    
refinement
reduction
replacement
not relevant
#3 option: Type    
in silico
physico-chemical methods
in vitro
in vivo embryo
in vivo lower organisms
in vivo modified vertebrates
integrated test strategy
data analysis, extrapolation
other
#3 option: Status    
experimental
prevalidation
ongoing validation
validated
regulatory test
other
not relevant
#3 option: Use    
single chemicals & drugs
mixtures of chemicals
environmental samples
other
not relevant
#3 option: Exposure    
h-hours
d-days
w-weeks
hpf-hours post fertilisation
dph-days post hatch
multiple
other
not relevant
#3 option: Endpoint    
mortality
growth /malformation effects
reproduction effects
endocrine disruption
bioaccumulation, bioconcentration
ADME
PBPK
omics methods
multiple
other
not relevant
#3 option: 3R    
refinement
reduction
replacement
not relevant

 
Name Description Reference Type Status Use Exposure Endpoint 3R Standard assay link Contact link
Amphibian Luc assayAssay for detecting of the thyroid disrupting activity of tested substanceMurata and Yamauchi, Toxicol Appl Pharmacol. 2008 Feb 1;226(3):309-17in vitroexperimentalsingle chemicals & drugs
mixtures of chemicals
environmental samples
d-daysendocrine disruption
omics methods
reductionAmphibian metamorphosis (Frog) [OPPTS 890.1100]
Amphibian metamorphosis assay [OECD 231]
Amphibian TR / TTR assayAssay for detecting of the thyroid disrupting activity of tested substance (competitive binding of T3 to receptor or transport protein)Murata and Yamauchi, Toxicol Appl Pharmacol. 2008 Feb 1;226(3):309-17in vitroexperimentalsingle chemicals & drugs
mixtures of chemicals
environmental samples
d-daysendocrine disruption
omics methods
reductionAmphibian metamorphosis (Frog) [OPPTS 890.1100]
Amphibian metamorphosis assay [OECD 231]
Bird embryo cardiomyocyte in vitro assayIn vitro approach for examine effects of substance on CYP1A in the bird heartJones and Kennedy, Toxicol Sci. 2009 May;109(1):66-74in vitroexperimentalsingle chemicals & drugs
mixtures of chemicals
environmental samples
d-daysomics methodsreduction
replacement
Avian dietery toxicity test [OECD 205]
Bird embryo fibroblast in vitro assayApoptosis induced in chicken embryo fibroblast cells by the tested sampleLv and Bao, Toxicol In Vitro. 2009 Oct;23(7):1360-4in vitroexperimentalsingle chemicals & drugs
mixtures of chemicals
environmental samples
d-daysmortality
growth /malformation effects
omics methods
reduction
replacement
Avian dietery toxicity test [OECD 205]
Bird embryo hepatocyte in vitro assayEffects on mRNA expression in bird (mostly chicken) hepatocytesCrump et al., Comp Biochem Physiol C Toxicol Pharmacol. 2011 Aug;154(2):129-34in vitroexperimentalsingle chemicals & drugs
mixtures of chemicals
environmental samples
dph-days post hatchmortality
omics methods
reduction
replacement
Avian dietery toxicity test [OECD 205]
Bird embryo hepatocyte ovo assayEffects on mRNA expression in bird (mostly chicken) hepatocytesCrump et al., Comp Biochem Physiol C Toxicol Pharmacol. 2011 Aug;154(2):129-34in vivo embryoexperimentalsingle chemicals & drugs
mixtures of chemicals
environmental samples
dph-days post hatchmortality
omics methods
reduction
replacement
Avian dietery toxicity test [OECD 205]
Bird embryo lethality assayEffets on pupping success in chicken embryos in ovoCrump et al., Toxicol Sci. 2010 Jun;115(2):492-500in vivo embryoexperimentalsingle chemicals & drugs
mixtures of chemicals
environmental samples
d-daysmortality
growth /malformation effects
reduction
replacement
Avian Acute Oral Toxicity Test [OECD 223]
Bird imune toxicity in vitro assayUse of bird lymphocytes to detect immunotoxicityKote et al., J Immunotoxicol. 2006 Jul 1;3(2):101-9in vitroexperimentalsingle chemicals & drugs
mixtures of chemicals
environmental samples
d-daysmortality
growth /malformation effects
reduction
replacement
Avian dietery toxicity test [OECD 205]
Bird neuronal in vitro assayEffects on mRNA expression levels of thyroid hormon responsive genes in bird neuronal cellsVongphachan et al., Toxicol Sci. 2011 Apr;120(2):392-402in vitroexperimentalsingle chemicals & drugs
mixtures of chemicals
environmental samples
d-daysmortality
omics methods
reduction
replacement
Avian dietery toxicity test [OECD 205]
Bird QSAR acutePhysico-chemical properties and structure information is compiled to model bird acute tox by oral uptake.Toropov A & Benfenati , Eur J Med Chem. 42(5), 606-13 (2007)in silicoprevalidationsingle chemicals & drugsnot relevantmortalityreplacementAvian Acute Oral Toxicity Test [OECD 223]
CALUX assays (ER, AR, GR)Assessment of chemical (or sample) potencies to activate receptor-dependent luciferase reporter gene in mammalian cell line (mechanism of endocrine disruption via estrogen ER, androgen AR, glukocorticoid GR receptors)Legler J. et al. (1999) Toxicol Sci.48(1):55-66.in vitroexperimentalsingle chemicals & drugs
mixtures of chemicals
h-hoursendocrine disruptionreplacementLegler J.
Murk T.J.
Experimental design - fish acute toxicityImprovement of the acute toxicity test experimental design - reduction in number of animals & number of concentrations used. Statistical power remains at similar level as in the standard assay.OECD (2011). Draft Guidance and Review Documents/Monographs. Draft Fish Testing Framework document. Chapters 1-8 / Rufli H., Springer T.A. (2011). Environ. Toxicol. Chem. 30, 1006-1011.otherongoing validationsingle chemicals & drugs
environmental samples
d-daysmortalityreductionFish, Acute Toxicity Test [OECD 203]
Experimental design - fish bioconcentrationImprovement of the bioconcentration test experimental design - reduction in number of animals & number of concentrations used. Statistical power remains at similar level as in the standard assay.Springer T.A. et al. (2008) Environ Toxicol Chem. 27(11), 2271-2280 / De Wolf W. et al. (2007) Integr. Environ. Assess. Manag. 3 (1), 3-17.otherongoing validationsingle chemicals & drugs
environmental samples
d-daysbioaccumulation, bioconcentrationreductionBioconcentration: Flow-through Fish Test [OECD 305]
Experimental design - fish endocrine disruptionImprovement of the reproduction test experimental design - reduction in number of animals & number of concentrations used. Statistical power remains at similar level as in the standard assay.Bosker T. et al. (2009) Environ Toxicol Chem. 28 (11), 2386-2396..otherongoing validationsingle chemicals & drugs
environmental samples
d-daysendocrine disruptionreductionFish short term reproduction assay [OECD 229]
Exposure-based waiving (EBW)Animal testing can be avoided based on available data analysis under REACH.Vermeire et al. , Regulat Toxicol. Pharmacol 2010, 58 (3), 408-420integrated test strategynot relevantsingle chemicals & drugsmultiplereductionAcute toxicity of waste water to zebrafish eggs [ISO 15088]
Toxicity to embryos and larvae of freshwater fish [ISO 12890]
Vermeire T.
Fish BMC bioconcentrationUse of the chromatographic retention (log k) in biopartitioning micellar chromatography (BMC) to evaluate the bioconcentration factor (BCF) of pesticides in fish.Bermúdez-Saldaña et al. Chromatogr A. 2005 Jan 21;1063(1-2):153-60.in silico
physico-chemical methods
single chemicals & drugsbioaccumulation, bioconcentration
ADME
PBPK
replacement
Fish cell - steroidogenesisTesting endpoints related to steroidogenesis in various fish tissue or organ cultures (multiple endpoints possible - transcription, protein level)Schnell, S. (2009) Environ. Sci. Technol. 43, 9458-9464; Kortner, T.M., Arukwe, A. (2007) General and Comparative Endocrinology 150, 419-429.in vitroexperimentalsingle chemicals & drugs
mixtures of chemicals
h-hoursendocrine disruptionreplacementArukwe A.
Porte C.
Fish cell - VTG inductionThe use of primary hepatocytes or specific cell lines (RTH-149 and ZF-L - Christianson-Heiska and Isomaaa 2008) for assessment of estrogenic potencies - induction of vitellogen VTG is measured by various methods.Christianson-Heiska and Isomaaa. 2008 Toxicology in Vitro 22, 3, 589-597 / Björkblom C et al., Chemosphere. 2008 Oct;73(7):1064-70.in vitroexperimentalsingle chemicals & drugsd-daysendocrine disruptionreduction
Fish cell free assay - steroidogenesisMeasuring metabolism of hormones in subcellular fractions of testis of marine fish speciesFernandes, D. (2007) Aquat. Toxicol. 85(4): 258-266.physico-chemical methods
in vitro
experimentalsingle chemicals & drugs
mixtures of chemicals
h-hoursendocrine disruptionreplacementPorte C.
Fish cell line – multiple cellular and biochemical endpointsSeveral markers studied on fish cell line.Zurita et al. Toxicology in Vitro Volume 19, Issue 7, October 2005, Pages 879-886in vitroexperimentalsingle chemicals & drugsd-daysomics methods
multiple
replacement
Fish cell line biotransformation uptakeTransport of D-glucose into rainbow trout hepatoblastoma cell line RTH-149 was studied using tracer methods.Mannerström M et al., Comp Biochem Physiol A Mol Integr Physiol. 2003 Nov;136(3):779-90.in vitroexperimentalsingle chemicals & drugsotherbioaccumulation, bioconcentration
ADME
reduction
Fish cell toxicity - BF-2/luc1Development and sensitivity of the fish BF-2 cell line stably transfected with luciferase reporter gene demonstrated. The results of 0.5-6h assay were correlated with 24-96h fish LC50 valuesZhanga T. et al. (2005) Toxicology in Vitro 19 (6), 797-803in vitroexperimentalsingle chemicals & drugs
environmental samples
h-hoursmortalityreplacementFish, Acute Toxicity Test [OECD 203]
Winter M.
Fish cell toxicity - PLHC-1In vitro assessment of cytotoxicity (acute toxicity) using the cell line PLHC-1 - multiple edpoints can be measuredBabich H. et al. (2007) Chemosphere (2007) 68(8):1435-41 / Knauer K. et al. (1991) Environ Saf. 21(3):327-36.in vitroexperimentalsingle chemicals & drugs
environmental samples
d-daysmortalityreplacementWheeler J.
Fish cell toxicity - RTgillIn vitro assessment of cytotoxicity (acute toxicity) using the cell line RTgill - multiple edpoints can be measuredSchirmer K. et al. (1997) Toxicol In Vitro 11, 107-119.in vitroprevalidationsingle chemicals & drugs
environmental samples
d-daysmortalityreplacementSchirmer K.
Fish cell toxicity - RTL-W1In vitro assessment of cytotoxicity (acute toxicity) using the cell line RTL-W1 - multiple edpoints can be measuredSchnell S. et al. (2009) AQUATIC TOXICOLOGY 93 (4): 244-252in vitroprevalidationsingle chemicals & drugs
environmental samples
d-daysmortalityreplacementPorte C.
Fish cell toxicity assays (general)Endpoints for cell death using fish cell lines (multiple models).Tanneberger et al., Environ. Sci. Technol. 2010, 44, 4775–4781in vitroprevalidationsingle chemicals & drugs
environmental samples
d-daysmortalityreplacementSchirmer K.
Fish cell transcriptomic estrogenInduction of vtg1, 2, esr 1 mRNA in primary hepatocytes from zebrafish (Danio rerio) is measured.Sun et al. Induction of vtg1, 2, esr 1 mRNA in primary hepatocytes from zebrafish (Danio rerio) is meChemosphere. 2010 Feb;78(7):793-9. Epub 2009 Dec 30.asuredin vitroexperimentalsingle chemicals & drugsd-daysendocrine disruptionreduction
Fish cell ZFL - androgen reporterTesting androgenic potencies using zebrafish cell line ZFL stably transfected with luciferase reporter gene under the control of AR from sticklebackOlsson, P.E. et al. (2005) Reproductive Biology and Endocrinology 3.in vitroexperimentalsingle chemicals & drugs
mixtures of chemicals
h-hoursendocrine disruptionreplacementOlsson P.-E.
Fish cryopreserved primary hepatocytesUse of cryopreserved trout hepatocytes as an alternative to freshly isolated trout hepatocytes in a bioaccumulation assessment.Mingoia RT et al., Environ Sci Technol. 2010 Apr 15;44(8):3052-8.in vitroexperimentalsingle chemicals & drugs
mixtures of chemicals
d-daysbioaccumulation, bioconcentration
omics methods
reduction
Fish cytotoxicity - primary gill epitheliaIn vitro assessment of cytotoxicity (acute toxicity) using the primary gill epithelial cells from the rainbow trout - multiple edpoints can be measuredLilius H. et al. (1995) Toxicol In Vitro. 9(3):299-305.in vitroexperimentalsingle chemicals & drugsd-daysmortalityreplacement
Fish cytotoxicity - primary hepatocytesIn vitro assessment of cytotoxicity (acute toxicity) using the primary hepatocytes from the rainbow trout - multiple edpoints can be measuredTollefsen KE et al. (2008) Ecotoxicol Environ Saf. 69(1):64-73.in vitroexperimentalsingle chemicals & drugs
environmental samples
d-daysmortalityreplacementTollefsen K.E.
Fish estrogen modelIn the male fathead minnow (FHM, Pimephales promelas), a physiologically based computational model was developed.Ekman et al. Toxicol Sci. 2009 Jun;109(2):180-92. Epub 2009 Apr 8.in silicoexperimentalsingle chemicals & drugsnot relevantendocrine disruptionreplacement
Fish MEKCMicellar electrokinetic chromatography (MEKC) used for establishment of various ecotoxicological parameters as bioaccumulation, acute and chronic LC50 in fish.Bermúdez-Saldaña et al. J Chromatogr A. 2004 Oct 15;1052(1-2):171-80.in silico
physico-chemical methods
experimentalsingle chemicals & drugsmortality
bioaccumulation, bioconcentration
ADME
PBPK
replacement
Fish PDMS-PAMPAModified parallel artificial membrane permeability assay (PAMPA) for prediction of passive uptake and elimination rate in fish.Kwon and Escher, Environ Sci Technol. 2008 Mar 1;42(5):1787-93.physico-chemical methodsexperimentalsingle chemicals & drugsbioaccumulation, bioconcentration
ADME
replacement
Fish primary cell biotransformationBiotransformation assay in primary fish hepatocytesCravedi JP et al., Xenobiotica. 1999 May;29(5):499-509.in vitroexperimentalsingle chemicals & drugsd-daysbioaccumulation, bioconcentration
ADME
reduction
Fish primary oocytes - gene expressionTesting effects of chemicals on gene expression (mostly estrogen-related genes) in freshly isolated primary oocytes from ovaries of salmonVang, S.-H. (2007) Clinical Research in Toxicology 20, 1811-1819; Kortner, T.M., Arukwe, A. (2007) Comp. Biochem. Physiol., Part C: Toxicol. Pharmacol. 146, 569-580.in vitroexperimentalsingle chemicals & drugs
mixtures of chemicals
h-hoursendocrine disruptionreplacementArukwe A.
Fish QSAR - chronic toxicity and MOAQSAR model to predict toxicity (acute toxicity and chronic endpoints) based on assessment of toxicity modes of action at pharmaceuticalsSanderson H. (2007) 1. Bull Environ Contam Toxicol. 79(3):331-5in silicoexperimentalsingle chemicals & drugsnot relevantmortality
other
replacementSanderson H.
Fish QSAR acute - classification according to logPFish acute toxicity is predicted from log P value according to Konemann 1981 or Veith et al. 1983Konemann, H. (1981). Toxicology, 19,209-21. / Veith G.D. et al. (1983) Canadian Journal of Fisheries and Aquatic Sciences. 40, 743-748.in silicoprevalidationsingle chemicals & drugsnot relevantmortalityreplacementAcute toxicity to a freshwater fish (Brachydanio rerio) [ISO 7346]
Fish QSAR acute - classification based on multiple molecular descriptorsPhysico-chemical properties and structure information (panel of molecular decriptors) is compiled to model fish acute toxicity.Zvinavashe E. et al., Chemosphere 75(11), 1531-1538in silicoprevalidationsingle chemicals & drugsnot relevantmortalityreplacementAcute toxicity to a freshwater fish (Brachydanio rerio) [ISO 7346]
Fish, Acute Toxicity Test [OECD 203]
Fish, Acute Toxicity Test [EC No 440/2008 (C1)]
Fish QSAR acute: classification according to mechanism of actionClassification rules for chemicals according to their mechanisms of action - Pimephales promelas datasetEnoch S.J. et al. (2008) Chemosphere 73(3):243-8.in silicoexperimentalsingle chemicals & drugsnot relevantmortalityreplacementCronin M.
Fish QSAR bioconcentration ICTFish bioconcentration assessment using ICT models (Caesar model).Lombardo et al., Chemistry Central Journal 2010 4(Suppl 1):S1.in silicoprevalidationsingle chemicals & drugsnot relevantbioaccumulation, bioconcentrationreplacementBioconcentration: Flow-through Fish Test [OECD 305]
Benfenati E.
Fish tissue slices - steroidogenesisMeasuring testosterone and E2 production in slices of testes tissue (Salmo trutta) after chemical exposureMarca Pereira, M.L. et al. (2011) Aquat. Toxicol. 101, 500-511.in vitroexperimentalsingle chemicals & drugs
mixtures of chemicals
h-hoursendocrine disruptionreplacementWheeler J.
Fish tissue slices - testes (androgenicity)Measuring testosterone production in slices of testes tissue (Onkorhychus, Carassius) after chemical exposureEvanson, M., van der Kraak, G. (2001) Comparative Biochemistry and Physiology - C Toxicology and Pharmacology 130, 249-258.in vitro
other
experimentalsingle chemicals & drugs
mixtures of chemicals
h-hoursendocrine disruptionreplacement
Fish toxicity prediction from alga and Daphnia (ECOSAR)Prediction of acute fish toxicity based on the correlation between toxicity data obtained with algae and Daphnia compared to fish acute toxicityReuschenbach P. et al. ECOSAR model performance with a large test set of industrial chemicals.Department of Product Safety, BASF SE, 67056 Ludwigshafen, Germany.in silicoexperimentalsingle chemicals & drugsnot relevantmortalityreplacement
Fish toxicity prediction from algaePrediction of acute fish toxicity based on the correlation between acute toxicity data of the green alga Chlorella vulgaris and fish Pimephales promelas (96h LC50) and consideration of molecular descriptorsCronin et al. 2004 Chem Res Toxicol.17(4):545-54.in silicoexperimentalsingle chemicals & drugsnot relevantmortalityreplacementCronin M.
Fish toxicity prediction from CHO cellsPrediction of acute fish toxicity based on the correlation between acute cytotoxicity data of chinese hamster ovary CHO cells and fish acute toxicity evaluated through QSAR model for Cl-alkanesZvinavashe E. et al. (2008) Chem Res Toxicol. 21(3):739-45in silicoexperimentalsingle chemicals & drugsnot relevantmortalityreplacementMurk T.J.
Fish toxicity prediction from oligochaete dataPrediction of acute fish toxicity based on the correlation between acute toxicity data of oligochaete annelid Tubifex tubifex (3 minute assay) and fish (Pimephales promelas 96h LC50).Tichý M. et al. (2007)Altern Lab Anim 35(2):229-37.in silicoexperimentalsingle chemicals & drugsnot relevantmortalityreplacement
Fish toxicity prediction from SpirostomumPrediction of acute fish toxicity based on the correlation between acute toxicity data of ciliate Spirostomum ambiguum and fish Pimephales promelas (96h LC50).Nałecz-Jawecki G., Sawicki J. (2002) Arch Environ Contam Toxicol. 42(4):389-95.in silicoexperimentalsingle chemicals & drugsnot relevantmortalityreplacement
Fish toxicity prediction from TetrahymenaPrediction of acute fish toxicity based on the correlation between acute toxicity data of ciliate Tetrahymena pyriphormis and fish Pimephales promelas (96h LC50), and classification of chemicals according to 3 structural descriptors.Kahn I. et al. (2007) Altern Lab Anim 35(1):15-24 / Dimitrov S. et al. (2004) Environ Toxicol Chem 23(2):463-70.in silicoexperimentalsingle chemicals & drugsnot relevantmortalityreplacementBenfenati E.
Cronin M.
Gene expression analysis in zebrafish embryosAssessment of gene expression of selected genes (cyp1a, hsp or EDC-related and others) in Danio rerio embryo test; Various assays were developed such as GenDarT or MolDarT.Weil M. et al. (2009) Environ Toxicol Chem. 28(9):1970-8 / Muncke et al. 2007 Environ Toxicol 22, 185in vivo embryoexperimentalsingle chemicals & drugsd-daysmultiple
other
replacementScholz S.
ChemProp toolboxComplex toolbox for QSAR, read across and category building.Schüürmann, G., Kühne, R., Kleint, F., Ebert, R.U., Rothenbacher, C., Herth, P., 1997. A software system for automatic chemical property estimation from molecular structure, QSAR in Environmental Sciences VII. SETAC Press, Pensacola, FL, USA. SETAC Pin silicoprevalidationsingle chemicals & drugsnot relevantmultiplereplacement
Limit testToxicity of a single relatively high threshold concentration is tested, e.g. 100mg/L. For fish, 7 fish are used. In case of no mortality, no further testing is required.Details described e.g. In OECD203othervalidatedsingle chemicals & drugs
environmental samples
d-daysmortalityreductionFish, Acute Toxicity Test [OECD 203]
MDCA of endocrine disrupters in fishMulticriteria decision analysis (MDCA) of EDCs in fish has been elaborated to minimize use of full life cycle assays with fishCrane M. et al. (2010) Integr Environ Assess Manag. 6(3):378-89.integrated test strategyexperimentalsingle chemicals & drugsnot relevantnot relevantnot relevant
Medaka - transgenic for estrogensTransgenic Japanese medaka with GFP under the control of estrogen-regulated gene promoter (choriogen)Kurauchi, K. et al. (2005) Environ. Sci. Technol. 39, 2762-2768.in vivo embryoexperimentalsingle chemicals & drugsd-daysendocrine disruptionreplacement
Medaka embryo - gene expressionAssessment of estrogen-dependent genes of hormone receptors (quantitative PCR) in Japanese medaka Oryzias latipesChakraborty, T. et al. (2011) Mol. Cell. Endocrinol. 333, 47-54.in vivo embryoexperimentalsingle chemicals & drugsd-daysendocrine disruptionreplacement
Medaka embryonal toxicityToxicity assay with embryos of fish Japanese medaka - Oryzias latipes.Rakotobe L. et al. (2010) J Fish Biol. 77(4):870-8.in vivo embryoexperimentalsingle chemicals & drugs
mixtures of chemicals
d-daysmultiplereplacement
MVLN estrogenicity assayAssessment of chemical (or sample) potencies to activate estrogen receptor(ER)-dependent reporter gene in mammalian cell line MVLN (general mechanism of endocrine disruption)Freyberger A., Schmuck G. (2005) Toxicol Lett 155:1–13.in vitroexperimentalsingle chemicals & drugs
mixtures of chemicals
h-hoursendocrine disruptionreplacementBláha L.
OECD QSAR ToolboxComplex software tool combining classification of chemicals with several QSAR models available for different organisms and endpointsOECD (2007b). ENV/JM/MONO(2007)2, 154 pp. / OECD (2009) ENV/JM/MONO(2009)5, 118 pp. / OECD (2009). Effects on Biotic Systems. Test No. 231in silicovalidatedsingle chemicals & drugsnot relevantmultiplereplacementAcute toxicity to a freshwater fish (Brachydanio rerio) [ISO 7346]
Fish, Acute Toxicity Test [EC No 440/2008 (C1)]
QSAR acute toxcity - selection of compoundsThe method described selection of chemicals from the EINEC databse suitable for acute toxicity QSAR predictions; 49 classes identified.Zvinavashe E. et al. (2009) Toxicol Lett. 2009 10;184(1):67-72.in silicoexperimentalsingle chemicals & drugsnot relevantmortalityreplacementMurk T.J.
Receptor binding assays (ER, AR, TR)Assessment of chemical (or sample) potencies to bind to isolated receptors such as estrogen (ER), androgen (AR) or thyroid - indication of endocrine disruption.Loomis, A.K., Thomas, P. (1999) Biol. Reprod. 61, 51-60; Rider, C.V. et al. (2009) Environ Toxicol Chem 28, 2175-2181; Tollefsen, K., (2002) General and Comparative Endocrinology 126, 14-22.in vitroexperimentalsingle chemicals & drugs
mixtures of chemicals
h-hoursendocrine disruptionreplacementLegler J.
Murk T.J.
Tollefsen K.E.
Stepdown approachTest results (EC50) of lower organisms (algae, and daphnids) are evaluated, in the absence of fish LC50 data to derive PNECwater values.Hutchinson T., H. et al. (2003) Environ.Toxicol. Chem. 22(12), 3031-3036in vivo lower organismsvalidatedsingle chemicals & drugsd-daysmortality
growth /malformation effects
reductionBioconcentration: Flow-through Fish Test [OECD 305]
Bioconcentration: Flow-through Fish Test [EC No 440/2008 (C13)]
Hutchinson T.
Tail fin assay - amphibianBiopsies of the tail fin are used to study the resorption of the tail fin ex vivo = in vitro.
TIMESTissue metabolism simulator (TIMES) uses a heuristic algorithm to generate metabolic maps.Mekenyan et al; Curr Pharm Des. 2004;10(11):1273-93.in silicoexperimentalsingle chemicals & drugsnot relevantbioaccumulation, bioconcentration
ADME
PBPK
replacement
Yeast bioluminescence assays for endocrine disruptersAssessment of chemical (or sample) potencies to activate estrogen (ER) or androgen (AR)receptor-dependent luciferase in transgenic yeast strain (general mechanism of endocrine disruption)Leskinen P. et al. (2005) Chemosphere 61:259–266.in vitroexperimentalsingle chemicals & drugs
mixtures of chemicals
h-hoursendocrine disruptionreplacementBláha L.
YES - Yeast Estrogen ScreenAssessment of chemical (or sample) potencies to activate estrogen receptor(ER)-dependent reporter gene in transgenic yeast strain (general mechanism of endorcrine disruption)Arnold SF et al. (1996) Environ Health Perspect. 104(5):544-8.in vitroexperimentalsingle chemicals & drugs
mixtures of chemicals
h-hoursendocrine disruptionreplacementLegler J.
Murk T.J.
ZFET - flow-through systemModified zebrafish embryo developmental assay using flow-through systémLammer E. et al. (2009) Toxicol In Vitro. 23(7):1436-42.in vivo embryoexperimentalsingle chemicals & drugs
mixtures of chemicals
d-daysmultiplereplacementBraunbeck T.
ZFET - microfluidic systemModified zebrafish embryo developmental assay using microfluidic flow-through systémWielhouwer E.M. (2011) Lab Chip. 21;11(10):1815-24.in vivo embryoexperimentalsingle chemicals & drugs
mixtures of chemicals
d-daysmultiplereplacement
ZFET - thyroidTesting effects of chemicals on T4 production (immunofluorescence) in D.rerioRaldua, D., Babin, P.J. (2009) Environ. Sci. Technol. 43, 6844-6850.in vivo embryoexperimentalsingle chemicals & drugs
mixtures of chemicals
d-daysendocrine disruptionreplacement
ZFET acuteEmbryo/larvae stage, used in a 48 hr-96 hr test, visual records of 4 endpoints as indication of lethality.Lammer et al., Comp Biochem Physiol C Toxicol Pharmacol. 2009 Mar;149(2):196-209. Epub 2008 Dec 3.in vivo embryoongoing validationmixtures of chemicals
environmental samples
hpf-hours post fertilisationmortality
growth /malformation effects
replacementAcute toxicity to a freshwater fish (Brachydanio rerio) [ISO 7346]
Fish, Acute Toxicity Test [OECD 203]
Fish, Acute Toxicity Test [EC No 440/2008 (C1)]
Belanger S.
Braunbeck T.
Legler J.
Scholz S.
van der Ven L.
Witters H.
ZFET BCFDetermination of bioconcentration factors (BCF) and rate constants of lipophilic substances by biomimetic extraction using a silicone rod extraction method and prevalidated through comparison with zebrafish eggs (Danio rerio)Schreiber R et al., Chemosphere. 2009 Nov;77(7):928-33. Epub 2009 Sep 13.physico-chemical methodsexperimentalsingle chemicals & drugsd-daysbioaccumulation, bioconcentrationreplacement
ZFET bevioural responseVarious behavioural assays (photomotor response, locomotion, optokinetic response, touch-evoked response) are used to detect neurotoxic/neuroactive effects or effects on the visual system.Richards FM, J Pharmacol Toxicol Methods. 2008 Jul-Aug;58(1):50-8. Epub 2008 May 2.in vivo embryoexperimentalsingle chemicals & drugsd-daysotherreplacement
ZFET biochemical markers of chronic toxicityMicronuclei and biochemical markers (cholinesterase (ChE), glutathione S-transferase (GST) and lactate dehydrogenase (LDH) are measured.Oliveira et al. Environ Sci Pollut Res Int. 2009 Sep;16(6):679-88. Epub 2009 Mar 13.in vivo embryoexperimentalsingle chemicals & drugsd-daysomics methodsreplacement
ZFET dechorinationStandard ZFET with dechorination at 24hpf stage.Henn K and Braunbeck T., Comp Biochem Physiol C Toxicol Pharmacol. 2011 Jan;153(1):91-8. Epub 2010 Sep 30.in vivo embryoexperimentalsingle chemicals & drugs
mixtures of chemicals
environmental samples
d-daysmultiplereductionBraunbeck T.
ZFET gene expression and MOA, toxicogenomicsLarge-scale toxicogenomic screening used to unravel toxicological effects and modes of action of chemicals and ecotoxicants.Sawle et al. Toxicol Sci. 2010 Nov;118(1):128-39. Epub 2010 Aug 11in vivo embryoexperimentalsingle chemicals & drugsd-daysomics methodsreplacement
ZFET heart beatAnalysis of heart beat frequency can be used to identify compounds that interfere with heart function and/general neuromuscular function.Chakraborty et al. (2011) JOURNAL OF ENVIRONMENTAL BIOLOGY 32(2) 179-183in vivo embryoexperimentalsingle chemicals & drugs
mixtures of chemicals
environmental samples
d-daysmultiplereduction
ZFET cholinesterases carboxylesterases inhibitionSuborganismic effects were examined analysing the enzyme inhibition of cholinesterases and carboxylesterasesKüster and Altenburger, Chemosphere. 2007 Jun;68(4):751-60. Epub 2007 Feb 9.in vivo embryoexperimentalsingle chemicals & drugsd-daysomics methodsreplacement
ZFET transgenic - estrogenTransgenic zebrafish with GFP under the control of estrogen-regulated gene promoter of cyp19A1BMouriec, K. et al. (2009) Developmental Dynamics 238, 2641-2651; Chen, H. et al. (2010) Aquat. Toxicol. 96, 53-61.in vivo embryoexperimentalsingle chemicals & drugsd-daysendocrine disruptionreplacementScholz S.
ZZ_Review - endocrine endpointsReview assessing endpoint sensitivity in fish endocine distuption assays; highlighting importance of MOA-based approaches.Dang Z. et al. (2011) Toxicol Lett. 202(1):36-46.othernot relevantnot relevantnot relevantendocrine disruptionrefinement
reduction
replacement
van der Ven L.
ZZ_Review - fish and mammalian cells for acute toxicityReview of the toxicity (basal cytotoxicity in vitro) between mammalian and fish cell linesCastañoa, A., Gómez-Lechónb, M.J. (2005) Toxicology in Vitro 19 (5) 695-705othernot relevantnot relevantnot relevantnot relevantnot relevant
ZZ_Review - fish cells in ecotoxicology for different endpointsReview on the use of fish cells in ecotoxicology: report and recommendation from ECVAM workshopCastaño, A. (2003) Atla-Alternatives to Laboratory Animals 31(3): 317-351.in vitronot relevantnot relevantnot relevantmultiplereplacement
ZZ_Review - reference chemicalsDevelopment of a list of reference chemicals for testing alternatives to whole fish toxicity test - 60 chemicals with different MOAs selected.Schirmer K. et al. (2008) Aquat Toxicol. 90(2):128-37.othernot relevantnot relevantotherendocrine disruptionreduction
ZZ_Review - vertebrate cells for regulatory testingProposal to improve vertebrate cell cultures to establish them as substitutes for the regulatory testing of chemicals and effluents using fish.Schirmer, K. (2006). Toxicology 224, 163-183.othernot relevantsingle chemicals & drugs
mixtures of chemicals
environmental samples
not relevantmultiplereplacementSchirmer K.
ZZ_Review fish and aquatic alternative testsReview tests based on fish cells and cell lines, fish embryos, lower organisms, and the many expert systems and QSARs for aquatic toxicity testing.Grindon et al. Altern Lab Anim. 2006 Dec;34(6):651-64.othernot relevantnot relevantnot relevantnot relevantnot relevant
ZZ_Review fish cell estrogenCritical review about current knowledge on the VTG response of cultured fish hepatocytes.Navas and Segner. Aquat Toxicol. 2006 Oct 25;80(1):1-22. Epub 2006 Sep 1.in vitronot relevantnot relevantnot relevantendocrine disruptionnot relevant
ZZ_Review ZFET, chronic toxicity, MOAReview fish embryo tests beyond the acute appliccation (omics, chronic toxicity, EDC etc.)Scholz et al. Environ Sci Pollut Res Int. 2008 Jul;15(5):394-404. Epub 2008 Jun 25. Review.in vivo embryonot relevantnot relevantd-daysendocrine disruption
bioaccumulation, bioconcentration
ADME
omics methods
multiple
refinement
replacement
Scholz S.