GeneralDistribution OCDE/GD(97)19 REPORTOFTHEFINALRINGTESTOFTHE DAPHNIAMAGNAREPRODUCTIONTEST ORGANISATIONFORECONOMICCO-OPERATIONANDDEVELOPMENT Paris 48855 DocumentcompletdisponiblesurOLISdanssonformatd'origine CompletedocumentavailableonOLISinitsoriginalformat 2 OECDEnvironmentalHealthandSafetyPublications SeriesonTestingandAssessment No.6 ReportoftheFinalRingTestofthe DaphniamagnaReproductionTest EnvironmentDirectorate OrganisationforEconomicCo-operationandDevelopment Paris1997 3 AlsopublishedintheSeriesonTestingandAssessment: NOoE.C1D,GGuuiiddealnicneesDofocrumTeesnttinfgorotfheChDeemvieclaolspme(1n9t93o;f reformatted1995) No.2,DetailedReviewPaperonBiodegradability Testing(1995) NAos.ses3,smeGnutid(a1n9c95e)DocumentforAquatic Effects No. 4, Report of the OECD Workshop on EnvironmentalHazard/RiskAssessment(1995) No.5,ReportoftheSETAC/OECDWorkshopon AvianToxicityTesting(1996) No.7,GuidanceDocumentonDirectPhototrans- formationofChemicalsinWater(inpreparation) ComparisonofEcologicalHazard/RiskAssessment Schemes(inpreparation) GuidanceDocumentonDoseLevelSelectionin CarcinogenicityStudies(inpreparation) DetailedReviewPaperonAquaticToxicityTesting Methods(inpreparation) ©OECD1997 Applicationsforpermissiontoreproduceortranslateallorpartofthismaterialshouldbe madeto:HeadofPublicationsService,OECD,2rueAndre-Pascal,75775ParisCedex16, France. 4 AbouttheOECD The Organisation for Economic Co-operation and Deveiopment(OECD) isan intergovernmental organisationinwhichrepresentativesof29industriaiisedcountriesinNorthAmerica,Europeandthe Pacific,asweiiastheEuropeanCommission,meettoco-ordinateandharmonizepolicies,discussissues ocfarrmiuetduaoiutcboyncmeornr,eathnadnw2o0r0kstpoegoeitahieirzetdoCroemsmpiotntdeetosianntdersnuabtisoindaiiarpyrgorboiuempss.coMomsptosoefdfhoefMOeEmCbD'esrcwoournktriys delegates.ObserversfromseveralcountrieswithspecialstatusattheOECD,andfrominterested sinutbesrindaitairoynaglroorugpasniasraetisonesr,veadttbeyndthmeanOyECofDthSeecOreEtCarDi'ast,WloorckastehdopinsPaanrdis,otFhrearnmcee,etwinhgisc.hiCsoomrmgiatntieseesdainntdo DirectoratesandDivisions. TPRDhreoeoccgouwrmmoamermenkmntedso.afattniAhdosentOpseacErht(CntiohDcfealirftesorlrewapmtooeerrdrktst.oloengcTachlhheleyemmibibceicasnaltdliksntangefosetwotinynng,ioMsftechtmaehrberOseieEerdCpucDoboulutinhctaiarsniteitisohs)nes,su,EeatndshvesiwereOvoleEnlrmCaelaDnstCaTonleuunsmcHtieelaGrluotDiuhedsceialsniGidnuoeinsSds,aafnaaecnrtdeey acollectionofmethodsusedtoassessthehazardsofchemicalsandofchemicalpreparationssuchas pheesatlitchidaensdawnilddlipfhea,ramnacdeuatciccuamlusl.atTihoenyancodvedregtreasdtastifoornpihnystihcealenavinrdoncmheenmti.caTlheproOpEerCtiDesT,eseftfeGcutisdeolninheusmaarne recognisedworldwideasthestandardreferencetoolforchemicaltesting. oMnorteheiOnfEoCrDma’tsioWnoarbloduWtitdheeWEnevbirsointeme(nsteaelpHaegaelt6h).andSafetyProgrammeanditspublicationsisavailable The Environmental Health and Safety Programme co-operates closely with other international organisations.ThisdocumentwasproducedwithintheframeworkoftheInter-OrganizationProgrammefor theSoundManagementofChemicals(lOMC). 5 Thispublicationisavailabieelectronicaiiy,atnocharge. ForthecompietetextofthisandmanyotherEnvironmentai HealthandSafetypublications,consuittheOECD’s WoridWideWebsite(http://www.oecd.org/ehs/) orcontact: OECDEnvironmentDirectorate, EnvironmentaiHeaithandSafetyDivision 2rueAndre-Pascai 75775ParisCedex16 France Fax:(33-1)45241675 E-maii:[email protected] The Inter-Organization ProgrammefortheSound ManagementofChemicals(lOMC)was eOsrtgaanbilziasthieodns)in,f1o9l9l5owibnygrUeNcEoPm,meInLdOa,tiFoAnOs,mWaHdOe,byUNthIeDO199a2ndUNthCeonOfeErCeDnce(tohenPEanrvtiircoipnamteinntg andDevelopmenttostrengthenco-operationandincreaseinternationalco-ordinationinthefield ofchemicalsafety. ThepurposeofthelOMCistopromoteco-ordinationofthepoliciesand activitiespursuedbytheParticipatingOrganizations,jointlyorseparately,toachievethesound managementofchemicalsinrelationtohumanhealthandtheenvironment. 6 Foreword Thisdocumentisthereportofthe RingTestofthe February1994OECD draft Guideline202,partIIonDaphniamagnaReproductionTest,whichwasconductedin1994. DaphniaItmaalgsnoainRcelpurdoedsuctthieonreTpeosrtt,owfhitchhewOaEsCDhelWdoratksShhoefpfioelndtUhneiveFirnsailtyRoinng27T-es2t8oMfartchhe 1995(AppendixE). TheJointMeetingoftheChemicalsGroupandtheManagementCommitteeofthe Special Programme on the Control ofChemicals recommendedthatthis document be derestricted.ItisbeingpublishedontheresponsibilityoftheSecretary-GeneraloftheOECD. Authors: IanSims WRC FlenleyRoad Medmenham,Marlow BucksSL72HD UnitedKingdom PetWerRVCanDijk HenleyRoad Medmenham,Marlow BucksSL72HD UnitedKingdom JaneGamble UniversityofSheffield Sheffield S102TN UnitedKingdom NicolaGrandy OECD 2,rueAndrePascal 75775ParisCedex16 France Marie-ChantalHuet OECD 2,rueAndrePascal 75775ParisCedex16 France 7 8 TableofContents Summary 13 L Introduction 15 1.1 Ringtestobjectives 16 Background 16 1.2.1 1985EuropeanUnionringtest 16 1.2.2 Europeancollaborativeresearchprogramme 16 1.2.3 PilotRingTest 18 2. Ringtestmethods 21 2.1 Testsubstances...... 21 2.2 Instructionstoparticipants 21 2.3 Spreadsheet 21 3. Reportedtestconditions 22 3J Clone 22 3.2 Medium 23 3.2.1 Pre-testculturemedium 23 3.2.2 Ringtestmedium 23 3J3 Dietandration 24 3.4 Strategiesforneonateremoval 25 3.5 Presenceofmales 26 3.6 Testvolumes 26 3.7 Waterquality 27 3J.1 pH 27 3.7.2 Dissolvedoxygen 28 3.7.4 Totalhardness 30 3.8 Validitycriteria 30 3.9 Analysisandmaintenanceoftestsubstanceconcentrations 31 9 4. Analysisofreproductiondatafromeachparent 39 4J Statisticalmethods 39 4.1.1 DefinitionnftheNOEC 39 4.1.2 CalculationoftheNOECaccommodation 39 4.1.3 Estimationofthe21-dayEC„,EC,„andEC,„ 39 4.2 Preliminaryanalysisusingtotaljuvenilesfromeachparent 42 4.2.1 3.4-dichloroaniline 42 4.2.2 Cadmium 42 4.2.3 Phenol 43 4.3 Analysistocompensateforadultmortality 44 4.3.1 Theproblemofadultmortality 44 4.3.2 Alternativeresponsevariables 45 4.3.3 Illustrativeexample 46 4.3.4 Ringtestresults 49 4.4 Effectsoftestmedium 59 4.5 Effectsofclone 61 4.6 Effectsofration 61 4.7 Variabilitywithinandbetweenlaboratory 61 5. ComparisonofNOECandEC 78 5.1 Background 78 5.2 ComparisonofNOECandEC„intheringtest 79 5.3 UsingEC,„andslope 80 6. Conclusionsandrecommendations 84 L Literature 87 8. Acknowledgements 89 AppendixA RingTestParticipants 90 AppendixB DRreapfrtoOduEcCtiDonTeTsetstGuuisdeedlifnoer2t0he2,riPnagrtteIsItDaphniamagna 96 10