ISO 04979-2023.docx

INTERNATIONA1.STANDARDISO4979editionFirst2O23-O6Waterqua1.ityAquatictoxicitytestbasedonrootre-growthin1.emnaminorQUHitedeeauEssaidetoxicitaquatiquebassurIarepoussedesracineschez1.emnaminorCOPYRIGHTPROTECTEDDOCUMENT©ISO2023IUirhM*hedbdi1.iUedotherwiseupdhi.o啪InPSd1.Mc;GeatrOn0*tuuc11Dnt10tU%qn0*即b11o即PcXHiWKmt1.onmaytheinternetoranintranet,withoutpriorwrittenpermission.PermissioncanberequestedfromeitherISOattheaddressbe1.oworISO'smemberbodyinthecountr)oftherequester.f),WV>fifiU81.andonnet8CH-1214Vernier,GenevaPhone：M1.22749O1.11觥ftte:丽丽BQrgPub1.ishedinSwitzer1.andContentsForewordivIntroductionv2 Scope13 Normativereferences14 Termsanddefinitions16 Testorganisms3Growthmedium36.1 Preparationofstockso1.ution37 6.2Storageandcu1.tivation48 VPParatUS4,Experimenta1.methods58.1 1PrCPara口OnofIDedIUt1.)58.2 Preparationoftoxicantso1.utionandtestdi1.utions68.2.1 3,doiiwtUftistes1.concen1.ratians68.3 Contro1.68.4 T*rDsforOftestOrgan】SIDS.69 1."es5Oneffects79.1 Referencechcmica1.s792StatISt1.CS/Expressionofresu1.ts810.1Testresu1.tsB1110.2Expressionofresu1.ts8Testreport8Annex(informative)Rootexcisionandre-growth1.engthmeasurement10Annex(informativc)test12Bib1.iography一.一.14ForewordISO(theInternationa1.OrganizationforStandardization)isawor1.dwidefederationofnationa1.standardsttiugh(ISOmrtm匕0)bMdmi)rwcekdHPfff1.QbbGgbodyI1tf1.nmtiJMiai6sndrifc3owhtd1.ysQcchit2committeehasbeenestab1.ishedhastherighttoberepresentedonthatcommittee.Internationa1.organizations,governmenta1.andnon-governmenta1.,in1.iaisonwithISO,a1.sotakepartinthework.ISOco1.1.aboratesc1.ose1.ywiththee1.ectrotechnica1.standardization.Internationa1.E1.ectrotechnica1.Commission(IEC)ona1.1.mattersofTheproceduresusedtodeve1.opthisdocumentandthoseintendedforitsfurthermaintenanceare蜩喇搬of!SO1.E(tetheeditoria1.ru1.esoftheISO/IECDirectives.Part2(secwww.iso.org/direc1.ives).AnytradeusedinthisR«0nrights.dteE1.n殷的招舞ns副¥神吸融浒用qH1doc业我棉曲S双邯服.subjectofanypatentrightsidentifiedduringthedeve1.opmen13th»dOa向的twi1.1.beintheIntroductionand/orontheISO1.istofpatentdec1.arationsreceived(seewww.iso.org/patents).constitutenameendorsement,documentisinformationgivenfortheconvenienceofusersanddoesnotForanexp1.anationofthevo1.untarynatureofstandards,themeaningofISOspecifictermsand觐牌U那dOwarWSFMO)assE麻皿噩inwef像际R裂沁nBar池然MdeadPaes地w¼w.iso.oiso/forewoId.htm).早腋gicd带出*6脏WaSpreparedbyTechnica1.CommitteeISO/TC147.Waterqua1.itytSubcommitteeSC5,nyfeedbackorquestionsonthisdocumentshou1.dbedirectedtotheuser,snationa1.standardsbody.Acomp1.ete1.istingofthesebodiescanbefoundatwww.iso.org/members.1.Hm1.Introductionsevera1.standard20079:2005,U.S.haveEnvironmenta1.Protectionintemationa1.StandardizationEconomicexposuredurationsofat1.east7dtodetecttoxicity.rootsbyprovidingStabi1.itysensitivctoenvironmenta1.stressorspaidthatIhcyrootsimportanteco1.ogica1.However,thetoecotoxico1.ogica1.significanceeco1.ogica1.1.ysignificantendpointre-eva1.uatedandWith1.engthstandardizedmethodsre-growthbioassaydiffersinsevera1.keyaspectsfromthreeinternationa1.1.yb) thetestvesse1.isa24-we1.1.ce1.1.p1.ate;d)rootstechniqueexcisedexcisingexposuretosubsequentmeasurementsnorequirementdeve1.opedpre-se1.ectArtificia1.severanceofprevious1.y.Uoweverraccordingintonatura1.settingssincerootabsdssiondistributcdtheirdispersionsurviva1.potentia1.1.ybetter(environment.Therefore,rapidendpointabscissionre-growthconsideredorgans,we1.1.knownnonethc1.essimportantwithoutanchorage,nutrientabsorptionandnonessentia1.inappropriate.managementwastewaterIoxicitymadescrecningtime1.ymannerintheoperationa1.unexpectcdofpo1.1.utioneventsincegrowthrate/biomassendpoints.minor,butitcana1.sobeapp1.iedtoother1.emnaspeciesandSpirode1.aspecieswithsomemodifications.©ISO2023-A1.1.rightsreservedv1.emnagibhaand1.minorhavebeenmostextensive1.yusedinphytotoxicitytestingandthereare一forexamp1.e,ISOmethodswhich-beenadoptedbymajorAgency,Organizationforagencies.CooperationandDeve1.opment.Testswithduckweedhavetypica1.1.yfavouredmeasurementsoffrond(e.g.theirnumber,biomass,area,carbonuptake,ch1.orophy1.1.content(3j.(4)andrequirestandardOntheotherhand,testsbasedonroote1.ongationaresomeofthemostwide1.yusedphytotoxicitymethodo1.ogiesforterrestria1.angiospermsbecauseoftheirsimp1.icityandrapidity.Despitereportsthatro1.esof1.minorarehigh1.y5U6j.pj.(8jrhasbeenandtop1.ay1.emnasinceitwasgenera1.1.yconsideredthatrootfragi1.itymadetheirhand1.ingformeasurementsdifficu1.tandthatitwasimpractica1.toobtainsufficientnumbersofindividua1.p1.antswithidentica1.root1.engthstoinitiatetests.wasshownbesensitive,preciseandoftherootendpointhasbeenincomparisonrootmoretraditiona1.frondgrowthorbiomassendpoints.Theproposedroot(ISO,OECDandUSEP):a)thetestcanbecomp1.etedwithin72h;c) therequiredvo1.umeoftestwatersamp1.esis3m1.;ThewereofpriorrootspriorWithexposurcmeansthatthereisonnew1.ytoroots,rootsofuniform1.ength,whichreducesthehand1.ingofthesefragi1.eroots.hasnotbeenreportedrootsWiHpossib1.yneverhappenrecentstudies,thetinyg1.oba1.1.yin1.emnawaterfemsofthegenusAzo1.1.a1.osttheirrootsunderstressconditions(9japhenomenonknovrnasrapidrootabscission.Suchsheddingsetsitsfrondsfreefromroot-entang1.edmatsandfaci1.itatesanimportanttostrategyofAzo1.1.a9jThiscanindicatethattherootrootishasitseco1.ogica1.re1.evance.Itisa1.sohutarethat1.eninawi1.1.thriveforp1.antanyroots.Thus,1.emnaappearcytokininbiosynthesis.Therefore,themanipu1.ationofrootsbysimp1.eseverancecanbeanunimportantissueanddoesnotjustifytheconc1.usionthattheremova1.ofrootspriortoCcotoxico1.ogica1.testingisThethree-dayrootre-growthtestisusefu1.frtherapidscreeningofeitherwastewatereff1.uentsorhazardouscontaminantsinnatura1.WaterS网asitiseasytoperform,quickto11nandcost-effectivetooperatefordecisionshou1.dbeaandcanhaveancaseofbenefittestingtimeTherootregrowthendpointfromthis72-hourprotoco1.isnotadirectsubstitutefortheseven-dayThepresentprotoco1.providesdetai1.edinformationonhowtosetupandconducttherootre-growthtestwith1.emnaminoraswe1.1.ashowtoana1.ysetoxicitydata.Thisprotoco1.isintendedforusewith1.emnaWaterqua1.ityAquatictoxicitytestbasedonrootregrowthin1.emnaminorWARNINGPersonsusingthisdocumentshou1.dbefami1.iarwithnorma1.1.aboratorypractice.Thisdocumentdoesnotpurporttoaddressa1.1.ofthesafetyprob1.ems,ifany,associatedwithitsuse.Itistheresponsibi1.ityoftheusertoestab1.ishappropriatesafetyandhea1.thpractices.IMPORTANTItisabso1.ute1.yessentia1.thattestsconductedaccordingtothisdocumentbecarriedoutbysuitab1.ytrainedstaff.1 Scopeenvironmenta1.substancesinc1.udingtreatedmunicipa1.wastewaterandindustria1.eff1.uents.app1.iesto2 Normativereferences3 Termsanddef1.nitionsnatthefo1.1.owingISOandIECmaintaintermino1.ogica1.databasesforUSeinStandardiZatiGaddresses：3.1 IECE1.ectropedia:avai1.ab1.eath1.1.pgwww.e1.e<tropedia.orgaxeniccu1.tureSOURCE:ISO20079:2005,3.1coefficientofvariationCV3.3co1.onySOURCE:ISO20079:2005,3.4effectiveCOnCentrationEG5OUKCE:isoNUU/9：NUg3.v.modtticd-Note1toentrydeetedj©ISO2023-A)rightsreservedThisdocumentspecifiesamethodforthedeterminationoftheinhibitionofrootre-growthinduckweeds(1.emna)bywatersamp1.esandmixturescontainedorwater.ThismethodTherearenonormativereferencesinthisdocument.Forthepurposesofthisdocument,thefo1.1.owingtermsanddefinitionsapp1.y.-ISOOn1.inebrowsingp1.atform:avai1.ab1.eathttps:/www.iso.org/obpmonocu1.turesoforganismsfromasing1.especies,freefromfungi,a1.gaeandothermacrophytespecies3.2re1.ativestandarddeviation,expressedasapercentageaggregateofmotheranddaughterfronds,attachedtoeachother,sometimesreferredtoasap1.ant3.4 concentrationoftestsamp1.eatwhichaneffectofx%ismeasured,ifcomparedtothecontro1.3.5 frondindividua1.Ieaf-Iikestructureonaduckweedco1.ony;thesma1.1.estunit(i.e.individua1.),capab1.etoreproduce兜URCE:ISO20079:2005,3.10growthincreaseinbiomassovertimeastheresu1.tofpro1.iferationofnewtissuesNote1toentry:Inthistest,itreferstoanyparameterofobservation.更夕URCE:ISO20079:2005,3.13growthmedium§igewatertowhichreagent-gradechemica1.s(micronutrients)havebeenaddednon-axeniccu1.turemonocu1.tureoforganismsfromasing1.especies(i.e.freefromothermacrophytespecies),whichhasnotbeentreatedwithantimycoticorantibioticso1.utionstoremovenatura1.1.yassociatedbacteriaand驷包pre-cu1.turecu1.tureofduckweedusedforacc1.imationoftestp1.antstothetestconditionsandforthegrowingofthep1.antstobeusedintheinocu1.umOURCE:ISO20079:2005,3.19purewaterdeionizedordisti1.1.edWater更?RCE:ISO19827:2016,3.4rootp1.antpartthatnorma1.1.ygrowsunderwaterandho1.dsthep1.antinp1.ace融RCE:ISO20079:2005,3.20stockcu1.turecu1.tureofasing1.especiesofduckweedtoconservetheorigina1.1.emnaspeciesinthe1.aboratoryandtoprovideinocu1.umfortheprecu1.turcNote1.taxonomy.entry:Itisnecessarytousedefinedandverifiedstrains,becauseofpossib1.einsecuritiesinspeciesSOURCE:ISO20079:2005,3.21,modified,Anaddress1.istofsupp1.iersisgiveninAnnexC,hasbeenfromNote1toentry.stockso1.utionso1.utionwithaccurate1.yknownana1.yteconcentration(三),preparedfromchemica1.sWithanappropriatepurity(SOURCE:ISO5667-16:2017,3.213.14testsamp1.ediscreteportionofsamp1.e(takenfrom,forexamp1.e,receivingwater,wastewater,disso1.vedchemica1.substancesormixtures,productsandcompounds)pre-treatedaccordingtotheneedsofthetest(e.g.disso1.ution,fi1.tering,neutra1.ization)更ygRCE:ISO20079:2005,3.24testmediumaqueousso1.utionthatconsistsofaparticu1.arconcentrationofpreparedtestsamp1.emixtureoftestwaterandthesamp1.eundertest4SOBRiR3p1.e21427-1:2006,3.4&三t处前Ag桃KWWm熊拼WIbhdS叫谢由wi1.1.bemeasured.W蝴te网颂Wto触蝴ib曲外丽那触硼酬i蜘腑expressedastheECrh5 TestorganismsThestandardtestorganismofthistestisduckweed,1.emnaminortwhichisafreshwater-f1.oatingp1.ant.6 Growthmedium6.1 Preparationofstockso1.utionPreparestockso1.utionbyaddingtheweighedchemica1.saccordingtoTab1.e1tothedesiredvo1.umeof0i1.mioB«fH(bedurt1.tQjdiumspoundso1.utions.Purewatershou1.dbeusedforthepHof1.iquidmediasha1.1.beadjustedto6,9±0,1afteraddingpurewatertoeachstockso1.ution.Nutrientsshou1.dbeaddedintheorderOfI-II-IiI-IV-Vwhi1.epreparingtheso1.utionstopreventprecipitation.MO,11Hthctop<i6StijdthsIJSorCgrmmbPfi(Jam即titgn旭Inn1.iSQIthhetto>grhedium.appropriateIJquidmediacanbestoredforuptoonemonthatroomtemperatureinthedark.Tab1.e1MainingredientsOfSteinbergmediumStockso1.utionChemica1.sStockconcentrationFina1.concentrationMacroe1.ements81mi1.I1.MgSO7HO520IIIQ(N3)24H214,7520Tab1.e1(continued)Stockso1.utionChemica1.sStockconcentrationFina1.concentrationMicroc1.cmcntsgmi1.VFeC1.6HO0.76Na2-EDTA-ZH2O1.S6.2Storageandcu1.tivationCu1.turefrondsof1.emnaminoringrowthmedium(3.7)isshowninTab1.e!.Thefronds(3.5)sha1.1.bPmHHwtati(函01.k)tt2yW2M()ThBnairtirwoiissh出RMgntfp1.hoerinmhYUjnSit?d3创nu>1.thestoragecu1.turecanbekeptcontinua1.1.yun1.essuncontro1.1.edcontaminationoccurs.C1.oudymediumina1.emnaminorstockcu1.tureindicatesbacteria1.contamination,whereascontaminationwithmou1.dispossib1.yoras1.ime1.ayerdeve1.opsonthevesse1.CondfiXtede5Hf7"n用OrIargS8Wf1.iFaSshfiapjRf1.scarded.medium7ApparatusThetestrequiresusua1.1.aboratoryequipmentandthefo1.1.owing.7.1UniformTemperature-Contro1.1.edcabinettherequirementsUuwewent1.ight,providing,continuous,Tab1.e2Summarytestconditionsforthe1.emnarootre-growthtestTypeoftest:1.ighting:Static;72htest90mo1.msto100mo1.ms(6contro1.rep1.icatesarerecommended)Endpoint：Root1.ength曲?into出於4R¾i¾H施EHfesrd婚OWdft(A?Wftym1.1.)8rr8fiftuecyraturerangewasdetermined.As1.hnnngcoti1.JiD1.sS1.1.tH1.bCrefi)6mcndddrtrduxtptabUnESkiidmH1.aSCirafuMMedudte)MicamtfihiSOh)jigth0,199mg1.to0,475mg11.1.1.ed用他跳Eft1.di醍凫由EJBH11H¾s.1.engthoftheregrownrootsinthecontro1.sandtheECsova1.uerange9.4PrecisionTheinter1.aboratoryvariabi1.ityfrom1.emnatoxicitytestusingareferencetoxicant(CuS4-GA-107E5Ba弼iti?OHTfeSU1.tSrepeatabi1.ityand27,2%forreproducibi1.ity(see-Aftnex-BrTab1.eB.1.).10.1 Testresu1.tsRecordroot1.engthva1.uesofeachrep1.icate,meanva1.uesandstandarddeviations.Ca1.cu1.atethePetftiiiMitrfriziinqtndi1.iihbiIitpitttodtxntdeviationexpressedasapercentageofthemean,toestimatetheP1.ottingconcentrationresponsecurvesishigh1.yrecommendedtoprovidethebasisfordeterminingJh制ysvEh9tedredQk88'S(EIfifJd网fi1.espQndingstandarddeviationandCVsshaI1.beTheva1.uesshou1.dbegivenasmgIifortestswithindividua1.chemica1.soraspercentageforeff1.uent理哦力eport由自解KWimginformationandthedataandtestendpointestimatesa) thetestmethodused,togetherwithareferencetothisdocument,i.e.ISO4979:2023;b) nameofthe1.aboratoryperformingthetest;c) recentreferencesubstancetesting；d) dateandperiodoftest;e) testorganisms(e.g.scientificname,strain,source,ho1.dingconditions);0testdetai1.s; cu1.turingapparatusandincubationprocedure; cu1.turetypes(static,static-renewa1.etc.); compositionofmedium; preparationoftestsamp1.e(e.g.pH,sa1.inityofeff1.uentsamp1.e)andtreatments; concentrationstested; rep1.icatesperconcentration;AnnexA(informative)Rootexcisionandre-growth1.engthmeasurement.1.ExcisionprocedureFigureA.1.showsanexcisionprocedurefo1.1.owedbyre-growthQfrootsin1.emnamin>91088910iso1.atefrondsIiftupthefrondscuttherootroot1.ess1.eavesroot1.ess1.eavesafter12hcu1.tureregrownrootafter24hcu1.turererownrootafter48hcu1.tureregrownrootafter72hcu1.turetopviewsideviewFigureA.1.Rootexcisionandre-growth.2Root1.engthmeasurementsInFigureA.2pane1.（八）showstherootsstraightenedontheg1.asss1.ide,(b)showsanimageana1.yserfr<gi½andSCwJrtBkWU1.1.Qtiu不。$6即£ot1.tJbuj0<u1.itroJo1.!>tot)iJ<M>(Uhgwrt1.oid1.(tittthurcdFigure.2Root1.engthmeasurementsAnnexB(informative)Inter1.aboratoryprecisionofcontro1.va1.uesandECsofromthe1.emnatoxicitytestTab1.eB.1.Inter1.aboratoryprecisionofcontro1.va1.ues(min)ofroot1.engthin1.emna1.aboratory'Mean±95%C1.CV(%)1.ab1.41,4±7,616,2095%P1."Repeatabi1.ity(%)Reproduc1.b1.1.1.ty(%)1.abi:IncheonNationa1.University(Korea)1.ab2:KoreaCoastGuardMetropo1.itanPo1.ice(Korea)1.ab3:Nationa1.InstituteOfEnvtronmentaIResearch(Korea)I.ab4:InstituteofPuh1.icHea1.thandEnvironmentReMfarch(Korea)1.ab5:GhentUniversity(Be1.gium)1.ab6:ShanghaiOceanUniversity(Giina)1.ab7:UniversityofConnecticut(USA)1.ab8:Friedrich-Schi1.1.er-University,ena(Ge