1、BRITISH STANDARD BS EN 13098:2001 Workplace atmosphere Guidelines for measurement of airborne micro-organisms and endotoxin The European Standard EN 13098:2001 has the status of a British Standard ICS 13.040.30 NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBS EN 13098:2001 Th
2、is British Standard, having been prepared under the direction of the Environment Sector Committee, was published under the authority of the Standards Committee and comes into effect on 15 July 2001 BSI 07-2001 ISBN 0 580 37873 X National foreword This British Standard is the official English languag
3、e version of EN 13098:2001. The UK participation in its preparation was entrusted by Technical Committee EH/2, Air quality, to Subcommittee EH/2/2, Workplace atmospheres, which has the responsibility to: A list of organizations represented on this subcommittee can be obtained on request to its secre
4、tary. Cross-references The British Standards which implement international or European publications referred to in this document may be found in the BSI Standards Catalogue under the section entitled “International Standards Correspondence Index”, or by using the “Find” facility of the BSI Standards
5、 Electronic Catalogue. A British Standard does not purport to include all the necessary provisions of a contract. Users of British Standards are responsible for their correct application. Compliance with a British Standard does not of itself confer immunity from legal obligations. aid enquirers to u
6、nderstand the text; present to the responsible European committee any enquiries on the interpretation, or proposals for change, and keep the UK interests informed; monitor related international and European developments and promulgate them in the UK. Summary of pages This document comprises a front
7、cover, an inside front cover, the EN title page, pages 2 to 24, an inside back cover and a back cover. The BSI copyright date displayed in this document indicates when the document was last issued. Amendments issued since publication Amd. No. Date CommentsEUROPEANSTANDARD NORMEEUROPENNE EUROPISCHENO
8、RM EN13098 September2000 ICS07.100.99;13.040.30 Englishversion WorkplaceatmosphereGuidelinesformeasurementof airbornemicroorganismsandendotoxin AtmosphresdeslieuxdetravailRglespourle mesuragedemicroorganismesetdendotoxineen suspensiondanslair ArbeitsplatzatmosphreLeitlinienfrdieMessungvon Mikroorgan
9、ismenundEndotoxininderLuft ThisEuropeanStandardwasapprovedbyCENon17August2000. CENmembersareboundtocomplywiththeCEN/CENELECInternalRegulationswhichstipulatetheconditionsforgivingthisEurope an Standardthestatusofanationalstandardwithoutanyalteration.Uptodatelistsandbibliographicalreferencesconcerning
10、s uchnational standardsmaybeobtainedonapplicationtotheCentralSecretariatortoanyCENmember. ThisEuropeanStandardexistsinthreeofficialversions(English,French,German).Aversioninanyotherlanguagemadebytra nslation undertheresponsibilityofaCENmemberintoitsownlanguageandnotifiedtotheCentralSecretariathasthe
11、samestatusast heofficial versions. CENmembersarethenationalstandardsbodiesofAustria,Belgium,CzechRepublic,Denmark,Finland,France,Germany,Greece, Iceland,Ireland,Italy,Luxembourg,Netherlands,Norway,Portugal,Spain,Sweden,SwitzerlandandUnitedKingdom. EUROPEANCOMMITTEEFORSTANDARDIZATION COMITEUROPENDENO
12、RMALISATION EUROPISCHESKOMITEEFRNORMUNG CentralSecretariat:ruedeStassart,36B1050Brussels 2000CEN Allrightsofexploitationinanyformandbyanymeansreserved worldwideforCENnationalMembers. Ref.No.EN13098:2000EPage2 EN13098:2000 Contents Page Foreword 3 Introduction 4 1 Scope. 4 2 Normativereferences . 4 3
13、 Termsanddefinitions. . 4 4 Symbolsandabbreviations. 7 5 Measurementofmicroorganismsandendotoxin 7 6 Sampling . 8 7 Analyticalmethod. 10 8 Expressionofresults. 13 9 Report 13 AnnexA(informative)Recommendationsforselectionofmeasurementmethods. 14 AnnexB(informative)Exampleofsamplingform . 20 AnnexC(i
14、nformative)Listofgeneralmedia . 21 AnnexD(informative)ColonycountingFormulaandexamples. 22 Bibliography. 24Page3 EN13098:2000 Foreword ThisEuropeanStandardhasbeenpreparedbyTechnicalCommitteeCEN/TC137“Assessmentofworkplace exposure“,thesecretariatofwhichisheldbyDIN. ThisEuropeanStandardshallbegiventh
15、estatusofanationalstandard,eitherbypublicationofanidenticaltext orbyendorsement,atthelatestbyMarch2001,andconflictingnationalstandardsshallbewithdrawnatthe latestbyMarch2001. AccordingtotheCEN/CENELECInternalRegulations,thenationalstandardsorganizationsofthefollowing countriesareboundtoimplementthis
16、EuropeanStandard:Austria,Belgium,CzechRepublic,Denmark,Finland, France,Germany,Greece,Iceland,Ireland,Italy,Luxembourg,Netherlands,Norway,Portugal,Spain,Sweden, SwitzerlandandtheUnitedKingdom. ThisEuropeanStandardhasbeenpreparedbyTechnicalCommitteeCEN/TC137“Assessmentofworkplace exposure“,thesecreta
17、riatofwhichisheldbyDIN. AnnexesA,B,CandDareinformative. Thisstandardincludesabibliography. .Page4 EN13098:2000 Introduction Assessingoccupationalexposuretoairbornemicrobialcontaminantsinarepresentativewayisachallengingtask.Itis necessaryhoweverthatinformationcanbegatheredtoevaluateandtominimiseexpos
18、uretobiologicalagents.The samplingequipmentusedoftenintroducesitsowncriticallimitations,asintheassessmentofaerosolfractions.Some samplingequipmentmaybecapableonlyofmeasuringculturablemicroorganisms,whileothersallowthe characterisationofboththetotalnumberoforganismsandtheculturablefraction.Analytical
19、proceduresmayaddfurther tothedifficultiesandtheuncertainties,e.g.themethodusedmaynotallowtheidentificationofthebiologicalagents present,ormaycauseunwantedinterferencebetweendifferentbiologicalagents.However,byadheringtotheprinciples outlinedinthisstandardforchoiceofsamplingandanalyticalprocedures,th
20、eseuncertaintiescanbereducedandcon trolled,allowingcomparableandrepresentativemeasurementstobemade. 1 Scope TheEuropeanStandardprovidesguidelinesfortheassessmentofworkplaceexposuretoairbornemicroorganisms includingthedeterminationoftotalnumberandculturablenumberofmicroorganismsintheworkplaceatmosphe
21、re.The standardalsoprovidesmethodsformeasurementofairborneendotoxinintheworkenvironment. TheEuropeanStandarddoesnotapplytoviruses,specificpathogenicmicroorganismsandtoxinsotherthanendotoxin, althoughsomeofthemeasurementprinciplesmaybethesame. 2 Normativereferences ThisEuropeanStandardincorporatesbyd
22、atedorundatedreference,provisionsfromotherpublications.Thesenormative referencesarecitedattheappropriateplaceinthetextandthepublicationsarelistedhereafter.Datedreferences, subsequentamendmentstoorrevisionsofanyofthesepublicationsapplytothisEuropeanStandardonlywhen incorporatedinitbyamendmentorrevisi
23、on.Forundatedreferencesthelatesteditionofthepublicationreferredtoapplies (includingamendments). EN481, WorkplaceatmospheresSizefractiondefinitionsformeasurementofairborneparticles EN482,WorkplaceatmospheresGeneralrequirementsfortheperformanceofproceduresforthemeasurementof chemicalagents EN689,Workp
24、laceatmospheresGuidancefortheassessmentofexposurebyinhalationtochemicalagentsfor comparisonwithlimitvaluesandmeasurementstrategy EN1232, WorkplaceatmospheresPumpsforpersonalsamplingofchemicalagentsRequirementsandtestmethods EN12919, WorkplaceatmospheresPumpsforthesamplingofchemicalagentswithavolumef
25、lowrateofover5l/min Requirementsandtestmethods ISO7218, MicrobiologyoffoodandanimalfeedingstuffsGeneralrulesformicrobiologicalexaminations 3 Termsanddefinitions ForthepurposesofthisEuropeanStandardthefollowingtermsanddefinitionsapply: 3.1 actinomycetes variedgroupofrodshapedtofilamentousGrampositive
26、bacteria. NOTE Filamentousactinomycetesformabranchingnetworkofthinfilamentscalledamycelium.Mostactinomycetesreplicate byconidialikesporeswhichcaneasilybemadeairborne.Page5 EN13098:2000 3.2 bacteria largegroupofprokaryoticmicroorganismswithonechromosomeinanuclearregionandwhichreplicateonlyasexu allyb
27、ycelldivision. NOTE DifferentcellwallchemistryisusedfortheclassificationofGrampositiveandGramnegativebacteria.Morphological criteriadivideintospheres(cocci)androds.Somespeciesproduceendosporesassurvivalunits. 3.3 bioaerosol airborneparticleswithbiologicalorigin NOTE1 Inthisstandard,microorganismsand
28、endotoxinareconsidered. NOTE2 Themajorityofbioaerosolparticlesarecomplexasfarassize,shapeandconstituentelementsareconcerned 3.4 biologicalpreservationefficiency thecapacityofthesamplertomaintaintheviabilityoftheairbornemicroorganismsduringcollectionandalsotokeep themicrobialproductsintact. 3.5 colon
29、yformingunit theunitbywhichtheculturablenumberisexpressed. NOTE1 Onecolonyformingunitcanoriginatefromonesinglemicroorganism,anaggregateofmanymicroorganismsorfrom oneormanymicroorganismsattachedtooneparticle. NOTE2 Thenumberofoutgrowncoloniesmaydependoncultivationconditions. 3.6 culturablenumber then
30、umberofmicroorganisms,singlecellsoraggregatesabletoformcoloniesonasolidnutrientmedium. NOTE Theviablenumber(see3.21)includesallpotentiallymetabolicactivemicroorganisms.Somemicroorganismsareviable butarenotnecessarilyculturable 3.7 elevatedlevel alevelabovenormalbackgroundlevelofmicroorganismsinaspec
31、ifiedenvironment. 3.8 endotoxin aconstituentoftheexternalmembraneofGramnegativebacteria(lipopolysaccharide),consistingofacomplexlipid, lipidA,whichiscovalentlyboundtoapolysaccharide. NOTE “Freeendotoxin“isliberatedaftercelldeathandbybuddingfromlivingcells.LipidAistheactive(toxic)partandis a potentpr
32、oinflammatorysubstanceandmayinducefebrile,bronchialandothersymptomsinexposedworkers.Thecomposition andthetoxicityofendotoxindiffersbetweenspecies. 3.9 endotoxinunit aunitstandardizedagainstthedefinedreferencematerial(Referencestandardendotoxin). 3.10 exposure(byinhalation) asituationinwhichachemical
33、orbiologicalagentispresentinairwhichisinhaledbyaperson(seeEN482). 3.11 filtration collectionofparticlessuspendedingasorliquidbyflowthroughaporousmedium.Page6 EN13098:2000 3.12 fungi adiversegroupofeukaryoticmicroorganismswithmembraneboundnucleuscomprisingseveralchromosomes. NOTE Multiplicationismain
34、lyasexualbutseveralgroupsreplicatealsobysexualspores.Filamentousfungi(moulds)growin lengthyhyphaeandformcompacttuftscalledmycelia.Asexualspores(conidia)areeasilymadeairborne.Yeastsareusually unicellular,ofsphericalshapeandtheircellsmultiplysexuallyorasexuallybybudding. 3.13 impaction collectionofair
35、borneparticlesacceleratedthroughanozzleororificeonasurfacebyinertiaeffect. 3.14 impingement combinationofimpactionontoasurfaceandsubsequentdispersionintoaliquidmedium. 3.15 LimulusAmoebocyteLysate enzymesextractedfromthebloodcellsofthehorseshoecrab(Limuluspolyphemus)thatareactivatedbyendotoxin. 3.16
36、 microorganisms anymicrobiologicalentity,cellularornoncellular,capableofreplicationoroftransferringgeneticmaterial,orentities thathavelosttheseproperties. NOTE Thetermmicroorganismcoversthetermofbiologicalagent,accordingtotheDirective90/679/EEC:microorganisms, includingthosewhichhavebeengeneticallym
37、odified,cellculturesandhumanendoparasiteswhichmaybeabletoprovokeany infection,allergyortoxicity. 3.17 physicalsamplingefficiency thecapacityofthesamplertocollectparticleswithspecificsizessuspendedinworkplaceair. 3.18 sievesampler multiorificeimpactor. 3.19 totalsamplingefficiency theproductofthephys
38、icalsamplingefficiencyandthebiologicalpreservationefficiency. 3.20 totalnumberofmicroorganisms thenumberofmicroorganismsdeterminedassingleorganisms(oracorrespondingmeasure). NOTE Boththeviableandthenonviablemicroorganismsareincluded. 3.21 viablenumberofmicroorganisms thenumberofmicroorganismshavinga
39、potentialformetabolicactivity. NOTE Aviablemicroorganismisnotnecessarilyculturable,whichmeansthatthenumberofculturablemicroorganismsmay underestimatetheviablenumber. 3.22 workpattern thedefinableseriesofactivitiesfromtheperiodsunderconsideration(seeEN689). 3.23 workplace theworkplaceisthedefinedarea
40、orareasinwhichtheworkactivitiesarecarriedout(seeEN689).Page7 EN13098:2000 4 Symbolsandabbreviations ATP Adenosinetriphosphate CFU Colonyformingunit CV Coefficientofvariation DNA Deoxyribonucleicacid EU Endotoxinunit GSD Geometricstandarddeviation LAL LimulusAmoebocyteLysate LPS Lipopolysaccharide(se
41、e3.8) SEM Scanningelectronmicroscope 5 Measurementofmicroorganismsandendotoxin 5.1 Biologicalagentsandbiologicalproperties Bioaerosolsmaycontaindifferentmicroorganismsand/ordifferentcomponentsoriginatingfromthese.Microorganisms maybeclassifiedindifferenttaxonomicgroupslikeGrampositiveandGramnegative
42、bacteria,actinomycetes,fungi, protozoa,algaeandviruses.Thesemaybefurtherclassifiedtogenusorspecieslevel.Immunologicreactionse.g.allergic and/ortoxicreactionscanresultfromexposuretomicroorganismsirrespectiveoftheirviability. NOTE Speciesidentificationmaybeusefulinstudiesofspecificallergicandtoxicreac
43、tions. 5.2 Aimofmeasurement Themeasurementofmicroorganismsandendotoxinintheworkplaceairhastwoobjectives: toassessworkersexposure;and/or toassessthebiologicalcharacteristicsofairatdifferentlocationsand/oratdifferenttimesandoverdifferent intervalsoftime. Itisessentialtostatethepurposeofthemeasurementa
44、ndhowtheresultswillbeinterpreted. NOTE Measurementtaskscouldbetolocatethesourcesemittingmicroorganisms,tomeasureaworkersdailyorworkshift exposure,toidentifypeaksinexposure,totesttheefficiencyofcontrolmeasures,ortocontrolactionstakentodiminishth e exposure.Itisessentialtoverifythatthesamplingandanalysiscorrespondstotheassessmentobjectivesandtothesearchf or thetargetedcomponentofthebioaerosol(see5.1).Itc
