Biosafety: Decontamination Methods for Laboratory Use

Decontamination is a term used to describe a process or treatment that renders a medical device, instrument, or environmental surface safe ... 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Decontaminationisatermusedtodescribeaprocessortreatmentthatrendersamedicaldevice,instrument,orenvironmentalsurfacesafetohandle.Sterilization,disinfection,andantisepsisareallformsofdecontamination. Allinfectiousmaterialsandallcontaminatedequipmentorapparatusshouldbedecontaminatedbeforebeingwashed,stored,ordiscarded.Autoclavingisthepreferredmethod.Eachindividualworkingwithbiohazardousmaterialshouldberesponsibleforitsproperhandling. Levelsofdecontamination Theeffectivenessofdecontaminationrangesfromhigh-levelsterilizationtosimplecleaningwithsoapandwater.Levelsofdecontaminationinclude: Sterilizationusesaphysicalorchemicalproceduretodestroyallmicrobiallife,includinghighlyresistantbacterialendospores. Disinfectionusesaliquidchemicaltoeliminatevirtuallyallpathogenicmicroorganisms,withtheexceptionofbacterialspores,onworksurfacesandequipment.Effectivenessisinfluencedbythekindsandnumbersoforganisms,theamountoforganicmatter,theobjecttobedisinfected,andchemicalexposuretime,temperature,andconcentration. Antisepsisistheapplicationofaliquidantimicrobialchemicaltoskinorlivingtissuetoinhibitordestroymicroorganisms.Itincludesswabbinganinjectionsiteonapersonoranimalandhandwashingwithgermicidalsolutions.Manufacturerrecommendationsforappropriateuseofgermicidesshouldalwaysbefollowed. Cleaninguseswater,detergent,andsomemechanicalactionsuchasscrubbingwithaglovedhandorbrush.Cleaningisoftenarequiredstepbeforesterilizationordisinfectionofinanimateobjects becauseitremovesallmaterialsuchassoilororganicmaterialandreducesthenumberofmicroorganismsonanobject. Methods Thereare4maincategoriesofphysicalandchemicalmeansofdecontamination:(1)heat;(2)liquiddisinfection;(3)vaporsandgases;and(4)radiation.Eachcategoryisdiscussedbelow. Heatsterilization(wetordry) Wetheat Wetheatisthemostdependablemethodofsterilization. Autoclaving,sometimescalledsteamsterilization,isthemostconvenientmethodofrapidlyachievingdestructionofallformsofmicrobiallife.Autoclavesusesaturatedsteamunderpressureofapproximately15poundspersquareinchtoachieveachambertemperatureofatleast250°F(121°C)foraprescribedtime—usually30–60minutes. Typicaluses:Autoclavingisadependablemethodofsterilizinglaboratoryequipmentanddecontaminatingbiohazardwastes. Relatedlinks: AutoclaveOverview Autoclave:SterilizeEquipmentandMaterials Autoclave:BiohazardousWaste Dryheat Dryheatislessefficientthanwetheatandrequireslongertimesand/orhighertemperaturestoachievesterilization.Itissuitableforthedestructionofviableorganismsonimpermeablenon-organicsurfacessuchasglass,butitisnotreliableinthepresenceofshallowlayersoforganicorinorganicmaterialswhichmayactasinsulation. Typicaluses:Sterilizationofglasswarebydryheatcanusuallybeaccomplishedat160°–170°Cforperiodsof2to4hours. Precautions:Monitorwetanddryheatsterilizersonaregularbasisusingappropriatebiologicalindicators[sporestrips]. Liquiddisinfection Liquiddisinfectantscanbegenerallyclassifiedashalogens,acids,alkalis,heavymetalsalts,quaternaryammoniumcompounds,phenoliccompounds,aldehydes,ketones,alcohols,andamines. Liquiddisinfectanteffectivenessvarieswiththeorganism,concentration,contacttime,andotherconditionsofuse.Selectonlyliquiddisinfectantsthatareconfirmedtobeeffectiveagainsttheorganism(s)present.Noliquiddisinfectantisequallyusefuloreffectiveunderallconditionsandforallviableagents. Typicaluses:Liquiddisinfectantsareusedforsurfacedecontaminationand,whenusedinsufficientconcentration,asadecontaminateforliquidwastespriortofinaldisposal. Alwaysusechemicaldisinfectantsinaccordancewithlabeledinstructions.Donotusechemicaldisinfectantspasttheexpirationdate. Precautions:Themorechemicallyreactiveacompoundis,themorelikelyitistobetoxicandcorrosive. Consultthe SummaryofDisinfectants (Excel) (PDF) forrecommendeddisinfectants,theiruses,andrequirements.NOTE:Thechartprovidesguidelinesforsurfacedecontamination. Forchemicaldisinfectionofliquidbiohazardouswaste,theonlyuniversity-wideapproveddisinfectantforUCSanDiegoisbleach(1partbleachto9partsliquidwaste,30minutecontacttime,followedbysewering). Ifyourlaboratorywishestoinquireabouttheuseofalternativedisinfectantstoinactivateliquidbiohazardouswaste,pleasesendanemailwiththefollowinginformation.Youwillreceivearesponsewithinfive(5)workingdaysregardingapproval. Materialtobedisinfected Chemicaltobeused Concentrationofchemical Contacttime Disposalmethod(sewering,hazardouswastepickup) Relatedlinks: HowtoDisinfectTissueCultureMediainVacuumFlasks SewerDisposal:WhatCanGoDowntheDrain? Vaporsandgases Vaporsandgases,whenusedinclosedsystemsandundercontrolledconditionsoftemperatureandhumidity,provideexcellentdisinfection.Agentsinthiscategoryincludetheaerosol,vapor,orgasphaseofchlorinedioxide,glutaraldehyde,paraformaldehyde,ethyleneoxide,peraceticacid,andhydrogenperoxide. Typicaluses:Vaporsandgasesareprimarilyusedtodecontaminatebiosafetycabinets,animalrooms,andtheirassociatedsystems,bulkyorstationaryequipmentnotsuitedtoliquiddisinfectants,instrumentsoropticsthatmightbedamagedbyotherdecontaminationmethods,androoms,buildings,andassociatedair-handlingsystems. Caution:Duetotheirhazardousnature,contactEH&SBiosafety,(858)534-5366,forspecialmonitoringrequirementsifthesecompoundswillbeused. Relatedlinks: AutoclaveOverview Radiation(ionizingornon-ionizing) Ionizing Ionizingradiationwilldestroymicroorganisms butisnotapracticaltoolforlaboratoryuse. Non-ionizing TheUV-Cbandofultraviolet(UV)radiationcontainswavelengths(250-270nm,265isoptimum)thateffectivelydestroymostmicroorganismsinairandwaterandonsurfaces.OrganismsmustbedirectlyexposedtotheUVlight;dirt,dust,andshadowscanshieldorganisms,limitingUVlampeffectiveness. Typicaluses:Ultravioletradiationistypicallyusedtoreducelevelsofairbornemicroorganismsandmaintaingoodairhygieneinairlocks,animalholdingareas,ventilatedcabinets,andlaboratoryrooms.UVisalsousedinbiologicalsafetycabinets(BSC)andinsomelaboratoryroomstoreducesurfacecontamination. EH&SBiosafetystronglydiscouragesUVlampsinBSCs.SeeBiosafetyCabinets:UsageGuidelines. Precautions:UVcancauseburnstotheeyes(photokeratitis)andskinofpeopleexposedforevenashortperiodoftime. Usetheseprecautions: ActivateUVlightsonlywhentheareaisunoccupied. UsepropershieldingwhenUVlampsareinuse. InterlockUVlampsusedforspacedecontaminationwiththegeneralroomorcabinetillumination,soturningonthelightsturnsofftheUV. WarnnewandvisitingpersonnelaboutthehazardouseffectsofUVradiationandtheappropriateprecautionstotake. Maintenance BecauseUVlampintensity(itsdestructivepower)decreaseswithtime,propermaintenanceiscriticalfordecontaminationpurposes.Performthisregularmaintenance: ChecklampefficiencymonthlywithaUVmeterormonitoringstrip. CleanUVlampseveryfewweekstopreventdustanddirtfromaccumulatingthatdrasticallyreducesUVlampeffectiveness. Contact EH&SBiosafety. Biosafety Autoclaves BiohazardUseAuthorization BloodbornePathogensProgram ClinicalTrialswithrDNA Containment Decontamination ForTissueCultureMedia Disinfectants Forms GeneDrives GuidelinesforRecombinant/SyntheticNucleicAcidMolecules SelectAgents TissueCultureRooms AnimalBiosafetyLevelReclassification Departments Environment,Health&Safety Seealso Autoclaves BiologicalSafetyCabinets SewerDisposal:WhatCanGoDowntheDrain?