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Introduction: Materials:: Spacerequirements SterilizationofInstruments: Animalpreparation: PreparationoftheSurgeon: PostoperativeCare: ChangesinProcedure: Recordkeeping: 
Asepticsurgeryissurgeryperformedwithoutcontaminationorexposuretopathogens.Thesepoliciesandguidelinesareprovidedtohelpensureoptimalresultsforasepticsurvivalsurgeryusinglaboratoryrodents.Note:Asterileinstrumentpackshouldnotbeusedonmorethanfiveindividualrodents.
SurgicalArea - benchcoatpaperorothercleancoveringcolddisinfectant SterilizationofInstruments - colddisinfectant - cottontippedswabs - guazesquares - waterorsaline - glassbeadsterilizer(repetitivesurgeries) - steriledrapes AnimalPreparation - anesthetic(verifywithinexpirationdate) - antisepticscrub(chlorhexADIneorequiv.) - isopropylalcohol - clippersordepilatorycream - ophthalmicointment(Paralubeorequivalent) - steriledrapes - heatlamp - heatingpad SurgeonPreparation - sterilizedgloves - gownorcleanscrubtop - facemask PostoperativeRecovery - Heatsource:heatlamp,heatingpad,hotwaterbottle(oranytightsealed,flatsurfacedcontainer). - Analgesic(verifyexpirationdate):postoperativeanalgesiamustbegiveninaccordancewiththecorrespondingapprovedYACUCprotocol. - antibiotic:asrequired(verifyexpirationdate) Separatespacesshouldbeusedforthefollowingactivities:1) animalpreparation 2) surgicalareaand 3) postoperativerecovery Acleanspaceshouldbededicatedtosurgeryandmaintainedassuchduringthesurgicalprocedure.Surfacesshouldbewipedwithdisinfectantbeforeandaftersurgery.Thesurgicalareaalsoshouldbecoveredwithacleandrapeorbenchcoatpaper.Instrumentsmustbecleanedpriortopackagingforsterilization.Sterilizationindicatorsmustbeusedinsidethesurgicalpacksandautoclavetapeplacedontheoutsideofthepacktoensuresterilizationhastakenplace.Plasticimplantablesalsomustbesterilizedpriortosurgery.Sterilizationcanbeachievedbyseveralmethods:1) Steam:2500F.,15p.s.i.,for30minutes. 2) Gas-Ethyleneoxide:Exposurerequires8-10hoursinaspecialcABInet(gasautoclave).Plasticimplantablematerialsmustbeaeratedfortwotosevendays,dependingonequipmentrecommendations. 3) Liquid-Immersionin: a) 2%glutaraldehydeplus7.05%phenol(Sporocidin,Cidex,Sonacide)Instrumentsmustbeimmersedfortenhoursina1:10solution.Theymustberinsedinsterilewaterorsalinepriortouse. b) Chlorinedioxide(Clidox)-Instrumentsmustbeimmersedforsixhoursina1:5:1solutionandrequirerinsinginsterilewaterorsalinepriortouse. 4) Physical-Hotbeads:Insertioninabedofhotbeadsfor10-15secondsworkswellforsterilizationofinstrumentsusedinperformingmultiplesurvivalsurgeries.However,thismethodonlysterilizestheinstrumenttips.Theinstrumentsmustbefreeoforganicmaterialpriortocontactingthebeads.Instrumentsalsomustbeallowedtocooltoavoidburningtissue.Sterilizedinstrumentsmustbeplacedonasteriledrapetoreducethechancesofcontamination.Anewsterileinstrumentpackshouldbeusedaftereveryfourtofiveindividualrodents. 1) Animalsshouldbepreppedawayfromthesurgicalarea. 2) Animalsshouldbeproperlyanesthetizedpriortoprepping.Anareatwicethesizeoftheexpectedsurgicalfieldshouldbeprepped. 3) Hairshouldberemovedfromtheincisionsitewithclippersoradepilatorycream.Ifdepilatorycreamisused,theskinmustberinsedwelltoavoiddermatitis.Allloosehairanddebrisshouldberemoved.Tapeworkswelltoensurethatallsmall,loosepiecesofhairareremoved. 4) Anophthalmicointmentshouldbeappliedtopreventdryinganddamagetothecornea. 5) Theskinshouldbecleanedwithadisinfectant.Atwo-stagescrub,firstwithacottonswabsoakedinchlorhexadineorequivalentsolutionfollowedbyacottonswabsoakedinisopropylalcoholworkswellforpreparingthesurgerysite.Thesurgicalsiteshouldbecleanedfromthecenteroutwards.Morethanoneswabshouldbeusedifnecessary.Theswabshouldnotbereturnedtothecenterofthesite. 6) Aftertheskinhasbeenprepared,theanimalshouldbeplacedonawarmsurface.Warminghelpstoreducehypothermiaandquickentherecoverytime.Awatercirculatingheatingpadisveryeffectiveforthispurpose.Acleandrapeshouldbeplacedbetweentheanimalandtheheatingpad.Aheatlampdirectedontheanimalalsoisuseful.Closemonitoringisessentialtoensurethattheanimalwillnotsustainathermalinjuryfromtheheatingpadandheatlamp. 7) Theanimalshouldbedrapedwithsterilecovering.Forsmallrodents,sterilegauzepadscanbeused. Personsperformingrodentsurgeryshouldhaveappropriatetrainingtoavoidcomplications.Goodsurgicaltechniquerequiresasepsis,gentletissuehandling,minimaldissectionoftissue,appropriateuseofinstruments,effectivehemostasisandcorrectuseofsuturematerial.Thesurgeonshouldscrubhandsandarmswithadisinfectantsoapanddonamask,gownandsterilizedgloves.Animalsshouldbekeptwarmanddrytopreventhypothermiauntiltheyarefullyrecovered.Animalscanbewrappedinasmallpieceofclothorgauzesquareandplacedunderaheatsourcetospeedrecovery.Monitoringtheanimal=srecoveryfromanesthesia(breathingpattern,mobility,etc.)andsignsofseepageorbleedingfromtheincisionsiteisstronglyencouraged.Animalsmaybereluctanttoreachuptodrinkaftersurgery,somoisteningnormalfoodandplacingitonthecagebottomwillensureaccesstofoodandwater.Surgicalclipsshouldnormallyberemovedwithin7-10days.TheGuidefortheCareandUseofLaboratoryAnimalsstates"Somecharacteristicsofcommonlaboratory-rodentsurgery,smallerincisionsites,fewerpeopleinthesurgicalteam,manipulationofmultipleanimalsatonesittingandbrieferprocedures,asopposedtosurgeryinlargerspecies,canmakemodificationsinstandardaseptictechniquesnecessaryordesirable."Suchchangesshouldbemadeinamannerthatwillnotcompromisethewell-beingoftheoperatedanimalsandmustbereviewedandapprovedbytheYaleAnimalCareandUseCommittee.Thesurgeonshouldkeepalogbookwiththefollowinginformation:1) Dateofsurgery 2) Animalidentification 3) Typeofprocedure 4) Typeofanestheticincludingdoseandroute 5) Typeofanalgesic,ifgiven,includingdoseandroute 6) Typeofantibiotic,ifgiven,includingdoseandroute(note:hamstersandguineapigsareintolerantofmanyantibiotics) 7) Postoperativemonitoring(time,observations,etc.) 8) Complications 9) IACUCapprovedprotocolnumber 10) Thecagecardshouldalsoindicatethedateofsurgeryandprocedureperformed. References:(availablethoughYARC,#5-4722orVCS,#5-2501)- NationalResearchCouncil(1996),GuidefortheCareandUseofLaboratoryAnimalsChapter3Surgery;Chapter4FacilitiesforAsepticSurgery - Cunliffe-Beamer,T.(1993)ApplyingPrinciplesofAsepticSurgerytoRodents - Borkowski,Gary,DVM,(1995)BasicsofAsepticSurgeryandAnesthesiainRodents(video) QuestionsorcommentsaboutthesepoliciesandguidelinesshouldbeaddressedtotheVeterinaryClinicalServicesat785-2501.Preparedby:MargaretBorelli,SusanMorgenstern,Dr.JamesMacy,VeterinaryClinicalServices,SectionofComparativeMedicine;RandiPrete,LynnPantages-Torok,RegulatoryandSafetyServices,YaleAnimalResourcesCenter.Original:September19,1997RevisedwithApproval:April1,1998,March30,1999