Animalsarenormallyeuthanizedattheendofastudyforthepurposeofsamplecollectionorpost-mortemexamination.Animalsmaybeeuthanizedbecausetheyareexperiencingpainordistress.Euthanasiaisdefinedasapain-freeorstress-freedeath.TheIACUChasapprovedcertainmethodsforhumanelykillinganimalsthatmeetthedefinitionofeuthanasia.Theappropriatenessofthemethodmayvaryfromspeciestospecies.TheseguidelinesareadaptedfromthereportoftheAmericalVeterinaryMedicalAssociationPanelonEuthanasia,JAmVetMedAssoc2007..TheAmericanCollegeofLaboratoryMedicinehasissuedareportonrodenteuthanasiathathasmanyusefulreferencesandguidelines.

InvestigatorsortechnicianswhorequireadviceorassistanceonpropertechniquesofeuthanasiamaycontactRAR.

NOTE:YoumayonlyuseaeuthanasiamethodthatisapprovedinyourIACUCAnimalCareandUseProtocol.Achangeineuthanasiamethod,includingdoseorrouteofadmiNISTration,isasignificantchangeinprotocolandmustbereviewedandapprovedbytheIACUCbeforeimplementation.

CRITERIAFOREUTHANASIA

Euthanasiaofanimalsisexpectedifanimalsdemonstratetheconditionslistedbelow,whethertheanimalhasbeenmanipulatedornot.AdditionalcriteriamaybespecifiedontheAnimalUsageForm.Fulfillmentofonecriterioncanconstitutegroundsforeuthanasia.ExceptionsarepermittedonlyifapprovedbytheIACUCaspartoftheprotocolreviewprocess(i.e.theclinicalsignslistedbelowareexpectedaspartoftheexperimentandappropriatemeasuresaretakentominimizepainordiscomfortintheanimals).

Weightloss:lossof20-25%(dependingonattitude,weightrecordedattimeofarrival,andage:growinganimalsmaynotloseweight,butmaynotgainnormally)orifnotmeasured,characterizedbycachexiaandmusclewasting.
Inappetance:completeanorexiafor24hoursinsmallrodents,upto5daysinlargeanimals;partialanorexia(lessthan50%ofcaloricrequirement)for3daysinsmallrodents,7daysinlargeanimals.
Weakness/inABIlitytoobtainfeedorwater:Inabilityorextremereluctancetostandwhichpersistsfor24hours,assumingthattheanimalhasrecoveredfromanesthesia.
Moribundstate:depressioncoupledwithbodytemperaturebelow99F,ornon-responsivetostimulation,assumingthattheanimalhasrecoveredfromanesthesia.
Infection:infectioninvolvinganyorgansystem(eitherovert,orindicatedbyincreasedbodytemperatureorWBCparameters)whichfailstorespondtoantibiotictherapywithinanappropriatetimeandisaccompaniedbysystemicsignsofillness.
Signsofsevereorgansystemdysfunctionnon-responsivetotreatment,orwithapoorprognosisasdeterminedbyanRARveterinarian:Respiratory:dyspnea,cyanosis.Cardiovascular:bloodlossoranemiaresultinginhematocritbelow20%;onetransfusionmaybeperformed.Gastrointestinal:severevomitingordiarrhea,obstruction,intussuception;peritonitis,evisceration(immediateeuthanasiarequired).Urogenital:renalfailurecharacterizedbyelevatedBUN,creatinineoruroperitoneum.Nervous:CNSdepression,seizures,paralysisofoneormoreextremities;painunresponsivetoanalgesictherapy.Musculoskeletal:muscledamage,boneinjury,locomotordefecits,etc.resultingininabilitytousethelimb,unlessanticipatedaspartofthestudy.Integumentary:Non-healingwounds,repeatedself-trauma,secondorthirddegreeheatingpadburns.

SURGERYTOCORRECTEXPERIMENTALCOMPLICATIONS

Onlyonemajorsurgicalprocedure(involvingentryofaBDomenorthorax)maybeperformedperanimal,unlessindicatedonanapprovedprotocol.Therefore,majorsurgeryintendedtocorrectcomplicationsarisingafteramajorexperimentalprocedureisnotpermittedwithoutpriorapproval.Insuchcases,euthanasiamustbeperformed.Proceduressuchasrepairofdehiscencesandwoundcleaning/debridementfortreatmentofinfectionmaybeperformedfollowingnotificationoftheRARveterinarystaff.

AcceptableMethodsforEuthanasiaofAnimals

RARformularydosages.

Method	Animalsunder125g	Rabbits/Rodentsover125gunder1kg	Rabbits/Rodentsover1kgunder5kg	Birds	Dogs	Cats	NonhumanPrimates	FarmAnimals(e.g.swine,ruminantsandhorses)	Reptiles	Amphibians³/Fish
CO₂	A	A⁴	A⁴	A	UNA	UNA	UNA	A⁵UNA	A	A
BarbiturateOverdose/EuthanasiaSolution,Intraveneous(100mg/kg)	A	A	A	A	A	A	A	A	A	A
BarbiturateOverdose/EuthanasiaSolution,Intraperitoneal(100mg/kg)	A	A	A	A	UNA	A	UNA	A⁶UNA	A	A
AnesthesiaandExsanguination	A	A	A	A	A	A	A	A	A	A
AnesthesiaandIntraveneousKCl(1-2meq/kg)	A	A	A	A	A	A	A	A	A	A
AnesthesiaandDecapitation	A	A	AWJ	A	UNA	UNA	UNA	UNA	A	A
AnesthesiaandCervicalDislocation	A	A	UNA	A	UNA	UNA	UNA	UNA	N/A	N/A
DecapitationofAwakeAnimal	AWJ¹	AWJ	AWJ	AWJ¹	UNA	UNA	UNA	UNA	AWJ	AWJ
CervicalDislocationofAwakeAnimal	AWJ²	UNA	UNA	AWJ¹	UNA	UNA	UNA	UNA	N/A	N/A
Other								Stunningorelectrocutionfollowedbyexsanguinationmaybeusedforon-farmorslaughterhouseeuthanasia.RifleshotmaybeusedforfieldeuthanasiawhererestraintisnotpossIBLe.		A-ImmersioninMS-222(tricaine)orbenzocaineat2g/Lwater

Volatileagentsusedtoeuthanizeanimalsshouldnotbestoredorusedinanimalroomsbecauseofimproperventilation,toxicitytolaboratoryanimals,andpossibleeffectsonexperimentalresults.

Chloroformisnotacceptableforeitheranesthesiaoreuthanasiaasitisverytoxictomanyspeciesofmice.Additionally,thiscompoundhasbeenshowntobecarcinogenic.

Etherisirritating,flammableandexplosive,andshouldnotbeusedinanimalrooms.Inaddition,animalseuthanizedwithethermustbeleftinafumehoodforseveralhourssothatthecarcassesarenotexplosivewhendisposedof.PrecautionsonetheruseareavailablefromDEHS.

Chloralhydrateandalphachloraloseusedassoleagentsarenotadequatetoreliablyachieveeuthanasia

Abbreviations:

A=Acceptable
AWJ=Acceptibleonlywithscientificjustification,inwriting,ontheAnimalUsageForm,thatanothermethodswouldinterferewiththegoalsoftheexperiment
UNA=Unacceptable
N/A=NotapplicableornotspecificallyaddressedbytheIACUC
AlwaysUNACCEPTABLEinawakeanimals:KCl,MgSO4,strychnine,neuromuscularblockingagents,exsanquination,airembolism,freezingandchloroform(duetoitshazardstopersonnel).

¹⁺²Unlessprecludedbyscientificconsiderations,itisrequiredthatallanimalsbesedatedoranesthetizedbeforedecapitationorcervicaldislocation³Amphibiansmayalsobedouble-pithed⁴ItisrecommendedthatrabbitsnotbeeuthanizedbyCO₂inhalationbecauseofdifficultinduction.⁵Swine<40kgmaybeeuthanizedwithCO2inanappropriatechamber.⁶NeonatalswinemaybeeuthanizedbyIPbarbiturateinjection.

USEOFTHECO₂CHAMBERFOREUTHANASIAOFRODENTS

DIRECTIONS

Wheneverpossible,euthanizeanimalsintheirhomecageratherthantransferringthemtoanewcageorchamberforeuthanasia.
Donotpre-fillthecageorchamberwithCO₂.
Openthetankandadjusttheregulatortoreadnohigherthan5psi.
Slowfillingwillminimizethenasal/occularirritationandaversiontoCO₂.
Waitapproximately3-5minutesforanimaltostopmovingorbreathing.Eyesshouldbefixedanddilated.

THEANIMALISNOTDEADIF:

Itsheartisbeating,checkthisbyfeelingthechestbetweenyourthumbandforefinger.
Itblinkswhenyoutouchtheeyeball.

Iftheanimalisnotdead,placeitbackinthechamber,rechargeandwaitanother5minutesor,usescissorstoopenthechestcavityandcreateapneumothorax.MAKESURETHEANIMALISNOTAWAKEWHENYOUDOTHIS!

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Euthanasia Guidelines

AcceptableMethodsforEuthanasiaofAnimals