UBCAnimalCareGuidelines

SOP:ACC2012Tech09
Submittedby:KariJones
Lastdaterevised:Feb2015
Revisedby:KrisAndrews,ShellyMcErlane
Dateapproved:October2012

OralDosing(Gavage)inAdultMiceandRatsSOP

Purpose
ThisStandardOperatingProcedure(SOP)describestheprocedureoforallydosingratsand
mice.Thisprocedurewilladministerthecompoundintothedistalesophaguswhereitwill
thenenterthestomach.ThisSOPfollowstheUBCandCCACguidelinesforacceptableoral
dosevolumesinrodents.
Responsibility
TrainedpersonslistedonanapprovedAnimalCareCommitteeprotocolperformingthe
procedure.
AllanimalusersorallydosingrodentsmusthavesuccessfullycompletedtheUBCAnimal
CareServices(orequivalent)RodentBiologyandHusbandrycourse.
References
CanadianCouncilonAnimalCare(CCAC)guidelines(www.ccac.ca)
AdministrationofSubstancestoLaboratoryAnimals:RoutesofAdministrationandFactors
toConsider(http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3189662/)
Materials
Correctlysizedmetalgavage/feedingneedles(seechartbelow)
Correctlysizedplasticgavage/feedingneedles(seechartbelow,balldiametern/a)
RedRubberFlexibleFeedingTube8fr(Ratsonly)
Appropriatesizedsyringes
Solution/compoundtobeadministered
Optional:towelforwrappingratsifusingflexibleredrubberfeedingtubes

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 Oral Dosing (Gavage) in Mice and Rats SOP

RecommendedGavageNeedleSizeandMaximumVolumeofAdministration*
Species Gauge Length Ball Diameter Volume Example
2.53.8cm Fora25gmmouse,the
Mouse 2420G 1.252.25mm <10ml/kg maximumvolumewouldbe
(1.01.5) 0.25ml

3.810cm Fora250 gmrat,the

Rat 2016G 2.254.0mm <10ml/kg maximumvolumewouldbe
(1.54) 2.5ml

*Greaterthantherecommendedvolumesshouldnotbegivenunlessjustifiedandapprovedon
theAnimalCareProtocolandincreasedmonitoringforcomplicationsimplemented.

PROCEDURE
a.Preparation
1. Weightheanimalandcalculatethemaximumvolumethatcanbeadministered(referto
tableabove).
2. Choosethegavageneedlebestsuitedforyouruse.Therearemetalandflexibleplastic
feedingneedlesinvarioussizesformiceandrats.Metalgavageneedlesaretypicallyeasier
touseinmicesincetheycannotbitethroughthetubebutcaneasilydamagethe
esophagusifthemousestruggles.Straightorcurvedmetalgavageneedlesareavailable
andthechoicedependsonpersonalpreferenceandexperience.Flexibleplasticgavage
needles(miceorrats)orredrubberfeedingtubes(forrats)havelesschanceofdamaging
theesophagusbutanimalscanbitethroughthemandtheyrequiresomepracticetouse
effectively.









Metal Gavage Needles Plastic Gavage Needles Red Rubber Feeding Tube for Rats

3. Priortoperformingtheoralgavageprocedure,measurethedistancefromtheoralcavity
totheendofthexyphoidprocess(caudalpointofthesternum)withthefeeding
needle/tubeontheoutsideoftherestrainedanimal(seephotosbelow).Thiswillbethe
distancetheneedlewillbeinsertedintotheesophagus.Markthisdistanceontheneedle
usingapermanentmarkerorasmallpieceoftape.

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 Oral Dosing (Gavage) in Mice and Rats SOP
NOTE: If using a needle that is too long, there is a greater chance of injuring the animal
by damaging the esophagus. Using a needle that is too short may cause the animal to
aspirate the substance.

Ratmeasuringthedistancefromoralcavitytoxyphoidprocess

Rubber Feeding Tube

tube is marked at
Metal Gavage
the correct distance
Needle showing from mouth to
distance to be xyphoid process
inserted

Line indicates
bottom of rib cage

Xyphoid Process

Mouse measuringthedistancefromoralcavitytoxyphoidprocess

Metal Gavage
Needle showing
distance to be
insterted

Line indicates
bottom of rib cage
Xyphoid Process

4. Prefillthesyringeandgavageneedle/tubewiththecorrectvolumeofthecompoundtobe
administered.Wipetheoutsideoftheneedle/tubetoremoveanyofthecompound
coatingtheoutsideoftheneedle/tube(thisensurescorrectdosingaswellaspreventsthe
animalfromtastingpotentiallybittercompounds).

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 Oral Dosing (Gavage) in Mice and Rats SOP

b.Restraint(seeProperrestraintandinitialplacementofGavageNeedles/tubespictures
below).
5. MICE:Gentlyremovetheanimalfromthecageandfirmlyrestraintheanimalinanupright
position.Getagoodscruffofskinoverthemousesshoulders,sothatthefrontlegsare
extendedouttothesideandtheheadandneckareimmobilized.Ensuretheanimalcan
breathefreely(watchtoseeifthechestismoving).
RATS:Gentlyremovetheanimalfromthecageand,ifusingametalorplasticgavage
needle,restraintheratinanuprightposition,usingeitheravholdorcrossoverto
immobilizetheheadandneck.Ensuretheanimalcanbreathefreely.Ifusingaflexiblered
rubberfeedingtube,theratcanbegentlyrestrainedbyhandorbywrappinginatowel
whiletheyaresittingflatonatableorcounter.
ProperrestraintandinitialplacementofGavageNeedles/tubes




Rat



Gentle restraint of rat for passage of red rubber
VHold restraint of rat for passage of metal feeding tube. The rat could also be wrapped in a
gavage needle. Gavage needle is inserted into towel to make the control of the front legs easier.
the left side of the animals mouth and Gavage tube is inserted into the left side of the
directed along the hard palate of the mouth animals mouth and gently slid down the
to the back of the throat. esophagus to the proper distance.




Mouse Gentle scruff of mouse for passage of the
metal gavage needle. Gavage needle is
inserted into the left side of the animals
mouth and directed along the hard palate of
the mouth to the back of the throat.

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 Oral Dosing (Gavage) in Mice and Rats SOP

c.Gavageprocedure(seeAdvancingtheGavageNeedledowntheEsophaguspicturesbelow).
6. GavageNeedle(MetalorPlastic):Slidetheendofthegavageneedleintotheleftsideof
theanimalsmouthbehindthefrontteethandinfrontofthefirstmolar(thediastema),
alongtheroofoftheanimalsmouthslightlytowardstheanimalsleftside(youmayfeel
theridgesofthehardpalateasyouslidetheneedleback).
Oncethegavageneedleisatthebackofthemouth(theanimalusuallygagsatthis
stage),gentlytilttheheadbacktowardsthespinewithgentlepressurefromthegavage
needle.Thisallowstheesophagustobeinastraightlinetothestomach.Thereshouldbe
noresistancewhenpassingthegavageneedle.Thegavageneedleshouldslidedownthe
esophaguswithgravityalone.Thegavageneedlemayneedtobetwistedclockwiseslightly
asitpassestheepiglottisandintotheesophagus.Passtheneedleintotheesophagusuntil
thepremarkedlinereachesthemouth.
RedRubberFeedingTube(ratsonly):Wraptheratlooselyinatowelorholdtheratona
tableorcountersothatitsfrontfeetcannotgrabholdofthefeedingtube.Slidethe
feedingtubeintotheleftsideoftheratsmouth,overthetonguetothebackofthethroat.
Thetubeshouldeasilyslidedowntheesophagus,astheanimaliseithersittingflatonthe
counterorslightlyelevatedwiththeesophagusinastraightlinewiththestomach.Passthe
needleintotheesophagusuntilthepremarkedlinereachesthemouth.Theratwilloften
trytopullthetubeoutwithitsfrontpaws,sogentlerestraintofthefrontlimbsis
necessary.
NOTE:Ifthereisanyresistanceoriftheanimalstrugglesexcessivelyremovethegavage
needle/tubeimmediatelyandensureyouhavegoodrestraintontheanimalbefore
attemptingtopasstheneedleagain.Donotforcetheneedle/tubedowntheesophagus

AdvancingtheGavageNeedledowntheEsophagus
Rat Mouse




Gavage needle is at the
premeasured distance



Once the metal tube is at the back of the
mouth, gently tip the animals head back
towards its spine so that the head and neck
are in a straight line. This will help the tube
slip down the esophagus with no resistance.
Ensure the animal can breathe freely.

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 Oral Dosing (Gavage) in Mice and Rats SOP
7. Oncethefeedingneedleortubeisatthepremeasureddistance,ensuretheanimalis
breathingnormally.Iftheneedleortubeisinthetrachea,theanimalwillstruggleto
breathe.Iftheanimalisbreathingnormally,injectaverysmalltestdose(~0.05ml).If
thereisnochangeinbreathingeffort,thenslowlyinjectthesolution(over23seconds)to
minimizethefluidcomingbackuptheesophagus.Ifinjectinganoilyorviscoussubstance,
usethesmallestvolumepossibleandinjectmoreslowly(over510seconds)andremove
theneedleslowlysononeofthesubstanceisbroughtbackuptheesophagusintotheback
ofthethroat.Inhalinganyoilysubstanceintothelungswillresultinthedeathofthe
animal.
NOTE:If the animal is not breathing normally, immediately remove the gavage needle
from the esophagus and release the restraint.
8. Whentheentiresubstanceisadministered,removethefeedingneedleslowly,inthe
oppositedirectionfrominsertionandreturntheanimaltoitscage.
9. Monitortheanimalforatleast10minutes,toobserveforpotentialcomplications(see
below).Iftherewereanycomplicationssuchas:obviousaspirationorexcessivestruggling,
monitortheanimaluntilitisactingnormallybeforeleavingitfortheday.

d.CleaninginstructionsforNeedlesandFeedingTubes:
1. Metalgavageneedlesshouldbewipedwithgauzebetweenanimals.Cleanwithsoapy
water,rinsewellandallowtodrybeforestoring.
2. Plasticgavageneedlesareusuallymeanttobesingleuse.Discardaftereachanimal.
3. RedRubberFeedingtubescanbereused.Wipewithgauzebetweenanimals.Rinsewith
waterwhenfinishedusing.Thensoakinsoapywaterfor~20minutes.Rinseagainwith
waterandallowtubestodrybeforestoring.
NOTE: Before using plastic tubes again, assess for any cracks or bite marks in the tube.
Flush with water and if any leaks are seen, discard and do not use. Ensure water flows
easily and there is no debris built up in the tube.

e.Complications
1. Insertionofneedleandadministrationofsubstanceintothetrachea/lungs.
2. Aspirationofsolutionintolungsdueto:needlelengthbeingtooshort,notpremeasuring
toensurethecorrectlengthofneedleisinsertedorregurgitation.
3. Perforationoftheesophagus,tracheaorlungs,duetoimproperrestraintandexcessive
strugglingbytheanimal.
4. Damagetothecardiacsphincter(tissueprotectingopeningintostomach).
5. Damagetotheoralcavity.
6. Esophagitis(inflammationoftheesophagus).
7. Othertraumaticinjuriesrelatedtoimproperanimalrestraint.
8. Death.

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 Oral Dosing (Gavage) in Mice and Rats SOP

ClinicalSignsofcomplications(requiringclosemonitoringandpossiblyeuthanasiaifsignsdo
notresolvewithafewhours):
Respiratorydistress/dyspnea(increasedrespiratoryrateandeffort)
Noisybreathingorclickingwhenbreathing
Paleorblueextremities
Hunchedappearance
Squintedeyes(eyesheldpartiallyclosed)
Piloerection
Bloodatnoseormouth
Swellingofneckorunderfrontlegs(duetoairorfluidescapingfromdamaged
esophagus)
Lossofweightduetoinabilitytoswallow

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