SheilChambersSharpe_2020_AVJ
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Topicalwoundanaesthesia:ef cacytomitigatepigletcastrationpainMLSheil,a* MChambersbandBSharpebObjectiveThereisacriticalneedforsafeandeffectiveanalge-sictreatmentstoaddresspainresultingfromsurgicalhusbandryproceduresinlivestock.Pigletcastrationresultsinacutepain andstresstotheanimal;however,itisperformedgloballyon millionsofpigletsannually,oftenwithoutanyanalgesiawhat-so- ever.Tri-Solfen®(AnimalEthicsPtyLtd,YarraGlen,Victoria,Australia)isacombinationlocalanaestheticandantisepticfor- mulationwhich,appliedtopicallytowounds,hasproveneffec- tive,andisregisteredforusetoalleviatepainassociatedwith castration(andotherwounds)inlambsandcalvesinAustralia andNewZealand.Itisalsoreportedtobeeffectivetoreduce paininpigletsfollowingcastration.DesignThisrandomised,blinded,placebo-controlledstudyexaminedthesafetyandef cacyoftheformulation,adminis-teredviaanadaptedwoundinstillationmethod,tocontrolpainbothduringandfollowingpigletcastration.MethodPigletsreceivedTri-Solfenorplacebo,instilledtothewoundimmediatelyfollowingskinincision.A30swaitperiodwasthenobservedpriortocompletingcastration.Painmitiga- tionwasassessedbygradingnociceptiveresistancemovements andpigletvocalresponseduringcastration,aswellasbygrading responsetomechanicalsensorystimulationofthewound(von
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dbygradingnociceptiveresistancemovements andpigletvocalresponseduringcastration,aswellasbygrading responsetomechanicalsensorystimulationofthewound(von Freyandneedlestick)followingcastration.ResultsTherewasasigni cantreductioninnociceptivemotorandvocalresponseduringcastrationandinresponsetomechanicalsensorywoundstimulationuptoandincluding2hfollowingcastration.Therewerenoadverseevents.ConclusionAdministeredviathismethod,Tri-Solfeniseffec-tivetomitigateacuteperi-operativecastrationpaininpiglets.Keywordsanimalwelfare;castration;localanaesthesia;painmanagement;pigsAbbreviationsAUC,areaunderthedB/timewaveform;NSAID,nonsteroidalanti-in ammatory;NRS,numericalratingscaleAustVetJ2020doi:10.1111/avj.12930Thereisacriticalneedforsafeandeffectiveanalgesictreat-mentstoaddresspainresultingfromsurgicalhusbandryproceduresinlivestock.1Thechallengeistodevelopoptionsthataresafe,practicalandsustainableforon-farmuse.Inviewofthelogisticsinvolved,ideallysuchproducts(albeitveteri-naryprescribed)areabletobefarmerapplied.Pigletcastrationiswelldocumentedtoresultinacutepainandstresstotheanimal.2–5Globally,itisperformedonmillionsofpig-letsannually,inmostcaseswithoutanyanalgesiawhat-so-ever.6Injectedlocalanaesthesiaand/orgeneralanaesthesiamaybeeffec- tivetoalleviatepain;however,thesemethodsmayhavewelfare impactsinandofthemselves(e.g.duetopainofadministration, stressofdoublehandlingorsedation,anddelayedrecovery)7and/ormaybelogisticallyorcostprohibitiveduetotherequirementfor directveterinaryadministration.Nonsteroidalanti-in ammatory(NSAID)medications(suchasmeloxicam)providesomeanalgesia
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stprohibitiveduetotherequirementfor directveterinaryadministration.Nonsteroidalanti-in ammatory(NSAID)medications(suchasmeloxicam)providesomeanalgesia 2–4hfollowingtheprocedure,8butdonotaddressthepainoftheprocedureitselforduringthe rstminutesandhoursfollowingtheprocedure,whenpainisacute.4,7,9–17Theuseoftopicalwoundanaesthesiaisanewandevolving eldthatshowspromiseasanalternativeoradjunctivemeansofdeliver- ingeffectivepainmitigationtofarmanimalsinthesesettings.1Tri-Solfen®isacombinationlocalanaestheticandantisepticformula-tionwhich,appliedtopicallytowounds,hasproveneffectiveandis registeredforusetoalleviatepainresultingfromcastration,andothersurgicalhusbandrywounds,inlambsandcalvesinAustraliaandNewZealand.18–21ItisaviscousliquidformulationcontainingLignocaine5%,Bupivacaine0.5%,Adrenalin1:2000andCetrimide 0.5%.Itisgenerallyappliedtopicallytocoatawoundimmediately postcreation.Designedtoproviderapidonsetwoundanaesthesia withprolongedeffect,itisappliedonce-onlyatthetimeofthe woundcreation,followingwhichanimalsarereturnedtotheirdamstorecoverwithoutfurtherinterference.Speci ctocastration,theproductisappliedimmediatelypostskininci-sion,butimmediatelypriortoclampingand/orcuttingthecordandexcisingthetestes.Thisistoensuregoodcoatingoftheremainingcordtissueaswellasthecutskinedge,whichmaybothotherwisegenerate paininthepostoperativeperiod.Administeredinthisfashion,ithasproveneffectivetoreducepaininpigletsupto4hfollowingcastra- tion.22(Bycontrast,simplysprayinglignocaineontothewoundpos-tcastrationisreportedtobeineffectivetomitigatepain.15)Sutherlandetal16reportedthat‘lyingwithoutcontact’(consideredabehaviouralsignofpaininpiglets)wasreducedinTri-Solfen-
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edtobeineffectivetomitigatepain.15)Sutherlandetal16reportedthat‘lyingwithoutcontact’(consideredabehaviouralsignofpaininpiglets)wasreducedinTri-Solfen- treatedpiglets,ascomparedwithuntreatedpigletsinthe rst180minfollowingcastration.However,therewasnoevidenceofasigni cantimpactonpainduringtheprocedureitself,basedonvocalresponseandcortisollevels.This ndingisnotunexpectedastopicalanaestheticscannotbeexpectedtomitigateproceduralpainifappliedonlyimmediatelypriortotheprocedure.Nevertheless, thequestionariseswhether,ifalongerdwelltimeisprovided,Tri- Solfenmaybeeffectivetomitigatetheacuteproceduralpainthat occurswhensubsequentlyseveringthecords.Appliedtopicallytomucosaltissues,lidocainehasanonsettimebetween20and30s23 *Correspondingauthor.,baysheil@bigpond.net.auaAnimalEthicsPtyLtd,YarraGlen,Victoria,3775,Australia;baysheil@bigpond.net.aubInvetusPtyLtd,Armidale,NewSouthWales,2350,Australia PRODUCTIONANIMALS AUSTRALIA’SPREMIER VETERINARYSCIENCE TEXT PRODUCTIONANIMALS©2020TheAuthors.AustralianVeterinaryJournalpublishedbyJohnWiley&SonsAustralia,LtdonbehalfofAustralianVeterinaryAssociation.AustralianVeterinaryJournalThisisanopenaccessarticleunderthetermsoftheCreativeCommonsAttribution-NonCommercial-NoDerivsLicense,whichpermitsuseanddistributioninanymedium,providedtheoriginalworkisproperlycited,theuseisnon-commercialandnomodificationsoradaptationsaremade.1
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suggestingthatarelativelyshortincreaseinTri-Solfendwelltimemaybeeffectivetoanaesthetisethecords.Thiscouldbehighlybene- cial,asapplyingtractionandseveringthecordsarereportedtobethemostpainfulpartsoftheprocedurebasedonmotorandvocal responses.11,13Thisstudythusexamineswhether,intheon-farmset-ting,topicalwoundanaesthesiawithTri-Solfeniseffectivetoprovideproceduralaswellaspostoperativepainmitigationforpigletsunder-goingcastrationifalongerwaittime(minimum30s)isimplemented betweenapplyingthedoseandseveringthespermaticcords.Wehypothesisedthattopicalwoundanaesthesia(withTri-Solfen)appliedwitha30sdwelltimewouldbeeffectivetomitigatepain bothduringandfollowingsubsequentpigletcastration.Ouraims weretoexamineevidenceofperi-operativepainmitigation.Atpresent,thereisnosinglevalidatedparameterformeasuringpaininpiglets.Itisgenerallyaccepted,however,thatpigletsreacttopainina numberofwaysincluding:physiologically,behaviourallyandthrough resistancemovementsandvocalisation.Anextensiveliteraturereviewwasperformedtoidentifytheoptimalparametersforassessingpaininpigletsduringandfollowingcastration.Thisidenti ed:1Scoringofnociceptivemotor(resistance)movementsduringthe procedure(suchasbyvisualanaloguescaleornumericalrating scale[NRS])consistentlyidenti esamarkedincreaseincastratedpigletsthatmaybeamelioratedbyanaesthetictreatment.24–27Itisthusconsideredtoprovideasoundmethodofdocumentingpain andpainalleviationduringcastration.2Analysisofvocalresponseduringtheprocedureisalsoauseful parameterforpainassessment,albeitthatitisanindirectvariable andconfoundedbypigletvocalresponsetorestraintandhan- dling.Resultsarethereforelesssensitiveandrobust.Themostconsistentresultsareseenwhenpigletsarestudiedinacousticallyseparatedenvironments.13,26–283Directwoundsensorytestingisreportedasasensitiveandrepeat-
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ostconsistentresultsareseenwhenpigletsarestudiedinacousticallyseparatedenvironments.13,26–283Directwoundsensorytestingisreportedasasensitiveandrepeat- ablemeasureofpostoperativewoundpainandpainalleviation withanaesthetictreatmentsinlivestock,18–21andthemethodhasbeenrecentlyvalidatedinpigletspostcastration.22Itisthuscon-sideredtoprovideasoundmethodofdocumentingpainandpain alleviationfollowingtheprocedure.4Behaviouralanalysisisalsoawell-establishedvariablefordocu- mentingpostoperativepain;however,postcastrationabnormalities inpigletbehaviouraresubtleandshort-lived,andresultsareincon- sistentbetweentrials.Themostconsistentresultsarefoundwhen speci c‘pain-associatedbehaviours’areconglomeratedtogiveanover-all‘pain-associatedbehaviour’score,andappeartobegreatestinthe rst30min,andupto3hfollowingcastration.7–10,12,14–165Physiologicalmeasures(suchasanincreaseincortisolorAdreno- corticotrophichormonelevels),despitebeingfrequentlyinvesti- gated,donotprovideareliablemethodofassessingpainorpain alleviationinpigletsundergoingcastration.Theseareindirectmarkersofpain,whichresultfromactivationoftheHypothalamic–Pituitary–Adrenal–axis,withcortisolandadrena-linreleaseandactivationofthesympatheticnervoussystem. Whileusefulinsomesettings,theliteratureestablishesthatthese arepoorindicatorsofpainassociatedwithpiglet castration.4,24,29–31Thesemeasuresareseverelyconfoundedbythesurgicalstressresponse(are exresponseinwhichactivationoftheHypothalamic–Pituitary–Adrenalaxisistriggeredbysurgicalincisionorbleedingregardlessofthepresenceorabsenceofpain). Theymaybefurtherconfoundedbytheadministrationofadrena- linwhichiscommonlyco-administeredwithlocalanaesthetics
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regardlessofthepresenceorabsenceofpain). Theymaybefurtherconfoundedbytheadministrationofadrena- linwhichiscommonlyco-administeredwithlocalanaesthetics whenusedinsurgicalsettings.Basedonthese ndings,nociceptivemotorandvocalresponsestocastrationwerechosenasprimaryandsecondaryef cacyvariablesforassessmentofpainmitigationduringcastration,andnociceptiveresponsetowoundsensorystimulationwaschosenasaprimaryef -cacyvariabletoassesspainmitigationfollowingcastration.Asecondef cacyvariableforpainmitigationfollowingtheprocedure(postop-erativepainrelatedbehaviour)hasalsobeenexamined(andcon- rmed)inalargemulti-centrecomplementary eldtrialwhichisreportedseparately.(Thiswasunabletobeassessedinthesame cohortofpigletsundergoingwoundsensorytesting,asthisinvolvesfrequentre-handling,andhencedisturbanceofpigletbehaviour.)MaterialsandmethodsThestudywasconductedinDecember2018andcompliedwiththefollowingnationalandinternationalstandards:VICHGL9Good ClinicalPractice(issuedJune2000);APVMADataGuidelines–Ef -cacyandtargetanimalsafetygeneralguidelines(Part8,01July 2014),andincompliancewithUniversityofNewEnglandAnimalEthicsCommitteeapprovalno.18-100.StudydesignThestudywasaplacebo-controlledclinicalef cacystudy,usingarandomisedblockeddesignbasedonbodyweightsmeasuredpre-treatment.Theexperimentalunitwastheindividualanimalandthe statisticalanimalwasthetreatmentgroup.Malepiglets(commercial hybrid)werecon rmedsuitableforenrolmentintothestudyiftheywerebetween3and7daysofage,ingoodhealth.Thesewereindivid- uallyidenti ed(uniquelynumberedviaeartags),weighedandranked(heaviesttolightest)andtheheaviestandlightestpigletswere excluded.Theremainingpiglets(n=40,meanweight2.2kg,range
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lynumberedviaeartags),weighedandranked(heaviesttolightest)andtheheaviestandlightestpigletswere excluded.Theremainingpiglets(n=40,meanweight2.2kg,range [1.6–3.0kg])weresequentiallygroupedintoblocksoftwoanimalsandrandomlyallocatedtoGroups1and2fromwithineachblock,suchthateachofGroups1and2contained20malepigletsandhadsimilargroupmeanandrangeofbodyweights,(asassessedandcon- rmedusingStatistix10.0[AnalyticalSoftwareInc.,Tallahassee,FL,USA2010]).OnDay0aftertreatment,animalsnumbered20,26and 41wereremovedfromthestudyduetoherniationandsubsequently replacedwithanimalsnumbered61,62and17,respectively.Thisre- allocationwasstatisticallyassessedcon rmingthatthegroupmeanbodyweightsweresimilaracrosstreatmentgroups.InvestigationalveterinaryproductTri-SolfenBatchNo.:181031–1.Composition:50g/LLignocainehydrochloride,5g/LBupivacainehydrochloride,0.048g/LAdrena-line(asacidtartrate),5g/LCetrimide.PlaceboproductBluefooddyewasaddedto0.9%salinesolution(Baxter,Batch:W47P5,Expiry:April2020)attherateof2.5to250mLsalineAustralianVeterinaryJournal©2020TheAuthors.AustralianVeterinaryJournalpublishedbyJohnWiley&SonsAustralia,LtdonbehalfofAustralianVeterinaryAssociation.2 PRODUCTIONANIMALS PRODUCTIONANIMALS
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(matchingtheinvestigationalveterinaryproductcolourandhenceindistinguishablepostapplication).Investigationalveterinaryproductandplaceboproductswereadmin-isteredviaacustomdesigned1mLTri-Solfenapplicatorwithball- pointtip tted(ProdigyInstrumentsPtyLtd,MountKuring-Gai,NSW,Aust),whichwasveri edpriortouse.Treatmentadministra-tion(totaldose)was:piglets1–<2kg–1mL;piglets>2–4kg–2mL.Studyanimalswerehousedinfarrowingpens(2.1m×2.1m)withtheirdamandlittermates,androutinemanagementpracticeswere followed.Pigletshadconstantsucklingaccesstotheirdamandadlibitumaccesstopotablewater.Onthedayofthestudy,pigletswereremovedfromtheirdamsandcastrationandtreatmentwasper- formedasdetailedinthefollowing.Videoandsoundrecordingwere performedduringpigletcastration,andwoundsensorytestingwas performed1minfollowingtheprocedure.Followingthis,piglets werereturnedtotheirdams.Clinicalobservationsofanimalswereperformedconcurrentwithpostoperativepainassessmentsat1min,thence1,2,4,8,12and24hposttreatment.Methodofcastrationandtreatment:1Restrainpigletgentlybut rmlyinKerblpigletcastrationcradle(AlbertKerblGmbH,Buchbach,Germany);2Whensettled,incisescrotum(includingtunica)withonesingletransverseincisionusingasterilescalpeltoexposeandexterioriseeachtestis;3Immediatelyapplytreatmenttowound(40%totaldosetoeachside)tocoattheexposedspermaticcords;4Wait30s;5Then,removetestesbyseveringthecordsasperroutineprocedure;6Applytheremaining20%ofthetotaldosetocoatthecutskin edgeofthewound.VideorecordingduringcastrationAvideo-camerarecordingdevice(iPhone8–AppleInc.,Cupertino,CA,USA)was xedinpositionabovethecradle,andusedtorecordfromtimeofapplicationofthetopicalwoundtreatmenttoapproxi-mately5sfollowingtheseveranceofthesecondspermaticcord (i.e.completionofthecastrationprocedure).Eachpigletwasclearly
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fthetopicalwoundtreatmenttoapproxi-mately5sfollowingtheseveranceofthesecondspermaticcord (i.e.completionofthecastrationprocedure).Eachpigletwasclearly identi edbyplacinganumberlabelnexttothepigletcradlewithinthecameraview.Thebehaviouralresponsetocastrationwasassessed off-line,byablindedassessor,usinganNRSintensityscaleof0to 2ateachoffourtimepoints:(1)tractionon rsttestis,(2)cutting rstspermaticcord,(3)tractiononsecondtestisand(4)cuttingsec-ondspermaticcord.Thenociceptivemotorresponsewasgradedas: 0=nomotorresponse,1=mildmotorresponse,suchasashort-livedlegextensionorkickingbutnomajorbodyresistancemovementinthecradle,2=markedmotorresponse,suchasprolongedleg movementsand/ormarkedbodyresistancemovementinthecradle. Assessmentthusdocumentedthehandlingofeachteste(traction)and cuttingofeachspermaticcord,givingascore(0–2)foreachevent,andacombinedtotalscore(0–8)forthecastrationprocedure.AudiorecordingduringcastrationSoundwasrecordedwithaZoomH2nHandyRecorder(ZoomNorthAmerica,Hauppauge,NY,USA),mountedonastand50cmfromthesnout.Audiorecordingwastimeanddatestampedtocorre- latewithvideodata.Therecordingperiodcommenced30sfollowing treatmentapplicationtothecastrationwound,whichwasmarked withaverbalcue,and nishedwiththeremovalofthesecondtesticleandsettlingofthepiglet.Sound leswereanalysedoff-linebyasoundconsultantwhowasblindedtopiglettreatment.Foranalysis,sound lesweredownloadedfromtheZoomrecorderandimportedtoProTools®(AvidtechnologyInc.,Burlington,MA,USA).TheywerethenplacedontheProToolstimelineattheirtimestampposi-tionandsynchronisedwithaudiofromthevideorecordingstoallow preciseidenti cationofcuttingpoints.Toallowisolationandcom-parativequanti cationofvocaloutputofpigletsduringtheproce-dure,thetimeofcommencingtraction,andtimeofcordseverance
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cuttingpoints.Toallowisolationandcom-parativequanti cationofvocaloutputofpigletsduringtheproce-dure,thetimeofcommencingtraction,andtimeofcordseverance wereannotatedfromthetimestamp(correlatedwiththevideodata). Screenshotsweregeneratedwiththesametimedurationwindow (xaxis)andsignalscale(yaxis).TheAreaUnderthedB/time(wave-form)Curve(AUC)wasthencalculated(inpixels)usingimageanal- ysissoftware(Image-J®U.S.NationalInstitutesofHealth,Bethesda,MD,USA),foreachofthefollowingperiods:(1)fromcommencingtractionuntilseveranceofthe rstspermaticcord,(2)fromseveranceofthe rstspermaticcorduntilsettledattheendoftheprocedure,and(3)totalrecordedprocedure(Figure1).WoundsensitivitytestingTheresponseofeachpiglettosensorytestingofthewoundsitewasassessedwithavonFrey lament(300g lament)andthencepin-prick(18G,1.5inchneedle)at1min,thence1,2,4,8,12and24h posttreatment,bytrainedstaff,blindtopiglettreatment.Eachassess- mentwasscoredonascaleof0–3,basedonLomaxetal22:0=nomotorresponse;1=therewasamildlocalmotorresponse–includingalocalmuscletwitch, ickofthetailorpuckeringoftheanus;2=therewasapartiallowerbodywithdrawalresponse–includingliftingtherumpfromthecradleand/orprominentmovements(e.g.extensionor escapemovements)oftherearlegs;3=therewasafullbodywith- drawalresponse–includingliftingtherumpfromthecradleandtho-racicmovementsand/orvigorousmovementofboththefrontand rearlegs.Animalswererestrainedsimilarlytothatusedforthecastra-tionprocedureinthepigletcradlefortheseassessments.Pointsofapplicationofthevon-Freyorneedletipforeachsiteincluded:Cut edge;bothlateralaspectsofcastrationwound–2sites.Intactskin:dorsalandventralaspectsofthecastrationwound~3mmfromcut
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eyorneedletipforeachsiteincluded:Cut edge;bothlateralaspectsofcastrationwound–2sites.Intactskin:dorsalandventralaspectsofthecastrationwound~3mmfromcut edge–2sites.Thisgaveamaximumscoreof12foreachtestmethod(vonFreyorneedle)ateachtimepoint.StatisticalmethodsRawdatawereenteredintoMicrosoftEXCEL2016usingdouble-entrytechniques.SummarytableswerepreparedinEXCELwhilesummary gureswerepreparedusingEXCELandTIBCOSpot reS+8.2,2010®(TIBCOsoftwareInc.PaloAlto,CA,USA).Video(nociceptivemotorresponseNRSscores)andAudioAUCdatawerecomparedusingbothparametrict-testsandtheequivalentnonparametrictest(Wilcoxon rank-sumtests);statisticalcomparisonswereperformedusingS+®.Mechanicalsensoryresponsescoredatawerecollatedbytreatment, location(cutedgeandintactskin),timepointandmethodofstimulus (vonFrey lamentsand18-gaugeneedles).TotalscoresforvonFrey,needle-stickandoverallwerecalculated.Scoresto4hposttreatment©2020TheAuthors.AustralianVeterinaryJournalpublishedbyJohnWiley&SonsAustralia,LtdonbehalfofAustralianVeterinaryAssociation.AustralianVeterinaryJournal3 PRODUCTIONANIMALS PRODUCTIONANIMALS
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Figure1.ExamplescreenshotofdB/timewaveformoutputusedforimageanalysisandcalculationof‘areaunderthecurve’(AUC)pigletvocalresponsetocastration.AUCwascalculatedforthefollowingtimeperiods:(1)Commencementoftractiononthe rstspermaticcorduntilsever-anceofthecord,(2)severanceofthe rstspermaticcorduntilsoundreturnedtobaselineattheendoftheprocedure,and(3)totalof1+2. Table1.Groupmeannociceptivemotorresponsescoresfromvideoassessment(numericalratingscale)ofmotorresponsetocastrationproceduresinTri-Solfenorplacebo-treatedpigletsGroupTreatmentTractionon rsttestis(/2)Cut rstspermaticcord(/2)Tractionon2ndtestis(/2)Cutsecondspermaticcord(/2)TotalMotorResponsescore(/8)1Placebo1.52.01.61.86.92Tri-Solfen0.41.40.41.43.7 Figure2.Boxplotoftotalnociceptivemotorresponsetocastration,showingasigni cantreduction(P<0.01)inTri-Solfenversusplacebo-treatedpiglets. Figure3.Boxplotofvocalresponse(areaunderthedB/timewaveformor‘AUC’)duringtimeperiod1(fromcommencingtractionuntilsever-anceofthe rstspermaticcord),showingasigni cantreductioninvocalresponseofTri-Solfenascomparedwithplacebo-treatedpig-lets(P<0.01).AustralianVeterinaryJournal©2020TheAuthors.AustralianVeterinaryJournalpublishedbyJohnWiley&SonsAustralia,LtdonbehalfofAustralianVeterinaryAssociation.4 PRODUCTIONANIMALS PRODUCTIONANIMALS
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werecomparedbytreatment,methodandovertimeusingRepeated-MeasuresAnalysisofVarianceandStatistix10.0®(2013,AnalyticalSoftware,Tallahassee,FL,USA).Suitabilityofthestatisticalmodelwascheckedviasphericityassumptions,assumptionsofcovarianceand residualsplots.Sphericityassumptionsweremetandresidualplots weregenerallyacceptablealthoughassumptionsofcovariancewere generallynotmet.Datawerethereforealsoaggregatedupto‘responsetostimuli/noresponsetostimuli’basedontotalscorecriteria(>2,>3,>4and>5)andtheproportionofpigletsrespondingtostimuliateach timecalculatedto8hposttreatment.Proportionswerethencompared usingChi-SquaredtestsandStatistix10.0.ResultsVideorecordingduringcastrationTri-Solfen-treatedpigletsdemonstratedlowernociceptivemotorresponsescoresassociatedwithtractiononeachtesteandcuttingof eachspermaticcord(Table1).Thetotalmotorresponsescoreandtheresponsestotractionontestes(combined)andcuttingofsper- maticcords(combined)weresigni cantlylowerinTri-Solfen-treatedpiglets(P=0.000;P=0.000;P=0.004,respectively)thanin placebopiglets(Figure2).AudiorecordingduringcastrationTreatmentwithTri-Solfenresultedinasigni cantreductioninvocalresponseofpigletstocastrationascomparedwithplacebotreatmentasmeasuredbyAUC(mean SDof5070 5667versus12,109 9270pxls,respectively)duringthe rstrecordedtimeperiod(fromcommencingtractionuntilseveranceofthe rstsper-maticcord),(P=0.007,Figure3).NumericallylowermeanAUC valueswererecordedinTri-Solfen-treatedpiglets,throughoutthe remainder,orwholeoftheprocedure;however,thesedifferences werenotstatisticallysigni cant,duetowidervariability(Figure4).(Asamplesizeof20pergrouppredictedasigni cant[P<0.05]reductioninphase1AUCwith83%power,versus10%powerfor
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icallysigni cant,duetowidervariability(Figure4).(Asamplesizeof20pergrouppredictedasigni cant[P<0.05]reductioninphase1AUCwith83%power,versus10%powerfor thewholeofprocedureAUCassessment.)WoundsensitivitytestingGroupmeanwoundsensitivityscoresbytestandtimepointarepresentedinTable2andFigure5.Placebo-treatedanimalswere moresensitivetoboththevonFrey lamentandneedleandatboththecutskinedgeandintactskinadjacenttothewound,at1minfol-lowingtreatmentthroughtoandincluding2hposttreatment.Thesedifferenceswerestatisticallysigni cantat1minand1hasshowninTable3.At2hposttreatment,agreaterproportionofplacebo- treatedpigletshadwoundpainresponsescoresgreaterthan 4(P=0.04)orgreaterthan5(P=0.01)thanTri-Solfen-treatedpig- lets.Therewerenosigni cantdifferencesbetweengroupsat4–24hfollowingtreatment.ClinicalobservationsandadverseeffectsIntestinalherniation(inguinalhernia)occurredinthreepigletsrequiringtheirremovalfromthetrialaspreviouslynoted.One placebo-treatedpigletwasfounddead24hposttreatment.Postmor- temexaminationrevealedthistobesecondarytohaemorrhagefrom thetesticulararterystump.Therewerenootheradverseevents. Figure4.Boxplotofvocalresponse(areaunderthedB/timewaveformor‘AUC’)duringtimeperiod2(fromseveranceofthe rstspermaticcorduntiltheendofthecastrationprocedure).Thereiswidevariabilityandresultsarenotsigni cantlydifferentbetweengroups.
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iod2(fromseveranceofthe rstspermaticcorduntiltheendofthecastrationprocedure).Thereiswidevariabilityandresultsarenotsigni cantlydifferentbetweengroups. Table2.GroupmeanresponsescoresfrompigletscastratedwithTri-Solfenorplacebotreatmenttostimulationwithvon-Frey lament(300g)orneedlestick,bytest,siteandtimepointSiteTestGroupTreatment1min1h2h4h8h12h24hCutedgeVonFrey1Placebo0.410.480.851.050.850.451.052Tri-Solfen0.050.000.550.851.350.651.10Needle1Placebo0.911.381.352.902.551.702.632Tri-Solfen0.190.300.902.452.952.002.25IntactskinVonFrey1Placebo0.730.430.501.050.600.600.892Tri-Solfen0.050.100.251.101.200.800.90Needle1Placebo1.641.331.952.602.551.402.422Tri-Solfen0.620.550.952.502.701.902.30©2020TheAuthors.AustralianVeterinaryJournalpublishedbyJohnWiley&SonsAustralia,LtdonbehalfofAustralianVeterinaryAssociation.AustralianVeterinaryJournal5 PRODUCTIONANIMALS PRODUCTIONANIMALS
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DiscussionThedirectapplicationoftopicallocalanaesthesiatowounds,topro-viderapidonsetwoundanaesthesiaandpostoperativepainmitigation,isanewandevolving eld.Itisparticularlyrelevanttolivestockindustries,wherethereisanacuteneedforsafeandeffectiveanalgesicproductsthatmaybeadministeredbyfarmerstoaddresspainassoci- atedwithsurgicalhusbandryproceduresintheiranimals.Theresultsobtainedinthistrialareconsideredtoindicatethatinpiglets3–7daysofageundergoingcastrationinanon-farmsetting,topicalwoundanaesthesiawithTri-Solfen,administeredtothe woundimmediatelypostskinincision,andfollowedbyaminimum 30swaitperiod,effectssigni cantpainmitigationduringthesubse-quentcastrationprocedure,aswellasintheminutesandearlyhours followingtheprocedure.Theapplicationwasabletobeincorporatedintostandardcastrationprocedurewithonlyaminimallyprolongedhandlingtimeof30s perpiglet.Therewerenodrug-relatedadverseeventsreported,con-sistentwithpreviousreports.16,22(Theuseoftwoormorecradles,withstaggeredloadingandpiglettreatment,wouldnegateanyaddi-tionalhandlingtimeforoperators).Inthisstudy,weanalysedpigletnociceptivemotorandvocalisationresponsesasevidenceofcastration-relatedpainandpainmitigation. Theliteratureidenti esareductioninnociceptivemotorresponseasarobustandrepeatablemethodofdocumentingpainalleviationdueto localanaesthesiainpiglets.Pigletcastrationwithoutanaesthesia inducesprotractedviolentstrugglingandescapebehaviourfrompig-lets.24Thismotorresponse,variouslytermed‘escapeattempts’,32‘defencebehaviour’26or‘resistancemovements’,27isusuallyaccompa-niedbyaloudvocalresponseandisattributabletothenociceptivewithdrawalresponsetoacutepaininducedduringtheprocedure. Measuredusingfocalassessment,VisualAnalogueScaleorNRS,stud-
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seandisattributabletothenociceptivewithdrawalresponsetoacutepaininducedduringtheprocedure. Measuredusingfocalassessment,VisualAnalogueScaleorNRS,stud- ieshaveconsistentlyreportedthatpigletmotorresponsetocastrationissigni cantlyincreasedinpigletsundergoingcastrationascomparedwithsham-handledcontrols,32and/orissigni cantlyamelioratedthroughtheuseofgeneralorinjectedlocalanaesthetics,indicativeofsigni cantpainalleviation.25–27Ourresults,con rmingamarkedandhighlysigni cantreductionofnociceptivemotorresponseinpigletstreatedwithlocalanaesthetics(aspresentinTri-Solfen)appliedtopi- callyduringcastration,areconsistentwiththesepreviousreports,and Figure5.Groupmeantotalwoundstimulationresponsescores(von-Frey lament[300g]andneedle-stick)frompigletscastratedwithTri-Solfenorplacebotreatment,bysiteandtimepoint. Table3.Statisticalanalysisofresponsetovon-Frey(300g)andneedle-stickstimulationofthewoundfollowingcastrationinpigletsadminis-teredTri-SolfenorplaceboduringcastrationTotalallP-valueGroup<0.001Time<0.001Group×time0.277Sphericityassumptions√CovarianceassumptionsXGroupTimepointMeanStatisticalcomparison(valueswithdifferentlettersdiffer signi cantly[P<0.05])1.PLACEBO03.79C2.TRISOLFEN00.85D1.PLACEBO13.63C2.TRISOLFEN10.95D1.PLACEBO24.68BC2.TRISOLFEN22.65CD1.PLACEBO47.84A2.TRISOLFEN46.90ABAustralianVeterinaryJournal©2020TheAuthors.AustralianVeterinaryJournalpublishedbyJohnWiley&SonsAustralia,LtdonbehalfofAustralianVeterinaryAssociation.6 PRODUCTIONANIMALS PRODUCTIONANIMALS
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indicatethatsigni cantproceduralpainmitigationisachievedthroughtheuseofTri-Solfenadministeredviathismethod.Pigletvocalisationisalsoausefulindicatorofpain.Althoughpigletscommonlyvocalisewhentheyarehandled,andparticularlywhen restrained,theliteratureshowsthat,duringcastration,pigletsmaysquealmoreoften,moreloudlyand/oratahigherfrequencythanpigletsthatareonlybeingheld.9,11–13,26,33Pullingandseveringofthespermaticcordsleadtothegreatestvocalisationresponse.11,13Pigletscastratedwithoutlocalanaesthesiaproduceahighernumberof screamswithhigherfrequenciescomparedwithpigletscastrated withanaesthesia.13,25–28Acaveat,however,isthatinmostcases,these ndingshavebeenrecordedinroomsacousticallyisolatedfromthefarrowingpensandusingavarietyofdifferentmethods.In mostpiggeries,pigletcastrationisperformedinthefarrowingpens withrapidthrough-puttominimisehandlingtimesandreturnpig- letstothesowasquicklyaspossible.Pigletvocalresponsetoeach partofthehandlingandsurgicalproceduresmayoverlapinthisset- ting,andpigletvocalresponsesmaybeimpactedbyproximitytootherpigletsandthesow,aswellasbeingconfoundedbyextraneousnoise,includingsquealsofnearbypiglets.Ourstudywasdesignedto assessef cacy‘in-realpractice’andwasthusperformedinthefarrowingroom.Themethodofanalysisutilised(areaunderthe dB/timewaveformmeasuredinpixels)providedacomparablemea- sureof‘totalvocalresponse’fromeachpigletovertheselectedtimeperiods.Ourresultidentifyingasigni cantreductioninvocalresponseduringthetimefrom rsttractiononthespermaticcorduntilcordseveranceinTri-Solfen-treatedpigletsisconsistentwith previousreportsindicatingareductioninvocalresponsetocastra-
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rsttractiononthespermaticcorduntilcordseveranceinTri-Solfen-treatedpigletsisconsistentwith previousreportsindicatingareductioninvocalresponsetocastra- tion(andspeci callycordtraction)inlocalanaesthetic-treatedpig-lets,consideredindicativeofsigni cantpainmitigation.Whilewedidnot ndthesamesigni cantdifferencewhencomparingvocalresponsesduringtheremainderorwholeoftheprocedure,thismay re ectwidervariabilityinthedurationofthetotalprocedure,and/orpigletresponsetohandlingoverthelongerperiod,andgreater opportunityforconfounding.Directwoundsensorytestingwasusedtoassessevidenceofpainalleviationfollowingtheprocedure.Directsensorytesting(suchasusingvon-Freyorneedlestick)isalongestablishedandvalidated methodofassessingtheef cacyoflocalanaesthesiaandwoundanalgesiainlaboratoryresearchandclinicalsettings,34includinginpigs.Casteletal35reportedtheresultsofstudiesusingvonFrey la-mentassessmentofnociceptivethresholdtoassessincisionpainin pigsandimpactoflocalanaestheticin ltration,andreportedthatthemethodwasrepeatable,andsensitivetotheeffectsoflocalanaes- theticagents.Modi edtechniqueshavebeendevelopedforuseinlivestockspecies‘inthe eld’.Inthissetting,thenociceptivere exresponsetoneedlestickandvonFreyFilamentapplicationtothe woundhasbeenassessedbygradingthevigourofthenociceptive withdrawalresponseatdifferenttimepointsfollowingtheprocedure.Heightenednociceptivemotorresponsehasbeendocumentedintheminutesandhoursfollowingcastration(andotherhusbandryproce- dures)inlambsandcalves,ascomparedwithsham-handledani- mals,and/orwithpreoperativeassessments).Asigni cantreductioninnociceptiveresponseshasbeendocumentedfollowingTri-Solfen
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,ascomparedwithsham-handledani- mals,and/orwithpreoperativeassessments).Asigni cantreductioninnociceptiveresponseshasbeendocumentedfollowingTri-Solfen applicationtowounds,evidentwithin1–3minofapplication,andpersistingforhoursfollowingprocedures,indicativeofsigni cantwoundanaesthesiaorhypoanaesthesia.Thishasbeenassociated withreducedpostoperativepain-relatedbehaviourintreatedanimals overthesameperiod.18–22Mostrecently,thismethodhasalsobeenvalidatedasamethodofpainassessmentinpigletspostcastration, includingef cacyoflocalanaesthetic(injectedandtopical)adminis-tration.Theauthorsconcluded:‘Thismethodprovidesadirectmea-sureforwoundpainandpresenceorabsenceofanaesthesia’.22OurresultsindicatingrelativehypoanaesthesiainTri-Solfen-treatedpig- letsascomparedwithplacebo-treatedcontrolpigletsfrom1minup toandincluding2hfollowingapplicationareconsistentwiththose ofLomaxetal22andindicativeofsigni cantmitigationofwoundpaininthisperiod.Lomaxetaldocumentedalongerdurationofeffect(4h)thanwasevidentinourstudy(2h).Thedosewasthesameinbothstudies;however,therewasaslightdifferenceincastra- tionprocedureinthatLomaxetalusedthetwo(smaller)incision techniquesversusthesingle(larger)incisiontechniqueutilisedin ourstudy.Itisnotknownifthismayhaveanimpactonpostopera- tivewoundsensitivity,whichmaybeanareaforfuturestudy.Postoperativepain-relatedbehaviourwasunabletobeassessedinthisstudy,aspreviouslydiscussed.Anadditionallinkedmulticentre safetyandef cacytrialisreportedseparately.Wedidnotincludea‘sham’groupinthisstudy.Othershavedem-onstratedmotor,vocalandwoundsensoryresponsesincastrated pigletsversusshamanimalsasdocumentedearlier.Thisstudywas
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a‘sham’groupinthisstudy.Othershavedem-onstratedmotor,vocalandwoundsensoryresponsesincastrated pigletsversusshamanimalsasdocumentedearlier.Thisstudywas designedtoinvestigateameliorationofpaininresponsetonoxiousprocedures,(cordtraction,severing,andsoon)whichwouldnotbeassessableinpigletsthatdidnotundergotheprocedures.Further- more,assessmentswouldnotbeabletobeblinded.Similarly,wehavenotcomparedtheuseoftopicalwoundanaesthesiawithinjectedlocalanaesthesia.Othershavedemonstratedmotor,vocalandwoundsensoryresponsesinpigletscastratedwithandwithoutinjectedlocalanaesthesiaasdocumentedearlier.Theintentofthisstudywastoinvestigatetopicalwoundanaesthesiaasastand-alone treatment(suchasforuseinsituationswhereinjectedanaesthesiaisnotavailable).Asbothmethodsofadministrationresultinasigni cantreductioninvocalandmotorresponses,itmaybeconcludedthatlarge numberswouldberequiredtodetectasigni cantdifferencebetweenthetwotreatmentsforthesevariables.Whilethismaybeofinterest, ultimately,thegreatestpointofdifferencebetweeninjectedlocalanaes-thesiaandtopicalwoundanaesthesia,(suchasviaapplicationofTri- Solfen)stemsfromthedisadvantagesofeachmethodofadministration. Injectedlocalanaesthesiaprovidespreoperativeskinanaesthesia;how-ever,itrequiresveterinarypresence,inducespainofinjectionandmayposerisksduetoinadvertentintravascularadministration.Italso requiresa3–15minwaittimeresultingintheneedfor,andstressof,doublehandlingandseparationfromthesow.Topicalwoundanaes- thesiaavoidstheneedforpainfulinjection,doublehandlingandveteri-
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ltingintheneedfor,andstressof,doublehandlingandseparationfromthesow.Topicalwoundanaes- thesiaavoidstheneedforpainfulinjection,doublehandlingandveteri- narypresence;however,itdoesnotprovideanaesthesiaforskinincision.Nevertheless,skinincision(performedinamatterofmilli-sec-onds),isconsideredtheleastpainfulpartoftheprocedure,and,withtopicalwoundanaesthesiaeffectivewithin30s,painisveryshort-lived.Truecomparatorstudieswouldneedtoexaminetheimpactofthese differencesintotal,(preprocedureandpostprocedure)togainatrueunderstandingofcomparativepigletwelfare.Again,thisisconsideredanareaforfuturestudy.©2020TheAuthors.AustralianVeterinaryJournalpublishedbyJohnWiley&SonsAustralia,LtdonbehalfofAustralianVeterinaryAssociation.AustralianVeterinaryJournal7 PRODUCTIONANIMALS PRODUCTIONANIMALS
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Additionally,thereisthepotentialfortopicalwoundanaesthesiatobeusedaspartofamultimodalapproachtopainmitigationinpig- lets.Synergisticanalgesicef cacyhasbeenreportedwhenTri-SolfeniscombinedwithNSAIDmedicationsinotherspecies,36andtopicalwoundanaesthesiamayprovideenhancedpostoperativeanalgesia whenusedincombinationwithNSAIDsorwithgeneralanaesthesia.Thesearealsoareasforfuturestudy.Itisconcludedthatinpiglets3–7daysofageundergoingcastrationinanon-farmsetting,topicalwoundanaesthesiawithTri-Solfen, administeredimmediatelypostskinincisionfollowedbyaminimum 30swaitperiod,effectshighlysigni cantpainmitigationduringthesubsequentcastrationprocedure,aswellasintheminutesandearly hoursfollowingtheprocedure.Signi cantpaincontrolisthusachievedduringthetime-periodsassociatedwithmaximumpainin pigletsundergoingthisprocedure.AcknowledgmentsTheauthorsthankLeonoraPearson,DipRQA(QualityAssurance) InvetusPtyLtd,Armidale,NSW2350,TimothyPearce,Cert.Audio Eng.(SoundAnalysis),Annandale,Sydney2038andBettaPorkPty Ltd,ValentinePlainsQueensland,Australia.Con ictsofinterestandsourcesoffundingResearchwassponsoredbyAnimalEthicsPtyLtd,andcarriedout byindependentveterinaryresearchcompanyInvetusPtyLtd,toGCPandVICHstandards,toaddressnationalandinternationalvet-erinarymedicinesregulatoryapprovalrequirements.DrSheilisan inventorofTri-Solfen®,aFoundingDirectorandindirectShare-holderofAnimalEthicsPtyLtd.References1.WindsorPA,LomaxS,WhitePProgressinpainmanagementtoimprovesmallruminantfarmwelfare.SmallRuminantRes2016;142:55–57.2.PrunierA,BonneauM,VonBorellEHetal.Areviewofthewelfareconse-quencesofsurgicalcastrationinpigletsandtheevaluationofnon-surgical methods.AnimWelfare2006;15:277.3.vonBorellE,BaumgartnerJ,GiersingMetal.Animalwelfareimplicationsof
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