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RESEARCHARTICLE Topicalanaesthesiareducessensitivityofcastrationwoundsinneonatalpiglets SabrinaLomax 1☯ ,CharissaHarris 1☯ ,PeterA.Windsor 1³ ,PeterJ.White 2³ * 1FacultyofScience,SchoolofLifeandEnvironmentalScience,TheUniversityofSydney,Camperdown,NSW,Australia,2FacultyofScience,SchoolofVeterinaryScience,TheUniversityofSydney,Camperdown,NSW,Australia☯Theseauthorscontributedequallytothiswork.³Theseauthorsalsocontributedequallytothiswork.*p.white@sydney.edu.au Abstract TheaimofthisstudywastododeterminetheefficacyofthetopicalanaestheticTri-Solfen®intheameliorationofthepainofcastrationinpiglets.Thetrialwasconductedoverathreedayperiod,andblockedacrosssixlitterswith12pigletstreatedondaysoneandtwo,and16ondaythree.Thepigletswererandomlyallocatedbyweightandlitterto1of4treatmentgroups:(i)shamcastration(SHAM;n=10);(ii)surgicalcastrationwithnoanaestheticinter-vention(CAST;n=10);(iii)surgicalcastrationwithpost-operativetopicalanaesthesia(TRI;n=10);(iv)surgicalcastrationwithapre-operativeintra-testicularlignocainehydrochlorideinjection(LIG;n=10).WoundsensitivitytestinginvolvedvonFreymonofilamentsofweights4gand300g,andan18gaugeneedle,usedtostimulatethewoundandsurroundingskinrespectively,atvariouspre-determinedsites.Neonatalpigletsreceivingtopicalanaesthesia(Tri-Solfen®)sprayintocastrationwoundshadsignificantlylowerwoundsensitivityresponsesforupto4h,comparedtothosecastratedfollowingintra-testicularlignocaineinjectionorthosewithnotreatment.Theuseoftopicalanaestheticissuggestedasapracti-calandaffordablemethodofimprovingpigletwelfareduringcastration. Introduction

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tionorthosewithnotreatment.Theuseoftopicalanaestheticissuggestedasapracti-calandaffordablemethodofimprovingpigletwelfareduringcastration. Introduction Castrationofmalepigletsisaroutinehusbandryprocedurecommonlyperformedwithinthefirstfewdaysoflifeincommercialpiggeriesglobally.Extensiveresearchtoassesstheeffectofcastrationonthewelfareofthepigletsindicatesthatcastrationcausessignificantstress,painanddiscomfortthatcanpersistforupto4days[1±4].Castrationofpigletsisusuallyperformedwithoutanaesthesiaoranalgesia,presumablybecausetheanaesthetictechniquescommonlyusedforcomparablesurgeriesinhumanandveterinarymedicine(generalanaesthesiaorsedation,localanaestheticinfiltrationand/orlocalorregionalnerveblockade)areeithertoocomplex,costlyandtimeconsumingtobepracticaloraffordableforuseon-farm.Anumberofpublicationshavedescribedtheeffectiveuseofanalgesicandanaestheticinterventionstoaddressthepainassociatedwithpiglethus-bandryprocedures[5±9]althoughtheseoptionsmaybeconsideredneitherpracticalnoraffordableforlargescaleproductionsystems. PLOSONE|https://doi.org/10.1371/journal.pone.0187988November15,20171/8 a1111111111a1111111111a1111111111a1111111111 a1111111111

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caleproductionsystems. PLOSONE|https://doi.org/10.1371/journal.pone.0187988November15,20171/8 a1111111111a1111111111a1111111111a1111111111 a1111111111 OPENACCESSCitation:LomaxS,HarrisC,WindsorPA,WhitePJ(2017)Topicalanaesthesiareducessensitivityofcastrationwoundsinneonatalpiglets.PLoSONE12(11):e0187988.https://doi.org/10.1371/journal.pone.0187988Editor:FrancescoStaffieri,UniversityofBari,ITALYReceived:June25,2017Accepted:October30,2017Published:November15,2017Copyright:©2017Lomaxetal.ThisisanopenaccessarticledistributedunderthetermsoftheCreativeCommonsAttributionLicense,whichpermitsunrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited.DataAvailabilityStatement:DataareavailableassupplementaryfileS4.Funding:ThisworkwasfundedbyAustralianPorkLimitedwithtopicalanaestheticformulationsuppliedbyBayerAnimalHealth.Thefunderhadnoroleinthedesign,datacollectionandanalysis,decisiontopublish,orpreparationofthemanuscript.Competinginterests:ThisworkwasfundedbyAustralianPorkLimitedaspartoftheirannualcompetitivegrantsscheme.BayersuppliedthetherapeuticagentTri-Solfenforuseinthis trial.

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TheEuropeanUnion(EU)legislatedthatasofJanuary2012,surgicalcastrationofpigsmustutilizeprolongedlocalanaesthesiaand/oranalgesia,withtheaimofremovingtheprac-ticealtogetherby2018.Whilstcost-benefitanalysesconductedinrelationtothislegislationindicatethatgeneralandlocalanaesthesiastrategiesarebeneficial,inotherjurisdictionssuchasAustraliathecostsoflsuchproceduresareconsideredexcessive,particularlywhereveteri-nariansarerequiredbylawtoadministerrestricteddrugssuchaslignocaine[10].Tri-Solfen1(BayerAnimalHealth,Pymble,Australia)isacommerciallyavailabletopicalanestheticspray-onwounddressingwithhaemostaticandantisepticproperties,registeredinAustraliaforthealleviationofpainofcastrationinsheepandcattle.Itcontainslignocaine(40.6g/L),bupivacaine(4.5g/L),adrenalin(24.8mg/L)andcetrimide(5.0g/L)inagelbase,andhasbeenreportedtobeeffectiveinamelioratingwoundpainandimprovinghealingdur-ingmulesing,castrationandtaildockinginsheepandcastrationincalves[11±14].Duetotheefficacyofthisproductforreducingthepostsurgicalpainassociatedwithcastrationinlambsandcalves,andthesimilaranatomicalnatureofthewoundsinducedinporcinecastration,westudiedtheefficacyofTri-Solfen1intheameliorationofthepainofcastrationinpiglets. Materialsandmethods AllexperimentalprotocolswereapprovedbyTheUniversityofSydneyAnimalEthicsCom-mittee(AECApprovalNo.742). Animals

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castrationinpiglets. Materialsandmethods AllexperimentalprotocolswereapprovedbyTheUniversityofSydneyAnimalEthicsCom-mittee(AECApprovalNo.742). Animals Thetrialwasperformedusing40LandracexLargewhitepigletssourcedfromacommercialherdatTheUniversityofSydneyFarmsnearCobbitty,NSW.Pigletswerefrommixedlittersandaged3±5daysoldwhencastrated.Thepigletswerehousedinfarrowingstallswiththesowandtheirlittermates,beforeanduponcompletionofthetrial.Pigletswereremovedfortheprocedureandsubsequentwoundsensitivitytesting,andretainedinlargecratescontainingasawdustfloorcoverforthe5hourdurationofthestudy,withanoverheadheatlampprovidingwarmth.Thepigletswerereturnedtotheirsowsonconclusionoftheobservationperiod.Thetrialwasconductedoverathreedayperiodusingpigletsfromsixlitterswith12pigletstreatedondaysoneandtwo,and16ondaythree.Oneachday,pigletswereweighedandnum-bered1±12(dayoneanddaytwo)or1±16(daythree)withmarkercrayonforidentification. Treatments

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ysoneandtwo,and16ondaythree.Oneachday,pigletswereweighedandnum-bered1±12(dayoneanddaytwo)or1±16(daythree)withmarkercrayonforidentification. Treatments Thepigletswererandomlyallocatedbyweightandlitterto1of4treatmentgroups:(i)shamcastration(SHAM;n=10);(ii)surgicalcastrationwithnoanaestheticintervention(CAST;n=10);(iii)surgicalcastrationwithpost-operativetopicalanaesthesia(TRI;n=10);(iv)surgi-calcastrationwithapre-operative(5mins)intra-testicularlignocainehydrochloride(1mlpertestis)injection(LIG;n=10).Castrationwasperformedbyasingletrainedtechnician.Pigletswereremovedfromthecrateandrestrainedbytheirhindlimbsinaverticalpositionwiththeheaddown.Thescrotumwasexposedandeachscrotalsacandtunicareceivedasmall(1.5cm)incisionwithasterilescalpeltoexpressthetestes.Theexposedtesteswerethenexcisedfollowingincisionofthespermaticcord.ThewholeprocedureincludingTRItreatment,tooklessthan1minuteperanimal.Sham-castratedpigletswerehandledandrestrainedsimilarlytothecastrates,withthescrotumandtestesmanipulated,butnoincisionmade.PigletsintheTRIgroupreceived1mLofTri-Solfen1spraytoeachscrotalincisionuponexposureofthetestisusinga1mLpipettetoapply0.5mLtothecutskinedgeandspermaticcordpriortoexcision.Thepipetteendwas Topicalanaesthesiaforpigletcastration PLOSONE|https://doi.org/10.1371/journal.pone.0187988November15,20172/8Therearenoothercompetinginterests.ThisdoesnotalterouradherencetoPLOSONEpoliciesonsharingdataand materials.

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theninsertedalongthespermaticcordinsidethewoundtoapplyafurther0.5mLtoensurethatallretractedtissuewascoated(12). Woundsensitivitytesting(WST) WSTinvolvedvonFreymonofilamentsofweights4gand300g,andan18gaugeneedle,usedtostimulatethewoundandsurroundingskinrespectively,atvariouspre-determinedsites.Theseprocedureswereappliedasperpreviousstudies[11,12,14].Animalsweretestedimmediatelypriortotreatmentandthenat1minfollowingtreatment.Furthertestingwasthenconductedatevery30minutesuptofourhoursposttreatmentwithanimalsrestrainedinthesamepositionusedforthecastrationprocedure.Thepresenceorabsenceofskinanaesthesiawasdeterminedbyrecordingtheresponsetoasuperficialskinpinprickwithan18Gneedle[15±17].Woundsensitivitywasmeasuredat5locations,presentingfromlefttoright:leftcutskinedge,leftwoundbody,theskininthemiddleoftheincisions(Middle),rightwoundbodyandrightcutskinedge.SiteandvonFreyweight/needlewererandomizedbetweeneachmeasure-mentandpiglettoavoidconfoundingofresultsInvoluntarymotorresponseswerescoredusinganadjustedcustomizednumericalratingscale[11,12,14].Nociceptiveresponsewasgradedonascaleofseverityfrom0to3,includinglocalresponsesatthewoundsiteandcen-tralresponses:(Score0=nomotorresponse,nodiscerniblemovementorreaction;Score1=mildresponseincludingminorsubcutaneousmuscletwitches,minorvocalization;Score2=moderateresponseincludingpartialwithdrawalreflexorstartleresponsewithminormotormovementofthewholehead;moderatevocalization;Score3=severeresponseinclud-ingfullwithdrawalreflexorfullstartleresponsewithmajorheadjerkandvocalization).Allobservationsweremadebythesameexperiencedobserver. Statisticalanalysis

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ingfullwithdrawalreflexorfullstartleresponsewithmajorheadjerkandvocalization).Allobservationsweremadebythesameexperiencedobserver. Statisticalanalysis Fortheanalysis,bodyweightwascodedintothreecategories(WeightCat):(1)0.9±2.28kg;(2)2.3±3.38kg;and(3)3.3±4.7kg.ScoredatawasanalysedusingordinallogisticregressioninAsREML1(Version16,VSNI,UK).Treatment,time,WeightCat,stimulationtype(Stim)andsitewereincludedasfixedeffects,withpigletandlitterastherandomeffects.ZvaluesweredeterminedusinglogitandSEDarrays,andP-valuescalculated.Dataispresentedascumulativeoddsratioswiththestatisticalprobabilitiesofpigletsineachtreatmentgroupdis-playingaresponsescoreofY=0,1,2and3.Ineffect,theseprobabilitiesaretheaveragesofthescoresfromthe2vonFreyweightsandthe18gneedle.Forallstatisticalcalculations,Pvalues<0.05wereconsideredstatisticallysignificant. ResultsanddiscussionWeight Meanpigletweightwas2.7±0.45kg.Therewasasignificanteffectofweightonpigletresponse(P=0.004).Smallerpiglets(WeightCat1)hadthegreatestprobabilityofmoresevereresponsescores,withthelargerpiglets(WeightCat3)leastlikelytorespond(Fig1). TreatmentxStim TherewasasignificantTreatmentxStiminteraction(P<0.001).Severityofresponseincreasedsignificantlywithweightofstimulationfromthe4gvonFreytothe18Gneedleinalltreatmentgroups(Fig2).Therewasnoeffectoftreatmentonresponseto4gstimulation.TheresponseofTRItreatedpigletsandSHAMpigletsto300gstimulationofthewoundweresimilar,witha Topicalanaesthesiaforpigletcastration PLOSONE|https://doi.org/10.1371/journal.pone.0187988November15,20173/ 8

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significantlygreaterprobabilityofnoresponsethanLIGandCASTpiglets.TRItreatedpigletswereleastlikelytorespondtopin-prickwithan18Gneedle. TreatmentxTime TherewasasignificantTreatmentxTimeinteraction(P<0.001).Therewasageneralincreaseinresponseseverityoverthe4hincastratedpiglets(Fig3).At1minpost-castration,therewasnodifferenceintheresponsetostimulationbetweentreatments.TRItreatedpigletswereleastlikelytorespondtostimulationfrom30minto4hpost-castration,withsimilarresponsestoSHAMpiglets.CASTpigletshadthegreatestproportionofresponsescores;>1atalltimepoints.LIGpigletshadsimilarresponsestoCASTfrom1to4hpost-castration.Localanestheticshavebeenfoundtoreduceacutepainassociatedwithcastrationinpigletswheninjectedintothescrotum[5,7,8]However,theyarerarelyincorporatedintoroutinehusbandryproceduresofcommercialpiggeriesinmanycountries,includingAustralia.Thisstudypresentsnewdataontheefficacyoflocalanaetheticsappliedasatopicalsprayforreduc-ingpost-operativewoundsensitivityfollowingcastrationinpiglets.Applicationoftopicalanaestheticprovidedameliorationofpainupto4hpostprocedureforpigletsundergoing Fig1.Probabilityofpigletsofdifferentweightcategoriesdisplayingresponses(Y;0=noresponse,1=minor,2=moderate,3=severe)tostimulation.Resultscombinetheeffectof4gand300gvonFreymonofilamentsand18gneedleforalltimepoints.a,bWeightgroupsnotsharingacommonletteraresignificantlydifferent(P<0.05).https://doi.org/10.1371/journal.pone.0187988.g001 Topicalanaesthesiaforpigletcastration PLOSONE|https://doi.org/10.1371/journal.pone.0187988November15,20174/ 8

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oi.org/10.1371/journal.pone.0187988.g001 Topicalanaesthesiaforpigletcastration PLOSONE|https://doi.org/10.1371/journal.pone.0187988November15,20174/ 8 surgicalcastrationusingthefarmer-applied,spray-ontopicalanaestheticTri-Solfen1.Thesefindingsmayhavewelfareimplicationsforallpigletsundergoingthisandsimilarsurgicalpro-cedures,particularlyasthesetrialsconfirmedthattopicalanaestheticspraycanbereadilyincorporatedintoroutinefarmmanagementpractices. Fig2.Probabilityofpigletsofdifferenttreatmentgroupsdisplayingresponses(Y;0=noresponse,1=minor,2=moderate,3=severe)tostimulationusing4gand300gvonFreymonofilamentsand18gneedle.Resultscombinetheeffectforalltimepoints.a,bTreatmentgroupsnotsharingacommonletteraresignificantlydifferent(P<0.05).https://doi.org/10.1371/journal.pone.0187988.g002 Fig3.Probabilityofpigletsofdifferenttreatmentsdisplayingresponses(Y;0=noresponse,1=minor,2=moderate,3=severe)tostimulationovera4hourtimeperiod.Resultscombinedtheeffectof4gand300gvonFreymonofilamentsand18gneedleforeachtimepoint.a-cTreatmentgroupsnotsharingacommonletterineachtimepointaresignificantlydifferent(P<0.05).https://doi.org/10.1371/journal.pone.0187988.g003 Topicalanaesthesiaforpigletcastration PLOSONE|https://doi.org/10.1371/journal.pone.0187988November15,20175/ 8

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Topicalanesthesia,appliedduringandimmediatelyaftercastrationhaspreviouslybeenfoundtobepracticalandeffectiveforreducingpost-operativepainassociatedwithsurgicalhusbandryproceduresinlambs[11,12,18,19]andcalves[14,20].Localanaestheticagentsimpedenociception,actingdirectlyonnervetissuetoinhibittheconductionofnerveimpulsesresponsibleforthesensationofpain.Theyareabsorbedthroughmucosalsurfacesanddamagedskin,andcaninducerapidandprofoundlocalanaesthesiawhenappliedtoopenwounds[11,12,21±23].TissuedamageresultingfromcastrationresultsinthelocalizedreleaseofchemicalmediatorsincludingsubstancePandbradykinin,invokinganinflammatoryresponse,withvasodilation,oedemaandincreasednociceptionleadingtohyperalgesia[19,24].Thesemediatorscanhaveaprolongedeffect,leadingtoincreasedsensitizationofneuronstonociceptivesignals(hyperalgesia),andexac-erbatedpaintonoxiousstimuli.LocalanaestheticssuppressbradykininandsubstanceP-mediatedsignaling,withattenuationofcutaneousmicro-vascularflareresponsesindam-agedtissueandreducedinflammation,decreasinghyperalgesiaofthewoundandsurround-ingskin.Localanaesthesiaviainjectionofhasbeenshowntoreducepainaccompaniedbysignifi-cantreductionsinthecortisolresponseofsurgicalcastration[5,7].Ourfindingsindicatethattopicalanaestheticspraycausesasimilarreductioninpain,withCASTpigletsdisplay-ingagreaterresponsetowoundstimulationthanSHAMorTRIpiglets.Thesefindingsareconsistentwithpreviousobservationsofpigletsreceivinglocalanaestheticinfiltrationofthespermaticcordandscrotum[7,25]andwithobservationsofabsentorsignificantlyreducedpainresponsesinlambsandcalvestreatedwithtopicalanaestheticintocastrationwounds[12,14].Inthisstudy,pigletshadthegreatestresponsetoneedlepinprickstimulationofthetissuewithinandsurroundingthewound.Thiswasexpectedasthisstimulusisknowntoinduceadetectablepainresponse[15±17].Thismethodprovedsuccessfulatdetectingthepresenceorabsenceofanaesthesiainthewoundandsurroundingtissuesasshamcastratedpigletsdis-playedaresponsetoneedlestimulationatalltimepoints.Castratedpigletsthatdidnotreceivetopicalanesthetichadsignificantlygreaterpainresponsesfrom2to4hoursfollowingcastration,demonstratingtherecruitmentofthepaincascadeinCASTpigletsnotreceivinganypainmanagement.ItappearslikelythatthepainmanagementfromtheinjectedlignocaineinLIGpigletshaddiminishedbythe2htimepoint,astherewasariseinsensitivityfrom30minto1.5h,andtheresponsesofLIGpigletsdidnotdifferfromCASTpigletsafter2h.Thisistobeexpectedastheligno-caineisreportedtohaveeffectfor1.5±2hrs.Inaddition,itispossiblethattheinjectedlig-nocainehadlittleeffectontheskinandperi-testiculartissue,leavingthemvulnerabletonociception.TRIpigletsdisplayedapparentwoundanaesthesiaupto4h,withsignificantlylowerpainscoresthanCASTandLIGpiglets.TheinclusionoflongactingBupivicaineinthetopicalanaestheticmayexplainthisextendedduration.Thisindicatesthattopicalanaestheticsprayiseffectiveatreducingwoundpainforaprolongedperiodfollowingcastration.Responsescoresofcastratedpigletswerenotsignificantlydifferentimmediatelyfollowingcastration(1min).Itishypothesizedthatthiswasduetotheup-regulationofsensoryreceptorsfollowingtheincision,resultinginareleaseofnaturalpainmediatorsincludingendorphinstomitigateimmediatepainassociatedwiththeincisionandtissueremoval.From30minfollow-ingcastrationTRItreatedpigletshadthelowestpainresponsescores,indicatingeffectiveanaesthesiatowoundstimulationforatleast4h.

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ssueremoval.From30minfollow-ingcastrationTRItreatedpigletshadthelowestpainresponsescores,indicatingeffectiveanaesthesiatowoundstimulationforatleast4h. Topicalanaesthesiaforpigletcastration PLOSONE|https://doi.org/10.1371/journal.pone.0187988November15,20176/ 8

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Conclusions Thesefindingssuggeststhattheglobalpigindustriescurrentlyusingpigletcastration,maybenefitfromconsiderationofthesefindingsastheyofferapossiblemeansofimprovingthewelfareofmalepigletsduringthisroutinebutpainfulhusbandryprocedure. Supportinginformation S1File.Anonymiseddataset.Thisisthedatasetusedforanalysisinthistrial.(XLSX) Acknowledgments TheauthorsacknowledgethecontributionsofMrGregoryMacNamaraforhisassistancedur-ingtrialsandDrEvelynHallforstatisticalsupport. AuthorContributions Conceptualization:SabrinaLomax,PeterA.Windsor.Datacuration:CharissaHarris,PeterJ.White.Fundingacquisition:SabrinaLomax.Methodology:SabrinaLomax,CharissaHarris,PeterA.Windsor,PeterJ.White.Supervision:PeterJ.White.Writing±originaldraft:SabrinaLomax,CharissaHarris.Writing±review&editing:PeterA.Windsor,PeterJ.White. References

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PeterJ.White.Supervision:PeterJ.White.Writing±originaldraft:SabrinaLomax,CharissaHarris.Writing±review&editing:PeterA.Windsor,PeterJ.White. References 1.HayM,VulinA,GeninS,SalesP,PrunierA.Assessmentofpaininducedbycastrationinpiglets:beha-viouralandphysiologicalresponsesoverthesubsequentfivedays.ApplAnimBehavSci.2003;82:201±18.2.MoyaSL,BoyleLA,LynchPB,ArkinsS.Effectofsurgicalcastrationonthebehaviouralandacutephaseresponsesof5-day-oldpiglets.ApplAnimBehavSci.2008;111(1/2):133±45.3.PrunierA,BonneauM,BorellEHv,CinottiS,GunnM,FredriksenB,etal.Areviewofthewelfareconse-quencesofsurgicalcastrationinpigletsandtheevaluationofnon-surgicalmethods.AnimWelf.2006;15(3):277±89.4.SutherlandMA.Welfareimplicationsofinvasivepiglethusbandryprocedures,methodsofalleviationandalternatives:areview.NewZealandVeterinaryJournal.2015;63(1):52±7.https://doi.org/10.1080/00480169.2014.961990PMID:252042035.McGloneJJ,HellmanJM.Localandgeneralanestheticeffectsonbehaviorandperformanceoftwo-andseven-week-oldcastratedanduncastratedpiglets.JAnimSci.1988;66(12):3049±58.PMID:32300676.McGloneJJ,NicholsonRI,HellmanJM,HerzogDN.Thedevelopmentofpaininyoungpigsassociatedwithcastrationandattemptstopreventcastration-inducedbehaviouralchanges.JAnimSci.1993;71(6):1441±6.PMID:83258037.WhiteRG,DeShazerJA,TresslerCJ,BorcherGM,DaveyS,WaningeA,etal.Vocalizationandphysio-logicalresponseofpigsduringcastrationwithorwithoutalocalanesthetic.JAnimSci.1995;73(2):381±6.PMID:76017688.JagginN,GerberS,SchatzmannU.Generalanaesthesia,analgesiaandpainassociatedwiththecas-trationofnewbornpiglets.ActaVeternariaScandinavica.2006;48(Suppl1)(S12).9.SutherlandMA,DavisBL,BrooksTA,McGloneJJ.Physiologyandbehaviorofpigsbeforeandaftercastration:effectsoftwotopicalanesthetics.Animal.2010;4(12):2071±9.https://doi.org/10.1017/S1751731110001291PMID:22445382

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dbehaviorofpigsbeforeandaftercastration:effectsoftwotopicalanesthetics.Animal.2010;4(12):2071±9.https://doi.org/10.1017/S1751731110001291PMID:22445382 Topicalanaesthesiaforpigletcastration PLOSONE|https://doi.org/10.1371/journal.pone.0187988November15,20177/ 8

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rchJournal1999;40(3):111±8.25.TurtonJD.Theeffectofcastrationonmeatproductionandqualityincattle,sheepandpigs.AnimalBreedingAbstracts.1962;30(4):447±55. Topicalanaesthesiaforpigletcastration PLOSONE|https://doi.org/10.1371/journal.pone.0187988November15,20178/ 8