S1466252321000141

chunk 0 · 243 tokens

AnimalHealthResearchReviewscambridge.org/ahrReviewCitethisarticle:BaysingerAetal(2021).Proposedmultidimensionalpainoutcomemethodologytodemonstrateanalgesicdrugefficacyandfacilitatefuturedrugapprovalfor pigletcastration.AnimalHealthResearchReviews22,163–176.https://doi.org/10.1017/S1466252321000141Received:19April2021Revised:30August2021 Accepted:30September2021Firstpublishedonline:3December2021Keywords:Piglet;pain;castration;biomarkers;endpoints;mitigationAuthorforcorrespondence:AngelaBaysinger, E-mail:angela.baysinger@merck.com©TheAuthor(s),2021.PublishedbyCambridgeUniversityPress.ThisisanOpen Accessarticle,distributedunderthetermsoftheCreativeCommonsAttributionlicence(http://creativecommons.org/licenses/by/4.0/),whichpermitsunrestrictedre-use,distribution andreproduction,providedtheoriginalarticleisproperlycited. Proposedmultidimensionalpainoutcomemethodologytodemonstrateanalgesic drugefficacyandfacilitatefuturedrugapprovalforpigletcastrationAngelaBaysinger1

chunk 1 · 437 tokens

d. Proposedmultidimensionalpainoutcomemethodologytodemonstrateanalgesic drugefficacyandfacilitatefuturedrugapprovalforpigletcastrationAngelaBaysinger1 ,SherrieR.Webb2,JenniferBrown3,JohannF.Coetzee4,SaraCrawford5,AshleyDeDecker6,LockeA.Karriker7,MoniquePairis-Garcia8,MhairiA.Sutherland9andAbbieV.Viscardi41MerckAnimalHealth,35500W91stStreet,DeSoto,Kansas66018,USA;2AmericanAssociationofSwineVeterinarians,83026thStreet,Perry,Iowa50220,USA;3PrairieSwineCentre,21058thStreetEast,Saskatoon,Saskatchewan,S7H5N9,Canada;4DepartmentofAnatomyandPhysiology,CollegeofVeterinaryMedicine,ColesHall,KansasStateUniversity,Manhattan,Kansas66506,USA;5NationalPorkBoard,1776NW114thStreet,Clive,Iowa50325,USA;6Smithfield,4134US117,RoseHill,NorthCarolina28458,USA;7DepartmentofVeterinaryDiagnosticandProductionAnimalMedicine,CollegeofVeterinaryMedicine,IowaStateUniversity,Ames,Iowa50011,USA;8NorthCarolinaStateUniversity,CollegeofVeterinaryMedicine,1060WilliamMooreDrive,Raleigh,NorthCarolina27607,USAand9Beef+LambNewZealand,154FeatherstonSt.,Wellington,NewZealandAbstractCastrationofmalepigletsintheUnitedStatesisconductedwithoutanalgesicsbecausenoFoodandDrugAdministration(FDA)approvedproductsarelabeledforpaincontrolin swine.Theabsenceofapprovedproductsisprimarilyduetoawidevariationinhowpain ismeasuredinsucklingpigletsandthelackofvalidatedpain-specificoutcomesindividually indistinctfromotherbiologicalresponses,suchasgeneralstressorinflammationresponses withcortisol.Simplyput,tomeasurepainmitigation,measurementofpainmustbespecific, quantifiable,anddefined.Therefore,giventheneedformitigatingcastrationpain,a consortiumofresearchers,veterinarians,industry,andregulatoryagencieswasformedto identifypotentialanimal-basedoutcomesanddevelopamethodology,basedonthe

chunk 2 · 419 tokens

consortiumofresearchers,veterinarians,industry,andregulatoryagencieswasformedto identifypotentialanimal-basedoutcomesanddevelopamethodology,basedonthe knownscientificresearch,tomeasurepainandtheefficacyofmitigationstrategies.Theout- come-basedmeasuresincludedphysiological,neuroendocrine,behavioral,andproduction parameters.Ultimately,thisconsortiumaimstoprovideavalidatedmultimodalmethodology todemonstrateanalgesicdrugefficacyforpigletcastration.Measurableoutcomeswereselectedbasedonpublishedstudiessuggestingtheirvalidity,reliability,andsensitivityforthedirectorindirectmeasurementofpainassociatedwithsurgicalcastrationinpiglets.Outcomestobeconsideredareobservationofpainbehaviors (i.e.ethogramdefinedbehaviorsandpigletgrimacescale),gaitparametersmeasuredwitha pressuremat,infraredthermographyofskintemperatureofthecraniumandperipheryof theeye,andbloodbiomarkers.Othermeasuresincludebodyweightandmortalityrate.Thisstandardizedmeasurementoftheoutcomevariable’sprimarygoalistofacilitateconsistencyandrigorbydevelopingaresearchmethodologyutilizingendpointsthatare well-definedandreliablymeasurepaininpiglets.Theresultingmethodologywillfacilitate andguidetheevaluationoftheeffectivenessofcomprehensiveanalgesicinterventionsfor 3-to5-day-oldpigletsfollowingsurgicalcastration.IntroductionIntheUnitedStates,surgicalcastrationiscommonlyperformedoncommercialpigproduc-tionfarmswithinthefirst3–5daysoflifetopreventtheaccumulationofboartaintandagon-isticbehaviors(Raultetal.,2011),andtheprocedureistypicallyperformedwithoutadministrationofananalgesicoranesthetic.Currentevidencedemonstratesthatneonates experiencepainand,ifleftuntreated,canresultinpermanentneuroanatomicorbehavioral

chunk 3 · 196 tokens

ationofananalgesicoranesthetic.Currentevidencedemonstratesthatneonates experiencepainand,ifleftuntreated,canresultinpermanentneuroanatomicorbehavioral changes(MellorandGregory,2003;Sneddonetal.,2014).Thus,painmanagementisessentialforyounganimals.Thecastrationofpigletsisrecognizedasasignificantwelfareconcern,andguidelinesfortheuseofanalgesiaandoranesthesiahavebeendevelopedandimplementedintheEU andCanada(NationalFarmAnimalCareCouncil,2014;EuropeanCommission,2017).TheEuropeanCommissionreportedpaininterventionmethodsviaasurveyconductedfrom June2016toOctober2016.Theuseofanesthesiaand/oranalgesiaforpigletcastration foundthemixedapplicationofpainmitigationstrategiesfocusingontheconcernsofanimal https://doi.org/10.1017/S1466252321000141 Published online by Cambridge University Press

chunk 4 · 421 tokens

welfare,economicsustainability,practicalapplicationofthemethod,environmentalimpact,andhumanhealthconcerns. Theapplicationofpainmitigationforpigletcastrationinthe EUrangesfromgaseousorinjectableanesthesia(CO2/O2,ketamine,azaperone,isoflurane),localanesthesia(lidocaine), non-steroidalanti-inflammatorydrugs(flunixin,meloxicam, metamizol),orvariouscombinationsforanesthetic/analgesic effect(EuropeanCommission,2017).However,therewasnoconsensusonthebestmethodforanimalwelfarewiththe practicalityofon-farmeaseofuse.Conversely,USfarmersandveterinariansarecurrentlylimitedinaddressingthischallengeduetothelackofanalgesicoranes- theticdrugsintheUnitedStatesapprovedexplicitlywithanindi- cationforthecontrolofpaininswine.Thelackofon-farm analgesicusemaybeduetoalimitedabilitytomakesolidrecom- mendationsforeffectivepainmanagementstrategiesbyveterinar- ians,theaddedcost,time,andeffortinvolvedwithtraining caretakersandimplementingpainmanagementprotocols on-farm,inadditiontoalackofUSFoodandDrug Administration(FDA)-approvedanalgesicslabeledwithanindi- cationforthecontrolofpainforswine(Raultetal.,2011;Tuyttensetal.,2011;O’Connoretal.,2014).TheUSFoodandDrugAdministration(FDA)hasoversightofapprovalandsafety ofallproductsusedinanimals,includingthoseanimalsusedfor humanconsumption.Pharmaceuticalcompaniesmustusemeth- odstoassessanimalresponsesthatarewell-definedandreliableto demonstrateproducts’efficacyandsafetywhenseekingFDAnewdrugapprovalorlabelamendments.VeterinarianscanprescribeFDA-approvedproductsforextra-labelpurposesunderthe AnimalMedicinalUseClarificationAct(AMDUCA).However, theymusthavereliabledatatodemonstratetheefficacyandsafety offoodproductsderivedfromanimalstreatedwithadrug

chunk 5 · 401 tokens

dicinalUseClarificationAct(AMDUCA).However, theymusthavereliabledatatodemonstratetheefficacyandsafety offoodproductsderivedfromanimalstreatedwithadrug approvedforuseinotherspecies.Aliteraturereviewrevealsalackofconsistentdatarelatedtotheefficacyofpainmitigationproductsprimarilyduetothe lackofuniformtestingmethodologyandprotocols(O’Connoretal.,2016).This,inturn,makesevaluatingtheefficacyofpainmitigationinterventionscomplexandhaspreventedconsensus onbestpracticesforpainrelief(Bateson,1991).Lackofconsistentprotocolscreatesdifficultyforpharmaceuticalcompaniestosub- mitnewproductapprovalsorlabelclaimsrelatedtopain,veter- inarianstoconfidentlyprescribeproductsforextra-labeluse, researcherstoreliablyassesspainandpotentialmitigationstrat- egies,andpigfarmerstomakefuturebusinessdecisionsregarding animalwelfare.Giventheneedformitigatingcastrationpain,aconsortiumofresearchers,veterinarians,industry,andregulatoryagencieswas formedtoidentifypotentialanimal-basedoutcomesanddevelop amethodologybasedontheknownscientificresearch,tomeasure painandtheefficacyofmitigationstrategies.Theconsortium’sgoalistoimprovepigwelfareon-farmbyeffectivelycontrolling painassociatedwithon-farmsurgicalprocedures,suchascastra- tion,inamannerthatissafefortheanimalandtheconsumerand iscompliantwithUSregulation.Thisevaluation’sprimarygoalistofacilitateconsistencyandrigorbydevelopingaresearchmeth- odologyutilizingvalidatedendpointsthatarewell-definedand reliablymeasurepaininpiglets.Theresultingmethodology, withvalidatedoutcomes,willfacilitateandguidetheevaluation oftheeffectivenessofcomprehensiveanalgesicinterventionsfor

chunk 6 · 443 tokens

npiglets.Theresultingmethodology, withvalidatedoutcomes,willfacilitateandguidetheevaluation oftheeffectivenessofcomprehensiveanalgesicinterventionsfor 3-to5-day-oldpigletsfollowingsurgicalcastration.Measurableoutcomeswereselectedbasedonpreviousstudiessuggestingtheirvalidity,reliability,andsensitivityforthedirectorindirectmeasurementofpainassociatedwithsurgicalcastrationinpiglets.Outcomestobeconsideredareobservationofpain behaviors(i.e.ethogramdefinedbehaviorsandpigletgrimace scale),gaitparametersmeasuredwithapressuremat,infrared thermography(IRT)ofskintemperatureofthecraniumandper- ipheryoftheeye,andbloodbiomarkers.Othermeasuresinclude bodyweightandmortalityrate.Theinformationhereinsupportstheinclusionofmultipleendpointstoevaluatetheirvalidityandreliabilityfordemonstrat- ingcontrolofpaininpigletsundergoingsurgicalcastration.For endpointmeasuresnotincludedforevaluation,theconsortium’sdecisionwasbasedonalackofvalidatedprocessesorpracticality ofstandardizingtheon-farmapplicationtojustifyusewithinthe proposedmethodology[i.e.vocalizationandnociceptivewith- drawalresponse(SheilandPolkinghorne,2020)].Theseend-pointsmayultimatelybeusedinstudiestodemonstrate substantialevidenceofeffectiveness,onecomponentintheUS FoodandDrugAdministration’sapprovalprocessofapainmiti-gationdrug.Thispaperaimstodescribeamultidimensional methodologytodirectlyorindirectlyassessbehavioral,physio- logical,andneuroendocrinechangesinpigletsassociatedwith painresultingfromsurgicalcastration.Thismethodologywill usemultipleoutcomevariablesto,insummation,demonstrate analgesicefficacyinthepost-surgicallycastratedpiglet,satisfying theFDAefficacyrequirementofaproduct.PaindefinitionPainisdefinedasanunpleasantsensoryandemotionalexperi-enceassociatedwithactualorpotentialtissuedamage(IASP

chunk 7 · 441 tokens

Aefficacyrequirementofaproduct.PaindefinitionPainisdefinedasanunpleasantsensoryandemotionalexperi-enceassociatedwithactualorpotentialtissuedamage(IASP SubcommitteeonTaxonomy,1979).Theemotionalcomponentofpainisanaffectivestatethatcannotbedirectlymeasured. Thesensorycomponentofthepainresponseinvolvesnocicep- tion,includingthedetection,transduction,andtransmissionof noxiousstimulibytheperipheralandcentralnervoussystems. Collectivelythesensorycomponentofthepainresponseproduces behavioral,physiological,andneuroendocrineresponses.EvidenceofpainassociatedwithcastrationHumanbeingsandothervertebratemammals,suchaspigs,havesimilarneuroanatomicalstructuresassociatedwithpain perception[e.g.nociceptors,apathwayconnectingnociceptors tothebrain,andbrainstructurestoprocesspainanalogousto thehumancerebralcortex(Bateson,1991)]andthecapacityforanimalstoexperiencepainiswell-described.Itwaslongbelieved thatneonateswereincapableofexperiencingpainordidsoless intenselythanadultsbecauseoftheirimmaturenervoussystem andlackofspecificbehavioralsigns(Bateson,1991).Formanyyears,thisconceptwastranslatedtoveterinarymedicineandlive- stockproductionpracticesasproducersandveterinariansprovided littletonoanalgesicsoranestheticstoanimalsifpainfulproce- dureswereconductedatayoungage.Evidencenowsuggestsneo- natesmayhaveaheightenedpainexperience,anduntreatedpain couldresultinpermanentchangestopainsensitivityandneuroa- natomicorbehavioralabnormalities,makingpharmaceuticalpain managementevenmorecriticalforyounganimalsundergoinga surgicalprocedure(MellorandGregory,2003;Sneddonetal.,2014).Surgicalcastrationofpigletscausesacutepain,asevidencedbybehaviorandphysiologicchanges.Pigletsdisplayseveralpain behaviorspost-operativelyinresponsetothesurgicalcastration164AngelaBaysingeretal.

chunk 8 · 60 tokens

asevidencedbybehaviorandphysiologicchanges.Pigletsdisplayseveralpain behaviorspost-operativelyinresponsetothesurgicalcastration164AngelaBaysingeretal. https://doi.org/10.1017/S1466252321000141 Published online by Cambridge University Press

chunk 9 · 448 tokens

procedure,includinganincreaseinstiffness,trembling,scratchingtherump,tailwagging,awakeinactiveorrestlessbehaviors,and spendingmoretimeisolatedfromlittermates(Viscardiand Turner,2018a).Pigletsalsoproducedistincthigh-frequencyvoca-lizationsassociatedwiththecastrationprocedure(Wearyetal.,1998;Leidigetal.,2009)andspendmoretimeincontactwiththesow,whichhasbeensuggestedtoproduceanalgesic-likeeffects bypromotingendorphinreleaseinneonates(FieldandGoldson, 1984;Blass,1994).Behavioralalterationsassociatedwithcastrationpersistbeyond24h,withsomeabnormalbehaviorspresent4days later(Hayetal.,2003;LlamasMoyaetal.,2008).Surgicalcastrationalsocausesaphysiologicresponse.Pigletsshowanincreaseinheartandrespirationratewithhigher bloodcortisol,lactate,andadrenocorticotropinhormoneconcen- trationsaftercastration(Whiteetal.,1995;Prunieretal.,2006;Kluivers-Poodtetal.,2012).Peripheralvasoconstrictioncausedbyactivationofthesympatheticnervoussystemresultsina decreaseincutaneoustemperature,which,usinganIRTcamera, hasbeenobservedinpigletsaftercastration(Batesetal.,2014).Surgicalcastrationcanalsoresultinadecreasedgrowthratelead- ingtoproductionlosses(McGloneetal.,1993;Kiellyetal.,1999;Malavasietal.,2006).Publishedstudiesthathaveexaminedanal-gesiaandlocalanestheticuseinpigletsundergoingsurgicalcas- trationaresummarizedinTable1.DevelopmentofaresearchprotocolTheinformationpresentedhereinservesasasupportfordevelop-ingaresearchprotocolwiththeprimaryobjectiveofdetermining thevalidityandreliabilityofendpointsforuseindemonstrating efficacyforFDAproductapproval.Theendpointscouldbe usedtoevaluateadrug’sefficacyforcontrollingpainassociatedwithcastrationof3-to5-day-oldpiglets.The3-to-5day-oldpiglet istargetedduetothestandardageatwhichpigletsarecastratedin commercialproductionwithintheUnitedStatesbeingapproxi-

chunk 10 · 385 tokens

day-oldpiglets.The3-to-5day-oldpiglet istargetedduetothestandardageatwhichpigletsarecastratedin commercialproductionwithintheUnitedStatesbeingapproxi- mately3daysofage,andfoundationalresearchinpainmitigation hastargetedthe3-day-oldpiglet(Sutherlandetal.,2010;Sutherlandetal.,2012).Throughareviewoftheliteraturedescribingpainexpressionmeasuresinpiglets,multipleoutcome measureswereidentifiedasthebestcandidatesforfurthervalid- ationandinclusioninfutureefficacytrials.Thispaper’smultidi-mensionaloutcomesincludeobservationandscoringofpain exhibitionorbehavior,physiologicalbiomarkers,andautomated physicalmeasurements.ProtocolstudydesignInthedevelopmentofastandardizedprotocoltodetermineanal-gesicefficacy,thefollowingprotocolwasoutlinedtoprovidecon- sistencyintrialdesignandstandardizethemeasurementof outcomes:(a)Fourprimarytreatmentgroupswillbeusedintheresearchprotocol:(1)shamcastration(SHAM),(2)shamcastration withtheintervention(SHAM+TRT),(3)surgicalcastration withplacebocontrol(CAST),(4)andsurgicalcastrationwith intervention(CAST+TRT).Intheabsenceofothercon- foundingfactors,fourtreatmentgroupswillhelpdetermine thefollowing(Wearyetal.,2006;Isonetal.,2016): DifferencesbetweenCASTandCAST+TRTtreatmentscanbeinterpretedastheefficacyoftheinterventionto mitigatepain. DifferencesbetweenSHAMandCASTcanbeinterpretedastheeffectsofpainortissuedamageratherthanenviron-mentalorothernon-painfactors. DifferencesbetweenSHAMandSHAM+TRTcandeter-mineiftheinterventioncausesadifferenceinthetested variableintheabsenceofpainandtissuedamage(e.g.seda-

chunk 11 · 419 tokens

ctors. DifferencesbetweenSHAMandSHAM+TRTcandeter-mineiftheinterventioncausesadifferenceinthetested variableintheabsenceofpainandtissuedamage(e.g.seda- tiveeffects).(b)Tovalidatethemeasurableoutcomesinthisprotocol,theauthorswouldusebuprenorphine(0.04mgkg−1)(ViscardiandTurner,2018a),administeredintramuscularly,beforesur-gicalcastrationasa‘goldstandard’intervention.Whilethisopioiddrugisnotapprovedforuseinswineorotherfood animals,buprenorphinewaschosenasthegoldstandard becauseofitspotencyasananalgesicdrugandabilityto bindtoopioidreceptorsinthebrain,spinalcord,andperiph- erytosuppresspainsignaltransmission(Chahl,1996).Furthermore,buprenorphinehasproventobeeffectiveat reducingpainandlamenessinpiglets(Hermansenetal.,1986;Meijeretal.,2015)andalleviatingbehavioralpainindi-catorsandfacialgrimacingwithnosedativeeffectformore than24hin5-day-oldpigletsfollowingcastration(Viscardi andTurner,2018a).(c)Theresearchprotocolwouldusea2×2factorialdesignwithpiglet(withinlitter)astheexperimentalunit.Thetreatment willbeappliedtotheindividualmalepiglet,andeachtreat- mentwillberepresentedwithinalitteratleastonce.If morethanfourmalepigletsinthelittermeettheenrollmentcriteria,atreatmentwillbeassignedtoeachadditionalpigletusingatreatmentrandomizationlist.Usingpigletasthe experimentalunitandblocking/controllingallocationto treatmentbasedonthelitter(controllingforsoweffect) helpscontrolinter-littervariability(e.g.littersize,sowmilk yield,andpigletsexratio)(Festing,2006;LazicandEssioux,2013).Manypreviousstudies(Table1)evaluatingpainreductionforcastratedpigletshaveusedpigletsasthe experimentalunit.Astheexperimentalunit,thepigletpro- videsmoredatafor,andhigherconfidenceinthepowerana-

chunk 12 · 339 tokens

ductionforcastratedpigletshaveusedpigletsasthe experimentalunit.Astheexperimentalunit,thepigletpro- videsmoredatafor,andhigherconfidenceinthepowerana- lysiscalculationsusedtodeterminetheresearchprotocol’ssamplesize.Apotentialcriticismforusingpigletastheexperimentalunitratherthanthelitteristhathavingallfourtreatmentsrepresented withinalittercouldhaveaconfoundinginfluenceonpiglet behaviorandactivity.Emotionalcontagionhasbeenobserved ingroupsofpigswhenexposedtopositiveornegativetreatments (Reimertetal.,2017;Yunetal.,2019).Forthisconcerntobeaddressed,usinglitterastheexperimentalunitintheresearch protocol,alluncastratedmaleandfemalelittermateswould needtoberemovedfromthefarrowingboxsothatonlytreated malesremained,creatinganunnaturalenvironment.Having uncastratedfemalelittermatespresentintheenvironmentis importantwhenvalidatingoutcomesandevaluatingintervention efficacy,astheseanimalswillalwaysbepresentintypicalcom- mercialproduction.MeasurableoutcomesValidity,reliability,andsensitivityarevitalcharacteristicsthatshouldbeconsideredwhenchoosingmeasurableoutcomesfor practicalpainassessment(Isonetal.,2016).Eachmeasurableout-comeidentifiedforuseintheproposedprotocolhasbeencate- gorizedaseitherprimaryorancillary.PrimaryoutcomeswereAnimalHealthResearchReviews165 https://doi.org/10.1017/S1466252321000141 Published online by Cambridge University Press

chunk 13 · 428 tokens

Table1.ResultsofstudiesthathaveexaminedanalgesiaandlocalanestheticusetoalleviatepaininsurgicallycastratedpigletsReferencePigletageAnalgesiaLocalanesthesiaMeasuredoutcomeDrugefficacyaMcGloneandHellman(1988)2weekand7week–2%lidocainehydrochlorideintratesticularNursingbehaviorYes(2weekpigs);No(7weekpigs)Maintenancebehavior(eating,drinking,standing,lying)NoMcGloneetal.(1993)8week22mgkg−1aspirinoralor0.11mgkg−1butorphanolIV–PainbehaviorNoWhiteetal.(1995)1–24d–2%lidocainehydrochlorideintratesticularVocalizationYesHeartrateYesHornetal.(1999)10–14d–2%lidocainehydrochlorideintratesticularVocalizationNoResistantmovementsYesHagaandRanheim(2005)22d–2%lidocainehydrochlorideintratesticularandintrafunicularMeanarterialpressureYesPulserateYes EEGYesZankletal.(2007)4–6d–Procainehydrochloride2%lidocainehydrochloride intratesticularCortisolNoLeidigetal.(2009)3–4d–10mgprocaineintratesticularPainbehaviorYesKeitaetal.(2010)<1week0.4mgkg−1meloxicamIM–PainbehaviorYesCortisolYesSutherlandetal.(2010)3d–CetacaineorTri-solfentopicalVocalizationsNoCortisolNo LeukocytecountNo LyingbehaviorNoHanssonetal.(2011)3–4d1.0mgmeloxicamIM10mgmL−1lidocaine+5 gmL−1epinephrineintratesticularPainbehaviorYes(Monly)VocalizationYes(Lonly)EartemperatureYes(LonlyandM+L)Schmidtetal.(2012)5–7d0.4mgkg−1MeloxicamIM–NursingbehaviorYesActivebehaviorNoSutherlandetal.(2012)3d2.2mgkg−1flunixinmeglumineIM–CortisolNoC-ReactiveProteinNo SubstancePNo VocalizationNoPainbehaviorNoActivityNoKluivers-Poodtetal.(2013)2–5d0.4mgkg−1meloxicamIM2%lidocaineintratesticularPainbehaviorYes(MonlyandM+L)Batesetal.(2014)5dmeloxicamtransmammary–CortisolYesPeripheralvasoconstrictionYesViscardietal.(2017)5d0.4mgkg−1meloxicamIMEMLAtopicalcream(lidocaine2.5%,prilocaine

chunk 14 · 81 tokens

014)5dmeloxicamtransmammary–CortisolYesPeripheralvasoconstrictionYesViscardietal.(2017)5d0.4mgkg−1meloxicamIMEMLAtopicalcream(lidocaine2.5%,prilocaine 2.5%)PainbehaviorNoActivitylevelNoFacialgrimacingNo(Continued)166AngelaBaysingeretal. https://doi.org/10.1017/S1466252321000141 Published online by Cambridge University Press

chunk 15 · 434 tokens

definedasmeasuresdirectlyrelatedtoclinicalsignsofpainasrecordedinthepublishedliteratureandarerepeatablewhenthe propermethodologyisused.Ancillaryoutcomesweredefined asnewermethodologiesinthepublishedliteraturedirectlyrelated toclinicalsignsofpainorindirectlyrelatedtoclinicalsignsof painandsupporttheprimaryoutcomes.Acombinationofmul- tipleoutcomevariablesmayprovidearobustevaluationofa castratedpiglet’spainprofileandthetestedinterventionefficacy.Abriefjustificationisprovidedforeachmeasurableoutcomefortheproposedprotocol.Additionally,evidenceforwhenand howeachoutcomeshouldbemeasuredisprovided.Arecom- mendedsamplesizeisalsoincludedforeachoutcomebasedon previousliterature.Withthediversityinthemethodologyusedintheexistingliterature,performingacompletepoweranalysisproveddifficult.OutcomesvariablesPainbehaviorandactivitytrackingDiagnosisofpaininanimalsisacomplicatedprocessdueto uniqueindividualexperienceswithpain(GaynorandMuir, 2009)anddifferencesinpaintoleranceandreactionbetweenbreeds,sex,age,painduration,proceduretype,andstimulus severity(Matthew,2000).Fromascientificstandpoint,abehav-ioralassessmentmaybethemostpracticalendpointtoassess paininlivestockproductionsystemsasitisthemostdirectassess- mentofananimal’swelfare.However,giventhatthesensorycomponentofpainisalsoassociatedwithaneuroendocrine andphysiologicalresponseintheanimal,measuringneuroendo- crineandphysiologicaloutcomesisessentialtoconfirmandsup- portbehavioralendpointsandoutcomes.Amultimodalapproach istheproposedbestpracticebecausetheseoutcomes’totalityrevealsanoverallassessmentofpain.Pigletscastratedwithoutanestheticsoranalgesicsdemonstrateseveralbehavioralchangesindicativeofpain,asdemonstratedby increasedpain-specificbehaviorspost-procedureanddeviations

chunk 16 · 444 tokens

anestheticsoranalgesicsdemonstrateseveralbehavioralchangesindicativeofpain,asdemonstratedby increasedpain-specificbehaviorspost-procedureanddeviations inmaintenancebehaviorandpigletactivity.Pigletscastratedwithoutpaincontroldemonstrateincreasedtrembling,rumpscratching,prostration,andtailjamming/wagging(Hayetal.,2003),allofwhichindicatepain.Also,nurs-ing(McGloneetal.,1993)andsocialbehaviorsdecreasepost-procedure,andgreaterdurationandintensityoffightingwith penmatescanalsobeobserved(Hayetal.,2003;LlamasMoyaetal.,2008;Leidigetal.,2009;Sutherlandetal.,2010;Sutherlandetal.,2012).Lastly,areductioninactivitylevels(i.e.increasedinactivity)isacommonlynotedbehavioralresponse postcastration(McGloneetal.,1993;Hayetal.,2003;LlamasMoyaetal.,2008).Amongstudiesthathaverecordedpigletactivityandpainbehaviorfollowingcastration,currentbehaviorsamplingmethod- ologieshavenotbeenvalidated,andthereisconsiderablevari- ationinthemethodologiesused(O’Connoretal.,2014).Forexample,whenassessingactivityandmaintenancebehaviorsincastratedpiglets,McGloneetal.(1993)usedcontinuousliveobservationsfor6hfollowingcastrationandfoundreducedsuck- lingandstandingandincreasedlyingtimeincastratedpiglets comparedtonon-castratedlittermates.Incomparison,Hay etal.(2003)alsousedliveobservationbututilizedscansamplingat10-minuteintervalsandfoundthatcastratedpigletshad reducedactivityattheudderandspentmoretimeinactive(i.e. lying,sitting,orstanding).Similarly,LlamasMoyaetal.(2008)recordedpigletbehaviorfor3hintheafternoononthedayof castrationusing3-minutescansamplesandfoundthatcastrated pigsspentlesstimewalkingthansham-handledpiglets.These fewexampleshighlightthewidevariationinbehavioralmethod- ologiesusedtoassesscastrationpain.Bethatasitmay,allthese studieswereabletoquantifydeviationsinpigletbehavior

chunk 17 · 418 tokens

hlightthewidevariationinbehavioralmethod- ologiesusedtoassesscastrationpain.Bethatasitmay,allthese studieswereabletoquantifydeviationsinpigletbehavior post-procedure.Toaddresslimitationsassociatedwithbehavioralmethodolo-giesusedincastrationstudies,validatingtheaccuracyofdifferent behavioralsamplingmethodologiesisneeded.Also,developinga pigletbehavioralpainscaleusingbehaviorsthatcanbereliably measuredbymultiplepeopleandaresensitivetodetectingpain wouldbevaluable.Giventhelackofvalidatedbehaviormethod- ologiesforpigs,theauthorsrecommendthefollowingmethod- ologyforallbehavioralobservations,regardlessofhowbehavior dataarecollected(i.e.manualorautomatedmethods).Piglets shouldbeindividuallyidentifiedandfilmedintheirhomepens Table1.(Continued.)ReferencePigletageAnalgesiaLocalanesthesiaMeasuredoutcomeDrugefficacyaViscardiandTurner(2018a)5d0.04mgkg−1buprenorphineIM–PainbehaviorYesActivitylevelYesFacialgrimacingYes VocalizationNoViscardiandTurner(2018b)5d0.4mgkg−1meloxicamIMor1.0mgkg−1meloxicamIMor6.0mgkg−1ketoprofenIM–PainbehaviorNoActivitylevelNoFacialgrimacingNoVocalizationNoBurkemperetal.(2019)3–5d1.0mgkg−1meloxicamoral–PainbehaviorNoYunetal.(2019)5d0.4mgkg−1meloxicamIMorisoflurane(1.5%)withmeloxicamIM0.5ml2%lidocaineintratesticularPainbehaviorNoIV=intravenous;EEG=electroencephalogram;IM=intramuscular;M=meloxicam;L=lidocaine;CRP=C-reactiveprotein.aDrugefficacyisdefinedassuccessful(yes)iftreatmentadministrationminimizedoutcomemeasuresorsignificantlyminimizeddeviationstobehavioralandphysiologicalindicatorsofpain(Dzikamunhengaetal.,2014).AnimalHealthResearchReviews167 https://doi.org/10.1017/S1466252321000141 Published online by Cambridge University Press

chunk 18 · 441 tokens

continuallyovera24-hourperiodforthreedays.Identificationoftheindividualpigletswouldbeviaauniqueidentifierconsisting ofanumberandaletter,withtheletterrandomlyassignedand representsthepiglet’streatment.Thenumberidentifierisplacedonbothbacklegsandtheletteronthepiglet’sforeheadandbackusingablackpermanentmarker.Themarkingsmustberefreshed twicedailyduringstudyprocedures.Thevideoshouldbecap- tureddigitally,andbehavioralsoftwareprogrammingutilizedto capturedata.Automatedtechnologiesformeasuringpigletactivityatcastra-tionhavenotbeenusedtodatebuthavethepotentialtoprovide morereliabledataandasignificantreductioninlabor (Nasirahmadietal.,2017).Forexample,accelerometers(ped-ometers),radio-frequencyidentification,andvisualtrackingsys- temshavebeenusedtoassessactivityinolderpigsandother livestockspecies(Currahetal.,2009;Kashihaetal.,2013;Kulikovetal.,2014).Comparedtomosttrackingdevices,thesmallpigletsizepresentsachallenge,butproblemsassociated withthesetechnologiesaredecreasingwithcontinuingimprove- mentsintheirsize,accuracy,andaffordability.Therefore,while automatedsystemsmayhelpidentifyeffectivepaincontrolmea- sures,theyarenotcurrentlyrecommendedforefficacyevaluation ofpainmitigationstrategiesatcastrationastheyrequirefurther validationforuseinpiglets.PigletbehaviorandactivityobservationswillbecollectedusingcontinuousbehaviorsamplingonDay0(daybeforecastration,or baseline),Day1(0–24hpost-treatmentadministration),andDay2(24–48hpost-treatmentadministration).Behaviorstobemea-suredaredefinedintheethograminTable2.BasedonpreviousworkoutlinedinTable1,theauthorsrecommendthesamplesizeforbehavioralobservationsis10–20piglets/treatment.Beforetrialinitiation,observersmustbetrainedtoensurebehavioraldataare recordedconsistently.Toensureinter-observerreliability,a2-h

chunk 19 · 447 tokens

–20piglets/treatment.Beforetrialinitiation,observersmustbetrainedtoensurebehavioraldataare recordedconsistently.Toensureinter-observerreliability,a2-h subsetofcontinuousvideoisselectedatrandom,observed,and compareduntil90%accuracyisachieved(Rossetal.,2019).TheethogramdefinedinTable2shouldbeusedforallobser-vationaldatacollected.Allbehaviors(Table2)shouldbecollectedcontinuouslybytwotrainedobserversutilizingsoftware(e.g. ObserverXTprogram)oranydatacollection,coding,andana- lysistoolthatiseasilyvalidatedwithatimeanddatestamp. Videosarerandomizedandassignedtoobserverswhoareblind totreatmentandtimepoints.Inter-observerreliabilityisassessed beforedatacollectionandatthree-timepointsduringthebehav- iorscoringperiod.Throughoutthetrial,allinter-observerreli- abilitytestsshouldproduceanR-valueabove0.9,indicating excellentagreementbetweenscorersandnosignificantdrift throughoutthescoringperiod(Parketal.,2020).Analysisshouldbecompletedinarepeatedmeasureslinearmixedmodelwiththe Poissondistribution.InfraredthermographyIRTisanon-invasivemethodofdetectingtheamountofinfrared energy(heat)anobjectradiatesandcanbeusedtomeasureskin temperaturechangesassociatedwithactivationofthesympathetic nervoussystem.Whenananimalisstressedorinpain,thesym- patheticnervoussystembecomesactivated,whichcausesvaso- constrictionandashiftinbloodflowfromtheskintothe organs.Thebloodflowchangeresultsinalossofheatinthe body’speripheryanddecreasedskintemperature(Stewartetal.,2005;Batesetal.,2014).Stewartetal.(2010)foundthateyetemperaturemeasuredusingIRTsignificantlyincreasedinresponsetocastration.Thisresponsewasreducedwhencalvesweregivenaninjectionoflocalanestheticintothetestesandscrotum7minutesbeforecas- tration.Also,SHAMproceduresonlycausedaminorchangein eyetemperaturecomparedtothepainfulprocedures(Stewart

chunk 20 · 448 tokens

eticintothetestesandscrotum7minutesbeforecas- tration.Also,SHAMproceduresonlycausedaminorchangein eyetemperaturecomparedtothepainfulprocedures(Stewart etal.,2010).TheseresultssupportthatIRTcanbeusedasanobjectivemeasureoftheanimal’sresponsetoapainfulstimulus.Inpigs,IRThasbeenusedtodetectdisease(Cooketal.,2015),stress(Schaeferetal.,2002;Magnanietal.,2011;Sutherlandetal.,2015),andpain(Batesetal.,2014).Specifictopain,Batesetal.(2014)foundthatpigsgivenanalgesiabeforecastra-tionhadgreatercranialskintemperatures,asmeasuredusing IRT,thancastratedpigsthatdidnotreceivepainrelief.AchangeinbloodflowduringstresscanbedetectedusingIRTonspecificbodyregionssuchastheeyeincattle(Stewart etal.,2010)andhorses(Bartoloméetal.,2013),thecombinpoultry(Moeetal.,2012),andhead,snout,vulva,andteatsinpigs(Batesetal.,2014;Sutherlandetal.,2015).Batesetal.(2014)measuredchangesinskinsurfacetemperatureusingIRTonthetopofthecranium,ears,andsnoutinpigscastrated withandwithoutanalgesiaandfoundthatcranialskintempera- turewasthemostreliableanatomicallocationforassessingpain inpigletsinresponsetocastrationduetosignificanttemperature variabilityinotherlocations.Furthermore,Sutherlandetal.(2015)investigatedthepotentialforIRTasanon-invasivemeas-ureofstressinpigsandcomparedwhethertheeyeorthesnout wasamoresensitiveregiontomeasurestress.Temperature changessuggestedthattheeyemaybeamorereliableareatoassessstressthanthesnoutinpigs.TheliteratureshowsthatIRTcanbeusedtomeasureanimalpain;however,furthervalid- ationofthistechnique’smethodologyisneeded,includingthemostreliableanatomicalsiteforassessingpain.Basedondatafrompreviousstudies(Stewartetal.,2010;Batesetal.,2014),assuminga5%significancelevel,and80%power,theauthorsrecommendasamplesizeof20piglets/treatmentbased onanexpecteddifferenceincranialskintemperatureofone

chunk 21 · 393 tokens

,assuminga5%significancelevel,and80%power,theauthorsrecommendasamplesizeof20piglets/treatmentbased onanexpecteddifferenceincranialskintemperatureofone degreeCelsiusandastandarderrorofthemeanof0.1degrees Celsiusat12haftercastration,assumingdataareanalyzed using2-samplet-tests.CortisolCortisolisabiomarkercommonlyusedtomeasurestressand paininanimals.Severalpainfulhusbandryprocedures(e.g.cas- tration,taildocking,anddehorning)havebeenshowntocause anincreaseincortisolconcentrationsinseveralspecies(e.g. sheepandcattle)(Dinnissetal.,1997;Kentetal.,1998;McMeekanetal.,1998;Sutherlandetal.,1999,2002;Staffordetal.,2002)includingsurgicalcastrationandtaildockinginpigs(Prunieretal.,2005;Carrolletal.,2006;Sutherlandetal.,2012).Numerousstudieshaveshownthatsurgicalcastrationcausesasignificantandmarkedincreaseinpigs’cortisolconcen-trations(Prunieretal.,2005;Carrolletal.,2006;Sutherlandetal.,2010,2012,2017).However,handlingaloneonlycausesaslightbutnon-significantincreaseincortisol(Prunieretal.,2005),sug-gestingthattheincreaseincortisolinresponsetocastrationis predominantlyduetothepainoftheprocedureandnotthestress ofhandling.Moreover,inpigsgivenanalgesia(e.g.lidocaineora non-steroidalanti-inflammatorydrug),thecortisolresponseto surgicalcastrationwasreduced(Kluivers-Poodtetal.,2012;Batesetal.,2014).Cortisoliscommonlymeasuredinplasma,serum,orsaliva.Aftersurgicalcastrationinpigs,plasmacortisolconcentrations peakbetween15and60minandreturntobaselinelevelsbetween168AngelaBaysingeretal. https://doi.org/10.1017/S1466252321000141 Published online by Cambridge University Press

chunk 22 · 369 tokens

120-and180-minutespost-procedure(Prunieretal.,2005;Sutherlandetal.,2012,2017).Therefore,toevaluatetheefficacyofdifferentpainmitigationstrategiestoreducethepaincaused bysurgicalcastration,cortisolconcentrationsshouldbemeasured immediatelybeforetheproceduretoassessbaselinelevels, between15and60minpostcastrationtoassesschangesin peakcortisollevelsandthenagainat120mintoconfirmthat levelshavereturnedtobaseline.Cortisolconcentrationsinpigs canbemeasuredusingvalidatedin-houseradioimmunoassay (RIA)orenzymeimmunoassay(EIA)techniques.However,in therecentliterature,cortisolconcentrationsinpigshavebeen morecommonlymeasuredusingcommerciallyavailableRIA kitssuchasCoat-a-Count(SiemensMedicalSolutions Diagnostics[formallyDiagnosticProductsCorp],LosAngeles,California)(Kluivers-Poodtetal.,2012;Batesetal.,2014)andEIAkitssuchasAssayDesigns(AnnArbor,Michigan)(Carroll etal.,2006;Sutherlandetal.,2010,2012)andIMMULITE/IMMULITE1000Cortisol(GlobalSiemensHealthcare, Erlanger,Germany)(Sutherlandetal.,2008).Changesincortisolshouldbeincludedasabiomarkerforpaininpigsasithasreliablybeenshowntoincreaseinresponseto painandbereducedorabolishedinresponsetodifferentpain mitigationstrategiesaftercontrollingforconfoundingfactors suchashandling,restraint,andtissuetrauma(Sheiland Polkinghorne,2020).Besides,cortisolcanbereliablymeasuredinsucklingpigs’serumorplasmausingcommerciallyavailableRIAorEIAkits.However,whencommercialdiagnostickits developedandvalidatedforthediagnosisofendocrinedisorders

chunk 23 · 400 tokens

pigs’serumorplasmausingcommerciallyavailableRIAorEIAkits.However,whencommercialdiagnostickits developedandvalidatedforthediagnosisofendocrinedisorders Table2.BehavioralethogramforpigletsadaptedfromHayetal.(2003)CategoryBehaviorDefinitionNon-specificbehaviorsUddermassageorsucklingNoseincontactwiththeudderorteatinthemouth.Vigorousandrhythmicupanddownheadmovementsorsucklingmovements.LookingforteatAttemptstofindateatbywalkingandpushingotherpigletswhilemostoftheothersaresuckling. NosingThesnoutisclosetoorincontactwithasubstrateorapen-mate.Snoutmovementsmaybeobserved. Belly-nosingRepeatedupanddownmassagemovementswiththesnoutontoanotherpigletorthesow(excepttheudder).ChewingorlickingRubbingthetongueoverornibblingatlittermates(ears,tailorfoot,etc.),substrates,floor,orpenwalls. PlayingHeadshaking,springing(suddenjumpingorleaping),runningwithverticalandhorizontalbouncymovements.Itcaninvolvepartners(gentlenudgingorpushing,mounting,chasing).AggressionForcefulfighting,pushingwiththehead,orviolentlybitinglittermates. WalkingSlowlymovingforwardwithonelegatatime. RunningTrotorgallopwithoutasuddenchangeindirectionorspeed.AwakeinactiveNospecialactivitybutawake.Lying,sitting,orstanding.SleepingLyingdown,eyesclosed.PainbehaviorProstrateAwake,sittingorstandingmotionless,withtheheaddown,lowerthanshoulderlevel.HuddledupLyingwithatleastthreelegstuckedunderthebody. StiffnessLyingwithextendedandtensedlegs.TremblingShiveringaswithcold.Theanimalmaybelying,sitting,orstanding.SpasmsQuickandinvoluntarycontractionsofthemusclesundertheskin ScratchingScratchingtherumpbyrubbingitagainstthefloororthepenwalls.

chunk 24 · 287 tokens

g,sitting,orstanding.SpasmsQuickandinvoluntarycontractionsofthemusclesundertheskin ScratchingScratchingtherumpbyrubbingitagainstthefloororthepenwalls. TailwaggingTail’smovementsfromsidetosideorupanddown.PosturesLaterallyingMotionless;bodyweightsupportedbyside.Shoulderincontactwiththefloor.VentrallyingMotionless;bodyweightsupportedbybelly.Sternumincontactwiththefloor.SittingMotionless;bodyweightsupportedbyhind-quartersandfrontlegs.StandingMotionless;bodyweightsupportedbythefourlegs. KneelingMotionless;bodyweightsupportedbyfrontcarpaljointsandhindlegs.LocationUdderCloseto(<10cm)orincontactwiththeudder.Sow’sbackCloseto(<10cm)orincontactwithsow’sback.HeatmatOntheheatmat.SocialcohesionIsolatedAsidefromotherpiglets,aloneorwithonepen-mateatthemost.Adistanceofatleast40cm(aboutthewidthoftwopiglets)separatestheanimalfromtheclosestgroupoflittermates.DesynchronizedActivitydifferentfromthatofmost(atleast75%)littermates(e.g.sleepswhilemostotherlittermatessuckle).OtherOtherThepig’sbehaviorcannotbedetermined,ortheanimalwasnotseen.AnimalHealthResearchReviews169 https://doi.org/10.1017/S1466252321000141 Published online by Cambridge University Press

chunk 25 · 379 tokens

suchashyperadrenocorticismarerepurposedasanalyticalmeth-odstomeasuretheconcentrationsofbiomarkersinhealthyani- malsundergoingpainfulprocedures,researchersmustensure thatthediagnostickitvalidationdataprovidedbythemanufac- turerisreliableandaccurate.Furthermore,itisagoodscientific practicetovalidatethediagnostickit’sperformanceinthefacilityconductingthesampleanalysis.Usingdatacollectedfrompreviousstudies(Sutherlandetal.,2010,2012,2017;Bonastreetal.,2016),assuminga5%signifi-canceleveland80%power,theauthorsrecommendusingasam- plesizeof20piglets/treatmentbasedonanexpecteddifferencein serumcortisolconcentrationsof20ngmL−1andastandarderrorofthemeanof5ngmL−1at60minaftersurgicalcastration,assumingdataareanalyzedusing2-samplet-tests.AncillaryoutcomesStridelength,contactpressure,contactarea,andstancephaseduration Objectivegaitparameters,measuredusingacommerciallyavail- ablefloormat-basedpressure/forcemeasurementsystem (MatScan,Tekscan,Inc,SouthBoston,Massachusetts),were usedinconjunctionwithlamenessscoresasprimaryendpoints inapivotalstudythatsupportedFDAapprovalofthefirstanal- gesicdruglabeledforuseincattleintheUnitedStates(Banamine Transdermal,MerckInc,Madison,NewJersey)(USFoodand DrugAdministration,2017).Pressurematanalysisisrecognizedwithinavalidatedfootrotmodelasareliablepainassessment endpointincattle,thussatisfyingtheFDAGuidanceDocument 123requirements(USFoodandDrugAdministration,2006).Pressuremattechnologyhasbeenusedtorecordandanalyzenat- urallyoccurringorexperimentallyinducedchangesingaitincat-

chunk 26 · 425 tokens

nts(USFoodandDrugAdministration,2006).Pressuremattechnologyhasbeenusedtorecordandanalyzenat- urallyoccurringorexperimentallyinducedchangesingaitincat- tleandswineduetolameness(Kotschwaretal.,2009;Schulzetal.,2011;Karrikeretal.,2013;Coetzeeetal.,2014;Pairis-Garciaetal.,2015)andsurgicalcastration(Nasirahmadietal.,2017;Kleinhenzetal.,2018).Takentogether,thesedatasupporttheassessmentofstridelength,contactpressure,contactarea,andstancephasedurationusingthepressurematinthepro- posedefficacystudy.Oneoftheproposedstudy’skeyoutcomeswouldbetocomparethepressurematoutcomeswithbehavioral andphysiologicaloutcomes.Thepressurematwillbecalibrateddailyusingtheexpectedbodyweightofthepiglets,andeachtimethecomputersoftware isengagedusingaknownmasstoensuretheaccuracyofthemea- surementsateachtimepoint.Thepressuremat,measuring6–8feetinlength,mustbesetuponaflatsurfacewherepigletscan bedirectedtowalkatasteadypaceacrossthematsothatthedis- tancebetweenmultiplefootfalls,pressure,andstancecanbemea- sured.Footfallsarerecordedwhenthefootstrikestheloadedor ‘contact’sensingelementsinsideameasurementbox.Researchgradesoftware(HUGEMATResearch5.83,Tekscan,Inc.,South Boston,Massachusetts)isusedtodeterminethecontactpressure, contactarea,stancephaseduration,andstridelength.Thewalk- ingpig’svideoiscaptureddigitallyandsynchronizedtoensureconsistentgaitbetweenandwithinpigletsforeachtimepoint. Readingsaretakenbeforecastrationandat6,12,24,and48h aftercastration.Thepercentchangefrombaselineforallmea- sureswillbecalculatedandanalyzedstatisticallyusinga mixed-effectsmodel.Beforetrialinitiation,observersmustbe trainedtoensurestridelengthdataisrecordedconsistently.The observerswillachieve80%inter-observerreliabilityandbe

chunk 27 · 445 tokens

.Beforetrialinitiation,observersmustbe trainedtoensurestridelengthdataisrecordedconsistently.The observerswillachieve80%inter-observerreliabilityandbe blindedtotreatmenttocontrolforobserverbias.Theanalysis’soutputcanbeconvertedtoPDF,allowingtheoutcomestobereconstructedafterthestudyiscompleted.Usingdatacollectedfromanunpublishedpilotstudy,assum-inga5%significanceleveland80%power,theauthorsrecom- mendusingasamplesizeof30piglets/treatmentbasedonan expectedpercentchangeinfrontstridelengthfrombaselinemea- surementsof30%,astandarderrorofthemeanof10%assuming thedataareanalyzedbyANOVA.BloodbiomarkersSeveralcirculatingbiomarkerstargetingtheindirectassessmentofpainandstresshavebeenmeasuredinpigletsatthetimeofcas- tration.Specifically,theseincludemarkersoftheneuroendocrine response(e.g.corticotropin,β-endorphins,epinephrine,norepin-ephrine,andsubstanceP),inflammatoryresponse(e.g.haptoglo- bin,c-reactiveprotein,serumamyloidA,andprostaglandinE2), andadrenocorticalresponse(e.g.cortisol)(Wearyetal.,2006;Dzikamunhengaetal.,2014).Also,immuneresponseassessmentshavebeenmadeusinghematologicalendpointssuchasneutrophil tolymphocyteratio(N:L)derivedfromacompletebloodcount. Thesebiomarkershavebeencorrelatedwithstressbecausecortisol releaseistypicallyrapidanddifficulttoquantify,buttheasso- ciatedstressleukogramlastslongerandcouldbelesstime- sensitive.Also,theproductionofunconjugatedpterins(neopterin andbiopterin)havebeenassociatedwithstressfulsituationssuch aspigletcastration(Marsáleketal.,2011;Maršáleketal.,2015).Bloodbiomarkersareindirectmeasuresofpainandinflamma-tion.Furthermore,manyoftheseoutcomeshavelowspecificity andcanbealteredbyotherfactorssuchashandlingstress. Theseoutcomesaresusceptibletoconfoundingbyotheraspects oftheexperiment,specificallybloodsamplingandhandling.

chunk 28 · 446 tokens

lteredbyotherfactorssuchashandlingstress. Theseoutcomesaresusceptibletoconfoundingbyotheraspects oftheexperiment,specificallybloodsamplingandhandling. Mostalltheanalyticalmethodshavenotbeenvalidatedto GoodLaboratoryPracticeorGoodClinicalPracticespecifications. Attemptsatvalidatingtheseoutcomesinpigletshavebeencon- foundedbychallengeswithextractingtheanalytesfromplasmaandserum.Additionalchallengessurroundthesamplecollec-tion’soptimaltimingtoensurethattheoutcomesarecorrelatedwiththepainfulevent.Aninitialscreeningofbloodbiomarkers relativetothepainfuleventmayhelpidentifyoutcomesthatwar- rantfurtherinvestigationandvalidation.Bloodbiomarkerassessmentiscurrentlypredicatedusingimmunoassaysorautomatedhematologicalmethodsvalidated forhumanmedicine.Thecollectionprotocol(timingandamount ofbloodneeded)willdependonthestudy’sbiomarkers.Aftercollection,theanalytestabilityremainsasignificantchallenge;therefore,samplesmustbestoredoniceand,insomecases,liquid nitrogen.InthecaseofneuropeptidessuchassubstanceP,add- itionalstepsmustbetakentoensurethatserineproteasesdo notdegradeaftercollection(Mosheretal.,2014).Onemethodinvolvestheadditionofbenzamidinehydrochloride,aprotease inhibitor,at1mMmL−1ofwholebloodcollectedinEDTA.Hematologicaloutcomesarealsomeasuredusingbloodcollected inEDTA,whileacutephaseproteins,suchashaptoglobin,are collectedinserumtubes.Previousstudiesinvestigatingbloodbio- markershavegenerallyenrolledbetween10and20piglets/treat- ment(Raultetal.,2011;Sutherlandetal.,2012;Dzikamunhengaetal.,2014).Initially,allofthebiomarkersmen-tionedabovewouldbemeasuredtodetermineusefulnessasan indirectoutcomevariableinthemultimodalprotocoltomeasure analgesicefficacy.Uponidentificationofthebiomarker(s)that consistentlycorrelateswiththeprimaryoutcomemeasuresof170AngelaBaysingeretal.

chunk 29 · 60 tokens

otocoltomeasure analgesicefficacy.Uponidentificationofthebiomarker(s)that consistentlycorrelateswiththeprimaryoutcomemeasuresof170AngelaBaysingeretal. https://doi.org/10.1017/S1466252321000141 Published online by Cambridge University Press

chunk 30 · 172 tokens

pain,oneormorebiomarkerswouldbeutilizedinthemulti-modalprotocol.PigletgrimacescaleFacialgrimacescalesareanovel,non-invasivetoolforpain assessmentusingquantifiablechangestofacialfeaturestodetectpain.Theyhavebeendevelopedfornon-verbalhumansaswellasmanyanimals,includingmice,rats,rabbits,horses,sheep, lambs,cattle,andpiglets(Langfordetal.,2010;Herretal.,2011;Sotocinaletal.,2011;Keatingetal.,2012;Costaetal.,2014;Gleerupetal.,2015;DiGiminianietal.,2016;Guesgenetal.,2016;McLennanetal.,2016;Hägeretal.,2017).APiglet Fig.1.PigletgrimacescaledevelopedbyViscardietal.(2017).AnimalHealthResearchReviews171 https://doi.org/10.1017/S1466252321000141 Published online by Cambridge University Press

chunk 31 · 429 tokens

GrimaceScale(PGS)developedbyViscardietal.(2017)describedchangestothreefacialactionunitsinresponsetopiglet surgicalcastrationandtaildockingpain(Fig.1).Afacialgrimaceinpigletsischaracterizedbynarrowingtheorbitalarea(eyes squeezingshut),earspulledbackagainstthehead,andapromin- entbumporbulgeonthesnoutresultingfromcheektightening. AccordingtothePGS,themaximumgrimacescoreis5,andit hascorrespondedwelltodisplayedpainbehaviors(e.g.an increaseinpainbehaviorcorrespondedtohigherfacialgrimacing inpiglets)(ViscardiandTurner,2018a,b).Althoughanincreaseinpigletfacialgrimacinghasbeencorre-latedtoadecreaseinactivitylevelandcorrespondstoanincrease inpainbehavior(Viscardietal.,2017;ViscardiandTurner,2018a,b),thePGShasnotbeenvalidatedasapainassessmenttool.Thisproposedstudymethodologywilldetermineiffacial grimacingcanbecorrelatedtopainbehaviororothernon- invasiveoutcomemeasures(e.g.IRT).ThePGShasonlybeen usedtoretrospectivelyassesspainandanalgesiaefficacybyscor- ingstill-imagesofpigletfacialexpressionsextractedfromvideo recordings.Individualsusedtoscorefacialexpressionswere undergraduate,graduate,orveterinarystudents.Toimproveits practicalityforon-farmuse,thePGSshouldbevalidatedforreal- timedetectionofpainandproducersandswineveterinarians’abilitytousethePGStoidentifypaininpigletsaccurately.Previousworkusinggrimacescaleshavefound1–3facialimagescapturedperanimalpertimepointwassufficienttoassess painandanalgesicefficacy(Sotocinaletal.,2011;Costaetal.,2014;Milleretal.,2015;MillerandLeach,2015a,b;Milleretal.,2016;ViscardiandTurner,2018a).Thesestudiesalsoused2–5individualstoscorefacialexpressions.Researchersaver-agedtheindividuals’resultingscoresbeforeanalysisandcon-ductedinter-observerreliabilityteststoensurescoring

chunk 32 · 438 tokens

alstoscorefacialexpressions.Researchersaver-agedtheindividuals’resultingscoresbeforeanalysisandcon-ductedinter-observerreliabilityteststoensurescoring consistency.BodyweightChangesinbodyweightcanbeusedasanindirectmeasureofthepainexperiencedbypigletsatcastration.Multiplestudiesinarangeofspeciesdemonstratethatwhenanimalsexperience pain,feedconsumptiondecreases,resultinginareductionin bodyweightandaveragedailygain(ADG)(Malavasietal.,2006).Althoughthereislittleevidenceofthelong-termimpactofcastrationonbodyweightgaininpiglets,reductionsinADG duetocastrationhavebeenfoundinthedaysfollowingthepro- cedure.Kiellyetal.(1999)foundthatpigscastratedat3daysofagegainedlessweightthanweight-matchedcontrolsoverthe 3daysfollowingcastration,whilethosecastratedat14daysof ageshowednodifferenceinADGcomparedtocontrols,suggest- ingthatdelayedcastrationmaybenefitpiglets.Incontrast,Hay etal.(2003)comparedthebodyweightsofpigletscastratedat5daysofageandshamhandledcontrolstwiceperdayfor4 daysaftertreatmentandfoundnodifferencesinADG.Moststudieshavemeasuredbodyweightsatweaningandfoundnoeffectofcastrationwithorwithoutpaincontrolon bodyweight(Kiellyetal.,1999;Cassaretal.,2014;Burkemperetal.,2019).Contrarytothis,McGloneetal.(1993)foundthatpigletscastratedatonedayofagehadlowerweaningweights thanthosecastratedat14days,whilefemalepigswereintermedi- ate.Becausefemalepigswereusedascontrolsratherthansham- handledmales,thisresultisdifficulttointerpret.Amorerecent studybyMoralesetal.(2017)comparedADGinover3000castratedversusintactmalesandcategorizedpigletsbasedonini-tialbodyweightaslow(lowest25%),medium,orhigh(highest25%).Thestudyfoundthatamongheavierpigs,castrateshadlowerADGatweaningcomparedtonon-castratedpigs. Furthermore,low-andmedium-weightpigletsthatwerecastrated

chunk 33 · 445 tokens

).Thestudyfoundthatamongheavierpigs,castrateshadlowerADGatweaningcomparedtonon-castratedpigs. Furthermore,low-andmedium-weightpigletsthatwerecastrated hadahigherlikelihoodofpre-weaningmortalitythantheirnon- castratedlittermates(Moralesetal.,2017).Whilebodyweightchangesarenotadirectmeasureofpain,initialbodyweightsshouldbeconsideredforenrollmentpur- poses.Controlanimalsshouldbenon-castratedmalelittermates ratherthanfemales.Pigletsweighing1.5kgorlessshouldbe excluded.MortalityrateThemortalityrateisnotanindicatorofpain,andintervention efficacytrialsarenottypicallydesignedtodetectstatisticaldiffer- encesbetweentreatments.However,whensupplementedwithacompletenecropsyofthepigsthatdie,themortalityratecanbeusedasanon-specificindicatorofnegativeimpactsonhealth, toxicitywhenchemicalinterventionsareapplied,andsecondary complicationsthatmightinfluencetheadoptionofaninterven- tion.Assomeunrelatedbaseline,mortalityislikelytooccurin mostpopulations,andoccasionaleuthanasiaofanimalsiswar- rantedforunrelatedreasons.Mortalityisanon-specificendpoint, andastepwiseevaluationprocessshouldbeimplementedtouse studyresourceseffectively.First,potentialphysiologicalimpactsthatcanprogresstodeathandarespecifictotheinterventionbeingcomparedshouldbe identifiedaprioriatthestartofthestudy.Anyspecificnecropsylesionsandpost-mortemdiagnostictestingtoconfirmorrefute theintervention’sinvolvementinthemortalitymustberecorded.Forexample,ifadruginterventionhasthepotentialforharmful toxicity,theappropriatetissue,diagnostictest,andthetesting laboratoryshouldbeidentifiedandincludedinthestudyprotocol asastandardcomponentofthenecropsyevaluation.Thesecondstepistorecordthepig’sidentity,timeanddateobserved,andwhetherthedeathwastheoutcomeofeuthanasia oroccurrednaturally.Incaseswhereeuthanasiawasthecause

chunk 34 · 364 tokens

ndstepistorecordthepig’sidentity,timeanddateobserved,andwhetherthedeathwastheoutcomeofeuthanasia oroccurrednaturally.Incaseswhereeuthanasiawasthecause ofdeath,thereasonforeuthanasiashouldberecorded.Thethirdstepistoperformacompleteexternalexamandnec-ropsyprocedure,asoutlinedbyTorrison(2012).Theobservationofgrosslesionsortheconfirmationofabsenceshouldberecorded forallthemainbodysystems.Theuseofachecklist(Fig.2)bytrainedobserversisnecessarytomaintainconsistencyofevalua- tionsanddatacollection.Supposegrosslesionsindicateaspecificorgansystem’sinvolvement,butacauseofdeathcannotbedeterminedbythe necropsyprocedurealone.Inthatcase,thefourthstepisto employtheappropriatetissuecollectionasoutlinedbyTorrison (2012),followedbysubmissiontoaqualifieddiagnosticlabora-torywithappropriatehistoryandcontext.Allcorrespondencewiththediagnosticlaboratoryandtestresultsshouldbecopied intothestudyrecord.DiscussionItiswelldocumentedthatpigletsexperiencepainassociatedwithcastration,andeffortstomitigatethispainshouldbeexplored. Previousstudiesonpainmitigationatcastrationhavetypically includedsimilartechniquesandoutcomes,butwidevariation inexperimentaldesignanddatacollectionapproachesgivenon- comparableresultsandhindersacomprehensiveinterpretation ofthescience(SheilandPolkinghorne,2020).Thispaperdescribesamethodologytoassessbehavioral,physiological,and172AngelaBaysingeretal. https://doi.org/10.1017/S1466252321000141 Published online by Cambridge University Press

chunk 35 · 392 tokens

neuroendocrinechangesassociatedwithpaininpigletsresultingfromsurgicalcastration.Themethodologyisbeingfurtherdevel- opedintoaresearchprotocoltemplatetofacilitateandguidethe validityandreliabilityofendpointstoevaluatedrugs’effectivenesstocontrolpost-operativepainin3-to5-day-oldpigletsfollowing surgicalcastration.Fortheexperimentaldesignandthepurposesofendpointval-idation,theauthorsrecommendfourprimarytreatments,includ- ing(1)shamcastration,(2)shamcastrationwiththe‘goldstandard’intervention,(3)surgicalcastrationwithaplacebocon-trol,and(4)surgicalcastrationwiththe‘goldstandard’interven-tion.Piglet(withinlitter)istheexperimentalunitwithall treatmentsrepresentedwithineachlitteratleastonce. Outcomesbeingdirectlyrelatedtoclinicalsignsofpain(observationofpainbehaviors,activitytracking,IRT,andcortisolconcentrations)andancillaryoutcomes(stridelength,bloodbio- markers[includingneuroendocrine,inflammatory,immuno- logical,andstressresponsemarkers],pigletgrimacescale,body weight,andmortalityrate)beingeitherdirectlyrelatedtoclinical signsofpainorindirectlyrelatedtoclinicalsignsofpainand lendingsupporttothedirectoutcomeswillbemeasured,vali- datedandanalyzed.ConclusionTheexperimentaldesignandmeasurableoutcomesselectedfromthevalidationstudyareintendedtopromoteaconsistent approachtodeterminingmoreeffectivetherapiesforpain Fig.2.Checklistofrelevantmortalityinformation,necropsyobservations,includingorgansystemstobeobservedandlesionstobeconsidered.AnimalHealthResearchReviews173 https://doi.org/10.1017/S1466252321000141 Published online by Cambridge University Press

chunk 36 · 236 tokens

mitigation.Thispapersupportstheinclusionofspecificoutcomesinthevalidationstudyandsummarizestheneedforfurtherval- idationofemergingoutcomes.Thedevelopmentofsimilarproto- colsfordeterminingthevalidityandreliabilityofendpointsto evaluatetheefficacyofpainmitigationtherapiestargetedto otherpainfulproceduresorconditionsinswine,suchastaildock- ingorlameness,shouldbeconsidered.Authorcontributions.Conceptualization,SRWandAD;methodology,AKB,SRW,JB,JFC,SC,AD,LAK,M.P-G.,MAS,AVV;writing-originaldraftpreparation,AKBandSRW;writing-reviewandediting,AKBSRWandM.P-G.Allauthorshavereadandagreedtothepublishedversionofthemanuscript.Financialsupport.Nomonetaryfundingwassourcedforthedevelopmentofthismanuscript.Conflictofinterest.Theauthorsdeclarenoconflictofinterest.ReferencesBartoloméE,SánchezMJ,MolinaA,SchaeferAL,CervantesIandValeraM(2013)Usingeyetemperatureandheartrateforstressassessmentinyounghorsescompetinginjumpingcompetitionsanditspossibleinfluence

chunk 37 · 518 tokens

aeferAL,CervantesIandValeraM(2013)Usingeyetemperatureandheartrateforstressassessmentinyounghorsescompetinginjumpingcompetitionsanditspossibleinfluence onsportperformance.Animal:AnInternationalJournalofAnimalBioscience7,2044–2053.BatesJL,KarrikerLA,StockML,PertzbornKM,BaldwinLG,WulfLW,LeeCJ,WangCandCoetzeeJF(2014)Impactoftransmammary-deliveredmeloxicamonbiomarkersofpainanddistressinpigletsaftercastrationandtaildocking.PLoSOne9,e113678.BatesonP(1991)Assessmentofpaininanimals.AnimalBehaviour42,827–839.BlassEM(1994)Behavioralandphysiologicalconsequencesofsucklinginratandhumannewborns.ActaPaediatrica(Oslo,Norway)397,71–76.BonastreC,MitjanaO,TejedorM,CalaviaM,YusteA,ÚbedaJandFalcetoM(2016)Acutephysiologicalresponsestocastration-relatedpaininpiglets:theeffectoftwolocalanestheticswithorwithoutmeloxicam.Animal:AnInternationalJournalofAnimalBioscience10,1474–1481.BurkemperM,Pairis-GarciaMD,MoraesLE,ParkRMandMoellerSJ(2019)Effectsoforalmeloxicamandtopicallidocaineonbehaviorofpigletsundergoingsurgicalcastration.JournalofAppliedAnimalWelfareScience23,209–218.CarrollJA,BergEL,StrauchTA,RobertsMPandKatteshHG(2006)Hormonalprofiles,behavioralresponses,andshort-termgrowthperform-anceaftercastrationofpigsatthree,six,nine,ortwelvedaysofage.JournalofAnimalScience84,1271–1278.CassarG,AmezucaR,TenbergenRandFriendshipB(2014)Preoperativeketoprofenadministrationtopigletsundergoingcastrationdoesnotaffectsubsequentgrowthperformance.TheCanadianVeterinaryJournal55,1250–1252.ChahlLA(1996)Opioids-mechanismofaction.AustralianPrescriber19,63–65.CoetzeeJF,MosherRA,AndersonDE,RobertB,KohakeLE,GehringR,WhiteBJ,KukanichBandWangC(2014)Impactoforalmeloxicamadministeredaloneorincombinationwithgabapentinonexperimentallyinducedlamenessinbeefcalves.JournalofAnimalScience92,816–829.CookNJ,ChabotB,LuiT,BenchCJandSchaeferAL(2015)Infraredthermographydetectsfebrileandbehaviouralresponsestovaccinationofweanedpiglets.Animal:AnInternationalJournalofAnimalBioscience9,339–346.CostaED,MineroM,LebeltD,StuckeD,CanaliEandLeachMC(2014)Developmentofthehorsegrimacescale(HGS)asapainassessmenttool

chunk 38 · 270 tokens

lJournalofAnimalBioscience9,339–346.CostaED,MineroM,LebeltD,StuckeD,CanaliEandLeachMC(2014)Developmentofthehorsegrimacescale(HGS)asapainassessmenttool inhorsesundergoingroutinecastration.PLoSOne9,e92281.doi:10.1371/journal.pone.0092281CurrahJM,HendrickSHandStookeyJM(2009)Thebehavioralassessmentandalleviationofpainassociatedwithcastrationinbeefcalvestreatedwithflunixinmeglumineandcaudallidocaineepiduralanesthesiawithepineph-rine.TheCanadianVeterinaryJournal50,375–382.DiGiminianiP,BrierleyVLMH,ScolloA,GottardoF,MalcolmEM,EdwardsSAandLeachMC(2016)Theassessmentoffacialexpressionsinpigletsundergoingtaildockingandcastration:towardthedevelopment ofthepigletgrimacescale.FrontiersinVeterinaryScience3,100.DinnissAS,MellorDJ,StaffordKJ,BruceRAandWardRN(1997)Acutecortisolresponsesoflambstocastrationusingarubberringand/oracas-trationclampwithorwithoutlocalanaesthetic.NewZealandVeterinaryJournal45,114–121.DzikamunhengaRS,AnthonyR,CoetzeeJF,GouldS,JohnsonA,KarrikerL,McKeanJ,MillmanST,NiekampSRandO’ConnorAM(2014)Painmanagementintheneonatalpigletduringroutinemanagementprocedures.

chunk 39 · 553 tokens

,CoetzeeJF,GouldS,JohnsonA,KarrikerL,McKeanJ,MillmanST,NiekampSRandO’ConnorAM(2014)Painmanagementintheneonatalpigletduringroutinemanagementprocedures. Part1:asystematicreviewofrandomizedandnon-randomizedinterventionstudies.AnimalHealthResearchReviews15,14–38.EuropeanCommision(2017)Pigcastration:methodsofanaesthesiaandanalgesiaforallpigsandotheralternativesforpigsusedintraditionalpro-ducts.[Online]Availableat:https://op.europa.eu/en/publication-detail/-/publication/5fe8db00-dbb8-11e6-ad7c-01aa75ed71a1/language-en/format-PDF/source-220070556(accessed18/07/21).FestingMFW(2006)Designandstatisticalmethodsinstudiesusinganimalmodelsofdevelopment.ILARJournal47,5–14.FieldTandGoldsonE(1984)Pacifyingeffectsofnonnutritivesucklingontermandpretermneonatesduringheelstickprocedures.Pediatrics74,1012–1015.GaynorJSandMuirWW(2009)Painbehaviors.InGaynorJSandMuirWW(eds.),HandbookofVeterinaryPainManagement.2ndEdn.St.Louis,MO:MosbyElsevier,pp.62–77.GleerupK,AndersenP,MunksgaardLandForkmanB(2015)Painevalu-ationincattle.AppliedAnimalBehaviourScience171,25–32.GuesgenMJ,BeausoleilNJ,LeachM,MinotEO,StewartMandStaffordKJ(2016)Codingandquantificationofafacialexpressionforpaininlambs.BehaviouralProcesses132,49–56.HagaHAandRanheimB(2005)Castrationofpiglets:theanalgesiceffectsofintratesticularandintrafunicularlidocaineinjection.VeterinaryAnaesthesiaandAnalgesia32,1–9.HägerC,BiernotS,BuettnerM,GlageS,KeublerLM,HeldN,BleichEM,OttoK,MüllerCW,DeckerS,TalbotSRandBleichA(2017)Thesheepgrimacescaleasanindicatorofpost-operativedistressandpaininlabora-torysheep.PLoSOne12,e0175839.HanssonM,LundeheimN,NymanGandJohanssonG(2011)Effectoflocalanaesthesiaand/oranalgesiaonpainresponsesinducedbypigletcastration.ActaVeterinariaScandinavica53,34.HayM,VulinA,GéninS,SalesPandPrunierA(2003)Assessmentofpaininducedbycastrationinpiglets:behavioralandphysiologicalresponsesoverthesubsequent5days.AppliedAnimalBehaviourScience82,201–218.HermansenK,PedersenLEandOlesenHO(1986)Theanalgesiceffectofbuprenorphine,etorphineandpethidineinthepig:arandomizeddouble-blindcross-overstudy.ActaPharmacologicaetToxicologica59,27–35.HerrK,CoynePJ,McCafferyM,ManworrenRandMerkelS(2011)Painassessmentinthepatientunabletoself-report:positionstatementwithclin-

chunk 40 · 303 tokens

gicaetToxicologica59,27–35.HerrK,CoynePJ,McCafferyM,ManworrenRandMerkelS(2011)Painassessmentinthepatientunabletoself-report:positionstatementwithclin- icalpracticerecommendations.PainManagementNursing12,230–250.HornT,MarxGandvonBorellE(1999)Behaviorofpigletsduringcastrationwithandwithoutlocalanesthesia.DTW.DeutscheTierarztlicheWochenschrift106,271–274.IASPSubcommitteeonTaxonomy(1979)Painterms:alistofdefinitionsandnotesonusage.RecommendedbytheIASPsubcommitteeontax-onomy.Pain6,249.IsonSH,CluttonRE,DiGiminianiPandRutherfordKMD(2016)Areviewofpainassessmentinpigs.FrontiersinVeterinaryScience3,108.KarrikerLA,AbellC,Pairis-GarciaMD,HoltWA,SunG,CoetzeeJF,JohnsonAK,HoffSJandStalderKJ(2013)Validationofalamenessmodelinsowsusingphysiologicalandmechanicalmeasurements.JournalofAnimalScience91,130–136.KashihaM,BahrC,OttC,MoonsCPH,NiewoldTA,OdbergFOandBerckmansD(2013)Automaticidentificationofmarkedpigsinapenusingimagepatternrecognition.ComputersandElectronicsinAgriculture93,111–120.KeatingSCJ,ThomasAA,FlecknellPAandLeachMC(2012)EvaluationofEMLAcreamforpreventingpainduringtattooingofrabbits:changesin174AngelaBaysingeretal. https://doi.org/10.1017/S1466252321000141 Published online by Cambridge University Press

chunk 41 · 528 tokens

physiological,behavioralandfacialexpressionresponses.PLoSOne7,e44437.KeitaA,PagotE,PrunierAandGuidariniC(2010)Pre-emptivemeloxicamforpost-operativeanalgesiainpigletsundergoingsurgicalcastration.VeterinaryAnaesthesiaandAnalgesia37,367–374.KentJE,MolonyVandGrahamMJ(1998)Comparisonofmethodsforthereductionofacutepainproducedbyrubberringcastrationortaildockingofweek-oldlambs.VeterinaryJournal155,39–51.KiellyJ,DeweyCEandCochraneM(1999)Castrationat3daysofagetempor-arilyslowsgrowthofpigs.JournalofSwineHealthandProduction7,151–153.KleinhenzMD,VanEngenNK,SmithJS,GordenPJ,JiJ,WangC,PerkinsSCBandCoetzeeJF(2018)Theimpactoftransdermalflunixinmeglumineonbiomarkersofpainincalveswhenadministeredatthetimeofsurgicalcastrationwithoutlocalanesthesia.LivestockScience212,1–6.Kluivers-PoodtM,HouxBB,RobbenSRM,KoopG,LambooijEandHellebrekersLJ(2012)EffectsofalocalanaestheticandNSAIDincastra-tionofpiglets,ontheacutepainresponses,growthandmortality.Animal:AnInternationalJournalofAnimalBioscience6,1469–1475.Kluivers-PoodtM,ZonderlandJJ,VerbraakJ,LambooijEandHellebrekersLJ(2013)Painbehavioraftercastrationofpiglets;effectofpainreliefwithlidocaineand/ormeloxicam.Animal:AnInternationalJournalofAnimalBioscience7,1158–1162.KotschwarJL,CoetzeeJF,AndersonDE,GehringR,KukanichBandApleyMD(2009)AnalgesicefficacyofsodiumsalicylateinanamphotericinB-inducedbovinesynovitis-arthritismodel.JournalofDairyScience92,3731–3743.KulikovVA,KhotskinNV,NikitinSV,LankinVS,KulikovAVandTrapezovOV(2014)Applicationof3-Dimagingsensorfortrackingmini-pigsintheopenfieldtest.JournalofNeuroscienceMethods235,219–225.LangfordDJ,BaileyAL,ChandaML,ClarkeSE,DrummondTE,EcholsS,GlickS,IngraoJ,Klassen-RossT,LaCroix-FralishML,MatsumiyaL,SorgeRE,SotocinalSG,TabakaJM,WongD,vandenMaagdenbergAMJM,FerrariMD,CraigKDandMogilJS(2010)Codingoffacialexpressionsofpaininthelaboratorymouse.NatureMethods7,447–449.LazicSEandEssiouxL(2013)Improvingbasicandtranslationalsciencebyaccountingforlitter-to-littervariationinanimalmodels.BMCNeuroscience14,37.LeidigMS,HertrampfB,FailingK,SchumannAandReinerG(2009)Painanddiscomfortinmalepigletsduringsurgicalcastrationwithandwithout

chunk 42 · 365 tokens

models.BMCNeuroscience14,37.LeidigMS,HertrampfB,FailingK,SchumannAandReinerG(2009)Painanddiscomfortinmalepigletsduringsurgicalcastrationwithandwithout localanaesthesiaasdeterminedbyvocalisationanddefensebehavior.AppliedAnimalBehaviourScience116,174–178.LlamasMoyaS,BoyleLA,LynchPBandArkinsS(2008)Effectofsurgicalcastrationonthebehavioralandacutephaseresponsesof5-day-oldpiglets.AppliedAnimalBehaviourScience111,133–145.MagnaniD,GattoM,CafazzoS,StellettaC,MorganteMandCostaLN(2011)Differenceofsurfacebodytemperatureinpigletsduetothebacktestandenvironmentalcondition.InternationalCongressforAnimalHygiene,Vienna,Austria,3-7July2011,Volume3,pp.1029–1032.MalavasiLM,NymanG,AugustssonH,JacobsonMandJensen-WaernM(2006)Effectsofepiduralmorphineandtransdermalfentanylanalgesiaonphysiologyandbehaviourafterabdominalsurgeryinpigs.LaboratoryAnimals40,16–27.MarsálekP,SvobodaM,SmutnáM,BlahováJandVecerekV(2011)Neopterinandbiopterinasbiomarkersofimmunesystemactivationasso-ciatedwithcastrationinpiglets.JournalofAnimalScience89,1758–1762.MaršálekP,SvobodaM,BernardyJandVečerekV(2015)Concentrationsofneopterin,biopterin,andcortisolassociatedwithsurgicalcastrationofpig- letswithlidocaine.CzechJournalofAnimalScience60,473–478.MatthewKA(2000)Painassessmentandgeneralapproachtomanagement.TheVeterinaryClinicsofNorthAmerica.SmallAnimalPractice30,729–755.McGloneJJandHellmanJM(1988)Localandgeneralanestheticeffectsonbehaviorandperformanceoftwo-andseven-week-oldcastratedanduncas-

chunk 43 · 373 tokens

lAnimalPractice30,729–755.McGloneJJandHellmanJM(1988)Localandgeneralanestheticeffectsonbehaviorandperformanceoftwo-andseven-week-oldcastratedanduncas- tratedpiglets.JournalofAnimalScience66,3049–3058.McGloneJJ,NicholsonRI,HellmanJMandHerzogDN(1993)Thedevel-opmentofpaininyoungpigsassociatedwithcastrationandattemptstopreventcastration-inducedbehavioralchanges.JournalofAnimalScience71,1441–1446.McLennanKM,RebeloCJB,CorkeMJ,HolmesMA,LeachMCandConstantino-CasasF(2016)Developmentofafacialexpressionscaleusingfootrotandmastitisasmodelsofpaininsheep.AppliedAnimalBehaviourScience176,19–26.McMeekanCM,MellorDJ,StaffordKJ,BruceRA,WardRNandGregoryNG(1998)Effectsoflocalanaesthesiaof4or8hdurationontheacutecor-tisolresponsetoscoopdehorningincalves.AustralianVeterinaryJournal76,281–285.MeijerE,vanNesA,BackWandvanderStaayFJ(2015)Clinicaleffectsofbuprenorphineonopen-fieldbehaviourandgaitsymmetryinhealthyandlameweanedpiglets.VeterinaryJournal206,298–303.MellorDJandGregoryNG(2003)Responsiveness,behaviouralarousalandawarenessinfetalandnewbornlambs:experimental,practicalandthera-peuticimplications.NewZealandVeterinaryJournal51,2–13.MillerALandLeachMC(2015a)Themousegrimacescale:aclinicallyusefultool?PLoSOne10,e0136000.MillerALandLeachMC(2015b)Usingthemousegrimacescaletoassesspainassociatedwithroutineearnotchingandtheeffectofanalgesiainlaboratorymice.LaboratoryAnimals49,117–120.MillerA,KitsonG,SkalkoyannisBandLeachM(2015)Theeffectofisofluraneanaesthesiaandbuprenorphineonthemousegrimacescaleand

chunk 44 · 505 tokens

mice.LaboratoryAnimals49,117–120.MillerA,KitsonG,SkalkoyannisBandLeachM(2015)Theeffectofisofluraneanaesthesiaandbuprenorphineonthemousegrimacescaleand behaviourinCBAandDBA/2mice.AppliedAnimalBehaviourScience172,58–62.MillerAL,KitsonGL,SkalkoyannisB,FlecknellPAandLeachMC(2016)UsingthemousegrimacescaleandbehaviourtoassesspaininCBAmicefollowingvasectomy.AppliedAnimalBehaviourScience181,160–165.MoeRO,StubsjøenSM,BohlinJ,FløAandBakkenM(2012)Peripheraltemperaturedropinresponsetoanticipationandconsumptionofasignaledpalatablerewardinlayinghens(Gallusdomesticus).Physiology&Behavior106,527–533.MoralesJ,DereuA,MansoA,deFrutosL,PiñeiroC,ManzanillaEGandWuytsN(2017)Surgicalcastrationwithpainreliefaffectsthehealthandproductiveperformanceofpigsinthesucklingperiod.PorcineHealthManagement3,18.MosherRA,CoetzeeJF,AllenPS,HavelJA,GriffithGRandWangC(2014)EffectsofsamplehandlingmethodsonsubstancePconcentrationsandsub-stancePimmunoreactivityinbovineblood.AmericanJournalofVeterinaryResearch75,109–116.NasirahmadiA,EdwardsSA,MathesonSMandSturmB(2017)Usingauto-matedimageanalysisinpigbehaviouralresearch:assessmentoftheinflu-enceofenrichmentsubstrateprovisiononlyingbehaviour.AppliedAnimalBehaviourScience196,30–35.NationalFarmAnimalCareCouncil(2014)CodeofPracticefortheCareandHandlingofPigs.[Online]Availableat:https://www.nfacc.ca/codes-of-practice/pig-code#section4(accessed18/07/21).O’ConnorA,AnthonyR,BergamascoL,CoetzeeJ,GouldS,JohnsonAK,KarrikerLA,Marchant-FordeJN,MartineauGS,McKeanJ,MillmanST,NiekampS,PajorEA,RutherfordK,SpragueM,SutherlandM,vonBorellEandDzikamunhengaRS(2014)Painmanagementintheneonatalpigletduringroutinemanagementprocedures.Part2:gradingthequalityofevidenceandthestrengthofrecommendations.AnimalHealthResearchReviews15,39–62.O’ConnorA,AnthonyR,BergamascoL,CoetzeeJF,DzikamunhengaRS,JohnsonAK,KarrikerLA,Marchant-FordeJN,MartineauGP,MillmanST,PajorEA,RutherfordK,SpragueM,SutherlandMA,vonBorrellEandWebbSR(2016)Review:assessmentofcompletenessofreportingininterventionstudiesusinglivestock:anexamplefrompain

chunk 45 · 197 tokens

rfordK,SpragueM,SutherlandMA,vonBorrellEandWebbSR(2016)Review:assessmentofcompletenessofreportingininterventionstudiesusinglivestock:anexamplefrompain mitigationinterventionsinneonatalpiglets.Anim10,660–670.Pairis-GarciaMD,JohnsonAK,AbellCA,CoetzeeJF,KarrikerLA,MillmanSTandStalderKJ(2015)MeasuringtheefficacyofflunixinmeglumineandmeloxicamforlamesowsusingaGAITFourpressurematandanembeddedmicrocomputer-basedforceplatesystem.JournalofAnimalScience93,2100–2110.ParkR,CramerM,WagnerB,TurnerP,MoraesL,ViscardiA,CoetzeeJandPairis-GarciaM(2020)Acomparisonofbehaviouralmethodologiesutilisedtoquantifydeviationsinpigletbehaviourassociatedwithcastration.AnimalWelfare29,285–292.AnimalHealthResearchReviews175 https://doi.org/10.1017/S1466252321000141 Published online by Cambridge University Press

chunk 46 · 227 tokens

PrunierA,MounierAMandHayM(2005)Effectsofcastration,toothresec-tion,ortaildockingonplasmametabolitesandstresshormonesinyoungpigs.JournalofAnimalScience83,216–222.PrunierA,BonneauM,vonBorellEH,CinottiS,GunnM,FredriksenB,GiersingM,MortonDB,TuyttensFAMandVelardeA(2006)Areviewofthewelfareconsequencesofsurgicalcastrationinpigletsandtheevalu-ationofnon-surgicalmethods.AnimalWelfare15,277–289.RaultJL,LayDCandMarchant-FordeJN(2011)Castration-inducedpaininpigsandotherlivestock.https://www.ars.usda.gov/ARSUserFiles/50201500/Castration%20Fact%20Sheet.pdf.USDA-ARS-MWAFarmAnimalWelfareFactSheet.PublishedJune2011.AccessedDecember7,2018.ReimertI,FongS,RodenburgTBandBolhuisJE(2017)Emotionalstatesandemotionalcontagioninpigsafterexposuretoaandnegativetreatment.AppliedAnimalBehaviourScience193,37–42.RossL,CressmanM,CramerMandPairis-GarciaM(2019)Validationofalternativebehavioralobservationmethodsinyoungbroilerchickens.

chunk 47 · 355 tokens

imalBehaviourScience193,37–42.RossL,CressmanM,CramerMandPairis-GarciaM(2019)Validationofalternativebehavioralobservationmethodsinyoungbroilerchickens. PoultryScience98,6225–6231.SchaeferAL,AalhusJL,CookNJ,DuganMER,DubeskiPL,FortinAF,RobertsonWMandTongAKW(2002)Pigwelfare:measuringstressinmarketpigs:treatingstressinmarketpigs.AdvancesPorkProduction13,29.SchmidtT,KönigAandvonBorellE(2012)Impactofgeneralinjectionanaesthesiaandanalgesiaonpost-castrationbehaviorandteatorderofpig-lets.Animal:AnInternationalJournalofAnimalBioscience6,1998–2002.SchulzKL,AndersonDE,CoetzeeJF,WhiteBJandMiesnerMD(2011)EffectofflunixinmeglumineontheameliorationoflamenessindairysteerswithamphotericinB-inducedtransientsynovitis-arthritis.AmericanJournalofVeterinaryResearch72,1431–1438.SheilMandPolkinghorneA(2020)Optimalmethodsofdocumentinganal-gesicefficacyinneonatalpigletsundergoingcastration.Animals10,1450.SneddonL,ElwoodR,AdamoSandLeachM(2014)Definingandassessinganimalpain.AnimalBehaviour97,201–212.h.SotocinalSG,SorgeRE,ZaloumA,TuttleAH,MartinLJ,WieskopfJS,MapplebeckJCS,WeiP,ZhanS,ZhangS,McDougallJJ,KingODandMogilJS(2011)Theratgrimacescale:apartiallyautomatedmethodforquantifyingpaininthelaboratoryratviafacialexpressions.MolecularPain7,55.StaffordKJ,MellorDJ,ToddSE,BruceRAandWardRN(2002)Effectsoflocalanaesthesiaorlocalanaesthesiaplusnon-steroidalanti-inflammatorydrugintheacutecortisolresponseofcalvestofivedifferentmethodsofcas-

chunk 48 · 451 tokens

N(2002)Effectsoflocalanaesthesiaorlocalanaesthesiaplusnon-steroidalanti-inflammatorydrugintheacutecortisolresponseofcalvestofivedifferentmethodsofcas- tration.ResearchinVeterinaryScience73,61–70.StewartM,WebsterJR,SchaeferAL,CookNJandScottSL(2005)Infraredthermographyasanon-invasivetooltostudyanimalwelfare.AnimalWelfare14,319–325.StewartM,VerkerkGA,StaffordKJ,SchaeferALandWebsterJR(2010)Non-invasiveassessmentofautomaticactivityforevaluationofpainincalves,usingsurgicalcastrationasamodel.JournalofDairyScience93,3602–3609.SutherlandMA,MellorDJ,StaffordKJ,GregoryNG,BruceRA,WardRNandToddSE(1999)Acutecortisolresponsesoflambstoringcastrationanddockingaftertheinjectionoflignocaineintothescrotalneckortestesatthetimeofringapplication.AustralianVeterinaryJournal77,738–741.SutherlandMA,MellorDJ,StaffordKJ,GregoryNG,BruceRAandWardRN(2002)Cortisolresponsestodehorningofcalvesgivena5-hourlocalanestheticregimenplusphenylbutazone,ketoprofenoradrenocorticotropichormonepriortodehorning.ResearchinVeterinaryScience73,115–123.SutherlandMA,BryerPJ,KrebsNandMcGloneJJ(2008)Taildockinginpigs:acutephysiologicalandbehavioralresponses.Animal:AnInternationalJournalofAnimalBioscience2,292–297.SutherlandMA,DavisBL,BrooksTAandMcGloneJJ(2010)Physiologyandbehaviorofpigsbeforeandaftercastration:effectsoftwotopicalanesthetics.Animal:AnInternationalJournalofAnimalBioscience4,2071–2079.SutherlandMA,DavisBL,BrooksTAandCoetzeeJF(2012)Thephysio-logicalandbehavioralresponseofpigscastratedwithandwithoutanesthe-siaand/oranalgesia.JournalofAnimalScience90,2211–2221.SutherlandM,DowlingS,BackusBandStewartM(2015)Theuseofinfra-redthermographytoassessstressinpigs.Proc7thEurConfPrecisLivestFarming.691–699.SutherlandMA,BackusBL,BrooksTAandMcGloneJJ(2017)Theeffectofneedle-freeadministrationoflocalanestheticonthebehaviorandphysi-

chunk 49 · 204 tokens

recisLivestFarming.691–699.SutherlandMA,BackusBL,BrooksTAandMcGloneJJ(2017)Theeffectofneedle-freeadministrationoflocalanestheticonthebehaviorandphysi- ologyofcastratedpigs.JournalofVeterinaryBehavior21,71–76.TorrisonJ(2012)Optimizingdiagnosticvalueandsamplecollection.InZimmermanJJ,KarrikerLA,RamirezA,SchwartzKJandStevensonGW(eds.),DiseasesofSwine.10thEdn.Ames,IA:JohnWiley&Sons,pp.67–76.TuyttensFAM,VanhonackerF,LangendriesK,AluweM,MilletBekaertKandVerbekeW(2011)Effectofinformationprovisioningonattitudetowardsurgicalcastrationofmalepigletsandalternativestrategiesforavoidingboartaint.ResearchinVeterinaryScience91,327–332.USFoodandDrugAdministration(2017)Banamine®Transdermalflunixintransdermalsolution.Steers,beefheifers,beefcows,beefbullsintendedforslaughter,andreplacementdairyheifersunder20monthsofage.Forthe

chunk 50 · 523 tokens

e®Transdermalflunixintransdermalsolution.Steers,beefheifers,beefcows,beefbullsintendedforslaughter,andreplacementdairyheifersunder20monthsofage.Forthe controlofpyrexiaassociatedwithbovinerespiratorydiseaseandthecontrolofpainassociatedwithfootrot.https://animaldrugsatfda.fda.gov/adafda/app/search/public/document/downloadFoi/1944.FreedomofInformationSummaryNADA-141-450.PublishedJuly21,2017.AccessedNovember7,2018.USFoodandDrugAdministration-CenterforVeterinaryMedicine(2006)Developmentoftargetanimalsafetyandeffectivenessdatatosupportapprovalofnon-steroidalanti-inflammatorydrugs(NSAIDS)foruseinanimals.http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/GuidanceforIndustry/UCM052663.pdf.GuidanceforIndustryNo.123.PublishedJanuary5,2006.AccessedNovember7,2018.ViscardiAVandTurnerPV(2018a)Efficacyofbuprenorphineformanage-mentofsurgicalcastrationpaininpiglets.BMCVeterinaryResearch14,318.ViscardiAVandTurnerPV(2018b)Useofmeloxicamandketoprofenforpigletpaincontrolfollowingsurgicalcastration.FrontiersinVeterinaryScience5,299.ViscardiAV,HunnifordM,LawlisP,LeachMandTurnerPV(2017)Developmentofapigletgrimacescaletoevaluatepigletpainusingfacialexpressionsfollowingcastrationandtaildocking:apilotstudy.FrontiersinVeterinaryScience4,51.WearyDM,BraithwaiteLAandFraserD(1998)Vocalresponsetopaininpiglets.AppliedAnimalBehaviourScience56,161–172.WearyDM,NielL,FlowerFCandFraserD(2006)Identifyingandprevent-ingpaininanimals.AppliedAnimalBehaviourScience100,64–76.WhiteRG,DeShazerJA,TresslerCJ,BorcherGM,DaveyS,WaningeA,ParkhurstAM,MilanukMJandClemensET(1995)Vocalizationandphysiologicalresponseofpigsduringcastrationwithorwithoutalocalanesthetic.JournalofAnimalScience73,381–386.YunJ,OllilaA,ValrosA,Larenza-MenziesP,HeinonenM,OlivieroCandPeltoniemiO(2019)Behaviouralalterationsinpigletsaftersurgicalcastra-tion:effectsofanalgesiaandanaesthesia.ResearchinVeterinaryScience125,36–42.ZanklA,RitzmannM,ZoilsSandHeinritziK(2007)Theefficacyoflocalanaestheticsadministeredpriortocastrationofmalesucklingpiglets.DTW.DeutscheTierarztlicheWochenschrift114,418–422.176AngelaBaysingeretal.

chunk 51 · 60 tokens

fficacyoflocalanaestheticsadministeredpriortocastrationofmalesucklingpiglets.DTW.DeutscheTierarztlicheWochenschrift114,418–422.176AngelaBaysingeretal. https://doi.org/10.1017/S1466252321000141 Published online by Cambridge University Press