Lomax Sheil Windsor_2013_AVJ

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DurationofactionofatopicalanaestheticformulationforpainmanagementofmulesinginsheepSLomax,a*MSheilbandPAWindsora ObjectiveToinvestigatetheeffectoftopicalanaesthesiaon‘motheringup’oflambsaftermulesingandmarking,andforpain alleviationovera24-hperiod.DesignTwoseparatetrialswereperformedonMerinolambsundergoingthemulesprocedureforflystrikeprevention,to assesstheefficacyofimmediatepostoperativetopicalanaesthetic wounddressingcontaininglignocaine(hydrochloride)40.6g/L, bupivacaine(hydrochloride)4.5g/L,adrenaline(tartrate)24.8mg/L andcetrimide5.0g/Linagelbase(BayerAnimalHealth,Gordon, NSW,Australia).MethodsInbothtrials,lambswereassignedtooneofthreetreat-mentregimens:control,mulesprocedurewithtopicalanaesthetic (0.5mL/kg)andmulesprocedurewithouttopicalanaesthetictreat- ment.Parametersmeasuredincludedbodyweight,assessmentof skinandwoundsensitivitytolighttouchandpainstimulation, behaviouralresponsesandtimetomotherupandtofeed.ResultsInbothtrialstherewasrapid(1min)andprolonged(upto24h)woundanalgesiaasshownbylowerscoresforlighttouch (P<0.001)andpainresponses(P<0.001),withabsentorsignifi-cantlydiminishedprimaryandsecondaryhyperalgesia(P 0.05)andsignificantreductioninpain-relatedbehaviours(P<0.001)intreatedversusuntreatedlambs.ConclusionSignificantpainalleviationandimprovedrecoverycanbeachievedinlambsforatleast24haftermulesingthrough theuseoftopicalanaesthesia.Itissuggestedthatthehaemostatic actionofadrenalin,togetherwithinhibitionoftheinflammatory cascadeandthebarriereffectofthegelwithintheproduct,may explaintheprolongedanaesthesiaupto24hobservedinthe presentstudy.Theseresultssuggestthattopicalanaesthesiahas thecapacitytodramaticallyimprovethewelfareoflambsunder- goingmulesing.

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aupto24hobservedinthe presentstudy.Theseresultssuggestthattopicalanaesthesiahas thecapacitytodramaticallyimprovethewelfareoflambsunder- goingmulesing. Keywordsallodynia;flystrike;hyperalgesia;mulesing;sheep;vonFreymonofilamentAbbreviationsHI,hotiron;LT,lighttouch;NRS,numericalratingscale;NSAID,non-steroidalantiinflammatorydrugAustVetJ2013;91:160–167doi:10.1111/avj.12031 F lystrikeisoneofthemostsignificanthealthandwelfarecon-cernsfortheAustralianandNewZealandsheepindustries.Itis adiseaseresultingfromtheinfestationoflivingtissuebyflylarvae,initiatedbyovipositionbythesheepblowflyLuciliacuprina.Breechstrikeinvolvinginfestationofthe‘crutch’orperinealregionisthemostcommonformofblowflystrike,occurringtosomedegreein almosteverysusceptibleflockeachyearinAustralia.1–4Themulesprocedure,developedinthe1930s,involvestheremovalof thefoldsofskinfromthebreechareaofsheepinordertodecrease susceptibilitytoovipositionbyL.cuprinathroughreducedmoistureandfaecesretention.Itiswidelyacceptedasthemosteffectivemethod forthelifetimepreventionofbreechstrike.5Despitethepreventivehealthbenefits,therehasbeengrowing concernregardingthewelfareimpactoftheprocedureitself.Inthe past,theprocedurewasundertakenwithoutanalgesia,resultingin evidenceofacutepainandstress.6–9TheAustralianWoolIndustryhasfacedincreasingpressureinrecentyearstodevelopalternative,more humanemethodsofflystrikepreventiontosupportaphase-outofthe mulesprocedure.Theultimatesolutionisbelievedtolieinbreeding sheepresistanttobreechstrike(suchthatmulesingisnolonger required).Intensivegeneticresearchandbreedingprogrammesare underway,butthisisalong-termobjective.10–12Additionalresearchistargetingthemoreimmediategoalofdevelopingmorehumaneprac-

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searchandbreedingprogrammesare underway,butthisisalong-termobjective.10–12Additionalresearchistargetingthemoreimmediategoalofdevelopingmorehumaneprac- ticestodealwithbreechwrinkle.In2005inAustralia,amultifunctiontopicalanaesthetic,antisepticandhaemostaticwounddressing(Tri-Solfen®,BayerAnimalHealth,Gordon,NSW,Australia)becameavailableforuseonpermitthroughveterinariansforimmediatepost-proceduralapplicationtomulesingwoundsinsheep.Tri-Solfen®isaspray-ontopicalanaesthetic,hae-mostaticandantisepticgelagentconsistingoflignocaine(40.6g/L),bupivacaine(4.5g/L),adrenalin(24.8mg/L)andcetrimide(5.0g/L). Wehavebeenexaminingtheefficacyofthisformoftreatmenttoboth alleviatepainandenhancewoundhealingandrecoveryinthefirst8h aftertheprocedure.Ourstudiesindicatethatsignificantwound desensitisation,improvedlambrecoveryratesandenhancedwound healingcanbeachieved.13Observationsduringourstudies,togetherwithanecdotalreportsof improvedlambbehaviourupto24hpost-mulesing,ledustohypoth- esisethattreatmentwiththetopicalanaestheticproductmayeffecta prolongedanalgesicresponsebecauseofinhibitionofthepainesca- lationresponse.Themother–lambbondisimportanttolambsurvivalearlyinlifeanditcanbedisruptedbyhumaninterventions,particularlyin Merinoewes,whichhavethereputationofbeingpoormothers.14Incommercialfarmsituations,lambsarereturnedimmediatelyto pasturewiththeirmothersfollowinghusbandryproceduressuchas mulesing.Thisprocessisreferredtoas‘motheringup’andisimpor- tantforimprovedlambrecoverybecausethefirstfeedofmilkleadsto ariseinendorphins,whichcanameliorateacutepain.Additionally, *Correspondingauthor.aFacultyofVeterinaryScience,UniversityofSydney,PMB3,Camden,NewSouthWales2570,Australia; sabrina.lomax@sydney.edu.au bAnimalEthicsPtyLtd,YarraGlen,Victoria,Australia bs_bs_banner

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,UniversityofSydney,PMB3,Camden,NewSouthWales2570,Australia; sabrina.lomax@sydney.edu.au bAnimalEthicsPtyLtd,YarraGlen,Victoria,Australia bs_bs_banner PRODUCTIONANIMALSPRODUCTIONANIMALS ©2013TheAuthorsAustralianVeterinaryJournal©2013AustralianVeterinaryAssociationAustralianVeterinaryJournalVolume91,No4,April2013160

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lambsthathaveadelayinthetimetomotheruparemoresusceptibletoexposure,includinghypothermiaanddehydration,whichcanlead tomorbidityandinsomecasesmortality.14Therearenodiscretedataintheliteraturedescribingtheeffectsofmulesingonthetimeto motherupandfeedinlambs,despitethepainfulnatureoftheproce- dure.Thisinformationhasthepotentialtoprovideuswithasimple tooltoassesslambdiscomfortanddeterminetheefficacyofpain managementregimensforimprovinglambrecoverypost-mulesing.Inthepresentstudy,weexaminedtheeffectofmulesingonmotheringupoflambsfollowingmulesingandmarking,andthedurationof efficacyofatopicalanaestheticforimprovinglambrecoveryand alleviatingpainovera24-hperiod.MaterialsandmethodsTwotrialswereperformedon6–8-week-oldMerinolambsfromacommercialflockintheSouthernHighlands,NewSouthWales,Aus- tralia.Allanimalprocedureswereconductedwithpriorinstitutional animalethicsapprovalinaccordancewiththeNationalHealthand MedicalResearchCouncil’sCodeofPracticefortheCareandUseofAnimalsforScientificPurposes.ThetrialswereconductedinJuly2008(Trial1;n=23)onlambsundergoingbothmulesingandhot-iron(HI)taildockingandinJanuary2009(Trial2;n=42)onlambsundergoingmulesingonly(previouslytaildocked).Themeaninitial bodyweightoflambsinTrial1was11.89 0.3kgandforlambsinTrial2itwas21.04 0.5kg.Inbothtrials,lambswereassignedtooneofthefollowingtreatmentregimens,randomisedwithineachtrial (Table1):(1)control,inwhichlambswerehandledbutremained unmulesed(ortaildockedinTrial1);(2)mulesing(andHItail dockinginTrial1)withimmediatepostoperativetopicalanaestheticwounddressing(6–12mLofTri-Solfen®sprayedontowound);(3)mulesing(withHItaildockinginTrial1)withouttopicalanaestheticdressing;and(4)HItaildockingonly(Trial1,behaviouralobserva-

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ri-Solfen®sprayedontowound);(3)mulesing(withHItaildockinginTrial1)withouttopicalanaestheticdressing;and(4)HItaildockingonly(Trial1,behaviouralobserva- tionsasdescribedlater).GeneralmanagementandmarkingOnthedayofeachtrial,lambswereyardedanddraftedfromtheirdamsintoaholdingyardwheretheywereheldseparatelyfromtheirdamsfor4h.Theywerethenselectedatrandom,weighedandspray-markednumericallyandplacedinmarkingcradles.Preoperative skinsensitivitytolighttouch(LT)andpainstimulationwasrecorded (seelater).Mulesingwasperformedusingastandard‘V’-modified technique.InTrial1,theHItaildockingwasperformedusingagas tail-dockingknifethathadbeenpreheatedtothecorrecttemperature toefficientlysealthecoccygealbloodvessels.Theskinofthetailwas pushedtowardsthelamb’sbodytolocatethecorrectpositionbetween coccygealvertebrae2and3andthetailelevatedtoavoidburningof theperineum.Theleveroftheknifewassqueezedandafter2sthetailwasremoved.Inlambsassignedtotreatmentgroup2,Tri-Solfen®wasapplieddirectlytothemulesingandtail-dockingwoundsusingametereddoseapplicator(6–12mL,basedonlambweight)immedi- atelyaftertheprocedure(s).Postoperativeskinandwoundsensitivity testingwasrepeatedat1minpost-procedure.Lambswerereturnedto theirdamsina6¥4mpen,wherethetimetakenforlambstofindtheirdamsandfeedwasrecorded.Theewesandlambswerethen movedtooneofthreepasture-coveredholdingyards(20¥10m)inmixedtreatmentgroupsandleftundisturbedfor24h.At1hand 24hpost-mulesing,thebehaviouralresponsescoreswererecorded (Trial1)asdescribedlater.At24hafterthebehaviouralassessment (Trial1),lambswerethenre-drafted,weighedandskinandwound

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aviouralresponsescoreswererecorded (Trial1)asdescribedlater.At24hafterthebehaviouralassessment (Trial1),lambswerethenre-drafted,weighedandskinandwound sensitivitytestingwasrepeated.BodyweightsWeightswererecordedusingdigitalscales(Rudweigh®,GallagherAnimalManagementSystems,Australia),whichwerecalibratedandzeroedpriortoeachmeasurementandaccurateto0.1kg.Assessmentofskinandwoundsensitivitytolighttouchandpainstimulation Skinandwoundsensitivitywasassessedatfivepredeterminedsitesontheskinsurroundingthemulesingwoundandninesiteswithinthe mulesingwoundbeforeandthen1minand24haftermulesingusing weightedvonFreymonofilaments,whicharecalibratedtobendat predeterminedpressurestoproviderepeatableLT(10N)andpain (75N)stimulationofthewound,aspreviouslydescribed.13Evidenceoflocalanaesthesia,primary(wound)andsecondary (tissuesurroundingwound)allodynia(painresponsetonon-painful stimuli)andhyperalgesia(exaggeratedresponsetopainfulstimuli) wereassessedateachsite.TypicalLT-andpain-inducedinvoluntary reflexesandmotorresponsesintherumpandheadweremeasured usingacustomisednumericalratingscale(NRS).13Rumpresponsescoresweregradedas:0,noresponse;1,minorinvoluntarymotor responsesuchaslocalskintwitch,subcutaneousmuscletwitchoranal contraction;2,partialrumpwithdrawalreflexsuchasmultiplesub- cutaneousmusclegroupcontractionand/orliftingofthetail;3,full rumpwithdrawalreflexwithliftingoftherumpoffthecradle.Facial responsescoresweregradedas:0,noresponse;1,minorfacial‘aware- ness’suchaseyewidening,blinkingornasalflaring;2,partialstartle reflexoftheheadsuchasslightliftingofthesnoutorpartialhead rotation;3,fullstartlereflexofthehead,resultinginamajormove- mentsuchasliftingtheheadoffthecradle,fullheadjerkorfullhead rotation.Scoresforeachsitewereaddedtoachieveatotalscorefor

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ofthehead,resultinginamajormove- mentsuchasliftingtheheadoffthecradle,fullheadjerkorfullhead rotation.Scoresforeachsitewereaddedtoachieveatotalscorefor eachlamb.Totalscoreswerecalculatedoutof30forskinsensitivity aroundthemulesedareaandoutof54fordirectwoundsensitivity. Table1. TreatmentgroupsandtotalnumberoflambsineachtreatmentgroupinTrials1and2 GroupNo.oflambsMulesingTaildockingTreatmentTrial115NoNoControl 26YesHotironTopicalanaesthetic 36YesHotironNoanaesthetic 46NoHotironNoanaesthetic Trial2 114NoNoControl 214YesNoTopicalanaesthetic 314YesNoNoanaesthetic PRODUCTIONANIMALSPRODUCTIONANIMALS ©2013TheAuthorsAustralianVeterinaryJournal©2013AustralianVeterinaryAssociationAustralianVeterinaryJournalVolume91,No4,April2013161

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MeasurementoftimestomotherupandtofeedLambsaged6–8weeksfrom3-year-oldeweswereusedforthesetrials.Ineachtrial,lambsandmothers(dams)wereobservedingroupsof mixedtreatment.Lambsanddamswereidentifiedinpairsbycol- ouredmarkerforobservationoftimestomotherupandtofeed. InTrial1,therewerethreegroupsof7–8lambsanddamsandinTrial 2,sixgroupsof7.Therewerenotwinsincludedinthetrial.Time(s)takenforindividuallambstofindtheirmother,andtofeedwererecordedusingastopwatch(DSE®digitalLCD).Timetomotherupwasmeasuredastimetakenforthelambtolocateandberecog-nisedbyitsmotherfromitstimeofreleaseintotheyard.Timetofeed wasmeasuredasthetimetakenforthelambtobeginsucklingfrom theewefromitstimeofreleasefromtheyard.Thisdidnotalways happensimultaneouslywithmotheringup.DuringTrial1,weobservedthatseveralewesdisplayedcirclingbehav-iourwhenlambsattemptedtofeed,delayingthetimetofeedresponse. WepostulatedthattheodourfromsingedwoolassociatedwiththeHI taildockingwoundmighthavebeenconfusingtheewesandthus confoundingtheresponse.Toinvestigatethisfurther,afourthgroup oflambsthatunderwentHItaildockingonly(n=6,nomulesing,andnotopicalanaesthetictreatment)wasincludedtoassessthetimesto motherupandtofeed.Theselambsunderwentthesametreatment sequenceandwerereleasedindividuallyintothepenofthesixdams immediatelyfollowingtreatment.Assessmentofpain-relatedbehavioursInTrial1,pain-relatedbehaviourwasassessedat1hand24hpost-mulesingusingtheNRSdevelopedpreviously.13Atrainedobserver(experiencedanimalscientist‘blindtotreatment’)observedthelambs 24hpost-procedure.Individuallambsweregivenascorebetween 0and3:0,nopain-relatedbehaviour;1,mildabnormalitiesofposture, gaitorbehavioursuchasmildkyphosiswithouthyperextensionof

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duallambsweregivenascorebetween 0and3:0,nopain-relatedbehaviour;1,mildabnormalitiesofposture, gaitorbehavioursuchasmildkyphosiswithouthyperextensionof hindlegs,ventralrecumbencywithhindlegspartiallyextendedormildstiffeningofgaitwithoutovertlimpingorlegdragging;2,moderate abnormalitiesofposture,gaitorbehavioursuchas‘statuestanding’ withheaddownandprominentkyphosis,moderatestiffeningor slowingofgaitorhyperextensionand/orabductionofhindlegs, ventralrecumbencywithhindlegsfullyextended;3,displayofsevere abnormalitiesofposture,gaitorbehavioursuchasmarkedagitation withtwistingorwrithing,highfrequencyofposturalchangefrom lyingtokneelingorstanding,distressedvocalisation,lateralorpros- tratelying,kneeling,dogsittingortremors,shakingorlipcurling.StatisticalanalysisDatawereanalysedusingGenStat®version10.0(VSNInternationalLtd,HemelHempstead,UK).One-wayanalysisofvariancewasusedtomeasureeffectsoftreatmentontimestomotherupandtofeed. Dataweretestedfornormalityandwherenecessarytransformedinto alogarithmformtonormalisedistribution.Arepeatedmeasures residualmaximumlikelihoodestimationforlinearmixedmodelswas usedtoanalyseweightdataandNRSscoresfromsensitivitytesting andbehaviouralobservations.Foralltreatments,whereasignificant timeandtreatmentinteractionwasfound,post-hocpair-wise

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aandNRSscoresfromsensitivitytesting andbehaviouralobservations.Foralltreatments,whereasignificant timeandtreatmentinteractionwasfound,post-hocpair-wise comparisonsusingleastsignificantdifferenceswereperformedtocomparedifferenceswithintime-points.Forallstatisticalcalculations,P<0.05wasconsideredstatisticallysignificant.ResultsBodyweightsTrial1.Thechangeinbodyweightfrompretreatment(11.86 0.32kg)to24hpost-treatment(12.26 0.32kg)wassignificant(P=0.03).Treatment(P=0.59)andtreatmentbytimeinteraction(P=0.297)werenotsignificant.Trial2.Thechangeinbodyweightfrompretreatment(21.05 0.49kg)to24hpost-treatment(20.48 0.45kg)wasnotsignificant(P=0.404).Treatment(P=0.737)andtreatmentbytimeinteraction(P=0.999)werenotsignificant.TimestomotherupandtofeedTrial1.ResultsfromTrial1aredisplayedinFigure1.LambsthathadbeenHItaildockedwithorwithoutmulesingand/ortreatmentwithtopicalanaestheticdressinghadsignificantly(P<0.05)longermeantimestomotherupandtofeedthanundocked/unmulesedcontrol lambs.Behaviourofewes,includingcirclingoflambsandsmellingof theperineum,wasnotedwhenlambswereplacedinthepenwith dams.Therewasnosignificantdifferenceintimetomotherup (P 0.13)ortimetofirstfeed(P 0.12)betweenanyofthegroupsoflambsthatwereHItaildocked,regardlessofmulesingtreatment.Trial2.ResultsfromTrial2aredisplayedinFigure2.Lambsinthisgrouphadbeenpreviouslytaildockedandthusunderwentmulesingonly.Fieldobservationsindicatedthatdamsdidnotdisplaythecir- clingavoidancebehaviourthathadcharacterisedobservationsinthe Trial1groupsoflambsthathadundergoneHItaildocking.Thiswas reflectedinthetimetomotherup,whichdidnotdiffersignificantly betweenthegroupsandwassimilartothecontrolgroupfromTrial1. However,therewasatrendtowardsatreatmenteffectontimetofeed,

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herup,whichdidnotdiffersignificantly betweenthegroupsandwassimilartothecontrolgroupfromTrial1. However,therewasatrendtowardsatreatmenteffectontimetofeed, althoughnotsignificant(P=0.1).Mulesed,non-anaesthetisedlambstooklongertobeginsuckling(38s)thaneithercontrollambs(22s)or mulesedlambstreatedwithtopicalanaesthesia(12s). Figure1. Mean SEMtimestakenforlambsinTrial1tomotherupandtofeedimmediatelyaftertreatment.HI,hotiron;TA,topicalanaesthesia (Tri-Solfen®);NA,noanaesthesia. PRODUCTIONANIMALSPRODUCTIONANIMALS ©2013TheAuthorsAustralianVeterinaryJournal©2013AustralianVeterinaryAssociationAustralianVeterinaryJournalVolume91,No4,April2013162

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ResponsetolighttouchandpainstimulationofthewoundandsurroundingskinPre-mulesing.TherewasverylittletonoresponsetoLTorpainstimulationoftheintactskinofthebreechpriortomulesing.MeanresponsescoresfromthefiveLTtestingsites(maximumpossible scoreof30)andninepainsites(maximumpossiblescoreof54)were 0.04 0.04and 0.75 0.22,respectively,acrossbothtrials.Therewerenosignificantdifferencesbetweengroupswithineachtrial.Post-mulesing.MeanresponsestopainandLTstimulationofthewoundandperi-woundinTrial1areshowninFigures3and4,respec-tively.Therewassignificanteffectoftreatmentonresponsestopain andLTstimulationofthewound(bothP<0.001)andperi-wound(bothP<0.001).AsignificanttimebytreatmentinteractionwasalsoseenforLTandpainstimulationofthewound(bothP<0.001)andpainstimulationoftheperi-wound(P<0.001).Primaryhyperalgesia Figure2. Mean SEMtimestakenforlambsinTrial2tomotherupandtofeedimmediatelyaftertreatment.LambsinTrial2hadbeentail dockedpriortomulesing.TA,topicalanaesthesia(Tri-Solfen®);NA,noanaesthesia. Figure3. Mean SEMtotalheadandrumpresponsescorestopainstimulationofthe woundandperi-woundsurfacesbefore (0min)andaftertreatment(Trial1).TA, topicalanaesthesia(Tri-Solfen®);NA,noanaesthesia. Figure4. Mean SEMtotalheadandrumpresponsescorestolighttouchstimulationof thewoundandperi-woundsurfacesbefore (0min)andaftertreatment(Trial1).TA, topicalanaesthesia(Tri-Solfen®);NA,noanaesthesia. PRODUCTIONANIMALSPRODUCTIONANIMALS ©2013TheAuthorsAustralianVeterinaryJournal©2013AustralianVeterinaryAssociationAustralianVeterinaryJournalVolume91,No4,April2013163

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andallodynia(ofthewound)developedwithin1minofmulesinginnon-anaesthetisedlambs,butnotinlambsinthecontrolgroupor mulesedlambsthatreceivedtopicalanaesthesia.Mulesedlambsthat receivedtopicalanaesthesiaalsoexhibitedsignificantlylesssecondary hyperalgesia(P<0.001)andallodynia(P=0.05)thanmulesed,non-anaesthetisedlambs.Responsescoresofmulesed,non-anaesthetised lambstoLTandpainstimulationofthemulesingwoundwere significantlyhigherthanthoseofthecontrollambsandmulesed lambsthatreceivedtopicalanaesthesia,atboth1minand24h post-mulesing.MeanresponsestopainandLTstimulationofthewoundandperi-woundinTrial2areshowninFigures5and6,respectively.Therewas asignificanteffectoftreatment(P<0.001)andtimebytreatmentinteractioninresponsestopainstimulationofthemulesingwound (P<0.001)andsurroundingskin(P=0.016).Primaryandsecon-daryhyperalgesiadevelopedwithin1minofmulesinginnon-anaesthetisedlambs,butnotincontrollambsandmulesedlambsthatreceivedtopicalanaesthesia.Mulesedlambstreatedwithtopical anaesthesiahadsignificantlylowerresponsescorestopainstimula- tionofthemulesingwound(P<0.001)andsurroundingskin(P=0.016)at1minand24hpost-mulesingthanmulesed,non-anaesthetisedlambs.Primaryandsecondaryhyperalgesiaresponse scoresofmulesedlambsthatreceivedtopicalanaesthesiadidnot differsignificantlyfromcontrolsuntil24hpost-mulesing.Therewas nosignificantdifferenceinprimaryorsecondaryallodyniafound betweentreatmentgroups,despiteatrendformulesed,non- anaesthetisedlambstohavehigherNRSscoresthanmulesedlambs thatreceivedtopicalanaesthesiaandcontrollambs.Pain-relatedbehaviourTrial1.BehaviouraldataareshowninFigure7.Therewasasignifi-canteffectoftreatment(P<0.001),butnosignificanttreatmentbytimeinteraction.Mulesedlambstreatedwithtopicalanaesthetic Figure5. Mean SEMtotalheadandrumpresponsescorestopainstimulationofthe

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nosignificanttreatmentbytimeinteraction.Mulesedlambstreatedwithtopicalanaesthetic Figure5. Mean SEMtotalheadandrumpresponsescorestopainstimulationofthe woundandperi-woundsurfacesbefore (0min)andaftertreatment(Trial2).TA, topicalanaesthesia(Tri-Solfen®);NA,noanaesthesia. Figure6. Mean SEMtotalheadandrumpresponsescorestolighttouchstimulationof thewoundandperi-woundsurfacesbefore (0min)andaftertreatment(Trial2).TA, topicalanaesthesia(Tri-Solfen®);NA,noanaesthesia. PRODUCTIONANIMALSPRODUCTIONANIMALS ©2013TheAuthorsAustralianVeterinaryJournal©2013AustralianVeterinaryAssociationAustralianVeterinaryJournalVolume91,No4,April2013164

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displayedsignificantlylowerpain-relatedbehaviourscorescomparedwithmulesed,non-anaesthetisedlambsatboth1hand24hpost- mulesing.Mulesedlambstreatedwithtopicalanaestheticdidnot differsignificantlyfromunmulesedcontrols.DiscussionResultsfromourtrialsindicatethatsignificantpainalleviationandimprovedrecoverycanbeachievedinlambsforupto24hafter mulesingthroughtheuseoftopicalanaesthesia.Thisextendsdata frompreviousresearchthattopicalanaesthesiaisefficaciousupto 8hpost-mulesing.13,15InTrial1,HItaildockingseemstohavebeenthemajorfactordelaying thetimestomotherupandtofeed.Thiscouldbeattributedtothe invasivenatureoftheprocedure,asitiswelldocumentedthattail dockingelicitsasignificantpainresponseinlambs.9,16–18Itisalsohypothesisedthattheodourofthecauterisedwoundmayactasa deterrenttothemotherwhenlambsapproachthemandthiswas observedthroughtheavoidancebehaviourdisplayedbytheewesto thelambs,evidentinTrial1,butnotinTrial2wherelambswere mulesedwithouttaildocking.EwesinTrial1appearedtospendmore timesmellingthelambs,particularlytheperineum,beforeallowing contactandeventually,feeding.However,itisimportanttonotethat differencesbetweengroupswereonlyamatterofsecondsandare thereforenotlikelytobeclinicallyrelevant.Furtheranalysisofthisphenomenonmayprovideusefulinformationtoresearchersandpro- ducersastohowmotheringupcanbeaffectedbyvarioushusbandry procedures.InTrial2,unrecordedobservationssuggestedthatpain-relatedbehav-iourmayhaveinterferedwithfeed-seekingbehaviourinsomelambs undergoingmulesingwithouttopicalanaesthesia,butthiswasnot apparentincontrollambsorlambstreatedwithtopicalanaesthesia. Againitshouldbenotedthatanydifferencesbetweengroups observedwereonlyamatterof10–20s,whichisunlikelytobeclini-

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ambsorlambstreatedwithtopicalanaesthesia. Againitshouldbenotedthatanydifferencesbetweengroups observedwereonlyamatterof10–20s,whichisunlikelytobeclini- callyrelevant.However,ewesinthesetrialswereheldinarelativelysmallpenof6¥4m,suchthatlambsonlyhadtocrossashortdistancetofindtheirdams.Itispossiblethattheseresultscouldbemagnified inalargerfield.Ourresultsdemonstratingsignificantwoundanaesthesiawithin1minoftheapplicationoftopicalanaesthesiaconcurwithprevious findingsthathavenotedthatmulesingwoundstreatedwithimmedi- atepostoperativetopicalanaesthesiaaredesensitisedwithin1–3min ofmulesing.Assuch,itwouldseempainisunlikelytohaveimpaired thetimestomotheruportofeedinlambstreatedwithtopical anaesthesia.13Theresultsofwoundsensitivitytestingandbehaviouralobservations inthesetrialsareconsistentwithandextendourpreviousfindings thattopicalanaesthesiaformulesingwoundsisefficaciousupto8h post-treatment.13,15Thepresentstudyhasconfirmedanecdotalobser-vationsthatwoundanaesthesiapersistsupto24hpost-mulesing.A significantandincreasinghyperalgesicwoundresponsewasobserved inmulesed,non-anaesthetisedlambsover24hinbothtrialsand thiseffectwassignificantlyamelioratedbytreatmentwithtopical anaesthesia.OurresultsdonotconcursoreadilywithpreviousfindingsbyPaulletal,15whoreportedthatpain-relatedbehavioursinlambstreatedwithtopicalanaesthesiadevelopedbetween4and8hpost-mulesing. Ourprevioustrialshaverevealedminimalevidenceofwoundpainor pain-relatedbehaviourat1,4or8hpost-mulesinginlambstreated withTri-Solfen13andwenowreportminimalevidenceofpain-relatedbehaviourat24hpost-mulesing.Thisvariationmaybeexplainedby differencesinmethodology.Videotapeanalysisoflambbehaviourwas usedinthePaulletal.trial,15whichislikelytobemoresensitivetoabnormalbehavioursthantheNRSfieldobservationsusedinour

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y.Videotapeanalysisoflambbehaviourwas usedinthePaulletal.trial,15whichislikelytobemoresensitivetoabnormalbehavioursthantheNRSfieldobservationsusedinour trials.However,itisalsopossiblethatmethodologyusedbyPaull etal.15trialresultedinexaggeratedpain-relatedbehaviouralresponsesforthelambs.Inthattrial,lambswereheldinsmallindoorpensand underwentrepeatedhandlingat30minand6hpost-treatmentfor bloodsampling,whichmayhaveheightenedtheirstressresponsesand alsoexacerbatedbleedingandpainfromthewoundbecauseofthe Figure7. Mean SEMnumericalratingscoresforpain-relatedbehaviourofthreedif- ferenttreatmentgroupsoflambsfollowing mulesingand/ortaildockingcomparedwith controllambsat1h(grey)and24h(black) aftermulesing(Trial1).HI,hotiron;TA, topicalanaesthesia(Tri-Solfen®);NA,noanaesthesia. PRODUCTIONANIMALSPRODUCTIONANIMALS ©2013TheAuthorsAustralianVeterinaryJournal©2013AustralianVeterinaryAssociationAustralianVeterinaryJournalVolume91,No4,April2013165

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repeatedphysicaldisturbance.Ourtrialsweredesignedtomimicthefieldsituationsuchthatlambswerereturnedtotheirdamsand leftundisturbedinapasture-coveredpaddockbetweenbehavioural assessments.Thismayhavereducedpainandpain-relatedbehaviour intheselambs.ItisimportanttonotethatPaulletal.foundthatlambs treatedwithacombinationoftopicalanaestheticandanon-steroidal antiinflammatorydrug(NSAID)didnotdisplayasignificantoccur- renceofpain-relatedbehavioursincomparisonwithunmulesedcon- trolsbetween4and12h.15ThustheuseofNSAIDsshouldbefurtherexploredandconsideredasanoptionforfurtherreducingpostopera- tivepain.Thetechniqueofwoundsensitivitytestingoverthefirst8hfollowingmulesingwithorwithouttreatmentwithtopicalanaesthesiahasbeen previouslydescribed.13Theresultsfromthepresentstudyindicatethattreatmentwasstilleffectiveat24hpost-mulesingusingthesame woundpainassessmenttechniques.HypersensitivitytoLTandpain stimulationevidentwithinthewoundandthesurroundingareasin thefirstminuteaftermulesinginnon-anaesthetisedlambswasfol- lowedbyincreasedallodyniaandprimaryandsecondaryhyperalgesia at24hpost-mulesing.Therewasevidenceofsignificantandpersist- entwoundanaesthesiaat24hpost-mulesinginlambstreatedwith topicalanaesthesia.Localanaestheticagentsactbyinhibitingthegenerationandconduc-tionofionicfluxesrequiredfortheconductionofnerveimpulses responsibleforthesensationofpain.Theseanaestheticsarereadily absorbedthroughmucousmembranesanddamagedskintoreachthe nervefibres.Tri-Solfencontainslidocainehydrochloride(40.6g/L) andbupivacainehydrochloride(4.5g/L)astheactiveanaesthetic agentsandadrenalinetartrate(24.8mg/L)asavasoconstrictor.The half-lifeforlidocaineinhumansisreportedtobe1.5–2handthatof bupivacaineis2.7hinadultsand8.1hinneonates.Whenusedtopi-

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te(24.8mg/L)asavasoconstrictor.The half-lifeforlidocaineinhumansisreportedtobe1.5–2handthatof bupivacaineis2.7hinadultsand8.1hinneonates.Whenusedtopi- cally,adrenalinewouldslowtherateofsystemicabsorptionofthetwo anaestheticagentsandreducewoundhaemorrhage.Thereducedrate ofsystemicabsorptionkeepstheactiveingredientsconcentratedat thesiteandslowsthemetabolismoftheagents,prolongingtheinten- sityanddurationoflocalanaesthesia.Localanaestheticscanproviderapidandprolongedanaesthesiawhenappliedtoopenwounds19–25andcanalsopreventorreducethesubsequentpainescalationresponse.26–28Thevasoconstrictiveprop-ertiesofadrenalineslowthebloodflowtothewound,therebysup- pressingtheinflammatorycascade,resultinginareductionin associatedpaincausedbyaccumulationofinflammatorymediators, includingcytokinesandhistamines.29Thismayprovideanaddi-tionalexplanationfortheprolongedefficacyofthecombination topicalanaesthesia/vasoconstrictorpreparation.Anotherhypothesisforthelong-termefficacyofthetopicalanaes-thesiaisthepresenceofabarriereffectfrominclusionofagelbasein thepreparation.Wepreviouslyfoundthatmildpainalleviationwas observedthroughtheuseofaplaceboagentthatconsistedofthegel basewithoutthelocalanaestheticorvasoconstrictoractives.13Thisisalsoconsistentwiththefindingsofpublishedstudiesonskinincisions andopenwoundsinhumans,wherebarrierdressingshavesignificant benefitsforwoundhealingandpainattenuationbycoatingdamagednerveendingsandprovidingabarrieragainstongoingenvironmentalexposureandtactilestimulation.30–33ConclusionThetopicalanaesthetic,antisepticandhaemostaticproductTri-Solfen®developedforpainalleviationatmulesinghasaprolongeddurationofactionupto24hpost-treatment.Itishypothesisedthatthecombinationoflocalanaesthesia,haemostaticandvasoconstrictor

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viationatmulesinghasaprolongeddurationofactionupto24hpost-treatment.Itishypothesisedthatthecombinationoflocalanaesthesia,haemostaticandvasoconstrictor actionofepinephrinewithinhibitionoftheinflammatorycascadeand thebarriereffectofthegelbaseoftheproductmayexplainthefinding ofprolongedwoundanaesthesiaobservedinthepresentstudy.AcknowledgmentsThefinancialsupportoftheAustralianResearchCouncil,BayerAnimalHealthAustraliaandAnimalEthicsPtyLtdisgratefully acknowledged.TheauthorsthankMrGeoffMooreandstaffat‘Yerilla’ fortheirtechnicalandlogisticalsupport.Wegreatlyappreciatethe statisticaladvicefromDrPeterThomsonfromtheUniversityof Sydney.References1.BaillieBG.Managementpracticesforcontrollingflystrike.In:ProceedingsoftheNationalSymposiumontheSheepBlowflyandFlystrikeinSheep,NewSouth WalesDepartmentofAgriculture,Sydney,25–27June,1979;159–173. 2.GrahamNPH.TheproblemofflystrikeinsheepinAustralia.In:Proceedingsof theNationalSymposiumontheSheepBlowflyandFlystrikeinSheep,NewSouth WalesDepartmentofAgriculture,Sydney,25–27June,1979;1–5. 3.SimpsonI.Managementproceduresforpreventionandcontrolofflystrike.In: Proceedingsofsheepblowflyandflystrikemanagementworkshop,Trangie, 1987. 4.HartRJ.Reviewofbreechstrikeandthemulesoperation.In:Secondnational symposiumonthesheepblowflyandflystrikeinsheep.NSWDeptofAgriculture, Sydney,1983. 5.DunRB,DonnellyFB.EffectivenessoftheMulesoperationatlambmarking. AustJExpAgricAnimHusb1965;5:6–10.6.ChapmanRE,FellLR,ShuttDA.Acomparisonofstressinsurgicallyand non-surgicallymulesedsheep.AustVetJ1994;71:243–247.7.CroninGM,BarnettJL,EdgeMK,HemsworthPH.Identifyinganimalwelfare issuesforsheepinAustralia.WoolTechnolSheepBreeding2002;50:534–540.8.FellLR,ShuttDA.Behaviouralandhormonalresponsestoacutesurgicalstress

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ganimalwelfare issuesforsheepinAustralia.WoolTechnolSheepBreeding2002;50:534–540.8.FellLR,ShuttDA.Behaviouralandhormonalresponsestoacutesurgicalstress insheep.ApplAnimBehavSci1989;22:283–294.9.GrantC.Behaviouralresponsesoflambstocommonpainfulhusbandry procedures.ApplAnimBehavSci2004;87:255–273.10.JamesP.Geneticalternativestomulesingandtaildockinginsheep:areview. AustJExpAgric2006;46:1–18.11.JamesP.Geneticalternativestomulesingandtaildockinginsheep.AustralianWoolInnovationLimited,Sydney,2004. 12.ScobieDR,O’ConnellD,MorrisCA,HickeySM.Apreliminarygeneticanalysis ofbreechandtailtraitswiththeaimofimprovingthewelfareofsheep.AustJAgricRes2007;58:161–167.13.LomaxS,SheilM,WindsorPA.Impactoftopicalanaesthesiaonpainallevia- tionandwoundhealinginlambsaftermulesing.AustVetJ2008;86:159–168.14.NowakR.Neonatalsurvival:contributionsfrombehaviouralstudiesinsheep. ApplAnimBehavSci1996;49:61–72.15.PaullDR,LeeC,ColditzIG,AtkinsonSJ,FisherAD.Theeffectofatopical anaestheticformulation,systemicflunixinandcarprofen,singlyorincombina- tion,oncortisolandbehaviouralresponsesofMerinolambstomulesing.AustVetJ2007;85:98–106.16.GrahamMJ,KentJE,MolonyV.Effectsoffouranalgesictreatmentsonthe behaviouralandcortisolresponsesof3-week-oldlambstotaildocking.VetJ1997;153:87–97. PRODUCTIONANIMALSPRODUCTIONANIMALS ©2013TheAuthorsAustralianVeterinaryJournal©2013AustralianVeterinaryAssociationAustralianVeterinaryJournalVolume91,No4,April2013166

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17.JongmanEC,MorrisJP,BarnettJL,HemsworthPH.EEGchangesin4-week-oldlambsinresponsetocastration,taildockingandmulesing.AustVetJ2000;78:339–343.18.KentJE,MolonyV.Changesinplasmacortisolconcentrationinlambsofthreeagesafterthreemethodsofcastrationandtaildocking.ResVetSci1993;55:246–251. 19.BrofeldtB,CornwellP,DohertyD,BatraK,GuntherR.Topicallidocaineinthe treatmentofpartial-thicknessburns.JBurnCareRehabil1989;10:63–68.20.JellishWS,GamelliRL,FurryPA,McGillVL,FluderEM.Effectoftopicallocalanestheticapplicationtoskinharvestsitesforpainmanagementinburnpatients undergoingskin-graftingprocedures.AnnSurg1999;229:115–120.21.KokinskyE,CassutoJ,SinclairRetal.Topicalwoundanaesthesiainchildren: atemporarypostoperativepainrelief.ActaAnaesthesiolScand1999;43:225–229.22.SinclairR,CassutoJ,HogstromS.Topicalanaesthesiawithlidocaineaerosolinthecontrolofpostoperativepain.Anaesthesiology1988;68:895–901.23.SinclairR,WestlanderG,CassutoJ,HednerT.Postoperativepainreliefbytopicallidocaineinthesurgicalwoundofhysterectomizedpatients.ActaAnaesthesiolScand1996;40:594–598.24.SmithGA,StrausbaughSD,Harbeck-WeberCetal.Comparisonoftopical anestheticswithoutcocainetotetracaine-adrenaline-cocaineandlidocaineinfil- trationduringrepairoflacerations:bupivacaine-norepinephrineisaneffectivenewtopicalanestheticagent.Pediatrics1996;97:301–307.25.StewartA,SimpsonP,RosenbergN.Useoftopicallidocaineinpediatric lacerationrepair:areviewoftopicalanaesthetics.PediatrEmergCare1998;14:419–423.26.DahlJB,BrennumJ,Arendt-NielsenL,JensenTS,KehletH.Theeffectofpre-versuspost-injuryinfiltrationwithlidocaineonthermalandmechanical

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gCare1998;14:419–423.26.DahlJB,BrennumJ,Arendt-NielsenL,JensenTS,KehletH.Theeffectofpre-versuspost-injuryinfiltrationwithlidocaineonthermalandmechanical hyperalgaesiaafterheatinjurytotheskin.Pain1993;53:43–51.27.LamKW,PunTC,NgEH,WongKS.Efficacyofpreemptiveanalgesiaforwoundpainafterlaproscopicoperationsininfertilewomen:arandomised,doubleblind andplacebocontrolstudy.BiolJObstetGynaecol2004;111:340–344.28.PogatzkiEM,VandermeulenEP,BrennanTJ.Effectofplantarlocalanaesthetic injectionondorsalhornneuronactivityandpainbehaviourscausedbyincision.Pain2002;97:151–161.29.DiasMP,NewtonDJ,McLeodGA,KhanF,BelchJJF.Theinhibitoryeffectsof localanaestheticsonthevascularflareresponsestobradykininandsubstance Pinhumanskin.Anaesthesia2008;63:151–155.30.SchurenJ,BeckerA,SibbaldRG.Aliquidfilm-formingacrylateforperi-wound protection:asystematicreviewandmeta-analysis(3MTMCavilonTMno-stingbarrierfilm).IntWoundJ2005;2:230–238.31.CameronJ,HoffmanD,WilsonJ,CherryG.Comparisonoftwoperi-woundskinprotectantsinvenouslegulcers:arandomisedcontrolledtrial.JWoundCare2005;14:233–236.32.NeanderK,HesseF.Woundedgeprotectioninchronicwounds.JWoundCare2003;12:369–371. 33.RolstadB,BorchertK,MagnanS,ScheelN.Acomparisonofanalcohol-basedandasiloxane-basedperi-woundskinprotectant.JWoundCare1994;3:367–368.(Acceptedforpublication9September2012) BOOKREVIEWColoratlasofveterinaryhistology.3rdedn.EditedbyWJBachaandLMBacha.Wiley-Blackwell,WestSussex,2012.356pages.PriceA$125.00.ISBN9780470958513.The3rdeditionofthishardcovertextcovershistologyinthemajorveterinaryspecies(dog,cat,horse,cow,sheep,goat,pigandchicken)andisdesignedasalearningtoolforstudents.Thebookbeginswithabriefintroductiontohistology,includingtipsforviewingslidesandsettingupamicroscope.Alltissuesand

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isdesignedasalearningtoolforstudents.Thebookbeginswithabriefintroductiontohistology,includingtipsforviewingslidesandsettingupamicroscope.Alltissuesand systemsarediscussedchapterbychapter.Thereisashortintroduc-tiontoeachtissueorsystem,followedbyannotatedphotomicro-graphsrelevanttotheappropriatespecies.Allphotomicrographs areincolouranddiagramsareinblackandwhite.Thechapteron haematologyispredominantlyillustratedwithbloodsmearsrather thanhistologicalsections.Itidentifiesthenormalbloodcompo- nentsforeachspeciesandmentionscommonvariationsseenon normalsmears.Thetextconcludeswithaglossaryandindex.Theintroductorychapterisshortandsuccinct,butcoversallthatwouldberequiredbymostveterinarystudents.Headingsarecon- sistentandeasilyrecognisedandthephotomicrographsarewell laidout.Boxescontainingchaptersummariesarenotalwaysclear ataglance,becauseofpoortextformatting.Helpfulhints,high- lightedinblueboxes,areveryusefulandcovertopicssuchashow torecognisecertaintissuesandsuggestionsforexaminingcertain tissuesandslides.Theglossarymaybeusefulforsome,butthe internetislikelytohavemadethissectionsomewhatredundant.Althoughmostoftheillustrationsareofhighquality,aproportionisnot,particularlythelargerpictures.Thechapteronbonemarrowisrudimentaryandwouldnotbeofparticularusetoeitherstudentsorpathologistsasareference.Thiseditionisslightlylongerandthelayoutofthetextimprovedandmoreuserfriendly,especiallytheheadings,comparedwiththe 2ndedition,butthecontentisessentiallyunchanged.Helpfulhints androotwordboxesareausefuladdition.Mostoftheillustrations fromthe2ndeditionareincluded,withsomenewones.TheCDisnewforthe3rdedition,butcontainsonlytheillustrations,which areviewedviaanindexorsearchfunction.Thetextisillustrativeratherthananin-depthdescriptionoftissue.Manyofthemedicalhistologybookshavesuchdetailedanalysis

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reviewedviaanindexorsearchfunction.Thetextisillustrativeratherthananin-depthdescriptionoftissue.Manyofthemedicalhistologybookshavesuchdetailedanalysis andbrieflycoverEMandcellcycles,topicsthatarenotcoveredin thisatlas.However,Ilovetheextensivecoverageofspecies-specific variations,whichobviouslycannotbefoundinamedicalhistology text.Thetextwouldbehelpfultostudentsstudyingveterinaryhistologyandasareferenceforbothveterinariansinpracticeandveterinary pathologists.PriscillaHodgedoi:10.1111/avj.12035 PRODUCTIONANIMALSPRODUCTIONANIMALS ©2013TheAuthorsAustralianVeterinaryJournal©2013AustralianVeterinaryAssociationAustralianVeterinaryJournalVolume91,No4,April2013167