Vrije Universiteit Brussel

Courtship pheromones in mudpuppies and waterdogs (Necturus, Proteidae)

Treer, Dag; Maex, Margo; Stoops, Monica A.; Nagel, Andrew; DeChant, Chris J.; VanBocxlaer, Ines; Bossuyt, Franky

Publication date:2017

Document Version:Final published version

Link to publication

Citation for published version (APA):Treer, D., Maex, M., Stoops, M. A., Nagel, A., DeChant, C. J., Van Bocxlaer, I., & Bossuyt, F. (2017). Courtshippheromones in mudpuppies and waterdogs (Necturus, Proteidae). 29-29. Poster session presented at 14thMeeting of Chemical Signal in Vertebrates (CSiV XIV), Cardiff, United Kingdom.

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Download date: 24. Aug. 2020

14thMeetingoftheCSiVgroup,SchoolofBiosciences,CardiffUniversity,UK,

29August–1September2017

ProgrammeandAbstractBooklet

29August– 1September2017Cardiff,Wales,UK

2

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TableofContents

Programme........................................................................................................................6

AbstractsofTalksandPostersinorderofAppearance.....................................................13

Tuesday29August2017...................................................................................................13

ThematicSessionI:HumanChemicalCommunication......................................................13Talk:Theeffectofdietaryrestrictionsonhumanbodyodourquality........................................13Talk:DriversofMHC-dependentmatechoiceinhumansandprimates......................................13Talk:Canyousmellthewinner?Theeffectofcompetitiononhumanbodyodour....................14Talk:Evidenceandthoughtsabouttheinfluenceofsex,genderandcultureonhumanbodyodourperception.......................................................................................................................14Talk:Olfactorily-conspicuousnipplesasvitalinterfacesforcolostrumintakeinhumans...........15Talk:Perfumepreferencesinhumancouples.............................................................................16Talk:Investigatingtheartificiallyscentedape:Theimpactofartificialfragrancesonhumanolfactorycommunication...........................................................................................................16Talk:Chemocommunicationinavision-ruledworld...................................................................17Talk:Sexinguphumanpheromones:Howacorporationcreatedamythandleftatrailoffalsepositivespavedbypositivepublicationbias..............................................................................18

Opensession1.................................................................................................................18Talk:PheromoneofIndianLeopard,Pantherapardus...............................................................18Talk:Infantpandascannotdiscriminatebetweenteatodorsofdifferentlactatingfemales......19Talk:DecodingMouseSocialCommunication............................................................................20

OpenSession2.................................................................................................................20Talk:Whatmayattractbluepetrelstotheirburrowsatnight?..................................................20Talk:HarderianGlandConstituentsMediateVomeronasalFunctioninginGarterSnakes..........21Talk:Effectsoflowenvironmentalcalciumonresponsestofoodandpredator-relatedodoursinfatheadminnows.......................................................................................................................22

Wednesday30August2017.............................................................................................22

PLENARY:Underwaterchemistrybetweensalamanders:Evolutionofacourtshippheromonesystem..........................................................................................................22

ThematicSessionII:Proteomics.......................................................................................23Talk:ChallengesinproteomicsoftheMUPmulti-genefamily....................................................23Talk:Chemosignalsinanuranbreedingglands...........................................................................23Talk:Anewcandidatecourtshippheromoneinaquaticallycourtingnewts...............................24Talk:MolecularcomplexityofscentsignalsintheNorwayrat..................................................25Talk:Impactofmaleeffectandanoestrusoestrusstatusesongoatolfactorysecretome...........25Talk:Identificationandcharacterizationofaseasonally-expressedsexuallydimorphicurinarylipocalininthebankvoleMyodesglareolus...............................................................................26Talk:Apreliminarystudyontheanalsacproteomeindogs(Canisfamiliaris)............................27

PostertalksI:Proteomics................................................................................................27

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POSTER1:Beyondsodefrin:evidenceforamulti-componentpheromonesysteminthemodelnewtCynopspyrrhogaster(Salamandridae)..............................................................................27POSTER2:Seasonalchangesinolfactoryproteinsofewe..........................................................28POSTER3:Courtshippheromonesinmudpuppiesandwaterdogs(Necturus,Proteidae)...........29POSTER4:Anon-conventionalglycosylationisinvolvedinthecodingofspecificodorsindomesticpig..............................................................................................................................29

ThematicSessionIII:OlfactorySpeciation........................................................................30Talk:Comparativeproteinstructurepredictionandelucidationofbovinevomeronasal1receptortype-I(VN1R1)adoptingin-silicostudy......................................................................30Talk:Anovellipidmetabolismsystemdevelopedincatsproducesaspecies-specificchemosignal..................................................................................................................................................31Talk:OlfactoryspeciationinEurasianbadgersMelesspp.:Diversityofsubcaudalchemicalsandbehaviouralexperiments...........................................................................................................31Talk:Odourdialectsamongwildmammals................................................................................32

Postertalks2:Proteomics&Primateolfaction.................................................................32POSTER5:QuantitativeProfilingofMajorUrinaryProteinsinWildMice...................................32POSTER6:Identificationandcharacterizationofaseasonally-expressedsexuallydimorphicurinaryproteinintwospeciesofMalagasymouselemurs,MicrocebusmurinusandMicrocebuslehilahytsara.............................................................................................................................33POSTER7:Makingsenseofscents:Decipheringprimateolfactorycommunication....................34POSTER8:EffectsofMaleAxillaryExtractsontheLengthofMenstrualCycleinReproductiveAgeWomenandPre-MenopausalWomen.......................................................................................34

OpenSession3.................................................................................................................35Talk:Socialodorspromotefood-relatedpreferencelearningthroughactivationoftheGC-D/necklaceolfactorysubsystem................................................................................................35Talk:Influencesofexposuretomurinepheromoneonthenextgeneration...............................36

Thursday31August2017.................................................................................................36

PLENARY:Domammalshaveaworldwidewebofinterspecificscentsignals,andcouldweconnecttoit?...................................................................................................................36

ThematicSessionIV:Inter-specificolfactorycues(Chair:PeterApps)...............................37Talk:EarlyOlfactoryExperienceAffectsPerceptionofPredatorOdorsintheHouseMouse......37Talk:Ontherepellentpropertiesofmouseurineodourtosmallwildlivingrodents..................37

PosterTalks3:Inter-specificCues&OtherContributions.................................................38POSTER9:EffectofDomesticCatChemicalSignalsonRegulationofEstrousCyclesintheHouseMouseDependsonPresentationRate.......................................................................................38POSTER10:Preliminaryresultsinferretolfactorycues(Mustelaputoriusfuro)asapredatorstimulusforthehousemice(Musmusculus)..............................................................................39POSTER11:Measuringlactationinvestmentamongcommunallynursingfemalehousemice....39POSTER12:Latrineusepatternsofraccoondogs(Nyctereutesprocyonoides)inanurbanareaofTokyo........................................................................................................................................40Poster13:Tastepotentiatedcolourbiases:analternativemethodforassessingtastedetectionthresholdsinbirds.....................................................................................................................41

5

ThematicSessionV:ScentofDisease...............................................................................41Talk:LPS-inducedimmunesystemstimulationalterurinaryvolatilesandbehavioringrowingpigs............................................................................................................................................41Talk:Sharinganenvironmentwithsickconspecificsalterstheodorsofhealthymice................42Talk:Diseasedetectioninhumanolfaction................................................................................42Talk:Thesmellofweakness:costsandconsequencesofinjuryforsocialodorcuesinring-tailedlemurs.......................................................................................................................................43

ThematicSessionVI:Semio-chemicalMethodologiesandAnalyses..................................44Talk:ComplexityofScentMarkinginRats:BehaviouralCharacterisation..................................44Talk:PTR-TOF-MSasatoolforstudyingvertebratechemicalecology:acasestudyontheuseinkairomonedeterminationinapredator-preycontext................................................................44Talk:Non-invasivesamplingofbodyodourtoinvestigatetheinformationcontentofmeerkatscent..........................................................................................................................................45Talk:Themaleeffect:findingawaythroughthejungleofchemicalcues..................................46Talk:Comparisonofdifferentanalyticalmethodsforpre-processinggaschromatographicdatainsemio-chemicalresearch........................................................................................................46Talk:SamplingandanalysisofVOCs&SVOCs–Whatarethebesttoolsforthejob?..................47

Opensession4.................................................................................................................48Talk:Temporalchemicalprofilingofurinevolatilecompoundsofdomesticcatsandtheirolfactorydiscriminationabilities................................................................................................48Talk:Identificationofkeycompoundsfortheindividualrecognitioninthedomesticcats.........48

Friday01September2017................................................................................................49

Plenary:Scentwars:unravellingthecomplexityofcompetitivescentsignalling...............49

ThematicSessionVII:Olfactoryadvertisement.................................................................50Talk:Urinaryvolatilesandbioindicatorsrelatedtoreproductivestatusingiantpanda(Ailuropodamelanoleuca).........................................................................................................50Talk:Femalesprefercooperativemalesbasedoncuesdifferentfromcooperativebehaviours:evidencefromtheMound-buildingmouse,Musspicilegus........................................................50Talk:Sex-steroidexcretioninurineasolfactoryadvertisementinEuropeanbadgers(Melesmeles)inrelationtocirculatingplasmatitres............................................................................51Talk:Scent-markingasHonestFitnessAdvertisementinEuropeanBadgers(Melesmeles)........51

PosterTalks4:Advertisement..........................................................................................52POSTER14:Changesincompositionofurineduringvariousperiodsofovariancycleinfemaleofdomesticdogs(Canisfamiliaris)................................................................................................52POSTER15:ScentCommunicationinBankVoles:Doesmaleinvestmentinfluencefemaleresponse?..................................................................................................................................53POSTER16:ExploringVOCsfromharbourporpoiseexcreta.......................................................53POSTER17:Putativepheromoneofsnowleopard,Pantherauncia(Schreber,1775).................54

Listofdelegates:..............................................................................................................55

6

Programme

Monday28.08.2017

17.30-21.00:RegistrationandWelcomeReception

Tuesday29.08.2017

8.30-9.15:WelcomeandOpeningSessionfortheCSiVXIV:CarstenTMüller&ChristinaDBuesching

9:20-13:00ThematicSessionI:Humanchemicalcommunication(chair:BenoistSchaal,JanHavlicek,&CraigS.Roberts)9.20-9.30:Introduction

9.30-9.50:Theeffectofdietaryrestrictionsonhumanbodyodourquality(J.Havlíček,J.Fialová&S.C.Roberts)

9.50-10.10:DriversofMHC-dependentmatechoiceinhumansandprimates(J.L.Abbate,J.Winternitz,E.Huchard,J.Havlíček&L.Z.Garamszegi)

10.10-10.30:Canyousmellthewinner?Theeffectofcompetitiononhumanbodyodour(JFialová,VTřebický&JHavlíček)

10.30-10.50:Evidenceandthoughtsabouttheinfluenceofsex,genderandcultureonhumanbodyodourperception(C.Ferdenzi,A.FournelArnaud&M.Bensafi)

10.50-11.20:Coffee

11.20-11.40:Olfactorily-conspicuousnipplesasvitalinterfacesforcolostrumintakeinhumans(BSchaal,M-LSeydel,SDoucet&KDurand)

11.40-12.00:Perfumepreferencesinhumancouples(S.C.Roberts,E.Simpson,J.Havlicek&C.Allen)

12.00-12.20:Investigatingtheartificiallyscentedape:Theimpactofartificialfragrancesonhumanolfactorycommunication(C.Allen,J.Havlíček,K.D.Cobey&S.C.Roberts)

12.20-12.40:Chemocommunicationinavision-ruledworld(J-YBaudouin,KDurand,A.Leleu,ASeigneuric&BSchaal)

7

12.40-13.00:Sexinguphumanpheromones:Howacorporationcreatedamythandleftatrailoffalsepositivespavedbypositivepublicationbias(T.Wyatt)

13.00-14.00:Lunch

14:00-15:20Opensession1(Chair:ChristinaDBuesching)

14.00-14.20:PheromoneofIndianLeopard,Pantherapardus(B.Ghosh,S.Das,P.Das,S.Manna,C.S.Pal,S.M.Poddar&R.L.Bhamachary)

14.20-14.40:Infantpandascannotdiscriminatebetweenteatodorsofdifferentlactatingfemales(D.Liu,X.Xie,H.Wu,B.Luo&G.Zhang)

14.40-15.00:DecodingMouseSocialCommunication(X.Fu,Y.Yan,P.Xu,IGeerling-Vidavsky,M.Gross&T.Holy)

15.00-15.50:COFFEE

15:50-16:50OpenSession2(CarstenTMüller)

15.50-16.10:Whatmayattractbluepetrelstotheirburrowsatnight?(M.Gabirot,F.Bonadonna&C.T.Müller)

16.10-16.30:HarderianGlandConstituentsMediateVomeronasalFunctioninginGarterSnakes(E.J.Bentz&R.T.Mason)

16.30-16.50:Effectsoflowenvironmentalcalciumonresponsestofoodandpredator-relatedodoursinfatheadminnows(R.S.Mirza&C.Cornell)

PubCrawl(leaving19:30h)

8

Wednesday30.08.2017

8.30-9.30:Plenary:FrankyBossuyt:Underwaterchemistrybetweensalamanders:Evolutionofacourtshippheromonesystem

9:30-12:40ThematicSessionII:Proteomics(Chair:SunitaJanssenswillen)

9.30-9.50:ChallengesinproteomicsoftheMUPmulti-genefamily(R.J.Beynon,A.J.Davidson,G.Gomez-Baena,V.M.Harman,L.McLean,M.Prescott,S.A.Roberts&J.L.Hurst)

9.50-10.10:Chemosignalsinanuranbreedingglands(L.M.Schulte,S.Janssenswillen,K.Roelants,S.Matthijs,P.Kok,I.VanBocxlaer&F.Bossuyt)

10.10-10.30:Anewcandidatecourtshippheromoneinaquaticallycourtingnewt(M.Maex,D.Treer,S.Janssenswillen,I.VanBocxlaer&F.Bossuyt)

10.30-10.50:MolecularcomplexityofscentsignalsintheNorwayrat(G.Gómez-Baena,K.Pounder,J.O.Halstead,S.D.Armstrong,M.Prescott,S.A.Roberts,L.McLean,J.L.Hurst,&R.J.Beynon)

10.50-11.20:Coffee

11.20-11.40:Impactofmaleeffectandanoestrusoestrusstatusesongoatolfactorysecretome(M.Chabi,C.LeDanvic,M.Chasles,M.Keller,P.Nagnan-LeMeillourP)

11.40-12.00:Identificationandcharacterizationofaseasonally-expressedsexuallydimorphicurinarylipocalininthebankvoleMyodesglareolus(G.M.Loxley,J.Unsworth,M.Turton,A.Davidson,A.Jebb,J.L.Hurst&R.J.Beynon)

12.00-12.20:Apreliminarystudyontheanalsacproteomeindogs(Canisfamiliaris)(S.Janssenswillen,M.Metzemaekers,P.Proost&F.Bossuyt)

12.20-12.40:PostertalksI:Proteomics

POSTER1:Beyondsodefrin:evidenceforamulti-componentpheromonesysteminthemodelnewtCynopspyrrhogaster(Salamandridae)(I.VanBocxlaer,M.Maex,D.Treer,S.Janssenswillen,R.Janssens,W.Vandebergh,P.Proost&F.Bossuyt)

POSTER2:Seasonalchangesinolfactoryproteinsofewe(P.Cann,M.Chabi,C.LeDanvic,R.Lami,M.Keller&P.Nagnan-LeMeillour)

POSTER3:Courtshippheromonesinmudpuppiesandwaterdogs(Necturus,Proteidae)(D.Treer,M.Maex,M.A.Stoops,A.Nagel,C.J.DeChant,I.VanBocxlaer&F.Bossuyt)

POSTER4:Anon-conventionalglycosylationisinvolvedinthecodingofspecificodorsindomesticpig(A.Joly,S.Potelle,C.Spriet,C.LeDanvic,P.NagnanLeMeillour)

12.20-14.00:Lunch

9

ThematicSessionIII:OlfactorySpeciation

14.00-14.20:Comparativeproteinstructurepredictionandelucidationofbovinevomeronasal1receptortype-I(VN1R1)adoptingIn-silicostudy(DurairajRajesh,C.Bienboire-Frosini&P.Pageat)

14.20-14.40:Novellipidmetabolismsystemdevelopedincatsproducesaspecies-specificchemosignal(M.Miyazaki,A.Futsuta,W.Hojo,T.Miyazaki&T.Yamashita)

14.40-15.00:OlfactoryspeciationinEurasianbadgersMelesspp.:Diversityofsubcaudalchemicalsandbehaviouralexperiments(Y.Kaneko,C.Newman,E.Raichev,S.Kosuga,S.Yachimori,M.Saito,Y.Kuramoto,D.W.Macdonald,S.Peeva&C.D.Buesching)

15.00-15.20:Odourdialectsamongwildmammals(E.F.Kean,M.W.Bruford,I.M.Russo,C.T.Müller&E.A.Chadwick)

15.20-16.10:COFFEE

16.10-16.30:Postertalks2:Proteomics(cont.)&Primateolfaction

POSTER5:QuantitativeProfilingofMajorUrinaryProteinsinWildMice(M.C.Prescott,G.Gómez-Baena,V.M.Harman,A.J.Davidson,S.A.Roberts,J.L.Hurst&R.J.Beynon)

POSTER6:Identificationandcharacterizationofaseasonally-expressedsexuallydimorphicurinaryproteinintwospeciesofMalagasymouselemurs,MicrocebusmurinusandMicrocebuslehilahytsara(G.Loxley,A.Trester,J.Unsworth,G.Gomez-Baena,U.Radespiel,E.Zimmermann,J.L.Hurst,R.J.Beynon)

POSTER7:Makingsenseofscents:Decipheringprimateolfactorycommunication(A.Poirier)

POSTER8:EffectsofMaleAxillaryExtractsontheLengthofMenstrualCycleinReproductiveAgeWomenandPre-MenopausalWomen(V.V.Voznessenskaya&T.K.Laktionova)

16:30-17:10OpenSession3

16.30-16.50:Socialodorspromotefood-relatedpreferencelearningthroughactivationoftheGC-D/necklaceolfactorysubsystem(A.D.Zimmerman,H.Zhi&S.D.Munger)

16.50-17.10:Influencesofexposuretomurinepheromoneonthenextgeneration(S.Koyama,H.Soini,U.Tadasu&M.Novotny)

10

Thursday31.08.2017

8.30-9.30: Plenary: Peter Apps: Do mammals have a world wide web ofinterspecificscentsignals,andcouldweconnecttoit?

9:30-10:10ThematicSessionIV:Inter-specificolfactorycues(Chair:P.Apps)

9.30-9.50:EarlyOlfactoryExperienceAffectsPerceptionofPredatorOdorsintheHouseMouse(V.V.Voznessenskaya,I.G.Kvasha,T.K.Laktionova,A.B.Klinov&M.A.Klyuchnikova)

9.50-10.10:Ontherepellentpropertiesofmouseurineodourtosmallwild-livingrodents(E.V.Kotenkova,I.A.Zhigarev,V.V.Alpatov,A.O.Shchukin,V.A.Babikov&Y.A.Bazhenov)

10.10-10.40:PosterTalks3:Inter-specificcues&Othercontributions

POSTER9:EffectofDomesticCatChemicalSignalsonRegulationofEstrousCyclesintheHouseMouseDependsonPresentationRate(T.K.Laktionova,I.Kvasha&V.V.Voznessenskaya)

POSTER10:Preliminaryresultsinferretolfactorycues(Mustelaputoriusfuro)asapredatorstimulusforthehousemice(Musmusculus)(P.C.Grau,J.Leclerq,E.Teruel,C.Lafont-Lecuelle&P.Pageat)

POSTER11:Measuringlactationinvestmentamongcommunallynursingfemalehousemice(A.Davidson,J.P.Green,C.DeMatosFerrazFranco,V.Lee,PStockley,R.J.Beynon&J.L.Hurst)

POSTER12:Latrineusepatternsofraccoondogs(Nyctereutesprocyonoides)inanurbanareaofTokyo(M.Tsunoda,Y.Kaneko,T.Sako,R.Koizumi,K.Iwasaki,I.Mitsuhashi,M.U.Saito,C.Newman,D.W.Macdonald&C.D.Buesching)

POSTER13:Tastepotentiatedcolourbiases:analternativemethodforassessingtastedetectionthresholdsinbirds(H.M.Rowland)

10.40-11.20:COFFEE

11:20-13:00ThematicSessionV:ScentofDisease(Chair:StefanieGervasi)

11.20-11.40LPS-inducedimmunesystemstimulationalterurinaryvolatilesandbehavioringrowingpigs(S.Devaraj,A.Rakhshandeh&J.McGlone)

11.40-12.00:Sharinganenvironmentwithsickconspecificsalterstheodorsofhealthymice(S.Gervasi,B.A.Kimball,M.Opiekun,T.Martin&G.K.Beauchamp)

12.00-12.20:Diseasedetectioninhumanolfaction(M.Olsson)

11

12.20-12.40:Thesmellofweakness:costsandconsequencesofinjuryforsocialodorcuesinring-tailedlemurs(R.Harris&C.Drea)

12.40-14.00:LUNCH

14:00-15:40ThematicSessionVI:Semio-chemicalMethodologiesandAnalyses(ChairCDBuesching)

14.00-14.20:ComplexityofScentMarkinginRats:BehaviouralCharacterisation(K.C.Pounder,G.Gómez-Baena,J.O.Halstead,M.Prescott,R.J.Beynon&J.L.Hurst)

14.20-14.40:PTR-TOF-MSasatoolforstudyingvertebratechemicalecology:acasestudyontheuseinkairomonedeterminationinapredator-preycontext(C.VanMoorleghem,M.Portillo-Estrada,Y.Lambreghts,I.A.Janssens&R.VanDamme)

14.40-15.00:Non-invasivesamplingofbodyodourtoinvestigatetheinformationcontentofmeerkatscent(B.M.Weiß,A.Marcillo,M.Kücklich,M.Manser,R.Holland,C.Birkemeyer&A.Widdig)

15.00-15.20:Themaleeffect:findingawaythroughthejungleofchemicalcues(C.LeDanvic,M.Dillenbourg,M.Keller,L.Schibler&P.Nagnan-LeMeillour)

15.20-15.40:Comparingdifferentanalyticalpre-processingmethodsofGCMSdatainecologicalresearch(C.D.Buesching,H.V.Tinnesand,C.T.Mueller&M.J.Noonan)

15.40-16.00:SamplingandanalysisofVOCs&SVOCs–Whatarethebesttoolsforthejob?(MassimoSantoro,IlariaFerrante,HannahCalder)

16:00-16.40:COFFEE

16:10-15:50Opensession4

16.30-16.50:Temporalchemicalprofilingofurinevolatilecompoundsofdomesticcatsandtheirolfactorydiscriminationabilities(ChiharuSuzuki,MinoruMaita,TamakoMiyazaki,TetsuroYamashita&MasaoMiyazaki)

16.50-17:10:Identificationofkeycompoundsfortheindividualrecognitioninthedomesticcats(MinoruMaita,ChiharuSuzuki,JanaCaspers,TetsuroYamashita,StefanSchulz&MasaoMiyazaki)

19.30-10.00:ConferenceDinner

12

Friday01.09.2017

8.30-9.30: Plenary: Jane Hurst: Scent wars: unravelling the complexity ofcompetitivescentsignalling

9:30-10:50ThematicSessionVII:Olfactoryadvertisement(ChairJaneHurst)

9.30-9.50:Urinaryvolatilesandbioindicatorsrelatedtothereproductivestatusingiantpanda(Ailuropodamelanoleuca)(M.Dehnhard&J.Brown)

9.50-10.10:Femalesprefercooperativemalesbasedoncuesdifferentfromcooperativebehaviours:evidencefromtheMound-buildingmouse,Musspicilegus(A.Tognetti,M.Raymond,G.Ganem&C.Faurie)

10.10-10.30:Sex-steroidexcretioninurineasolfactoryadvertisementinEuropeanbadgers(Melesmeles)inrelationtocirculatingplasmatitres(N.A.Sugianto,H.V.Tinnesand,D.W.Macdonald&C.D.Buesching)

10.30-10.50:Scent-markingasHonestFitnessAdvertisementinEuropeanBadgers(Melesmeles)(T.Allen,C.D.Buesching&D.W.Macdonald)

10.50-11.30:Coffee

11.30-11.50:PosterTalks4:Advertisement

POSTER14:Changesincompositionofurineduringvariousperiodsofovariancycleinfemaleofdomesticdogs(Canisfamiliaris)(M.Dzięcioł,A.Szumny,W.Niżański,T.Jezierski,M.Woszczyło&B.Pieczewska)

POSTER15:ScentCommunicationinBankVoles:Doesmaleinvestmentinfluencefemaleresponse?(H.Coombes,Stockley,R.J.Beynon&J.L.Hurst)

POSTER16:ExploringVOCsfromharbourporpoiseexcreta(M.Gabirot,B.Bouchard,A.Célérier,L.IJsseldijk&S.Campagna)

POSTER17:Putativepheromoneofsnowleopard,Pantherauncia(Schreber,1775)(S.Das,P.L.Das&M.P.Sarkar)

12.00-13.00:ClosingsessionandplanningofnextCSiVconference

13.00-14.00:Lunch

14.30:Excursions(CardiffCastle,DrWhoExperience)

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AbstractsofTalksandPostersinorderofAppearance

Tuesday29August2017

ThematicSessionI:HumanChemicalCommunication(ChairB.Schaal,J.Havlicek&C.S.Roberts)

Talk:TheeffectofdietaryrestrictionsonhumanbodyodourqualityJ.Havlíček1,2,J.Fialová1,2&S.C.Roberts3

1FacultyofScience,CharlesUniversity,Prague,CzechRepublic;2NationalInstituteofMentalHealth,Klecany,CzechRepublic;3UniversityofStirling,Stirling,UKPreviousstudiesonvariousvertebrateshaveshownthatquantityandqualityoffoodintakeaffectodourattractivenessasperceivedbypotentialmates.Similarly,inhumans,thequalityofbodyodourisaffectedbyingestedfoods,suchasmeatandgarlicintake.Nevertheless,itisnotknownwhetherquantityoffoodhasanimpactonbodyodourattractiveness.Thus,herewetestedhowstarvationaffectsthehedonicqualityofhumanaxillaryodour.Odoursamples(cottonpadsfixedinbotharmpitsandwornfor12hours)wereobtainedfromhealthyfemaledonorsacrossthreeconditions:i)duringtheirhabitualfoodregime;ii)after48hoursofstarvation(drinkingwaterwasprovided),andiii)72hoursafterrestorationofcaloricintake.Axillarysampleswereassessedbymaleratersregardingtheirpleasantness,attractivenessandintensity.Wealsocollectedbloodsamplesforglucoselevelassays.Theodoursamplescollectedduringstarvationwereratedsignificantlylesspleasant,lessattractiveandmoreintensecomparedtoodoursamplescollectedafterrestorationoffeeding.Interestingly,wealsofoundthatbodyodourwasratedmorepleasant,moreattractiveandlessintenseafterrestorationoffoodintakethanpriortostarvation.Wealsocalculateddifferencesinratingsofsamplescollectedduringstarvingandafterstarving.Theincreaseinodourattractivenessasfeedingresumedwasnegativelyrelatedtoglucosechangeduringthesameperiod.Ourresultssuggestthatrestorationoffoodintakepositivelyinfluenceshedonicqualityofhumanbodyodourwhichmightthusprovidecuestocurrentfitnessstatusandmetabolicefficiency.

Talk:DriversofMHC-dependentmatechoiceinhumansandprimatesJ.L.Abbate1,J.Winternitz2,E.Huchard3,J.Havlíček4&L.Z.Garamszegi5

1CBGPInra(UMR1062Inra-IRD-Cirad-MontpellierSupAgro),Montferrier-sur-Lez,France2DepartmentofEvolutionaryEcology,MaxPlanckInstituteforEvolutionaryBiology,Ploen,Germany3CEFECNRS(UMR5175CNRS-UniversitédeMontpellier-EPHE),Montpellier,Fance4DepartmentofZoology,FacultyofScience,CharlesUniversity,Prague,CzechRepublic5DepartmentofEvolutionaryEcology,EstaciónBiológicadeDoñana-CSIC,41092,Seville,Spain

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Genesofthemajorhistocompatibilitycomplex(MHC)invertebratesareintegralforeffectiveadaptiveimmuneresponseandareassociatedwithsexualselection.EvidencesupportsMHC-basedpreferencefordiverseanddissimilarmatingpartners,inarangeofvertebratesfromfishtononhumanprimates.However,evidencefromhumanmatechoicestudieshasbeenmorecontroversial.Here,weperformedaphylogeneticallycontrolledmeta-analysiswithmeta-regressiontechniquesusing58effectsizesfrom30studiesacross7primatespeciesinanefforttounderstandwhatprocessesmayimpactMHC-mediatedmatechoiceinhumansandnon-humanprimates.Bothhumanandnon-humanprimatestudiesshowedevidenceofmateselectionforMHC-diversity.Incontrast,therewasnogeneraltrendforMHC-dissimilarity,andforhumans,effectsizeswereobservedforbothMHC-dissimilarandsimilarmates.Couplesprimarilyshowedsimilaritypreferences,suggestingtheinfluenceofsocio-culturalassortativebiases.

Talk:Canyousmellthewinner?TheeffectofcompetitiononhumanbodyodourJ.Fialová1,2,V.Třebický1,2&J.Havlíček1,2

1FacultyofScience,CharlesUniversity,Prague,CzechRepublic;2NationalInstituteofMentalHealth,Klecany,CzechRepublic;Previousresearchhassuggestedthatsomevertebratessuchasrodentsuseodourcuestoassessindividuals´competitiveabilitiesordominance.Similarly,itwasshownthattheodourofdominantmalesisperceivedbywomenasmoreattractiveandbodyodourcollectedduringcompetitionelicitshigherskinconductanceresponsecomparedtonon-competitivecontext.Here,weinvestigatedwhetherwinningandlosingamatch,amarkerofpossiblechangeindominancehierarchy,willinfluencehedonicvalenceofhumanbodyodour.Weemployedmixedmartialarts(MMA)asamodelofreal-lifephysicalencounters.Wecollectedbodyodoursamples(samplingoncottonT-shirtsfor30minutes)fromasetofMMAfightersbeforeandaftertheirfightonCzechAmateurMMAleague.Obtainedstimuliweresubsequentlyratedapanelofratersregardingtheirpleasantness,attractiveness,andintensity.Wefoundsignificantdifferencesinbodyodoursamplescollectedbeforeandafterthefightinindividualswholosttheirfight.Theirbodyodourwasratedaslessattractiveafterthefight.Incontrast,therewasnochangeinodourattractivenessinthewinners.Further,nosimilarpatternwasobservedinpleasantnessandintensity.Ourresultsshowsignificanteffectofcompetitionoutcomeonperceivedqualityofhumanbodyodourwithdecreasedattractivenessinlosers.Theseresultsmightbeduetothedifferencesinaffectivestateswhichwerepreviouslyshowntoaffectthequalityofbodyodour.

Talk:Evidenceandthoughtsabouttheinfluenceofsex,genderandcultureonhumanbodyodourperception

C.Ferdenzi,A.Fournel&M.Bensafi

LyonNeuroscienceResearchCenter,CNRS-Inserm-UniversityClaudeBernardLyon1,Lyon,France.

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Althoughsocialcommunicationisoneofthemajorfunctionsofolfactioninnumerousspecies,inhumansmanyaspectsofthisfunctionremainpoorlyunderstood.Thereisincreasingevidencethatodoursnaturallyemittedbythehumanbodyareextremelyimportantininterpersonalrelationshipsandthattheyinfluencemateattractiveness.However,ourknowledgeisonlypartialwhenitcomestothenatureofthechemicalcompoundsinvolvedbutalsotothesocio-culturalmodulationofhumanolfactorysocialcommunication.Whilemanystudieshavefocusedonandrostenesaspossiblyrelevantchemosignals,othercompoundssuchasthoseproducedinsex-specificamountsdeserveattention.Here,Iwillpresentseveralstudiesinvestigatinghowsomeofthelattercompoundsareperceivedandinfluencepersonperception,differentiallyinmenandwomen.Iwillalsodiscusschallengesforfutureresearchinthisarea,suchastheunderstandingtheoriginofsuchsexdifferences,andespeciallyhowsocio-culturalfactorsmayshapetheminaccordancewithsocialnorms,individual’sexperienceandcurrentneeds.Anotherstimulatingchallengeistousecross-culturalresearchtoincreaseourunderstandingofhowbodyodoursinfluencehumansocialinteractions.Todevelopthisidea,Iwillpresentasetofcross-culturalstudiesaboutodour-relatedfeelingsandattention,withaparticularfocusonaspectsrelatedwithpeople’sodour.Thiswilloffersomethoughtsonpossiblefuturedirections,combiningthestudyofculturaldifferencesinhumanolfactionandthestudyofsocialcommunicationthroughbodyodours.

AMCoffeebreak

Talk:Olfactorily-conspicuousnipplesasvitalinterfacesforcolostrumintakeinhumansB.Schaal,M.-L.Seydel,S.Doucet&K.Durand

DevelopmentalEthologyandCognitivePsychologyGroup,CenterforSmell,TasteandFoodScience,CNRS-UniversitédeBourgogne,Dijon,France

Afterbirth,fragilenewbornsmustingestthecolostrumasswiftlyaspossibletoincorporaterelatedbenefitssuchasimmuno-protection,growthofdigestivetract,andcolonizationbygutmicrobiota.Reciprocally,femalesmustrespondtimelytotheiroffspringbecauseanydelayincolostrumtransferopensthewaytopredationbypathogenicbacteria.Inthisselectiveframe,femaleshadtoevolutionarilyelaboratemilk-deliveringinterfacesthatbothattractandboostsuckingintheirnewborn,whereasnewbornshadtodetect,process,andadequatelyreacttonipples.Onesensorymeanbywhichnipplesaremadeconspicuoustohumannewbornsisolfaction.Thenipplesoflactatingwomenareindeedendowedwithareolarglands(AG)whichdevelopmentpeaksbytheendofgestationandduringlactation.Theseareolarglands(so-calledMontgomeryglands)arecomposedofsebaceousglandscoalescedwithisolatedlactealacini.Tworeplicatestudies(Schaaletal.,2006;Doucetetal.,2012)haveestablishedarelationshipbetweenthenumberofAG,mother-reportedsuckingoftheinfant,infantweightgainbetweenpostnataldays1and3,andonsetofstage-2lactation.Here,wepresenta3rdreplicatestudytofurtherverifytherelationshipsbetweentheaforementionedvariablesinthe4veryfirstbreast-feeds,withtheassumptionthattheseinauguralfeedsarethosewhichconveythemostprotectiveeffectsonnewborns.

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Amongthe67women(35primiparous)followedup,89.6%had1to15AGoneachareola.TheyallacceptedtocompletetheInfantBreast-FeedingAssessmentTool(IBFAT)afterthefirst4feeds,beforeandafterwhichtheirinfantswereweighed.Theirlactationalphysiologywasfollowedupinassessing3criteriaofincreasedbreastactivity.ItcameoutthatthenumberofAGwaspositivelyrelatedtoreportsofinfantsucking,especiallyinprimiparousmothers.Additionally,womenhavinghigherendowmentsinAGevincedshorterlatencytolactationonsetandtheirinfantstookmoreweightbetweenbirthandday3.ThisstudyevidencessignificantlinksbetweenAGandadaptiveresponsesofneonatesattheirveryfirstcontactwiththelactatingbreast.AGhavingbeenotherwiseshowntoemitsecretionsthatarehighlyreactogenictonewborns(Doucetetal.,2009),wehypothesizethattheseeffectsaremediatedbysomepotentchemicalsignal(s)releasedbyAG.

Talk:PerfumepreferencesinhumancouplesS.C.Roberts1,E.Simpson1,J.Havlicek2,3&C.Allen4

1UniversityofStirling,Stirling,UK;2NationalInstituteofMentalHealth,Klecany,CzechRepublic;3FacultyofScience,CharlesUniversity,Prague,CzechRepublic;4InstituteofNeuroscience,NewcastleUniversity,NewcastleuponTyne,UKSeveralstudiesofhumanmatechoicehaveprovidedevidencefordisassortativeodourpreferences,assometimesoccursinotherspecies,andthatexpressionofthesepreferencesinwomenmaybeinfluencedbyuseofhormonalcontraception.Otherstudiesindicatethatindividualperfumechoicesareinfluencedbytheirinteractionwithone’sownbodyodour,suchthatchoicemaybecriticallyinfluencedbytheperceptionoftheemergentperfume-bodyodourblend.Inlightofthis,wetestedtwopredictionsregardingperfumepreferencesinestablishedheterosexualcouples:(1)thatmembersofthecoupleshouldhavedivergentpreferencesforfragrances,and(2)thatthemagnitudeofwithin-coupledivergenceinpreferenceshouldbereducedbythewoman’suseofhormonalcontraception.Totestthesepredictions,eachmanandwomanin27couplesindependentlyassessedaseriesofeightfragranceaccordsusedinmodernperfumery.Wefoundthat,contrarytoprediction,within-couplepreferencesforfragranceaccordswerepositively,notnegatively,correlated.Concordanceinpreferencewasnotrelatedtothewoman’scontraceptiveuse,eithercurrentlyorpreviouslywhenthecouplemet;however,concordanceinpreferencedidpredictaspectsofrelationshipsatisfactioninwomen,butnottheirmalepartners.Ourresultssuggestthatassortativepreferencesforodoursmaybemoreimportantthanpreviouslybelieved,andthatsharedodourpreferencesmaybelinkedtoaspectsofhumanrelationshipfunctioning.

Talk:Investigatingtheartificiallyscentedape:Theimpactofartificialfragrancesonhumanolfactorycommunication

C.Allen1,J.Havlíček2,3,K.D.Cobey4&S.C.Roberts5

1InstituteofNeuroscience,NewcastleUniversity,NewcastleuponTyne,UK

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2FacultyofScience,CharlesUniversity,Prague,CzechRepublic3NationalInstituteofMentalHealth,Klecany,CzechRepublic4OttawaHospitalResearchInstitute,Ottawa,Canada5UniversityofStirling,Stirling,UKThoughonceconsideredtobefunctionallyanosmicthereisnowagrowingbodyofevidencesupportingtheroleofolfactorycommunicationinhumans.Thisresearchoftenfocuseson‘natural’odours,requiringparticipantstoavoidallfragrancedproductsduringodourcollection.However,wearguethatitisimportanttounderstandhowhumanodoursinteractwithartificialfragrances,andhowthismayaffectolfactorycommunication,giventhatfragrancesarecurrently,andhistorically,ubiquitousacrosssocietiesandcultures.Onestudywillbepresentedwhichinvestigatedtheimpactofartificialfragrancesonourabilitytodiscriminatebetweentheodoursofindividualswhentheywerewearingnofragrance,afragranceoftheirchoice,andanexperimenterassignedfragrance.Ourfindingssuggestthatfragrancechoicemaybeimportant,withdiscriminationratesbeingabovechanceinallconditions,butimportantlywithperformancebeingbetterwhenanindividualwaswearingafragranceofchoicecomparedtoanexperimenterassignedfragrance.Thesecondstudypresentedexaminedtheimpactoffragrancesonourabilitytoaccuratelyassessmatechoicerelevantcuesinbodyodour.Weinvestigatedtheconsistencyofassessmentsbetweenfacesandodours,bothwithandwithoutfragrance,findingthatfragrancesmayenhancecertaincuesinspecificindividuals,potentiallyinterferingwithourabilitytoaccuratelyassessthesecues.Thetwostudiestogetherdemonstratethattheimpactofartificialfragrancesmaybemorecomplexthanoriginallythought,neithersimplymaskingnorenhancingodours,withoutcomesappearingtobedependentonthetypeofinformationbeinginvestigated.

Talk:Chemocommunicationinavision-ruledworldJ.-Y.Baudouin,K.Durand,A.Leleu,A.Seigneuric&B.Schaal

DevelopmentalEthologyandCognitivePsychologyGroup,CenterforSmell,TasteandFoodScience,CNRS-UniversitédeBourgogneFranche-Comté,Dijon,France

Onecanonlyacknowledgethatthehumancommunicationsystemisoverwhelminglydominatedbyvisionand,toalesserextent,byaudition.Thisspecies-typicalsensoryratioledtoastrongbiasinfavorofresearchonvisual/acousticawarenessinenvironmentalandsocialcontexts.Olfactionhasbeenforlongdowngradedtoaminor,relictualsensemodality,thedefectorlossofwhichbeingconsideredasalmostinconsequential.Thelastdecadesofresearchhavenowclearlyshownthatolfactioninfiltratetheregulationofallaspectsofhumanbehaviourandthatanosmiaincursdeepcostsonthequalityofeverydaylife,especiallyinself-adaptationandsocialexchanges.Despitethisrecentconceptualreversal,olfactionremainsmassivelystudiedforitself,inrelativeisolationfromtheothersenses.Thepresenttalkaimstosummarizeseveralstudiesontheeffectsofodorcuesonvariousaspectsofhumanvisualbehaviour.Specificeffectswillbereportedontheimpactoftheodourcontextonthedirectionalityandtemporaldynamicsofvisualbehaviourinsocialand

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non-socialsituationsinbothadultsandinfants.Odorcuescontributetothemultisensoryinterpretationofambiguousvisualcues,inawaytoincreaseaccurateresponses.Insum,theseresultsconfirmthatolfactionhasaclear,althoughsubtle,influenceonvisualprocesses,bringinganotherargumenttodismissthemythwhichobstinatelydefineshumansas“microsmatic”.

Talk:Sexinguphumanpheromones:Howacorporationcreatedamythandleftatrailoffalsepositivespavedbypositivepublicationbias

T.Wyatt

Dept.ofZoology,UniversityofOxford

Acorporationinterestedinpatenting‘humanpheromones’forprofitcreatedalonglastingmyththathasdrawninmanyscientistsaswellasthegeneralpublic.Idescribewhatwentwrongandwhatwouldbeneededtoestablishthatwedohavepheromones(chemicalsignalswithinaspecies).Ashumansaremammals,wemayhavepheromones.However,thereisnorobustbioassay-ledevidenceforthewidelypublishedclaimsthatfoursteroidmoleculesarehumanpheromones:androstenone,androstenol,androstadienone,andestratetraenol.Positiveresultsarehighlylikelytobefalsepositives.Instead,weneedtotaketheleadfromDarwinandtreatourselvesasifwewereanewlydiscoveredmammal,andusetherigorousmethodsalreadyprovensuccessfulinpheromoneresearchonotherspecies.

LunchBreak

Opensession1(Chair:ChristinaDBuesching)Talk:PheromoneofIndianLeopard,Pantherapardus

B.Ghosh1,2,S.Das1,P.Das1,S.Manna1,S.PalChowdhury3,M.PoddarSarkar1&R.L.Bhamachary4

1DepartmentofBotany(CentreofAdvancedStudy),ChemicalsignalandLipidomicslab.,UniversityofCalcutta,Kolkata700019,India;2DepartmentofZoology,BidhannagarCollege,Govt.ofWestBengal,SaltLakeCity,Kolkata700064,India;3DirectorateofForests(WildlifeWing),BikashBhawan,SaltLakeCity,Govt.ofWestBengal,Kolkata700098,India;421B,Flat10,DumdumMotijheel,Kolkata700076,India.

EthochemicalcharacterisationofMarkingFluid(MF)andUrinary(UR)pheromoneoftheIndianLeopard,Pantherapardusfusca(Meyer,1794)werecarriedoutforlastfewyearsonsomewild-bornbutrescuedandrearedundercaptivityintworehabilitationcentresofWestBengal(26˚25ʹ102ʺN,89˚43ʹ33.366ʺEand26˚37ʹ5.142ʺN,89˚12ʹ26.412ʺE),India.Wehighlightthreemodesof'scentmarking'inboththesexesofleopard-directsquirtingwhilestandingerectandthetailisraisedupward(MF),secondsquirtingposturewithlowheight-jetandkeepingkneesbent(MFU)andthethirdonenormalurination(UR)withfoldedlegswhentailalmosttouchestheground.Otherbehaviourslikeflehmen,cheekandbody

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rubbing,nailscratching,vigoroussniffinganddifferentcallswerealsodocumented.HeadspacevolatileswereanalysedfromMFandUrineofbothsexesthroughSolidPhaseMicro-ExtractionguidedGasChromatographyMassSpectrometry(SPME-GCMS).About25compoundswithvariousfunctionalgroupssuchaspyrrole,phenylethylalcohol,benzaldehyde,octanal,methoxy-phenyloxime,acetophenoneetc.wereidentified.TheelusivearomamoleculeofIndianBasmatiricei.e.2-Acetyl-1-Pyrroline(2AP)whichwasidentifiedearlierintigerwasalsodetectedfromvolatilomeofMFoftheanimal.Varioussaturatedandmono-unsaturated,poly-unsaturatedandbranchedchainfattyacidslikehexadecanoic,octadecanoic,docosanoicacid,12-octadecenoic,9,12-octadecadienoic,10-methyldodecanoic,14-methylhexadecanoicacidetc.werealsoidentifiedfromMFandURofLeopard.Thelipidcontainingtriglycerides,waxesters,sterolesters,freefattyacidsetcmightactasfixatives.Theamountoflipidextractedisrangedfrom1-3mg/ml.

Talk:InfantpandascannotdiscriminatebetweenteatodorsofdifferentlactatingfemalesD.Liu1,X.Xie1,H.Wu1,B.Luo2&G.Zhang2

1KeyLaboratoryofBiodiversityScienceandEcologicalEngineeringofMinistryofEducation,CollegeofLifeSciences,BeijingNormalUniversity,Beijing,China;2KeyLaboratoryforReproductionandConservationGeneticsofEndangeredWildlifeofSichuanProvince,ChinaConservationandResearchCenterfortheGiantPanda,Wolong,Sichuan,China

Particularlyinmammals,itisvitalthatneonatescandiscriminatetheirownmotherfromotherfemales.Awealthofstudiesongroup-livingspeciesfromabroadvarietyofanimaltaxaincludinghumansshowsthatinfantsprefertheirmother’sbodyodoroverotherfemales;yet,studiesonsolitaryanimalsareunavailable.Toexamineifgiantpandainfantscandiscriminateteatodorsoflactatingfemales,wefirstusedaparadigmofhabituation-dishabtituationtoinvestigateiftheinfantscandiscriminatetheteatodoroftheirmotherfromthatofanovellactatingfemale.Next,weexaminedifthoseinfantsprefertheirmother’steatodorstothatfromanovellactatingfemaleusingatwo-choicetest.Ourresultsshowedthattheinfantswerehabituatedtotheirmother’steatodorinthehabituationphase(P=0.002,n=7),yetshownosignificantincreasedinterests(investigating,sniffingandplayingwith)intheteatodorofanunfamiliarlactatingfemale(P=0.083,n=7).Duringthetwo-choicetest,thoseinfantsdidnotshowsignificantodorpreferencestoeithersourceofteatodorsoflactatingfemales.Byreferringtoapreviousstudyonthediscriminationbetweenmotheranddaughterafterweaning,weprimarilyconcludethatinfantpandasmaynotbeabletodiscriminatetheteatodoroflactatingfemalekinfromnon-kin.FurtheranalysesofthechemicalcomponentsoftheteatodorbyGC-MSarenecessarytoresolvethispointunambiguously.

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Talk:DecodingMouseSocialCommunication

X.FuXiaoyan1,Y.Yan2,P.Xu1,IL.Geerling-Vidavsky2,M.Gross2&T.Holy1

1DepartmentofNeuroscience,WashingtonUniversityinSt.Louis,USA2DepartmentofChemistry,WashingtonUniversityinSt.Louis,USA

Mouseurineisarichsourceofpheromonesandothersocialodorcueswhichconveyinformationaboutsex,socialstatus,andidentity;however,themolecularnatureofthesecuesisessentiallyunknown.Thevomeronasalorgan(VNO)ofmammalsplaysanessentialroleindetectingcomplexchemicalsignalsthatconveyinformationaboutgender,strain,andthesocialandreproductivestatusofanindividual.Weusedtheresponsesoftheseneuronsasanassaytoidentifyactivecompoundsinurine.Usingaplanararrayof60extracellularmultieletrodestosimultaneouslyrecordsensoryresponsesoflargenumbersofvomeronasalsensoryneurons(VSNs)intheisolatedvomeronasalneuroepithelium(Holyetal,2000),weisolatedthespikingresponsesofindividualVSNsduringinterleavedstimulationwithurinesamplesofbothgendersoffourstrains’(BALB/CJ,CBA/J,C57BL/6J,DBA/2J)mouse.ThesesameurineextractswereprofiledwithnanoLC-MS/MStoquantifytherelativeabundanceoftheindividualmolecularcomponentsacrossdifferenturinesamples.Wedevelopedanewapproach,component-activitymatching(CAM)(Fuetal,2015)toselectcandidateligandsthat‘‘explain’’patternsofbioactivityacrossdiversecomplexmixtures.Usingthisnewapproach,bothpreviously-identified(Nordorietal,2008)andnovelsex-andstrain-specificcompoundswere(Fuetal,2015)orwillbeidentified.Thisapproachprovidesanunprecedentedwaytoacceleratetheidentificationofthemolecularnatureforchemicalcommunicationamongmice.

PMCoffeeBreak

OpenSession2(CarstenTMüller)

Talk:Whatmayattractbluepetrelstotheirburrowsatnight?

M.Gabirot1,F.Bonadonna1&C.T.Müller2

1CEFE-CNRS,Montpellier,France2SchoolofBiosciences,CardiffUniversity,Cardiff,UK

Hypogeanpetrelsreturnduringthesummermonthsonremoteislandstobreed.Thecombinationofnocturnalbehavioursandnestinginthesameburrowcouldallowevolutionofelaborateolfactorysensesinthesepetrels.Theirstrongmuskyodour,theiranatomyandtheirlifestylehaveledauthorstosuggesttheimportantroleofolfactionandplumageinsocialfunctions.Behaviouralexperimentsshowedthatolfactorycuesaresufficienttoallowidentificationoftheirownnest.Achemicalstabilesignaturemightemanatefromburrowandbeinvolvedinnestrecognition.Howevernothingisknownabouttheoriginandthe

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characteristicsoftheseodours.Consequentlyweinvestigatedanddevelopedanovelrobustmethodologyforcapture,analysisandcharacterizationofvolatileorganiccompounds(VOCs)fromnestsofbluepetrels.WecomparedchemicalprofilesfromneststoidentifyVOCskeepingalongyears,originatedfrombirdsornot.Toverifytheimportanceoftheresidentinburrowonnestscents,wealsoextractedVOCsfromfeathers.Despiteofchemicalvariationsamongyearsobserved,nestodourscontainedsocialinformationsuchasspecies,occupancyandnestidentity.Eachnestseemedtohaveasingularchemicalsignature.ImportantlysomeinformativeVOCsfromfeatherswerealsopresentinnestodoursyieldingachemicalsignatureofburrowpotentiallyimplicatedinnestrecognition.Thisworkrepresentsthefirstmultidisciplinarycasestudyofavianchemicalcommunicationusingprincipallyairbornesignals.Theelucidationoftheorigin,natureandfunctionofchemicalcommunicationinbirdshasmajoreco-evolutionaryimplicationsforourunderstandingofavianecology.

Talk:HarderianGlandConstituentsMediateVomeronasalFunctioninginGarterSnakes

E.J.Bentz&R.T.Mason

DepartmentofIntegrativeBiology,OregonStateUniversity,USA

TheHarderianglandisthelargestcephalicglandinmostterrestrialvertebrates,butdespitenumerousstudiesformorethan300years,itsphysiologicalfunctionremainsunresolved.Harderianglandsecretionsarediverseamongdifferenttaxaandmultiplelinesofinquiryhaveresultedinmanyputativefunctionsbeingascribedincludingproductionoflipids,porphyrinsandimmuneproteins.TheHarderianglandsofsquamatereptilesarephysicallyconnectedtothevomeronasalorganandsolelyproducethefluidsinthevomeronasallumen.Proteincomponentsofthesesecretionsareessentialtovomeronasalchemosensoryfunctionfacilitatingthedetectionofsexpheromonesandpreykairomones,chemicalsignalsessentialtomaterecognition,mateselection,andfeeding.Malered-sidedgartersnakes(Thamnophissirtalisparietalis)usetheirvomeronasalorganexclusivelytolocateandevaluatepotentialmatesbasedonfemalesexpheromones,whereasfemalesdonotactivelysearchformates.AnalysisoftheproteincomponentsofgartersnakeHarderianglandsecretionsviaSDS-PAGEshowseveralsecretedproteinswithsexuallydimorphicexpressionpatternslikelytobetargetsofselectionandimportanttothenaturalhistoryofthesesnakes.PreliminaryRNAsequencingresultsshowhighexpressionoflipocalins–proteinswhichbindandsolubilizelipidsinaqueoussolutionlikelyactingaspheromone-bindingproteinsfacilitatingthedetectionofthenonpolarfemalesexualattractivenesspheromone.FurtheranalysesoftheproteincomponentsofHarderianglandsecretionsviamass

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spectrometryandhighthroughputRNAsequencingaimtorevealtheidentityofthesesecretedproteins.

Talk:Effectsoflowenvironmentalcalciumonresponsestofoodandpredator-relatedodoursinfatheadminnows.

R.S.Mirza,Reehan&C.Cornell

DepartmentofBiologyandChemistry,NipissingUniversity,Canada

RecentlyithasbeenfoundinareasofNorthernOntario,WesternEuropeandtheNortheasternUnitedStatesthatCalciumhasbeendecliningduetodecadesofacidraindepositionleachingoutcalciumfromthesoil.Thishasledtodeclinesinzooplanktonspecies,andchangesininvertebratecommunitystructure.Inthisstudy,welookedattheabilityoffatheadminnows,Pimephalespromelas,torespondtofoododours,injury-releasedalarmcuesanddietalarmcuesafterbeingacclimatedtolowcalciumconditions(1mg/L)for7days.Minnowsdidnotincreasesearchactivitytofoododourscomparedtotheblankwatercontrol,butappearedtoshowasearchresponsetobothexperimentalandcontrolodours.Whenexposedtoinjury-releasedalarmcues,minnowsdidnotshowafrightresponsetoinjury-releasedalarmcuesunderlowcalciumconditions.Interestingly,minnowsstillexhibitedafrightresponsetochemicalcuesfromlargemouthbassfedfatheadminnows,butnottocuesfrombassfedkrillunderlowandnormalcalciumconditions(3mg/L).Takentogether,alackofenvironmentalcalciumimpairstheabilityoffatheadminnowstodetectimportantecologicalodours.However,someabilitiesaremaintainedafter7days.Wealsogainsomeinsightintotheuseofcalciumwithinvariousolfactorypathwayswithinfishbrains.

Wednesday30August2017

PLENARY:Underwaterchemistrybetweensalamanders:Evolutionofacourtshippheromonesystem

F.Bossuyt

AmphibianEvolutionLab,BiologyDepartment,VrijeUniversiteitBrussel(VUB),Pleinlaan2,B-1050Brussels,Belgium

Newts,aquaticallyreproducingsalamandersofthefamilySalamandridae,haveevolutionarystronglydiversifiedincourtshipstrategies.Forexample,whilemalesofsomespeciesfirmlyholdthefemaleinanamplexus,othershaveabandonedphysicalcontactaltogetherandorchestratethecourtshipprocessbytail-fanningpheromonestowardsthefemale.These

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pheromonesareproteinsoftheSodefrinPrecursor-likeFactor(SPF)family,whichalreadyhadacourtshipfunctionintheearliestsalamandersabout300millionyearsago.Althoughthepheromonesystemessentiallyretaineditsfunctionofenhancingfemalereceptivitythroughoutsalamanderevolution,SPFproteinsdiversifiedthroughmultiplegeneduplications,andmanynewtsstillco-expressahighphylogeneticdiversityoftheseproteins.HereIwillreviewhowthecombinationoftranscriptomics,proteomicsandmolecularphylogeneticshasproducedinsightsinSPFpheromoneevolution,includingdivergenceatspeciation,evolutioninrelationtochangingcourtshipstrategies,andtheoriginofpheromones.

ThematicSessionII:Proteomics(Chair:SunitaJanssenswillen)Talk:ChallengesinproteomicsoftheMUPmulti-genefamily

R.J.Beynon1,A.J.Davidson2,G.Gomez-Baena1,V.M.Harman1,L.McLean1,M.Prescott1,S.A.Roberts2&J.L.Hurst2

1CentreforProteomeResearch,UniversityofLiverpool,Liverpool,L697ZJ;2MammalianBehaviourandEvolutionGroup,LeahurstCampus,UniversityofLiverpool,CH647TE

Multigenefamilies,suchastheMUPcluster,encodemanyproteinsthatareverysimilarintheirmatureproteinsequences.Thisimposesmultiplechallenges,bothintermsofidentificationandquantificationofdifferentisoformsthatcanbeaddressedbyproteomics.However,manyproteomicsapproachesrequiretheanalysisofpeptidesderivedbyenzymicfragmentationoftargetproteins.InthecaseofMUPs,manysuchpeptidesaresharedbetweenmultipleisoforms,renderingidentificationorquantificationextremelychallenging.Further,theMUPclusterisdynamicandthereisconsiderableevidenceforpreviouslyuncharacterisedproteinsinwild-derivedsamples,renderingtheinbredmousereferencegenomeoflimitedvalue.Inthispresentation,IwilldiscussstrategiesforselectivedeploymentofanalyticalapproachestoisolateandquantifyindividualMUPs,andtoproducearobustanalysisofurinaryMUPpatterncomplexityandstability.Iwillshowhowapproachesonhighresolutionanalysisofintactproteinmassandchargecanrevealdifferencesinexpressionpatternbetweenindividuals,andconfirmthestabilityofsuchpatternsovertime.IwillalsodiscussrationalapproachestoanalysisofMUP-derivedpeptides,andhowcarefulanalysisofthesepeptidescanleadtorobustapproachestoquantificationandtheidentificationofnewMUPs.

Talk:Chemosignalsinanuranbreedingglands

L.M.Schulte,S.Janssenswillen,K.Roelants,S.Matthijs,P.Kok,I.VanBocxlaer&F.Bossuyt

AmphibianEvolutionLab,BiologyDepartment,VrijeUniversiteitBrussel(VUB),Pleinlaan2,B-1050Brussels,Belgium

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Chemicalsignalingisanimportantcomponentofthecourtshipdisplayofmanyaquaticandterrestrialsalamanders.Mostknownpheromonesarepeptidesorproteinswhichareexpressedandsecretedfromspecies-specificmalebreedingglandsduringthereproductiveseasontoattractfemalesortostimulatetheirmatingbehaviours.Becauseanurans(i.e.frogsandtoads)spendalotofenergyinacousticandvisualsignaling,chemicalcommunicationduringcourtshiphasreceivedmuchlessattentioninthisamphibianorder.However,anuransalsohaveawidediversityofsexuallydimorphicglands,whichduetotheirassociationwithpeculiarbreedingbehavioursandtheirhistologicalsimilaritytosalamanderglandshavebeensuggestedtocontaincourtshippheromonesaswell.Becauseofthisgrowingawarenessthatchemosignalsinfrogsmaybemorecommonthancurrentlyappreciated,wecollectedawiderangeofdifferentfrogspeciesduringtheirreproductiveseasons,coveringadiverseselectionofbreedingglandtypesandcombinedhistologywithtranscriptomics,proteomicsandmolecularphylogeneticstosearchforpheromonecandidates.Ouranalysesshowhighlydifferentexpressionpatternsfordifferenttypesofglands,butindicatethatseveralofthemmaybeassociatedwithcourtshipbehavioursincertainspecies.Ourstudythusfurthersupportsaroleforpheromonecontainingbreedingglandsinchemicalcommunicationduringanurancourtship.

Talk:Anewcandidatecourtshippheromoneinaquaticallycourtingnewts.

M.Maex,D.Treer,S.Janssenswillen,I.VanBocxlaer&F.BossuytAmphibianEvolutionLab,BiologyDepartment,VrijeUniversiteitBrussel(VUB),Pleinlaan2,B-1050Brussels,Belgium

Malesofadvancedsalamanders(Salamandroidea)areuniqueamongvertebratesinhavinginternalfertilizationwithoutcopulation.Instead,spermistransferredviaanexternalspermatophore,whichthefemaletakesupwithhercloaca.Sinceasuccessfultransferofspermheavilyreliesonafemalefollowingthemaletrackatspermatophoredeposition,maleshaveevolvedvariouscourtshipdisplaystopersuadeafemaleinpickingupthesperm.Inseveralaquaticallyreproducingnewts,malesdonotlongerdisplayphysicalcontactduringcourtship,butinsteadrelyonemittingchemosignalstothefemale.Previously,weisolatedandexperimentallytestedproteinpheromonesoftheSodefrinPrecursor-likeFactor(SPF)familyanddemonstratedthattheycanelicitfemalecourtshipresponsesinaEuropeannewt.Herewecombinedproteomicanalyseswithtranscriptomeanalyses(RNASeq)ofthepheromone-producingglandtoshowthatseveralEuropeannewtspeciesadditionallysecretemultiple15kDaproteinsinthewaterduringcourtship.Thesecysteine-richproteinsdonotshowanyapparentrelationshiptootherknownproteinsinvertebratesandare,similartoSPF,amongthemosthighlyexpressedproteinsinthepheromone-producingglands.Thishighexpression,togetherwithabundantsecretionduringtail-fanninginwater,suggeststhatthe15kDaproteinsserveacourtshippheromonefunctioninnewts.

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Altogether,ourstudypresentsasetofpromisingpheromonecandidatesthatwillbestudiedinfuturebehaviouralexperiments.Talk:MolecularcomplexityofscentsignalsintheNorwayrat

G.Gómez-Baena1,K.C.Pounder2,J.O.Halstead2,S.D.Armstrong1,M.Prescott1,S.A.Roberts2,L.McLean1,J.L.Hurst2&R.J.Beynon1

1CentreforProteomeResearch,InstituteofIntegrativeBiology,UniversityofLiverpool,CrownStreet,L697ZB,Liverpool,UnitedKingdom;2MammalianBehaviourandEvolutionGroup,UniversityofLiverpool,LeahurstCampus,Neston,UnitedKingdom

Chemicalcommunicationinrodentsismediatedbysignalsdepositedintheenvironmentviaurinaryscentmarks.WeperformedadetailedmolecularcharacterizationofnaturallyvoidedurineanddeliberatescentmarkdepositsinRattusnorvegicus,includingsamplesfromratsofthelaboratorystrainsWistarHanandBrownNorwayandalsowildindividuals.Ourresultsrevealthattheproteincompositionsofraturineandscentmarksdiffersignificantly.Intheurineofmalerats,themainproteincomponentsareisoformsofthemajorurinaryprotein(MUPs)system,afamilyoflipocalinsof~18kDawhichhavebeenshowntoplayacentralroleinchemicalcommunicationinmice.ComprehensiveanalysisoftheMUPsysteminmaleraturineindicatedthatsomeisoformscanbephosphorylated.Inmalescentmarksthemainproteinconstituentsoriginatefromtheventralprostate,notablyprostateinandcystatinrelatedprotein,alongwithacomplexmixtureofN-andC-terminaltruncatedformsofMUPs(~17kDainsize).AnalysisofmaleaccessoryglandstracedtheoriginofthetruncatedMUPstothepreputialglands.Infemales,MUPsareabsentfromurine,butarethemaincomponentsintheproteinfractionoffemalescentmarks.Characterizationofextractsfromclitoralglands,thefemaleequivalentofthemalepreputialglands,identifiedtheseasthesourceofMUPsinfemalescents.TheseMUPsarealsotruncatedattheN-andC-terminalextremities,asfoundinmalescents.OurdatarevealanunexpectedcomplexityinthemolecularcompositionofscentmarksinRattusnorvegicus.ThisgivesrisetonewhypothesesabouttheroleofthedifferentcomponentsinchemicalcommunicationintheNorwayrat.

AMCoffeeBreakTalk:Impactofmaleeffectandanoestrusoestrusstatusesongoatolfactorysecretome

M.Chabi¹²,C.LeDanvic³,M.Chasles¹,M.Keller¹,P.Nagnan-LeMeillour²

¹UMRPRCINRA;²UGSF-UMR8576-USCINRA;³ALLICE

Itisreportedinungulates(goatandsheep)thatanactivemaleputincontactwithanoestrusfemalescaninduceaquickchangeinfemaleLHsecretion,whichtriggersfemalesexual

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activityreactivation(Chaslesetal.2016).This“maleeffect”canbemimicbymaleodours.Thosechemicalsignalsaredetectedbythefemalethankstoodorantreceptors(OR).TobringodorantstotheOR,Odorant-BindingProteinsarerequired(OBP).OBPsaresmallproteinssecretedinnasalmucusproposedtoinsureodorantsolubilisationandtogiveitsrecognitionspecificityforthedifferentanimalspecies.Thoseproteinshavebeenshowntobearpost-translationalmodifications(PTM)includingO-GlcNAcylationandphosphorylationinpig,forthefirsttimein2014(Nagnan-LeMeillouretal.,2014).Unlikepigs,goatsdisplayaseasonalreproductiveactivity,andthusgettwostatuses:anoestrusandoestrusbutarethosestatusescharacterizedbyspecificOBPmodification?Inaddition,ifthemaleodourgetsthecapacitytoinduceLHsecretioninfemale,doesthemaleeffectinduceanyPTMchangesonOBPsingoats?Toanswerthesequestions,weinvestigatedforthefirsttimethegoatolfactorysecretomeatdifferentreproductionstatuses.Weused2D-DIGE(2-dimensionaldifferentialelectrophoresis)coupledtohigh-resolutionmassspectrometrytocompareolfactorysecretomesofanoestrus,oestrusfemalesandfemalesexposedtoanactivemalebouquet.ThosemethodsallowedustodescribeforthefirsttimethegoatOBPsinvolvedinthereproductionprocessandhighlighttheputativePTMimplication.Talk:Identificationandcharacterizationofaseasonally-expressedsexuallydimorphicurinarylipocalininthebankvoleMyodesglareolus

G.M.Loxley1,J.Unsworth1,M.Turton1,A.Davidson2,A.Jebb2,J.L.Hurst2&R.J.Beynon1

¹CentreforProteomeResearch,InstituteofIntegrativeBiology,UniversityofLiverpool,UK;²MammalianBehaviourandEvolutionGroup,InstituteofIntegrativeBiology,UniversityofLiverpool,UK

Theexpressionofhighlevelsofproteininurineiscommoninsomegroupsofrodents,providingimportantsignalsinscentaswellasprovidingaslowreleaseofvolatileligandsboundtotheprotein.Rats&mice(muroidrodents)releaseabundantlevelsofMUPsthatexhibithighlevelsofpolymorphism.Voles(cricetidrodents)ontheotherhanddisplaysimplerproteinprofiles,butthepredominantproteinsarelipocalinsrelatedtoMUPs,genericallytermedodorantbindingproteins.Inthisstudy,wecharacterizedaseasonally-expressed,male-specificurinarylipocalininthebankvole,Myodesglareolus.Theproteinsequence,obtainedbypeptidemassspectrometrypredictedaproteinthatwasexactlyconsonantwiththeobservedmassoftheprotein.Further,leucine/isoleucineambiguitywasresolvedbymetaboliclabelling,withdietarydeuteratedleucine.Theproteinisalipocalin,mostcloselyrelatedtoaphrodisinandodorantbindingproteins(OBPs),withaclearlydefinedcentralcavity.However,itisdistinctfromthreeurinaryproteinspreviouslyidentifiedinthisspecies,whicharepresentatmuchlowerlevels.Homologymodellingrevealedtwoequallyvalidconformationsthatalteredtheaccessibilityofthecentralbindingcavity,suggestingaroleforligandbindingandrelease.Thepresenceoftheswitchable

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bindingpocket,combinedwiththeseasonalandmale-specificexpressionpatternleadstothepresumptionthatthisproteinisinvolvedinsexualchemicalcommunicationinbankvoles.Talk:Apreliminarystudyontheanalsacproteomeindogs(Canisfamiliaris)

S.Janssenswillen1,M.Metzemaekers2,P.Proost2&F.Bossuyt1

1AmphibianEvolutionLab,BiologyDepartment,VrijeUniversiteitBrussel,Pleinlaan2,B-1050Brussels,Belgium;2RegaInstitute,LaboratoryofMolecularImmunology,KatholiekeUniversiteitLeuven,Herestraat49–Bus1042,B-3000Leuven,Belgium

Dogsmarktheirenvironmentwithscentsignalstoinformconspecificsabouttheirindividualstatus.Analsacglandsareknowntoplayanimportantroleduringthischemicalsignalling.Sofar,smallvolatilemoleculesandbacterialflorahavebeenidentifiedfromtheseglandsecretions.However,analsacsalsosecretelargequantitiesofnonvolatileproteins,whichuntilnowhaveremaineduncharacterised.Hereweexaminedtheanalsacsecretionsoftwentydomesticdogs.Toinvestigatewhetheranindividual’sstatusisreflectedintheanalsacproteome,excretionpatternswerecomparedamonggenders,ages,breeds,andheatcycletimes.Additionally,weassessedtheimpactofneuteringonscentmarking.Comparativeproteomicsandsequencingshoweddifferentialexcretionofa17kDaproteinthatweidentifiedasanOdorantBindingProtein(OBP),amemberofthelipocalinproteinfamily.Inothervertebrates,OBPsserveaspheromonesandasodorant/pheromonecarriers.Screeningofthedoggenomerevealsthatthecorrespondingobpgeneispartofaclusterofthreerelatedgenes,oneofwhichencodesCanf4,anOBPpreviouslyfoundtobeabundantinsalivaanddander.WefoundthattheanalsacOBPreacheshighconcentrationsinpuppiesandyoungdogs,butnotinadults.Thisfindingsuggeststhatthisproteinhasafunctioninchemicalcommunicationduringthefirstyearsofadog’slife,whichmaybelostwhenraisedinhouseholdsituations.OurpreliminarydatashowthatproteomeanalysescanprovidevaluableinformationforfurtherunravellingthechemicalmechanismsofscentsignallinginCanidae.PostertalksI:ProteomicsPOSTER1:Beyondsodefrin:evidenceforamulti-componentpheromonesysteminthemodelnewtCynopspyrrhogaster(Salamandridae)

I.VanBocxlaer1,M.Maex1,D.Treer1,S.Janssenswillen1,R.Janssens2,W.Vandebergh1,P.Proost2&F.Bossuyt1

1AmphibianEvolutionLab,BiologyDepartment,VrijeUniversiteitBrussel(VUB),Pleinlaan2,B-1050Brussels,Belgium;

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2LaboratoryofMolecularImmunology,DepartmentofMicrobiologyandImmunology,KULeuven-UniversityofLeuven,Minderbroedersstraat10-box1030,B-3000Leuven,Belgium

Sodefrin,adecapeptideisolatedfromthemaledorsalglandoftheJapanesefirebellynewtCynopspyrrhogaster,wasthefirstpeptidepheromoneidentifiedfromavertebrate.Thefirebellysalamanderandsodefrinhavebecomeamodelforsexpheromoneinvestigationinaquaticallycourtingsalamanderseversince.SubsequentstudiesinothersalamandersidentifiedSPFproteincourtshippheromonesofaround20 kDabelongingtothesamegene-family.AlthoughtranscriptsoftheseproteinscouldbePCR-amplifiedinCynops,itiscurrentlyunknownwhethertheyeffectivelyusefull-lengthSPFpheromonesnexttosodefrin.Herewecombinedtranscriptomics,proteomicsandphylogeneticstoinvestigateSPFpheromoneuseinCynopspyrrhogaster.Ourdatashowthatnotsodefrintranscripts,butmultipleSPFtranscriptsmakeupthemajorityoftheexpressionprofileinthedorsalglandofthisnewt.Proteomeanalysesofwaterinwhichamalehasbeencourtingconfirmthatthisproteinblendiseffectivelysecretedandtail-fannedtothefemale.Bycombiningphylogeneticsandexpressiondata,weshowthatindependentevolutionarylineagesoftheseSPF’swerealreadyexpressedinancestralCynopsspeciesbeforetheoriginofsodefrin.ExtantCynopsspeciescontinuetousethismulti-componentpheromonesystem,consistingofvariousproteinsinadditiontoalineage-specificpeptide.POSTER2:Seasonalchangesinolfactoryproteinsofewe

P.Cann¹,M.Chabi¹,C.LeDanvic²,R.LamiR³,M.Keller⁴&P.Nagnan-LeMeillour¹

1UMR8576CNRS/UniversitédeLille1/USCINRA1409,France;2ALLICE,USCINRAUGSF;3LBBM-USR3579CNRS–UPMC;4UMR7247INRA/CNRS/UniversitédeTours

Sheepspeciesdisplaysaseasonalbreedingcharacterizedbyaperiodofsexualactivityfollowedbyaperiodofsexualrest.Duringthefemaleanoestrus,theperceptionoftheodourofasexuallyactiveraminducesreactivationofewegonadotropicaxis,leadingtooestrus.Changesinolfactorysystemhavebeenreportedaccordingtothisswitch,butmorelikelyatcentralnervousstepsthanatperipherallevelofodoursdetection,partlyrealisedbyodorant-bindingproteins(OBP).Wesuggestthatinsheep,theolfactorysecretomeismainlycomposedofOBPisoforms,asitwasdemonstratedinpigspecies(Nagnan-LeMeillouretal.,2014),andthatitscompositioncouldvaryaccordingtothephysiologicalstatusoftheanimal.Besides,ithasbeenshownthatthepresenceofbacteriadecreasestheleveloftranscriptsencodingOBPsinrat(Françoisetal.,2016).So,theaimofourstudyistocompareeweolfactorysecretomeandbacteriomebetweentheanoestrusandoestrusperiods.Proteinsextractedfromnasalmucusoftwoeweswereanalysedby2-dimensionalelectrophoresisfollowedbyMassSpectrometryanalysis(MALDI-TOF/TOFandNano-LC-MS/MS)andcomparedbetweenthetwosexualperiods.Wereportherethefirstidentificationofeweolfactorysecretomeanditsvariationbetweenoestrusandanoestrusperiods.Ourresultscomfortthehypothesisthatthesensoryequipmentofeweisunder

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controlofitsphysiologicalstatus,someproteinsbeingspecificallyexpressedatoestrustime,possiblytoincreasethedetectionofsexuallyactiveramodour.POSTER3:Courtshippheromonesinmudpuppiesandwaterdogs(Necturus,Proteidae)

D.Treer1,M.Maex1,M.A.Stoops2,A.Nagel2,C.J.DeChant3,I.VanBocxlaer1&F.Bossuyt1

1AmphibianEvolutionLab,BiologyDepartment,VrijeUniversiteitBrussel(VUB),Pleinlaan2,B-1050Brussels,Belgium;2CenterforConservationandResearchofEndangeredWildlife(CREW),CincinnatiZoo&BotanicalGarden,3400VineStreet,Cincinnati,Ohio45220,USA;3InnovativeZoologicalSolutions,Cincinnati,Ohio45205,USA

Mudpuppiesandwaterdogs(genusNecturus)arerelativelybig,paedomorphicsalamandersthatinhabitriversandlakesofNorthAmerica.Togetherwiththeolms(Proteusanguinus),theirblindEuropeanrelatives,theyaretheonlyextantrepresentativesofthefamilyProteidae.Necturusspeciesarecharacterisedbylongevityandaprolongedtimeforreachingsexualmaturity.Theyreproducebyinternalfertilisation,inwhichfemalespickupspermatophoresdepositedbythemalesintheenvironment.Becauseseveralinternallyfertilisingsalamanderfamiliesareknowntousepheromonecommunicationtooptimisetheirreproductivestrategy,westudiedthecourtshipbehaviourofNecturusspeciestoidentifybehaviouralpatternsrelatedtopheromonetransfer.Ourobservationsrevealedthatbothsexeswagtheirtailinproximityofanindividualoftheoppositesex.Thisbehaviour,incombinationwiththefactthatthesespeciespossesscloacalglandsthatarehomologoustopheromoneglandsofothersalamanderfamilies,indicatesthatchemicalcommunicationplaysanimportantroleduringcourtship.Wethencombinedtranscriptomicsofthepheromoneglands,proteomicsofcourtshipwaterandmolecularphylogeneticanalysestofurtheridentifycandidatepheromonesinthisurodelanlineage.Ourstudyfillsanimportantgapofinformationonpheromoneuseinanapproximately150millionyearoldevolutionarylineageofsalamanders.POSTER4:Anon-conventionalglycosylationisinvolvedinthecodingofspecificodorsindomesticpig

A.Joly,S.Potelle,C.Spriet,C.LeDanvic,P.NagnanLeMeillour

UnitédeGlycobiologieStructuraleetFonctionnelle(UGSF),UMR8576,CNRS/UniversitédeLille1/USCINRA1409citéscientifique,BâtimentC975655Villeneuved'ascqCEDEX

Inmammals,OdorantBindingProteins(OBPs)aresmallsolubleandsecretedproteinsthatbindandsolubilizeodoursandpheromones,priortointeractionwitholfactoryreceptors(ORs).ThenatureoftheligandofORsisstillunknownandcouldbeeithertheodorantcompounditselforthecomplexformedbythebindingbetweentheOBPandtheodorant.ThelaterhypothesisimpliesalargerOBPdiversitythanthe2-3geneproductsidentifiedineachspecies.Nevertheless,weidentifiedaboutthirtyisoformsofOBPinthepigolfactorysecretomebytwo-dimensionalelectrophoresiscoupledtohigh-resolutionmass

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spectrometry(Nagnan-LeMeillouretal.,2014).Theseisoformsaregeneratedbypost-translationalmodifications(PTMs):phosphorylationandO-b-N-acetylglucosaminylation(O-GlcNAcylation).ThislaterPTMisusuallydescribedfornuclearandcytoplasmicproteinswheretheadditionofGlcNAconserineandthreonineresiduesiscatalyzedbyasingleO-GlcNActransferase(OGT).However,OBPsaresecretedproteinsforwhichthereareonlyafewreportsofextracellularO-GlcNAcylation.Recently,anenzymecalledEGF-domainrepeats-N-acetylglucosaminyltransferase(EOGT)wasevidencedinDrosophilatoperformO-GlcNAcylationofextracellulardomainsoftransmembraneproteins.TofigureoutifO-GlcNAcylationofporcine(Susscrofa)OBPiscatalyzedbyporcineEOGT(Nagnan-LeMeillouretal.,2014),weproducedEOGTintwodifferentexpressionsystemstoanalyzeitsenzymaticactivityinvitrowithitstwoputativesubstrates,GlcNAcandOBP.WealsoperformedanincelluloanalysisofOBPO-GlcNAcylationbyagenerescuemethodinHeLacellline.WealsomonitoredEOGTtraffickingbyalivecell-screeningsystem.

LunchBreak

ThematicSessionIII:OlfactorySpeciationTalk:Comparativeproteinstructurepredictionandelucidationofbovinevomeronasal1receptortype-I(VN1R1)adoptingin-silicostudy

R.Durairaj,C.Bienboire-Frosini&P.Pageat

ResearchInstituteinSemiochemistryandAppliedEthology

Theperceptionofputativepheromoneswasmediatedbyvomeronasal1receptors(V1R)ofG-proteincoupledreceptor(GPCR)superfamilyproteinsintheaccessoryolfactoryepithelium(AOE).Also,theV1Rlocatedinthenasalcavityandinterconnectinglargeolfactorysensoryneuronsdirectedbymultigenesuperfamilyinmostspecies.Inbovine,theV1Riscruciallyinvolvedinchemicalcommunicationrelatedtoreproductionandpheromonetransportactivity.However,thestructuremodelingandannotationofbovineV1Rhasnotyetbeenreported.Inthispresentstudy,thesequenceanalysis,structurepredictionandfunctionelucidationofbovineV1Rmadeusingcomputationalmethods.Theresultsrevealedthat:(i)thehighestproportionofα-helix,strand,andrandomcoilinsecondarystructureofV1R;(ii)ComparativestructurepredictionofV1RwithvarioushomologtemplatesusingAbinitiomodeling;(iii)Validationofstructureshowed98.6%ofmostfavouredregionswithallowedregions;(iv)Structuralsuperimposition,topologyandtransmembranepredictionwereadditionallysupportedthestructuralmodelingofbovineV1Rusingcomputationalanalysis.Thus,forthefirsttimedemonstratedthecomparativestructurepredictionofbovineV1Rfacilitatetopredicttheputativeisoformsofpheromonesinvolvedsignaltransductioninreproduction.

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Talk:Anovellipidmetabolismsystemdevelopedincatsproducesaspecies-specificchemosignal

M.Miyazaki,A.Futsuta,W.Hojo,T.Miyazaki&T.Yamashita

IwateUniversity,JAPAN

Manymammalssecretespecies-specificchemicalsthatfunctionassignalsforconspecificrecognition.Previousstudiesidentifiedspecies-specificchemicalsinseveralspecies.Forexample,domesticcatsexcrete2-amino-7-hydroxy-5,5-dimethyl-4-thiaheptanoicacidknownasfelinineincaturine.Felinineisaprecursorofvolatile3-mercapto-3-methylbutanol,whichisamajorcontributortocaturinaryodor.However,littleisknownregardinghowcatsevolutionallyacquiredtheirbiosyntheticsystems.Hereweshowapreviouslyunknownmetabolicpathwaythatdevelopsindomesticcatsandproducesacat-specificchemosignal.Theincreasedisomeraselevelsinthemevalonatepathwayforisoprenoid/cholesterolbiosynthesisdevelopauniquepathwayfromacholesterolbiosynthesisintermediatetoelectrophilicalcohol,whichisconjugatedtoglutathione.Thenovelglutathione-conjugateisultimatelymetabolizedtofelinineinthecaturine.Sincethemodifiedmevalonatepathwayregulatestheglutathione-conjugateanditsmetabolitelevels,butnotserumcholesterollevelsincatsfedahigh-fatdiet,thismodificationexcelstopreventbiosynthesisofexcesscholesterolinobligatecarnivorecats.Thesefindingsimproveourunderstandingofboththelipidmetabolismofobligatecarnivoresandthebiosyntheticmechanismsofspecies-specificcompoundsforscentcommunication.Talk:OlfactoryspeciationinEurasianbadgersMelesspp.:Diversityofsubcaudalchemicalsandbehaviouralexperiments

Y.Kaneko,Yayoi1,C.Newman2,E.Raichev3,S.Kosuga4,S.Yachimori5,M.Saito1,Y.Kuramoto6,D.W.Macdonald2,S.Peeva3,C.D.Buesching2

1CARNECCO,TokyoUniversityofAgricultureandTechnology;2WildCRU,UniversityofOxford;3TrakiaUniversity;4DaidoBunsekiResearchInc;5ShikokuInstituteofNaturalHistory;6JapaneseMinistryoftheEnvironment

Evolutionarytheorypredictsmembersofdistinct(sub-)speciestobeabletorecognizeeachother,resultinginspeciation.However,surprisinglyfewstudiesattempttotestthistheory.Here,weusebadgers(Melesspp.)asamodeltoinvestigateiftheirodourprofilesreflectpopulation-andspeciesdifferencesbetween2populationsofJapanese,M.anakuma,andEuropeanbadgers,M.meles.Allbadgerspossessasubcaudalgland,uniquetothisgenus.WepositthatthegeneralcompositionofbadgerMelesspp.secretionsremainsimilarovergeographicdistancesandevolutionarytime,encodinggenus,butthatodour-profilesbetweenM.melesandM.anakumadiffersufficientlytoallowspeciesdiscrimination,and

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geographically-discretepopulationsofeachspecieswillvarywithinspecies-specificparameters.Weanalysed41adultbadgersub-caudalglandGC-MSprofilesfrom2M.meles(UK,Bulgaria)and2M.anakuma(Tokyo,Kochi)populations.Between24�32chemicalcomponentswereidentifiedineachprofile,4compoundswerepresentineachindividual,and10compoundswerepresentinbothspecies.Statisticalanalysesrevealedsignificantdifferencesbetweenthetwospecies(PERMANOVA,p<0.05)aswellasbetweensubpopulationsofeachspecies(PERMANOVA,p<0.05).Behaviouralscent-provisioningexperimentsonacaptiveadultfemaleJapanesebadgerrevealedthatthesestatisticaldifferencesarebiologicallyrelevantwiththeanimalsnififfingscentfromotherM.anakumasignificantlymoreoftenthanscentfromM.meles.Talk:OdourdialectsamongwildmammalsE.F.Kean,M.W.Bruford,I.M.Russo,C.T.Müller&E.A.Chadwick

SchoolofBiosciences,CardiffUniversity,Cardiff,UK

Acrossmultipletaxa,populationstructureanddynamicsdependoneffectivesignallingbetweenindividuals.Amongmammals,chemicalcommunicationisarguablythemostimportantsense,underpinningmatechoice,parentalcare,territorialityandevendiseasetransmission.Thereisagrowingbodyofevidencethatodoursignalscarrygeneticinformationthatmayconferconsiderablebenefitsincludinginbreedingavoidanceandnepotism.Todate,however,therehasbeennoclearevidencethatodourssignalpopulation-levelinformationinwildmammals.Herewedemonstrateforthefirsttimetheexistenceof'odourdialects'ingeneticallydistinctmammaliansubpopulationsacrossalargegeographicalscale.Wefoundthatotters,Lutralutra,fromacrosstheUnitedKingdompossesssexandbiogeography-specificodours.Subpopulationswiththemostdistinctiveodourprofilesarealsothemostgeneticallydiversebutnotthemostgeneticallydifferentiated.Furthermore,geographicdistancebetweenindividualsdoesnotexplainregionalodourdifferences,refutingotherpotentialexplanationssuchasgroupodoursharingbehaviour.Differencesinthelanguageofodoursbetweensubpopulationshavethepotentialtoaffectindividualinteractions,withclearimplicationsforreproductionandgene-flow.Odourdialectsthusprovideapotentiallyimportantmechanismgoverningmammalianmeta-populationstructureacrosslargelandscapes.

PMCoffeeBreak

Postertalks2:Proteomics&PrimateolfactionPOSTER5:QuantitativeProfilingofMajorUrinaryProteinsinWildMice

M.C.Prescott1,G.Gómez-Baena1,V.M.Harman1,A.J.Davidson2,S.A.Roberts2,J.L.Hurst2&R.J.Beynon1

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1CentreforProteomeResearch,InstituteofIntegrativeBiology,UniversityofLiverpool,UK;2MammalianBehaviourandEvolutionGroup,InstituteofIntegrativeBiology,UniversityofLiverpool,UK

TheMajorUrinaryProteins(MUPs)arebeta-barrellipocalinsof18-19kDathatareinvolvedinrodentcommunication.MUPsencodeowneridentityandelicitthetransportandslowreleaseofvolatilemolecules.OneMUP,darcin,isamalepheromonethatinduceslongtermrememberedattractiontotheownerofascentmarkandhislocation.IndividualwildmiceexpressdifferentMUPprofiles,requiringrapidanalyticalapproachestoobtainfirst-passprofilingofMUPcontent.TheprofileofMUPscanbeanalysedintwophysicochemicaldimensionsusingmassspectrometry(MS)andisoelectricfocusing(IEF).Analysisoftheelectrosprayionisation(ESI)massspectrumofintactMUPsdefinesthepatternofproteinsseparatedbydifferencesinmassandreturnsarelativequantificationofthecomponents.Forabsolutequantificationinmassspectrometry,standardsarerequired.Wehaveusedsyntheticproteinstandards(QconCATs)thatareproducedbyartificialgenedesign.TheQconCATsareproducedinbacteriaandsimultaneouslylabelledwithstableisotopes.AddingQconCATStoanaturalMUPmixture,followedbydigestionusingendopeptidases,co-generatesquantotypicpeptidestandards(Q-peptides)alongsidenaturallyderivedanalytepeptides.Whenanalysedbyliquidchromatography-massspectrometry(LC-MS)absolutequantificationofMUPsincomplexsamplescanbeachievedbyreferencetothelabelled,mass-shiftedstandards.WehavealsoevaluatedanautomatedIEFsystem(ProteinSimplenanoPro1000)asanorthogonalmethodforquantitativeMUPanalysis,withspecificdetectionusingaMUPantibody.WewillpresentexamplesoftheapplicationofintactmassESI-MS,QconCATwithLC-MSandIEFforquantitativeMUPanalysisinwildmice.POSTER6:Identificationandcharacterizationofaseasonally-expressedsexuallydimorphicurinaryproteinintwospeciesofMalagasymouselemurs,MicrocebusmurinusandMicrocebuslehilahytsara

G.Loxley1,A.Trester2,J.Unsworth1,G.Gomez-Baena1,U.Radespiel2,E.Zimmermann2,J.L.Hurst1,R.J.Beynon1

1InstituteofIntegrativeBiology,UniversityofLiverpool,UK;2InstituteofZoology,UniversityofVeterinaryMedicine(TiHo),Hannover,Germany

Communicationviasensorycuesgovernsreproductionandsurvivalinprimates.MalagasylemursbelongtothebasalprimategroupStrepsirrhini,exhibitingarhinariumandafunctionalvomeronasalorgan,supportingimportanceofchemosignallinginsocialcommunication.Lemursdisplayaseasonalreproductionpattern.Urinaryproteinsplayanincreasingnumberofrolesincommunicationinrodents.Proteinuriainprimatesisusuallythoughttobeasignofrenaldysfunction.Ouraimwastoexplorewhetherprimateurinaryproteinsarelinkedtosex,seasonandreproduction,andmaythuspotentiallybeinvolvedinchemosignalling.UrinesamplesofMalagasymouselemurswerecollectedinandoutofthebreedingseason,frommaleandfemaleadults.Anabundantproteinwasevidentinsomemaleurinesamples,butlimitedtothebreedingseason.Furthercharacterizationidentified

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thisasaWAPfour-disulphidecoredomainprotein12(WFDC12),homologoustoanti-microbialandanti-peptidaseproteins.Thisproteinwasquantifiableinindividualsfromallgroups;inbreedingseasonmales,however,therewasa70-foldaverageincreaseinabundancecomparedtoallothermales.Astrongsex-specificdifferencewasvisible,witha90-foldincreaseeveninlow-WFDC12malesinthebreedingseasoncomparedtoallfemales.Anumberofotherproteinsrevealedsimilarincreasesinabundanceinthissamesubsetofmalesfromtheirhomologoussequencesinotherprimates,includingsemenogelinII,aseminalvesicleproteinthatincreasedonaverageby4000-fold.ThisrelationshipbetweenanincreasedexpressionofsemenogelinIIandWFDC12remainstobedefined.Weestablishbio-essaystoassessanysemiochemicalroleoftheWFDC12protein.

POSTER7:Makingsenseofscents:DecipheringprimateolfactorycommunicationA.Poirier

AngliaRuskinUniversity,Cambridge,UK

Olfactorycommunicationisanimportantmediatorofsocialinteractionsinmammals,providinginformationaboutanindividual’sidentityandcurrentsocial,reproductive,andhealthstatus.Thisresearchprojectuniquelycombinesbehaviouralandchemicalinformation,inbothwildandcaptiveconditions,todeciphersomeaspectsofolfactorycommunicationincallitrichines(marmosetsandtamarins,NewWorldprimates).Thestudyexploreshowspecies,social,andindividualcharacteristics,butalsoenvironmentalconditions(i.e.captivevs.wildconditions),mightinfluencepatternsofcallitrichinescent-markingbehaviour,aswellasthechemicalcompositionofscent-marksecretions.Behaviouralobservations,aswellasswabsofscent-glandsandnaturallydepositedscent-marks,werecollectedfromcaptivegroupsofmarmosetsandtamarinsinseveralUKzoos.Additionally,swabsofscent-glandswereobtainedfromawildpopulationoftamarinsintheSouth-easternPeruvianAmazonaspartofanannualtrap-and-releaseprogram,andcomparedwiththosecollectedincaptiveconditions.Chemicalsampleswereanalysedusingheadspacegaschromatography-massspectrometry.Thisstudyintendstoprovidebaselineknowledgeapplicabletocaptivehusbandrypractices,includingbreedingofrarespecies.POSTER8:EffectsofMaleAxillaryExtractsontheLengthofMenstrualCycleinReproductiveAgeWomenandPre-MenopausalWomen

V.V.Voznessenskaya&T.K.Laktionova

A.N.SevertsovInstituteofEcology&Evolution,RussianAcademyofSciences

Humanunderarmsecretions,whenappliedtowomenrecipients,alterthelengthandtimingofthemenstrualcycle(Pretietal.,2003).Specificaimofourstudywastoinvestigatetheinfluenceofmaleaxillaryextractsonthelengthofmenstrualcycleofpremenopausalwomenincomparisonwithwomenofreproductiveage.Totalof43women,ageof21-51,participatedinourstudy.Wemonitoredlengthofthemenstrualcycleforeachtestsubject

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for4monthsbeforeexperimentandfollowedmonitoringfor2monthsaftertheexperiment.Weappliedaxillaryextracts/ordiluentontheupperlip3timesaweekfor6hoursinthemorningfordurationof9weeks.Womenof21-44yearsoldweresubdividedintothreegroups:withmenstrualcyclelengthof26-32days(1);menstrualcyclesshorterthan26days(2)andwithcycles>32days(3).Datafromwomenof>45yearsoldwereanalyzedseparately.Maleaxillaryextracts(MAE)didnotaffectsignificantlylengthandregularityofmenstrualcycleinwomenwithnormalandregularcycles(n=12,p>0.1).Atthesametimeweobservedsignificantlyshortermenstrualcycles(n=12,p<0.01)underMAEapplicationsinwomenwithcycleslongerthat32days.Forwomenwithmenstrualcyclelength<26dayswealsodidnotobservestatisticallysignificantchangesunderMAEtreatment(n=9).InpremenopausalwomenMAEapplicationscausedsignificantshorteningofthemenstrualcycle(n=10,p<0.01).Alsoweobservedatendencyformoreregularcyclesforthisgroupoftestsubjects.SupportedbyRSF16-15-10312toVVV.

OpenSession3

Talk:Socialodorspromotefood-relatedpreferencelearningthroughactivationoftheGC-D/necklaceolfactorysubsystem

A.D.Zimmerman1,2,H.Zhi1,2&S.D.Munger1,2,3

1CenterforSmellandTaste2DepartmentofPharmacology&Therapeutics3DepartmentofMedicine,DivisionofEndocrinology,DiabetesandNutrition,UniversityofFlorida,Gainesville,FL,32610,USA

Individualsfromseveraldiversemammalianspeciescantransmitdietarypreferencestopeerconspecificsandhelpanimalstoavoidpotentiallytoxicfoods.This“socialtransmissionoffoodpreference”(STFP)requiresconcurrentdetectionofoneofseveralsemiochemicals(CS2presentinthebreath,orguanylinpeptidesexcretedinfecesorurine)andodorsassociatedwithaparticularfoodsource.Thesesemiochemicalsstimulateaspecializedolfactorysubsysteminmicethatincludeolfactorysensoryneurons(OSNs)expressingtheuniqueolfactoryreceptorGC-D(guanylyl-cyclaseD).GC-D+OSNsprojecttothecaudalmainolfactorybulb,apoorlydefinedregionthatincludesthe“necklaceglomeruli”(atargetofGC-D+OSNs)andthatalsoreceivesinputfromotherchemosensoryneuronsthatutilizecGMP-mediatedtransductionmechanisms.RecentpublishedandunpublishedstudiesofSTFPinmicewillbediscussed,includingadissectionofthemolecularmechanismsemployedbyGC-D+OSNstodetectCS2andguanylinpeptides;tracttracingandactivitymappingofneuralcircuitsassociatedwiththeGC-D/necklacesubsystem;andbehavioralanalysesofolfactorysamplingandconsumptioninthecontextofthisassociativelearningparadigm.SupportedbyNIDCDgrantDC005633.

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Talk:Influencesofexposuretomurinepheromoneonthenextgeneration

S.Koyama1,H.Soini2,U.Tadasu3&M.Novotny2

1IndianaUniversity,MedicalSciences,Bloomington,IN,U.S.A;2IndianaUniversity,Dept.Chem.,Bloomington,INU.S.A;3ObihiroUniversityofAgricultureandVeterinaryMedicine

Thecurrentunderstandingoftheactivityofmammalianpheromonesisthatendocrineandbehaviouraleffectsarelimitedtotheexposedindividuals.Here,wedemonstratethatthenasalexposureoffemalemicetoamalemurinepheromonestimulatesexpansionofmammaryglands,leadingtoprolongednursingofpups.Subsequentbehaviouraltestingofthepupsfrompheromone-exposeddamsexhibitedenhancedlearning.Sialicacidcomponentsinthemilkareknowntobeinvolvedinbraindevelopment.Wehypothesizedthattheoffspringmighthavereceivedmoreofthiskeynutrientthatpromotesbraindevelopment.ThemRNAforpolysialyltransferase,whichproducespolysialylatedneuralcelladhesionmoleculerelatedtobraindevelopment,wasincreasedinthebrainofoffspringofpheromone-exposeddamsatpost-natalday10whileitwasnotdifferentatembryonicstages,indicatingpossibledifferentialbraindevelopmentduringearlypost-natallife.Wecollectedmilkfromdamsatpost-deliveryday10,analyzedthemilkusingHPLCand1H-NMRMALDI-TOFMS.Wefoundthattherearedifferencesintheamountofsialicacidinthemilkofpheromoneexposedfemalescomparedtocontrols(femalesexposedtowaterandfemaleswithnotreatment).

Thursday31August2017

PLENARY:Domammalshaveaworldwidewebofinterspecificscentsignals,andcouldweconnecttoit?

P.Apps

BotswanaPredatorConservationTrust

Althoughnearlyalltheworkonmammalsemiochemistryhasbeenonsinglespecies,interspecificscentmarkingisquitewidespread,andrecentcameratrappingstudiesofcarnivoreguildsinCanada,thePacificNorthwestoftheUSA,CostaRicaandBotswanahaveshowncarnivoresusingmulti-speciesscent-markingsites.Therearerecordsofinterspecificmarkingfromatleast44mammals,witharapidlyexpandingwebofknownconnectionsthatmakeinterspecificscentcommunicationlookliketheruleratherthantheexception.Keepingtrackofcompetitorsbyexchangingscentmarksmakesecologicalsense;anyindividualismorelikelytoencounterscentmarksfromanimalsofotherspeciesthanfromitsown,mostcarnivorescompeteforfoodwithmostoftheothers,andtheconsequencesofinterspecificinterferencecompetitionaredireandstronglyasymmetrical.Thisnewcornerofthe

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mammalsemiochemicalfieldpresentsbothopportunitiesandchallenges.Wewillneedtounderstandtheroleofinterspecificsemiochemistryinecologicalinteractionsalongsidetheroleofintraspecificscentsignalsinsocialbehaviourandreproduction.Ifinterspecificscents’mainroleistosignalmerelytherecentpresenceofacompetitortheymaybemoreaccessibletobeingartificiallyrecreatedthanareintraspecificscentswiththeirsignalsofmultiplereproductiveandsocialparametersandindividualsignatures.Ontheotherhand,ifdifferentspeciesareovermarkingoneanother,howdoweevercollectacleanscentmarkfromthefield?ThematicSessionIV:Inter-specificolfactorycues(Chair:PeterApps)Talk:EarlyOlfactoryExperienceAffectsPerceptionofPredatorOdorsintheHouseMouse

V.V.Voznessenskaya,I.G.Kvasha,T.K.Laktionova,A.B.Klinov&M.A.Klyuchnikova

A.N.SevertsovInstituteofEcology&Evolution,Moscow,RussianFederation

UniqueaminoacidL-felinineanditsvolatilederivativesmaybeusedbythehousemousetorecognizepotentialpredators,theirphysiologicalstatusandmayaffectreproductiveoutputinmice.Aimofourstudywastoexaminewhetherearlyolfactoryexperience(EOE)ofmicewithcatchemosignalsmayaffectsensitivitytotargetodorslaterinadulthood,modulatebehavioralorneuroendocrineresponsesandwhetherthesechangesinsensitivitycorrelatedwithneuralactivationinolfactorybulbs.OlfactorythresholdstocaturineandL-felinineweremeasuredwithanautomatedolfactometer(Knosys,USA).FecalglucocorticoidmetabolitesandplasmacorticosteroneweremonitoredusinganELISAtechnique.Behavioralpatternswereanalyzedusinganopenfieldparadigm(2differentmodifications).WeperformedimmunohistochemicalstudiestoidentifyneuralsubstrateinvolvedinreceptionandanalysisofL-felinineandderivatives.Exposuresofmicetocatchemosignals(urineorL-felinine)significantlyloweredtheolfactorythresholds(n=10,p<0.05;n=10,p<0.01).EOEwithcaturine/felininealsodecreased(n=22,p<0.01)patternsofpassive-avoidancebehaviortocatodorsandsignificantlyelevatedinvestigatoryactivity(n=22,p<0.01).Atthesametimecorticosteroneresponsetocaturine/L-felininestayedunchanged(n=10,p<0.01)indicatingtheinnatenatureoftheresponse.Werecordedspecificpatternofactivationinaccessoryolfactorybulb(AOB)inresponsetostimulationwithL-felinine(0.05%).EOEwithL-felinine(0.05%)causedsignificantincreaseinnumberofFos-positivecellsinAOBinresponsetostimulationwithL-felinine(n=8,p<0.01)aswellaswerecordedanincreaseofactivatedarea(n=8,p<0.001).Talk:Ontherepellentpropertiesofmouseurineodourtosmallwildlivingrodents

E.V.Kotenkova,I.A.Zhigarev,V.V.Alpatov,A.O.Shchukin,V.A.Babikov&Yu.A.Bazhenov

SevertsovInstituteofEcologyandEvolutionRAS,Moscow,RussianFederation

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Synanthropicmicespeciesarecommonpeskyorganismsinhumanenvironments.Whiletheirpresenceisubiquitoushomeownersoftencomplainabouttheirunique,persistentandpotentodourwhichisderivedmainlyfromtheirurine.Evolutionarilyitisnotfavorable.Itsuggeststhatthefeatureisnotneutralandlikelyshouldbeusefulformiceforotherreasons.Wehypothesizedthatsuchodourcouldhavebeendevelopedduringevolutionasadaptationdirectedtoprotectionofbuildings,specialecologicalnichecreatedbyhumans,fromotherrodentspecies.AdultindividualsofoutdoorspeciesClethrionomysglareolusandfacultativesynanthropicspeciesMicrotuslevisavoidedthearmofY-mazescentedbyM.musculusurineincomparisonwitharmwithwaterintwo-choicetests.TheyalsopreferarmscentedbyurineofrabbitoroutdoormouseMusspicilegusincomparisonwitharmscentedbyM.musculusurine.InfieldexperimentsyoungandjuvenileCl.glareolusselectivelychoosetoentertrapswithwaterincomparisonwithtrapsscentedbyurineofM.musculus.Theseresultsindicaterepellentpropertiesofmouseurineodourtootherrodents.Wespeculatethatolfactorycuesofhousemicefamilycouldpreventinhabitationofbuildingsbyotherspeciesofrodents.Potentialroleofsulfurcompoundsinmouseurineasrepellentsisdiscussed.SupportedbyRussianScienceFoundation(project№16-14-10269).PosterTalks3:Inter-specificCues&OtherContributionsPOSTER9:EffectofDomesticCatChemicalSignalsonRegulationofEstrousCyclesintheHouseMouseDependsonPresentationRate

T.K.Laktionova,I.Kvasha&V.V.Voznessenskaya

A.N.SevertsovInstituteofEcology&Evolution,Moscow,RussianFederation

L-felinineisauniqueaminoacidfoundintheurineofdomesticcatFeliscatus.WeexaminedtheinfluenceofL-felinineonregulationofestrouscyclesinmice.Inordertomonitorestrouscyclephase(ECP)wevalidatednoninvasiveestradiolassessmenttechnique.TocontrolECPvaginalsmearsweretakenandfecalestradiollevelsweredeterminedforeachfemaleondailybasis.Weused4groupsofmice(n=38)atageof3months:(1)continuousactionofL-felinine(0.05%;50µl)during12days;(2)applicationofL-felinine(0.05%;50µl)atregularintervalsfordurationoftwohoursondailybasis;(3)spontaneousexposurestoL-felinine(0.05%;50µl)butthetotaltimeofexposureduring12dayswasalso24hours;(4)control(exposuretotapwater).Estradiolbaselinewascalculatedindividuallyforeachanimal;concentrationsabovethebaselinewereconsideredasabeginningoflutealphase.ThedataobtainedindicatethatL-felininemayaffectthelengthofestrouscycleinmice.Thenumberofovulationsinthegroup1significantlyincreased(p=0.00498,n=10).Itappearstobethatfemaleshabituatedtothecontinuousactionofthecompound.Atthesametimeweobserveddeclineinanumberofcyclingfemalesinthegroup2(p=0.0233,n=9).Forthegroup3weobservedonlyatendencytodecreaseinanumberofcyclingfemales(p=0.0578,n=9).DifferentmodesofexposuretoL-felinineproduceddifferenteffectonestrouscyclesin

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mice.SupportedbyRFBR16-34-00872POSTER10:Preliminaryresultsinferretolfactorycues(Mustelaputoriusfuro)asapredatorstimulusforthehousemice(Musmusculus)

P.C.Grau,J.Leclerq,E.Teruel,C.Lafont-Lecuelle&P.Pageat

ResearchInstituteinSemiochemistryandAppliedEthologyApt,France

Thehousemice(Musmusculus)asothersmallrodentsareinthebaseofvertebratepredator’strophiccascades.Theyarethemostwidespreadmammalonearthafterhumans,whichalongwithitsfastsexualcycleandprolificitymeanslargepopulationsandabasicsourceofnutrientsforawidespectrumofpredators.Asmacrosmaticanimals,miceuseolfactionasaprimarytooltoavoidpredators,howeverlittleisknownaboutthepredatorolfactorycuesandbehavioralreactionslinkedtothesestimuli.Withthisstudyweperformedapreliminaryapproachtomammalianandreptilianolfactorypredatorycuesofthehousemice.Forthisporpoisewecarriedoutachoicetest,wherewemeasuredduring10minutesthetotaldurationthatmiceremainedinthenearbyareaorfarendareafromthepredatorystimulus,micehadnophysicalaccesstothestimulus,andbothpartswereidentical.Ourresultsshowedthatmicesignificantlyavoidedferretolfactorystimulifromfurandfaeces.Theseresultsareinlinewitharecentstudythatshowedavoidanceofhamsterstoferreturinedependingontheferret’sdiet(Apfelbach,Soini,Vasilieva,&Novotny,2015).Howeverfurtherresearchshoulddelveinferret’solfactorycuesandsemiochemicalsasasignificantrodentpredator.POSTER11:Measuringlactationinvestmentamongcommunallynursingfemalehousemice

A.Davidson1,J.P.Green1,C.DeMatosFerrazFranco2,V.Lee2,PStockley1,R.J.Beynon2&J.L.Hurst1

1MammalianBehaviourandEvolutionGroup,InstituteofIntegrativeBiology,UniversityofLiverpool,LeahurstCampus,Neston,L647TE,UnitedKingdom2CentreforProteomeResearch,InstituteofIntegrativeBiology,UniversityofLiverpool,CrownStreet,Liverpool,L697ZB,UnitedKingdom

Wildfemalehousemice(Musmusculusdomesticus)nestsociallyandoftenrearoffspringcommunallywithchosennestpartners.Cooperationamongcommunallynursingfemalesyieldsimportantfitnessbenefits,includingalargernumberofoffspringweanedandincreasedweightofpups.Ithasalsobeenshownthatfemalesprefertonestandrearoffspringcommunallywithsisters.Althoughfemalesappeartocareforoffspringincommunalnestsindiscriminately,littleisknownaboutthelactationinvestmentmadebyeachmotherinherownoranestpartner’soffspringbecauseofthedifficultyofmeasuringthis.Toovercomethischallenge,wedesignedanexperimentinwhichtwocommunallynursingfemalesfedonseparatedietssupplementedwithanaminoaciddifferentially

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labelledwithstableisotopes,usingcustom-designedfeeders.Astheisotopelabelstransfertotheoffspringviaeachfemale’smilk,theratioofthesetwolabelsallowedustoquantifythepreciseamountoflactationinvestmentfromeachfemaleineachindividualpupinthecommunalnest.Weestablishedcommunallynestingpairsoffemalesthatwereeitherfamiliarlittermatesistersorunrelatedandunfamiliaruntiltheywerepairedasadults.Thisisallowingustoaddresswhetherfemalesinvestmoreintheirownoffspringthaninthoseofanotherfemale;whetherlactationinvestmentismoreegalitarianbetweentwosisterscomparedtounrelatedfemales;andwhetherfemalesthatgivebirthsecond(i.e.haveyoungerpupsinthecommunallitter)investmoreintheoffspringofbothlitters.POSTER12:Latrineusepatternsofraccoondogs(Nyctereutesprocyonoides)inanurbanareaofTokyo

M.Tsunoda1,3,Y.Kaneko1,T.Sako2,R.Koizumi1,K.Iwasaki1,I.Mitsuhashi1,M.U.Saito1,C.Newman3,D.W.Macdonald3&C.D.Buesching3

1CarnivoreEcologyandConservationResearchGroup,TokyoUniversityofAgricultureandTechnology,Japan2ImperialHouseholdAgency,Japan;3WildlifeConservationResearchUnit,UniversityofOxford,UK

Althoughurbanizationisoneoftheleadingthreatstowildlifeconservation,somespecieshaveadaptedtolivingalongsidehumans.Hereweusedapopulationofwildraccoondogs(Nyctereutesprocyonoides)intheAkasakaImperialGroundsincentralTokyoasamodeltoinvestigatehowlatrineusingcarnivorescanmaintaintheirsocio-spatialorganisationunderhumandisturbance.Between2012-2014weemployed4-11cameratrapspositionedatatotalof12differentlatrineslocatedindifferenthabitattypesandsubjectedtovaryinglevelsofhumandisturbance,toanalysethetemporalpatternsofatotalof2459latrinevisits,ofwhich878includeddefecationevents.Overall,latrinesdisturbedbyhumansreturnedlessdaytimevisits(withandwithoutdefecationevents)thanundisturbedonesandactivityshiftedfromday-tonight-time.Latrineuse(i.e.,visitswithandwithoutdefecationevents)increasedtowardswinter,coincidingwithdispersal,andshowedaseasonalshiftfromdiurnaltonocturnalusagepatterns.GLMMresultsalsoconfirmedthatraccoondogsdefecatedatopenlatrinesmorefrequentlythanatshelteredones.Raccoondogslikelyavoidtheperceivedpredationriskfromhumandisturbancebyshiftingtheirvulnerableactivity(i.e.,defecating)tominimisedisturbance,wheretheinherentlongevityofolfactorysignalsallowsparticularlymacrosmaticcarnivorestomaintaintheirsocio-spatialorganisationthroughlatrine-centredolfactorycommunication.Wepositthatthesebehaviouralmodificationsmayallowraccoondogstomaintaintheirsocio-spatialpopulationstructureeveninurbanisedareasthatareaffectedbyhumandisturbance.

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Poster13:Tastepotentiatedcolourbiases:analternativemethodforassessingtastedetectionthresholdsinbirds

H.M.Rowland

DepartmentofZoology,UniversityofCambridge,UK;MaxPlanckInstituteforChemicalEcology,Germany

Understandinghowbirdpredatorsassessthechemicalcontentofpreyisrelevanttoourunderstandingoftheadaptivesignificanceofpreywithwarningcolours;itisalsorelevanttothestudyofaviantasteperception.Birdsareoftenassumedtohaveapoorsenseoftastebecauseofthereducednumberoftastebudscomparedtoothervertebrates(~300inbirdscomparedto9000inhumans).Birdsalsohaveareducedtastereceptorgenerepertoiresizecomparedtoothervertebrates.Forexample,domesticchickenshaveonlythreebittertastereceptorgenesintheirgenome(ggTas2r1,ggTas2r2andggTas2r7I)comparedwith51receptorgenesinfrogs.Contrarytotheassumptionthatbirdshaveapoorsenseoftaste,Ishowthatsomechemicalsseemtobeintrinsicallyaversivetobirds,andthatbirdsshowconcentration-dependentresponsestobitter-tastingchemicals.Iusedacolour-biasparadigmtoexaminetheresponsesofchickenstodifferentconcentrationsofquinine,denatoniumbenzoate,quassiaandcaffeine.Thisassessmentofunlearnedresponsestonaturallyoccurringdefensivechemicalsisanalternativemethodforassessingdetectionthresholdstotheforced-choicesolution-consumptionmethod.

AMCoffeeBreak

ThematicSessionV:ScentofDisease(Chair:StefanieGervasi)Talk:LPS-inducedimmunesystemstimulationalterurinaryvolatilesandbehavioringrowingpigs

S.G.Devaraj1,A.Rakhshandeh2,J.McGlone1

1LaboratoryofAnimalBehavior,PhysiologyandWelfare,AnimalandFoodSciencesDepartment,TexasTechUniversity,Lubbock,TX79409;2AnimalandFoodSciencesDepartment,TexasTechUniversity,Lubbock,TX79409

Volatilecompoundsemanatingfrombodysecretionsmayactasanindicatorofthehealthstatusofanimalstootheranimals.Wesoughttoidentifyurinaryvolatilesthatmightactasbiomarkersindicativeofhealthstatusofgrowingpigs.Immunesystemstimulation(ISS)wasinducedinpigs(N=5)byi.m.injectionofbacteriallipopolysaccharide(LPS;25µg/kgBW).Urinesampleswerecollectedbefore(baseline)andafterLPStreatment.TheurinaryheadspacevolatilecompoundswereidentifiedusingSolid-phasemicroextraction(SPME)coupledwithGC-MS.Pigeye/bodytemperaturesweremeasuredusinginfraredimagingtechnique,andbehaviors(i.e.standing,feeding,drinking,moving,lying,sittingandactive)werevideo-recordedandquantified24hbeforeandafterISS.DatawereanalyzedusingmixedmodelANOVAexaminingperiodsbeforeandafterLPSadministrationwithpigsservingastheirowncontrol.Pigeye/bodytemperatureincreasedpost-ISS(at6h,P=0.03)

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andcorrelatednegativelywithactivebehaviors(r=-0.64,P=0.048).Relativetopre-ISS,pigsspentlesstimestanding(P=0.018),moving(P=0.061)andactive(P<0.0001)duringpost-ISSperiod.SPME-GC-MSrevealed3.6-and5.3-foldincreasesin4-Ethylphenol(P=0.005)and3-Methylphenol(P<0.0001),respectively,inpost-ISSurinecomparedwithurinecollectedduringthepre-ISSperiod.InISSpigs,behavioralactivitywasreduced,andexcretionofurinary3-Methylphenoland4-Ethylphenolwereincreased(P<0.01).Changesinbehaviorandvolatilemoleculesmaybeusedasanon-invasivemethodofdetectionoffever,illnessandhealthinfarmandpossiblyotheranimals.Talk:Sharinganenvironmentwithsickconspecificsalterstheodorsofhealthymice

S.S.Gervasi1,B.A.Kimball2,M.Opiekun1,T.Martin1&G.K.Beauchamp1

¹MonellChemicalSensesCenter;²USDA-APHIS-WS-NWRC

Chemicalsignalsconveyvitalinformationaboutenvironmentalrisk.Itiswellknownthatpathogeninfectionaltersbodyodors.“Sicknesssignals”canmediatebehavioralavoidanceofinfectedanimalsbyhealthyconspecificsandaccumulatingevidencealsosuggeststhatexposuretosicknesssignalsactivatesphysiologicalresponsesofsignalreceivers.Sicknesssignalscouldthushavecascadingeffectsontransmissionriskbyinfluencinghostcontactratesandindividualsusceptibility.Weusedanexperimentalapproachtoexamineeffectsofexposuretoodorsofsick,butnon-infectiousindividualsontheodorprofilesofhealthyconspecifics.Weemployedtwotypesofhousingsetups,allowingustodirectlycompareresponsesofpairsoffreelyinteractingorphysicallyseparatedmice.InaY-mazebehavioraltrainingparadigm,biosensormicediscriminatedbetweenodorsofhealthyandsickmice.Unrewardedgeneralizationtrialsconfirmedthatbiosensorsalsodiscriminatedbetweenodorsofhealthymicehousedwithhealthypartnersandhealthymicehousedwithsickpartners.Wefollowedbehavioraldiscriminationassayswithchemicalheadspaceanalysesemployinggaschromatrography/massspectrometry.Chemometricssupportedpatternsobservedinbioassays;thecompositionofodorsofhealthymicehousedwithsickconspecificswasstatisticallydistinguishablefromodorshealthymicehousedwithhealthyconspecificsandmorecloselyresembledodorprofilesofsickanimals.Thus,exposuretosickindividuals,evenintheabsenceoftransmission,couldimpacthowanimalsareperceivedandbehave.Plasticityinproductionofandresponsestosicknesssignalscouldhavecomplexeffectsondiseasedynamicsbymodifyingsocialnetworks.Talk:Diseasedetectioninhumanolfaction

M.J.OlssonDepartmentofClinicalNeuroscience,KarolinkaInstitutet,Stockholm,Sweden.

Contagious diseases have been a fatal threat to people throughout evolution. Recentresearchsuggeststhatbehavioralavoidanceofsickindividualsisthefirst,andprobablymost

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cost effective, line of defense against infection (Schaller, 2011). Indeed, statisticalmodelsshow substantial disease containing effects from small adaptations in patterns of inter-individual contact (Cole, 2006). In addition, behavior can be seen as a vital partcomplementing andeven regulating the classical immune system. This behavioral defenseanditsconsequencesarepoorlyunderstoodandsurprisinglyfewstudiesexist.Wehaveinaseries of studies examined the olfactory cues by which we detect disease; the neuralmechanisms underlying disease avoidance; and how olfactory disease detection preparesthe body for an attack together with classic immunity. In doing this we have utilized anexperimental sickness model involving the induction of innate inflammation with anendotoxin(lipopolysaccharide)injectioninotherwisehealthyparticipants.Resultsshowthatafterafewhoursofsystemicinflammationwesmellmoreaversivefromtheskin,theurinechangesincharacterandshowheightenedconcentrationsofPyrrole.Resultsalsoshowthatdisgusting odors (believed to work as olfactory disease cues) increase levels of aninflammatorymarker (tumor necrosis factor alpha) in the saliva. Altogether these resultssupporttheimportanceofolfactioninbehavioralimmunity.

Talk:Thesmellofweakness:costsandconsequencesofinjuryforsocialodorcuesinring-tailedlemurs

R.Harris1&C.M.Drea1,2

1DepartmentofEvolutionaryAnthropology,DukeUniversity,DurhamNC,USA2DepartmentofBiology,DukeUniversity,DurhamNC,USA

Animalodorantscontaininformationaboutstableandtransientaspectsofthesignaler’sconditionsalienttoconspecifics.Thus,condition-dependentodorscanmediatesocialinteraction,groupdynamics,andsuccessfulreproductioninvertebrates,includingprimates.Scentsignalsarepresumablyenergeticallyexpensivetomaintain,butevidenceofproductioncostsderivesmainlyfromexperimentsontheeffectsofparasitismandinfectioninlaboratoryrodents.Weusedanalyticalchemistryandbioassaystoexaminetheeffectsofconspecific-inducedinjuryonolfactorycuesinanaggressive,sociallycomplexstrepsirrhine,thering-tailedlemur(Lemurcatta).Someinjuredanimalsreceivedantibiotics,sowetestedboththe(1)‘costlysignal’and(2)fermentationhypotheses–thelatterpositingthatsymbioticbacteriainhabitingscentglandssynthesizevolatilesusedbythehost.Weanalyzed111genitalglandsecretionscollectedbefore,during,andafter33injuriesinvolving12malesand10females.Normal,‘healthy’glandularsecretionsweresignificantlydampenedduringinjury,regardlessofantibiotictreatment;secretioncompositionwasdifferentiallyalteredbyinjuryversusmedication.Inbioassays,malelemursdirectedsignificantlymorecompetitivecountermarking’behaviortowardsodorsfrominjuredthanhealthyconspecifics,potentiallyrelevanttosame-sexcompetitionanddominanceinteractions.Wesuggest(1)acosttoodorproductionbornebythesignallerand(2)aroleforfermentativebacteriainsignal

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production.Toourknowledge,thisisthefirstevidencethatmammalianolfactorysignalsareperturbedbyinjury,independentofinfectionordisease.

LunchBreak

ThematicSessionVI:Semio-chemicalMethodologiesandAnalyses(ChairCDBuesching)

Talk:ComplexityofScentMarkinginRats:BehaviouralCharacterisationK.C.Pounder1,G.Gómez-Baena2,J.O.Halstead1,M.Prescott2,R.J.Beynon2&J.L.Hurst1

1MammalianBehaviourandEvolutionGroup,InstituteofIntegrativeBiology,UniversityofLiverpool,LeahurstCampus,Neston,L647TE,UnitedKingdom;2CentreforProteomeResearch,InstituteofIntegrativeBiology,UniversityofLiverpool,CrownStreet,L697ZB,Liverpool,UnitedKingdom

Manyrodentsusechemicalsignalstocommunicate,depositingimportantinformationintheenvironmentintheformofscentmarks.Thesescentsplayafundamentalroleinadvertisinginformationtoconspecificssuchasidentity,sex,age,dominance,reproductivereceptivenessandhealthstatus.Norwayrats(Rattusnorvegicus)areahighlysociablerodentspecies,oftenlivinginlargegroupscompetingforresources,territoryandmatingopportunities,andthusrelyheavilyonchemicalsignalstomaintainsocialstructures.Ratsscentmarkbyloweringtheiranogenitalregiontocontactobjectsintheirenvironment,oftenleavingvisibledepositsofurineand/orsebaceoussecretions.However,relativelylittleisknownaboutthecompositionofscentmarks.Molecularanalysesofmaleandfemalemarksdemonstratethatdepositsarenotsimplyderivedfromurine.Thepreputialandprostateglandsinmales,andtheclitoralglandsinfemales,contributetotheproteincomposition.Weexaminedscentmarkingbehaviourinresponsetopresenceorabsenceofstimuli,aswellasthebehaviouralresponsestonaturallydepositedscentmarksandcomponentsfromdifferentsources,todeterminetheirpotentialroleinintraspecificcommunication.Weshowthatratsarehighlysensitivetotheodourcuesfromscentmarksandcomponentsfromdifferentglands,andeachelicitsex-specificresponses.Maleratsmayalso‘tune’thecontentsoftheirscentmarksaccordingtocontext.Thesefindingsprovidenovelinsightsintothecomplexityofratscentsandtheroleofdifferentscentcomponentsonthenaturalbehaviouralresponsesofmalesandfemales.Talk:PTR-TOF-MSasatoolforstudyingvertebratechemicalecology:acasestudyontheuseinkairomonedeterminationinapredator-preycontext

C.VanMoorleghem1,M.Portillo-Estrada2,Y.Lambreghts1,I.A.Janssens2&R.VanDamme1

1LaboratoryforFunctionalMorphology,DepartmentofBiology,UniversityofAntwerp,Universiteitsplein1,B-2610Wilrijk,Belgium;2CentreofExcellencePLECO(PlantandVegetationEcology),DepartmentofBiology,UniversityofAntwerp,Universiteitsplein1,B-2610Wilrijk,Belgium

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Inthewakeofimprovementsinchemo-analyticalmethods,thefieldofchemicalecologyisrapidlyadvancing.However,particularlyinvertebrates,thetruenatureofinformativecompoundsinspecificcontextsremainslargelyunexplored.Here,wevalidatetheimplementationofarecentlydevelopedtoolusedtodetermineVolatileOrganicCompounds(VOCs),toimprovetheidentificationofsemiochemicalsinawidevarietyofanimalinteractions.TheexistingtechniqueiscalledProtonTransferReactionTime-of-FlightMassSpectrometry(PTR-TOF-MS).Theaccompanyingprocedurehastheadvantageofnotrequiringsamplepreparationwhich,togetherwiththedevice’shighaccuracyandlowdetectionlimit,enablesustomeasureeventhemostvolatilemoleculesthatoftenarelostwithothertechniques.Inthisstudy,weapplythischemo-analyticalmethodfordeterminingcandidatekairomonesusedbyviviparouslizards(Zootocavivipara)todetectandavoidsaurophagoussnakes,suchastheviperViperaberus.Firstly,behaviouralobservationsindicatedthatlizardscanuseairbornechemicalsforpredatoryriskassessment.Subsequently,withPTR-TOF-MS,aVOCprofilewasmadeofViperaberusscentandseveralchemicalcompoundswereputforwardascandidateinformationcarriersonpredatoryrisk.Furthermore,theeffectofodourfade-outonlizardbehaviourandchemicalcompositionwasinvestigated.OurresultsshowthatPTR-TOF-MScouldcomplementmorewidelyusedmethodsinresolvingimportantunansweredquestionsinvertebratechemicalecology.Talk:Non-invasivesamplingofbodyodourtoinvestigatetheinformationcontentofmeerkatscent

B.M.Weiß1,2,A.Marcillo3,M.Kücklich1,2,M.Manser4,R.Holland5,C.Birkemeyer3,A.Widdig1,2,6

1InstituteofBiology,UniversityofLeipzig,Germany;2JuniorResearchGroupofPrimateKinSelection,DepartmentofPrimatology,Max-Planck-InstituteforEvolutionaryAnthropology,Leipzig,Germany;3InstituteofAnalyticalChemistry,UniversityofLeipzig,Germany;4DepartmentofEvolutionaryBiologyandEnvironmentalStudies,UniversityofZurich,Switzerland;5ZooLeipzigGmbH,Leipzig,Germany;6GermanCenterforIntegrativeBiodiversityResearch(iDiv),Leipzig,Germany

Theolfactorycuespresentinananimal'sbodyodourmediateawiderangeofmammaliansocialinteractionsbyencodinginformationaboutindividualattributessuchasidentity,sex,age,orgroupmembership.Thechemicalcompositionofthesecuesprovidesimportantindicationsoftheecologicalandevolutionaryprocessesshapingolfactorycommunication.However,whileolfactorycommunicationisfrequentlyaddressedinbehaviouralstudies,thecompoundsinvolvedinchemicalinteractionsbetweenindividualshaveonlybeensparselyinvestigatedinmostmammaliantaxaandcontexts.Weadjustedandevaluatedtheuseofthermaldesorption(TD)tubes,commonlyusedinplantmetabolomicandenvironmentalstudies,fornon-invasivesamplingofmeerkat(Suricatasuricatta)bodyodour.Adjustmentofsamplingparameters(e.g.distance,volume,flowrate,interruptionofsampling)toincreasefeasibilityforanon-invasiveapplicationyieldedsamplesofadequatequality.TDtubes

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capturedawiderangeofvolatileandsemi-volatileorganiccompoundsincludingthoselikelyoriginatingfromthetargetanimals.Thecompositionofthebodyodourprofilesdifferedbetweengroups,ageclassesandbodyregions,butnotbetweenthesexes.Theseresultscorroboratefindingsofbehaviouralstudiesonmeerkatolfactorycommunicationandondifferencesinbacterialcommunitiesinmeerkatscentsecretions.OurstudyindicatesthatTDtubesaresuitablefornon-invasivesamplingofbodyodour,makingthemanattractivetoolforaddressingawiderangeofquestionsinthelabandthefield.Talk:Themaleeffect:findingawaythroughthejungleofchemicalcues

C.LeDanvic1,M.Dillenbourg1,M.Keller2,L.Schibler3,P.Nagnan-LeMeillour4

1ALLICE,R&DDepartment,UGSFUMR8576USTL/CNRS,VilleneuveD’Ascq,France;2INRA,CentredeTours,Nouzilly,France;3ALLICE,R&DDepartment,Paris,France;4INRA,USCINRAUGSF,VilleneuveD’Ascq,France

Inungulates,theintroductionofasexuallyactivemaleinananoestrusfemaleherdinducesasurgeofLHleadingtoovulation.Thisprocessconventionallyusedbybreedersintheirflock,knownas"maleeffect",allowsthecontrolreproductionandappearstobealsoaninterestingalternativetotheuseofhormone.Maleeffectdependspredominantlyonolfactorystimuligeneratedbythemale(Okamura&Mori,2005;Cohen-Tanoudjietal.,1994).Ifsomepotentiallyinvolvedmoleculeshavepreviouslybeenidentified(Cohen-Tanoudjietal.,1994;Murataetal.,2014),thecompletesignal(thoseinducingovulation)remainstobeclearlycharacterized.Usinganoriginalapproachbasedonthecomparisonofolfactoryprofilesofmalesduringnon-reproductiveandreproductiveperiods,weundertookthecomplete“maleeffect”pheromonescharacterizationinFrenchbreeds.Variousbiologicalsamples(urine,wool,headsecretions)thatcouldberesponsibleformaleeffectwerecollectedandchemicalprofilesweredeterminedbyGC/MSanalysisafterSPMEorsolventextractions.Specificramandbulkolfactoryprofileshavebeenobservedduringtheperiodofmaximalsexualactivitywith1/aglobalincreaseinintensityprofilesand2/theappearanceofnumerousmolecules.Intotal,morethan60(on147)moleculesseason-specific(Fold-change>10)werecharacterized(ethylesters,ketones,…).Wearenowfocusingourinterestonthosemoleculestovalidatetheirbiologicalactivityonfemale.Discussionsarecurrentlyconductedwithourpartners(INRA,CNRS)todefinethebestmodalitiestoassesstheirbiologicalactivity.Thelong-termobjectiveistodevelopadifuserallowingtomimic/potentiatethemaleeffectinbreedingfarms.Talk:Comparisonofdifferentanalyticalmethodsforpre-processinggaschromatographicdatainsemio-chemicalresearch

C.D.Buesching1,H.V.Tinnesand2,C.T.Mueller3&M.J.Noonan41WildlifeConservationResearchUnit,UniversityofOxford,ZoologyDepartment,TheRecanati-KaplanCentre,TubneyHouse,AbingdonRoad,Tubney,Abingdon,OX135QL,UK;

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2FacultyofArtsandSciences,DepartmentofEnvironmentalandHealthStudies,TelemarkUniversityCollege,N-3800BøiTelemark,Norway;3CardiffUniversity;4SmithsonianConservationBiologyInstitute,NationalZoologicalPark,1500RemountRd.,FrontRoyal,VA22630,USA.

Recentadvancesinsensitivityandcost-effectivenessofGC-MStechniqueshaveresultedinadrasticincreaseofsemio-chemistryresearch.Problematically,however,GC-MSdatadonotcarryabsolute,butratherrelativeinformation,andthusevensmalldifferencesinsamplevolume,concentration,and/orlabprocedures,incombinationwithbackgroundnoise,caninfluencetheperceivedabundanceofcompounds.Becauseofthis,pre-processingmustbecarriedoutbeforedatacanbecomparedstatistically.Currently,researchersuseawiderangeofpre-processingtechniques,oftenunawareofsomeoftheirintrinsicpropertiesandlimitations.Here,weuseacombinationof20,000simulatedprofiles,andempiricalGC-MSdatafrom15samplesofbadger(Melesmeles)subcaudalglandsecretion,totesttheperformanceofthemostcommonlyusednormalisationmethods(TotalSumNormalisation‘TSN’;ProbabilisticQuotientNormalisation‘PQN’;InternalStandardNormalisation‘ISN’;ExternalStandardNormalisation‘ESN’;andacompositionaldataapproach‘CODA’).Wedemonstratethateachmethodimpartsuniquepropertiestothedata,andanyecologicalinferenceisthuslikelytobeinfluencedbythenormalisationtechniqueemployed.Calibratingagainstanexternalorinternalstandardprovidedgoodresultswhendifferenceswerepronounced,butwerelessreliableformoresubtlydifferentiatedgroups.ThereliabilityofTSNandCODAapproaches,however,werehighlydependentonthestructureofthedata,andpronetogeneratingerroneousconclusions.ThelowfalsepositiverateofthePQNapproachmadeitthemostgenerallyapplicablenormalisationmethod,whereasthesuggestionofutilisingablanketapproachoffirstapplyingmultipledifferentnormalisationmethodsandthenidentifyingresultsthatareconsistentacrosseachisunlikelytobeeffectiveforalldatasets.Talk:SamplingandanalysisofVOCs&SVOCs–Whatarethebesttoolsforthejob?

MassimoSantoro,IlariaFerrante,HannahCalder

MarkesInternationalLtd.,GwaunElaiMedi-ScienceCampus,Llantrisant,UK

Thermaldesorption(TD),inconjunctionwithgaschromatography(GC),isapowerfulandversatiletechniquethatenablesthedetectionandquantitationoftraceorganicchemicalsfromambientair,workplaceenvironmentsaswellasthosereleasedfrommaterials.Morerecentlythiswellusedtechniquehasbeenadoptedbythefieldsofchemicalecologyandbiologicalmonitoringandoffersfascinatinginsightsintothecomplexinteractionsinvolvingplants,microorganismsandanimalsaswellasthedetectionofdiseasemarkersandodouranalysisofbreath.Thispresentationwillfocusonthevarioussamplingtechniquesthatfacilitatetheuseforthistechniqueacrossawiderangeofapplicationsanddiscussthe

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benefitsoftheanalyticalsystemfordetectionandquantitationofVolatileandSemi-volatileorganiccompounds.

PMCoffeeBreak

Opensession4Talk:Temporalchemicalprofilingofurinevolatilecompoundsofdomesticcatsandtheirolfactorydiscriminationabilities

C.Suzuki,M.Maita,T.Miyazaki,T.Yamashita&M.Miyazaki

DepartmentofBiologicalChemistryandFoodSciences,FacultyofAgriculture,IwateUniversity,Japan

Territorialanimalsdeposittheirurineandfecesontheirterritories.Theseexcretionscontainthousandsofmetabolitessomeofwhichmaybeusedasscentsignalsforspecies,sex,age,andindividualrecognitions.Previousstudieshavesuggestedthatcompoundsidentifiedspecies-,sex-,andage-specificallyandvariedamongindividualsareimportantforscentcommunication.Inthesestudies,freshexcretionswereusedfortheirchemicalprofilingandbehaviorbioassays.Howeveranimalsusuallyencounterexcretionsafteragingperiodsofseveralhoursanddaysintheirterritories.Therefore,itisnecessarytoanalyzenotonlyfreshexcretionsbutalsoagingexcretionsinanimals.Inthepresentstudy,weexaminedtemporalchangesofchemicalprofilesofurinaryvolatilecompoundsofdomesticcats,andtheirolfactorydiscriminationabilitiestowardthesevolatiles.Cottonswereinfiltratedwithmalecaturine,andthentheheadspacegasemittedfromthecottonsbeforeandafterair-dryingwereanalyzedintwo-dimensionalgaschromatography(GC×GC)-massspectrometry(MS).GCxGC-MSdetectedovertwothousandsofvolatilesinbothheadspacegasemittedfromfreshandagingurine.Chemicalprofilesoftheheadspacegasdifferedmarkedlybetweenfreshandagingurine.Therewasatendencythatcontentsofcompoundswithlow-boilingpointdecreasedinagingurine,whilecompoundswithmiddle-boilingpointincreased.Inbehavioralassays,catswhichhabituatedtofreshurineshoweddishabituationtoagingurine.Theseresultsindicateagingurineemittedmorecompoundswithmiddle-boilingpointthanfreshurine,andcatscandistinguishdifferencesofchemicalprofilesbetweenfreshandagingurinebyolfaction.Talk:Identificationofkeycompoundsfortheindividualrecognitioninthedomesticcats

M.Maita1,C.Suzuki1,J.Caspers2,T.Yamashita1,S.Schulz2&M.Miyazaki1

1DepartmentofBiologicalChemistryandFoodSciences,IwateUniversity,Iwate,Japan;2DepartmentofLifeScience,TechnicalUniversityofBraunschweig,Braunschweig,Germany

Domesticcatsspraytheirurineontoaverticalsurfaceforscentmarking.Theysniffurinedepositedbyotherindividualsofintraspeciescarefully,andthenexhibittheflehmenresponse;thecatsraisetheirheadandholdtheirmouthpartiallyopenforafewseconds.Ourpreviousstudiesdemonstratedthatvolatilefelininederivativeswerestrongcandidatesofscentsignalsforspecies,sex,andageinformationincats.However,littleisknownwhich

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compoundsareinvolvedinindividualrecognitionincats.Theaimofthisstudywastoidentifykeycompoundsforindividualrecognitionincats.Itisknownthattheflehmenresponseisobservedinmalesthatsniffestrousfemaleurineincats.Inaddition,wefoundthatcatsexhibitedtheflehmenresponsetoothercaturineofsamesex,butnotownurine.Wepurifiedkeycompoundsfromcaturine,usingsniffingtimeandtheflehmenresponseasindicatorsofindividualrecognition.Ouranalysesindicatedthatcatssniffedurinarycompoundscontainingofafunctionalgroupoflipidswithconsiderableinterestsandshowedtheflehmenresponse.InGC-MSanalyses,urinarycontentsofthesecompoundswerevariedamongindividuals.Habituation-dishabituationtestsshowedthatcatsdistinguishedthedifferenceofurinarycontents.Furthermore,chemicalprofilesofthesecompoundswereconservedbetweenfreshandagingurineineachcat.Theseresultsindicatethatthelipidsarekeycompoundsforindividualrecognitionincats.

Friday01September2017

Plenary:Scentwars:unravellingthecomplexityofcompetitivescentsignalling

J.L.Hurst

MammalianBehaviourandEvolutionGroup,InstituteofIntegrativeBiology,UniversityofLiverpool,LeahurstCampus,NestonCH647TE,UK

Competitivescentmarkingisthemostovertformofscentsignallingseeninmammals,involvingabattlefoughtbetweenindividualsorsocialgroupsthatisplayedoutatbothbehaviouralandmolecularlevels.Competition,particularlyinthecontextofreproductiveopportunity,oftenleadstoadramaticincreaseininvestmentinspecificmolecularcomponentsofscentsaswellasinbehaviouraldeployment.However,anindividual’sabilitytoproduceanddeploythesesignalsisregulatedthroughcompetitiveaggression,leadingtohonestsignallingofcompetitiveability.Studiesinrodents–mostparticularlyinhousemice–haveidentifiedmultiplecomponentsthatcontributetothesedynamicsignals,potentiallyprovidingextensiveinformationabouttheownertocompetitorsandtopotentialmates.Thisincludesseveralandrogen-dependentvolatilepheromonestogetherwithMUPs(majorurinaryproteins),whichareamultigenefamilyofinvolatileurinarylipocalinproteinsthatbindandcontrolthereleaseofvolatilesfromdepositedscentmarks.Onemale-specificMUP,darcin,playsakeyroleininducingfemalemicetorapidlylearnattractiontotheodourandlocationof‘winning’malesthatcountermarktheircompetitors’scents.Mostinvestment,though,isinanothersetofMUPsthatprovideadistinctiveandsurprisinglystablesignatureofowneridentityatseverallevels.ThisMUPsignaturefurtherdeterminesthevolatilesignaturethatfemaleslearnfrommalescentmarks.IncreasingtherateofscentdepositionandamountofMUPinvestmentthusincreasesthestrengthofanindividual’sscentinthefaceofcompetingsignals,providingacombinedsignalofidentityandcompetitivestatus.

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ThematicSessionVII:Olfactoryadvertisement(ChairJaneHurst)Talk:Urinaryvolatilesandbioindicatorsrelatedtoreproductivestatusingiantpanda(Ailuropodamelanoleuca)

M.Dehnhard1&J.Brown2

1LeibnizInstituteforZooandWildlifeResearch,Berlin,Germany(dehnhard@izw-berlin.de);2CenterforSpeciesSurvival,SmithsonianConservationBiologyInstitute,FrontRoyal,USA

Inex-situpandabreeding,urinarysteroidsserveasestrousindicatorsandtotimeAI.Followingaperiodofadelayedimplantation,femalepandasgenerallyexperiencepseudopregnancy.However,ifatruepregnancyoccurs,aplacentaisformedthatmayaffectthecompositionofurinarycompounds.Weaimedto:1)analyseurinaryvolatilesthatadsorbon1cmpiecesofpolydimethyl-siloxane(PDMS);2)detectestrousrelatedsubstances;and3)findcompoundsthatmightbeapplicabletodifferentiatetruepregnancyfrompseudopregnancy.AnAgilentGCMSwasusedfortheanalysesofPDMSadsorbedsubstancesusing1mlofurine.CompoundprofilingwascarriedoutusingthefreelyavailableXCMSsoftwareathttp://metlin.scripps.edu.AnalysesofPDMSadsorbedurinaryvolatilesconfirmedearlierresultsofanabruptincreaseinfattyacidsfrombasaltopeaklevels6dayspriortoovulation.However,fattyacidcompositionappearstovarybetweenconsecutiveyearsandbetweenindividuals.Inaddition,substancessuchasD-limoneneand2-ethyl-1,4-dimethyl-benzeneweredetectedpeakingaroundestrous.Comparisonofurinarycompoundsbetweenpregnanciesandpseudopregnanciesfromonepandaduringthesecondaryphase(highlevelsofprogestagensfollowingassumedimplantation)revealedseveralsignificantlyelevatedbutunidentifiedsubstancesduringtruepregnancy.Moreover,oneofthosesubstanceswasalsosignificantlyelevatedduringtheprimaryphase(lowlevelsofprogestagensafterconception).However,nosubstancewasfoundexclusivelyduringtruepregnancy.Asresultswereobtainedfromonlytwopandas,moresamplesareneededtoconfirmifapregnancyspecificsubstanceexistsinthisspecies.Talk:Femalesprefercooperativemalesbasedoncuesdifferentfromcooperativebehaviours:evidencefromtheMound-buildingmouse,Musspicilegus

A.Tognetti1,M.Raymond2,G.Ganem2&C.Faurie2

1InstituteforAdvancedStudyinToulouse,21alléedeBrienne,31015Toulouse–France;2InstituteofEvolutionarySciences(ISEM),UniversityofMontpellier,CNRS,IRD,EPHE,placeEugèneBataillon34095Montpellier–France

Theoreticalandempiricalstudiesinhumanssuggestthatcooperativebehaviorsmayactassignalsduringmatechoice.However,cooperationisnotalwaysobservablebypotentialpartnersbeforematechoice.Toaddresswhethercooperativephenotypesarepreferredbasedoncuesdifferentfromcooperativebehaviorsperse,wedesignedanexperimentalparadigmusingwild-bornMound-buildingmouse(Musspicilegus)aspecieswithbiparentalcare.Inthisspecies,femalescannotobservemalecooperativebehaviors:matechoiceoccursinspring,whereasmoundsarecooperativelybuiltinfall.Wefirstassessedvariation

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inmoundbuildinginvestmentandidentifiedhighandlowcooperativemales.Second,wepresentedthesemalestofemalesduringtwo-waychoicetests.Ourresultsindicatethatfemalesweremoreattractedtohighoverlowcooperativemales,evenwhentheydidnotobservethembuild.Thisfindingsuggeststhatfemalematechoiceisinfluencedbycuesofcooperativenessdifferentthancooperativebehaviorsperse.Wespeculatethatsomechemosensorycuesofmatequalityorcooperativenesscouldbeinvolvedinthisdetection.Talk:Sex-steroidexcretioninurineasolfactoryadvertisementinEuropeanbadgers(Melesmeles)inrelationtocirculatingplasmatitres.

N.A.Sugianto1,H.V.2,D.W.Macdonald1&C.D.Buesching1

1WildlifeConservationResearchUnit,DepartmentofZoology,UniversityofOxford,Recanati-KaplanCentre,AbingdonRoad,TubneyHouse,Tubney,OxfordshireOX135QL,UK.2FacultyofArtsandSciences,DepartmentofEnvironmentalandHealthStudies,TelemarkUniversityCollege,N-3800BøiTelemark,NorwayCirculatingsex-steroidlevelsareareliableindicatorofanindividual’sreproductivestatus.Aftermetabolization,sex-steroidsareremovedfromthebloodstreamthroughexcretioninurineviatherenalsystem,orinfaecesviatheliver.Therefore,thesemetabolicwasteproductsmayserveashonestandreliableolfactoryreproductiveadvertisementsignalstoconspecifics.TheEuropeanbadger(Melesmeles)isahighlymacro-osmaticmustelidandusesglandularsecretionsaswellasurineandfaecesforintra-specificcommunication.Here,weanalysed127urinesamplescollectedfrom58maleand69femalebadgersinWythamWoods(Oxfordshire-UK)duringthepost-weaningperiodforfemalesandmainmatingseasoninspring(May/June),duringthereportedsecondarymatingpeakinsummer(August/September),andduringreproductivequiescenceinautumn(November),andusedGC-MStodeterminehowurinarysex-steroidmetabolitesdifferbetweenindividualsofdifferentreproductivestates.GC-MSprofilesvariedwithcirculatingplasmalevelsofoestradiol(measuredwithEIAusingoestradiol-17-HS-BSA-antiserum),oestrone(EIAusingpolyclonal-EC-R522-antiserum)andtestosterone(EIAusinganti-testosterone-R156/7).Asbadgersarereportedtoboth,sniffandlickurinefromconspecifics,weinferthattheylikelyusevolatileaswellasnon-volatilecomponentstodecipherurinaryinformationcontent.Talk:Scent-markingasHonestFitnessAdvertisementinEuropeanBadgers(Melesmeles)

T.Allen,C.D.Buesching,C.Newman&D.W.MacdonaldWildlifeConservationResearchUnit,DepartmentofZoology,UniversityofOxford,Recanati-KaplanCentre,AbingdonRoad,TubneyHouse,Tubney,OxfordshireOX135QL,UK.Scent-markingisconsideredcostlynotonlyduetothephysicalactivityassociatedwithdepositingscentmarkswithadequatefrequencyandofadequatestrength,butalsoduetothephysiologicalproductionofthesecretionsthemselves.Factorsrelatedtohonestfitnessadvertisement–particularlyindividualvariationinsecretioncomposition–duetointrinsic(e.g.age,sex,genetics)andextrinsic(e.g.resourceavailability,populationdensity,climate)factorscanimpacttheperceivedattractivenessofscent-marks.Therefore,scentmarkshavebeensuggestedtofunctionashonest

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fitnessadvertisementsignals.However,littleresearchhasinvestigatedindividualenergeticcoststowardsecretionproduction.Europeanbadgers(Melesmeles)arehighlymacrosmaticmustelids.Likeallcarnivores,theypossesspairedanalglands,buthaveevolvedanadditionaluniquesubcaudalgland.Subcaudalglandsecretionisrichinlong-chainedcarboxylicacidsandencodesforindividualityandgroup-membership,andawealthoffitness-relatedparameters,suchassex,age,reproductiveandsomaticcondition.Herewemeasuredthesizeofthesubcaudalglandopeningasaproxyforscentmarksize,andanalysedtheenergycontentofsubcaudalglandsecretions,usingbombcalorimeteranalyses.Werelateopeningsizeandcalorimetricdifferencestosex,ageandbodycondition,anddiscussthesuitabilityofsubcaudalglandsecretionsashonest(i.e.unfakable)fitnessadvertisementsignals.

AMCoffeeBreak

PosterTalks4:AdvertisementPOSTER14:Changesincompositionofurineduringvariousperiodsofovariancycleinfemaleofdomesticdogs(Canisfamiliaris)

M.Dzięcioł1,A.Szumny2,W.Niżański1,T.Jezierski3,M.Woszczyło1&B.Pieczewska1

1WroclawUniversityofEnvironmentalandLifeSciences,DepartmentofReproductionandClinicofFarmAnimals,50-366Wrocław,PlacGrunwaldzki49,Poland;2WroclawUniversityofEnvironmentalandLifeSciences,DepartmentofChemistry,50-375Wrocław,C.K.Norwida25,Poland;3InstituteofGeneticsandAnimalBreedingofPolishAcademyofSciences,DepartmentofAnimalBehavior,Jastrzębiec,05-552Magdalenka,Poland

Sincemanyreportsunderminesthemeaningofmethylparabenasasexpheromonesindogs,theaimofthisstudywastoevaluatechangesincompositionoftheurineinfemaledog(Canisfamiliaris)duringthedifferentphasesofovariancycle.MorethanhundredofurinesamplescollectedfromthevariousfemalesduringtheparticularphasesofovariancyclewereevaluatedwiththeHS-SPME/GC-MStechnique.Thephaseofthecyclewasconfirmedbyevaluationofclinicalsymptomsaswellasbyvaginalcytologyandhormoneslevelmeasurement.Theresultsshowedsignificantdifferencesbetweensamplescollectedduringparticularphasesofovariancycle.Aboutthirtycompoundswereidentifiedinurinesamples.Severaldifferencesinchemicalprofileofvolatilecompoundsasaffectedbyovariancyclewerenoticed.Unequivocally,thelevelofcarbonylaromaticcompounds,suchasacetophenone(hypnone)andbenzaldehydeincreasedintheoestruscycle.Itwasaccompaniedbyrisingthemethylketones,e.g.butanone,2-pentanone,3-hexanone.Simultaneouslythesulfidecompounds:1-methylotiopropane;1-methylotiobutane,1-methylotipentaeordimethyltrisulphidesignificantlysatisticallydecreasedinperiodofoestrusandrisedindioestrus.Thisobservationsuggestpossibledualcharacterofmechanismofinteractionbetweenmalesandfemalesduringmatingperiodincludingbothluringbutmaybealsorepellentsignalexistence.ContrarytoGoodwinetal.(1979)observation,noparabenswerefoundinexaminedsamples.Thefurtherstudyincludingthe

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behavioraltestsareindicatedtoverifysemiochemicalactivitiesofidentifiedsubstancesandgenuinenessofthesisaboutluring/repelingmechanisminvolvedinmatingbehaviorindogs.POSTER15:ScentCommunicationinBankVoles:Doesmaleinvestmentinfluencefemaleresponse?

H.Coombes1,Stockley1,R.J.Beynon2&J.L.Hurst1

1MammalianBehaviourandEvolutionGroup,UniversityofLiverpool,Liverpool,UK;2ProteinFunctionGroup,UniversityofLiverpool,Liverpool,UK

Olfactorycommunicationplaysakeyroleinmanymammaliansocialinteractionsandmanyanimalsinvestheavilyindepositingscentsignals.However,themolecularcompositionofscentmarks,whyanimalinvestinscentsignalsandbehaviouralresponseofconspecificstoindividualscentcuesispoorlyunderstoodinmostnon-modelspecies.Scentmarkingisimportantinvolesocialbehaviourandscentsignalsareakeysourceofinformationaboutconspecifics.Inthisstudy,weexaminescentmarkingbehaviourinbankvoles(Myodesglareolus).Weinvestigatewhethermalebankvolesinvestdifferentiallyincompetitivescentmarking,bothintermsofthenumberofmarksdepositedandtheproteincompositionofthesescentmarks.Wealsoexplorewhetherindividualmalescentmarkselicitdifferentbehaviouralresponsesfromfemalevoles,andwhetherfemalesshowapreferencebetweenmalesaccordingtotheirinvestmentinscentmarking.Understandingthecontextinwhichscentsignalsareproduced,theirmolecularcompositionandthebehaviouralresponsestheystimulate,eachprovideimportantinsightrequiredtounderstandhowchemicalcommunicationinfluencessocialbehaviour.POSTER16:ExploringVOCsfromharbourporpoiseexcreta

M.Gabirot1,B.Bouchard1,A.Célérier1,L.IJsseldijk2&S.Campagna1

1CEFE-CNRS(Montpellier,France);2UtrechUniversity(Netherlands)

Cetaceansaredescendantsofland-livingmammalswhichreturnedtowaterforafullyaquaticlife.Thissingularevolutionaryhistoryisaccompaniedbyaprofoundreorganizationofthechemosensorysystem(olfactionandgustation).Intoothedwhalesparticularly,thisisillustratedbythelossofanatomicalstructuresasolfactorybulbsandnerve.Howeverseveralreportsbasedonbehaviouralexperimentswithbottlenosedolphinsdemonstratethepresenceofaneffectivechemicalperceptioninfeedingaswellassocialcontext.Thusifchemicalsareinvolvedinsocialinteractionsincetaceans,informativecompoundsshouldbepresentintheexcretaoftheseanimals.Forthisreason,wehavelaunchedtheanalysisofthevolatileorganiccompounds(VOCs)releasedfromtheurineandfaecesofdeadharbourporpoise(Phocoenaphocoena).Wehavedevelopedforthatpurposeanovelandrobustmethodologyusingsolidphasemicroextraction(SPME).ThecomparisonofVOCsfoundamongtheexcretafromindividualsofdifferentages,sexesandhealthstatuswiththosepresentintheexcretafromterrestrialmammalswillallowustodetectthepresenceofsocialsemiochemicalsinmarinemammals.Thisinnovativeworkcouldalsohavemajor

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implicationsinmonitoringthehealthstatusofanimalsbasedonthepossiblepresenceofdiseasemarkersintheexcreta.

POSTER17:Putativepheromoneofsnowleopard,Pantherauncia(Schreber,1775)S.Das1,P.L.Das1,2&M.P.Sarkar1

1DepartmentofBotany(CentreofAdvancedStudy),ChemicalsignalandLipidomicslab.,UniversityofCalcutta,Kolkata700019,India2ZooBiologist,PNHZP,Darjeeling,WB,India

Chemicalcharacterizationofurineofsnowleopardwascarriedouttoinvestigateitspheromonalsignificance.Pantheraunciasyn.Unciauncia(Schreber,1775),acrepuscularandsolitaryendangeredfelid,isthenativeofNorthEasternHimalayanmountainrangeofIndiansubcontinent.Observationsonurinewereconductedonsemi-captiveanimalsofPadmajaNaiduHimalayanZoologicalPark,WestBengal,India(27°03'30.1"N88°15'14.4"E).Solidphasemicroextraction(SPME)ofheadspacevolatilesoftheurinefromsnowleopardwereexaminedbygaschromatography–massspectrometry.About32lowcarboncompoundswithvariousfunctionalgroupslikealcohols,aldehydes,ketones,phenol,amines,somecyclichydroxycompoundsandasulphurcontainingcompoundhavebeenidentified.Thevolatilomeofurineofsnowleopardrevealedthepresenceof6-methyl-5-hepten-2-one,theprincipalcomponentoflemongrassoilandthemajorpheromonalcomponentofredfox,Vulpesvulpes.Wedetectedthisspecificaromabysmelling,conductedonabouttwentyvolunteerswhensprayedoverfilterpaper.VariousbranchedandunbranchedsaturatedandunsaturatedfattyacidsfromC12-C18wereidentifiedfromthelipid,extractedfromurine.However,significantvariationincompositionofvolatilomeandlipidicfractionofurineofsnowleopardfromtiger,leopard,lionandAfricancheetahmightbeindicativefortheirdifferentstrategyandnatureofterritorialmarking.Itmightberelevanttotheirdifferentoriginindifferentecologicalenvironment.

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Listofdelegates:Surname First name Email address Affiliation Abbate Jessie jessie.abbate@gmail.com Laboratoire MIVEGEC, Institut de Recherche

pour le Développement, Montpellier, France Albone Eric eric@clifton-scientific.org Clifton Scientific Trust, UK

Allen Caroline caroline.allen@newcastle.ac.uk Institute of Neuroscience, Newcastle University, Newcastle Upon Tyne, Newcastle

Allen Tanesha tanesha.allen@zoo.ox.ac.uk WildCRU-Department of Zoology, University of Oxford

Apps Peter peterjapps@gmail.com Botswana Predator Conservation Trust

Baudouin Jean-Yves Jean-Yves.Baudouin@u-bourgogne.fr

Developmental Ethology and Cognitive Psychology, CSGA, Universiré de Bourgogne Franche-Comté, CNRS, Inra

Beynon Robert R.Beynon@liverpool.ac.uk University of Liverpool, UK

Bossuyt Franky fbossuyt@vub.be Amphibian Evolution Lab, Biology Department, Vrije Universiteit Brussel (VUB), Pleinlaan 2, B-1050 Brussels, Belgium

Cann Paul paul.cann@edu.mnhn.fr UMR 8576 CNRS/Université de Lille1/USC INRA 1409

Chabi Malika m.chabicenci@gmail.com UGSF- UMR 8576- USC INRA

Coombes Holly H.Coombes@liverpool.ac.uk Mammalian Behaviour and Evolution Group, University of Liverpool, Liverpool, UK

Das Subhadeep subhabiology@gmail.com Department of Botany, University of Calcutta

Davidson Amanda Mammalian Behaviour and Evolution Group, Institute of Integrative Biology, University of Liverpool, Leahurst Campus, Neston, CH64 7TE, United Kingdom

Dehnhard Martin dehnhard@izw-berlin.de Leibniz Institute for Zoo and Wildlife Research, Berlin, Germany

Devaraj Sankar Ganesh

Sankarganesh.eco@gmail.com Laboratory of Animal Behavior, Physiology and Welfare, Animal and Food Sciences Department, Texas Tech University, Lubbock, TX 79409

Durairaj Rajesh r.durairaj@group-irsea.com Research Institute in Semiochemistry and Applied Ethology

Dzięcioł Michał michaldzieciol@wp.pl Wroclaw University of Environmental and Life Sciences, Department of Reproduction and Clinic of Farm Animals, 50-366 Wrocław, Plac Grunwaldzki 49, Poland.

Ferdenzi Camille camille.ferdenzi@inserm.fr Lyon Neuroscience Research Center, CNRS UMR5292, INSERM U1028, University Claude Bernard Lyon 1, Lyon, France

Fialová Jitka jitka.fialova@natur.cuni.cz Faculty of Science, Charles University, Prague, Czech Republic

Fu Xiaoyan Xiaoyanfu@wustl.edu Department of Neuroscience, Washington University in St. Louis, USA

Gabirot Marianne marianne.gabirot@gmail.com CEFE-CNRS, Montpellier, France

Gervasi Stephanie sgervasi@monell.org Monell Chemical Senses Center

Ghosh Biswatosh biswabios.atgc@gmail.com Department of Zoology, Bidhannagar College, Govt. of West Bengal, Salt Lake City, Kolkata 700064, India.

Gómez-Baena Guadalupe v52gobag@uco.es

Centre for Proteome Research. University of Liverpool. United Kingdom.

Grau Paricio Carlos c.grau@group-irsea.com Research Institute in Semiochemistry and Applied Ethology Apt, France

Harris Rachel rachel.l.harris@duke.edu Department of Evolutionary Anthropology, Duke University, Durham NC, USA

Havlíček Jan jhavlicek@natur.cuni.cz National Institute of Mental Health, Klecany, Czech Republic

Hurst Jane J.Hurst@liverpool.ac.uk University of Liverpool, UK

Janssenswillen Sunita Sunita.Janssenswillen@vub.be Amphibian Evolution Lab, Biology Department, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, Belgium

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Joly Alexandre joly.alexandre14@gmail.com Unité de Glycobiologie Structurale et Fonctionnelle (UGSF), UMR8576, CNRS/Université de Lille 1/USC INRA1409 cité scientifique, Bâtiment C9 75655 Villeneuve d'ascq CEDEX

Kaneko Yayoi ykaneko7946@gmail.com Tokyo University of Agriculture and Technolgy

Kean Eleanor keanef@cardiff.ac.uk School of Biosciences, Cardiff University, Cardiff, UK

Kotenkova Elena ekotenkova@gmail.com A.N. Severtsov Institute of Ecology and Evolution RAS, Moscow, Russian Federation

Koyama Sachiko sakoyama@indiana.edu Indiana University, Medical Sciences, Bloomington, IN, U.S.A

Laktionova Tatiana tatita.laktionova@gmail.com A.N. Severtsov Institute of Ecology and Evolution, Moscow, Russian Federation

Le Danvic Chrystelle chrystelle.ledanvic@allice.fr ALLICE, Research and Development Department

Liu Dingzhen dzliu@bnu.edu.cn College of Life Sciences, Beijing Normal University

Loxley Grace Centre for Proteome Research, Institute of Integrative Biology, University of Liverpool, UK

Maex Margo Margo.Maex@vub.be Amphibian Evolution Lab, Biology Department, Vrije Universiteit Brussel (VUB), Pleinlaan 2, B-1050 Brussels, Belgium

Maita Minoru a2216009@iwate-u.ac.jp Department of Biological Chemistry and Food Sciences, Iwate University, Iwate, Japan

Mason Robert (Bob)

masonr@science.oregonstate.edu

Department of Integrative Biology, Oregon State University

McGlone John Texas Tech University

Mirza Reehan reehanm@nipissingu.ca Department of Biology and Chemistry, Nipissing University

Miyazaki Masao mmasao@iwate-u.ac.jp Iwate University, JAPAN

Munger Steven D. steven.munger@ufl.edu 1Center for Smell and Taste, 2Department of Pharmacology & Therapeutics and 3Department of Medicine, Division of Endocrinology, Diabetes and Nutrition, University of Florida, Gainesville, FL, 32610, USA.

Nagnan-LeMeillour

Patricia patricia.Le-Meillour@univ-lille1.fr

CNRS UMR 8576 - INRA 1409

Olsson Mats mats.j.olsson@ki.se Department of Clinical Neuroscience, Karolinska Institutet, Stockholm, Sweden

Petrulis Aras Neuroscience Institute, Georgia State University

Poirier Alice alice.poirier@pgr.anglia.ac.uk Anglia Ruskin University

Pounder Kieran Mammalian Behaviour and Evolution Group, Institute of Integrative Biology, University of Liverpool, Leahurst Campus, Neston, L64 7TE, United Kingdom

Prescott Mark mp1608@liverpool.ac.uk Centre for Proteome Research, Institute of Integrative Biology, University of Liverpool

Roberts S. Craig craig.roberts@stir.ac.uk Division of Psychology, University of Stirling, UK

Rowland Hannah Max Planck Institute for Chemical Ecology, Jena, Germany

Santoro Massimo msantoro@markes.com Markes International Ltd, Llantrisant, UK

Schaal Benoist benoist.schaal@u-bourgogne.fr CNRS centre des sciences du goût

Schulte Bruce bruce.schulte@wku.edu Western Kentucky University

Schulte Lisa M. Lisa.Maria.Schulte@vub.be Amphibian Evolution Lab, Biology Department, Vrije Universiteit Brussel (VUB), Pleinlaan 2, B-1050 Brussels, Belgium

Sugianto Nadine Adrianna

nadine.sugianto@zoo.ox.ac.uk WildCRU-Department of Zoology, University of Oxford

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Suzuki Chiharu g0417031@iwate-u.ac.jp Department of Biological Chemistry and Food Sciences, Faculty of Agriculture, Iwate University, Japan

Tognetti Arnaud arnaud.tognetti@gmail.com Institute for Advanced Study in Toulouse, 21 allée de Brienne, 31015 Toulouse – France

Treer Dag dtreer@vub.ac.be, dtreer@vub.be

Amphibian Evolution Lab, Biology Department, Vrije Universiteit Brussel (VUB), Pleinlaan 2, B-1050 Brussels, Belgium

Trester Annika G.M.Loxley@liverpool.ac.uk Institute of Integrative Biology, University of Liverpool, UK

Tsunoda Marie maririntay13@gmail.com Carnivore Ecology and Conservation Research Group, Tokyo University of Agriculture and Technology

Van Bocxlaer Ines Ines.Van.Bocxlaer@vub.be Amphibian Evolution Lab, Biology Department, Vrije Universiteit Brussel (VUB), Pleinlaan 2, B-1050 Brussels, Belgium

Van Moorleghem Charlotte Charlotte.VanMoorleghem@uantwerpen.be

Laboratory for Functional Morphology, Department of Biology, University of Antwerp, Universiteitsplein 1, B-2610 Wilrijk, Belgium

Voznessenskaya Vera veravoznessenskaya@gmail.com

A.N.Severtsov Institute of Ecology & Evolution, Moscow

Weiß Brigitte M. Brigitte.schloegl@uni-leipzig.de

Institute of Biology, University of Leipzig, Talstrasse 33, D-04103 Leipzig, Germany

Wyatt Tristram tristram.wyatt@zoo.ox.ac.uk

Department of Zoology, University of Oxford