IRCFREPTILES&AMPHIBIANS•22(3):128–131•SEP2015

The Cuban Blue Anole, Anolis allisoni Barbour 1928 (Squamata: Dactyloidae),

a New Nonnative Lizard Introduced in FloridaKenneth L. Krysko1, Claudia MacKenzie-Krysko1, Laurence L. Connor2, Yasel U. Alfonso1, and Leroy P. Nuñez1

1FloridaMuseumofNaturalHistory,UniversityofFlorida,Gainesville,Florida32611,USA(KLK:kenneyk@ufl.edu,YUA:anoles1983cuba@gmail.com,LPN:therizinosaur@yahoo.com)

2FloridaFishandWildlifeConservationCommission(ret.),Eustis,Florida32726,USA(llconnor@comcast.net)

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IRCF REPTILES & AMPHIBIANS • VOL15, NO 4 • DEC 2008 189TABLE OF CONTENTS

T A B L E O F C O N T E N T S

F E A T U R E A R T I C L E S

Chasing Bullsnakes (Pituophis catenifer sayi) in Wisconsin: On the Road to Understanding the Ecology and Conservation of the Midwest’s Giant Serpent ...................... Joshua M. Kapfer 190

The Shared History of Treeboas (Corallus grenadensis) and Humans on Grenada: A Hypothetical Excursion ............................................................................................................................Robert W. Henderson 198

R E S E A R C H A R T I C L E S

The Texas Horned Lizard in Central and Western Texas ....................... Emily Henry, Jason Brewer, Krista Mougey, and Gad Perry 204 The Knight Anole (Anolis equestris) in Florida

.............................................Brian J. Camposano, Kenneth L. Krysko, Kevin M. Enge, Ellen M. Donlan, and Michael Granatosky 212

C O N S E R V A T I O N A L E R T

World’s Mammals in Crisis ............................................................................................................................................................. 220 More Than Mammals ...................................................................................................................................................................... 223 The “Dow Jones Index” of Biodiversity ........................................................................................................................................... 225

H U S B A N D R Y

Captive Care of the Central Netted Dragon ....................................................................................................... Shannon Plummer 226

P R O F I L E

Kraig Adler: A Lifetime Promoting Herpetology ................................................................................................ Michael L. Treglia 234

C O M M E N T A R Y

The Turtles Have Been Watching Me ........................................................................................................................ Eric Gangloff 238

B O O K R E V I E W

Threatened Amphibians of the World edited by S.N. Stuart, M. Hoffmann, J.S. Chanson, N.A. Cox, R. Berridge, P. Ramani, and B.E. Young .............................................................................................................. Robert Powell 243

CONSERVATION RESEARCH REPORTS: Summaries of Published Conservation Research Reports ................................. 245 NATURAL HISTORY RESEARCH REPORTS: Summaries of Published Reports on Natural History ................................. 247 NEWBRIEFS ...................................................................................................................................................................................... 248 EDITORIAL INFORMATION ..................................................................................................................................................... 251 FOCUS ON CONSERVATION: A Project You Can Support ............................................................................................... 252

Front Cover. Shannon Plummer.Totat et velleseque audant mo estibus inveliquo velique rerchil erspienimus, quos accullabo. Ilibus aut dolor apicto invere pe dolum fugiatis maionsequat eumque moditia erere nonsedis ma sectiatur ma derrovitae voluptam, as quos accullabo.

Back Cover. Michael KernTotat et velleseque audant mo

estibus inveliquo velique rerchil erspienimus, quos accullabo. Ilibus

aut dolor apicto invere pe dolum fugiatis maionsequat eumque

moditia erere nonsedis ma sectia-tur ma derrovitae voluptam, as

IRC

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REPTILES & AMPHIBIANSC O N S E R V AT I O N A N D N AT U R A L H I S T O R Y

Copyright©2015.KennethL.Krysko.Allrightsreserved.

WWW.IRCF.ORG/REPTILESANDAMPHIBIANSJOURNAL

Anoline lizards, Anolis(sensu lato;FamilyDactyloidae),areoneofthemostspeciesrichgroupsofterrestrialverte-

brateswith399currentlyrecognizedspecies(Nicholsonetal.2012;UetzandHošek2015).Becauseanolesarehighlyvariableinmorphology(ConantandCollins1998;Schettino1999;Camposano2011)andhavetheabilitytochangecolorinresponsetoexternalstimuli(Hadley1931;Weber1983),diagnosingspeciescanbedifficult,especiallywhendealingwithintroducedpopulations. ThestateofFlorida,USA,hasmoreintroducedamphib-iansandreptilesthananyotherplaceintheworld(Kryskoetal.2011a).ElevenspeciesofanoleshavebeendocumentedinFlorida,onlyoneofwhichisnative(GreenAnole,A. caroli-nensis[Voigt1832])andallbutone(JeremieAnole,A. coele-stinusCope1862)ofthenonnativetaxaareestablishedandreproducing(Kryskoetal.2011a,2011b).Inthispaper,wedocumenttheeleventhintroducedanole,theCubanBlue

Anole, Anolis allisoniBarbour1928,inFlorida.Anolis allisoni isnativetoBelize(IslasdelaBahia),Cuba,andHonduras(IslasdeBarbareta,Guanaja,Morat,Roatán,Utila,theCayosCochinosintheIslasdelaBahıa),andalsohasbeenintro-ducedintoLaCeiba,Atlantida(onthenorthernHondurancoast),andinQuintanaRoo,Mexico(McCranieandKöhler2015).Juvenilesandadultfemalesaregreenincolor,whereasadultmalesturnblueanteriorly(uniquewithinAnolis;butmales fromHondurashavenecks, limbs,and lowerbod-ies that remainbright green) andhave a reddishdewlap(Barbour1928;Lee2000).Anolis allisonireadilyhybridizeswiththeCubanGreenAnole,A. porcatusGray1840,inCuba(SchwartzandHenderson1991;Gloretal.2004),andA. porcatusisbelievedtohybridizewiththenativeA. carolinen-sisinitsintroducedrangeinsouthernFlorida(Kolbeetal.2007).Allthreeofthesecloselyrelatedspeciesareassignedtothe Anolis carolinensisseries(Losos2009).

Materials and MethodsSpecimen acquisition.—On 5 July 2013 at 0901 h, asingleadultmaleAnolis allisoniwasobservedonthebackofaresident’shouseonMagdaleneManorDrive,Tampa,Hillsborough County, Florida (Fig. 1; 28.07457°N,82.47916°W,DatumWGS84;15melev.).Adigitalimage(photographicvoucherUF-Herpetology170513)wasdepos-ited in theDivisionofHerpetology,FloridaMuseumofNaturalHistory,UniversityofFlorida.SpeciesidentitywasconfirmedbyJoseph.P.Burgess. After additional sightings by the homeowners,KLKandCMKvisitedthissiteon18August2013.Althoughnoadult male A. allisoniwasobserved,fivegreen-coloredjuve-nileanoleswerecollected(Fig.2;UF-Herpetology170869–170873) from the same side of the house.The residentinformedusthatheobservedthemaleA. allisoni the same day shortlyafterwedepartedthesite,andthatneighborsbelieveasimilarunusuallycoloredlizardwasobservedontheirhomes

I N T R O D U C E D S P E C I E S

Fig. 1.AdultmalenonnativeCubanBlueAnole(Anolis allisoni)pho-tographedinTampa,HillsboroughCounty,Florida,on5July2013.PhotographbyTonyHenneke.

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aswell.On21September2013,KLKandCMKvisitedthissiteagain,butafterintensivesearchesdidnotobserveamaleA. allisoni. Laboratory techniques.—WeobtainedDNA isola-tions from the five Anolis specimensusingZRGenomicDNA™TissueMicroprepKit (ZymoResearch, LLC).UsingtotalcellularDNAasatemplateandpolymerasechainreaction(PCR)methodology(Saikietal.1988),DNAwasamplifiedandsequencedfollowingGloretal. (2004)for

nicotinamideadeninedinucleotidedehydrogenasesubunit2(ND2)usingprimersL4437b(Maceyetal.1997)andH5730(Gloretal.2004).PCRwasconductedin25ml reac-tions:9.5ml H2O,12.5mlGoTaq

® MasterMix(PromegaCorp,Madison,Wisconsin,USA),1.0mleachprimer(10mM),and1.0ml DNA template. PCR parameters included initialdenaturingat94°Cfor3min,followedby35cyclesofamplification:denaturingat94°Cfor1min,annealingat52°Cfor1min,andextensionat72°Cfor1min,fol-

IRCFREPTILES&AMPHIBIANS•22(3):128–131•SEP2015KRYSKO ET AL.

Fig. 2.GreenAnoles(Anolis carolinensis)collectedon18August2013fromthesamesiteasthenonnativeCubanBlueAnole(Anolis allisoni)inTampa,HillsboroughCounty,Florida.NotethatA–D=UF-Herpetology170869–170873,respectively.PhotographsbyKennethL.Krysko.

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lowedbyafinalextensionat72°Cfor7min.Threeml of eachPCRproductwereelectrophoresedona1%agarosegel,visualizedwithGelRedTMstaining(BiotiumInc.,Hayward,California,USA), and comparedwith aDNA standard.Sequencefilesreceivedforourspecimensfromtheautomatedsequencer(GenomicsDivision,InterdisciplinaryCenterforBiotechnologyResearch,UniversityofFlorida)wereeditedasnecessarywithGeneious software (ver.6.1,createdbyBiomatters<http://www.geneious.com>).GenBankaccessionnumbersforourspecimensareKP174772–KP174776. Phylogenetic analyses.—We downloaded DNAsequencedataforND2fromGenBankfor45samplesfromthe Anolis carolinensisseries,includingA. allisoni, A. altitu-dinalis, A. carolinensis, A. isolepis A. porcatus, and A. smarag-dinus(Gloretal.2004).Alongwithourfivespecimens,allsequenceswereassembledusingtheMusclealgorithmandmanuallyeditedasnecessarywithGeneioussoftware. Atotalof1,038basepairs(bp)ofsequencedatawereanalyzed.RelationshipsamonghaplotypeswereestimatedusingMaximumLikelihood(ML)methodologywiththeTamura-Nei model, complete deletion mechanism, nucleo-

tide substitution, nearest-neighbor interchange heuristicmethod,verystrongbranchswapfilter,and1,000nonpara-metricbootstrapreplicates(Felsenstein1985)toassessnodesupportusingMEGAversion6(Tamuraetal.2013).Themostcrediblesupportofphylogeneticrelationshipswascon-finedtonodeswherenonparametricbootstrapvalues≥70%(HillisandBull1993;Felsenstein2004).

ResultsOurMLanalysis(Fig.3)producedaphylogenysimilartothatofGloretal.(2005).OfthefiveAnolis specimenswecollected,fourhaplotypeswererecoveredandallwerenestedwithin theA. carolinensis clade. These samples are most closely related to the nearest sample locality in Citrus County, Florida, and all A. carolinensis are most closely related to A. porcatusfromwesternCuba.

DiscussionInthepast,introducedspecieshavebeenreportedinthelit-eraturebyvariousmeans;somebyimproperlywritingthespeciesnamewithoutprovidinganyevidenceormeansof

Fig. 3.MaximumLikelihoodphylogenyforGreenAnoles(Anolis carolinensis)(UF-Herpetology170869–170873highlightedinred)collectedinTampa,HillsboroughCounty,Florida,on18August2013.Notethatvalues(≥70%)abovemajornodesrepresentbootstrapsupport.

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verification,orbyproperlyvoucheringaspecimen(preferred)and/oradistinguishablephotographinasystematiccollec-tionwithassociateddata(seeKryskoetal.2011a).Inthispaper,wefollowtheproperwayofdocumentinganewintro-ducedspeciesbyprovidingaclearphotographicvoucher,butalsogoastepfurtherbyattemptingtodetermineitsstageofintroduction(i.e.,ifitisestablished)usingmolecularanalysis.Althoughourvoucherphotographclearlyillustratesanadultmale Anolis allisoni,ourmoleculardatasuggestthatthefiveothergreenanolescollectedatthesamesitehavemtDNAof native A. carolinensis(Fig.3),andthereforenocurrentevidencesupportsanestablishedpopulationofA. allisoni. However,wecannotdetermineifA. allisonihybridizedinthewildwithnativeA. carolinensisbecauseourdataconsistofmtDNA,whichismaternallyinherited.Nonetheless,A. allisoniwaseitherreleasedorescapedfromanearbyenclosureandcurrentlyrepresentsastage-2introduction(afterColauttiandMacIsaac2004).

AcknowledgmentsWethankTonyandBarbaraHennekeforreportingthenon-nativeCubanBlueAnoleandallowingustocollectspecimenson their property; Gustav Paulay and David Reed provided laboratoryspace.

Literature CitedBarbourT.1928.ReptilesfromtheBayIslands.Proceedings of the New England

Zoölogical Club10:55–61(“Anolis allisonisp.nov.,”pp.58–59).

Camposano,B.J.2011.Morphologicalspeciesverificationandgeographicdistribu-tion of Anolis(Sauria:Polychrotidae)inFlorida.M.S.Thesis,UniversityofFlorida, Gainesville.

Colautti,R.I.andH.J.MacIsaac.2004.Aneutralterminologytodefine‘invasive’species. Diversity and Distributions10:135–141.

Conant,R.andJ.T.Collins.1998.A Field Guide to Reptiles and Amphibians of Eastern and Central North America.Thirdedition,expanded.HoughtonMifflin, Boston, Massachusetts.

Felsenstein,J.1985.Confidencelimitsonphylogenies:Anapproachusingthebootstrap.Evolution39:783–791.

Felsenstein, J. 2004. Inferring Phylogenies. Sinauer Associates, Sunderland, Massachusetts.

Glor,R.E.,M.E.Giford,A.Larson,J.B.Losos,L.RodríguezSchettino,A.R.ChamizoLara,andT.R.Jackman.2004.Partialislandsubmergenceandspe-

ciationinanadaptiveradiation:AmultilocusanalysisoftheCubangreenanoles. Proceedings of the Royal Society B271:2257–2265.

Glor,R.E.,J.B.Losos,andA.Larson.2005.OutofCuba:OverwaterdispersalandspeciationamonglizardsintheAnolis carolinensissubgroup.Molecular Ecology 14:2419–2432.

Hadley,C.E.1931.Colorchangesinexcisedandintactreptilianskin.Journal of Experimental Zoology58:321–331.

Hillis,D.M.andJ.J.Bull.1993.Anempiricaltestofbootstrappingasamethodforassessingconfidenceinphylogeneticanalysis.Systematic Biology42:182–192.

Kolbe,J.J.,R.E.Glor,L.RodríguezSchettino,A.ChamizoLara,A.Larson,andJ.B.Losos.2007.Multiplesources,admixture,andgeneticvariationinintro-duced Anolis lizard populations. Conservation Biology21:1612–1625.

Krysko,K.L.,J.P.Burgess,M.R.Rochford,C.R.Gillette,D.Cueva,K.M.Enge,L.A.Somma,J.L.Stabile,D.C.Smith,J.A.Wasilewski,G.N.KieckheferIII,M.C.Granatosky,andS.V.Nielsen.2011a.Verifiednon-indigenousamphibiansandreptiles inFlorida from1863through2010:Outliningtheinvasionprocessandidentifyinginvasionpathwaysandstages.Zootaxa 3028:1–64.

Krysko,K.L.,K.M.Enge,andP.E.Moler.2011b.Atlas of Amphibians and Reptiles in Florida.FinalReport,ProjectAgreement08013.FloridaFishandWildlifeConservation Commission, Tallahassee.

Lee, J.C.2000.A Field Guide to the Amphibians and Reptiles of the Maya World: The Lowlands of Mexico, Northern Guatemala, and Belize.ComstockPublishingAssociates,NewYork.

Losos,J.B.2009.Lizards in an Evolutionary Tree: Ecology and Adaptive Radiation of Anoles. University of California Press, Berkeley.

Macey,J.R.,A.Larson,N.B.Ananjeva,andT.J.Papenfuss.1997.Evolutionaryshiftsinthreemajorstructuralfeaturesofthemitochondrialgenomeamongiguanianlizards.Journal of Molecular Evolution44:660–674.

McCranie,J.R.andG.Köhler2015.Theanoles(Reptilia:Squamata:Dactyloidea:Anolis:Norops)ofHonduras.Systematics,distributionandconservation.Bulletin of the Museum of Comparative ZoologySPS(1):1–292.

Nicholson,K.E.,B.I.Crother,C.Guyer,andJ.M.Savage.2012.Itistimeforanewclassificationofanoles(Squamata:Dactyloidae).Zootaxa3477:1–108.

Peterson,J.A.andE.E.Williams.1981.Acasehistoryinretrogradeevolution:Theoncalineageinanolinelizards.II.Subdigitalfinestructure.Bulletin of the Museum of Comparative Zoology149:215–268.

RodríguezSchettino,L.(ed.).1999.The Iguanid Lizards of Cuba. University of Florida Press, Gainesville.

Saiki,R.K.,D.H.Gelfand,S.Stoffel,S.J.Scharf,R.Higuchi,G.T.Horn,K.B.Mullis,andH.A.Erlich.1988.Primer-directedenzymaticamplificationofDNAwiththermostableDNApolymerases.Science239:487–491.

Schwartz,A.andR.W.Henderson.1991.Amphibians and Reptiles of the West Indies: Descriptions, Distributions, and Natural History. University Press of Florida, Gainesville.

Tamura,K.,G.Stecher,D.Peterson,A.Filipski,andS.Kumar.2013.MEGA6:MolecularEvolutionaryGeneticsAnalysisVersion6.0.Molecular Biology and Evolution30:2725–2729.

Uetz,P.andHošek,J.2015.The Reptile Database.<www.reptile-database.org>.

Weber,W.1983.Photosensitivityofchromatophores.American Zoologist23:495–506.

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