Posted on Authorea 14 Aug 2020 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.159741548.88179477 | This a preprint and has not been peer reviewed. Data may be preliminary. efclttdb hi clgclrlaei h e niomn;eooia ees stecnetthat concept the is release ecological predators as environment; may such growth, that new population boom their the to founding demographic limitations in a top-down the from show release from released Excoffier species often ecological expand invasive in are their many species typically (reviewed introduced habitat, Invasions by pattern new facilitated temporal a 2015). be of and Lau, colonization spatial & successful predictable to After (Colautti response a investigate originates in to following opportunities it conditions tractable population(s) provide local as locations to adaptation new range. into rapidly local their expanded adapt recently expand have can they that taxa as species Invasive many encounter they that regimes show selection studies new These the rapidly more. often many (Rollins and and toads frequently 2016) cane Friesen, occurs as (Avalos & adaptation divergent Tigano honeybees local as 2011; necessary systems that Whitlock (Prentis be in know & environment to adaptation novel (Yeaman now thought a flow We time of gene the colonization high 2002). providing after with locations, (Lenormand, systems those evolve homogenizing in in the to even history from adaptation long free local a populationsthose local of isolated flowwith for degrees fairly gene in high high Traditionally, by only of present questions conditions. effects evolutionary be environmental and to differing ecological expected were between to adaptation interface adapt the populations bridged how long have exploring adaptation local of Studies these INTRODUCTION in adaptation local world-wide of North studies in ongoing expansion complements birds. of invasive and signatures and date genomic to of dispersive evolved survey comprehensive have highly This may most starlings the population. American is young North starlings evolutionarily genotype-environment in American and against In adaptation wide-ranging precipitation Local this and/or introduction. in divergence. temperature even Despite range-wide since with rapidly and size correlated genome- scale. genetic negligible are population continental moderate of polymorphisms effective background a single-nucleotide a in a only ~200 at show increase that even starlings found dramatic American we outliers a North associations FST suggest individuals, few 180 models and approximately rate and and of dispersal differentiation bottleneck, population high geographic founding species’ a low this from found with starting reduced-representation Consistent we using history, populations. differentiation this starling expansion and facilitated winter-season Genome-wide diversity rapid that 17 genomic continent. of factors of the (ddRADseq) selective patterns on sequencing and/or investigated species We genome demographic bird identify expansion. numerous can successful and and populations widespread rapid and most individuals the starling of across one comparisons pro- demographic becoming through America, ( both North differentiate starlings may throughout European environments novel selection. in local spread and and cesses colonize that species invasive of Populations Abstract 2020 14, August 2 1 America Hofmeister Natalie North in and starling invasive European the panmictic in largely variation genetic of correlates Environmental ainlWllf eerhCenter Research Wildlife National University Cornell tal., et 1 ct Werner Scott , 07,seledtot(Willoughby trout steelhead 2017), tru vulgaris Sturnus 2 n ryLovette Irby and , tal. et 1 eeitoue oNwYr n19 n usqetyspread subsequently and 1890 in York New to introduced were ) 08.W otnet n vdnefrrpdlocal rapid for evidence find to continue We 2008). , tal. et tal., et 1 05,sikeak (Lescak sticklebacks 2015), , 08,de ie(Pfeifer mice deer 2018), tal et tal. et tal., et ,2009). ., 2015), , 2018), Posted on Authorea 14 Aug 2020 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.159741548.88179477 | This a preprint and has not been peer reviewed. Data may be preliminary. rgntdnab.I te od,salnsta irt nesenNrhAeialkl xeinegreater to origin experience likely likely molt America less in North -90 be eastern differences of in may the migrate west south explains that collected starlings the better words, feathers longitude in shorter other starling that In migrate starlings nearby. found or that originated sampled populations: they disperse hypothesized individuals starling may same Instead, initially the U.S. among they from distances. western feathers Giltz, and the long used & survey, in migrate models (Burtt genetic starlings The flyways Previous this that 2020). within Hobson, for indicate 1982). distances & origin migratory (Dolbeer, Fischer, molt (Werner, in migrate distances variation of populations considerable Models allstarling is not 1977). there genetic of somebut that patterns indicate influenced America, also in studies North have might populations In dispersal some and that migration variation. ongoing possible expansion, is 1953). starling it the (Kessel, starling but During record Portland, earlier colonization, without the to after of persisted years introduced none U.S. that few been western America, indicate a The Records having the PA North than Allegheny, successful 1921). more in 2014). Gabrielson, (1875), second-most survived & starlings Graham, Canada Kalmbach the introductions 1915; & of Quebec, with (Forbush, 1889 (1872), (Bitton establishment (1897), in OH year MA successful OR Cincinnati, each Springfield, first in km and the introduced 91 (1897), were as to populations accepted up several widely expanding but is by Mexico introduction generations northern 1890 few from range a currently within starlings continent million (Linz 200 Alaska estimated southern an 2019). America, to Hallatschek, North & In (Gralka just ( spread be species. invasive may and starling chance establishment Regardless, European species’ invasive 2011). The an Leimu, changes & in adaptation, (Uller selection to invasiveness as pre-requisite explain important a not as as do viewed also alone often diversity can is genetic accelerates populations conditions in range introduced new native in the diversity The in genetic variants although genetic 2008). 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All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.159741548.88179477 | This a preprint and has not been peer reviewed. Data may be preliminary. Lnma azeg 02 sn h vr estv oa”sto lgmn r-es h bioinformatics The pre-sets. alignment of 2.2.8 set version local” bowtie2 using sensitive genome “very reference the the using to 2012) mapped Salzberg, were & reads Individual (Langmead stacks. in the option an ref-map passed to reads sequences met: assembled were We conditions adaptors. following indexing the Illumina intact if contain and an only not trimmed contained reads We filter, retained chastity 2500. we Illumina HiSeq the steps, (Catchen filtering used Illumina 1.19 subsequent then an v We In stacks on (http://hannonlab.cshl.edu/fastx_toolkit). in reads FASTX-Toolkit commands single-end the process_radtags 100bp, using quality sequenced We for Thrasher filtered P2. and in Peterson P1 associated described of restriction-site adaptors 95% protocol double-digested, as using of the in Following the markers quantified dataset following NY). frozen was genomic York, DNA kit reduced-representation sample New and a DNeasy each Scientific, generated Qiagen PA) of Fisher we a York, (Thermo (2012), concentration using fluorometer DNA Miltex, extracted 2.0 NY). was Qubit (Integra York, DNA a New punches and (Qiagen, ice, biopsy dry protocol on using manufacturer’s shipped sampled were Samples was & ethanol. 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No reuse without permission. — https://doi.org/10.22541/au.159741548.88179477 — This a preprint and has not been peer reviewed. 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Data may be preliminary. eioadClrd—n l te apiglctos(al ) id olce nteesouthwestern 2020); these Hobson, in & Fischer, collected (Werner, analysis Birds function discriminant 2). by (Table states differentiated those locations have to to sampling assigned appear also other F are U.S. all higher states western the Colorado—and the on in and based Starlings counterparts Mexico could eastern population. driver their migration that primary from F to in the subtly assigned was are expect differences movement and movements would starling regional collected that If we birds and suggests 2020). then dispersal survey Hobson, differentiation, genetic juvenile & of Fischer, this both (Werner, in variation that genetic sampled and influence starlings continent-wide, same spatial uniform the of not from possibility the data about isotope inferences Feather make to evidence 2011; genetic (Lee, and sorting. fitness isotopic in records, beetles shifts historical invasive combine lasting and 2008) to (Duckworth, lead bluebirds (Lombaert can Western in sorting expansion spatial some range which facilitate Williams of in can 2019; 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Data may be preliminary. pedo naieseis n ua omnaimmyspotetbihetadsra,a hw in shown as spread, and establishment support may commensalism success (Ravinet human invasion sparrows inhibit and house to as species, seem invasive just not of role be do may spread a world-wide, composition distributed suggests genetic (Szucs now invader work the (Edelaar beetles (Briski avian but population other flour another establishing success, However, parakeets, of the invasion monk of studies of size experimental driver 2018). the as a in important as as al. pressure such et propagule expansion, highlighted adaptation (Leydet range that corals rapid al., evidence in in supporting little et even find in as populations, invasions expanding selection expansion, expanding some for rapidly this in of adaptation facilitate studies rapid Some may of evidence systems. world-wide growing young starlings the extremely to in that contribute variation confirm results genetic populations Our observed introduced shape precipitation, and may and al., native precipitation temperature et of (Hofmeister in and resequencing extremes temperature On whole-genome by in in explained invasions. of differences variability be both their results can in despite preliminary isolation-by-distance variation selection yet and for genetic of drift, evidence starling signatures from continents, strong reveal selection both the disentangle associations Australia, genotype-environment to In attempts dynamics, invasion complicate sampled. effects areas founder similar and ge- (Cardilini despite explains of variation America, distance number environmental North geographic greater does much where than a Australia, better increased employing F in starlings survey has subdivisions in genotyping-by-sequencing haplotype differentiation population A mitochondrial netic three novel indicates edge. a (Rollins markers range invasion carrying SNP this Australian Australia at Western the (Rollins only during in site Australia adapted rapidly starlings Western rapidly introduction adult in have the of front may in expansion proportion front flow the living expansion of gene still the edge that at those the shown from on had starlings differentiated that (Rollins work have confirm low Earlier variation is sequence isolation-by-distance. populations chondrial of concur- explains starling patterns nearly distance Australian environmental significant continent among not and that but structuring colonized geographic invasion—which tion (Cardilini that there Australian invasion—found patterns the American our starlings. genetic North in American supports the starlings North evidence with in on This rently variation project genetic variation. 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(Table tend movements U.S. longer western dertake the in starlings ST cosalAsrla ouain sa re fmgiuehge hnteeuvln F equivalent the than higher magnitude of order an is populations Australian all across 07 n miia oki upe (Baltazar-Soares guppies in work empirical and 2017) tal., et 05.Peaatto ntentv ag rslcindrn rnpr a aiiaethe facilitate may transport during selection or range native the in Pre-adaptation 2015). nprep in tal., et ). priori a 08 n omnmns(Cohen mynas common and 2018) tal., et d o eoe n inl fpplto tutr.Ti sampling This structure. population of signals any recover not —do 00.Salnsi h utainrneso usata popula- substantial show range Australian the in Starlings 2020). 9 tal., et tal., et 09,adpyoegahcpten fmito- of patterns phylogeographic and 2009), tal., et 08.Freape eei oteek in bottlenecks genetic example, For 2018). tal., et 09.Ivso ilgsshv long have biologists Invasion 2019). 09,adrvee cosalien across reviewed and 2019), tal., et 01.I at starlings fact, In 2011). tal. et tal., et 00.Global 2020). , ST ne across index 06:the 2016): Posted on Authorea 14 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159741548.88179477 — This a preprint and has not been peer reviewed. Data may be preliminary. otrnso ilgclivso:itouto rmAi n ipra nErp ftetpot gudgeon topmouth the of Genomic Europe (2019) in C dispersal and Eizaguirre Asia Z´ahorsk´a, RE, from AS, introduction Gozlan Tarkan invasion: ( biological J, a Cote of S, footprints Blanchet M, Baltazar-Soares C honeybee. Li Africanized H, pumpkinseed Pan invasive A, and Avalos native of tendency dispersal and Boldness Tool. (2017) ( Search MG Fox Alignment AC, Local Rooke T, Basic Ashenden (1990) DJ Lipman EW, Biology Myers Molecular W, of Miller W, Gish SF, Altschul raw and 10.5061/dryad.j424f07), PRJNA545151). (doi: (Bioproject REFERENCES Dryad SRA on archived NCBI are in scripts archived above data the sequencing accompany that files data https://github.com/nathofme/radseq-NAm All GitHub: on archived are scripts All fel- following STATEMENT the ACCESSIBILITY by DATA Fellowship. supported Ornithology Ivy also Edsall the was Fellowship, and N.R.H. Graduate Fellowship, Treman-Williams Student N.R.H.) Fellowship, Graduate Mellon to Paul Graduate W. Walcott EEB (all University Charles Andrew Fund Cornell the Research lowships: the Grant, Student and Athena Award, Graduate Ornithology Research Feeny Society of P. Ornithological Lab American Cornell the the Grant, by Research supported was work This advice valuable provided Aguillon FUNDING Stepfanie New Walsh- and Jennifer Mexico, Berv, methods. here. New Jacob bioinformatic used Hampshire, Hooper, samples on California, Daniel New collected Arizona, Campagna, who Nevada, in Leonardo Wisconsin, Nebraska, personnel and Emond, Washington, Services Missouri, Texas, Wildlife Carolina, Kansas, Agriculture’s North Iowa, York, of Illinois, Department U.S. Idaho, the Colorado, to paper. grateful wrote are IJL We NRH, data, analyzed NRH samples, ACKNOWLEDGEMENTS contributed SJW research, designed IJL NRH, reveal that populations. studies young CONTRIBUTIONS and recent AUTHOR dispersive and (Schweizer other in vertebrates dispersal complement even American evolve of results North can studies in Our adaptation without loci variation flow Bay particular genes. genetic and gene 2015; carry variables how understand incipient climate that fully explore of between individuals cannot we evidence associations the we find Here but of we diversity, landscape, migration starlings. and the American differentiation novel across in North genetic changes establishment low in and of colonization associations of background adaptive genotype-environment consequences to a genetic lead On the that explore environments. processes to us ecological allow and populations demographic Invasive of range populations. wider invading much in a evolution consider (Liebl Africa should in may we invasions expression that supported have gene may also in al., shifts (Marin here changes starlings—epigenetic et species described to plastic invasive similar ones and/or quite of the shifts system expansion like sparrow—a epigenetic and invaders variation, establishment of genetic the studies to support Empirical addition in 2019). how, Blackburn, show & (Cardador species bird suoabr parva Pseudorasbora gibbosus Lepomis 03 u o eesrl nAsrla(Sheldon Australia in necessarily not but 2013) tal., et :i pta otn raigspro invaders? superior creating sorting spatial is ): 08.Fnly esgetta u td dst hs ugsigta ai local rapid that suggesting those to adds study our that suggest we Finally, 2018). , aueCommunications Nature 215 ). tal. et oeua Ecology Molecular 403–410. , 21)Asf eetv we uigrpdeouino etebhvori an in behaviour gentle of evolution rapid during sweep selective soft A (2017) , 29 1–9. , 71-85. , 10 tal., et 08.Tkntgte,rcn oksuggests work recent together, Taken 2018). qai Invasions Aquatic tal., et 09.I h elsuidhouse well-studied the In 2019). , 12 311-320. , Journal tal., et Posted on Authorea 14 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159741548.88179477 — This a preprint and has not been peer reviewed. Data may be preliminary. obe A(92 irto atrsfraeadsxcasso lcbrsadStarlings. and Blackbirds genetic of details: classes sex the and in age is for devil patterns Ornithology The Migration invasion. (2015) (1982) to RA HD contributions Dolbeer Gillette its FA, and Cang populations J, introduced Braasch in variation SR, Anderson KM, Dlugosch envi- using adaptation local uncover to method FST-based new A (2015) variables. OE ronmental Gaggiotti P, Villemereuil de natural G differentiation, Abecasis 2156–2158. for A, Auton evidence P, invasion: Danecek during evolution adaptation. Contemporary local (2015) and JA selection, Lau RI, Colautti signatures identify to analysis redundancy of Evaluation (2018) E Bazin adaptation. MGB, local Starling. 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North in ) , 53 28-46. , oeua Ecology Molecular , , 16 56 oeua clg Resources Ecology Molecular ehd nEooyadEvolution and Ecology in Methods 345-353. , 1189-1199. , 1–8. , Heredity rceig fteRylSceyB Society Royal the of Proceedings Bird-Banding oeua Ecology Molecular ora fZoology of Journal tal. et , 22 , 80 oeua Ecology Molecular 21)Tevratcl omtadVCFtools. and format call variant The (2011) 3124–3140. , Science tal. et 519–525. , , ciohrstristis Acridotheris 48 21)Gnmcsgaso eeto rdc climate-driven predict selection of signals Genomic (2018) 259-271. , , , , 359 185 , 24 11 295 1999–2017. , 83–86. , 587–11. , ilgclInvasions Biological , 254–260. , 6 , 17 1248–1258. , 431–449. , . , LSONE PLoS 281 oeua Ecology Molecular 20141385. , , 21 , h Condor The 7 1295-1309. , e38145. , rnir nEooyand Ecology in Frontiers Bioinformatics , 24 ora fField of Journal , 101 2095–2111. , 451–454. , Sturnus , 27 , Posted on Authorea 14 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159741548.88179477 — This a preprint and has not been peer reviewed. Data may be preliminary. eprr dpaint ua-lee aiaswti h aiernecnpooeinvasions. promote can range native the within Applications habitats human-altered to adaptation temporary V package Ravigne B, R Facon RA, Hufbauer data. Directions. raster Future with and KE Solutions, modeling Lotterhos Practical JL, and Kelley analysis S, Geographic Hoban raster: (2012) J 2.0-12. Etten version frequencies. van allele RJ, from Hijmans adaptation local Ed,). of identification Harmon, Robust (L (2013) America GC tests Coop Mantel T, Gunther the Dismantling ex- (2013) range F of Evolution Rousset genetics population G, the Guillot tip can heterogeneity Environmental contemporary (2019) on pansions. flow O gene Hallatschek and M, dispersal Gralka of effects multifarious The (2007) AP adaptation. Hendry SE, Forde D, Garant O’Meara, (B studies association ecological and landscape for adap- package R multilocus An Ed,). detecting LEA: (2015) for O Francois methods E, Frichot Comparing (2018) associations. DL genotype-environment Urban multivariate HH, with Co. Wagner tation Printing JR, Potter Lasky & BR, Wright Forester Boston, starling. The (1915) EH Forbush . (2017) RJ Hijmans SE, Fick A Tayeh from axyridis. inference RA, Harmonia demographic Hufbauer Robust B, (2013) Facon M Foll VC, data. Sousa SNP E, and Huerta-Sanchez genomic I, Dupanloup L, Excoffier Systematics and Evolution, Ecology, R of Hufbauer Review suggests Carrete V, parakeet TF, Ravigne monk Wright selection. A, the convergent Estoup JC, of of Senar range possibility MA, invasive the global Russello and the ML, origin 2164-2716. across Avery geographic diversity restricted AG, genetic common Silva Shared ecology.a Da (2015) and EA, JL evolution Tella Hobson in M, S, force Roques underappreciated P, an Edelaar flow: Evolution gene & Non-random Ecology (2012) in Trends DI Bolnick P, STRUC- method. visualizing Edelaar Evanno for the program implementing and website and a output HARVESTER: STRUCTURE TURE (2011) BM vonHoldt DA, Earl Expansion. Range a of Dynamics the and Naturalist Strategies Dispersal Adaptive (2008) RA Duckworth rco ,ShvleS,Buhr ,Faci 21)TsigfrAscain ewe oiadEnviron- and Loci between Models. Associations Mixed for Factor Testing Latent (2013) Using O Francois Gradients G, mental Bouchard SD, Schoville E, Frichot nentoa ora fClimatology of Journal International ehd nEooyadEvolution and Ecology in Methods , eLife , 195 , 4 172 ucinlEcology Functional , 336–344. , 5 205-220. , , 89–101. , S4–S17. , 8 e44359. , urn Biology Current LSGenetics, PLOS olCi :Nw1k pta eouinciaesrae o lblln areas land global for surfaces climate resolution spatial 1-km New 2: WorldClim , , 27 21 tal. et tal. et 659–665. , tal. et tal. et , 434–443. , 21 . 21)Atrpgnclyidcdaatto oivd AA) con- (AIAI): invade to adaptation induced Anthropogenically (2011) , 21)I hr eei aao fBooia Invasion? Biological of Paradox Genetic a There Is (2016) 9 424–427. , 21)FnigteGnmcBsso oa dpain Pitfalls, Adaptation: Local of Basis Genomic the Finding (2016) 6 21)Ibedn ersinI ugdi h naieInsect Invasive the in Purged Is Depression Inbreeding (2011) e1003905. , h mrcnNaturalist American The 925–929. , , 47 51–72. , 12 osrainGntc Resources Genetics Conservation oeua ilg n Evolution and Biology Molecular oeua Ecology Molecular , 188 379–397. , , 27 ehd nEooyand Ecology in Methods oeua Ecology Molecular 2215–2233. , , , 30 4 359–361. , 1687–1699. , h American The eeisSoc Genetics Evolutionary Annual , 24 , Posted on Authorea 14 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159741548.88179477 — This a preprint and has not been peer reviewed. Data may be preliminary. avrZ,Tbm F rnadN 21)Ppr nRpcaefrgntcaayi fppltoswith populations of analysis genetic for package reproduction. R sexual an and/or Poppr: clonal, (2014) partially NJ clonal, Grunwald JF, Tabima ZN, Kamvar aisyM rch ,Lwo ,Fls 21)RDane n nRDtutr:pplto inference population fineRADstructure: 1–6. and RADpainter data. 385-400. (2016) The RADseq D 34, invasion: Falush Ecology, from D, Biological Functional Lawson (2019) E, (epi)genome. Trucchi C M, the Vieira Malinsky CK, of Ghalambor side P, hidden Gibert the S, Maury of D, influence Barloy J, Genitoni P, Marin B Facon spectra. ladybird A, invasive frequency Estoup SNP E, using Lombaert changes size range population a Exploring throughout (2015) methylation 47 Y-X DNA Fu of X, Patterns Liu (2013) songbird. LB introduced Martin an CL, of Richards, expansion AW, Schrey AL, (2007) Liebl WJ Bleier LB, impacts Penry far-reaching SM, with Gaulker species inference HJ, structure Homan population GM, affect Linz strongly thresholds frequency datasets. allele Mediterranean genomic Minor Spanish (2017) with CJ the Battey along genetic EB, expansion Linck reveals range RAD-Seq with Host-targeted associated (2018) patagonica ME Oculina Hellberg coast. coral M, the Ribes R, in Coma changes CGB, Grupstra KP, Leydet selection. J islands. Catchen natural SL, to Bassham limits EA, the Lescak and 2. flow Bowtie Gene with (2002) alignment T gapped-read Lenormand Fast (2012) SL Salzberg B, Langmead Birds. Cavity-Nesting Native on Effect Their and , Starlings America. European North (2003) W in Koenig Starling European the of migration and 49–67. Distribution (1953) B Kessel States United the in Starling the of Value Economic markers. , (1921) genetic Gabrielson IN of and analysis ER, multivariate Kalmbach, the for package R a 24 adegenet: (2008) T Jombart In: America. North in Jarpas. Starlings structure. (2017) with population A dealing of Jernelov for analysis program in permutation and multimodality matching and cluster switching a CLUMPP: label (2007) NA Rosenberg M, Jakobsson AS Chaine microcosms. rule. ciliate D, bad-disperse” native Legrand in “good-stay, S, strategies the Jacob dispersal and Flexible (2012) quality KL environmental Evans 1024–1032. 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All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159741548.88179477 — This a preprint and has not been peer reviewed. Data may be preliminary. rthr K tpesM onlyP(00 neec fPplto tutr sn utlcsGenotype Multilocus Using Structure Population invasive of in Inference evolution (2000) P Adaptive Data. Donnelly (2008) M, AJ Stephens JK, Lowe Pritchard selection. DM, Richardson natural EE, of Dormontt analogue JRU, species. spatial Wilson the PJ, as Prentis sorting Spatial (2019) Australia. and TA Ecology Africa Perkins South dispersal-related BL, of of sorting vulgaris) Phillips spatial (Sturnus Context-dependent starlings (2018) invasive al the et C in Berthouly-Salazar traits J, Ed,). Roux Singh, Le (N DJ, Flow Phair Gene Strong of VC Face the Sousa in S, Adaptation Laurent effective SP, estimated Pfeifer with Ed,). structure Orlando, population spatial (L Visualizing Species surfaces. 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Team Core R Genetics , , rnsi ln Science Plant in Trends oeua Ecology Molecular 12 24 , 26 , 155-163. , 3223-3231. , 7 rnsi clg Evolution & Ecology in Trends 841–842. , aueCommunications Nature e37135–11. , , 155 aueRvesNeuroscience Reviews Nature , 28 945–959. , 659–669. , , 24 2264–2276. , tal. et , 13 tal. et 288–294. , 20)PNHRvrin7 mrvdpyoeei re,orthologs trees, phylogenetic improved 7: version PANTHER (2009) , tal. et 21)TeEouinr itr fNbak erMc:Local Mice: Deer Nebraska of History Evolutionary The (2018) 8 1–8. , rceig fteRylSceyB Society Royal the of Proceedings , 29 21)Slcino iohnra ainsOcr between Occurs Variants Mitochondrial on Selection (2016) , 233–242. , oeua ilg n Evolution and Biology Molecular 48 14 oeua Ecology Molecular 94–100. , uli cd Research Acids Nucleic oeua ilg n Evolution and Biology Molecular , 20 2307–2317. , , , bioRxiv, 285 38 , D204–D210. , 33 20181246. , 342451. 995–1007. , , 35 Theoretical 792–806. , Molecular rnsin Trends Bioin- Posted on Authorea 14 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159741548.88179477 — This a preprint and has not been peer reviewed. Data may be preliminary. etnJ,Hnate B omn A(04 eei slto yevrneto itne hc pattern which common? distance: or most environment is by flow isolation Genetic gene (2014) of AA Hoffmann SB, Hangartner JP, wolves. Sexton grey in variation functional driven R environmentally inbreeding Harrigan of J, role Robinson potential RM, the Schweizer success. revisited: invasion Invasions the of in Paradox of interactions Genetic environment success The Invasion X (2017) (2007) L Lachmuth bottleneck. GR K, genetic Allen Schrieber drastic OD, a Seeman despite PC, terrestris, Bombus Brunner bumblebee, range R, native Schmid-Hempel their in P, experiment species. 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R Leimu T, Uller 172185. , rceig fteNtoa cdm fSciences of Academy National the of Proceedings , oeua Ecology Molecular 65 lblCag Biology Change Global , 1897–1911. , 4 137–138. , tal. et , Evolution 18 21)Ahg-efrac optn ole o eaens n principal and relatedness for toolset computing high-performance A (2012) eoeResearch Genome Bioinformatics 1560–1573. , , 17 tal. et tal. et , 3478–3485. , 68 tal. et 1–15. , 21)Tree atr n eeunigo 00gnsreveal genes 1040 of resequencing and capture Targeted (2015) 21)Rpdaatv vlto nnvlevrnet acts environments novel in evolution adaptive Rapid (2017) ilRev Biol rceig fteNtoa cdm fSciences of Academy National the of Proceedings , 20)PNHR irr fPoenFmle and Families Protein of Library A PANTHER: (2003) 28 asrdomesticus Passer , 15 13 3326–3328. , e Phytologist New 2129–2141. , , , 92 97 oeua Ecology Molecular LSOne PLoS 939-952. , 5948–5953. , δ tru vulgaris Sturnus 2 H, Heredity δ 13 noAustralia. into ) oeua Ecology Molecular C, . , 185 oeua Ecology Molecular δ 15 , )taigo otoii for origin molt of tracing N) 99 , 1087–1099. , 25 ouain nWestern in populations ) 414–422. , 357–379. , .Sc pnSci Open Soc. R. , 25 , 17 2144–2164. , Molecular 675. , , 114 , , Posted on Authorea 14 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159741548.88179477 — This a preprint and has not been peer reviewed. Data may be preliminary. in-north-america correlates-of-genetic-variation-in-the-invasive-and-largely-panmictic-european-starling- Table2.docx file Hosted coefficient inbreeding the and diversity, nucleotide heterozygosity, expected and F observed alleles, private of 1. 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Population vdnefricpetlclaatto.Rdnac nlssidct ht11SP (small SNPs 191 that indicate analyses Redundancy adaptation. local incipient for Evidence vial at available vial at available ST PC2 1.03% A D PC2 (1.03%) Withinsamples Withinpopulations Betweenpopulations −0.2 −0.1 0.0 0.1 0.2 . −0.2 ● ● ● ● ● ● −0.1 ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● Total ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● 0.0 ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● PC1 (1.07%) ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● https://authorea.com/users/350824/articles/475552-environmental- https://authorea.com/users/350824/articles/475552-environmental- ● ST ● 0.1 ● PC1 1.07% ● ● 0.2 mn oain speetdaoetedaoa,addre rycolors gray darker and diagonal, the above presented is locations among ● ● 0.3 0.4 315 158 156 df ● 1 ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● 0.5 WI WA TX NY NV NM NH NE NC MO KS IL ID IA CO CA AZ ● B

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 K=2 Sum ofsquares 391273 184622 205297 1354 16 MO NM WA CO NC NH CA NY NV NE KS AZ TX WI ID IA IL

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 K=3 variance components C Within samples Within populations Between populations 0 50 100 150 200 0 50 100 150 200 0 50 0100 50150 100200 150 200 0 50 100 150 200 0 50 0100 50150 100200 150 200 0 50 100 150 200 0 50 100 150 200 73.75 0.264 1242 1168 −1.0 −1.0 −1.0 −50 −50 −50 1200 1200 1200 −0.5 −0.5 −0.5 Variations betweenpopulation Variations betweenpopulation Variations betweenpopulation Variations betweensamples Variations betweensamples Variations betweensamples Variations withinsamples Variations withinsamples Variations withinsamples 0 0 0 % varianceexplained vec.sim vec.sim vec.sim vec.sim vec.sim vec.sim 0.0 0.0 0.0 1250 1250 1250 94.04 5.936 0.021 0.5 0.5 0.5 50 50 50 1.0 1.0 1.0 1300 1300 1300 Posted on Authorea 14 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159741548.88179477 — This a preprint and has not been peer reviewed. Data may be preliminary.

RDA2RDA2 −2 −1 0 1 2 −3 ● ● ● ● ● AZ ● ● ● ● ● BIO1 −2 CA ● RDA1 ● −1 ● ● ● ● ● ● ● RDA1 17 ● ● ● ● ● ● ● NV ● NM ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● TX ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● BIO7 ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● 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● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● 0 ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● 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● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● NH ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● WA ● ● ● ● ● ● ● ● ● elev ● ● ● ● ● ● ● ● ● ● ● BIO12 ● ● ● ● ● ● ● ● ● 1 ● BIO16 ● ● ● ● ● ● ● ● CO ● ● ● ● ●