The role of adaptive divergence in the formation of new species has been the subject of much recent debate. The most direct evidence comes from traits that can be shown to have diverged under natural selection and that now contribute to reproductive isolation. Here, we investigate differential adaptation of two fire-bellied toads (Anura, Bombinatoridae) to two types of aquatic habitat. Bombina bombina and B. variegata are two anciently diverged taxa that now reproduce in predator-rich ponds and ephemeral aquatic sites, respectively. Nevertheless, they hybridise extensively wherever their distribution ranges adjoin. We show in laboratory experiments that, as expected, B. variegata tadpoles are at relatively greater risk of predation from dragonfly larvae, even when they display a predator-induced phenotype. These tadpoles spent relatively more time swimming and so prompted more attacks from the visually hunting predators. We argue in the discussion that genomic regions linked to high activity in B. variegata should be barred from introgression into the B. bombina gene pool and thus contribute to gene flow barriers that keep the two taxa from merging into one.

Faculty of Veterinary Hygiene and Ecology University of Veterinary and Pharmaceutical Sciences Brno Czech Republic

Research Facility Studenec Institute of Vertebrate Biology Czech Academy of Sciences Brno Czech Republic

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Hereford J. A quantitative survey of local adaptation and fitness trade-offs. Am Nat. 2009;173: 579–588. 10.1086/597611 PubMed DOI

Schluter D. Ecology and the origin of species. Trends Ecol Evol. 2001;16: 372–380. 10.1016/s0169-5347(01)02198-x PubMed DOI

Schluter D. Evidence for ecological speciation and its alternative. Science. 2009;323: 737–741. 10.1126/science.1160006 PubMed DOI

Hendry AP. Ecological speciation! Or the lack thereof? Can J Fish Aquat Sci. 2009;66: 1383–1398.

Nosil P. Ecological Speciation Oxford: Oxford University Press; 2012.

Bierne N, Gagnaire P-A, David P. The geography of introgression in a patchy environment and the thorn in the thigh of ecological speciation. Curr Zool. 2013;59: 72–86.

Faria R, Renaut S, Galindo J, Pinho C, Melo-Ferreira J, Melo M, et al. Advances in ecological speciation: an integrative approach. Mol Ecol. 2014;23: 513–521. 10.1111/mec.12616 PubMed DOI

Filchak KE, Roethele JB, Feder JL. Natural selection and sympatric speciation in the apple maggot fly, Rhagoletis pomonella. Nature. 2000;407: 739–742. 10.1038/35037578 PubMed DOI

Linn C, Feder JL, Nojima S, Dambroski HR, Berlocher SH, Roelofs W. Fruit odor discrimination and sympatric host race formation in Rhagoletis. Proc Natl Acad Sci USA. 2003;100: 11490–11493. 10.1073/pnas.1635049100 PubMed DOI PMC

McGee MD, Schluter D, Wainwright PC. Functional basis of ecological divergence in sympatric stickleback. BMC Evol Biol. 2013;13: 277 10.1186/1471-2148-13-277 PubMed DOI PMC

Le Pennec G, Butlin RK, Jonsson PR, Larsson AI, Lindborg J, Bergström E, et al. Adaptation to dislodgement risk on wave-swept rocky shores in the snail Littorina saxatilis. PLOS ONE. 2017;12: e0186901 10.1371/journal.pone.0186901 PubMed DOI PMC

Johannesson B. Shell morphology of Littorina saxatilis Olivi: The relative importance of physical factors and predation. J Exp Mar Biol Ecol. 1986;102: 183–195. 10.1016/0022-0981(86)90175-9 DOI

Schluter D. Adaptive Radiation in Sticklebacks: Trade-Offs in Feeding Performance and Growth. Ecology. 1995;76: 82–90. 10.2307/1940633 DOI

Janson K. Selection and migration in two distinct phenotypes of Littorina saxatilis in Sweden. Oecologia. 1983;59: 58–61. 10.1007/BF00388072 PubMed DOI

Linn CE, Dambroski HR, Feder JL, Berlocher SH, Nojima S, Roelofs WL. Postzygotic isolating factor in sympatric speciation in Rhagoletis flies: Reduced response of hybrids to parental host-fruit odors. Proc Natl Acad Sci U S A. 2004;101: 17753–17758. 10.1073/pnas.0408255101 PubMed DOI PMC

Hatfield T, Schluter D. Ecological speciation in sticklebacks: environment-dependent hybrid fitness. Evolution. 1999;53: 866–873. 10.1111/j.1558-5646.1999.tb05380.x PubMed DOI

Feder JL, Opp SB, Wlazlo B, Reynolds K, Go W, Spisak S. Host fidelity is an effective premating barrier between sympatric races of the apple maggot fly. Proc Natl Acad Sci USA. 1994;91: 7990–7994. 10.1073/pnas.91.17.7990 PubMed DOI PMC

Rundle HD, Nagel L, Boughman JW, Schluter D. Natural selection and parallel speciation in sympatic sticklebacks. Science. 2000;287: 306–308. 10.1126/science.287.5451.306 PubMed DOI

Bay RA, Arnegard ME, Conte GL, Best J, Bedford NL, McCann SR, et al. Genetic coupling of female mate choice with polygenic ecological divergence facilitates Stickleback speciation. Curr Biol. 2017;27: 3344–3349.e4. 10.1016/j.cub.2017.09.037 PubMed DOI PMC

Feder JL, Roethele JB, Filchak KE, Niedbalski J, Romero-Severson J. Evidence for inversion polymorphism related to sympatric host race formation in the apple maggot fly, Rhagoletis pomonella. Genetics. 2003;163: 939–953. PubMed PMC

Meyers PJ, Doellman MM, Ragland GJ, Hood GR, Egan SP, Powell THQ, et al. Can the genomics of ecological speciation be predicted across the divergence continuum from host races to species? A case study in Rhagoletis. Philos Trans R Soc B Biol Sci. 2020;375: 20190534 10.1098/rstb.2019.0534 PubMed DOI PMC

Szymura JM. Analysis of hybrid zones with Bombina In: Harrison RG, editor. Hybrid zones and the evolutionary process. New York: Oxford University Press; 1993. pp. 261–289.

Nürnberger B, Baird SJE, Čížková D, Bryjová A, Mudd AB, Blaxter ML, et al. A dense linkage map for a large repetitive genome: discovery of the sex-determining region in hybridising fire-bellied toads (Bombina bombina and B. variegata). BioRxiv [Preprint]. 2020. bioRxiv 328633 [posted 9 Oct 2020; cited 21 Oct 2020]: [51 p.]. Available from: https://www.biorxiv.org/content/10.1101/2020.10.06.328633v1 PubMed DOI PMC

Yanchukov A, Hofman S, Szymura JM, Mezhzherin S, Morozov-Leonov SY, Barton NH, et al. Hybridization of Bombina bombina and B. variegata (Anura, Discoglossidae) at a sharp ecotone in Western Ukraine: comparisons across transects and over time. Evolution. 2006;60: 583–600. PubMed

Barton NH, Gale KS. Genetic analysis of hybrid zones In: Harrison RG, editor. Hybrid zones and the evolutionary process. Oxford: Oxford University Press; 1993. pp. 13–45.

Szymura JM, Barton NH. The genetic structure of the hybrid zone between the fire-bellied toads Bombina bombina and B. variegata: comparison between transects and between loci. Evolution. 1991;45: 237–261. 10.1111/j.1558-5646.1991.tb04400.x PubMed DOI

Kruuk LEB, Gilchrist JS, Barton NH. Hybrid dysfunction in Fire-Bellied Toads (Bombina). Evolution. 1999;53: 1611–1616. 10.1111/j.1558-5646.1999.tb05425.x PubMed DOI

Philippi D, Gollmann G. On the status of the fire-bellied toad, Bombina bombina, in Lobau (Vienna, Donau-Auen National Park). Acta ZooBot Austria. 2014;150/151: 25–32.

Gollmann B, Gollmann G. Die Gelbbauchunke: von der Suhle zur Radspur Bielefeld: Laurenti Verlag; 2002.

Nürnberger B, Barton NH, MacCallum C, Gilchrist J, Appleby M. Natural selection on quantitative traits in the Bombina hybrid zone. Evolution. 1995;49: 1224–1238. 10.1111/j.1558-5646.1995.tb04449.x PubMed DOI

Czopkowa G, Czopek J. The vascularization of the respiratory surfaces in Bombina bombina. Bull Acad Pol Sci Cl II. 1955;3: 313–318.

Hartel T. Movement activity in a Bombina variegata population from a deciduous forested landscape. North-West J Zool. 2008;4: 79–90.

Kovar R, Brabec M, Vita R, Bocek R. Spring migration distances of some Central European amphibian species. Amphib-Reptil. 2009;30: 367–378.

Boulenger GA. On two European species of Bombinator. Proc Zool Soc Lond. 1886;1886.

Lörcher K. Vergleichende bioakustische Untersuchungen an der Rot- und Gelbbauchunke, Bombina bombina (L.) und Bombina v. variegata (L.). Oecologia. 1969;3: 84–124. 10.1007/BF00344635 PubMed DOI

Rafińska A. Reproductive biology of the fire-bellied toads, Bombina bombina and B. variegata (Anura: Discoglossidae): egg size, clutch size and larval period length differences. Biol J Linn Soc. 1991;43: 197–210.

Kruuk LEB, Gilchrist JS. Mechanisms maintaining species differentiation: predator mediated selection in a Bombina hybrid zone. Proc R Soc Lond B. 1997;264: 105–110.

Reichwaldt E. Der Einfluss von Prädatoren auf die Kaulquappen hybridisierender Unken (Bombina, Discoglossidae, Amphibia): Entwicklung, Morphologie und Verhalten. Diploma Thesis, University of Munich. 1999.

Babbitt KJ, Baber MJ, Tarr TL. Patterns of larval amphibian distribution along a wetland hydroperiod gradient. Can J Zool. 2003;81: 1539–1552.

Richter-Boix A, Llorente GA, Montori A. A comparative study of predator-induced phenotype in tadpoles across a pond permanency gradient. Hydrobiologia. 2007;583: 43–56. 10.1007/s10750-006-0475-7 DOI

Werner EE, Skelly DK, Relyea RA, Yurewicz KL. Amphibian species richness across environmental gradients. Oikos. 2007;116: 1697–1712.

Woodward BD. Predator-prey interactions and breeding pond use of temporary pond species in a desert anuran community. Ecology. 1983;64: 1549–1555.

Smith DC, Van Buskirk J. Phenotyic design, plasticity, and ecological performance in two tadpole species. Am Nat. 1995;145: 211–233.

Relyea RA. Morphological and behavioural plasticity of larval anurans in response to different predators. Ecology. 2001;82: 523–540.

Lawler SP. Behavioural responses to predators and predation risk in four species of larval anurans. Anim Behav. 1989;38: 1039–1047.

Morin PJ. Predation, competition, and the composition of larval anuran guilds. Ecol Monogr. 1983;53: 119–138.

Azevedo-Ramos C, Van Sluys M, Hero J-M, Magnusson WE. Influence of tadpole movement on predation by odonate naiads. J Herpetol. 1992;26: 335–338.

Chovanec A. The influence of tadpole swimming behaviour on predation by dragonfly nymphs. Amphib-Reptil. 1992;13: 341–349.

Relyea RA. The Relationship between predation risk and antipredator responses in larval Anurans. Ecology. 2001;82: 541–554.

Dayton GH, Saenz D, Baum KA, Langerhans RB, DeWitt TJ, Lindström J. Body shape, burst speed and escape behavior of larval Anurans. Oikos. 2005;111: 582–591.

Werner EE, Anholt BR. Ecological consequences of the trade-off between growth and mortality-rates mediated by foraging activity. Am Nat. 1993;142: 242–272. 10.1086/285537 PubMed DOI

Cressler CE, King AA, Werner EE. Interactions between behavioral and life‐history trade‐offs in the evolution of integrated predator‐defense plasticity. Am Nat. 2010;176: 276–288. 10.1086/655425 PubMed DOI

Richardson JML. The relative roles of adaptation and phylogeny in determination of larval traits in diversifying Anuran lineages. Am Nat. 2001;157: 282–299. 10.1086/319196 PubMed DOI

Skelly DK, Werner EE. Behavioral and life-history responses of larval American toads to an odonate predator. Ecology. 1990;71: 2313–2322.

McCollum SA, Van Buskirk J. Costs and benefits of a predator-induced polymorphism in the gray treefrog Hyla chrysoscelis. Evolution. 1996;50: 583–593. 10.1111/j.1558-5646.1996.tb03870.x PubMed DOI

Laurila A, Kujasalo J, Ranta E. Predator induced changes in life history in two anuran tadpoles: effects of predators on diet. Oikos. 1998;83: 307–317.

Relyea RA, Werner EE. Quantifying the relation between predator-induced behavior and growth performance in larval anurans. Ecology. 1999;80: 2117–2124.

Steiner UK. Linking antipredator behaviour, ingestion, gut evacuation and costs of predator-induced responses in tadpoles. Anim Behav. 2007;74: 1473–1479. 10.1016/j.anbehav.2007.02.016 DOI

Orizaola G, Richter‐Boix A, Laurila A. Transgenerational effects and impact of compensatory responses to changes in breeding phenology on antipredator defenses. Ecology. 2016;97: 2470–2478. 10.1002/ecy.1464 PubMed DOI

Lima SC, Dill LM. Behavioral decisions made under the risk of predation: a review and prospectus. Can J Zool. 1991;268: 619–640.

Anholt BR, Werner E, Skelly DK. Effect of food and predators on the activity of four larval ranid frogs. Ecology. 2000;81: 3509–3521. 10.1890/0012-9658(2000)081[3509:EOFAPO]2.0.CO;2 DOI

Van Buskirk J. A comparative test of the adaptive plasticity hypothesis: Relationships between habitat and phenotype in Anuran larvae. Am Nat. 2002;160: 87–102. 10.1086/340599 PubMed DOI

Dijk B, Laurila A, Orizaola G, Johansson F. Is one defence enough? Disentangling the relative importance of morphological and behavioural predator-induced defences. Behav Ecol Sociobiol. 2016;70: 237–246. 10.1007/s00265-015-2040-8 DOI

Teplitsky C, Plénet S, Joly P. Tadpoles’ responses to risk of fish introduction. Oecologia. 2003;134: 270–277. 10.1007/s00442-002-1106-2 PubMed DOI

Kurali A, Pásztor K, Hettyey A, Tóth Z. Resource-dependent temporal changes in antipredator behavior of common toad (Bufo bufo) tadpoles. Behav Ecol Sociobiol. 2018;72: 91 10.1007/s00265-018-2503-9 DOI

McCollum SA, Leimberger JD. Predator-induced morphological changes in an amphibian: predation by dragonflies affects tadpole shape and color. Oecologia. 1996;109: 615–621. PubMed

Touchon JC, Warkentin KM. Fish and dragonfly nymph predators induce opposite shifts in color and morphology of tadpoles. Oikos. 2008;117: 634–640. 10.1111/j.0030-1299.2008.16354.x DOI

Van Buskirk J. The costs of an inducible defense in anuran larvae. Ecology. 2000;81: 2813–2821.

Relyea RA. Getting out alive: how predators affect the decision to metamorphose. Oecologia. 2007;152: 389–400. 10.1007/s00442-007-0675-5 PubMed DOI

Van Buskirk J. Natural variation in morphology of larval Amphibians: Phenotypic plasticity in nature? Ecol Monogr. 2009;79: 681–705.

Michel MJ. Spatial dependence of phenotype-environment associations for tadpoles in natural ponds. Evol Ecol. 2011;25: 915–932. 10.1007/s10682-010-9441-y DOI

Arendt J. Morphological correlates of sprint swimming speed in five species of spadefoot toad tadpoles: Comparison of morphometric methods. J Morphol. 2010;271: 1044–1052. 10.1002/jmor.10851 PubMed DOI

Van Buskirk J, Anderwald P, Lüpold S, Reinhardt L, Schuler H. The lure effect, tadpole tail shape, and the target of dragonfly strikes. J Herpetol. 2003;37: 420–424.

Johnson JB, Burt DB, DeWitt TJ. Form, function, and fitness: Pathways to survival. Evolution. 2008;62: 1243–1251. 10.1111/j.1558-5646.2008.00343.x PubMed DOI

Van Buskirk J, McCollum SA. Functional mechanisms of an inducible defence in tadpoles: morphology and behaviour influence mortality risk from predation. J Evol Biol. 2001;13: 336–347. 10.1046/j.1420-9101.2000.00173.x DOI

Teplitsky C, Plenet S, Léna J-P, Mermet N, Malet E, Joly P. Escape behaviour and ultimate causes of specific induced defences in an anuran tadpole. J Evol Biol. 2005;18: 180–190. 10.1111/j.1420-9101.2004.00790.x PubMed DOI

Kraft PG, Wilson RS, Franklin CE, Blows MW. Substantial changes in the genetic basis of tadpole morphology of Rana lessonae in the presence of predators. J Evol Biol. 2006;19: 1813–1818. 10.1111/j.1420-9101.2006.01185.x PubMed DOI

Innes-Gold AA, Zuczek NY, Touchon JC. Right phenotype, wrong place: predator-induced plasticity is costly in a mismatched environment. Proc R Soc B Biol Sci. 2019;286: 20192347 10.1098/rspb.2019.2347 PubMed DOI PMC

Van Buskirk J, McCollum SA, Werner EE. Natural selection for environmentally induced phenotypes in tadpoles. Evolution. 1997;51: 1981–1990. 10.1111/j.1558-5646.1997.tb05119.x PubMed DOI

Van Buskirk J, Relyea RA. Selection for phenotypic plasticity in Rana sylvatica tadpoles. Biol J Linn Soc. 1998;65: 301–328. 10.1111/j.1095-8312.1998.tb01144.x DOI

Van Buskirk J. Spatially heterogeneous selection in nature favors phenotypic plasticity in anuran larvae. Evolution. 2017;71: 1670–1685. 10.1111/evo.13236 PubMed DOI

Vorndran IC, Reichwaldt E, Nürnberger B. Does differential susceptibility to predation in tadpoles stabilize the Bombina hybrid zone? Ecology. 2002;83: 1648–1659.

Fijarczyk A, Nadachowska K, Hofman S, Litvinchuk SN, Babik W, Stuglik M, et al. Nuclear and mitochondrial phylogeography of the European fire-bellied toads Bombina bombina and Bombina variegata supports their independent histories. Mol Ecol. 2011;20: 3381–3398. 10.1111/j.1365-294X.2011.05175.x PubMed DOI

Baláž V, Vojar J, Civiš P, Šandera M, Rozínek R. Chytridiomycosis risk among Central European amphibians based on surveillance data. Dis Aquat Organ. 2014;112: 1–8. 10.3354/dao02799 PubMed DOI

Blooi M, Pasmans F, Longcore JE, Spitzen-van der Sluijs A, Vercammen F, Martel A. Duplex real-time PCR for rapid simultaneous detection of Batrachochytrium dendrobatidis and Batrachochytrium salamandrivorans in Amphibian samples. J Clin Microbiol. 2013;51: 4173–4177. 10.1128/JCM.02313-13 PubMed DOI PMC

Werner EE, McPeek MA. Direct and indirect effects of predators on two Anuran species along an environmental gradient. Ecology. 1994;75: 1368–1382. 10.2307/1937461 DOI

Bradley R, Terry M. Rank analysis of incomplete block designs. I. The method of paired comparisons. Biometrika. 1952;39: 324–345.

Bates D, Mächler M, Bolker B, Walker S. Fitting linear mixed-effects models using lme4. J Stat Softw. 2015;67: 1–48. 10.18637/jss.v067.i01 DOI

Winter B. Linear models and linear mixed effect models in R with linguistic applications. 2013. Available: https://arxiv.org/abs/1308.5499

McCoy MW, Bolker BM, Osenberg CW, Miner BG, Vonesh JR. Size correction: comparing morphological traits among populations and environments. Oecologia. 2006;148: 547–554. 10.1007/s00442-006-0403-6 PubMed DOI

R Core Team. R: A language and environment for statistical computing Vienna, Austria: R Foundation for Statistical Computing; 2018. Available: https://www.R-project.org/

Gosner KL. A simplified table for staging anuran embryos and larvae with notes on identification. Herpetologica. 1960;16: 183–190.

Fischer N. Performance of pure, natural hybrid and F1 Bombina tadpoles in semi-natural enclosures: Morphology, behaviour, growth, survival and development time. Diploma Thesis, University of Munich. 2006.

Eklöv P, Werner EE. Multiple predator effects on size-dependent behavior and mortality of two species of Anuran larvae. Oikos. 2000;88: 250–258.

Melvin SD, Houlahan JE. Tadpole mortality varies across experimental venues: do laboratory populations predict responses in nature? Oecologia. 2012;169: 861–868. 10.1007/s00442-012-2260-9 PubMed DOI

Van Buskirk J. Habitat partitioning in European and North American pond-breeding frogs and toads. Divers Distrib. 2003;9: 399–410. 10.1046/j.1472-4642.2003.00038.x DOI

Barton NH. Multilocus clines. Evolution. 1983;37: 454–471. 10.1111/j.1558-5646.1983.tb05563.x PubMed DOI

Barton NH, Bengtsson B-O. The barrier to genetic exchange between hybridising populations. Heredity. 1986;56: 357–376. 10.1038/hdy.1986.135 PubMed DOI

Sachdeva H, Barton NH. Introgression of a block of genome under infinitesimal selection. Genetics. 2018;209: 1279–1303. 10.1534/genetics.118.301018 PubMed DOI PMC

Arnegard ME, McGee MD, Matthews B, Marchinko KB, Conte GL, Kabir S, et al. Genetics of ecological divergence during speciation. Nature. 2014;511: 307–311. 10.1038/nature13301 PubMed DOI PMC

Rice WR, Hostert EE. Laboratory experiments on speciation: what have we learned in 40 years? Evolution. 1993;47: 1637–1653. 10.1111/j.1558-5646.1993.tb01257.x PubMed DOI

Nosil P, Harmon LJ, Seehausen O. Ecological explanations for (incomplete) speciation. Trends Ecol Evol. 2009;24: 145–156. 10.1016/j.tree.2008.10.011 PubMed DOI

Anholt BR, Werner EE. Interaction between food availability and predation mortality mediated by adaptive behavior. Ecology. 1995;76: 2230–2234.

Touchon JC, Robertson JM. You cannot have it all: Heritability and constraints of predator-induced developmental plasticity in a Neotropical treefrog. Evolution. 2018;72: 2758–2772. 10.1111/evo.13632 PubMed DOI

Barandun J, Reyer H-U. Reproductive Ecology of Bombina variegata: Development of Eggs and Larvae. J Herpetol. 1997;31: 107–110. 10.2307/1565337 DOI

Köhler SC. Mechanisms of partial reproductive isolation in a Bombina hybrid zone in Romania. Ph.D. Thesis, University of Munich. 2003.

Hartel T, Nemes S, Mara G. Breeding phenology and spatio-temporal dynamics of pond use by the Yellow-Bellied Toad (Bombina variegata) population: The importance of pond availability and duration. Acta Zool Litu. 2007;17: 56–63. 10.1080/13921657.2007.10512816 DOI

Newman RA. Effects of density and predation on Scaphiopus couchi tadpoles in desert ponds. Oecologia. 1987;71: 301–307. 10.1007/BF00377299 PubMed DOI

Relyea RA. Competitor-induced plasticity in tadpoles: Consequences, cues, and connections to predator-induced plasticity. Ecol Monogr. 2002;72: 523–540. 10.1890/0012-9615(2002)072[0523:CIPITC]2.0.CO;2 DOI

Lemcke C. Verwandtenerkennung bei Kaulquappen der Gelbbauchunke, Bombina variegata. Diploma Thesis, University of Munich. 2002.

Böll S. Ephemere Laichgewässer: Anpassungsstrategien und physiologische Zwänge der Gelbbauchunke (Bombina variegata) in einem Lebensraum mit unvorhersehbarem Austrocknungrisiko. Ph.D. Thesis, Julius-Maximilians-Universität, Würzburg. 2002.

Barandun J, Reyer H-U, Anholt B. Reproductive ecology of Bombina variegata: aspects of life history. Amphib-Reptil. 1997;18: 347–355. 10.1163/156853897X00404 DOI

Hantzschmann AM, Gollmann B, Gollmann G, Sinsch U. The fast–slow continuum of longevity among yellow-bellied toad populations (Bombina variegata): intrinsic and extrinsic drivers of variation. PeerJ. 2019;7: e8233 10.7717/peerj.8233 PubMed DOI PMC

Thiesmeier B. Fotoatlas der Amphibienlarven Deutschlands, Österreichs und der Schweiz Bielefeld: Laurenti Verlag; 2019.

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