Abstract
The bocagei clade is a species complex of dendrobatid frogs evidenced from recent molecular systematic studies. This lineage is endemic to the eastern Andean slopes and adjacent Amazonian lowlands in southern Colombia, Ecuador and northern Peru. Herein, we analyze the taxonomy of the bocagei clade by means of qualitative and quantitative morphological characters and call parameters. We recognize and provide accounts for six species: Hyloxalus bocagei, H. faciopunctulatus, H. maculosus, H. sauli and two new species. We also provide phylogeographic data. We formally assign the name of Hyloxalus bocagei only to populations from the surroundings of Volcán Reventador (Provincia Napo, Ecuador). Additionally, we describe the tadpoles of Hyloxalus bocagei, H. maculosus, and two new species, plus vocalizations of H. bocagei, H. maculosus, H. sauli, and two new species. Morphological variation was estimated from 16 morphometric characters using multivariate analysis. Adults of the two new species were indistinguishable using quantitative and qualitatively morphological characters. Nevertheless, molecular data, call parameters (dominant frequency, note repetition rate), and tadpole morphology supported the recognition of these two species as cryptic taxa. A current phylogenetic tree using molecular characters of five species of the bocagei clade is also provided including the maximum likelihood (ML) phylogeny and a chronogram of the group. Our phylogeny confirms the close relationship of members of the bocagei clade with Andean relatives of Hyloxalus and recurrent dispersal events from the Andes to the Amazon Basin in the late Miocene (< 10 MYA). Our data also corroborate that the sister lineage of bocagei clade includes the Andean H. subpunctatus clade. We anticipate that further detailed morphological and molecular data will be necessary to reveal the existence of additional cryptic species within the bocagei clade.
References
Acosta-Buenaño, N.A. (2002) Vocalización de 9 especies del género Colostethus (Anura: Dendrobatidae) del Ecuador. Tesis de Licenciatura, Quito, Pontificia Universidad Católica del Ecuador, 155 pp.
Akaike, H. (1973) A new look at the statistical model identification. IEEE Transactions on Automatic Control, 19, 716–723.
Altig, R. & McDiarmid, R.W. (1999) Body Plan: development and morphology. In: McDiarmid, R.W. & Altig, R. (Eds.) Tadpoles. The University of Chicago Press, Chicago, U.S.A., 295–335 pp.
Angulo, A. (2008) Hyloxalus maculosus. IUCN 2010. IUCN Red List of Threatened Species. [Web application] Version 2010.2. Available at: http://www.iucnredlist.org/apps/redlist/details/135881/0 (Accessed on 29 July 2010).
AmphibiaWeb. (2010) AmphibiaWeb: information on amphibian biology and conservation. [Web application] 2010. University of California Berkeley. Berkeley, California: AmphibiaWeb. Available at: http://amphibiaweb.org (Accessed 15 September 2010).
Andersson, L.G. (1945) Batrachians from east Ecuador collected 1937, 1938 by Wm Clarke-MacIntyre and Rolf Blomberg. Arkiv för Zoologi, 37, 1–88.
Bass, M.S., Finner, M., Jenkins, C.N., Kreft, H., Cisneros-Heredia, D.F., McCracken, S.F., Pitman, N.C.A., English, P.H., Swing, K., Villa, G., Di Fiore, A., Voigt, C.C. & Kunz, T.H. (2010) Global conservation significance of Ecuador’s Yasuní National Park, PLoS ONE, 5, 1-22.
Bolívar, W. & Rueda, J.V. (2004) Hyloxalus faciopunctulatus. IUCN 2010. IUCN Red List of Threatened Species. [Web application] Version 2010.2. Available at: http://www.iucnredlist.org/apps/redlist/details/55079/0 (Accessed on 13 July 2010).
Boulenger, G.A. (1882) Catalogue of the Batrachia Salientia s. Ecaudata in the collection of the British Museum. British Museum, London, England.
Bustamante, M.R., Ron, S.R. & Coloma, L.A. (2005) Changes in diversity of seven anuran communities in the Ecuadorian Andes. Biotropica, 37, 180–189.
Caldwell, J.P. & Lima, A.P. (2003) A new amazonian species of Colostethus (Anura: Dendrobatidae) with a nidicolous tadpole. Herpetologica, 59, 219–234.
Caldwell, J.P., Lima, A.P. & Biavati, G.M. (2002) Descriptions of tadpoles of Colostethus marchesianus and Colostethus caeruleodactylus (Anura: Dendrobatidae) from their type localities. Copeia, 166–172.
Castillo-Trenn, P. (2004) Description of the tadpole of Colostethus kingsburyi (Anura: Dendrobatidae) from Ecuador. Journal of Herpetology, 38, 600–606.
Castro, F., Rueda, J.V., Bolívar, W., Estupiñán, R.A. & Almendáriz, A. (2004) Hyloxalus sauli. IUCN 2010. IUCN Red List of Threatened Species. [Web application] Version 2010.2. Available at: http://www.iucnredlist.org/apps/redlist/details/55148/0 (Accessed on 29 July 2010).
Charif, R.A., Clark, C.W. & Fristrup, K.M. (2004) Raven 1.2 User’s Manual. Cornell Laboratory of Ornithology, Ithaca, New York, 191 pp.
Coloma, L.A. (1995) Ecuadorian frogs of the genus Colostethus (Anura: Dendrobatidae). Miscellaneous Publications of the Museum of Natural History University of Kansas, 87, 1–72.
Duellman, W.E. (1978) The biology of an Equatorial herpetofauna in Amazonian Ecuador. Miscellaneous Publications of the Museum of Natural History University of Kansas, 65, 1–352.
Duellman, W.E. (1988) Patterns of species diversity in anuran amphibian in the American tropics. Annals of the Missouri Botanical Garden, 75, 79-104.
Duellman, W.E. (2004) Frogs of the genus Colostethus (Anura: Dendrobatidae) in the Andes of northern Peru. Scientific Papers Natural History Museum the University of Kansas, 35, 1–49.
Duellman, W.E. & Pyles, R. (1983) Acoustic resource partitioning in anuran communities. Copeia, 1983, 639-649.
Drummond, A.J. & Rambaut, A. (2007) BEAST: Bayesian evolutionary analysis by sampling trees. BMC Evolutionary Biology, 7, 214.
Edwards, S.R. (1971) Taxonomic notes on South American Colostethus with descriptions of two new species (Amphibia, Dendrobatidae). Proceedings of the Biological Society of Washington, 84, 147–162.
Edwards, S.R. (1974a) A phenetic analysis of the genus Colostethus (Anura: Dendrobatidae). Ph.D. dissertation. Lawrence, University of Kansas.
Edwards, S.R. (1974b) Taxonomic notes on South American dendrobatid frogs of the genus Colostethus. University of Kansas, Natural History Museum, Occasional Papers, 30, 1–14.
Elmer, K.R., Dávila, J.A. & Lougheed, S.C. (2007) Cryptic diversity and deep divergence in an upper Amazonian frog, Eleutherodactylus ockendeni. BMC Evolutionary Biology, 7, 247.
Elmer, K.R. & Cannatella, D.C. (2008) Three new species of leaflitter frogs from the upper Amazon forests: cryptic diversity within Pristimantis “ockendeni” (Anura: Strabomantidae) in Ecuador. Zootaxa, 1784, 11-38.
Fouquet, A., Vences, M., Salducci, A.M., Marty, C., Blanc, M. & Gilles, A. (2007) Revealing cryptic diversity using molecular phylogenetics and phylogeography in frogs of the Scinax ruber and Rhinella margaritifera species groups. Molecular Phylogenetics and Evolution, 43, 567–582.
Frost, D.M. (2009) Amphibian Species of the World: an Online Reference. [Web application] Version 5.3 (12 February 2009). American Museum of Natural History. New York, USA. Available at: http://research.amnh.org/herpetology/amphibia/index.html. (Accessed 9 November 2009).
Funk, W.C., Angulo, A., Caldwell, J.P., Ryan, M.J. & Cannatella, D.C. (2008) Comparison of morphology and calls of two cryptic species of Physalaemus (Anura: Leiuperidae). Herpetologica, 64, 290–304.
Goebel, A.M., Donnelly, J.M. & Atz, M.E. (1999) PCR primers and amplification methods for 12S ribosomal DNA, the control region, cytochrome oxidase I, and cytochrome b in bufonids and other frogs, and an overview of PCR primers which have amplified DNA in amphibians successfully. Molecular Phylogenetics and Evolution, 11, 163–199.
Gosner, K.L. (1960) A simplified table for staging anuran embryos and larvae with notes on identification. Herpetologica, 16, 183–190.
Grant, T. & Ardila-Robayo, M.C. (2002) A new species of Colostethus (Anura: Dendrobatidae) from the eastern slopes of the Cordillera Oriental of Colombia. Herpetologica, 58, 252-260.
Grant, T., Frost, D.R., Caldwell, J.P., Gagliardo, R., Haddad, C.F.B., Kok, P.J.R., Means, D.B., Noonan, B.P., Schargel, W.E. & Wheeler, W.C. (2006) Phylogenetic systematics of dart-poison frogs and their relatives (Amphibia: Athesphatanura: Dendrobatidae). Bulletin of the American Museum of Natural History, 299, 1–262.
GraphPad Software. (2002-2005) Quick calcs, Online calculators for scientists. [Web application] Available at: http://www.graphpad.com/quickcalcs/ttest1.cfm. (Accessed January 2010)
Gray, P. & Cannatella, D.C. (1985) A new species of Atelopus (Anura, Bufonidae) from the Andes of northern Peru. Copeia, 910–917.
Guayasamin, J.M. & Trueb, L. (2007) A new species of glass frog (Anura: Centrolenidae) from the lowlands of northwestern Ecuador, with comments on centrolenid osteology. Zootaxa, 1447, 27–45.
Guindon, S. & Gascuel, O. (2003) A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Systematic Biology, 52, 696–704.
Hall, M.L., Samaniego, P., Le Pennec, J.L. & Johnson, J.B. (2008) Ecuadorian Andes volcanism: A review of Late Pliocene to present activity. Journal of Volcanology and Geothermal Research, 176, 1–6.
Heyer, W.R., García-López, J.M. & Cardoso, A.J. (1996) Advertisement call variation in the Leptodactylus mystaceus species complex (Amphibia: Leptodactylidae) with description of new sibling species. Amphibia-Reptilia, 17, 7–31.
Heyer, W.R. & Maxson, L.R. (1982) Neotropical frog biogeography: paradigms and problems. American Zoologist, 22, 397–410.
Hoorn, C. (1994) Fluvial palaeoenvironments in the intracratonic Amazonas Basin (Early Miocene-early Middle Miocene, Colombia). Palaeogeography, Palaeoclimatology, Palaeoecology, 109, 1–54.
Huelsenbeck, J.P. & Ronquist, F.P. (2001) MRBAYES: Bayesian inference of phylogenetic trees. Bioinformatics, 17, 754–755.
IUCN, Conservation International & NatureServe. (2006) Global Amphibian Assessment. [Web application] Available at: http://www.globalamphibians.org (Accessed 19 May 2009).
IUCN Standards and Petitions Working Group. (2008) Guidelines for using the IUCN Red List Categories and Criteria. [Web application] Version 7.0. (August 2008). Standards and Petitions Working Groups of the IUCN SSC Biodiversity Assesment Sub-Committe. Available at: http://intranet.iucn.org/webfiles/doc/SSC/RedList/RedListGuidelines.pd. (Accessed 15 May 2009).
Jiménez de la Espada, M. (1871) Faunae neotropicalis species quaedam nondum cognitae. Jornal de Sciencias Mathematicas Physicas e Naturaes, III, 54–65.
Jiménez de la Espada, M., Martínez, F., Almagro, M. & Isern, J. (1998) El gran viaje. Ediciones Abya-Yala, Quito, Ecuador, 250 pp.
Kohn, E. (2002) Natural Engagements and Ecological Aesthetics among the Ávila Runa of Amazonian Ecuador. Ph.D. dissertation, Department of Anthropology, University of Wisconsin-Madison, 495 pp.
Leviton, A.E. (1980) Museum acronyms¾second edition. Herpetological Review, 11, 93-102.
Maddison, W.P. & Maddison, D.R. (2009) Mesquite: a modular system for evolutionary analysis. [Web application] Available at: http://mesquiteproject.org. (Accessed March 2010).
Mendelson, J.R., 3rd, Lips, K.R., Gagliardo, R.W., Rabb, G.B., Collins, J.P., Diffendorfer, J.E., Daszak, P., Ibáñez, D.R., Zippel, K.C., Lawson, D.P., Wright, K.M., Stuart, S.N., Gascon, C., da Silva, H.R., Burrowes, P.A., Joglar, R.L., La Marca, E., Lotters, S., du Preez, L.H., Weldon, C., Hyatt, A., Rodríguez-Mahecha, J.V., Hunt, S., Robertson, H., Lock, B., Raxworthy, C.J., Frost, D.R., Lacy, R.C., Alford, R.A., Campbell, J.A., Parra-Olea, G., Bolaños, F., Domingo, J.J., Halliday, T., Murphy, J.B., Wake, M.H., Coloma, L.A., Kuzmin, S.L., Price, M.S., Howell, K.M., Lau, M., Pethiyagoda, R., Boone, M., Lannoo, M.J., Blaustein, A.R., Dobson, A., Griffiths, R.A., Crump, M.L., Wake, D.B. & Brodie, E.D., Jr. (2006) Biodiversity. Confronting amphibian declines and extinctions. Science, 313, 48.
Myers, C.W. & Duellman, W.E. (1982) A new species of Hyla from Cerro Colorado, and other tree frog records and geographical notes from western Panama. American Museum Novitates, 2752, 1–32.
Ojeda, D., Barbosa, C., Pinto, J., Cardona, M.C., Cuéllar, M., Cruz, S., de la Torre, L.S., Castañeda, J., Barrera, C.R., González, Y. & Alarcón, J.C. (2001) Ecosistemas. In: Leyva, P. (Ed.) El Medio Ambiente en Colombia. Instituto de Hidrología, Meteorología y Estudios Ambientales, IDEAM, Bogotá, Colombia, 279–346 pp.
Padial, J.M., Köhler, J., Muñoz, A. & De la Riva, I. (2008) Assessing the taxonomic status of tropical frogs through bioacoustics: geographical variation in the advertisement calls in the Eleutherodactylus discoidalis species group (Anura). Zoological Journal of the Linnean Society, 152, 353-365.
Palacios, W., Cerón, C., Valencia, R. & Sierra, R. (1999) Las formaciones naturales de la Amazonia del Ecuador. In: Sierra, R. (Ed.) Propuesta preliminar de un sistema de clasificación de vegetación para el Ecuador continental. Proyecto INEFAN/GERF-BIRF y Ecociencia, Quito, Ecuador, 109–118 pp.
Parmalee, J.R. (1999) Trophic ecology of a tropical anuran assemblage. Scientific Papers Natural History Museum the University of Kansas, 11, 53-74.
Peters, J.A. (1973) The frog genus Atelopus in Ecuador (Anura: Bufonidae). Smithsonian Contributions to Zoology, 145, 1–49.
Posada, D. (2008) jModelTest: Phylogenetic Model Averaging. Molecular Biology and Evolution, 25, 1253–1256.
Rambaut, A. & Drummond, A.J. (2007) Tracer v1.4. [Web application] Available from http://beast.bio.ed.ac.uk/Tracer.
Rivero, J.A. (1991) New Colostethus (Amphibia: Dendrobatidae) from South America. Breviora, 493, 1–28.
Rivero, J.A. & Almendáriz, A. (1991) La identificación de los Colostethus (Amphibia: Dendrobatidae) de Ecuador. Politécnica, XVI, 99–152.
Ron, S.R. (2001-2009) Anfibios de Parque Nacional Yasuní, Amazonía ecuatoriana. [Web application] Version 1.6 (2009). Museo de Zoología, Pontificia Universidad Católica del Ecuador. Quito, Ecuador. Available at: http://tropicalfrogs.net/yasuni/. (Accessed November 2009)
Ron, S.R., Guayasamin, J.M., Coloma, L.A. & Menéndez-Guerrero, P.A. (2008) Lista Roja de los Anfibios de Ecuador. [Web application] Version 1.0 (02 de mayo de 2008). Museo de Zoología, Pontificia Universidad Católica del Ecuador. Quito, Ecuador. Available at: http://www.puce.edu.ec/zoologia/sron/roja/. (Accessed November 2009)
Ron, S.R., Santos, J.C. & Cannatella, D.C. (2006) Phylogeny of the túngara frog genus Engystomops (= Physalaemus pustulosus species group; Anura; Leptodactylidae). Molecular Phylogenetics and Evolution, 39, 392–403.
Santos, J.C. (2002) Relaciones filogenéticas de Colostethus bocagei y Epipedobates tricolor (Anura: Dendrobatidae) inferidas de secuencias de genes mitocondriales 12S, 16S y Citocromo b. Tesis de Licenciatura, Quito, Pontificia Universidad Católica del Ecuador. 152 pp.
Santos, J.C., Coloma, L.A. & Cannatella, D.C. (2003) Multiple, recurring origins of aposematism in diet specialization in poison frogs. Proceedings of the National Academy of Sciences of the United States of America, 100, 12792–12797.
Santos, J.C., Coloma, L.A., Summers, K., Caldwell, J.P., Ree, R. & Cannatella, D.C. (2009) Amazonian amphibian diversity is primarily derived from Late Miocene Andean lineages. PLOS Biology, 7, 1–14.
Savage, J.M. (1978) Introduction. In: Jiménez de la Espada, M. Vertebrados del Viaje al Pacífico, Batracios. Society for the Study of Amphibians and Reptiles, pp. vii–xvi.
Savage, J.M. & Heyer, W.R. (1967) Variation and distribution of the tree-frog genus Phyllomedusa in Costa Rica, Central America. Beitrage zur Neotropischen Fauna, 5, 111–131.
Simões, P.I., Lima, A.P. & Farias, I.P. (2010) The description of a cryptic species related to the pan-Amazonian frog Allobates femoralis (Boulenger 1883) (Anura: Aromobatidae). Zootaxa, 2406, 1–28.
Sites, J.W. & Marshall, J.C. (2004) Operational criteria for delimiting species. Annual Review of Ecology Evolution and Systematics, 35, 199–227.
SPSS Inc. (2004) Statistical Package for the Social Sciences, version 13.0 for Windows, Chicago, E.U.A.
Stamatakis, A. (2006) RAxML-VI-HPC: Maximum likelihood-based phylogenetic analyses with thousands of taxa and mixed models. Bioinformatics, 22, 2688–2690.
Wiens, J.J. (2008) Systematics and herpetology in the age of genomics. Bioscience, 58, 297–307.
Wiley, E.O. (1978) The evolutionary species concept reconsidered. Systematic Zoology, 27, 17–26.
Yasuní-ITT. (2009) Yasuní-ITT, una iniciativa para cambiar la historia. [Web application]. Gobierno Nacional de la República del Ecuador. Available at: http://www.yasuni-itt.gov.ec (Accessed 25 February 2010).
Zwickl, D. (2006) Genetic algorithm approaches for the phylogenetic analysis of large biological sequence datasets under the maximum likelihood criterion. Ph.D. dissertation, The University of Texas at Austin. [Web application] Available at: http://www.bio.utexas.edu/faculty/antisense/garli/Garli.html. (Accessed January 2010).