Skip to main content Skip to main navigation menu Skip to site footer
Type: Article
Published: 2018-06-20
Page range: 79–104
Abstract views: 149
PDF downloaded: 5

When the unknown lives next door: a study of central Amazonian anurofauna

Laboratório de Evolução e Genética Animal (LEGAL), Departamento de Genética, Instituto de Ciências Biológicas, Universidade Federal do Amazonas (UFAM), Av. General Rodrigo Otávio Jordão Ramos, 6200, 69077-000 Manaus, AM, Brazi
Laboratório de Zoologia, Departamento de Biologia, Instituto de Ciências Biológicas, Universidade Federal do Amazonas (UFAM), Av. General Rodrigo Otávio Jordão Ramos, 6200, 69077-000 Manaus, AM, Brazil Programa de Pós-Graduação em Zoologia, Instituto de Ciências Biológicas, Universidade Federal do Amazonas (UFAM), Av. General Rodrigo Otávio Jordão Ramos, 6200, 69077-000 Manaus, AM, Brazil
Laboratório de Evolução e Genética Animal (LEGAL), Departamento de Genética, Instituto de Ciências Biológicas, Universidade Federal do Amazonas (UFAM), Av. General Rodrigo Otávio Jordão Ramos, 6200, 69077-000 Manaus, AM, Brazil Programa de Pós-Graduação em Zoologia, Instituto de Ciências Biológicas, Universidade Federal do Amazonas (UFAM), Av. General Rodrigo Otávio Jordão Ramos, 6200, 69077-000 Manaus, AM, Brazil
Laboratório de Evolução e Genética Animal (LEGAL), Departamento de Genética, Instituto de Ciências Biológicas, Universidade Federal do Amazonas (UFAM), Av. General Rodrigo Otávio Jordão Ramos, 6200, 69077-000 Manaus, AM, Brazil Programa de Pós-Graduação em Zoologia, Instituto de Ciências Biológicas, Universidade Federal do Amazonas (UFAM), Av. General Rodrigo Otávio Jordão Ramos, 6200, 69077-000 Manaus, AM, Brazil
Laboratório de Evolução e Genética Animal (LEGAL), Departamento de Genética, Instituto de Ciências Biológicas, Universidade Federal do Amazonas (UFAM), Av. General Rodrigo Otávio Jordão Ramos, 6200, 69077-000 Manaus, AM, Brazil Programa de Pós-Graduação em Zoologia, Instituto de Ciências Biológicas, Universidade Federal do Amazonas (UFAM), Av. General Rodrigo Otávio Jordão Ramos, 6200, 69077-000 Manaus, AM, Brazil
Reptilia 16S rRNA gene Anura cryptic species Amazonia

Abstract

The number of species of anurans in the Amazon is highly underestimated with new studies reporting the discovery of a large number of species every year. This advance in the discovery of biodiversity is due to the use of molecular tools, especially 16S rRNA gene barcoding, which is used to identify species and discover cryptic lineages. Few anurans of the central Amazon have molecular sequence data available in public databases, which contrasts with the considerable species richness of this biome. The aim of the present study was to test for the presence of cryptic species using the mPTP delimitation algorism. We morphologically identified 26 species, of which 23 were confirmed molecularly with the remaining three species identified as other congeneric species, since sequences with the same epithet do not exist in GenBank. Of these 23 species, nine contained one lineage restricted to central Amazon. This represents an underestimate of 39% in the taxonomic diversity in our sample. This is particularly surprising given that our sampling sites are among the best-studied regions of the central Amazon.

 

References

  1. Betancourth-Cundar, M., Lima, A.P., Hӧdl, W. & Amézquita, A. (2016) Decoupled evolution between senders and receivers in the Neotropical Allobates femoralis frog complex. PLOS ONE, 11, e0155929.
    https://doi.org/10.1371/journal.pone.0155929

    Biju, S.D. & Bossuyt, F. (2003) New frog family from India reveals an ancient biogeographical link with the Seychelles. Nature, 425, 711.
    https://doi.org/10.1038/nature02019

    Camargo, A., de Sá, R.O. & Heyer, W.R. (2006) Phylogenetic analyses of mtDNA sequences reveal three cryptic lineages in the widespread neotropical frog Leptodactylus fuscus (Schneider, 1799) (Anura, Leptodactylidae). Biological Journal of the Linnean Society, 87 (2), 325–341.
    https://doi.org/10.1111/j.1095-8312.2006.00581.x

    Caminer, M.A. & Ron, S.R. (2014) Systematics of tree frogs of the Hypsiboas calcaratus and Hypsiboas fasciatus species complex (Anura, Hylidae) with the description of four new species. ZooKeys, 68, 1–68.
    https://doi.org/10.3897/zookeys.370.6291

    Canedo, C. & Haddad, C.F.B. (2012) Phylogenetic relationships within anuran clade Terrarana, with emphasis on the placement of Brazilian Atlantic rainforest frogs genus Ischnocnema (Anura: Brachycephalidae). Molecular Phylogenetics and Evolution, 65 (2), 610–620.
    https://doi.org/10.1016/j.ympev.2012.07.016

    Casiraghi, M., Labra, M., Ferri, E., Galimberti, A. & de Mattia, F. (2010) DNA barcoding: A six-question tour to improve users’ awareness about the method. Briefings in Bioinformatics, 11 (4), 440–453.
    https://doi.org/10.1093/bib/bbq003

    Crawford, A.J., Lips, K.R. & Bermingham, E. (2010) Epidemic disease decimates amphibian abundance, species diversity, and evolutionary history in the highlands of central Panama. Proceedings of the National Academy of Sciences, 107 (31), 13777–13782.
    https://doi.org/10.1073/pnas.0914115107

    Cruz-Piedrahita, C., Navas, C.A. & Crawford, A.J. (2017) Life on the edge: a comparative study of ecophysiological adaptations of frogs to tropical semiarid environments. Physiological and Biochemical Zoology, 91 (1), 740–756.
    https://doi.org/10.1086/695705

    Darriba, D., Taboada, G.L., Doallo, R. & Posada, D. (2012) jModelTest 2: more models, new heuristics and parallel computing. Nature Methods, 9, 772.
    https://doi.org/10.1038/nmeth.2109

    Darst, C.R. & Cannatella, D.C. (2004) Novel relationships among hyloid frogs inferred from 12S and 16S mitochondrial DNA sequences. Molecular Phylogenetics and Evolution, 31 (2), 462–475.
    https://doi.org/10.1016/j.ympev.2003.09.003

    de Sá, R.O., Heyer, W.R. & Camargo, A. (2005a) A Phylogenetic analysis of Vanzolinius Heyer, 1974 (Amphibia, Anura, Leptodactylidae): Taxonomic and life history implications. Arquivos do Museu Nacional Rio de Janeiro, 63 (4), 707–726.

    de Sá, R.O., Heyer, W.R. & Camargo, A. (2005b) Are Leptodactylus didymus and L. mystaceus phylogenetically sibling species (Amphibia, Anura, Leptodactylidae)? Herpetologia Petropolitana, 2005, 90–92.

    de Sá, R.O., Streicher, J.W., Sekonyela, R., Forlani, M.C., Loader, S.P., Greenbaum, E., Richards, S. & Haddad, C.F.B. (2012) Molecular phylogeny of microhylid frogs (Anura: Microhylidae) with emphasis on relationships among New World genera. BMC Evolutionary Biology, 12, 241.
    https://doi.org/10.1186/1471-2148-12-241

    de Sá, R.O., Grant, T., Camargo, A., Heyer, W.R., Ponssa, M.L. & Stanley, E. (2014) Systematics of the Neotropical genus Leptodactylus Fitzinger, 1826 (Anura: Leptodactylidae): phylogeny, the relevance of non-molecular evidence, and species accounts. South American Journal of Herpetology, 9, 1–100. [S1–S100]
    https://doi.org/10.2994/SAJH-D-13-00022.1

    Evans, B.J., Kelley, D.B., Tinsley, R.C., Melnick, D.J. & Cannatella, D.C. (2004) A mitochondrial DNA phylogeny of African clawed frogs: phylogeography and implications for polyploid evolution. Molecular Phylogenetics and Evolution, 33 (1), 197–213.
    https://doi.org/10.1016/j.ympev.2004.04.018

    Faivovich, J., García, P.C.A., Ananias, F., Lanari, L., Basso, N.G. & Wheeler, W.C. (2004) A molecular perspective on the phylogeny of the Hyla pulchella species group (Anura, Hylidae). Molecular Phylogenetics and Evolution, 32, 938–950.
    https://doi.org/10.1016/j.ympev.2004.03.008

    Faivovich, J., Haddad, C.F.B., Garcia, P.C.A., Frost, D.R., Campbell, J.A. & Wheeler, W.C. (2005) Systematic review of the frog family Hylidae, with special reference to the Hylinae: Phylogenetic analysis and taxonomic revision. Bulletin of the American Museum of Natural History, 294, 1–240.
    https://doi.org/10.5531/sd.sp.12

    Faivovich, J., Haddad, C.F.B., Baêta, D., Jungfer, K.-H., Álvares, G.F.R., Brandão, R.A., Sheil, C., Barrientos, L.S., Barrio-Amorós, C.L., Cruz, C.A.G. & Wheeler, W.C. (2010) The phylogenetic relationships of the charismatic poster frogs, Phyllomedusinae (Anura, Hylidae). Cladistics, 26, 227–261.

    Fouquet, A., Gilles, A., Vences, M., Marty, C., Blanc, M. & Gemmell, N.J. (2007a) Underestimation of species richness in Neotropical frogs revealed by mtDNA analyses. PLoS ONE, 2, e1109.
    https://doi.org/10.1371/journal.pone.0001109

    Fouquet, A., Vences, M., Salducci, M. D., Meyer, A., Marty, C., Blanc, M. & Gilles, A. (2007b) Revealing cryptic diversity using molecular phylogenetics and phylogeography in frogs of the Scinax ruber and Rhinella margaritifera species groups. Molecular Phylogenetics and Evolution, 43 (2), 567–582.
    https://doi.org/10.1016/j.ympev.2006.12.006

    Fouquet, A., Noonan, B.P., Rodrigues, M.T., Pech, N., Gilles, A. & Gemmell, N.J. (2012) Multiple quaternary refugia in the eastern Guiana Shield revealed by comparative phylogeography of 12 frog species. Systematic Biology, 61 (3), 461.
    https://doi.org/10.1093/sysbio/syr130

    Fouquet, A., Pineau, K., Rodrigues, M.T., Mailles, J., Schneider, J.B., Ernst, R. & Dewynter, M. (2013a) Endemic or exotic: the phylogenetic position of the Martinique Volcano Frog Allobates chalcopis (Anura: Dendrobatidae) sheds light on its origin and challenges current conservation strategies. Systematics and Biodiversity, 11 (1), 87–101.
    https://doi.org/10.1080/14772000.2013.764944

    Fouquet, A., Blotto, B.L., Maronna, M.M., Verdade, V.K., Junc, F.A., de S, R. & Rodrigues, M.T. (2013b) Unexpected phylogenetic positions of the genera Rupirana and Crossodactylodes reveal insights into the biogeography and reproductive evolution of leptodactylid frogs. Molecular Phylogenetics and Evolution, 67 (2), 445–457.
    https://doi.org/10.1016/j.ympev.2013.02.009

    Fouquet, A., Courtois, E.A., Baudain, D., Lima, J.D., Souza, S.M., Noonan, B.P. & Rodrigues, M.T. (2015a) The trans-riverine genetic structure of 28 Amazonian frog species is dependent on life history. Journal of Tropical Ecology, 31 (4), 361–373.
    https://doi.org/10.1017/S0266467415000206

    Fouquet, A., Orrico, V.G., Ernst, R., Blanc, M., Martinez, Q., Vacher, J.P., Rodrigues, M.T., Ouboter, P., Jairam, R. & Ron, S. (2015b) A new Dendropsophus Fitzinger, 1843 (Anura: Hylidae) of the parviceps group from the lowlands of the Guiana Shield. Zootaxa, 4052 (1), 39–64.
    https://doi.org/10.11646/zootaxa.4052.1.2

    Fouquet, A., Martinez, Q., Zeidler, L., Courtois, E.A., Gaucher, P., Blanc, M., Lima, J.D., Souza, S.M., Rodrigues, M.T. & Kok, P.J.R. (2016) Cryptic diversity in the Hypsiboas semilineatus species group (Amphibia, Anura) with the description of a new species from the eastern Guiana Shield. Zootaxa, 4084 (1), 79–104.
    https://doi.org/10.11646/zootaxa.4084.1.3

    Frost, D.R., Grant, T., Faivovich, J., Bain, R.H., Haas, A., Haddad, C.F.B., De Sa, R.O., Channing, A., Wilkinson, M., Donnellan, S.C., Raxworthy, C.J., Campbell, J.A., Blotto, B.L., Moler, P., Drewes, R.C., Nussbaum, R.A., Lynch, J.D., Green, D.M. & Wheeler, W.C. (2006) The amphibian tree of life. Bulletin of the American Museum of Natural History, 297, 1–370.
    https://doi.org/10.5531/sd.sp.13

    Frost, D.R. (2017) Amphibian Species of the World: an Online Reference. Version 6.0. American Museum of Natural History, New York, USA. Available from: http://research.amnh.org/herpetology/amphibia/ (accessed 20 July 2017)

    Funk, W.C., Caminer, M. & Ron, S.R. (2012) High levels of cryptic species diversity uncovered in Amazonian frogs. Proceedings of the Royal Society B: Biological Sciences, 279 (1734), 1806–1814.
    https://doi.org/10.1098/rspb.2011.1653

    Gazoni, T., Haddad, C.F.B., Narimatsu, H., Cabral-de-Mello, D.C., Lyra, M.L. & Parise-Maltempi, P.P. (2018) More sex chromosomes than autosomes in the Amazonian frog Leptodactylus pentadactylus. Chromosoma. [published online]
    https://doi.org/10.1007/s00412-018-0663-z

    Grant, T., Frost, D.R., Caldwell, J.P., Gagliardo, R.O.N., 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.
    https://doi.org/10.5531/sd.sp.14

    Guarnizo, C.E., Paz, A., Muñoz-Ortiz, A., Flechas, S.V., Méndez-Narváez, J. & Crawford, A.J. (2015) DNA barcoding survey of anurans across the eastern cordillera of Colombia and the impact of the Andes on cryptic diversity. PLoS ONE, 10, e0127312.
    https://doi.org/10.1371/journal.pone.0127312

    Hebert, P.D.N., Cywinska, A., Ball, S.L. & de Waard, J.R. (2003) Biological identifications through DNA barcodes. Proceedings of the Royal Society of London B: Biological Sciences, 270 (1512), 313–321.
    https://doi.org/10.1098/rspb.2002.2218

    Heinicke, M.P., Duellman, W.E. & Hedges, S.B. (2007) Major Caribbean and Central American frog faunas originated by ancient oceanic dispersal. Proceedings of the National Academy of Sciences of USA, 104 (24), 10092–10097.

    Hero, J.M. (1990) An illustrated key to tadpoles occurring in the central Amazon rainforest, Manaus, Amazonas, Brasil. Amazoniana, 11, 201–262.

    Heyer, W.R., de Sá, R.O. & Rettig, A. (2005) Sibling species, advertisement calls, and reproductive isolation in frogs of the Leptodactylus pentadactylus species cluster (Amphibia, Leptodactylidae). Herpetologia Petropolitana, 2003, 35–39.

    Heyer, W.R. & de Sá, R.O. (2011) Variation, systematics, and relationships of the Leptodactylus bolivianus complex (Amphibia: Anura: Leptodactylidae). Smithsonian Contributions to Zoology, 635, 1–58.
    https://doi.org/10.5479/si.00810282.635.1

    Jansen, M., Bloch, R., Schulze, A. & Pfenninger, M. (2011) Integrative inventory of Bolivia’s lowland anurans reveals hidden diversity. Zoologica Scripta, 40 (6), 567–583.
    https://doi.org/10.1111/j.1463-6409.2011.00498.x

    Jungfer, K.H., Faivovich, J., Padial, J.M., Castroviejo-Fisher, S., Lyra, M.M., Berneck, B.V. M., Iglesias, P.P., Kok, P.J.R., MacCulloch, R.D., Rodrigues, M.T., Verdade, V.K.,Torres Gastello, C.P., Chaparro, J.C., Valdujo, P.H., Reichle, S., Moravec, J., Gvoždík, V., Gagliardi-Urrutia, G., Ernst, R., De la Riva, I., Means, D.B., Lima, A.P., Señaris, J.C., Wheeler, W.C. & Haddad, C.F.B. (2013) Systematics of spiny-backed treefrogs (Hylidae: Osteocephalus): an Amazonian puzzle. Zoologica Scripta, 42 (4), 351–380.
    https://doi.org/10.1111/zsc.12015

    Kapli, P., Lutteropp, S., Zhang, J., Kobert, K., Pavlidis, P., Stamatakis, A. & Flouri, T. (2017) Multi-rate Poisson tree processes for single-locus species delimitation under maximum likelihood and Markov chain Monte Carlo. Bioinformatics, 33 (11), 1630–1638.
    https://doi.org/10.1093/bioinformatics/btx025

    Kok, P.J.R., MacCulloch, R.D., Means, D.B., Roelants, K., Van Bocxlaer, I. & Bossuyt, F. (2012) Low genetic diversity in tepui summit vertebrates. Current Biology, 22 (15), R589–R590.
    https://doi.org/10.1016/j.cub.2012.06.034

    Kok. P.J.R., Sebastian, R., MacCulloch, R.D., Amy, L., Raheleh, D., Fablen, A. & Bruce, M. (2017) Historical biogeography of the palaeoendemic toad genus Oreophrynella (Amphibia: Bufonidae) sheds a new light on the origin of the Pantepui endemic terrestrial biota. Journal of Biogeography, 45 (1), 26–36.
    https://doi.org/10.1111/jbi.13093

    Kurabayashi, A., Matsui, M., Belabut, D.M., Yong, H.-S., Ahmad, N., Sudin, A., Kuramoto, M., Hamidy, A. & Sumida, M. (2011) From Antarctica or Asia? New colonization scenario for Australian-New Guinean narrow mouth toads suggested from the findings on a mysterious genus Gastrophrynoides. BMC Evolutionary Biology, 11, 175.
    https://doi.org/10.1186/1471-2148-11-175

    Larkin, M.A., Blackshields, G., Brown, N.P., Chenna, R., McGettigan, P.A., McWilliam, H., Valentin, F., Wallace, I.M., Wilm, A., Lopez, R., Thompson, J.D., Gibson, T.J. & Higgins, D.G. (2007) Clustal W and Clustal X version 2.0. Bioinformatics, 23 (21), 2947–2948.
    https://doi.org/10.1093/bioinformatics/btm404

    Lewinsohn, T.M. & Prado, P.I. (2005) How many species are there in Brazil? Conservation Biology, 19 (3), 619–624.

    Lyra, M.L., Haddad, C.F.B. & Azeredo-Espin, A.M.L. (2016) Meeting the challenge of DNA barcoding Neotropical amphibians: polymerase chain reaction optimization and new COI primers. Molecular Ecology Resources, 17 (5), 966–980.
    https://doi.org/10.1111/1755-0998.12648

    Machado, D.J., Lyra, M.L. & Grant, T. (2016) Mitogenome assembly from genomic multiplex libraries: comparison of strategies and novel mitogenomes for five species of frogs. Molecular Ecology Resources, 16 (3), 686–693.
    https://doi.org/10.1111/1755-0998.12492

    Maciel, N.M., Collevatti, R.G., Colli, G.R. & Schwartz, E.F. (2010) Late Miocene diversification and phylogenetic relationships of the huge toads in the Rhinella marina (Linnaeus, 1758) species group (Anura: Bufonidae). Molecular Phylogenetic and Evolution, 57 (2), 787–797.
    https://doi.org/10.1111/1755-0998.12492

    Moravec, J. (2009) Dendropsophus reichlei. In: IUCN 2014. IUCN Red List of Threatened Species. Version 2014.1. Available from: http://www.iucnredlist.org (accessed 1 July 2014)

    Moravec, J., Aparicio, J., Guerrero-Reinhard, M., Calderón, G., Jungfer, K.H. & Gvoždík, V. (2009) A new species of Osteocephalus (Anura: Hylidae) from Amazonian Bolivia: First evidence of tree frog breeding in fruit capsules of the Brazil nut tree. Zootaxa, 54, 37–54.
    https://doi.org/10.5281/zenodo.189932

    Moravec, J, Gagliardi-Urrutia, G. & Gvoždík, V. (2016) Amphibians and reptiles recorded in the conservation area Imiría in the Ucayali region in Peru. Acta Societatis Zoologicae Bohemicae, 80, 317–341.

    Muñoz-Ortiz, A., Velásquez-Álvarez, .A., Guarnizo, C.E. & Crawford, A.J. (2015) Of peaks and valleys: Testing the roles of orogeny and habitat heterogeneity in driving allopatry in mid-elevation frogs (Aromobatidae: Rheobates) of the northern Andes. Journal of Biogeography, 42 (1), 193–205.
    https://doi.org/10.1111/jbi.12409

    Oliveira, E.A. De, Rodrigues, L.R., Kaefer, I.L., Pinto, K.C. & Hernández-Ruz, E.J. (2017) A new species of Pristimantis from eastern Brazilian Amazonia (Anura, Craugastoridae). ZooKeys, 687, 101–129.
    https://doi.org/10.3897/zookeys.687.13221

    Padial, J.M. & De La Riva, I. (2009) Integrative taxonomy reveals cryptic Amazonian species of Pristimantis (Anura: Strabomantidae). Zoological Journal of the Linnean Society, 155 (1), 97–122.
    https://doi.org/10.1111/j.1096-3642.2008.00424.x

    Padial, J.M., Castroviejo-Fisher, S., Köhler, J., Vil, C., Chaparro, J.C. & De La Riva, I. (2009) Deciphering the products of evolution at the species level: The need for an integrative taxonomy. Zoologica Scripta, 38 (4), 431–447.
    https://doi.org/10.1111/j.1463-6409.2008.00381.x

    Palumbi, S.R. (1996) Nucleic acids II: the polymerase chain reaction. In: Hillis, D.M., Moritz, C. & Mable, B.K. (Eds.), Molecular Systematics. Sinauer & Associates Inc., Sunderland, Massachusetts, pp. 205–247.

    Peloso, P.L.V., Pellegrino, K.C.M., Rodrigues, M.T. & Avila-Pires, T.C.S. (2011). Description and phylogenetic relationships of a new genus and species of lizard (Squamata, Gymnophthalmidae) from the amazonian rainforest of northern Brazil. American Museum Novitates, 3713, 1–24.
    https://doi.org/10.1206/3713.2

    Peloso, P.L.V., Sturaro, M.J., Forlani, M.C., Gaucher, P., Motta, A.P. & Wheeler, W.C. (2014) Phylogeny, taxonomic revision, and character evolution of the genera Chiasmocleis and Syncope (Anura, Microhylidae) in Amazonia, with descriptions of three new species. Bulletin of the American Museum of Natural History, 386, 1–112.
    https://doi.org/10.5531/sd.herp.2

    Peloso, P.L.V., Frost, D.R., Richards, S.J., Rodrigues, M.T., Donnellan, S., Matsui, M., Raxworthy, C.J., Biju, S.D., Lemmon, E.M., Lemmon, A.R. & Wheeler, W.C. (2015) The impact of anchored phylogenomics and taxon sampling on phylogenetic inference in narrow-mouthed frogs (Anura, Microhylidae). Cladistics, 32 (2), 113–140.
    https://doi.org/10.1111/cla.12118

    Pereira, E.B., Collevatti, R.G., Kokubum, M.N. de C., Miranda, N.E. de O. & Maciel, N.M. (2015) Ancestral reconstruction of reproductive traits shows no tendency toward terrestriality in leptodactyline frogs. BMC Evolutionary Biology, 15, 91.
    https://doi.org/10.1186/s12862-015-0365-6

    Pramuk, J.B. (2006) Phylogeny of South American Bufo (Anura: Bufonidae) inferred from combined evidence. Biological Journal of the Linnean Society, 146 (3), 407–452.
    https://doi.org/10.1111/j.1096-3642.2006.00212.x

    Rambaut, A. & Drummond, A.J. (2007) Tracer version 1.5. available from http://tree.bio.ed.ac.uk/software/tracer/ (accessed 3 June 2016)

    Rivera-Correa, M., Lamadrid-Feris, F. & Crawford, A.J. (2016) A new small golden frog of the genus Pristimantis (Anura: Craugastoridae) from an Andean cloud forest of Colombia. Amphibia-Reptilia, 37 (2), 153–166.
    https://doi.org/10.1163/15685381-00003037

    Roberts, J.L., Brown, J.L., May, R. von, Arizabal, W., Schulte, R. & Summers, K. (2006) Genetic divergence and speciation in lowland and montane peruvian poison frogs. Molecular Phylogenetics and Evolution, 41 (1), 149–164.
    https://doi.org/10.1016/j.ympev.2006.05.005

    Roelants, K. & Bossuyt, F. (2005) Archaeobatrachian paraphyly and pangaean diversification of crown-group frogs. Systematic Biology, 54 (1), 111–126.
    https://doi.org/10.1080/10635150590905894

    Rojas, R.R., Carvalho, V.T., Gordo, M., Ávila, R.W., Farias, I.P. & Hrbek, T. (2014) A new species of Amazophrynella (Anura: Bufonidae) from the southwestern part of the Brazilian Guiana Shield. Zootaxa, 3753 (1), 79–95.
    https://doi.org/10.11646/zootaxa.3753.1.7

    Ronquist, F., Teslenko, M., Van Der Mark, P., Ayres, D.L., Darling, A., Höhna, S., Larget, B., Liu, L., Suchard, M.A. & Huelsenbeck, J.P. (2012) MrBayes 3.2: Efficient Bayesian phylogenetic inference and model choice across a large model space. Systematic Biology, 61 (3), 539–542.
    https://doi.org/10.1093/sysbio/sys029

    Saitou, N. & Nei M. (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution, 24 (4), 406–425.
    https://doi.org/10.1093/oxfordjournals.molbev.a040454

    Sambrook, J., Fritsch, E.F. & Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual. 2nd Edition. Cold Spring Harbor Laboratory Press, Cold Springs Harbor, New York, 1626 pp.

    Salducci, M.-D., Marty, C., Fouquet, A. & Gilles, A. (2005) Phylogenetic relationships and biodiversity in Hylids (Anura: Hylidae) from French Guiana. Comptes Rendus Biologies, 328 (10–11), 1009–1024.
    https://doi.org/10.1016/j.crvi.2005.07.005

    Salerno, P.E., Ron, S.R., Señaris, J.C., Rojas-Runjaic, F.J.M., Noonan, B.P. & Cannatella, D.C. (2012) Ancient tepui summits harbor young rather than old lineages of endemic frogs. Evolution, 66 (10), 3000–3013.
    https://doi.org/10.1111/j.1558-5646.2012.01666.x

    Santos, J.C. & Cannatella, D.C. (2011) Phenotypic integration emerges from aposematism and scale in poison frogs. Proceedings of the National Academy of Sciences, 108 (15), 6175–6180.
    https://doi.org/10.1073/pnas.1010952108

    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, e56.
    https://doi.org/10.1371/journal.pbio.1000056

    Schulze, A., Jansen, M. & Koöhler, G. (2015) Tadpole diversity of Bolivia’s lowland anuran communities: Molecular identification, morphological characterisation, and ecological assignment. Zootaxa, 4016 (1), 1–111.
    https://doi.org/10.11646/zootaxa.4016.1.1

    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.

    Simões, P.I. & Lima, A.P. (2012) The tadpole of Allobates sumtuosus (Morales, ‘2000’ 2002)(Anura: Aromobatidae) from its type locality at Reserva Biológica do Rio Trombetas, Pará, Brazil. Zootaxa, 3499, 86–88.

    Simões, P.I., Kaefer, I.L., Farias, I.P. & Lima, A.P. (2013) An integrative appraisal of the diagnosis and distribution of Allobates sumtuosus (Morales, 2002) (Anura, Aromobatidae). Zootaxa, 3746 (3), 401–421.
    https://doi.org/10.11646/zootaxa.3746.3.1

    Simões, P.I., Stow, A., Hödl, W., Amézquita, A., Farias, I.P. & Lima, A.P. (2014) The value of including intraspecific measures of biodiversity in environmental impact surveys is highlighted by the Amazonian brilliant-thighed frog (Allobates femoralis). Tropical Conservation Science, 7, 811–828.
    https://doi.org/10.1177/194008291400700416

    Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M. & Kumar, S. (2011) MEGA5: Molecular Evolutionary Genetics Analysis using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony Methods. Molecular Biology and Evolution, 28 (10), 2731–2739.
    https://doi.org/10.1093/molbev/msr121.

    Thompson, C. (2010) Amazon alive! A decade of discovery: 1999–2009. WWF, 62 pp. Available from: http://www.wwf.org.br/informacoes/bliblioteca/publications_in_english/?uNewsID=26346 (accessed 28 February 2015)

    Tsuji-Nishikido, B.M., Kaefer, I.L., de Freitas, F.C., Menin, M. & Lima, A.P. (2012) Significant but not diagnostic: differentiation through morphology and calls in the Amazonian frogs Allobates nidicola and A. masniger. Herpetological Journal, 22 (2), 105–114.

    Vacher, J.P., Kok, P.J.R., Rodrigues, M.T., Lima, J.D., Lorenzini, A., Martinez, Q., Fallet, M., Courtois, E.A., Blanc, M., Gaucher, P., Dewynter, M., Jairam, R., Ouboter, P., Thébaud, C. & Fouquet, A. (2017) Cryptic diversity in Amazonian frogs: Integrative taxonomy of the genus Anomaloglossus (Amphibia: Anura: Aromobatidae) reveals a unique case of diversification within the Guiana Shield. Molecular Phylogenetics and Evolution, 112, 158–173.

    https://doi.org/10.1016/j.ympev.2017.04.017

    Vences, M., Kosuch, J., Lotters, S., Widmer, A., Jungfer, K.H., Kohler, J. & Veith, M. (2000) Phylogeny and classification of poison frogs (Amphibia: Dendrobatidae), based on mitochondrial 16S and 12S ribosomal RNA gene sequences. Molecular Phylogenetics and Evolution, 15 (1), 34–40.
    https://doi.org/10.1006/mpev.1999.0738

    Vences, M., Kosuch, J., Boistel, R., Haddad, C.F.B., La Marca, E., Lötters, S. & Veith, M. (2003) Convergent evolution of aposematic coloration in Neotropical poison frogs: a molecular phylogenetic perspective. Organisms Diversity & Evolution, 3 (3), 215–226.
    https://doi.org/10.1078/1439-6092-00076

    Vences, M., Thomas, M., van der Meijden, A., Chiari, Y. & Vieites, D. (2005) Comparative performance of the 16S rRNA gene in DNA barcoding of amphibians. Frontiers in Zoology, 2 (5), 1–12.

    https://doi.org/10.1186/1742-9994-2-5

    Vieites, D.R., Wollenberg, K.C. & Vences, M. (2012) Not all little brown frogs are the same: A new species of secretive and cryptic Gephyromantis (Anura: Mantellidae) from Madagascar. Zootaxa, 46, 34–46.
    https://doi.org/10.5281/zenodo.281463

    Yanek, K., Heyer, W.R. & De S, R.O. (2006) Genetic resolution of the enigmatic lesser Antillean distribution of the frog Leptodactylus validus (Anura, Leptodactylidae). South American Journal of Herpetology, 1 (3), 192–201.
    https://doi.org/10.2994/1808-9798(2006)1[192:grotel]2.0.co;2

    Zhang, J., Kapli, P., Pavlidis, P. & Stamatakis, A. (2013a) A general species delimitation method with applications to phylogenetic placements. Bioinformatics, 29 (22), 2869–2876.
    https://doi.org/10.1093/bioinformatics/btt499

    Zhang, P. & Wake, M.H. (2009) A mitogenomic perspective on the phylogeny and biogeography of living caecilians (Amphibia: Gymnophiona). Molecular Phylogenetics and Evolution, 53 (2), 479–491.
    https://doi.org/10.1016/j.ympev.2009.06.018

    Zhang, P., Liang, D., Mao, R.-L., Hillis, D.M., Wake, D.B. & Cannatella, D.C. (2013b) Efficient sequencing of Anuran mtDNAs and a mitogenomic exploration of the phylogeny and evolution of frogs. Molecular Biology and Evolution, 30 (8), 1899–1915.
    https://doi.org/10.1093/molbev/mst091