Skip to main content Skip to main navigation menu Skip to site footer
Type: Article
Published: 2023-06-02
Page range: 301-336
Abstract views: 1299
PDF downloaded: 519

A new species of barred frog, Mixophyes (Anura: Myobatrachidae) from south-eastern Australia identified by molecular genetic analyses

School of Environmental and Life Sciences, University of Newcastle, University Drive, Callaghan 2308, Australia
South Australian Museum, North Terrace, Adelaide 5000, Australia. School of Biological Sciences, University of Adelaide, North Terrace, Adelaide 5005, Australia.
South Australian Museum, North Terrace, Adelaide 5000, Australia
Queensland Parks and Wildlife Service and Partnerships, Department of Environment and Science, PO Box 64, Bellbowrie, Qld 4070 and Honorary Research Fellow, Biodiversity, Queensland Museum, PO Box 3300, South Brisbane, Qld 4101
South Australian Museum, North Terrace, Adelaide 5000, Australia
IUCN threat category morphology mtDNA nuclear DNA sequences species boundaries Amphibia

Abstract

Mixophyes are large ground-dwelling myobatrachid frogs from eastern Australia and New Guinea. Several of the species found in mid-eastern and south-eastern Australia are listed as threatened, due largely to declines presumably caused by the amphibian disease chytridiomycosis. Given the wide distribution of several of these species and that their distributions cross well-known biogeographic boundaries that often correspond to deep genetic breaks or species boundaries among closely related vertebrates, we undertook a molecular genetic assessment of population structure across the range of each species to determine the presence of undescribed species. Of the four species of Mixophyes subject to molecular population genetic analyses, one, the Stuttering Frog (Mixophyes balbus), showed a level of diversity consistent with the presence of two species. Morphometric, meristic and bioacoustic analyses corroborate these distinctions, and a new species is described for the populations south of the Macleay River valley in mid-eastern New South Wales to east Gippsland in Victoria. Applying the IUCN Red List threat criteria the new species meets the conservation status assessment criteria for Endangered 2B1a,b because its extent of occupancy and area of occupancy are below the threshold value and it has declined and disappeared from the southern two thirds of its distribution over the past 30 years.

References

  1. Anstis, M. (2013) Tadpoles and frogs of Australia. New Holland Publishers, Sydney, 829 pp.
    Anstis, M., Price, L.C., Roberts, J.D., Catalano, S., Doughty, P., Hines, H.B. & Donnellan, S.C. (2016) Revision of the Australian water holding frog (Cyclorana platycephala, Anura: Hylidae), with a description of a new species and subspecies. Zootaxa, 4126 (4), 451–479.
    https://doi.org/10.11646/zootaxa.4126.4.1
    Barker, J. Grigg, G.C. & Tyler, M.J. (1995) A Field Guide to Australian Frogs. Surrey Beatty & Sons, Sydney, 407 pp.
    Banks, C.H., Traher, R.U. & Hobbs, R.A. (2014) Captive management and breeding of the stuttering frog (Mixophyes balbus) at Melbourne Zoo. Herpetological Review, 45, 43–9.
    Beeman, K. (1998) Digital signal analysis, editing, and synthesis. In: Hopp, S.L., Owren, M.J. & Evans, C.S. (Eds.), Animal acoustic communication Springer, Berlin and Heidelberg, pp. 59–103.
    Berger, L., Speare, R., Daszak, P., Green, D.E., Cunningham, A.A., Goggin, C.L., Slocombe, R., Ragan, M.A., Hyatt, A.D., McDonald, K.R. & Hines, H.B. (1998) Chytridiomycosis causes amphibian mortality associated with population declines in the rain forests of Australia and Central America. Proceedings of the National Academy of Sciences USA, 95, 9031–9036.
    https://doi.org/10.1073/pnas.95.15.9031
    Chan, K.O., Grismer, L.L. (2022) Correcting for body size variation in morphometric analysis. Zootaxa, 5124, 471–482.
    https://doi.org/10.11646/zootaxa.5124.4.4
    Corben, C.J. & Ingram, G.J. (1987) A new barred river frog (Myobatrachidae: Mixophyes). Memoirs of the Queensland Museum, 25, 233–237.
    Cogger, H. (2014) Reptiles and amphibians of Australia. CSIRO Publishing, Collingwood, 1056 pp.
    Daly, G. (1998) Review of the status and assessment of the habitat of the Stuttering Frog Mixophyes balbus (Anura: Myobatrachidae) on the south coast of New South Wales. Herpetofauna, 28, 2–11.
    Daly, G. & Craven, P. (2011) The disappearance of the Stuttering Frog Mixophyes balbus at Macquarie Pass National Park, New South Wales. Australian Zoologist, 35, 846–852.
    Donnellan, S.C. & Mahony, M.J. (2004) Allozyme, chromosomal and morphological variability in the Litoria lesueuri species group (Anura: Hylidae), including a description of a new species. Australian Journal of Zoology, 52, 1–28.
    https://doi.org/10.1071/ZO02069
    Donnellan, S.C., McGuigan, K., Knowles, R., Mahony, M. & Moritz, C. (1999) Genetic evidence for species boundaries in frogs of the Litoria citropa species group (Anura: Hylidae). Australian Journal of Zoology, 47, 275–293.
    https://doi.org/10.1071/ZO99013
    Donnellan, S.C., Mahony, M. & Davies, M. (1990) A new species of Mixophyes and first record of the genus in New Guinea. Herpetologica, 46, 266–274.
    https://www.jstor.org/stable/3892969
    Donnellan, S.C., Catalano, S., Pederson, S., Mitchell, K., Suhendran, A., Price, L.C., Doughty, P. & Richards, S.J. (2021) Revision of the Litoria watjulumensis (Anura: Pelodryadidae) group from the Australian monsoonal tropics, including the resurrection of L. spaldingi. Zootaxa, 4933 (2), 211–240.
    https://doi.org/10.11646/zootaxa.4933.2.3
    Earl, D.A. (2012) Structure harvester: A website and program for visualizing structure output and implementing the Evanno method. Conservation Genetics Resources, 4, 359–361.
    https://doi.org/10.1007/s12686-011-9548-7
    Edgar, R.C. (2004) MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Research, 32, 1792–1797.
    https://doi.org/10.1093/nar/gkh340.
    Evanno, G., Regnaut, S. & Goudet, J. (2005) Detecting the number of clusters of individuals using the software structure: A simulation study. Molecular Ecology, 14, 2611–2620.
    https://doi.org/10.1111/j.1365-294X.2005.02553.x
    Georges, A, Gruber, B., Pauly, G., White, D., Adams, M., Young, M., Kilian, A., Zhang, X., Shaffer, H. & Unmack, P. (2018) Genome-wide SNP markers breathe new life into phylogeography and species delimitation for the problematic short-necked turtles (Chelidae: Emydura) of eastern Australia. Molecular Ecology, 27, 5195–5213.
    https://doi.org/10.1111/mec.14925
    Gerhardt, H.C. & Huber, F. (2002) Acoustic communication in insects and anurans: common problems and diverse solutions. University of Chicago Press, Chicago, Illinois, 531 pp.
    Gerhardt, H.C., Humfeld, S.C. & Marshall, V.T. (2007) Temporal order and the evolution of complex acoustic signals. Proceedings of the Royal Society B: Biological Sciences, 274, 1789–1794.
    https://doi.org/10.1098/rspb.2007.0451
    Geyle, H.M., Hoskin, C.J., Bower, D.S., Catullo, R., Clulow, S., Driessen, M., Daniels, K., Garnett, S.T., Gilbert, D., Heard, G.W., Hero, J-M., Hines, H.B., Hoffmann, E.P., Hollis, G., Hunter, D.A., Lemckert, F., Mahony, M., Marantelli, G., McDonald, K.R., Mitchell, N.J., Newell, D., Roberts, J.D., Scheele, B.C., Scroggie, M.P., Vanderduys, E., Wassens, S., West, M., Woinarski, J.C.Z & Gillespie, G.R. (2021) Red hot frogs: identifying the Australian frogs most at risk of extinction. Pacific Conservation Biology, 28, 211–223.
    https://doi.org/10.1071/PC21019
    Gillespie, G.R. (2011) Survey for the southern barred frog Mixophyes balbus in Victoria. Unpublished report to the Department of Sustainability and Environment, Orbost, Victoria.
    Gillespie, G.R. & Hines, H.B. (1999) Status of temperate riverine frogs in south-eastern Australia. In: Campbell, A. (Eds.), Declines and Disappearances of Australian Frogs. Environment Australia, Canberra, pp. 109–130.
    Gillespie, G.R., Kum, K.C., De Angelis, D.A. & Jenner, B.D. (2014) Gone but not forgotten: The status of the Stuttering Frog Mixophyes balbus in Victoria. The Victorian Naturalist, 131, 64–71.
    Gillespie, G.R., Roberts, J.D., Hunter, D., Hoskin, C.J., Alford, R.A., Heard, G.W., Hines, H., Lemckert, F., Newell, D. & Scheele, B.C. (2020) Status and priority conservation actions for Australian frog species. Biological Conservation, 247, 108543.
    https://doi.org/10.1016/J.BIOCON.2020.108543
    Gruber, B., Unmack, P.J., Berry, O.F. & Georges, A. (2018) dartr: An R package to facilitate analysis of SNP data generated from reduced representation genome sequencing. Molecular Ecology Resources, 18, 691–699.
    https://doi.org/10.1111/1755-0998.12745
    Hero, J.M., Morrison, C., Gillespie, G., Roberts, J.D., Newell, D., Meyer, E., McDonald, K., Lemckert, F., Mahony, M., Osborne, W. & Hines, H. (2006) Overview of the conservation status of Australian frogs. Pacific Conservation Biology, 12, 313–320.
    https://doi.org/10.1071/PC060313
    Hoang, D.T., Chernomor, O., von Haeseler, A., Minh, B.Q. & Vinh, L.S. (2018) UFBoot2: Improving the ultrafast bootstrap approximation. Molecular Biology Evolution, 35, 518–522.
    https://doi.org/10.1093/molbev/msx281
    Hoskin, C.J. (2010) Breeding behaviour of the Barred Frog Mixophyes coggeri. Memoirs of the Queensland Museum-Nature, 55, 1–7.
    Hunter, D. & Gillespie, G.R. (2011) National Recovery Plan for the Stuttering Frog Mixophyes balbus. Department of Sustainability and Environment, Victoria. [unknown pagination]
    IUCN (2019) Guidelines for using the IUCN Red List categories and criteria. Version 13. IUCN, Gland, Switzerland. Available from: http://www.iucnredlist.org/documents/RedListGuidelines.pdf (accessed 17 April 2023)
    Jombart, T. (2008) adegenet: a R package for the multivariate analysis of genetic markers. Bioinformatics, 24, 1403–1405.
    https://doi.org/10.1093/bioinformatics/btn129
    Kalyaanamoorthy, S., Minh, B.Q., Wong, T.K.F., von Haeseler, A. & Jermiin, L.S. (2017) ModelFinder: fast model selection for accurate phylogenetic estimates. Nature Methods, 14, 587–589.
    https://doi.org/10.1038/nmeth.4285
    Kearse, M., Moir, R., Wilson, A., Stones-Havas, S., Cheung, M., Sturrock, S., Buxton, S., Cooper, A., Markowitz, S., Duran, C., Thierer, T., Ashton, B., Meintjes, P. & Drummond, A. (2012) Geneious Basic: An integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics Applications Note, 28, 1647–164.
    https://doi.org/10.1093/bioinformatics/bts199
    Kehlmaier, C., Zhang, X., Georges, A., Campbell, P.D., Thomson, S. & Fritz, U. (2019) Mitogenomics of historical type specimens of Australasian turtles: clarification of taxonomic confusion and old mitochondrial introgression. Science Reports, 9, 5841.
    https://doi.org/10.1038/s41598-019-42310-x
    Kilian, A., Wenzl, P., Huttner, E., Carling, J., Xia, L., Blois, H., Caig, V., Heller-Uszynska, K., Jaccoud, D., Hopper, C., Aschenbrenner-Kilian, M., Evers, M., Peng, K., Cayla, C., Hok, P. & Uszynski, G. (2012) Diversity arrays technology: A generic genome profiling technology on open platforms. Methods in Molecular Biology, 888, 67–89.
    https://doi.org/10.1007/978-1-61779-870-2
    Köhler, J., Jansen, M., Rodríguez, A., Kok, P.J.R., Toledo, L.F., Emmrich, M., Glaw, F., Haddad, C.F.B., Rödel, M-O. & Vences, M. (2017) The use of bioacoustics in anuran taxonomy: theory, terminology, methods and recommendations for best practice. Zootaxa, 4251 (1), 1–124.
    https://doi.org/10.11646/zootaxa.4251.1.1
    Kopelman, N.M., Mayzel, J., Jakobsson, M., Rosenberg, N.A. & Mayrose, I. (2015) CLUMPAK: a program for identifying clustering modes and packaging population structure inferences across K. Molecular Ecology Resources, 15, 1179–1191.
    https://doi.org/10.1111/1755-0998.12387
    Knowles, R., Thumm, K., Mahony, M., Hines, H., Newell, D. & Cunningham, M. (2015) Oviposition and egg mass morphology in barred frogs (Anura: Myobatrachidae: Mixophyes Gunther, 1864), its phylogenetic significance and implications for conservation management. Australian Zoologist, 37, 381–402.
    https://doi.org/10.7882/AZ.2014.040
    Knowles, R., Mahony, M., Armstrong, J. & Donnellan, S.C. (2004) Systematics of sphagnum frogs of the genus Philoria (Anura: Myobatrachidae) in eastern Australia. Records of the Australian Museum, 56, 57–74.
    https://doi.org/10.3853/j.0067-1975.56.2004.1391
    Kumar, S., Stecher, G. & Tamura, K. (2016) MEGA7: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Molecular Biology Evolution, 33, 1870–1874.
    https://doi.org/10.1093/molbev/msw054
    Leaché, A.D., Banbury, B.L., Felsenstein, J., de Oca, A.N. & Stamatakis, A. (2015) Short tree, long tree, right tree, wrong tree: new acquisition bias corrections for inferring SNP phylogenies. Systematic Biology, 64, 1032–1047.
    https://doi.org/10.1093/sysbio/syv053
    Legge, S., Woinarski, J.C.S., Scheele, B.C., Garnett, S.T., Lintermans, M., Nimmo, D.G., Whiterod, N.S., Southwell, D.S., Ehmke, G., Buchan, A., Gray, J., Metcalfe, D.J., Page, M., Rumpff, L., van Leeuwen, S., Williams, D., Ahyong, S.T., Chapple, D.G., Cowan, M., Hossain, M.A., Kennard, M., Macdonald, S., Moore, H., Marsh, J., McCormack, R.B., Michael, D., Mitchell, N., Newell, D., Raadik, T.A. & Tingley, R. (2022) Rapid assessment of the biodiversity impacts of the 2019–2020 Australian megafires to guide urgent management intervention and recovery and lessons for other regions. Diversity and Distributions, 28, 571–591.
    https://doi.org/10.1111/ddi.13428
    Lemckert, F. & Morse, R. (1999) Frogs of the timber production forests of the Dorrigo escarpment in northern New South Wales: an inventory of species present and the conservation of threatened species. In: Campbell, A. (Ed.), Declines and Disappearances of Australian Frogs. Environment Australia, Canberra, pp. 72–80.
    Lemckert, F., Potter, M., Smith, B. & Brues, T. (1997) Recent records of the southern barred frog (Mixophyes balbus) from the south coast of NSW. Herpetofauna, 27, 43–45.
    Littlejohn, M.J. (1969) Amphibians of East Gippsland. Proceedings of the Royal Society of Victoria, 82, 105–12.
    Littlejohn, M. (1987) Calls of Victorian frogs. University of Melbourne, Melbourne.
    Lleonart, J., Salat, J. & Torres, G.J. (2000) Removing allometric effects of body size in morphological analysis. Journal of Theoretical Biology, 205, 85–93.
    https://doi.org/10.1006/jtbi.2000.2043.
    Mahony, M. (1993) The status of frogs in the Watagan mountains area the central coast of New South Wales. In: Lunney, D. & Ayers, D. (Eds.), Herpetology in Australia. Surrey Beatty and Sons Pty Ltd, Sydney, pp. 257–264.
    Mahony, M.J. (2007) Amphibian monitoring in the World Heritage Gondwana Rainforest Reserves of Australia. A report prepared for the Department of Environment and Water, and New South Wales Department of Environment and Climate Change; August 2007.
    Mahony, M.J. (2013) Amphibian Chytrid Fungus Impact Assessment and Management Recommendations. Gondwana Rainforests of Australia World Heritage Area South from Washpool, DECCW-721-2010, pp 1–50.
    Mahony, M., Donnellan, S.C., Richards, S.J. & McDonald, K. (2006) Species boundaries among barred river frogs, Mixophyes (Anura: Myobatrachidae) in north-eastern Australia, with descriptions of two new species. Zootaxa, 1228 (1), 35–60.
    https://doi.org/10.11646/zootaxa.1228.1.3
    Mahony, M.J., Knowles, R., Foster, R. & Donnellan, S.C. (2001) Systematics of the Litoria citropa (Anura: Hylidae) complex in northern New South Wales and southern Queensland, Australia, with the description of a new species. Records of the Australian Museum, 53, 37–48.
    https://doi.org/10.3853/j.0067-1975.53.2001.1322
    Mahony, M.J., Moses, B., Mahony, S., Lemckert, F. & Donnellan, S.C. (2020) A new species of frog in the Litoria ewingii species group (Anura: Pelodryadidae) from south-eastern Australia. Zootaxa, 4858 (2), 201–230.
    https://doi.org/10.11646/zootaxa.4858.2.3
    Mahony, M.J., Penman, T., Bertozzi, T., Lemckert, F., Bilney, R. & Donnellan, S.C. (2021) Taxonomic revision of south-eastern Australian giant burrowing frogs (Anura: Limnodynastidae: Heleioporus Gray). Zootaxa, 5016 (4), 451–489.
    https://doi.org/10.11646/zootaxa.5016.4.1
    Mahony, M., Gould, J., Beranek, C.T., Callen, A., Clulow, J., Clulow, S., Klop-Toker, K., Mahony, S., Wallace, S., Stock, S. & Garnham, J. (2022) A trait-based analysis for predicting impact of wildfires on frogs. Australian Zoologist, 42, 326–351.
    https://doi.org/10.7882/AZ.2022.021
    Mahony, M.J., Hines, H.B., Bertozzi, T., Mahony, S. & Donnellan, S.C. (2022) A sixth species of northern Philoria (Anura: Myobatrachidae) from the uplands of the World Heritage forests of eastern Australia. Zootaxa, 5104 (2), 209–241.
    https://doi.org/10.11646/zootaxa.5104.2.3
    Moritz, C., Hoskin, C.J., MacKenzie, J.B., Phillips, B.L., Tonione, M., Silva, N., Van Der Wal, J., Williams, S.E. & Graham, C.H. (2009) Identification & dynamics of a cryptic suture zone in tropical rainforest. Proceedings of the Royal Society of London, Series B Biological Sciences, 276, 1235–1244.
    https://doi.org/10.1098/rspb.2008.1622
    NSW NEFBS (1994) Fauna of North-east NSW Forests. North-east Forests Biodiversity Study Report No. 3. Unpublished Report, NSW National Parks and Wildlife Service, Hurstville. [unknown pagination]
    Nguyen, L.-T., Schmidt, H.A., von Haeseler, A. & Minh, B.Q. (2015) IQ-TREE: A fast and effective stochastic algorithm for estimating maximum likelihood phylogenies. Molecular Biology Evolution, 32, 268–274.
    https://doi.org/10.1093/molbev/msu300
    Pritchard, J.K., Stephens, M. & Donnelly, P. (2000) Inference of population structure using multilocus genotype data. Genetics, 155, 945–959.
    https://doi.org/10.1534/genetics.116.195164
    Rambaut, A., Drummond, A.J., Xie, D., Baele, G & Suchard, M.A. (2018) Posterior summarisation in Bayesian phylogenetics using Tracer 1.7. Systematic Biology 67: 901–904.
    https://doi.org/10.1093/sysbio/syy032
    Ronquist, F. & Huelsenbeck, J.P. (2003) MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics, 19, 1572–1574.
    https://doi.org/10.1093/bioinformatics/btg180.
    Scheele, B.C., Hollanders, M., Hoffmann, E.P., Newell, D.A., Lindenmayer, D.B., McFadden, M., Gilbert, D.J. & Grogan, L.F. (2021) Conservation translocations for amphibian species threatened by chytrid fungus: A review, conceptual framework, and recommendations. Conservation Science and Practice, 3, e524.
    https://doi.org/10.1111/csp2.524
    Skerratt, L.F., Berger, L., Speare, R., Cashins, S., McDonald, K.R., Phillott, A.D., Hines, H.B. & Kenyon, N. (2007) Spread of chytridiomycosis has caused the rapid global decline and extinction of frogs. EcoHealth, 4, 125.
    https://doi.org/10.1007/s10393-007-0093-5
    Sloan, D.B., Havird, J.C. & Sharbrough, J. (2017) The on-again, off-again relationship between mitochondrial genomes and species boundaries. Molecular Ecology, 26, 2212–2236.
    https://doi.org/10.1111/mec.13959
    Straughan, I.R. (1966) An analysis of the species recognition and species isolation in Queensland frogs. Ph.D Thesis, University of Queensland, St Lucia, Queensland. [unknown pagination]
    Straughan, I.R. (1968) A taxonomic review of the genus Mixophyes (Anura, Leptodactylidae). Proceedings of the Linnean Society of New South Wales, 93, 52–59.
    Swofford, D.L. (2003) PAUP*. Phylogenetic Analysis Using Parsimony (*and Other Methods). Version 4. Sinauer Associates, Sunderland, Massachusetts. [program]
    Thackway, R. & Cresswell, I.D. (1995) An interim biogeographic regionalisation for Australia: a framework for setting priorities in the National Reserves System Cooperative Program. Version 4.0. Australian Nature Conservation Agency, Canberra. [program]
    Thorpe, R.S. (1976) Biometric analysis of geographic variation and racial affinities. Biological Reviews, 51, 407–452.
    https://doi.org/10.1111/j.1469-185X.1976.tb01063.x
    Toews, D.P.L. & Brelsford, A. (2012) The biogeography of mitochondrial and nuclear discordance in animals. Molecular Ecology, 21, 3907–3930.
    https://doi.org/10.1111/j.1365-294X.2012.05664.x
    Trifinopoulos, J., Nguyen, L.-T., von Haeseler, A. & Minh, B.Q. (2016) W-IQ-TREE: a fast online phylogenetic tool for maximum likelihood analysis. Nucleic Acids Research, 44 (W1), W232–W235.
    https://doi.org/10.1093/nar/gkw256
    Tyler, M.J. (1997) The Action Plan for Australian Frogs. Wildlife Australia, Canberra, 77 pp.
    Watson, G.F. & Martin, A.A. (1973) Life history, larval morphology and development of Australian leptodactylid frogs. Transactions of the Royal Society of South Australia, 97, 33–45.
    Wells, K.D. (1977) The courtship of frogs. In: Taylor, D.H. & Guttman, G.I. (Eds.), The reproductive biology of Amphibians. Springer, New York, New York, pp. 233–262.