Notes on the earthworm species, Metaphire anomala (Michaelsen, 1907) (Clitellata, Megascolecidae) in Southern Vietnam, with descriptions of two new species

Keywords: Metaphire, taxonomy, biodiversity, COI genetic divergence, Vietnam


Integrative taxonomy was employed to exploit the differences between the known Metaphire anomala (Michaelsen, 1907) and other specimens collected in Vietnam. The results brought to light two new species, namely Metaphire iranomala sp. nov. and Metaphire decemtheca sp. nov. The former is easily recognised by having male pores on xix and four pairs of spermathecal pores on 5/6/7/8/9 while the latter is distinguished by having five pairs of spermathecal pores on 4/5/6/7/8/9. The K2P distances of the fragment of the cytochrome c oxidase subunit I (COI) gene are 13.1% between M. iranomala sp. nov. and M. anomala (Michaelsen, 1907) and 18% between M. decemtheca sp. nov. and Metaphire grandiverticulata Nguyen & Lam, 2017. The intraspecific divergences are 1.5%–10.6% for M. iranomala sp. nov. and 2.1%–11.4% for M. decemtheca sp. nov.


Aspe N.M. & James S.W. 2018. Molecular phylogeny and biogeographic distribution of pheretimoid earthworms (Clitellata: Megascolecidae) of the Philippine archipelago. European Journal of Soil Biology 85: 89–97.

Bantaowong U., Chanabun R., Tongkerd P., Sutcharit C., James S.W. & Panha S. 2011. A new species of the terrestrial earthworm of the genus Metaphire Sims and Easton, 1972 from Thailand with redescription of some species. Tropical Natural History 11 (1): 55–69.

Blakemore R.J. 2016a. Darwin’s earthworms (Annelida, Oligochaeta, Megadrilacea) with review of cosmopolitan Metaphire peguana-species gr oup from Philippines. Opuscula Zoologica 47 (1): 9–30.

Blakemore R.J. 2016b. Eco-taxonomic profile and mtDNA barcode of Metaphire megascolidioides (Goto & Hatai, 1899) – a megadrile earthworm from Japan, plus miscellaneous taxonomic mending. VermEcology Occasional Papers 1: 1–16.

Chang C.-H., Lin S.-M. & Chen J.-H. 2008. Molecular systematics and phylogeography of the gigantic earthworms of the Metaphire formosae species group (Clitellata, Megascolecidae). Molecular Phylogenetics and Evolution 49: 958–968.

Chen Y. 1938. Oligochaeta from Hainan, Kwangtung. Contributions from the Biological Laboratory of the Science Society of China. Zoological Series 12 (10): 375–427.

Cognetti M.L. 1908. Contributo alla conscenza della drilofauna papuasica. Bollettino dei musei di zoologia ed anatomia comparata della R. Università di Torino 23 (584): 1–4.

Edgar R.C. 2004. MUSCLE: a multiple sequence alignment method with reduced time and space complexity. BMC Bioinformatics 5: 113.

Feldman A.T. & Wolfe D. 2014. Tissue processing and hematoxylin and eosin staining. In: Day C. (ed.) Histopathology. Methods in Molecular Biology (Methods and Protocols) 1180: 31–43. Humana Press, New York.

Folmer O., Black M., Hoeh W., Lutz R. & Vrijenhoek R. 1994. DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular Marine Biology and Biotechnology 3: 294–299.

Gates G.E. 1925. Some notes on Pheretima anomola, Mich. and a related species new to India and Burma. Annals and Magazine of Natural History 15 (9): 538–550.

Gates G.E. 1972. Burmese earthworms: an introduction to the systematics and biology of Megadrile oligochaetes with special reference to Southeast Asia. Transactions of the American Philosophical Society 62: 1–326.

Goto S. & Hatai S. 1899. New or imperfectly known species of earthworm. Annotationes Zoologicae Japonenses 3 (2): 13–24.

Hall T.A. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for windows 95/98/NT. Nucleic Acids Symposium Series 41: 95–98.

James S.W., Porco D., Decaens T., Richard B., Rougerie R. & Ersaeus C. 2010. DNA barcoding reveals cryptic diversity in Lumbricus terrestris L., 1758 (Clitellata): resurrection of L. herculeus (Savigny, 1826). PLoS One 5: e15629.

Jeratthitikul E., Bantaowong U. & Panha S. 2017. DNA barcoding of the Thai species of terrestrial earthworms in the genera Amynthas and Metaphire (Haplotaxida: Megascolecidae). European Journal of Soil Biology 81: 39–47.

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.

King R.A., Tibble A.L. & Symondson W.O.C. 2008. Opening a can of worms: unprecedented sympatric cryptic diversity within British lumbricid earthworms. Molecular Ecology 17: 4684–4698.

Klarica J., Kloss-Brandsttätter A., Traugott M. & Juen A. 2012. Comparing four mitochondrial genes in earthworms – implications for identification, phylogenetics, and discovery of cryptic species. Soil Biology and Biochemistry 45: 23–30.

Kumar S., Stecher G. & Tamura K. 2016. MEGA 7: Molecular Evolution Genetics Analysis version 7.0 for bigger datasets. Molecular Biology and Evolution 33: 1870–1874.

Michaelsen W. 1907. Neue Oligochäten von Vorder-Indien, Ceylon, Burma und den Andaman-Inseln. Mitteilungen aus dem naturhistorischen Museum in Hamburg 24: 143–193.

Minamiya Y., Yokoyama J. & Fukuda T. 2009. A phylogeographic study of the Japanese earthworm, Metaphire sieboldi (Horst, 1883) (Oligochaeta: Megascolecidae): inferences from mitochondrial DNA sequences. European Journal of Soil Biology 45 (5/6): 423–430.

Minh B.Q., Schmidt H.A., Chernomor O., Schrempf D., Woodhams M.D., von Haeseler A. & Lanfear R. 2020. IQ-TREE 2: new models and efficient methods for phylogenetic inference in the genomic era. Molecular and Biological Evolution 37: 1530–1534.

Nguyen A.D. & Nguyen T.T. 2015. Notes on Metaphire multitheca (Chen, 1938) (Oligochaeta, Megascolecidae) recorded from Vietnam, with descriptions of two new species. ZooKeys 506: 127–136.

Nguyen Q.N., Nguyen V.T., Duong C.T., Le V.N. & Nguyen T.T. 2019. Species diversity of earthworms in Dong Nai Province, Vietnam. Tap Chi Sinh Hoc 41 (2): 117–129.

Nguyen T.T. 2014. Checklist and some remarks on faunistic characteristics of earthworms in the Mekong Delta, Vietnam. Journal of Science, Cantho University, Section A: Science, Technology and Environment 32: 106–119.

Nguyen T.T & Lam H.D. 2017. Three new earthworm species of the genus Metaphire Sims & Easton, 1972 (Oligochaeta, Megascolecidae) from Dong Nai Province, Vietnam. Tap Chi Sinh Hoc 39 (4): 406–415.

Nguyen T.T., Nguyen A.D., Tran T.T.B. & Blakemore R.J. 2016. A comprehensive checklist of earthworm species and subspecies from Vietnam (Annelida: Clitellata: Oligochaeta: Almidae, Eudrilidae, Glossoscolecidae, Lumbricidae, Megascolecidae, Moniligastridae, Ocnerodrilidae, Octochaetidae). Zootaxa 4140: 1–92.

Nguyen T.T., Trinh K.B.T., Nguyen H.L.T. & Nguyen A.D. 2017a. Earthworms (Annelida: Oligochaeta) from islands of Kien Hai District, Kien Giang Province, Vietnam, with descriptions of two new species and one subspecies. Journal of Natural History 51 (15–16): 883–915.

Nguyen T.T., Nguyen P.H., Truong T.A. & Nguyen Q.N. 2017b. Diversity and distribution of earthworms in Ba Ria, Vung Tau Province. Journal of Science, Can Tho University 53 (A): 96–107.

Nguyen T.T., Nguyen Q.N & Nguyen A.D. 2018. The first record of the earthworm genus Pheretima Kinberg, 1867 sensu stricto in Vietnam, with description of a new species (Annelida: Clitellata: Megascolecidae). Zootaxa 4496 (1): 251–258.

Nguyen T.T., Lam D.H., Trinh B.T.K. & Nguyen A.D. 2020. The megascolecid earthworms (Annelida, Oligochaeta, Megascolecidae) in the Phu Quoc island, Vietnam, with descriptions of three new species. ZooKeys 932: 1–25.

Novo M., Almodovar A., Fernandez R., Trigo D. & Díaz-Cosín D.J. 2010. Cryptic speciation of hormogastrid earthworms revealed by mitochondrial and nuclear data. Molecular Phylogenetics and Evolution 56: 507–512.

Prasankok P., Bantaowong U., James S.W. & Panha S. 2013. Low heterogeneity in populations of the terrestrial earthworm, Metaphire peguana (Rosa, 1890), in Thailand, as revealed by analysis of mitochondrial DNA CO1 sequences and nuclear allozymes. Biochemical Systematics and Ecology 51: 8–15.

Römbke J., Aira M., Backeljau T., Breugelmans K., Domínguez J., Funke E., Graf N., Hajibabaei M., Perez-Losada M., Porto P.G., Schmelz R.M., Vierna J., Vizcaíno A. & Pfenninger M. 2016. DNA barcoding of earthworms (Eisenia fetida/andrei complex) from 28 ecotoxicological test laboratories. Applied Soil Ecology 104: 3–11.

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 selection across a large model space. Systematic Biology 61: 539–542.

Rougerie R., Decaens T., Deharveng L., Porcol D., James S.W., Chang C.-H., Richard B., Potapov M., Suhardjono Y. & Hebert P.D.N. 2009. DNA barcodes for soil animal taxonomy. Pesquisa Agropecuária Brasileira 44: 789–801.

Sims R.W. & Easton E.G. 1972. A numerical revision of the earthworm genus Pheretima auct. (Megascolecidae: Oligochaeta) with the recognition of new genera and an appendix on the earthworms collected by the Royal Society North Borneo Expedition. Biological Journal of the Linnean Society 4: 169–268.

Stephenson J. 1929. The Oligochaeta of the lndawgyi lake (upper Burma). Records of the Indian Museum 31: 225–239.

Thai T.B., Huynh T.K.H. & Nguyen D.A. 2004. Remarks of earthworms on the islands in southern of Vietnam. Proceedings of the national Workshop on the basic Issues in Life Science. Hanoi Science and Technics Publishing House: 757–760.

Yuan Z., Dong Y., Jiang J., Jiang J., Zhao Q. & Qiu J. 2019. Three new species of earthworms (Oligochaeta: Megascolecidae) from Yunnan Province, China. Zootaxa 4664 (3): 390–400.

Zhang Z., Schwartz S., Wagner L. & Miller W. 2000. A greedy algorithm for aligning DNA sequences. Journal of Computational Biology 7 (1/2): 203–214.

How to Cite
Nguyen, T. T., Lam, D. H., & Nguyen, A. D. (2021). Notes on the earthworm species, Metaphire anomala (Michaelsen, 1907) (Clitellata, Megascolecidae) in Southern Vietnam, with descriptions of two new species. European Journal of Taxonomy, 746(1), 94-111.
Research article