Integrative taxonomy identifies two new tardigrade species (Eutardigrada: Macrobiotidae) from Greenland

Daniel Stec, Denis T. Tumanov, Reinhardt Møbjerg Kristensen


In this paper we describe Macrobiotus engbergi sp. nov. and Tenuibiotus zandrae sp. nov. from Greenland. Our study has involved both classical taxonomic methods, which include morphological and morphometric analyses conducted with the use of light and scanning electron microscopy, and genetic analysis based on four molecular markers (three nuclear: 18S rRNA, 28S rRNA, ITS-2, and one mitochondrial: COI). Moreover, we re-examined the type series of Tenuibiotus voronkovi (Tumanov, 2007) as well as the original sample where the species was found and we provide new morphological data from light and scanning electron microscopy which enabled us to amend its description. Finally, we also analysed slides with animals and egg of two populations from Nordaustlandet and Edgeøya (archipelago of Svalbard, Norway) designated as T. voronkovi within its recent redescription. The results and comparisons presented in our study question the validity of this designation.



Arctic; egg ornamentation; Macrobiotus hufelandi complex; Tardigrada; Tenuibiotus voronkovi

Full Text:




Bertolani R. & Rebecchi L. 1993. A revision of the Macrobiotus hufelandi group (Tardigrada, Macrobiotidae), with some observations on the taxonomic characters of eutardigrades. Zoologica Scripta 22: 127–152.

Bertolani R., Rebecchi L., Giovannini I. & Cesari M. 2011a. DNA barcoding and integrative taxonomy of Macrobiotus hufelandi C.A.S. Schultze 1834, the first tardigrade species to be described, and some related species. Zootaxa 2997: 19–36.

Bertolani R., Biserov V., Rebecchi L. & Cesari M. 2011b. Taxonomy and biogeography of tardigrades using an integrated approach: new results on species of the Macrobiotus hufelandi group. Invertebrate Zoology 8 (1): 23–36.

Bertolani R., Guidetti R., Marchioro T., Altiero T., Rebecchi L. & Cesari M. 2014. Phylogeny of Eutardigrada: New molecular data and their morphological support lead to the identification of new evolutionary lineages. Molecular Phylogenetics and Evolution 76: 110–126.

Biserov V.I. 1996. Tardigrada of the Novaya Zemlya Archipelago, collected by the Marine Arctic Complex Expedition in 1994. Arthropoda Selecta 5: 151–157.

Biserov V.I. 1999. Tardigrada of Turkmenistan, with description of three new species. Zoologischer Anzeiger 238: 157–167.

Casquet J.T., Thebaud C. & Gillespie R.G. 2012. Chelex without boiling, a rapid and easy technique to obtain stable amplifiable DNA from small amounts of ethanol-stored spiders. Molecular Ecology Resources 12: 136–141.

Cesari M., Bertolani R., Rebecchi L. & Guidetti R. 2009. DNA barcoding in Tardigrada: the first case study on Macrobiotus macrocalix Bertolani & Rebecchi 1993 (Eutardigrada, Macrobiotidae). Molecular Ecology Resources 9 (3): 699–706.

Cesari M., Giovanni I., Bertolani R. & Rebecchi L. 2011. An example of problems associated with DNA barcoding in tardigrades: a novel method for obtaining voucher specimens. Zootaxa 3104: 42–51.

Coughlan K., Michalczyk Ł. & Stec D. 2019. Macrobiotus caelestis sp. nov., a new tardigrade species (Macrobiotidae: hufelandi group) from the Tien Shan Mountains (Kyrgyzstan). Annales Zoologici 69 (3): 499–513.

Coughlan K. & Stec D. 2019. Two new species of the Macrobiotus hufelandi complex (Tardigrada: Eutardigrada: Macrobiotidae) from Australia and India, with notes on their phylogenetic position. European Journal of Taxonomy 573: 1–38.

Dastych H. 1980. Niesporczaki (Tardigrada) Tatrzańskiego Parku Narodowego. Monografie Fauny Polski 9: 1–232.

Degma P. & Guidetti R. 2007. Notes to the current checklist of Tardigrada. Zootaxa 1579: 41–53.

Degma P. & Guidetti R. 2018. Tardigrade taxa. In: Schill, R.O. (ed.) Water Bears: The Biology of Tardigrades: 371–409, Springer, Stuttgart.

Degma P., Bertolani R. & Guidetti R. 2009–2019. Actual checklist of Tardigrada species. Online. Università degli Studi di Modena e Reggio Emilia.

Dudichev A.L. & Biserov V.I. 2000. Tardigrada from Iturup and Paramushir Islands (The Kuril Islands). Zoologicheskii Zhurnal 79: 771–778.

Eibye-Jacobson J. 2001. A New Method for Making SEM Preparations of the Tardigrade Buccopharyngeal Apparatus. Zoologischer Anzeiger 240: 309–319.

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.

Fontoura P., Bartels P.J., Jørgensen A., Kristensen R.M. & Hansen J.G. 2017. A dichotomous key to the genera of the marine heterotardigrades (Tardigrada). Zootaxa 4294 (1): 1–45.

Gąsiorek P., Stec D., Morek W., Zawierucha K., Kaczmarek Ł., Lachowska-Cierlik D. & Michalczyk Ł. 2016. An integrative revision of Mesocrista Pilato, 1987 (Tardigrada: Eutardigrada: Hypsibiidae). Journal of Natural History 50: 2803–2828.

Gąsiorek P., Stec D., Zawierucha Z., Kristensen R.M. & Michalczyk Ł. 2018. Revision of Testechiniscus Kristensen, 1987 (Heterotardigrada: Echiniscidae) refutes the polar-temperate distribution of the genus. Zootaxa 4472 (2): 261–297.

Giribet G., Carranza S., Baguña J., Riutort M. & Ribera C. 1996. First molecular evidence for the existence of a Tardigrada + Arthropoda clade. Molecular Biology and Evolution 13: 76–84.

Grøngaard A., Møbjerg Kristensen N. & Petersen M.K. 1990. Tardigradfaunaen på Disko. In: Kristensen R.M. (ed.) Feltkursus i Arktisk Biologi, Godhavn 1990: 155--180. Zoological Museum, Copenhagen.

Guidetti R. & Bertolani R. 2005. Tardigrade taxonomy: an updated check list of the taxa and a list of characters for their identification. Zootaxa 845: 1–46.

Guidetti R., Gandolfi A., Rossi V. & Bertolani R. 2005. Phylogenetic analysis of Macrobiotidae (Eutardigrada, Parachela): a combined morphological and molecular approach. Zoologica Scripta 34: 235–244.

Guidetti R., Peluffo J.R., Rocha A.M., Cesari M. & Moly de Peluffo M.C. 2013. The morphological and molecular analyses of a new South American urban tardigrade offer new insights on the biological meaning of the Macrobiotus hufelandi group of species (Tardigrada: Macrobiotidae). Journal of Natural History 47 (37–38): 2409–2426.

Guidetti R., Rebecchi L., Bertolani R., Jönsson K.I., Kristensen R.M. & Cesari M. 2016. Morphological and molecular analyses on Richtersius (Eutardigrada) diversity reveal its new systematic position and lead to the establishment of a new genus and a new family within Macrobiotoidea. Zoological Journal of the Linnean Society 178 (4): 834–845.

Guidetti R., Cesari M., Bertolani R., Altiero T. & Rebecchi L. 2019. High diversity in species, reproductive modes and distribution within the Paramacrobiotus richtersi complex (Eutardigrada, Macrobiotidae). Zoological Letters 5: e1.

Guil N. & Giribet G. 2012. A comprehensive molecular phylogeny of tardigrades – adding genes and taxa to a poorly resolved phylum-level phylogeny. Cladistics 28 (1): 21–49.

Guil N., Guidetti R. & Machordom A. 2007. Observations on the “tenuis group” (Eutardigrada, Macrobiotidae) and description of a new Macrobiotus species. Journal of Natural History 41 (41–44): 2741–2755.

Guil N., Jørgensen A. & Kristensen R. 2019. An upgraded comprehensive multilocus phylogeny of the Tardigrada tree of life. Zoologica Scripta 48 (1): 120–137.

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.

Jørgensen A., Kristensen R.M. & Møbjerg N. 2018. Phylogeny and integrative taxonomy of Tardigrada. In: Schill R.O. (ed.) Water Bears: The Biology of Tardigrades: 95–114. Springer, Stuttgart.

Kaczmarek Ł. & Michalczyk Ł. 2017. The Macrobiotus hufelandi (Tardigrada) group revisited. Zootaxa 4363: 101–123.

Kaczmarek Ł., Cytan J., Zawierucha K., Diduszko D. & Michalczyk Ł. 2014. Tardigrades from Peru (South America), with descriptions of three new species of Parachela. Zootaxa 3790: 357–379.

Kaczmarek Ł., Michalczyk Ł. & McInnes S.J. 2016. Annotated zoogeography of non-marine Tardigrada. Part III: North America and Greenland. Zootaxa 4203: 1–249.

Katoh K. & Toh H. 2008. Recent developments in the MAFFT multiple sequence alignment program. Briefings in Bioinformatics 9: 286–298.

Katoh K., Misawa K., Kuma K. & Miyata T. 2002. MAFFT: a novel method for rapid multiple sequence alignment based on fast Fourier transform. Nucleic Acids Research 30: 3059–66.

Kumar S., Stecher G. & Tamura K. 2016. MEGA7: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Molecular Biology and Evolution 33: 1870–1874.

Manicardi G.C. 1989. Two new species of soil moss eutardigrades (Tardigrada) from Canada. Canadian Journal of Zoology 67: 2282–2285.

Maucci W. 1983. Sulla presenza in Italia di Cornechiniscus holmeni (Petersen, 1951), e descrizione di Macrobiotus hyperonyx sp. nov. Bollettino del Museo civico di Storia naturale di Verona 9: 175–179.

Maucci W. 1988. Tardigradi della Mongolia Esterna, con descrizione di Macrobiotus mongolicus sp. nov. Bollettino del Museo civico di Storia naturale di Verona 14: 339–349.

Maucci W. 1996. Tardigrada of the Arctic tundra with descriptions of two new species. Zoological Journal of the Linnean Society 116: 185–204.

Michalczyk Ł. & Kaczmarek Ł. 2003. A description of the new tardigrade Macrobiotus reinhardti (Eutardigrada, Macrobiotidae, harmsworthi group) with some remarks on the oral cavity armature within the genus Macrobiotus Schultze. Zootaxa 331: 1–24.

Michalczyk Ł. & Kaczmarek Ł. 2013. The Tardigrada Register: a comprehensive online data repository for tardigrade taxonomy. Journal of Limnology 72: 175–181.

Michalczyk Ł., Wełnicz W., Frohme M. & Kaczmarek Ł. 2012. Redescriptions of three Milnesium Doyère, 1840 taxa (Tardigrada: Eutardigrada: Milnesiidae), including the nominal species for the genus. Zootaxa 3154: 1–20.

Mironov S.V., Dabert J., Dabert M. 2012. A new feather mite species of the genus Proctophyllodes Robin, 1877 (Astigmata: Proctophyllodidae) from the Long-tailed Tit Aegithalos caudatus (Passeriformes: Aegithalidae): morphological description with DNA barcode data. Zootaxa 3253: 54–61.

Møbjerg N., Jørgensen A., Kristensen R.M. & Neves R.C. 2018. Morphology and functional anatomy. In: Schill R.O. (ed.) Water Bears: The Biology of Tardigrades 57–94. Springer, Stuttgart.

Morek W., Stec D., Gąsiorek P., Schill R.O., Kaczmarek Ł. & Michalczyk Ł. 2016. An experimental test of eutardigrade preparation methods for light microscopy. Zoological Journal of the Linnean Society 178: 785–793.

Nelson D.R., Guidetti R. & Rebecchi L. 2015. Phylum Tardgrada. In: Thorp and Covich’s Freshwater Invertebrates: 347–380, Academic Press, New York.

Nowak B. & Stec D. 2018. An integrative description of Macrobiotus hannae sp. nov. (Tardigrada: Eutardigrada: Macrobiotidae: hufelandi group) from Poland. Turkish Journal of Zoology 42: 269–286.

Pilato G. 1981. Analisi di nuovi caratteri nello studio degli Eutardigradi. Animalia 8: 51–57.

Roszkowska M., Ostrowska M., Stec D., Janko K. & Kaczmarek Ł. 2017. Macrobiotus polypiformis sp. nov., a new tardigrade (Macrobiotidae; hufelandi group) from the Ecuadorian Pacific coast, with remarks on the claw abnormalities in eutardigrades. European Journal of Taxonomy 327: 1–19.

Schill R.O., Forster F., Dandekar T. & Wolf N. 2010. Using compensatory base change analysis of internal transcribed spacer 2 secondary structures to identify three new species in Paramacrobiotus (Tardigrada). Organisms Diversity & Evolution 10 (4): 287–296.

Stec D. 2019. Mesobiotus datanlanicus sp. nov., a new tardigrade species (Macrobiotidae: Mesobiotus harmsworthi group) from Lâm Đồng Province in Vietnam. Zootaxa 4679 (1): 164–180.

Stec D., Smolak R., Kaczmarek Ł. & Michalczyk Ł. 2015. An integrative description of Macrobiotus paulinae sp. nov. (Tardigrada: Eutardigrada: Macrobiotidae: hufelandi group) from Kenya. Zootaxa 4052: 501–526.

Stec D., Gąsiorek P., Morek W., Kosztyła P., Zawierucha K., Michno K., Kaczmarek Ł., Prokop Z.M. & Michalczyk Ł. 2016. Estimating optimal sample size for tardigrade morphometry. Zoological Journal of the Linnean Society 178: 776–784.

Stec D., Zawierucha K. & Michalczyk Ł. 2017a. An integrative description of Ramazzottius subanomalus (Biserov, 1985) (Tardigrada) from Poland. Zootaxa 4300 (3): 403–420.

Stec D., Morek W., Gąsiorek P., Blagden B. & Michalczyk Ł. 2017b. Description of Macrobiotus scoticus sp. nov. (Tardigrada: Macrobiotidae: hufelandi group) from Scotland by means of integrative taxonomy. Annales Zoologici 67: 181–197.

Stec D., Morek W., Gąsiorek P. & Michalczyk Ł. 2018a. Unmasking hidden species diversity within the Ramazzottius oberhaeuseri complex, with an integrative redescription of the nominal species for the family Ramazzottiidae (Tardigrada: Eutardigrada: Parachela). Systematics and Biodiversity 16 (4): 357–376.

Stec D., Krzywański Ł. & Michalczyk Ł. 2018b. Integrative description of Macrobiotus canaricus sp. nov. with notes on M. recens (Eutardigrada: Macrobiotidae). European Journal of Taxonomy 452: 1–36.

Stec D., Kristensen R.M. & Michalczyk Ł. 2018c. Integrative taxonomy identifies Macrobiotus papei, a new tardigrade species of the Macrobiotus hufelandi complex (Eutardigrada: Macrobiotidae) from the Udzungwa Mountains National Park (Tanzania). Zootaxa 4446: 273–291.

Stec D., Arakawa K. & Michalczyk Ł. 2018d. An integrative description of Macrobiotus shonaicus sp. nov. (Tardigrada: Macrobiotidae) from Japan with notes on its phylogenetic position within the hufelandi group. PLoS ONE 13: e0192210.

Stec D., Krzywański Ł., Zawierucha K. & Michalczyk Ł. 2020. Untangling systematics of the Paramacrobiotus areolatus species complex by an integrative redescription of the nominal species for the group, with multilocus phylogeny and species delineation within the genus Paramacrobiotus. Zoological Journal of the Linnean Society.

Sutcliffe A.J. & Blake W. Jr. 2000. Biological activity on a decaying caribou antler at Cape Herschel, Ellesmere Island, Nunavut, High Arctic Canada. Polar Record 36 (198): 233–246.

Tumanov D.V. 2007. Three new species of Macrobiotus (Eutardigrada, Macrobiotidae, tenuis-group) from Tien Shan (Kirghizia) and Spitsbergen. Journal of Limnology 66: 40–48.

Wełnicz W., Grohme M.A., Kaczmarek Ł., Schill R.O. & Frohme M. 2011. ITS-2 and 18S rRNA data from Macrobiotus polonicus and Milnesium tardigradum (Eutardigrada, Tardigrada). Journal of Zoological Systematics and Evolutionary Research 49 (S1): 34–39.

Zawierucha K., Coulson J., Michalczyk Ł. & Kaczmarek Ł. 2013. Current knowledge on the Tardigrada of Svalbard with the first records of water bears from Nordaustlandet (High Arctic). Polar Research 32: e20886.

Zawierucha K., Kolicka M. & Kaczmarek Ł. 2016a. Re-description of the Arctic tardigrade Tenuibiotus voronkovi (Tumanov, 2007) (Eutardigrada; Macrobiotidea), with the first molecular data for the genus. Zootaxa 4196 (4): 498–510.

Zawierucha K., Zmudczyńska-Skarbek K., Kaczmarek Ł. & Wojczulanis-Jakubas K. 2016b. The influence of a seabird colony on abundance and species composition of water bears (Tardigrada) in Hornsund (Spitsbergen, Arctic). Polar Biology 39: 713–723.

Zeller C. 2010. Untersuchung der Phylogenie von Tardigraden anhand der Genabschnitte 18S rDNA und Cytochrom c Oxidase Untereinheit 1 (COX I). MSc Thesis, Technische Hochschule Wildau, Germany.


  • There are currently no refbacks.

License URL: