An integrative description of Macrobiotus hannae sp. nov. (Tardigrada: Eutardigrada: Macrobiotidae: hufelandi group) from Poland

A new species of the Macrobiotus hufelandi group is described from Poland. An integrative taxonomic approach was applied by combining morphological and morphometric analysis imaging under phase contrast and scanning electron microscopy with molecular analysis (four molecular markers) to unambiguously support the establishment of the new species. The specimens of Macrobiotus hannae sp. nov. are similar to three taxa of the Macrobiotus hufelandi complex: Macrobiotus hufelandi C.A.S. Schultze, 1834; M. punctillus Pilato, Binda & Azzaro, 1990; and M. joannae Pilato & Binda, 1983. Nevertheless, the new species differs from the first two species by the absence of cuticular bars under claws on legs I?III, better developed oral cavity armature (large, with all three bands of teeth always well visible), the presence of a subterminal constriction in the second macroplacoid, some fine peculiarities in egg ornamentation morphology, and differences in a number of morphometric characters. The new species is most similar to M. joannae but it can be distinguished from it by the presence of a much finer granulation on the dorsal and lateral cuticle of the body that, in contrast to M. joannae, is not visible in light microscopy. Our study suggests that European records of M. joannae are most likely invalid and they represent M. hannae or an unknown similar species.

___

  • Bertolani R (1971). Rapporto-sessi e dimorfismo sessuale in Macrobiotus (Tardigrada) Rend. Accademia Nazionalle Lincei Ser 8 50: 377-382 (in Italian).
  • Bertolani R, Biserov V, Rebecchi L, Cesari M (2011a). Taxonomy and biogeography of tardigrades using an integrated approach: new results on species of the Macrobiotus hufelandi group. Invert Zool 8: 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. Mol Phylogenet Evol 76: 110-126.
  • Bertolani R, Pilato G, Sabatini MA (1983). Macrobiotus joannae , primo Macrobiotidae ermafrodito (Eutardigrada). Animalia 10: 327-333 (in Italian).
  • Bertolani R, Rebecchi L (1993). A revision of the Macrobiotus hufelandi group (Tardigrada, Macrobiotidae), with some observations on the taxonomic characters of eutardigrades. Zool Scr 22: 127-152.
  • Bertolani R, Rebecchi L, Giovannini I, Cesari M (2011b). 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.
  • Binda MG, Pilato G (1984). Macrobiotus sapiens , nuova specie di Eutardigrado di Sicilia. Animalia 11: 85-90 (in Italian).
  • Biserov VI (1990). On the revision of the genus Macrobiotus . The subgenus Macrobiotus s. st. is a new systematic status of the hufelandi group (Tardigrada, Macrobiotidae). Communication, 2. Zool Zhurnal 69: 38-50.
  • Casquet J, Thebaud C, Gillespie RG (2012). Chelex without boiling, a rapid and easy technique to obtain stable amplifiable DNA from small amounts of ethanol-stored spiders. Mol Ecol Resour 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). Mol Ecol Resour 9: 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.
  • Dastych H (1980). Niesporczaki (Tardigrada) Tatrzańskiego Parku Narodowego. Monografie Fauny Polski 9: 1-232 (in Polish).
  • Dastych H (1988). The Tardigrada of Poland. Monografie Fauny Polski 16: 1-255.
  • Dastych H (1997). Niesporczaki – Tardigrada. In: Razowski J, editor. Checklist of Animals of Poland, Vol. 4. Kraków, Poland: Institute of Systematics and Evolution of Animals, pp. 141-144.
  • Doyère LMF (1840). Memoire sur les Tardigrades. I. Ann Sci Nat Paris Series 2 14: 269-362 (in French).
  • 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. Mol Mar Biol Biotechnol 3: 294-299.
  • 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). J Nat Hist 50: 2803-2828.
  • Giribet G, Carranza S, Baguna J, Riutort M, Ribera C (1996). First molecular evidence for the existence of a Tardigrada + Arthropoda clade. Mol Biol Evol 13: 76-84.
  • Guidetti R, Gandolfi A, Rossi V, Bertolani R (2005). Phylogenetic analysis of Macrobiotidae (Eutardigrada, Parachela): a combined morphological and molecular approach. Zool Scr 34: 235-244.
  • Guidetti R, Peluffo JR, Rocha AM, Cesari M, Moly de Peluffo MC (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). J Nat Hist 47: 2409-2426.
  • Guil N, Giribet G (2012). A comprehensive molecular phylogeny of tardigrades – adding genes and taxa to a poorly resolved phylum-level phylogeny. Cladistics 28: 21-49.
  • Hall TA (1999). BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/ NT. Nucleic Acids Symposium Series 41: 95-98.
  • Hebert PDN, Cywinska A, Ball SL, deWaard JR (2003). Biological identifications through DNA barcodes. Proc R Soc London Ser B Biol Sc 270: 313-322.
  • Hęciak S (1976). Niesporczaki (Tardigrada) Gór Świętokrzyskich. Bad Fizjogr Pol Zach 29: 111-128 (in Polish).
  • Jakubski A (1915). Opis fauny wrotków (Rotatoria) powiatu sokalskiego z uwzględnieniem gromad brzuchorzęsków (Gartroprioga) i niesporczaków (Tardigrada). Wiadom Mus Dzieduszyckich Lwów 1: 1-166 (in Polish).
  • 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, Kosicki J, Roszkowska M (2018). Tardigrada of Bory Tucholskie National Park, Zaborski Landscape Park and their surroundings (Pomerania Province, Poland). Turk J Zool 42: 6-17.
  • Kaczmarek Ł, Michalczyk Ł (2004). Macrobiotus macrocalix Bertolani & Rebecchi, 1993 – a species of water bear (Tardigrada) new to the fauna of Poland. Bad Fizjogr Pol Zach 50: 39-43.
  • Kaczmarek Ł, Michalczyk Ł (2017). The Macrobiotus hufelandi (Tardigrada) group revisited. Zootaxa 4363: 101-123.
  • 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 Res 30: 3059-3066.
  • Katoh K, Toh H (2008). Recent developments in the MAFFT multiple sequence alignment program. Brief Bioinform 9: 286-298.
  • Kumar S, Stecher G, Tamura K (2016). MEGA7: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Mol Biol Evol 33: 1870-1874.
  • Marley NJ, McInnes SJ, Sands CJ (2011). Phylum Tardigrada: a re- evaluation of the Parachela. Zootaxa 2819: 51-64.
  • Michalczyk Ł, Kaczmarek Ł (2003a). Water bears (Tardigrada) of the Bieszczady Mountains (Poland, West Carpathians). Zootaxa 242: 1-24.
  • Michalczyk Ł, Kaczmarek Ł (2003b). 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. J Limnol 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.
  • Minkiewicz S (1914). Przegląd fauny jezior tatrzańskich. Akademia Umiej ętności (Krak ów) Komisja Fizjograficzna 48: 114-137 (in Polish).
  • Mironov SV, 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.
  • Morek W, Stec D, Gąsiorek P, Schill R O, Kaczmarek Ł, Michalczyk Ł (2016). An experimental test of eutardigrade preparation methods for light microscopy. Zool J Linn Soc-Lond 178: 785- 793.
  • Nelson DR, Guidetti R, Rebecchi L (2015). Phylum Tardigrada. In: Thorp J, Rogers DC, editors. Ecology and General Biology: Vol. 1: Thorp and Covich’s Freshwater Invertebrates. 4th Revised Edition. New York, NY, USA: Academic Press, pp. 347-380.
  • Nowak B, Stec D (2017). The first record of Macrobiotus vladimiri Bertolani, Biserov, Rebecchi and Cesari, 2011 (Tardigrada: Eutardigrada: Macrobiotidae: hufelandi group) from Poland. Turk J Zool 41: 558-567.
  • Pigoń A, Węglarska B (1953). The respiration of Tardigrada: a study in animal anabiosis. Bull Acad Pol Sci 1: 69-72.
  • Pilato G (1981). Analisi di nuovi caratteri nello studio degli Eutardigrada. Animalia 8: 51-57 (in Italian).
  • Pilato G, Binda MG (1983). Descrizione du una nuova specie di Eutardigrado d’Australia Macrobiotus joannae n. sp. Animalia 10: 262-272 (in Italian).
  • Pilato G, Binda MG, Azzaro M (1990). Tardigradi di Terra del Fuoco e Magallanes. III. Macrobiotus punctillus , nuova specie di Macrobiotidae del gruppo hufelandi . Animalia 17: 123-129 (in Italian).
  • Pilato G, Dastych H (1974). Diphascon montigenum sp. nov., a new species of Tardigrada from Poland. Bull Acad Pol Sci 22: 325- 327.
  • Pilato G, Kaczmarek Ł, Michalczyk Ł, Lisi O (2003). Macrobiotus polonicus , a new species of Tardigrada from Poland (Eutardigrada, Macrobiotidae, ‘ hufelandi group’). Zootaxa 258: 1-8.
  • Pilato G, Kiosya Y, Lisi O, Sabella G (2012). New records of Eutardigrada from Belarus with the description of three new species. Zootaxa 3179: 39-60.
  • Richters F (1926). Tardigrada. In: Kükenthal W, Krumbach T, editors. Handbuch der Zoologie Vol. 3. Berlin, Germany: Walter de Gruyter & Co., pp. 58-61 (in German).
  • 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. Eur J Taxon 327: 1-19.
  • Schill RO, 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). Org Divers Evol 10: 287-296.
  • Schultze CAS (1834). Macrobiotus Hufelandii animal e crustaceorum classe novum, reviviscendi post diuturnam asphixiam et aridiatem potens, etc. 8, 1 tab. C. Berlin, Germany: Curths (in Latin).
  • Schuster RO, Nelson DR, Grigarick AA, Christenberry D (1980). Systematic criteria of the Eutardigrada. T Am Microsc Soc 99: 284-303.
  • Srivathsan A, Meier R (2012). On the inappropriate use of Kimura-2- parameter (K2P) divergences in the DNA-barcoding literature. Cladistics 28: 190-194.
  • Stec D, Gąsiorek P, Morek W, Kosztyła P, Zawierucha K, Michno K, Kaczmarek Ł, Prokop ZM, Michalczyk Ł (2016a). Estimating optimal sample size for tardigrade morphometry. Zool J Linn Soc-Lond 178: 776-784.
  • Stec D, Morek W, Gąsiorek P, Blagden B, Michalczyk Ł (2017a). 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, Kaczmarek Ł, Michalczyk Ł (2016b). Determinants and taxonomic consequences of extreme egg shell variability in Ramazzottius subanomalus (Biserov, 1985) (Tardigrada). Zootaxa 4208: 176-188.
  • Stec, D Morek W, Gąsiorek P, Michalczyk Ł (2018). Unmasking hidden species diversity within the Ramazzottius oberhaeuseri complex, with an integrative redescription of the nominal species for the family Ramazzottiidae (Tardigrada: Eutardigrada: Parachela). Syst Biodivers (in press).
  • 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, Zawierucha K, Michalczyk Ł (2017b). An integrative description of Ramazzottius subanomalus (Biserov, 1985) (Tardigrada) from Poland. Zootaxa 4300: 403-420.
  • Thulin G (1928). Über die Phylogenie und das System der Tardigraden. Hereditas 11: 207-266 (in German).
  • Węglarska B (1959). Tardigraden Polens II. Vest Csl Spol Zool 23: 354-357 (in German).
  • Węglarska B (1973). Tardigrada in High Tatra localities bare of snow with a description of Itaquascon pawlowskii sp. nov. Vest Cs Spol Zool 37: 150-154.
  • Węglarska B, Korecka T (1983). Tardigrada from Dobczyce area (Poland). Zes Nauk Univ Jagiell 29: 83-92.
  • Wełnicz W, Grohme MA, Kaczmarek Ł, Schill RO, Frohme M (2011). ITS-2 and 18S rRNA data from Macrobiotus polonicus and Milnesium tardigradum (Eutardigrada, Tardigrada). J Zool Syst Evol Res 49: 34-39.
  • Zawierucha K, Kolicka M, Takeuchi N, Kaczmarek Ł (2015). What animals can live in cryoconite holes? A faunal review. J Zool 295: 159-169.
  • Zeller C (2010). Untersuchung der Phylogenie von Tardigraden anhand der Genabschnitte 18S rDNA und Cytochrom c Oxidase Untereinheit 1 (COX I). MSc, Technische Hochschule Wildau, Wildau, Germany (in German).