Abstract
Hungarosoma bokori Verhoeff, 1928 is a millipede species which was originally classified solely on the basis of a female specimen. Subsequently, a long history of field searching for and surmising about the systematic position of this small, enigmatic species followed. In April 2013, 85 years after its first description, a series of nine specimens were sampled in the type locality, the Abaliget Cave, in southern Hungary. An adult male was collected for the first time, along with females and juveniles. Descriptions of the gonopods and the female vulvae, both important for considerations of the systematic position of the species, are presented for the first time. Revision and re-designation of the type material was made.
The cryptic life of the species is connected with its activity in winter, and its known fragmented distribution corresponds with its presence in undisturbed microhabitats having a specific microclimate, often in the soil at cave entrances.
Molecular methods showed a positive detection of the intracellular prokaryotic parasite Wolbachia in H. bokori, reflecting its highly probable parthenogenetic character in the main part of its known area of occurrence. This is the first demonstration of Wolbachia in a millipede.
The legitimacy of the family Hungarosomatidae Ceuca, 1974, as a separate taxon was analysed using morphological and molecular approaches. Results of both methods confirmed the existence of a distinct phyletic line. DNA barcoding has shown its closest position to Attemsiidae Verhoeff, 1899, or Neoatractosomatidae Verhoeff, 1901. Based on records from Austria, the Czech Republic, Hungary and Slovakia, the residual circum-pannonic distribution that the whole genus (family) probably represents is proposed.
References
Angyal, D. & Korsós, Z. (2013) Millipedes (Diplopoda) of twelve caves in Western Mecsek, Southwest Hungary. Opuscula Zoologica, Budapest, 44, 99–106.
Blower, G.J. (1985) Millipedes. Keys and notes for the identification of the species. Linnean Society of London and The Estuarine and Brackish-Water Sciences Association by E.J.Brill/Dr.W. Backhuys, London, 242 pp.
Brolemann, H.W. (1935) Myriapodes diplopodes (Chilognathes I). Faune de France, 29. P. Lechevalier, Paris, 369 pp.
Ceuca, D. (2008) Des diplopodes des zones de Cluj. Oltenia - Studii şi comunicări Ştiințele Naturii, 24, 48–55.
Ceuca, T. (1967) Quelques autres Diplopodes nouveaux de la faune de la Roumanie. Studia Universitatis Babeş-Bolyai. Series Biologia, 1, 107–117.
Ceuca, T. (1974) Noi contribuţii la cunoaşterea diplopodelor din fauna României. Studia Universitatis Babeş-Bolyai. Series Biologia, 1, 91–97.
Ceuca, T. (1992) Quelques aspects sur la faunistique, l’écologie et la zoogeographie des Diplopodes de la Région Balkanique. Berichte des Naturwissenschaftlich-Medizinischen Vereins in Innsbruck, Supplementum, 10, 411–429.
Cook, O.F. (1895) Introductory note on the families of Diplopoda. In: Cook, O.F. & Collins, G.N. (Eds.), The Craspedosomatidae of North America. Annals of the New York Academy of Sciences, 9, 1–7.
http://dx.doi.org/10.1111/j.1749-6632.1896.tb55430.xCsuzdi, C., Pop, V.V. & Pop, A.A. (2011) The earthworm fauna of the Carpathian Basin with new records and description of three new species (Oligochaeta: Lumbricidae). Zoologischer Anzeiger, 250, 2–18.
http://dx.doi.org/10.1016/j.jcz.2010.10.001Ćurčić, B.P.M., Makarov, S.E., Tomić, V.T. & Mitić, B.M. (2007) Global review of some Anthroleucosomatidae (Diplopoda: Chordeumatida) with description of a new millipede genus from Balkan Peninsula. Canadian Entomologist, 139, 478–488.
http://dx.doi.org/10.4039/n06-062Dudich, E. (1932) Biologie der Aggteleker Tropfsteinhöhle “Baradla” in Ungarn. Speläologische Monographien, 13, Wien, 246 pp.
Latzel, R. (1884) Die Myriapoden der Österreichisch-Ungarischen Monarchie. Band 2. Wien, 414 pp.
Enghoff, H. (2013) Diplopoda. Fauna Europaea version 2.6.2 – last update 29 August 2013. Available from: http://www.faunaeur.org (accessed 19 August 2015)
Enghoff, H. & Golovatch, S.I. (2015) Order Chordeumatida Pocock, 1894. In: Minelli, A. (Ed.) The Myriapoda. Treatise on Zoology –Anatomy, Taxonomy, Biology. Volume 2. Brill, Leiden – Boston, pp. 408–420.
Felsenstein, J. (1985) Confidence limits on phylogenies: An approach using the bootstrap. Evolution, 39, 783–791.
http://dx.doi.org/10.2307/2408678Folmer, 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.
Gebhardt, A. (1933) Az Abaligeti és a Mánfai barlang állatvilágának összehasonlítása (Vergleichung der Tierwelt der Abaligeter- und Mánfaer Höhlen). Állatani Közlemények, 30, 36–44.
Gebhardt, A. (1934) Az abaligeti barlang élővilága (The fauna of the Abaliget Cave). Mathemathikai és Természettudományi Közlemények, 37, 1–264 + map.
Gebhardt, A. (1963) A Mecsek-hegység barlangjainak biológiai vizsgálata. Janus Pannonius Múzeum Évkönyve, 8, 5–32.
Gebhardt, A. (1967) A Mecsek hegység állatvilágának térbeli elterjedése élőhelyek szerint. Janus Pannonius Múzeum Évkönyve, 12, 7–14.
Golovatch, S.I. & Kime, R.D. (2009) Millipede (Diplopoda) distribution: A review. Soil Organisms, 81, 565–597.
Hall, T. (2013) BioEdit v7.2.5. Available from: http://www.mbio.ncsu.edu/bioedit/page2.html (Accessed 12 Nov. 2013)
Hoffman, R.L. (1980) Classification of the Diplopoda. Muséum d´Histoire Naturelle, Genève, 238 pp.
Hoy Jensen, L., Enghoff, H., Frydenberg, J. & Parker, E.D. (2002) Genetic diversity and the phylogeography of parthenogenesis: comparing bisexual and thelytokous populations of Nemasoma varicorne (Diplopoda: Nemasomatidae) in Denmark. Hereditas, 136, 184–194.
http://dx.doi.org/10.1034/j.1601-5223.2002.1360302.xKázmér, M. (1990) Birth, life and death of the Pannonian Lake. Palaeogeography, Palaeoclimatology, Palaeoecology, 79, 171–188.
http://dx.doi.org/10.1016/0031-0182(90)90111-jKimura, M. (1980) A simple method for estimating evolutionary rate of base substitutions through comparative studies of nucleotide sequences. Journal of Molecular Evolution, 16, 111–120.
http://dx.doi.org/10.1007/bf01731581Korsós, Z. (1994) Checklist, preliminary distribution maps, and bibliography of millipedes in Hungary (Diplopoda). Miscellanea Zoologica Hungarica, 9, 29–82.
Korsós, Z. (2000) Rare and endemic millipedes (Diplopoda) of the Abaliget Cave. In: Mock, A., Kováč, L. & Fulín, M. (Eds.), Fauna jaskýň (Cave Fauna). East Slovak Museum, Košice, pp. 61–66.
Košel, V. (2012) Subterranean fauna of the Western Carpathians. Tribun EU, Brno, 210 pp.
Kurnik, I. (1988) Zur Taxonomie ostalpiner Chordeumatida: Vulvenmorphologie und Identifikation der Weibchen. Zoologische Jahrbücher, Abteilung für Systematik, Ökologie und Geographie der Tiere, 115, 229–302.
Kurnik, I. & Thaler, K. (1985) Die Vulven der Chordeumatida: Merkmale von taxonomischer Bedeutung (Diplopoda, Helminthomorpha). Bijdragen tot de Dierkunde, 55, 116–124.
Librado, P. & Rozas, J. (2009) DnaSP v5: A software for comprehensive analysis of DNA polymorphism data. Bioinformatics, 25, 1451–1452.
http://dx.doi.org/10.1093/bioinformatics/btp187Leach, W.E. (1815) A tabular view of the external characters of four classes of animals, which Linné arranged under Insecta; with the distribution of the genera composing three of these classes into orders, &c. and descriptions of several new genera. Transactions of the Linnean Society of London, 11, 306–400.
http://dx.doi.org/10.1111/j.1096-3642.1813.tb00065.xLoksa, I. (1961) A Kovácsi-hegy izéltlábúiról (Die Arthropoden des Kovácsi Berges). Állatani Közlemények, 46, 65–80.
Mahunka, S., Murányi, D. & Kontchán, J. (2013) The role of the Balkan Peninsula in the origin and genesis of the soil fauna of the Carpathian Basin: history, aims and results. Opuscula Zoologica Budapest, 44 (Supplement 1), 5–10.
Mauriès, J.-P., Golovatch, S.I. & Stoev, P. (1997) The millipedes of Albania: recent data, new taxa; systematical, nomenclatural and faunistical review (Myriapoda, Diplopoda). Zoosystema, 19 (2–3), 255–292.
Mikhaljova, E.V. (2004) The millipedes (Diplopoda) of the Asian part of Russia. Pensoft, Sofia-Moscow, 292 pp.
Minelli, A. & Michalik, P. (2015) Diplopoda – reproduction. In: Minelli, A. (Ed.), Treatise of Zoology – Anatomy, Taxonomy, Biology. The Myriapoda, volume 2. Leiden, Koninklijke Brill NV, pp. 237–265.
http://dx.doi.org/10.1163/9789004188273_011Moritz, M. & Fischer, S.-C. (1978) Die Typen der Myriapoden-Sammlung des Zoologischen Museums Berlin. I. Diplopoda. Teil 6: Nachtrag zu den Teilen 1 bis 4. Mitteilungen aus dem Zoologischen Museum in Berlin, 54 (2), 333–343.
http://dx.doi.org/10.1002/mmnz.4830540207Nei, M. & Kumar, S. (2000) Molecular Evolution and Phylogenetics. Oxford University Press, Oxford, 333 pp.
Nyirő, G., Oravecz, O. & Márialigeti, K. (2002) Detection of Wolbachia pipientis infection in arthropods in Hungary. European Journal of Soil Biology, 38 (1), 63–66.
http://dx.doi.org/10.1016/s1164-5563(01)01127-xPapáč, V., Hudec, I., Kováč, Ľ., Ľuptáčik, P. & Mock, A. (2014) Bezstavovce (Invertebrates of the Domica Cave). In: Gaál, L. & Gruber, P. (Eds.), Jaskynný systém Domica – Baradla, jaskyňa, ktorá nás spája. Aggtelek Nemzeti Park, Jósvafő, pp. 269–279.
Ratnasingham, S. & Hebert, P.D.N. (2007) The Barcode of Life Data System. Molecular Ecology Notes, 7, 355–364.
http://dx.doi.org/10.1111/j.1471-8286.2007.01678.xRothenbühler, H. (1900) Ein Beitrag zur Kenntnis der Myriapodenfauna der Schweitz. Revue suisse de Zoologie, 8, 167–192.
http://dx.doi.org/10.5962/bhl.part.17668Saitou, N. & Nei, M. (1987) The neighbor-joining method: A new method for reconstructing phylogenetic trees. Molecular Biology and Evolution, 4, 406–425.
Shear, W.A. (2000) On the millipede family Heterochordeumatidae, with comments on the higher classification of the order Chordeumatida (Diplopoda). Invertebrate Taxonomy, 14, 363–376.
http://dx.doi.org/10.1071/it99016Shear, W. (2011) Class Diplopoda de Blainville in Gervais, 1844. In: Zhang, Z.-Q. (Ed.) Animal biodiversity: An outline of higher-level classification and survey of taxonomic richness. Zootaxa, 3148, 159–164.
Shelley, R.M. (1993) The millipede genus Underwoodia (Chordeumatida: Caseyidae). Canadian Journal of Zoology, 71, 168–176.
http://dx.doi.org/10.1139/z93-023Shelley, R.M. (2002) A revised, annotated, family-level classification of the Diplopoda. Arthropoda Selecta, 11, 187–207.
Silvestri, F. (1898) Contributo alla conosceza dei Chilopodi e Diplopodi della Sicilia. Bollettino della Società entomologica Italiana, 29 (4), 233–261.
Simon, C., Frati, F., Beckenbach, A., Crespi, B., Liu, H. & Flook, P. (1994) Evolution, weighting, and phylogenetic utility of mitochondrial gene sequences and compilation of conserved polymerase chain reaction primers. Annals of the Entomological Society of America, 87, 651–701.
http://dx.doi.org/10.1093/aesa/87.6.651Skoumalová, I. (2010) Mnohonožky Českého a Moravského krasu (Diplopods of Czech and Moravian karst). Diploma thesis. Faculty of Education, Charles University, Prague, 356 pp. Available from: https://is.cuni.cz/webapps/zzp/detail/86982?lang=en (accessed 2 May 2016)
Spelda, J. (2016) Hungarosoma bokori. In: Roskov, Y., Abucay, L., Orrell, T., Nicoson, D., Flann, C., Bailly, N., Kirk, P., Bourgoin, T., DeWalt, R.E., Decock, W. & De Wever, A. (Eds.), Species 2000 & ITIS Catalogue of Life, 2016 Annual Checklist. Available from: www.catalogueoflife.org/annual-checklist/2016/. Species 2000 (accessed 25 July 2016)
Spelda, J., Reip, H.S., Oliveira-Biener, U. & Melzer, R.R. (2011) Barcoding Fauna Bavarica: Myriapoda – a contribution to DNA sequence-based identification of centipedes and millipedes. Zookeys, 156, 123–139.
Strasser, K. (1937) Verhoeffeuma n. g., südostalpiner Höhlendiplopoden. Zoologischer Anzeiger, 118, 225–234.
Szederkényi, T., Kovács, S., Haas, J. & Nagymarosy, A. (2013) Geology and History of Evolution of the ALCAPA Mega-Unit. In: Haas, J. (Ed.), Geology of Hungary. Springer- Verlag, Berlin–Heidelberg, pp. 1–102.
http://dx.doi.org/10.1007/978-3-642-21910-8_1Tamura, K., Stecher, G., Peterson, D., Filipski, A. & Kumar, S. (2013) MEGA6: Molecular Evolutionary Genetics Analysis Version 6.0. Molecular Biology and Evolution, 30, 2725–2729.
http://dx.doi.org/10.1093/molbev/mst197Verhoeff, K.W. (1899) Zur vergleichenden Morphologie, Phylogenie, Gruppen- und Art-Systematik der Chordeumiden. Archiv für Naturgeschichte, 65 (1), 95–154.
Verhoeff, K.W. (1901) Diplopoden aus Herzegowina, Ungarn und Baiern. Archiv für Naturgeschichte, 67, 221–240.
Verhoeff, K.W. (1911) Über Brachychaeteuma n. g. und Titanosoma jurassicum aus England. Zoologischer Anzeiger, 38, 455–458.
Verhoeff, K.W. (1913) Über Diplopoden. 63. Aufsatz: Syngonopodium n. g. Sitzungsberichte der Gesellschaft Naturforschender Freunde zu Berlin, 204, 269–280.
Verhoeff, K.W. (1914) Über Diplopoden. 69. Aufsatz: Ascospermophoren aus Japan. Zoologischer Anzeiger, 43, 342–370.
Verhoeff, K.W. (1929) Arthropoden aus Südostalpine Höhlen, Heft 7. Mitteilungen über Höhlen- und Karstforschung, 1–21.
Verhoeff, K.W. (1928) Zur Kenntnis der Diplopodenfauna Ungarns. 109. Diplopoden-Aufsatz (Chilopoden). Állatani Közlemények, 25, 124–126; 182–199.
Verhoeff, K.W. (1932) Dr. Bronn, H.G.’s Klassen und Ordnungen des Tier-Reichs wissenschaftlich dargestellt in Wort und Bild. 5 (II). Gliederfüssler: Arthropoda. Klasse Diplopoda, 2. Akademische Verlagsgesellschaft m.b.H., Leipzig, pp. 1073–2084.
Werren J.H. & Windsor D.M. (2000) Wolbachia infection frequencies in insects: evidence of a global equilibrium? Proceedings of the Royal Society, B-Biological Sciences, 267 (1450), 1277–1285.
http://dx.doi.org/10.1098/rspb.2000.1139Witzel, K.-P., Zakharov, I.A., Goryacheva, I.I., Adis, J. & Golovatch, S.I. (2003) Two parthenogenetic millipede species/lines of the genus Poratia Cook & Cook, 1894 (Diplopoda, Polydesmida, Pyrgodesmidae) found free from Wolbachia bacteria. African Invertebrates, 44, 331–338.