ZooKeys 408: 8 fe) 05 (20 | 4) A peer-revi iewed open-access journa I doi: 10.3897/zookeys.408.6612 RESEARCH ARTICLE #ZooKey WWW.ZOO keys.o rg Launched to accelerate biodiversity research New tardigrade records for the Baltic states with a description of Minibiotus formosus sp. n. (Eutardigrada, Macrobiotidae) Krzysztof Zawierucha'', Jakub Dziamiecki'*, Natalia Jakubowska’4, Lukasz Michalczyk?', Lukasz Kaczmarek'4 | Department of Animal Taxonomy and Ecology, Faculty of Biology, A. Mickiewicz University, Umultowska 89, 61-614 Poznan, Poland 2 Department of Water Protection, Faculty of Biology, A. Mickiewicz University, Umultowska 89, 61-614 Poznan, Poland 3 Department of Entomology, Institute of Zoology, Jagiellonian University, Gronostajowa 9, 30-387 Krakow, Poland T /ttp-//zoobank. org/0404754A-B7C5-4D31-95B9-653EB2233DFD ¢ Attp://zoobank. org/46066F30-A5ED-45D9-93DF-04C22F5888D 1 § Attp://zoobank. org/354A06C9-30FE-45DD-8BEE-3D1046F3DD86 | Attp-//zoobank. org/6592787E-AAB 1-4717-90C9-B967488FO6F4 Ql 2ttp-//zoobank. org/FE20F8A0-B5E8-4C17-869C-29AA12DFF672 Corresponding author: Lukasz Kaczmarek (kaczmar@amu.edu.pl) Academic editor: L. Penev | Received 14 November 2013 | Accepted 14 March 2014 | Published 13 May 2014 http://z00bank.ore/D5A553D7-5B78-4304-8 97 1-6AOCF73FC698 Citation: Zawierucha K, Dziamiecki J, Jakubowska N, Michalczyk £, Kaczmarek £ (2014) New tardigrade records for the Baltic states with a description of Minibiotus formosus sp. n. (Eutardigrada, Macrobiotidae). ZooKeys 408: 81-105. doi: 10.3897/zookeys.408.6612 Abstract In sixteen moss, lichen and mixed (moss/lichen) samples, collected from Estonia, Latvia and Lithuania, 291 specimens, 48 simplexes, including one exuvium with 6 eggs, and 8 free-laid eggs of eutardigrades were found. In total, 17 species, together with one new to science, were identified (all are new records for the Baltic states): Astatumen bartosi, Diphascon (Adropion) prorsirostre, D. (Diphascon) bullatum, D. (D.) pingue pingue, D. (D.) recamieri, D. (D.) rugosum, Hypsibius convergens, H. dujardini, H. cf. scabro- pygus, Isohypsibius ronsisvallei, I. sattleri, Macrobiotus harmsworthi harmsworthi, M. hufelandi hufelandi, Milnesium asiaticum, Milnesium tardigradum tardigradum, Minibiotus formosus sp. n. and Paramacrobiotus richtersi. The new species is most similar to Minibiotus gumersindoi, but differs from it mainly by the pres- ence of two types of cuticular pores, the absence of a triangular or pentagonal arrangement of pores above a single large pore on legs, the presence of granulation on all legs and a different macroplacoid length sequence. In this paper we also provide photographs and morphometrics of H. cf. scabropygus. Copyright K. Zawierucha et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 82 Krzysztof Zawierucha et al. / ZooKeys 408: 81-105 (2014) Keywords Estonia, Europe, Hypsibius cf. scabropygus, Latvia, Lithuania, new species, Tardigrada Introduction The Baltic States, i.e. Estonia, Latvia and Lithuania, are located on the eastern coast of the Baltic Sea, and fall within the Palearctic ecozone (Holt et al. 2012). The topogra- phy of the three countries is dominated by lowlands with the highest peaks at ca. 300 m asl. The temperate climate is intermediate between maritime and continental. Even though the phylum Tardigrada is cosmopolitan and currently comprises ca. 1,200 species (Degma et al. 2013), so far only six tardigrade taxa have been reported from the Baltic States. Specifically, two from Estonia: Eremobiotus alicatai (Binda, 1969) and Isohypsibius cf. marcellinoi (Binda & Pilato, 1971), two from Latvia: Paramacro- biotus richtersi group and Macrobiotus hufelandi group and two from Lithuania: Mac- robiotus sp. and Ramazzottius sp. (Satkauskiené and Vosyliiite 2010, Zawierucha and Kazmierski 2012, Ziemelis et al. 2012). In this study we report seventeen tardigrade species, which are all new records for the Baltic States. Moreover, one of these species is also new to science. The new spe- cies belongs to the genus Minibiotus R.O. Schuster, 1980, that until 1988 contained only a single species, Minibiotus intermedius (Plate, 1888). In 1988 Pilato and Clax- ton (1988) described Minibiotus maculartus, and within the last decade fourteen new Minibiotus species have been described. Several species have also been transferred to Minibiotus from the genus Macrobiotus based on characters defined by Claxton (1998) and later supplemented by Guidetti et al. (2007) (Michalczyk and Kaczmarek 2003a, Pilato et al. 2003, Michalczyk and Kaczmarek 2004, Guil and Guidetti 2005, Michal- czyk et al. 2005, Pilato and Lisi 2006b, Li et al. 2008, Fontoura et al. 2009a, b, Rossi et al. 2009, Meyer and Hinton 2009, Meyer and Domingue 2011, Meyer et al. 2011 Meyer 2012). Currently, the total number of Minibiotus species amounts to as many as forty seven. In addition to the description of the new species, we also provide morphometric data and photographs of Hypsibius cf. scabropygus, a rare species that belongs to a large group of hypsibiids with at least partially sculptured dorsal cuticle and pharynx with two macroplacoids and without the microplacoid. Material and methods Sixteen moss, lichen and mixed (moss/lichen) samples from trees, soil and stones were collected from 15 localities in Estonia, Latvia and Lithuania between the 29 April and the 5 May 2012 by the third author (more details below). Samples were collected and examined for tardigrades using standard methods (see Dastych 1980). After extraction, Tardigrades from the Baltic States 83 animals were mounted on microscope slides in Hoyer’s medium. All specimens were examined measured and photographed using Phase Contrast Microscopy (PCM) or Scanning Electron Microscopy (SEM). In total 358 specimens (including 47 simplexes), one exuvium with 6 eggs, and 8 free-laid eggs were examined. All measurements are given in micrometers [um]. Structures were measured only if their orientation was appropriate. Body length was measured from the anterior extrem- ity to the end of the body, excluding the hind legs. Buccal tube length and the level of the stylet support insertion point were measured according to Pilato (1981). Buccal tube width was measured as the external diameter at the level of the stylet support inser- tion point. Lengths of the claw branches were measured from the base of the claw to the top of the branch including accessory points for Minibiotus and according to Beasley et al. (2008) for Hypsibius. The pt ratio is the ratio of the length of a given structure to the length of the buccal tube expressed as a percentage (Pilato 1981). Macroplacoid length sequence is given according to Kaczmarek et al. (2014b), i.e. macroplacoids are listed from the shortest to the longest and their relative sizes are denoted with appropriate inequality, approximation and/or equality signs (<, <, ~, =). Morphometric data were handled using the ‘Macrobiotoidea’ ver. 1.1 template available from the Tardigrada Register (www.tardigrada.net/register, Michalczyk and Kaczmarek 2013). For species identification and differentiation, keys in Claxton (1998), Fontoura and Pilato (2007), Kaczmarek et al. (2011), Michalczyk et al. 2012a, b and Ramazzotti and Maucci (1983), and original descriptions and redescriptions (Ehrenberg 1859, Ramaz- zotti 1959, 1962, Horning et al. 1978, Bertolani and Rebecchi 1993, Dastych 1988, 1990, Binda and Pilato 1992, Michalczyk and Kaczmarek 2004, Michalczyk et al. 2005, Miller et al. 2005, Fontoura et al. 2009a, b, Meyer and Hinton 2009, Meyer et al. 2011) as well as for insertion of the stylet muscles Pilato (2013) were used. Tardigrade taxonomy is presented according to Marley et al. (2011). Only specimens determined to species level are provided in the list of species (we omitted all specimens determined only to the species group level, e.g. the /ufelandi group or the oberhaeuseri group). In the species list Roman numbers indicate sample codes (see sampling localities) and Arabic numbers indicate the number of specimens, exuvia/simplexes and eggs. Raw data underlying the description of Minibiotus formosus sp. n. are deposited in the Tardigrada Register (Michalczyk and Kaczmarek 2013) under www.tardigrada. net/register/0012.htm. Sampling localities I. 56°03'08"N; 24°24'10"E, ca. 33 m asl: Lithuania, Panevézys county, Pasvalys district municipality, along the road E67, 0.5 km before the turning to Pas- valys, moss from tree and soil (slide code: LT 2422), date: 29.04.2012. Ik 55°25'59"N; 24°13'32"E, ca. 59 m asl: Lithuania, Kaunas county, Kédainiai district municipality, Truskava city, near a church, lichens from tree and moss from soil (slide code: LT 2423), date: 29.04.2012. 84 Il. IV. VI. VIL. Vie IX. XI. XII. XIII. XIV. Krzysztof Zawierucha et al. / ZooKeys 408: 81-105 (2014) 55°17'12"N; 23°58'57"E, ca. 30 m asl: Lithuania, Kaunas county, Kédainiai district municipality, Kédainiai city, Kranto II street; moss from wall (slide code: LT 2424), date: 29.04.2012. 55°17'13"N; 23°58'56"E, ca. 30 m asl: Lithuania, Kaunas county, Kédainiai district municipality, Kédainiai city, Paeismilgio street; moss from stone (slide code: LT 2425), date: 29.04.2012. 55°43'35"N; 24°21'30"E, ca. 62 masl: Lithuania, Panevézys county, Panevézys district municipality, Panevézys city, Garden Street near Holy Trinity Rector; moss from tree (slide code: LT 2440), date: 05.05.2012. 56°38'53"N; 23°43'18"E, ca. 7 m asl: Latvia, Zemgale region, Jelgava mu- nicipality, Jelgava city, City Park; moss from soil (slide code: LO 2426), date: 29.04.2012. 57°10'33"N; 24°50'32"E, ca. 45 m asl: Latvia, Vidzeme region, Sigulda mu- nicipality, Gutmana Cave in the Gauja National Park; moss from rocks (slide code: LO 2427), date: 30.04.2012. 56°23'55"N; 24°07'33"E, ca. 25 m asl: Latvia, Zemgale region, Bauska mu- nicipality, along Road No P103, 0.5 km from Saulaine; lichens from tree (slide code: LO 2428) date: 29.04.2012. 57°09'55"N; 24°51'03"E, ca. 73 m asl: Latvia, Vidzeme region, Sigulda mu- nicipality, Turaida city, Turaida Castle; moss from stone (slide code: LO 2430), date: 30.04.2012. 56°54'32"N; 24°08'45"E, ca. 10 m asl: Latvia, Riga Region, boundary of Kekava municipality, along road no A2; moss from tree (slide code: LO 2431), date: 30.04.2012. 57°09'59"N; 24°50'59"E, ca. 91 m asl: Latvia, Vidzeme region, Sigulda mu- nicipality, Sigulda city, Sigulda Castle; moss from stone (slide code: LO 2432), date: 30.04.2012. 56°41'22"N; 23°47'43"E, ca. 4 m asl: Latvia, Zemgale region, Ozolnieki mu- nicipality, Ozolnieki city, about 100 m from the Ozolnieki Lake; moss from soil (slide code: LO 2433), date: 29.04.2012. 57°45'43"N; 24°20'59"E, ca. 3 m asl: Latvia, Vidzeme region, Salacgriva municipality, Salacgriva city; moss from soil, near the beach (slide code: LO 2434), date: 01.05.2012. 59°10'44"N; 24°30'06"E, ca. 59 m asl: Republic of Estonia, Harju county, Kernu Parish municipality, Road No 4, moss from tree (slide code: ES 2420), date: 04.05.2012. 59°10'44"N; 24°30'06"E, ca. 59 m asl: Republic of Estonia, Harju county, Kernu Parish municipality, Road No 4, moss from tree (slide code: ES 2421), date: 04.05.2012. 58°48'47"N; 24°24'46"E, ca. 32 m asl: Republic of Estonia, Rapla County, Marjamaa municipality, forest near Konuvere village, moss from tree (slide codes: ES 2487), date: 29.04.2012. Tardigrades from the Baltic States 85 Results Taxonomic accounts of species found in the study Phylum: Tardigrada (Spallanzani, 1777) Class: Eutardigrada Richters, 1926 Order: Apochela Schuster, Nelson, Grigarick and Christenberry, 1980 Family: Milnesiidae Ramazzotti, 1962 Genus: Milnesium Doyére, 1840 Milnesium asiaticum Tamanov, 2006 Localities and specimen numbers. XV: | specimen. Remarks. Our specimen corresponds perfectly to the original description. Milne- sium asiaticum was originally described from Kirghizstan and subsequently found in the Svalbard archipelago (Tumanov 2006, Kaczmarek et al. 2012). Milnesium tardigradum tardigradum Doyére, 1840 Localities and specimen numbers. VIII: 31 specimens (including 6 simplexes) + 1 exuvium with 6 eggs. Remarks. Specimens correspond perfectly with the redescription by Michalczyk et al. (2012a, b). This species was reported from many localities throughout the World, however records prior to Michalczyk et al. (2012a, b) need to be verified. So far, all confirmed localities are exclusively European (Michalczyk et al. 2012a, b). Order: Parachela Schuster, Nelson, Grigarick & Christensen, 1980 Superfamily: Hypsibioidea Pilato, 1969 (in Marley et al. 2011) Family: Hypsibiidae Pilato, 1969 Subfamily: Diphasconinae Dastych, 1992 Genus: Diphascon Plate, 1888 Subgenus: Diphascon (Diphascon) Pilato, 1987 Diphascon (Diphascon) bullatum Murray, 1905 Localities and specimen numbers. XIII: 1 specimen. Remarks. Specimens correspond well with the limited original description (Murray 1905) and also with later descriptions (Argue 1974 and Dastych 1980, 1988). This spe- cies is very similar to D. (D.) patanei (Binda & Pilato, 1971) and, as suggested by Dastych (1988), these two species could be synonymous. Thus, to clarify the taxonomic status of D. (D.) patanei, a re-description of D. (D.) bullatum is necessary based on material from locus typicus in Scotland (the type material probably does not exist) (Dastych 1988). 86 Krzysztof Zawierucha et al. | ZooKeys 408: 81-105 (2014) Diphascon (Diphascon) pingue pingue (Marcus, 1936) Localities and specimen numbers. XV: 11 specimens. Remarks. Although we have found only 11 specimens, we were confident in iden- tifying them to D. (D.) pingue because they corresponded perfectly to the partial re- descriptions by Pilato and Binda (1997/1998, 1999) and we also identified them with the key by Fontoura and Pilato (2007). The species belongs to the pingue group and has been previously recorded from numerous localities throughout the World, how- ever the majority of records should be verified based on the modern taxonomy (Pilato and Binda 1997/1998, 1999). Currently, exclusively verified localities of this species are only from Europe and North America (Pilato and Binda 1997/1998). Diphascon (Diphascon) recamieri Richters, 1911 Localities and specimen numbers. XV: | specimen. Remarks. The species has previously been found in many localities, mostly in the Holarctic (McInnes 1994). Diphascon (Diphascon) rugosum (Bartos, 1935) Localities and specimen numbers. II: 3 specimens. Remarks. ‘The species has previously been found in many localities in the Holarctic (McInnes 1994). Genus: Diphascon Pilato, 1987 Subgenus Diphascon (Adropion) prorsirostre Thulin, 1928 Localities and specimen numbers. XIV: 2 specimens, XV: 2 specimens. Remarks. The species has been previously found in many localities, mostly in the Holarctic (McInnes 1994). Subfamily: Hypsibiinae Pilato, 1969 Genus: Hypsibius Ehrenberg, 1848 Hypsibius convergens (Urbanowicz, 1925) Localities and specimen numbers. [X: 6 specimens (including 2 simplexes). Remarks. Belonging to the cosmopolitan convergens-dujardini complex of species (McInnes 1994, Miller et al. 2005, Kaczmarek and Michalczyk 2009a, Kaczmarek et al. Tardigrades from the Baltic States 87 2014a), H. convergens used to be considered cosmopolitan, but it is most likely a complex of very similar (possibly also cryptic) species found throughout the world. ‘The original H. convergens description no longer conforms to modern standards and therefore a rede- scription is required. Nevertheless, the examined specimens correspond perfectly with the original description and the H. convergens characteristics reviewed by Miller et al. (2005). Hypsibius dujardini (Doyere, 1840) Localities and specimen numbers. XIV: 2 specimens (including 1 simplex), XV: 5 specimens. Remarks. H. dujardini belongs to the cosmopolitan convergens-dujardini complex of species (McInnes 1994, Miller et al. 2005, Kaczmarek and Michalczyk 2009a, Kac- zmarek et al. 2014a), and used to be considered cosmopolitan, but it is most likely a complex of similar (possibly cryptic) species found throughout the world. Due to the limited original description, H. dujardini needs a modern redescription. Nevertheless, the examined specimens correspond perfectly with the original description and the H. dujardini characteristics reviewed by Miller et al. (2005). Hypsibius cf. scabropygus Cuénot, 1929 Table 1, Figs 1-7 Localities and specimen numbers. XI: 1 simplex, XII: 1 specimen, XIV: 34 specimens (including 4 simplexes), XV: 24 specimens (including 6 simplexes). Description (measurements in Table 1). Adults. Body transparent/white (af- ter preparation), eyes present in 6 of 15 (40%) specimens mounted in Hoyer’s me- dium (Fig. 1). Dorsal cuticle sculptured: from head to legs If without tubercles but clearly thickened, from legs II to the caudal end of the body (including legs IV) with irregular tubercles and platelets. ‘Tubercles increasing in size from the anterior to the posterior part of the body, reaching maximum dimensions between legs III and IV, where tubercles sometimes merge and form irregular platelets (Figs 2—5). Irregular tubercles 1.0—6.0 um in diameter. Ventral cuticle smooth (i.e. without sculpturing). Gibbosities and cuticular pores absent. Bucco-pharyngeal apparatus of the Hypsibius type, without the ventral lamina, and with forked apophyses for stylet muscles (Fig. 6). Peribuccal lamellae absent. Teeth in the oral cavity armature absent or not visible under PCM. Pharyngeal bulb with apophyses and with two granular macroplacoids (both, without constrictions). Macro- placoid length sequence 2<1. Microplacoid and septulum absent. Claws of the Hypsibius type, internal claws much smaller and of a different shape than the external claws (Fig. 7). All main branches with large accessory points. Smooth, indistinct areoles under claws usually visible only on posterior claws IV. Cuticular bars under claws I-III absent but a small bar is present near the posterior claw IV (Fig. 7, arrow). Eggs. Unknown. 88 Krzysztof Zawierucha et al. / ZooKeys 408: 81-105 (2014) ih! } x . # a Pa m Figures 1-5. Hypsibius cf. scabropygus Cuénot, 1929: | habitus (dorso-lateral view) 2—4 caudo-dorsal cuticle with distinct sculpturing — tubercles and tubercles merged into platelets 5 a single caudo-dorsal platelet. (1-3: PCM, 4-5: SEM). Remarks. Hypsibius scabropygus has been recorded from many localities, mostly in the Holarctic (McInnes 1994). In general, our specimens correspond to the original de- scription by Cuénot (1929, 1932) and later descriptions by Marcus (1930) (=Hypsibius callimerus spec. nov.), and by Ramazzotti and Maucci (1983). However, importantly, none of the above mentioned descriptions reported a bar between anterior and posterior claws IV, which is present in all our specimens. Given the bar is small, it is possible that it was overlooked by Cuénot and later authors. If, however, H. scabropygus does not have the bar, then our specimens should probably be classified as a new species. Thus, until Tardigrades from the Baltic States 89 Figures 6-7. Hypsibius cf. scabropygus Cuénot, 1929: 6 bucco-pharyngeal apparatus (dorso-ventral pro- jection, ventral placoids in the insert) 7 claws IV (arrow indicates a small cuticular bar near the posterior claw). (Both PCM). H. scabropygus is redescribed, our Latvian and Estonian records should be regarded as 7. cf. scabropygus. As there is a possibility of our specimens belonging to a new species, we provide standard morphometrics (Table 1) and photographs (Figs 1-7). Subfamily: Itaquasconinae Rudescu, 1964 Genus: Astatumen Pilato, 1997 Astatumen bartosi (Weglarska, 1959) Localities and specimen numbers. XIV: 1 specimen. Remarks. Our specimen corresponds perfectly with characters of A. bartosi pro- posed by Dastych (1988) with the main difference between A. bartosi and A. trinacriae being the absence/presence of cuticular bars on legs II-III. Due to the notorious dif- ficulties in differentiating the two species, the actual distribution of A. bartosi cannot currently be described with confidence. McInnes (1994) cited this species from several localities in Europe and from single African, Asian and South American sites. Superfamily: Isohypsibioidea Marley, McInnes & Sands, 2011 Family: Isohypsibiidae Marley, McInnes & Sands, 2011 Genus: Isohypsibius Thulin, 1928 Isohypsibius ronsisvallei Binda & Pilato, 1969 Localities and specimen numbers. III: 1 specimen. Remarks. ‘The species has previously been reported from several, mostly European, localities in the Holarctic (McInnes 1994). 90 Krzysztof Zawierucha et al. / ZooKeys 408: 81-105 (2014) Table 1. Measurements and pt values of selected morphological structures of Hypsibius cf. scabropygus Cuénot, 1929 mounted in Hoyer’s medium (N — number of specimens/structures measured, RANGE refers to the smallest and the largest structure among all measured specimens; SD — standard deviation). CHARACTER N RANGE MeN av pt um | pt | um | pt Body length LTB ZN 239" 9294) SO elee se Buccopharyngeal tube Buccal tube length 24.6} — | 2.0) —- Stylet support insertion point SAE | T3535 4e95 Pale |. 2G. Buccal tube external width Pe alee LBy | ee | O28 POS: Buccal tube internal width =| BE | 0:7 | 2H | Ode | OF Placoid lengths Ele Macroplacoid 1 Saez 2.4 | 9.6 | 0.4} 1.3 Macroplacoid 2 SRR 8.9103] LI Macroplacoid row 2635)|| 333" | 202 |- OS? | «8 Claw 1 lengths External base Dada 6505 aoa) Ore | ai2sd External primary branch 3652. | "6.9 | 28/24 FL? | OO External secondary branch 24.4| 44 |17.9| 10] 3.9 Internal base Paap 13.4| 0.6 | 1.8 Internal primary branch Ea ea si 4.8 | 19.3| 0.6 | 2.0 Internal secondary branch = | 765./33-| 1 S006! |) 2:0 Claw 2 lengths Saat External base | ir ore:| are L630) Wa75,| 22 External primary branch 8.5 | 34.7| 1.3 | 44 External secondary branch 548 | 290.7 | 256: Internal base 14.6| 0.7 | 2.2 Internal primary branch 22:0: | 22 Internal secondary branch L67)| 095) G0 Claw 3 lengths na | External base | 238) Base | LAD.) ShOe | EBay External primary branch ea 8.8 | 35.7| 1.1 | 44 External secondary branch ena | 5.2.5) 2EO| 10. | 4.3 Internal base EIR AEr4 LSD: SOG: ISD Internal primary branch 2/.3)| HE | BEG OD. | Bel Internal secondary branch 24:8"| 3:9: | 1650) 09°) 3:5 Claw 4 lengths Anterior base 20.1| 4.1 | 16.8} 0.6 | 2.2 Anterior primary branch oP Renee ah ee Anterior secondary branch = | 479 | 4.88 | TO. 94) 2. | OZ Posterior base aps, Aes | tae COrS | Bab Posterior primary branch | — | 60.6 | 10.3 | AID. | 29) | 1h2 Posterior secondary branch Sa ea 2) eid. |e Sal) ee Tardigrades from the Baltic States eH Isohypsibius sattleri (Richters, 1902) Localities and specimen numbers. [X: 1 specimen, XI: 8 specimens (including 5 simplexes), XIV: 3 specimens, XV: 5 specimens, XVI: 1 specimen. Remarks. ‘The species has previously been reported from many localities throughout the World, thus it is considered cosmopolitan (McInnes 1994, Kaczmarek et al. 2014a). Superfamily: Macrobiotoidea Thulin, 1928 in Marley et al. 2011 Family: Macrobiotidae Thulin, 1928 Genus: Macrobiotus C.A.S. Schultze, 1834 Macrobiotus harmsworthi harmsworthi Murray, 1907 Localities and specimen numbers. XI: 1 egg, XIV: 8 specimens, 1 egg. Remarks. ‘The species belongs to the harmsworthi group which is widely distributed across a broad range of ecosystems throughout the world (McInnes 1994, Kaczmarek et al. 2014a). In the last decade many new species within this group were described from a variety of localities (Michalczyk and Kaczmarek 2003b, Pilato et al. 2004, Tumanov 2005a, Pilato and Lisi 2006a, b, Pilato et al. 2006a, Kaczmarek et al. 2007, Kaczmarek and Michalczyk 2009b, Pilato and Lisi 2009a, Rossi et al. 2009, see also Kaczmarek et al. 2011 for the diagnostic key to the group). Due to many uncertain reports of M. harmsworthi harmsworthi, especially in older literature, the distribution of the species is currently unknown. Specimens found in the present study correspond well to the characters presented in Pilato et al. (2000) and were successfully identified with the key by Kaczmarek et al. (2011). Macrobiotus hufelandi hufelandi C.A.S. Schultze, 1833 Localities and specimen numbers. XIV: 4 specimens, | egg. Remarks. ‘The species belongs to the /ufelandi group which is widely distributed across a broad range of ecosystems throughout the world (McInnes 1994, Kaczmarek et al. 2014a). In the last decade new species belonging to this group have been described from various localities (Pilato et al. 2003, Kaczmarek and Michalczyk 2004, Dastych 2002, 2005, Fontoura et al. 2008, Bartels et al. 2009, Kaczmarek and Michalczyk 2009b, Pilato and Lisi 2009b, Bertolani et al. 2011, Biserov et al. 2011, Pilato et al. 2012, Guidetti et al. 2013, see also Bertolani and Rebecchi 1993 for the diagnostic key to the group). Due to many uncertain reports of M. hufelandi hufelandi, especially in older literature, the distribution of the species is currently unknown. Specimens found in the present study correspond well with the redescription by Bertolani and Rebecchi (1993). 92 Krzysztof Zawierucha et al. / ZooKeys 408: 81-105 (2014) Genus: Minibiotus R.O. Schuster, 1980 Minibiotus formosus sp. n. http://zoobank.org/BDBE49B7-84CF-4FE2-BE55-A399A537DE77 http://species-id.net/wiki/Minibiotus_formosus http://www.tardigrada.net/register/0012.htm Tables 2—3, Figs 8-15 Type material. Holotype and 23 paratypes, 24 specimens (including 2 simplexes) and 3 unembryonated eggs). Type locality. 57°10'33"N; 24°50'32"E, ca. 45 m asl: Latvia, Vidzeme region, Sigulda municipality, Gutmana Cave in the Gauja National Park; moss from rock (1 sample, slide codes: LO 2427/*, where the asterisk can be substituted by any of the following numbers: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12). Description (measurements in Table 2). Body white/colourless (Fig. 8). Eyes pre- sent in 18 of 24 (75%) specimens mounted in Hoyer’s medium. Entire cuticle covered with small (0.4—-1.1 um) and large (1.9-2.5 um) round or oval pores (Fig. 9). Pores arranged in 9-10 poorly defined transverse bands. Pores on the dorsal cuticle arranged more densely than on the ventral cuticle. A single large pore (diameter: 2.1—2.9 um) present on external side of legs I-III (Fig. 12, arrow). A ring of pores around the mouth opening absent. Cuticle without granulation, except for legs which are all covered with fine and regular granulation (better developed on legs IV) visible only in larger specimens (Fig 12, arrowhead). Mouth antero-ventral. Ten peribuccal papulae present. Bucco-pharyngeal ap- paratus of the Minibiotus type (Figs 10-11). Oral cavity armature absent or not visible under PCM. Buccal tube with a poorly visible ventral lamina and with an anterior and a posterior bend (both visible in lateral view only, Fig. 11). Buccal tube walls thickened just below the stylet supports insertion point. Pharyngeal apophyses triangular, very near to the first macroplacoid. Three granular macro- placoids and a minute microplacoid present in the pharyngeal bulb. All macropla- coids of similar but not identical sizes, the macroplacoid length sequence: 2<1<3. Septulum absent. Claws of the Macrobiotus type (Figs 12-13). Primary branches of claws with thin, but obvious accessory points detaching at the apogee of the primary branch curve. Smooth lunules present on all legs, distinctly larger under external and posterior claws. Bars and other cuticular thickenings on legs absent. Eggs (measurements in Table 3). White/transparent, laid freely (Fig. 14). Spheri- cal, without areolation. Processes in the shape of short, smooth, slightly flexible cones (Fig. 15). Processes are distributed on the surface of the egg close one to another but never in contact. Surface between processes smooth under PCM (Fig. 15). Remarks. Since ventral lamina is very poorly visible, the measurements of this structure are not included in Table 2. Three unembryonated eggs have been found alongside the described specimens. Given that M. formosus sp. n. was the only Minibiotus species in the Tardigrades from the Baltic States ) Y) Figures 8—1 1. Minibiotus formosus sp. n.: 8 habitus (holotype, ventral view) 9 dorsal cuticle with pores (holotype) 10-11 bucco-pharyngeal apparatus (10 dorso-ventral projection, paratype I I lateral view, paratype). All PCM. sample and because no Ramazzottius Binda & Pilato, 1986 was found in the samples, we assumed that these eggs belong to the new species. Etymology. Given that we found the composition of small and large pores in the new species beautiful, we decided to name the animal after this impression (in Latin ‘formosus means ‘beautiful’). 94 Krzysztof Zawierucha et al. / ZooKeys 408: 81-105 (2014) Table 2. Measurements and pt values of selected morphological structures of Minibiotus formosus sp. n. mounted in Hoyer’s medium (N — number of specimens/structures measured, RANGE refers to the smallest and the largest structure among all measured specimens; SD — standard deviation, ? — trait ori- ented unsuitably for measurement). Holotype N RANGE SD um |) pt um pt a | [ase | ee] 848 Flea era eee emi =a ey el ae [22 | fm 12 a i ae CHARACTER Body length Buccal tube Length External width 2.0 7.0 Stylet support insertion point] 9 | 9.5 |~ | 12.2 | 49.5 | - | 56.2 | 10.9 | 53.8 | 0.9 | 2.0 | 12.2 | 53.3 Flies Internal width Placoid lengths Macroplacoid 1 Macroplacoid 2 Macroplacoid 3 Microplacoid Macroplacoid row Placoid row -| 9.9 | 1.6 | 7.8 | 7.8 7.4 }9 | 05 |-| 07 | 2.4 [| ee oli nS : 18 | 7.9 = 7.4 : 19 -| 47 | 07 | 3.3 [0.1] 08 | 08 | 3.5 : }9 | 1.4 [- 8 | 0.5 |- I) Le? Die 07 | 3.1 ai eae a 8.3 Do. P28 6.7 |29.3 Claw 1 lengths PAE ea es External primary branch 27.1 External secondary branch ? Internal primary branch 6137275 Internal secondary branch 4.6 | 20.1 Claw 2lengths | | | | | [| | | | | | | | External primary branch DEF | 25.8 External secondary branch Internal primary branch 27.1 Internal secondary branch Claw 3tengths | | | | | | | | | | Tt External primary branch 6.9 | 30.1 External secondary branch 4,9 | 21.4 Internal primary branch 6.4 | 27.9 Internal secondary branch Claw 4 lengths Anterior primary branch Anterior secondary branch Posterior primary branch Posterior secondary branch Type depositories. Holotype 23 paratypes and 3 eggs are deposited in the De- partment of Animal Taxonomy and Ecology at the Adam Mickiewicz University (Poznan, Poland). Tardigrades from the Baltic States oD Figures 12-15. Minibiotus formosus sp. n.: 12 leg II with claws, granulation (arrowhead) and a single large pore (arrow) (holotype) 13 claws IV (paratype) 14 egg (mid-section) 15 egg surface with processes. All PCM. Table 3. Measurements of selected morphological structures of Minibiotus formosus sp. n. eggs mounted in Hoyer’s medium. CHARACTER ege | egg 2 egg 3 Diameter of egg without processes 45.7 44,1 Diameter of egg with processes 55.6 55.1 4 Process height 4.5-5.2 4.85.2 4.6—-5.3 Process base width 2.83.4 2,8-3,1 2.42.6 Process base/height ratio 57%—-69% 54%-65% 47%—-57% Distance between processes 2.0-2.5 1.9-3.9 1.82.0 Number of processes on the egg circumference 30 29) 30 96 Krzysztof Zawierucha et al. / ZooKeys 408: 81-105 (2014) Differential diagnosis The new species is most similar to M. gumersindoi Guil & Guidetti, 2005, but it differs from it by: the presence of two types of cuticular pores (small and large) in the new species vs pores of a uniform size in MZ. gumersindoi, the absence of a triangular or a pentagonal arrangement of pores placed above a single large pore on legs, the presence of granulation on legs, a different macroplacoid length sequence (2<1<3 in the new species vs 1=2=3 in M. gumersindoi), and by slightly larger macroplacoids (I: 1.3-1.9 um; I: 1.2—1.7 um III: 1.4—2.2 um in the new species vs 1.0 um in M. gumersindoi). Other species to which M. formosus sp. n. is similar by some characteristics of adult and/or egg morphology (e.g. pores in transverse bands, eggs with conical processes), include species listed below. The new species differs specifically from: e M. bisoctus (Horning et al. 1978) by: the absence of trilobed and star-shaped pores (although their presence was not mentioned in the original description, they are clearly visible in Fig. 114 in Horning et al. (1978), and by stylet supports in- serted in a more anterior position (pt=49.5—56.2 in the new species vs pt=60.3 in M. bisoctus (according to Claxton 1998)). e M. eichhorni Michalczyk & Kaczmarek, 2004 by: a different arrangement of pores on the dorsal cuticle (9-10 transverse bands in the new species vs 8 bands in M. eichhorni), the absence of star-shaped pores, the absence of four pores around the mouth opening, the presence of a single large pore on lateral sides of legs I-III, slightly shorter buccal tube (17.7—22.9 um in the new species vs 24.7-34.2 um in M. eichhorni), stylet supports inserted in more anterior position (9.5—12.2 um [pt=49.5—56.2] in the new species vs 16.2—23.8 um [pt=65.4-70.6] in M. eich- horni), a different macroplacoid sequence (2<1<3 in the new species vs 2<3<1 um in M. eichhorni), slightly shorter placoids, and by slightly smaller external claws I-IV (compare Table 2 below and Table 1 in Michalczyk and Kaczmarek 2004 for exact differences in dimensions of placoids and claws). e 6M. furcatus (Ehrenberg, 1859) (according to Binda and Pilato 1992) by: the ab- sence of tri- and quadrilobed cuticular pores, the presence of two types of cuticular pores (small and large in the new species vs uniformly small pores present in /. furcatus), the presence of a single large pore on each of legs I-HI, the presence of granulation on legs, the absence of the oral cavity armature, stylet supports in- serted in a more anterior position (pt=49.5—56.2 in the new species vs pt~68.4 in M. furcatus), a different macroplacoid length sequence (2<1<3 in the new species vs 2<3<1 in M. furcatus), and by egg processes without an obvious flexible portion (and never bifurcated). e M. harrylewisi Meyer & Hinton, 2009 by: the absence of tri- and quadrilobed cuticular pores, the presence of two types of pores (small and large) over the entire cuticle in the new species vs small pores present only in the anterior part of the body and large pores present only in the posterior part of the body in M. harrylewisi, the presence of a single large pore on each of legs I—HI, stylet supports inserted in Tardigrades from the Baltic States 97 a more anterior position (pt=49.5—56.2 in the new species vs pt=61.4—67.6 in M. harrylewisi), a different macroplacoid length sequence (2<1<3 in the new species vs 2<3<1 um in M. harrylewisi), a different shape of egg processes (short, single- tipped cones in the new species vs elongated, tapering cones with bulbous bases in M. harrylewisi), a smaller diameter of eggs without and with processes (44.1—45.7 um and 55.1—55.6 um in the new species vs 66.1—80.0 um and 78.2—101.9 um in M. harrylewisi), a slightly lower number of processes on egg circumference (29-30 in the new species vs 32-41 in M. harrylewisi), and by smaller egg processes (4.5— 5.3 um in the new species vs 7.6-12.8 um in M. harrylewisi). M. jonesorum Meyer et al., 2011 by: the absence of trilobed and polygonal pores, the presence of two types of cuticular pores (small and large) in the new species vs small pores present only in the anterior part of the body, intermediate in size in the middle of the body and large pores in the posterior part of the body in M. jonesorum), the presence of a single large pore on each of legs I-III, the pres- ence of granulation on all legs, a slightly shorter buccal tube (17.7—22.9 um in the new species vs 24.4—29.6 um in M. jonesorum), stylet supports inserted in a more anterior position (pt=49.5—56.2 in the new species vs pt=63.0-65.6 in M. jonesorum), a slightly smaller external diameter of the buccal tube (1.3—2.0 um [pt=7.0-9.9] in the new species vs 2.1—-2.6 um [pt=7.4-10.7] in M. jonesorum), a different macroplacoid length sequence (2<1<3 in the new species vs 1<2<3 in M. jonesorum), slightly shorter macroplacoids I and III (II: 1.2-1.7 um [pt=6.8-8.6]; III: 1.4-2.2 um [pt=7.4-9.6] in the new species vs II: 1.9-2.3 um [pt=7. 1-8. 8]; III: 2.4-2.6 um [pt=8.4-9.9] in M. jonesorum), a slightly shorter macroplacoid row (4.5—-6.8 um [24.3-33.7] in the new species vs 7.0-8.4 um [pt=27.0-34.4] in M. jonesorum), the presence of a microplacoid, and by slightly shorter primary and secondary branches of external claws I-IV (compare Table 2 below and Table 2 in Meyer et al. 2011). M. keppelensis Claxton, 1998 by: the lack of red pigment granules, the presence of two types of cuticular pores (small and large) in the new species vs pores uni- form in size (ca. 1.0 um) in M. keppelensis), the presence of a single large pore on each of legs I-II], a slightly shorter buccal tube (17.7—22.9 um in the new species vs 24.9-28.4 um in M. keppelensis), stylet supports inserted in a more anterior position (pt=49.5—56.2 in the new species vs pt=60.6 in M. keppelensis), a different macroplacoid length sequence (2<1<3 in the new species vs 2=3<1 in M. keppelen- sis), a slightly shorter macroplacoid row (4.5—6.8 um in the new species vs 7.0—7.6 um in MM. keppelensis), the lack of a membrane around egg processes, a smaller diameter of eggs with processes (55.1—55.6 um in the new species vs 65.0—-85.0 um in MM. keppelensis), a larger number of processes on egg circumference (29-30 in the new species vs ca. 11 in M. keppelensis), smaller egg processes (4.5—5.3 wm in the new species vs 11.0—-16.0 um in M. keppelensis), narrower egg processes bases (2.4—3.4 um in the new species vs 9.0-12.0 um in M. keppelensis), and by slightly smaller distances between egg processes (1.8—3.9 um in the new species vs 4.0—6.0 um in M. keppelensis). 98 Krzysztof Zawierucha et al. / ZooKeys 408: 81-105 (2014) M. orthofasciatus Fontoura et al., 2009 by: cuticular pores arranged in 9-10 trans- verse bands (11 transverse bands present in /. orthofasciatus), the absence of tri- and quadrilobed cuticular pores, the presence of two types of pores (small and large) in the new species vs all pores of similar size in M. orthofasciatus, the presence of a single large pore on each of legs I-III, the presence of granulation on all legs, stylet supports inserted in a more anterior position (pt=49. 5—56.2 in the new species vs pt=66.5—67.8 in M. orthofasciatus), a different shape of egg processes (short, single tip cones without a membrane in the new species vs screw-like processes with a membrane and six areoles in M. orthofasciatus), a slightly larger number of processes on egg circumference (29-30 in the new species vs ca. 24 in M. orthofasciatus), and by smaller distances between egg processes (1.8—3.9 um in the new species vs 6.4-6.9 um in M. orthofasciatus). M. poricinctus Claxton, 1998 by: cuticular pores arranged in 9-10 transverse bands (8 transverse bands in M. poricinctus), the presence of two types of pores (small and large) in the new species vs uniform pore size in M. poricinctus), the presence of a single large pore on each of legs I-III, stylet supports inserted in a more anterior position (pt=49.5—56.2 in the new species vs pt=59.5 in M. poricinc- tus), a different macroplacoid length sequence (2<1<3 in the new species vs 2=3<1 in M. poricinctus), a different shape of egg processes (short, single-tipped cones without a membrane in the new species vs. screw-like processes within a mem- brane in M. poricinctus), the absence of granulation on egg shell, a larger number of processes on egg circumference (29-30 in the new species vs 18—20 in M. por- icinctus), slightly smaller egg processes (4.5—5.3 um in the new species vs 6.5—7.0 um in M. poricinctus), and by smaller distances between egg processes (1.8—3.9 um in the new species vs 6.0-8.0 um in M. poricinctus). M. pustulatus (Ramazzotti, 1959) by: the absence of triangular and polygonal pores, the presence of two types of cuticular pores (small and large) in the new species vs small pores present only in the anterior part of the body, intermediate in size in the middle of the body and the large pores in the posterior part of the body in MV. pustulatus), the presence of a single large pore on each of legs I-III and, egg processes without a filiform bristle. M. ramazzottii Binda & Pilato, 1992 by: pores arranged in bands, the presence of two types of pores (small and large) in the new species vs universal pores size in M. ramazzottii), the presence of a single large pore on each of legs I-III, the absence of the oral cavity armature, stylet supports inserted in a more anterior position (pt=49.5—56.2 in the new species vs pt=68.2—-68.3 in M. ramazzottii), a different macroplacoid length sequence (2<1<3 in the new species vs 3<2<1 in M. ramazzot- tii), and by a lower number of processes on egg circumference (29-30 in the new species vs ca. 34—41 in M. ramazzottii). M. subintermedius (Ramazzotti, 1962) by the presence of cuticular pores, the presence of granulation on all legs, and by fully developed lunules (only small open lunules present in M. subintermedius). M. vinciguerrae Binda & Pilato, 1992 by: pores arranged in bands, the absence of tri- and quadrilobed pores, the presence of two types of pores (small and large) Tardigrades from the Baltic States 99 in the new species vs uniform pore size in M. vinciguerrae), the presence of a single large pore on each of legs I-IH, the absence of the oral cavity armature, a larger mean body size (184 um in the new species vs 380 um in M. vinciguerrae), stylet supports inserted in a more anterior position (pt=49.5—56.2 in the new species vs pt=66.1-68.7 in M. vinciguerrae), a different macroplacoid length sequence (2<1<3 in the new species vs 2<3<1 um in M. vinciguerrae), a smaller diameter of eggs without and with processes (44.1—45.7 um and 55.1—55.6 um in the new species vs ca. 76.4 um and ca. 88.0 um in M. vinciguerrae), a slightly larger number of processes on egg circumference (29-30 in the new species vs ca. 26 in M. vin- ciguerrae), egg processes without flexible filaments, smaller egg processes (4.5—5.3 um in the new species vs ca. 8.2 um in M. vinciguerrae), and by narrower bases of ege processes (2.4—3.4 um in the new species vs ca. 5.0 um in M. vinciguerrae). M. weglarskae Michalczyk et al., 2005 by: the absence of bi-, trilobed and star- shaped pores, the presence of two types of pores (small and large) in the new species vs uniform pore size in MM. weglarskae), the absence of 3—5 large triangular or irregu- larly shaped pores on the caudo-dorsal cuticle above hind legs, the presence of a sin- gle large pore on each of legs I-III, a different shape of egg processes (short, single tip cones without a membrane in the new species vs. screw-like processes within a mem- brane in M. weglarskae), a slightly larger number of processes on egg circumference (29-30 in the new species vs ca. 24 in M.. weglarskae), and by slightly wider bases of ege processes (2.4—3.4 tum in the new species vs 1.6—2.0 um in M. weglarskae). M. xavieri Fontoura et al., 2009 by: the absence of trilobed pores, the presence of two types of pores (small and large) in the new species vs all pores of similar size in M. xavieri), the presence of a single large pore on each of legs I-III, the presence of granulation on all legs, a smaller body size (113-236 um in the new species vs 275-410 um in M. xavieri), stylet supports inserted in a more anterior posi- tion (pt=49. 5—56.2 in the new species vs pt=66.1-67.9 in M. xavieri), a different macroplacoid length sequence (2<1<3 in the new species vs 2<3<1 in M. xavieri). shorter macroplacoids (I: 1.3-1.9 um [pt=6.9-8. 3]; II: 1.2-1.7 pm [pt=6.8-8.6] III: 1.4-2.2 um [pt=7.4-9.6] in the new species vs I: 3.6-4.5 wm [12.7—13.8]; I: 2.9-3.6 um [10.3—11.1] III: 3.0-3.9 um [pt=10.9-11.9] in M. xavieri), a shorter microplacoid (0.5—0.9 um [pt=2.4-4.7] in the new species vs 1.5—2.0 [5.0-6.2] in M. xavieri), a shorter macroplacoid row (4.5-6.8 um [pt=24.3-33.7] in the new species vs 9.8-12.6 um [pt=35.6—38.5] in M. xavieri), a shorter placoid row (5.2-7.9 um [pt=27. 1-39.1] in the new species vs 10.9-13.9 um [39.6—-43.3] in M. xavieri), a different shape of egg processes (short, single-tipped cones in the new species vs long cones with bi- or multi-tipped tips in M. xavieri), egg shell and processes without granulation, a smaller diameter of eggs without and with pro- cesses (44.1—45.7 um and 55.1—55.6 um in the new species vs 56.0—79.0 um and 80.0-99.2 um in M. xavieri), a larger number of processes on egg circumference (29-30 in the new species vs 20-23 in M. xavieri), smaller egg processes (4.5—5.3 uum in the new species vs 10.6—-19.0 um in M. xavieri), and by slightly narrower bases of egg processes (2.4—3.4 um in the new species vs 3.7—6.6 um in M. xavieri). 100 Krzysztof Zawierucha et al. / ZooKeys 408: 81-105 (2014) Genus: Paramacrobiotus Guidetti, Schill, Bertolani, Dandekar & Wolf, 2009 Paramacrobiotus richtersi (Murray, 1911) Localities and specimen numbers. XI: 2 specimens (including 1 simplex) and 1 egg. Remarks. Paramacrobiotus species (until recently a collection of species within Macrobiotus) can be divided into three groups: areolatus, huziori and richtersi, with re- spect to the combination of two traits: the presence/absence of the microplacoid in the pharynx and the type of egg areolation. Paramacrobiotus richtersi, considered cosmo- politan, is recognised as the nominal species for a group of very similar taxa that require careful taxonomic examination of adults and egg morphology for correct identification. In the last decade many new species of this group have been described from various localities (e.g. Pilato et al. 2004, Kaczmarek et al. 2005, Tumanov 2005b, Michalczyk and Kaczmarek 2006a, b, Michalczyk et al. 2006, Pilato et al. 2006a, b, Degma et al. 2008, Bartels et al. 2009, Pilato et al. 2012), with more recent additions including molecular data (Guidetti et al. 2009, Schill et al. 2010). Acknowledgements KZ would like to express sincere thanks to Malgorzata Kuznik for her help in preparing figures. References Argue CW (1974) Tardigrades from New Brunswick, Canada. 3. Canadian Journal of Zoology 52: 919-992. doi: 10.1139/z74-122 Bartels PJ, Pilato G, Lisi O, Nelson DR (2009) Macrobiotus (Eutardigrada, Macrobiotidae) from the Great Smoky Mountains National Park, Tennessee/North Carolina, USA (North America): two new species and six new records. Zootaxa 202: 45-57. Beasley CW, Kaczmarek L, Michalczyk £ (2008) Doryphoribius mexicanus, a new species of Tardigrada (Eutardigrada: Hypsibiidae) from Mexico (North America). Proceedings of the Biological Society of Washington 121(1): 34-40. doi: 10.2988/07-30.1 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. doi: 10.1111/j.1463-6409.1993.tb00347.x Bertolani R, Rebecchi L, Giovannini I, Cesari M (2011) DNA barcoding and integrative tax- onomy 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 (1992) Minibiotus furcatus, nuova posizione sistematica per Macrobiotus furcatus Ehrenberg, 1859, e descrizione di due nuove specie. Animalia 19: 111-120. Tardigrades from the Baltic States 101 Biserov VI, Pilato G, Lisi O (2011) Macrobiotus turnovae sp. n., a new species of tardigrade from Russia. Invertebrate Zoology 8(1): 57-62. Cuénot L (1929) Description dun tardigrade nouveau de la faune francaise. Archives d’anatomie Microscopique 25: 121-125. Cuénot L (1932) Tardigrades. In: Lechevalier P (Ed) Faune de France 24: 1-96. Claxton SK (1998) A revision of the genus Minibiotus (Tardigrada: Macrobiotidae) with descrip- tions of eleven new species from Australia. Records of the Australian Museum 50: 125-160. doi: 10.3853/j.0067-1975.50.1998.1276 Dastych H (1980) Niesporczaki (Tardigrada) Tatrzaniskiego Parku Narodowego. Monografie Fauny Polski 9: 1-232. Dastych H (1988) The Tardigrada of Poland. Monografie Fauny Polski 16: 1-255. Dastych H (1990) Lsohypsibius sattleri (Richters 1902), a valid species (Tardigrada). Sencken- bergiana Biologica 71: 181-189. Dastych H (2002) A new species of the genus Macrobiotus Schultze, 1834 from Iles Kerguélen, in the sub-Antarctic (Tardigrada). Mitteilungen aus dem Hamburgischen Zoologischen Museum und Institut 99: 11—27. Dastych H (2005) Macrobiotus ramoli sp. n., a new tardigrade species from the nival zone of the Otztal Alps, Austria (Tardigrada). Mitteilungen aus dem Hamburgischen Zoologischen Museum und Institut 102: 21-35. Degma P, Bertolani R, Guidetti R (2013) Actual checklist of Tardigrada species. Ver. 23: 15-07-2013. http://www.tardigrada.modena.unimo.it/miscellanea/Actual%20check- list% 200f%20Tardigrada.pdf Degma P, Michalczyk £, Kaczmarek L (2008) Macrobiotus derkai, a new species of Tardigrada (Eutardigrada, Macrobiotidae, /uzori group) from the Colombian Andes (South America). Zootaxa 1731: 1-23. Ehrenberg CG (1859) Beitrag zur Bestimmung des stationdren mikroscopischen Lebens in bis 20,000 Fuss Alpenhéhe. Abhandlungen der K6niglichen Akademie der Wissenschaften in Berlin 1858: 429-456. Fontoura P, Pilato G (2007) Diphascon (Diphascon) faialense sp. n. a new species of Tardigrada (Eutardigrada, Hypsibiidae) from Azores and a key to the species of the D. pingue group. Zootaxa 1589: 47-55. Fontoura P, Pilato G, Lisi O (2008) New records of eutardigrades (Tardigrada) from Faial and Pico Islands, the Azores, with the description of two new species. Zootaxa 1778: 37-47. Fontoura P, Pilato G, Lisi O, Morais P (2009a) Tardigrades from Portugal: four new records and description of two new species. Zootaxa 2030: 21-38. Fontoura P, Pilato G, Morais P, Lisi O (2009b) Minibiotus xavieri, a new species of tardigrade from Parque Bioldgico de Gaia, Portugal (Eutardigrada: Macrobiotidae). Zootaxa 2267: 55-64. Guidetti R, Bertolani R, Degma P (2007) New taxonomic position of several Macrobiotus spe- cies (Eutardigrada: Macrobiotidae). Zootaxa 1471: 61-68. Guidetti R, Schill RO, Bertolani R, Dandekar T, Wolf M (2009) New molecular data for tardi- grade phylogeny, with the erection of Paramacrobiotus gen. nov. Journal of Zoological Sys- tematics and Evolutionary Research 47: 315-321. doi: 10.1111/j.1439-0469.2009.00526.x 102 Krzysztof Zawierucha et al. / ZooKeys 408: 81-105 (2014) 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: Macrobioti- dae). Journal of Natural History. doi: 10.1080/00222933.2013.800610 Guil N, Guidetti R (2005) A new species of Tardigrada (Eutardigrada: Macrobiotidae) from Iberian Peninsula and Canary Islands (Spain). Zootaxa 889: 1-11. Holt BG, Lessard J-P, Borregaard KM, Fritz SA, Aratjo MB, Dimitrov D, Fabre P-H, Graham CH, Graves GR, Jonsson KA, Nogués-Bravo D, Wang Z, Whittaker RJ, Fjeldsa J, Rah- bek C (2012) An Update of Wallace’s Zoogeographic Regions of the World. Science 339 (6115): 74-78. doi: 10.1126/science. 1228282 Horning DS, Schuster RO, Grigarick AA (1978) Tardigrada of New Zealand. New Zealand Journal of Zoology 5: 185—280. doi: 10.1080/03014223.1978.10428316 Kaczmarek L, Michalczyk L (2004) New records of Tardigrada from Cyprus with a description of the new species Macrobiotus marlenae (hufelandi group) (Eutardigrada: Macrobiotidae). Genus 15(1): 141-152. Kaczmarek L, Michalczyk L (2009a) Redescription of Hypsibius microps Thulin, 1928 and H. pallidus Thulin, 1911 (Eutardigrada: Hypsibiidae) based on the type material from the Thulin collection. Zootaxa 2275: 60-68. Kaczmarek L, Michalczyk L (2009b) Two new species of Macrobiotidae, Macrobiotus szeptyckii (harmsworthi group) and Macrobiotus kazmierskii (hufelandi group) from Argentina. Acta Zoologica Cracoviensia 52B: 87-99. doi: 10.3409/azc.52b_1-2.87-99 Kaczmarek L, Michalczyk £, Degma P (2007) Description of a new tardigrade, Macrobiotus barbarae (Eutardigrada: Macrobiotidae), from the Dominican Republic. Annales Zoologici 57(3): 363-369. Kaczmarek L, Michalczyk L, Diduszko D (2005) Some tardigrades from Siberia (Russia, Baikal region) with a description of Macrobiotus garynahi sp. n. (Eutardigrada: Macrobiotidae: richtersi group). Zootaxa 1053: 35-45. Kaczmarek L, Michalczyk L, McInnes SJ (2014a) Annotated zoogeography of non-marine Tar- digrada. Part I: Central America. Zootaxa 3763(1): 1-62. doi: 10.11646/zootaxa.3763.1.1 Kaczmarek L, Cytan J, Zawierucha K, Diduszko D, Michalczyk L (2014b) Tardigrades from Peru (South America), with descriptions of three new species of Parachela. Zootaxa 3790 (2): 357-379. doi: 10.11646/zootaxa.3790.2.5 Kaczmarek L, Goldyn B, Prokop ZM, Michalczyk £ (2011) New records of Tardigrada from Bulgaria with the description of Macrobiotus binieki sp. n. (Eutardigrada: Macrobiotidae) and a key to the species of the harmsworthi group. Zootaxa 2781: 29-39. Kaczmarek L, Zawierucha K, Smykla J, Michalczyk £ (2012) Tardigrada of the Revdalen (Spitsbergen) with the descriptions of two new species: Bryodelphax parvuspolaris (Hetero- tardigrada) and /sohypsibius coulsoni (Eutardigrada). Polar Biology 35(7): 1013-1026. doi: 10.1007/s00300-01 1-1149-0 Li X, Wang D, Wang L (2008) The Tardigrada fauna of Hainan Island (Asia: China) with descriptions of two new species. Raffles Bulletin of Zoology 56(2): 293-305. Marcus E (1930) Beitrage zur Tardigradensystematik. Zoologische Jahrbuecher Systematik 59: 363-386. Tardigrades from the Baltic States 103 Marley NJ, McInnes SJ, Sands CJ (2011) Phylum Tardigrada: A re-evaluation of the Parachela. Zootaxa 2819: 51-64. McInnes SJ (1994) Zoogeographic distribution of terrestrial/freshwater tardigrades from current literature. Journal of Natural History 28: 257-352. doi: 10.1080/00222939400770131 Meyer HA (2012) A new water bear, Minibiotus julianae, from the Caribbean Island of Domi- nica (Tardigrada: Eutardigrada: Parachela: Macrobiotidae). Proceedings of the Biological Society of Washington 125(1): 54-60. doi: 10.2988/11-15.1 Meyer HA, Domingue MD (2011) Minibiotus acadianus (Eutardigrada: Macrobiotidae), a new species of Tardigrada from southern Louisiana, USA. Western North American Naturalist 71(1): 38-43. doi: 10.3398/064.07 1.0106 Meyer HA, Hinton JG (2009) The Tardigrada of southern Africa, with the description of Minibiotus harrylewisi, a new species from KwaZulu-Natal, South Africa (Eutardigrada: Macrobiotidae). African Invertebrates 50(2): 255-268. doi: 10.5733/afin.050.0203 Meyer HA, Lyons AM, Nelson DR, Hinton JG (2011) Tardigrada of Michigan, Northern USA, with the description of Minibiotus jonesorum sp. n. (Eutardigrada: Macrobiotidae). Journal of Zoological Systematics and Evolutionary Research 49(S1): 40-47. doi: 10.1111/j.1439- 0469.2010.00596.x Michalezyk L, Kaczmarek L (2003a) Minibiotus constellatus, a new species of Tardigrada (Eu- tardigrada, Macrobiotidae) from Peru. Genus 14(2): 295-305. Michalczyk £, Kaczmarek £ (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. Michalezyk L, Kaczmarek £ (2004) Minibiotus eichhorni sp. n., a new species of eutardigrade (Eutardigrada: Macrobiotidae) from Peru. Annales Zoologici 54: 673-676. Michalczyk £, Kaczmarek L (2006a) Macrobiotus huziori, a new species of Tardigrada (Eutardi- grada: Macrobiotidae) from Costa Rica (Central America). Zootaxa 1169: 47-59. Michalczyk L, Kaczmarek L (2006b) A new species Macrobiotus magdalenae (Tardigrada: Eu- tardigrada: Macrobiotidae, richtersi group) from Costa Rican rain forest (Central America). New Zealand Journal of Zoology 33: 189-196. doi: 10.1080/03014223.2006.9518444 Michalezyk L, Kaczmarek L (2013) The Tardigrada Register: a comprehensive online data re- pository for tardigrade taxonomy. Journal of Limnology 72(S1): 175-181. doi: 10.4081/ jlimnol.2013.s1.e22 Michalczyk £, Kaczmarek £, Claxton SK (2005) Minibiotus weglarskae, a new species of Tardigrada (Eutardigrada: Macrobiotidae) from Mongolia. Zootaxa 1008: 47-56. Michalczyk £, Kaczmarek L, Weglarskae B (2006) Macrobiotus sklodowskae sp. n. (Tardigrada: Eutardigrada: Macrobiotidae, richtersi group) from Cyprus. Zootaxa 1371: 45—46. Michalczyk L, Wetnicz W, Frohme M, Kaczmarek £ (2012a) Redescriptions of three Milnesium Doyéere, 1840 taxa (Tardigrada: Eutardigrada: Milnesiidae), including the nominal species for the genus. Zootaxa 3154: 1-20. Michalczyk L, Welnicz W, Frohme M, Kaczmarek £ (2012b) Corrigenda of Zootaxa 3154: 1-20 Redescriptions of three Milnesium Doyére, 1840 taxa (Tardigrada: Eutardigrada: Milnesiidae), including the nominal species for the genus. Zootaxa 3393: 66-68. 104 Krzysztof Zawierucha et al. / ZooKeys 408: 81-105 (2014) Miller WR, McInnes SJ, Bergstrom, DM (2005) Tardigrades of the Australian Antarctic: Hypsibius heardensis (Eutardigrada: Hypsibiidae: dujardini group) a new species from sub- Antarctic Heard Island. Zootaxa 1022: 57-64. Murray J (1905) The Tardigrada of the Forth Valley. The Annals of Scottish Natural History 55: 160-164. Pilato G (1981) Analisi di nuovi caratteri nello studio degli Eutardigradi. Animalia 8: 51-57. Pilato G (1987) Revision of the genus Diphascon Plate, 1889, with remarks on the subfamily Itaquasconinae (Eutardigrada, Hypsibiidae). In: Bertolani (Ed) Biology of Tardigrades. Selected Symposia and Monographs U.Z.L., 1: 337-357. Pilato G (2013) The taxonomic value of the structures for the insertion of the stylet muscles in the Eutardigrada, and description of a new genus. Zootaxa 3721(4): 365-378. doi: 10.11646/zootaxa.3721.4.4 Pilato G, Binda MG (1997/1998) A comparison of Diphascon (D.) alpinum Murray, 1906, D. (D.) chilenense Plate, 1889 and D. (D.) pingue Marcus, 1936 (Tardigrada), and description of a new species. Zoologischer Anzeiger 236: 181-185. Pilato G, Binda MG (1999) Three new species of Diphascon of the pingue group (Eutardigrada, Hypsibiidae) from Antarctica. Polar Biology 21: 335-342. doi: 10.1007/s003000050370 Pilato G, Claxton SK (1988) Tardigrades from Australia. 1. Macrobiotus hieronimi and Min- ibiotus maculartus, two new species of eutardigrades. Animalia 15: 83-89. Pilato G, Lisi O (2006a) Macrobiotus rigidus sp. n., a new species of eutardigrade from New Zealand. Zootaxa 1109: 49-55. Pilato G, Lisi O (2006b) Notes on some tardigrades from southern Mexico with description of three new species. Zootaxa 1236: 53-68. Pilato G, Lisi O (2009a) Tardigrades of the Seychelles Islands, with the description of three new species. Zootaxa 2124: 1-20. Pilato G, Lisi O (2009b) Description of three new species of Tardigrada from Seychelles. Zootaxa 2005: 24—34. Pilato G, Binda MG, Lisi O (2003) Remarks on some species of tardigrades from South America with the description of Minibiotus sidereus n. sp. Zootaxa 195: 1-8. Pilato G, Binda MG, Lisi O (2004) Notes on tardigrades of the Seychelles with the description of three new species. Italian Journal of Zoology 71: 171-178. doi: 10.1080/11250000409356569 Pilato G, Binda MG, Lisi O (2006a) Three new species of eutardigrades from the Seychelles. New Zealand Journal of Zoology 33: 39-48. doi: 10.1080/03014223.2006.9518429 Pilato G, Binda MG, Lisi O (2006b) Eutardigrada from New Zealand, with descriptions of two new species. New Zealand Journal of Zoology 33: 49-63. doi: 10.1080/03014223.2006.95 18430 Pilato G, Binda MG, Napolitano A, Moncada E (2000) The specific value of Macrobiotus coronatus DeBarros 1942, and description of two new species of the harmsworthi group (Eutardigrada). Bollettino delle sedute della Accademia Gioenia di Scienze Naturali in Catania 33: 103-120. Pilato G, Kaczmarek L, Michalczyk L, Lisi O (2003) Macrobiotus polonicus, a new species of Tardi- grada 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. Tardigrades from the Baltic States 105 Ramazzotti G (1959) Il gruppo dell’ Echiniscus viridis con la nuova species E. perviridis e Macro- biotus pustulatus altra nuova specie (Tardigrada). Atti della Societa Italiana di Scienze Natu- rali e del Museo Civico di Storia Naturale in Milano 98: 303-309. Ramazzotti G (1962) Tardigradi del Cile, con descrizione di quattro nuove specie e di una va- rieta. Atti della Societa Italiana di Scienze Naturali e del Museo Civico di Storia Naturale di Milano 101: 275-287. Ramazzotti G, Maucci W (1983) I Phylum Tardigrada. Memorie dell’Istituto Italiano di Idrobiologia 41: 1-1012. Rossi G, Claps M, Ardohain D (2009) Tardigrades from northwestern Patagonia (Neuquén Province, Argentina) with the description of three new species. Zootaxa 2095: 21-36. Satkauskiené I, Vosylitité R (2010) Microfauna of Moss (Bryophyta: Bryopsida) from four regions of Lithuania. Acta Zoologica Lituanica 20: 225-231. doi: 10.2478/v10043-010- 0024-5 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 Paramac- robiotus (Tardigrada). Organisms Diversity & Evolution 10(4): 287-296. doi: 10.1007/ s13127-010-0025-z Tumanov DV (2005a) Two new species of Macrobiotus (Eutardigrada, Macrobiotidae) from Tien Shan (Kirghizia), with notes on Macrobiotus tenuis group. Zootaxa 1043: 33-46. Tumanov DV (2005b) Notes on the Tardigrada of Thailand, with description of Macrobiotus alekseevi sp. n. (Eutardigrada, Macrobiotidae). Zootaxa 999: 1-16. Tumanov DV (2006) Five new species of the genus Milnesium (Tardigrada, Eutardigrada, Milnesiidae). Zootaxa 1122: 1-23. Zawierucha K, Kazmierski A (2012) ‘The first records of tardigrades (Tardigrada, Eutardigrada) from Estonia. Zoology and Ecology 22: 111-113. doi: 10.1080/21658005.2012.696885 Ziemelis A, Purina L, Ozoling A (2012) A Short-term study of population Dynamics of tar- digrades in the moss Leucodon sciuroides (Hedw.) SCHWAGR. Latvijas Universitates 70. zinatniska konference Biolodijas sekcija, Zooloaijas un dzivnieku ekoloaijas apakssekcija (conference abstract).