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A PDF file describing our methods in greater detail, alongside the new opsin family nomenclature

A single PDF file containing Supplemental Figures 1 through 8, as described in the main text, alongside their figure legends

The zappalai group: 31(3). Dorsal cuticle smooth or slightly rugose.................................................................. 32 -. Dorsal cuticle with small tubercles............................................................... D. taiwanus 32(31). Oral cavity armature present............................................................................33 -. Oral cavity armature absent or not visible under LM........................................................ 35 33(32). Oral cavity armature composed of a single dorsal tooth.......................................................34 -. Oral cavity armature with one anterior band of large teeth and one single medio-dorsal tooth in the posterior band......... ........................................................................................ D. turkmenicus 34(33). Configuration of macroplacoids 1> 2> 3, secondary branches of claws longer than 50% of the primary branches.......... .......................................................................................... D. mexicanus -. Configuration of macroplacoids 2> 1> 3, secondary branches of claws shorter than 50% of the primary branches......... ........................................................................................... D. bertolanii 35(31). Lunules on claws present, freshwater species........................................................ D. zappalai -. Lunules on legs absent, terrestrial species................................................................. 36 36(35). Claws without enlarged bases, ratio of the secondary to the primary branches more than 0.79............... D. longistipes -. Claws with enlarged bases............................................................................ 37 37(36). Long claws with a wide bulbous base.................................................. D. chetumalensis sp. nov. -. Claws without a wide bulbous base and with a ratio of the secondary to the primary branches less than 0.70............ 38 38(37). Ratio of the secondary to the primary branches of claws more than 0.60............................... D. qinlingense -. Ratio of the secondary to the primary branches of claws less than 0.55................................. D. korganovae, Published as part of P��rez-Pech, Wilbert Andr��s, Anguas-Escalante, Abril, Cutz-Pool, Leopoldo Querubin & Guidetti, Roberto, 2017, Doryphoribius chetumalensis sp. nov. (Eutardigrada: Isohypsibiidae) a new tardigrade species discovered in an unusual habitat of urban areas of Mexico, pp. 345-356 in Zootaxa 4344 (2) on page 354, DOI: 10.11646/zootaxa.4344.2.9, http://zenodo.org/record/1043140

The doryphorus group: 17(2). Claws IV reduced and clearly smaller than claws I���III................................................. D. pilatoi -. Claws IV similar in size or larger than claws I���III.......................................................... 18 18(17). Entire cuticle without sculpture.........................................................................19 -. Cuticle at least partially sculptured...................................................................... 22 19(18). Lunules on internal claws I���III present, on external absent............................................ D. koreanus -. Internal and external claws without lunules............................................................... 20 20(19). Stylet furcae robust, triangular in shape, with the postero-lateral processes rounded but not swollen.......... D. neglectus -. Stylet furcae with swollen postero-lateral processes.........................................................21 21(20). Oral cavity armature visible, and one large dorso-median tooth at the beginning of the buccal tube present, buccal tube 3 ��m or more in width.............................................................................. D. macrodon -. Oral cavity armature absent/not visible under LM, buccal tube less than 3 ��m in width.................... D. doryphorus 22(18). Dorsal cuticle smooth but large granules present on legs IV........................................ D. tergumrudis -. Dorsal cuticle sculptured.............................................................................. 23 23(22). Ventral sculpture present, lunules on claws present, pt of buccal tube width less than 15, eyes present........ D. polynettae -. Ventral sculpture absent...............................................................................24 24(23). Lunules on claws absent, pt of buccal tube external width more than 20, eyes absent..................... D. smokiensis -. Lunules present, those of the internal claws of the first three pairs of legs often could be difficult to see............... 25 25(24). A cuticular bar at the base of the inner claws on the first three pairs of legs present, evident lunules of the inner claws...... ......................................................................................... D. solidunguis -. A cuticular bar at the base of the inner claws on the first three pairs of legs absent.................................26 26(25). Cuticle with a delicate reticulate sculpture of very thin ridges forming a mesh with a tiny tubercle at each crossing, sculptured legs........................................................................................ D. bindae -. Cuticle with an evident reticulated sculpture with mesh slightly irregular in shape and size, delimited by ridges which form a tubercle at each crossing legs not sculptured.................................................... D. amazzonicus, Published as part of P��rez-Pech, Wilbert Andr��s, Anguas-Escalante, Abril, Cutz-Pool, Leopoldo Querubin & Guidetti, Roberto, 2017, Doryphoribius chetumalensis sp. nov. (Eutardigrada: Isohypsibiidae) a new tardigrade species discovered in an unusual habitat of urban areas of Mexico, pp. 345-356 in Zootaxa 4344 (2) on pages 353-354, DOI: 10.11646/zootaxa.4344.2.9, http://zenodo.org/record/1043140

The vietnamensis group: 27(3). Eight rows of gibbosities (VIII:2-2-2-2-2-2-2-2), eyes absent........................................... D. minimus -. More than eight rows of gibbosities, eyes present........................................................... 28 28(27). Nine rows of gibbosities (IX:5-4-5-4-5-4-5-2-3), dorsal cuticle with fine granulation.................... D. vietnamensis -. More than nine rows of gibbosities, cuticle with polygonal tubercles............................................29 29(28). Ten rows of well visible gibbosities (X:4-4-4-4-4-4-4-2-2-3)............................................ D. gibber -. More than ten rows of gibbosities........................................................................30 30(29). Twelve rows of poorly visible gibbosities (XII:2-4-2-4-2-4-2-4-2-4-4-1)................................. D. mariae -. Sixteen rows of dorso-lateral gibbosities present (XVI: 3-4-2-6-2-6-2-6-2-6-6-2-6-2-3-3+2[LII-IV])......... D. elleneddiei, Published as part of P��rez-Pech, Wilbert Andr��s, Anguas-Escalante, Abril, Cutz-Pool, Leopoldo Querubin & Guidetti, Roberto, 2017, Doryphoribius chetumalensis sp. nov. (Eutardigrada: Isohypsibiidae) a new tardigrade species discovered in an unusual habitat of urban areas of Mexico, pp. 345-356 in Zootaxa 4344 (2) on page 354, DOI: 10.11646/zootaxa.4344.2.9, http://zenodo.org/record/1043140

Doryphoribius chetumalensis sp. nov. Description of holotype (Figs 2–4; Table 2). Body colour orange. Cuticle of dorsal and ventral surfaces, and legs, smooth (i.e. without gibbosities or sculpture; Fig. 2). Eye spots present (Figs 2C, 3A). Bucco-pharyngeal apparatus of Doryphoribius type (see: Pilato & Binda 2010; Guidetti et al. 2013b) (Fig. 3). Peribuccal structures and oral cavity armature absent or not visible under LM. Mouth antero-ventral. Ventral lamina present. Stylet furca of typical shape with two condyles and two proximal apophyses. Pharyngeal bulb spherical, with triangular pharyngeal apophyses, and three macroplacoids, without constrictions, and of similar length; microplacoid and septulum absent. Macroplacoids bean-shaped in lateral view; in dorsal view, the first triangular-shaped, the second rectangular-shaped, the third heart-shaped (Fig. 3). First and second macroplacoids close together, the third macroplacoid more distant. Claws long, of Isohypsibius type, similar in size and shape on all legs (Fig. 4). Claws with enlarged and wide bulbous bases; generally, the external claws of first three pairs of legs and the posterior claw of the hind legs with larger bulbous bases (Fig. 4). Claw lunules absent. Primary branches of all claws with thin, short accessory points. Morphometric data of the holotype and eight paratypes are reported in Table 2. Remarks. Paratypes, body length from 188.8 µm to 345.7 µm. The eye spots and the orange colour of the body were visible in live specimens but not in all mounted specimens. Two exuviae were collected, which had eleven and twelve smooth eggs. Type repositories. Holotype and 20 paratypes. Holotype and nine paratypes deposited in the Laboratory of Zoology, Chetumal Institute of Technology, Quintana Roo, Mexico; 11 paratypes in the Bertolani collection, Department of Life Sciences, University of Modena and Reggio Emilia, Italy. Character ind 10 ind 2 ind 5 ind 8 ind 9 ind 1 ind 6 holotype ind 4 µm pt index Type locality: Urban zone of Chetumal, Quintana Roo, Mexico, Petcacab street intersection with Tres Garantias street (site S1: 18°30'13"N, 88°18'19"W; 9 m a.s.l.). The substrate was soil sediment with a very clay texture accumulated on the street between the sidewalk and the asphalt (site S 1 in Table 1). Other sites of collection. S2, Erick Paolo Martines street intersection with Colibri street; S3, Petcacab street intersection with Polyuc street; S4, Jesus Urueta street intersection with Heriberto Frias street (see Table 1). Etymology. The name ‘ chetumalensis ’ refers to the locality in which the new species was found. Differential diagnosis. Doryphoribius chetumalensis sp. nov. differs from all the other species of the genus in the distinctly enlarged and wide bulbous base to the claws (Fig. 4). The new species belongs to the zappaloi group (see: Michalczyk & Kaczmarek 2010), having no dorsal cuticular gibbosities and three macroplacoids. With reference to the other species within this group, D. chetumalensis sp. nov. differs, not only by the wide bulbous basal claws, but also in other characters. - Doryphoribius turkmenicus Biserov, 1999 was described as “white or slightly yellow” (Biserov 1999) and with a well-developed anterior band of large teeth, while the new species has distinctly orange body colour, and has no visible oral cavity armature; - Doryphoribius mexicanus Beasley et al., 2008, and Doryphoribius bertolanii Beasley & Pilato, 1987, were described with very simple oral cavity armature, composed of a single large round dorsal tooth placed in a posterior position (Beasley et al. 2008; Beasley & Pilato 1987), which is absent in D. chetumalensis sp. nov.; - Doryphoribius zappalai Pilato, 1971 has lunules in the claws which are absent in the new species; - Doryphoribius korganovae Biserov, 1994 was described as yellowish brown with smooth or slightly rugose cuticle (Biserov 1994), while D. chetumalensis sp. nov. has an orange colour and smooth cuticle; the claw bases of D. korganovae are expanded, but not as enlarged as the new species; - Doryphoribius qinlingense Li et al., 2004 was described as colourless, and with three macroplacoids increasing in length from first to third (Li et al. 2004), in the new species the three macroplacoids are of similar size; claw bases of D. qinlingense are expanded but not enlarged and bulbous, as in the new species; - Doryphoribius longistipes Bartels et al., 2008 was described as having no lunules or cuticular thickening (Bartels et al. 2008), making the swollen and bulbous bases of D. chetumalensis sp. nov. very distinctive in comparison; - Doryphoribius taiwanus Li & Li, 2008 was described as colourless and with a cuticular sculpture of dorsal and lateral tubercles (Li & Li 2008), which are absent in D. chetumalensis sp. nov.

The evelinae group: 4(2). Cuticular gibbosities present only on legs...................................................................5 -. Cuticular gibbosities present on dorsal side of the body........................................................6 5(4). Dorsal cuticle smooth.......................................................................... D. evelinae -. Dorsal cuticle with reticular design.................................................................. D. flavus 6(4). Cuticular gibbosities indistinct and irregularly arranged.......................................................7 -. Cuticular gibbosities distinct and arranged in defined transverse rows........................................... 8 7(6). Cuticular tubercles present, lunules absent, hind claws larger than claws I���III........................ D. maranguensis -. Cuticular tubercles absent, lunules present, hind claws slightly smaller than claws I���III..................... D. picoensis 8(6). Four dorsal gibbosities arranged in two caudal rows (configuration: II:2-2)....................... D. quadrituberculatus -. More than two rows of dorsal gibbosities.................................................................. 9 9(8) Three rows of dorsal gibbosities (III:4-2-2), oral cavity with six teeth in two bands....................... D. tessellatus -. More than three rows of dorsal gibbosities located throughout the dorsal cuticle...................................10 10(9). Seven rows of gibbosities with only two gibbosities in each row (VII:2-2-2-2-2-2-2)................. D. dupliglobulatus -. More than seven rows of gibbosities...................................................................... 11 11(10). Nine rows of gibbosities...............................................................................12 -. Ten rows of gibbosities (X: 2-3-1-3-1-3-1-2-2-2).................................................. D. mcinnesae 12(11). Six gibbosities in rows I and II, a single gibbosity on each of the hind legs, (IX:6-6-4-6-4-6-4-4-2+2[LIV])..... D. dawkinsi -. Less than six gibbosities (2 or 4) in row I, no gibbosities in the hind legs.........................................13 13(12). Two gibbosities in row I; gibbosity configuration IX: 2-4-4-4-4-6-4-4-2................................. D. barbarae -. More than two gibbosities in row I..................................................................... 14 14(13). Four gibbosities in row 2; gibbosity configuration IX:4-4-4-4-4-4-4-2-2.......................... D. huangguoshuensis -. Six gibbosities in row 2................................................................................ 15 15(14). Gibbosity configuration IX: 4-6-5-6-5-6-4-2-2..................................................... D. niedbalai -. Gibbosity configuration IX: 4-6-4-6-4-6-4-4-2..............................................................16 16(15). Main branches of both external and internal claws with uniform width and the distal portion tapering gradually........... ..................................................................................... D. maasaimarensis -. Proximal portion of the main branches of both external and internal claws more wide and tapered abruptly at about half length .......................................................................................... D. zyxiglobus, Published as part of P��rez-Pech, Wilbert Andr��s, Anguas-Escalante, Abril, Cutz-Pool, Leopoldo Querubin & Guidetti, Roberto, 2017, Doryphoribius chetumalensis sp. nov. (Eutardigrada: Isohypsibiidae) a new tardigrade species discovered in an unusual habitat of urban areas of Mexico, pp. 345-356 in Zootaxa 4344 (2) on page 353, DOI: 10.11646/zootaxa.4344.2.9, http://zenodo.org/record/1043140

Doryphoribius chetumalensis sp. nov. Description of holotype (Figs 2���4; Table 2). Body colour orange. Cuticle of dorsal and ventral surfaces, and legs, smooth (i.e. without gibbosities or sculpture; Fig. 2). Eye spots present (Figs 2C, 3A). Bucco-pharyngeal apparatus of Doryphoribius type (see: Pilato & Binda 2010; Guidetti et al. 2013b) (Fig. 3). Peribuccal structures and oral cavity armature absent or not visible under LM. Mouth antero-ventral. Ventral lamina present. Stylet furca of typical shape with two condyles and two proximal apophyses. Pharyngeal bulb spherical, with triangular pharyngeal apophyses, and three macroplacoids, without constrictions, and of similar length; microplacoid and septulum absent. Macroplacoids bean-shaped in lateral view; in dorsal view, the first triangular-shaped, the second rectangular-shaped, the third heart-shaped (Fig. 3). First and second macroplacoids close together, the third macroplacoid more distant. Claws long, of Isohypsibius type, similar in size and shape on all legs (Fig. 4). Claws with enlarged and wide bulbous bases; generally, the external claws of first three pairs of legs and the posterior claw of the hind legs with larger bulbous bases (Fig. 4). Claw lunules absent. Primary branches of all claws with thin, short accessory points. Morphometric data of the holotype and eight paratypes are reported in Table 2. Remarks. Paratypes, body length from 188.8 ��m to 345.7 ��m. The eye spots and the orange colour of the body were visible in live specimens but not in all mounted specimens. Two exuviae were collected, which had eleven and twelve smooth eggs. Type repositories. Holotype and 20 paratypes. Holotype and nine paratypes deposited in the Laboratory of Zoology, Chetumal Institute of Technology, Quintana Roo, Mexico; 11 paratypes in the Bertolani collection, Department of Life Sciences, University of Modena and Reggio Emilia, Italy. Character ind 10 ind 2 ind 5 ind 8 ind 9 ind 1 ind 6 holotype ind 4 ��m pt index Type locality: Urban zone of Chetumal, Quintana Roo, Mexico, Petcacab street intersection with Tres Garantias street (site S1: 18��30'13"N, 88��18'19"W; 9 m a.s.l.). The substrate was soil sediment with a very clay texture accumulated on the street between the sidewalk and the asphalt (site S 1 in Table 1). Other sites of collection. S2, Erick Paolo Martines street intersection with Colibri street; S3, Petcacab street intersection with Polyuc street; S4, Jesus Urueta street intersection with Heriberto Frias street (see Table 1). Etymology. The name ��� chetumalensis ��� refers to the locality in which the new species was found. Differential diagnosis. Doryphoribius chetumalensis sp. nov. differs from all the other species of the genus in the distinctly enlarged and wide bulbous base to the claws (Fig. 4). The new species belongs to the zappaloi group (see: Michalczyk & Kaczmarek 2010), having no dorsal cuticular gibbosities and three macroplacoids. With reference to the other species within this group, D. chetumalensis sp. nov. differs, not only by the wide bulbous basal claws, but also in other characters. - Doryphoribius turkmenicus Biserov, 1999 was described as ���white or slightly yellow��� (Biserov 1999) and with a well-developed anterior band of large teeth, while the new species has distinctly orange body colour, and has no visible oral cavity armature; - Doryphoribius mexicanus Beasley et al., 2008, and Doryphoribius bertolanii Beasley & Pilato, 1987, were described with very simple oral cavity armature, composed of a single large round dorsal tooth placed in a posterior position (Beasley et al. 2008; Beasley & Pilato 1987), which is absent in D. chetumalensis sp. nov.; - Doryphoribius zappalai Pilato, 1971 has lunules in the claws which are absent in the new species; - Doryphoribius korganovae Biserov, 1994 was described as yellowish brown with smooth or slightly rugose cuticle (Biserov 1994), while D. chetumalensis sp. nov. has an orange colour and smooth cuticle; the claw bases of D. korganovae are expanded, but not as enlarged as the new species; - Doryphoribius qinlingense Li et al., 2004 was described as colourless, and with three macroplacoids increasing in length from first to third (Li et al. 2004), in the new species the three macroplacoids are of similar size; claw bases of D. qinlingense are expanded but not enlarged and bulbous, as in the new species; - Doryphoribius longistipes Bartels et al., 2008 was described as having no lunules or cuticular thickening (Bartels et al. 2008), making the swollen and bulbous bases of D. chetumalensis sp. nov. very distinctive in comparison; - Doryphoribius taiwanus Li & Li, 2008 was described as colourless and with a cuticular sculpture of dorsal and lateral tubercles (Li & Li 2008), which are absent in D. chetumalensis sp. nov., Published as part of P��rez-Pech, Wilbert Andr��s, Anguas-Escalante, Abril, Cutz-Pool, Leopoldo Querubin & Guidetti, Roberto, 2017, Doryphoribius chetumalensis sp. nov. (Eutardigrada: Isohypsibiidae) a new tardigrade species discovered in an unusual habitat of urban areas of Mexico, pp. 345-356 in Zootaxa 4344 (2) on pages 347-352, DOI: 10.11646/zootaxa.4344.2.9, http://zenodo.org/record/1043140, {"references":["Pilato, G. & Binda, M. G. (2010) Definition of families, subfamilies, genera and subgenera of the Eutardigrada, and keys to their identification. Zootaxa, 2404, 1 - 54.","Guidetti, R., Bertolani, R. & Rebecchi, L. (2013 b) Comparative analysis of the tardigrade feeding apparatus: adaptive convergence and evolutionary pattern of the piercing stylet system. Journal of Limnology, 72 (1), 24 - 35. https: // doi. org / 10.4081 / jlimnol. 2013. s 1. e 4","Michalczyk, L. & Kaczmarek, L. (2010) Description of Doryphoribius dawkinsi, a new species of Tardigrada (Eutardigrada: Hypsibiidae) from the Costa Rican highlands, with the key to the genus Doryphoribius. Zootaxa, 2393, 46 - 58.","Biserov, V. I. (1999) Tardigrada of Turkmenistan, with description of three new species. Zoologischer Anzeirger, 238, 157 - 167.","Beasley, C. W., Kaczmarek, L. & Michalczyk, L. (2008) Doryphoribius mexicanus, a new species of Tardigrada (Eutardigrada, Hypsibiidae) from Mexico (North America). Proceedings of the Biological Society of Washington, 121, 34 - 40. https: // doi. org / 10.2988 / 07 - 30.1","Beasley, C. W. & Pilato, G. (1987) Two new species de Doryphoribius (Eutardigrada, Hypsibiidae) from North America. Animalia, 14, 99 - 105.","Pilato, G. (1971) Su una nova specie di Doryphoribius (Eutardigrada: Hypaibiidae) e considerazioni sulla posizione filogenetica del genere. Bolletino di Zoologia, 38, 145 - 149. https: // doi. org / 10.1080 / 11250007109436964"]}

RICHTERSIIDAE FAM. NOV. Description: Double claws Y-shaped, with the two branches forming an evident common tract of variable length. Large teeth on all lunules. Buccal tube with ventral lamina and a cuticular thick on the anterior, dorsal wall of the buccal tube (which can form a large apophysis). Absence of transverse crests in the buccal armature. Two macroplacoids in the pharynx. Cuticular pores (at least in a phase of the life cycle). Eggs laid freely with cuticular processes on their surface. Type genus. Richtersius Composition. Richtersius, Diaforobiotus gen. nov., Adorybiotus (provisionally, see Remarks). Remarks. Based on molecular data, Richtersius, Diaforobiotus gen. nov., and Adorybiotus do not belong to Macrobiotidae, but the phylogenetic relationship of Adorybiotus with the other genera of the families Richtersiidae fam. nov. and Murrayidae needs to be clarified with further molecular data. Based on several morphological affinities with Richtersius and Diaforobiotus gen. nov. (i.e. a cuticular thickening on the anterior dorsal wall of the buccal tube, large teeth on all lunules, absence of transverse crests in the buccal armature, two macroplacoids in the pharynx, and presence of cuticular pores), we place Adorybiotus in this family., Published as part of Guidetti, Roberto, Rebecchi, Lorena, Bertolani, Roberto, Jonsson, Kjell Ingemar, Kristensen, Reinhardt M��bjerg & Cesari, Michele, 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, pp. 834-845 in Zoological Journal of the Linnean Society 178 (4) on page 843, DOI: 10.1111/zoj.12428, http://zenodo.org/record/5369103

DIAFOROBIOTUS GEN. NOV. Description: Peribuccal lamellae (ten) and ventral lamina present. A dorsal thickening present in the anterior portion of the buccal tube, in conjunction with a large tooth on the internal surface of the tube. Some strong, scattered round teeth present posterior to the second band of teeth of the buccal armature present. Type species. Macrobiotus islandicus Richters, 1904. Etymology. Diaforobiotus, from diaforos (Greek) = different, different from all other macrobiotoid genera. Composition. Diaforobiotus islandicus (Richters, 1904)., Published as part of Guidetti, Roberto, Rebecchi, Lorena, Bertolani, Roberto, Jonsson, Kjell Ingemar, Kristensen, Reinhardt M��bjerg & Cesari, Michele, 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, pp. 834-845 in Zoological Journal of the Linnean Society 178 (4) on page 843, DOI: 10.1111/zoj.12428, http://zenodo.org/record/5369103

DIAFOROBIOTUS GEN. NOV. Description: Peribuccal lamellae (ten) and ventral lamina present. A dorsal thickening present in the anterior portion of the buccal tube, in conjunction with a large tooth on the internal surface of the tube. Some strong, scattered round teeth present posterior to the second band of teeth of the buccal armature present. Type species. Macrobiotus islandicus Richters, 1904. Etymology. Diaforobiotus, from diaforos (Greek) = different, different from all other macrobiotoid genera. Composition. Diaforobiotus islandicus (Richters, 1904).

RICHTERSIIDAE FAM. NOV. Description: Double claws Y-shaped, with the two branches forming an evident common tract of variable length. Large teeth on all lunules. Buccal tube with ventral lamina and a cuticular thick on the anterior, dorsal wall of the buccal tube (which can form a large apophysis). Absence of transverse crests in the buccal armature. Two macroplacoids in the pharynx. Cuticular pores (at least in a phase of the life cycle). Eggs laid freely with cuticular processes on their surface. Type genus. Richtersius Composition. Richtersius, Diaforobiotus gen. nov., Adorybiotus (provisionally, see Remarks). Remarks. Based on molecular data, Richtersius, Diaforobiotus gen. nov., and Adorybiotus do not belong to Macrobiotidae, but the phylogenetic relationship of Adorybiotus with the other genera of the families Richtersiidae fam. nov. and Murrayidae needs to be clarified with further molecular data. Based on several morphological affinities with Richtersius and Diaforobiotus gen. nov. (i.e. a cuticular thickening on the anterior dorsal wall of the buccal tube, large teeth on all lunules, absence of transverse crests in the buccal armature, two macroplacoids in the pharynx, and presence of cuticular pores), we place Adorybiotus in this family.

Tardigrades occur worldwide and in a variety of ecosystems and habitats representing an important component of the micrometazoan biodiversity. Several studies documenting the occurrence of tardigrades in Sweden have been published since the first reports in early 1900, but no comprehensive summary of these studies have been published. We compiled the available information on recorded tardigrades from Sweden, using material from published studies and museum and university collections. In total, our review document 101 species of tardigrades that have been recorded from Sweden (an updated checklist of tardigrades from Sweden will be available online), of which 14 species are new records for the country. The highest number of species was recorded in the northernmost province of Lappland and the more southern provinces of Uppland and Skåne, while much lower species numbers are reported from the middle part of Sweden. This pattern probably represents biased sampling activities of biologists rather than real differences in biodiversity of tardigrades. In view of the few studies that have been made on tardigrade biodiversity in Sweden, the relatively high number of tardigrade species recorded, representing almost a tenth of the species recorded worldwide, indicates that many more species remain to be found. In this respect, more studies of the marine ecosystems along the Swedish west coast and the long Baltic Sea coastline would be of particular interest.

Nuclear magnetic resonance (NMR) spectroscopy is, in principle, a promising candidate to study the intracellular chemistry of single microscopic living entities. However, due to sensitivity limitations, NMR experiments were reported only on very few and relatively large single cells down to a minimum volume of 10 nl. Here we show NMR spectroscopy of single ova at volume scales (0.1 and 0.5 nl) where life development begins for a broad variety of animals, humans included. We demonstrate that the sensitivity achieved by miniaturized inductive NMR probes (few pmol of 1H nuclei in some hours at 7 T) is sufficient to observe chemical heterogeneities among subnanoliter ova of tardigrades. Such sensitivities should allow to non-invasively monitor variations of concentrated intracellular compounds, such as glutathione, in single mammalian zygotes., Main text: 5 pages, 4 figures. Supporting information: 3 pages, 6 figures

Thulinius cf. saltursus Sampling localities: Five specimens from sediment and periphyton in Caldwell Fork, North Carolina (813 m asl, N 35 �� 37.5813, W 83 �� 5.3610) and four specimens in sediment from Cataloochee Creek, North Carolina (788 m asl, N 35 �� 38.2405, W 83 �� 4.9303). The specimens were compared with the type material of both Thulinius saltursus (Schuster, Toftner & Grigarick, 1978) (a species previously attributed to Isohypsibius in Schuster et al. 1978) and Thulinius ruffoi (Bertolani, 1981) and look similar to both species. The buccal armature is typical of the genus and similarly the macroplacoids are rod-shaped and there are no microplacoids. The basal portions on all claws are normally developed and slender. A lunule appears to be present only at the base of the external (posterior) claw (as in T. saltursus, whereas T. ruffoi has lunules on all claws), but it is not always possible to determine the presence or absence of a lunule. In any case, currently this is the only character that distinguishes the two species and for this reason we tentatively attribute our specimens from North Carolina to T. cf. saltursus., Published as part of Bertolani, Roberto, Bartels, Paul J., Guidetti, Roberto, Cesari, Michele & Nelson, Diane R., 2014, Aquatic tardigrades in the Great Smoky Mountains National Park, North Carolina and Tennessee, U. S. A., with the description of a new species of Thulinius (Tardigrada, Isohypsibiidae), pp. 524-536 in Zootaxa 3764 (5) on pages 527-528, DOI: 10.11646/zootaxa.3764.5.2, http://zenodo.org/record/230310, {"references":["Schuster, R. O., Toftner, E. C. & Grigarick, A. A. (1978) Tardigrada of Pope Beach, Lake Tahoe, California. The Wasmann Journal of Biology, 35, 115 - 136."]}

Thulinius augusti (Murray, 1907) Sampling localities. Palmer Creek, Caldwell Fork, Rough Fork, and Cataloochee Creek in North Carolina, and Crying Creek in Tennessee. Predominantly in sediment but rarely in periphyton, this is one of the most abundant and widespread aquatic species in our collection, second only to H. cf. dujardini. Remarks. The specimens collected look very similar to the type material. In particular, the peculiar crown of marked sub-lobes that surrounds the mouth is evident (Fig. 2 A). Another similarity is the long, thin basal tract of the claws, in particular the basal tract of the external claw (Figs 2 C, 2 D). In the type material the absence of lunules is cited, whereas sometimes lunules are visible in the American specimens (Fig. 3 D). A transverse bar exists under the claws of the first three pairs of legs, which is not in contact with the claws, but at least in some specimens it is clearly separated into two parts (Figs 2 C, 3 C), which is different from the type material that has only a single bar. The other characters (posterior band of teeth followed by a single row of large teeth, buccal tube relatively wide, long pharyngeal apophyses and rod-shaped macroplacoids without microplacoid) are common to all the species of the genus known to date. Thulinius augusti has been cited several times as Macrobiotus augusti, Hypsibius augusti, Isohypsibius augusti and Pseudobiotus augusti, and has been considered cosmopolitan (McInnes 1994), but in most cases these were misidentifications as documented by Bertolani et al. (1999). Therefore, the only definite citations of the species are the type locality (Scotland) and those here in the GSMNP. Apart from the presence of thin lunules present only in some fresh American specimens, the morphologies of the Scottish and American material correspond. We want to emphasize the difficulty in identifying these structures when they are thin and buried in the leg. We believe that molecular comparisons would be definitive. We did successfully sequence a portion of the 18 S rDNA gene for three GSMNP specimens (slides C 3180 V 1, V 2, V 3; Table 3; voucher images in Fig. 4) from Cataloochee Creek (825 m asl, N 35 �� 37.881; 83 �� W 05.294), GenBank accession numbers KF 360230 - 2, and a portion of cox 1 gene for one specimen of them V 3 (KF 360229; Table 4), but we were unable to find fresh material in the type locality for comparison. This molecular information would be very useful for establishing whether T. augusti has a widespread geographic distribution or is composed of cryptic species with more limited distribution. Our data allowed us to compare the molecular analysis of three species of Thulinius: T. augusti (found in this GSMNP study), T. stephaniae (found in two locations in Sinai, Egypt) and Thulinius sp. (found in an unidentified location) (Tables 3, 4). Regarding the 18 S rDNA sequences, T. augusti clearly differs from T. stephaniae by a K 2 P distance of 2.7 ���3.0% (p-distance: 2.7���2.9 %) (Table 3). A specimen of T. augusti showed a cox 1 haplotype that differs from that of T. stephaniae by a K 2 P distance of 31.9 % (p-distance: 25.9 %) and from that of Thulinius sp. by a K 2 P distance of 28.2 % (p-distance: 23.4 %). T. stephaniae and Thulinius sp. differ by a K 2 P distance 28.0% (pdistance: 23.2 %) (Table 4). These results for cox 1 distance values are in line with the values that separate different tardigrade species (Cesari et al. 2009; Bertolani et al. 2011; Cesari et al. 2011). Type locality. Stream sediment in Road Prong, Tennessee (145 m asl, N 35 �� 36.5113, W 83 �� 27.5637). One paratype was found in sediments from the same site at Road Prong, and another was found at Kephart Prong, North Carolina (1075 m asl, N 35 �� 36.6226, W 83 �� 22.0946). Type material. Holotype (slide number 23 Mar08- 1-4) and paratypes (slide numbers 23 Mar08- 1 -3, 9 May08- 11-16) are in the collection of Roberto Bertolani at the Department of Life Sciences, University of Modena and Reggio Emilia. Coll. Margaret Phillips, 23 -Mar-08 and 9 -May-08. Diagnosis. Large body size (328.7���542.3 ��m in length), body shape sometimes suggesting a fat piglet (Fig. 4 A). Cuticle sculptured on all surfaces (dorsally, ventrally and on all legs) with polygonal mesh (Fig. 4 C) forming an extended roughness where the cuticle is contracted. Eyes not observed in the mounted material. Peribuccal lamellae present. Lobes around the mouth not subdivided. Buccal armature with only the posterior band of small teeth followed by a line of large rounded teeth (Fig. 4 B). Large straight buccal tube. Large pharyngeal apophyses developed longitudinally; three long rod-shaped macroplacoids; no microplacoid. The curvatures of the placoid rows resemble a Grecian urn (Fig. 4 B). Small stylet furcae (Fig. 4 B). Large claws of Isohypsibius type (Figs 5 A��� C), the external claw more slender than the internal. Primary branch of the internal claw with a distinct dorsal knob. Accessory points on the primary branches present. Basal portions of all claws slender, wider in their distal portion in the internal claw (Figs 5 A���B). Lunules probably absent on all legs (Figs 5 A���C), or present only on the external or posterior claw but very reduced. Long cuticular bars, subdivided into two parts, present under the claws on the first three pairs of legs (Figs 5 A���B). Eggs unknown. Description of the holotype: (for measurements of the sclerified structures see Table 2). Adult 357.6 ��m in length. Cuticle sculptured, with polygonal mesh dorsally, ventrally and on all legs. Eyes not observed in the mounted specimen. Peribuccal lamellae present but the number was not detectable. Buccal armature composed of only the posterior band of small teeth, followed by a line of large rounded teeth (see also taxonomic remarks) (Fig. 4 B). Buccal tube with thin walls and with long stylet supports inserted at 62.9 % of its length. Pharynx apophyses 5.5 ��m in length. Three macroplacoids present, first is the longest, the second is the shortest. The distance between second and third macroplacoid larger than between the first and second macroplacoid. Microplacoid absent. Double-claws of Isohypsibius type. Primary branch of the internal claw with a distinct dorsal knob; proximal portion of the internal claw wider than the primary branch of the external claw. Accessory points on the primary branch of the external claw as long as the branch and turned downwards (Fig. 5 A), the accessory points of the primary branch of the internal claw shorter than the branch. Basal portions of all claws slender, with a septum dividing the primary branch from the rest of the claw. The base of the external claw with a drop-shaped cuticular internal expansion (lunule?), the base of internal claw expanded with lateral filaments. Two long cuticular bars, subdivided into two parts, present under the claws on each of the first three pairs of legs. Differential diagnosis. Thulinius romanoi sp. nov. differs from all other Thulinius species by the unique sculptured cuticle. It is similar to T. augusti and T. stephaniae by the absence of lunules, but the first species has a very thin claw basal tract (in addition to the peculiar presence of subdivided lobes), and the latter has a stronger basal tract and a very marked transverse bar at the base of the claws on the first three pairs of legs. In addition to the sculptured cuticle, T. romanoi sp. nov. differs from T. saltursus and all other Thulinius species by the absence, or not evident, lunule at the base of each external claw and by having a more robust basal tract. A key to the species of Thulinius, excluding T. romanoi sp. nov., can be found in Kaczmarek et al. (2010). Etymology. This species is named after the tardigradologist, and our friend and colleague, the late Frank Romano, whose enthusiasm for tardigrades was infectious. He and his students attended several of the International Symposia on Tardigrades during his tenure as Professor and Chairman of the Department of Biology at Jacksonville State University, Alabama, USA., Published as part of Bertolani, Roberto, Bartels, Paul J., Guidetti, Roberto, Cesari, Michele & Nelson, Diane R., 2014, Aquatic tardigrades in the Great Smoky Mountains National Park, North Carolina and Tennessee, U. S. A., with the description of a new species of Thulinius (Tardigrada, Isohypsibiidae), pp. 524-536 in Zootaxa 3764 (5) on pages 528-533, DOI: 10.11646/zootaxa.3764.5.2, http://zenodo.org/record/230310, {"references":["McInnes, S. 1994. Zoogeographic distribution of terrestrial / freshwater tardigrades from current literature. Journal of Natural History, 28, 257 - 352. http: // dx. doi. org / 10.1080 / 00222939400770131","Bertolani, R., Marley, N. J. & Nelson, D. R. (1999) Re-description of the genus Thulinia (Eutardigrada: Hypsibiidae) and of Thulinia augusti (Murray, 1907) comb. n. Zoologischer Anzeiger, 238, 139 - 145.","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, 699 - 706. http: // dx. doi. org / 10.1111 / j. 1755 - 0998.2009.02538. x","Kaczmarek, L, Bertolani, R. & Nedzynska-Stygar, M. (2010) Thulinius saltursus comb. nov.: a new systematic position for Isohypsibius saltursus Schuster, Toftner & Grigarick, 1978 (Eutardigrada: Hypsibiidae) and a key for the genus Thulinius. Zootaxa, 2483, 58 - 64."]}

As part of the All Taxa Biodiversity Inventory (http://www.dlia.org), an extensive survey of tardigrades has been conducted in the Great Smoky Mountains National Park (GSMNP) in Tennessee and North Carolina, U.S.A., by Bartels and Nelson. Freshwater tardigrades include three species in the aquatic genus Thulinius (Eutardigrada, Isohypsibiidae). A new species, Thulinius romanoi, described from stream sediment, is distinguished from all other congeners by having a sculptured cuticle. In addition, the presence of Thulinius augusti (Murray, 1907) was verified by combined morphological and molecular analysis, and nine specimens of a third species, Thulinius cf. saltursus, were also found. Thulinius augusti is a new record for the United States. Thulinius saltursus (Schuster, ToftnerGrigarick, 1978) was previously recorded in California and Ohio, but our specimens vary slightly in morphology. The list of tardigrades from streams in the GSMNP was updated to a total of 44 species, 22 of which were predominantly or exclusively aquatic.

Diploechiniscus gen. nov. Diagnosis. Echiniscids with dorsal plates I, II, III, IV (II and III paired), transversally subdivided median plates m 1 and m 2 and undivided plate m 3 present; double sculpture in the dorsal plates, represented (under phase contrast) by dark polygonal and white circular grains; ventral plates present, especially evident in the anterior, head region and around the gonopore; supernumerary dorsal-lateral spines present; buccal tube long and narrow, with stylet supports. Orange body, dark-brown eyes. Type species: Echiniscus oihonnae Richters, 1903 Composition: Diploechiniscus oihonnae (Richters, 1903) comb. nov., to date the only species attributable to the new genus. Junior synonym: Echiniscus multispinosus Etymology: from the Greek ������������ς (dipl��os) = double, composed of two parts; referring to the cuticle sculpture, and Echiniscus, the first of the echiniscid genera to be described. Remarks. The echiniscid genera most similar to Diploechiniscus are Testechiniscus, Echiniscus and Bryodelphax. Diploechiniscus is differentiated from Testechiniscus by the presence of double sculpture in the dorsal plates, subdivided dorsal plate m 2 and dorsal plate m 3. It is differentiated from Echiniscus by the presence of black eyes, subdivided dorsal plates m 1 and m 2, double sculpture in the dorsal plates, supernumerary dorsallateral spines, ventral plates and evident stylet supports. From Bryodelphax, Diploechiniscus is differentiated by the presence of black eyes, supernumerary dorsal-lateral spines, dorsal and lateral filaments or spines (apart from filament A), terminal plate notched, and the adults are much larger. The juxtaposition of the four genera into different evolutionary lines within the Echiniscidae was confirmed by 18 S sequences. Diploechiniscus oihonnae (Richters, 1903) comb. nov. Type locality: Merok, Norway Diagnosis. Body colour reddish-brown. Dark brown eye spots. Stylet supports present. Long filaments A, B, C, D and E. Short hooked dorsal-lateral spines B���, C���, D��� and E���. Long filaments C d and short spines D d. Spine B d present or absent. Dorsal plates present: I, paired II and III, and IV, transversally subdivided median plates m 1 and m 2 and median plate m 3 entire. Terminal plate (IV) notched. Double sculpture of the dorsal plates observed under light microscopy. Faint ventral plates present, with those at the anterior and posterior more clearly visible. Sensory spine on leg I and papilla on leg IV, present. Lateral leg plates, present. Dentate collar on leg IV, present. Females and males present, with gonopores typical of the echiniscid form. Re-description of the species (from the original description and from re-examined specimens collected in Fors��, Norway; Sierra de Urbion, Spain; Caldas das Taipas, Vilar Formoso, Castro Laboreiro and Moita do Conqueiro, Portugal). Body colour orange. Eye spots simple and dark brown. Buccal cirri long, clavae large. Stylet supports present (sometimes difficult to observe in older slides). Dorsal plates present, all (except neck plate) characterized by double sculpture, which appears as dark, regular polygonal grains under white circular grains when viewed with phase contrast. Dark grains are separated by thin, white region from neighbours (normally groups of six); white grains of various sizes, never overlaping dark grains, and irregularly distributed. Cephalic plate unpaired, with median depression to the anterior margin; fine anterior sculpture and larger posterior double sculpture. Neck plate, a long transverse and relatively thin band, anterior and posterior region unsculptured and fine, dark grains in the middle. Dorsal segmental plates: plate I (or scapular plate) entire, with two sculptured small lateral plates exhibiting fine, dark grains; plates II and III paired and characterised by an unsculptured transverse band, and plate IV (or terminal plate), entire but faceted and notched (not obvious in older specimens). Median intersegmental plates: plate m 1, transversally subdivided, anterior region formed of a large, flat and thin rectangle not always obvious due to overlapping scapular plate; plate m 2, transversally subdivided and with an unsculptured transverse band, plate appears as two obtuse angle isosceles triangles joined by their larger side; and m 3, entire, small and not obvious but with double sculpture. Lateral intersegmental plates are difficult to identify, though unsculptured spaces exist at la 2 and la 3. Long filaments A, B, C, D and E, sometimes barbed. Short hooked dorsal-lateral spines B���, C���, D��� and E���. Lateral spine E���, simple or double. B d variable as long spine, very short spur, or absent and can be present on one or both sides of plate II. Long filaments C d and short spines D d. Ventral sculpture present as fine granulation, with clearly visible head plate and posterior plates beside gonopore. Leg plates present laterally, with dark granular sculpture. Spiniform papilla present on leg I; papilla on leg IV with rounded tip. Hooked spurs on all internal claws, external claws I���III smooth, occasionally one or two short right-angled spurs on the leg IV. Dentate collar variable, comprised of six to 13 triangular teeth, some irregularly bifurcated. Gonopore; a short tube in the males, and rosette in the females. The geographical distribution of E. oihonnae includes: Portugal, Switzerland, Northern Europe (including polar islands), U.S.A., Canada, Australia (Ramazzotti & Maucci 1983); Japan (Mathews, 1937); Kuril Islands, Far East Russia (Dudichev & Biserov 2000). Most of the non-European citations require confirmation, as for example, Murray (1910) was doubtful about his identification of Australian and Canadian specimens, and the Californian specimens, initially assigned to E. oihonnae, were revised as T. laterculus (Schuster, Grigarick & Toftner, 1980). Acknowledgements This study was partially supported by the Funda����o para a Ci��ncia e a Tecnologia, Portugal, with a grant (BD/ 39234 / 2007) to the first author and by the program Pest-OE/MAR/UI0331/ 2011 to the research of the second author, and also by the European Distributed Institute of Taxonomy (EDIT) within the program ATBI: All Taxa Biodiversity Inventories in the Gemer Area, Slovakia. The research is also part of the project MoDNA supported by Fondazione Cassa di Risparmio di Modena (Italy) and the University of Modena and Reggio Emilia (Modena, Italy). The authors wish to thank Dr. C��sar Garcia (Botanical Garden, Lisbon) for providing the moss samples from the Portuguese locality of Moita do Conqueiro, and Museo Civico di Storia Naturale di Verona for the availability of the slides of the Maucci collection. They also wish to thank Sandra McInnes, of the British Antarctic Survey, for her critical support and the English revision. The authors are also thankful to the two anonymous referees for their helpful comments and suggestions., Published as part of Vicente, Filipe, Fontoura, Paulo, Cesari, Michele, Rebecchi, Lorena, Guidetti, Roberto, Serrano, Artur & Bertolani, Roberto, 2013, Integrative taxonomy allows the identification of synonymous species and the erection of a new genus of Echiniscidae (Tardigrada, Heterotardigrada), pp. 557-572 in Zootaxa 3613 (6) on pages 569-570, DOI: 10.11646/zootaxa.3613.6.3, http://zenodo.org/record/220090

Vicente, Filipe, Fontoura, Paulo, Cesari, Michele, Rebecchi, Lorena, Guidetti, Roberto, Serrano, Artur, Bertolani, Roberto (2013): Integrative taxonomy allows the identification of synonymous species and the erection of a new genus of Echiniscidae (Tardigrada, Heterotardigrada). Zootaxa 3613 (6): 557-572, DOI: http://dx.doi.org/10.11646/zootaxa.3613.6.3

Marchioro, Trevor, Rebecchi, Lorena, Cesari, Michele, Hansen, Jesper Guldberg, Viotti, Giulia, Guidetti, Roberto (2013): Somatic musculature of Tardigrada: phylogenetic signal and metameric patterns. Zoological Journal of the Linnean Society 169 (3): 580-603, DOI: 10.1111/zoj.12079, URL: http://dx.doi.org/10.1111/zoj.12079

Guil, Noemi, Machordom, Annie, Guidetti, Roberto (2013): High level of phenotypic homoplasy amongst eutardigrades (Tardigrada) based on morphological and total evidence phylogenetic analyses. Zoological Journal of the Linnean Society 169 (1): 1-26, DOI: 10.1111/zoj.12046, URL: http://dx.doi.org/10.1111/zoj.12046

Macrobiotus kristenseni sp. nov. (Figures 1–5 and Table 2) Notes: bl = body length; btl = buccal tube length; btd = diameter of the buccal tube; vl = ventral lamina length; ssi = insertion of stylet supports; plr = macroplacoid row length; 1 ◦ pl = first macroplacoid length; 2 ◦ pl = second macroplacoid length; mpl = microplacoid length; pb1-4 = length of primary branch of the claw on first-fourth leg pair; sb1-4 length of secondary the claw on first-fourth leg pair; pt = index pt (length of a structure / buccal tube length ∗ 100; Pilato, 1981); min = minimum; max = maximum; SD = standard deviation; c.v. = coefficient of variation; n = number of considered specimens. Diagnosis Size medium. Cuticle with pearls (pores). Eye-spots present. Buccal tube of average width. Buccal armature with two bands of teeth and transverse ridges. Bulb with two macroplacoids and small microplacoid. Long Y-shaped claws with two short accessory points on the main branch, lunule smooth or weakly crenate in the hind legs. Free eggs with processes in shape of elongated cones with truncated and enlarged apexes. Limnoterrestrial. Description Size range: 247–544 µm long. Colourless, yellowish, greyish or light brown, sometimes with transverse bands with brown pigment. Eye-spots present in all specimens observed (Figure 1), eye-spot colour not detected. Cuticle smooth under LM, except small granulations on outer part of legs. Cuticular pores rounded or ovate approximately 1 µm in diameter (Figure 2), generally with thickened edges, irregularly distributed on the body surface (dorsally and ventrally). Near the anterior end, a ring of pores with evident thickened edges and placed in a regular manner. Ten small peribuccal lamellae surround the mouth opening, followed by six buccal sensory lobes (Figure 3). Buccal armature, poorly distinguishable under LM, formed by an anterior band of small teeth of variable thickness at the base of the buccal lamellae, sometimes reduced to a single row of teeth (Figures 1 and 3); a posterior band of larger teeth, sometimes in a single row, followed by three dorsal and three ventral fine transverse ridges (Figures 1 and 3). Ventral transversal ridges clearly smaller than the dorsal, lateral ridges larger than the middle ones (Figure 1). Long ventral strengthening bar, about 65% of buccal tube total length, with an anterior crest on its ventral margin (Figure 1). Stylet support insertion point posteriorly placed, at about 75% the distance of the buccal tube from the anterior margin of the stylet sheaths. Anterior end of the piercing stylet with external edge straight and serrate, and internal edge curved and smooth (Figure 3). Pharyngeal bulb oval to circular, with well-developed pharyngeal apophyses. Two elongate macroplacoids and a small elongate microplacoid close to the second macroplacoid. First macroplacoid – the longest – with a small medial constriction, second macroplacoid quadrangular or circular (Figure 1). Slanting pharyngeal bars present between the apophyses and the first macroplacoids (Figure 1). Long double-claws. Main branch long with two fine accessory points, not reaching the distal bend of the main branch, and not clearly visible at LM (Figure 2). Secondary branch long and evidently bended. Short and large common tract, without distal part, and with an evident stalk connecting the claw to the lunule (Figure 2). Hind claws larger than the others. Lunules smooth in legs I–III, larger and with weakly crenate edges in the hind legs (Figure 2). Measurements of different animal cuticular structures, pt values and a descriptive statistical analysis are given in Table 2. Ornamented round egg laid free (Figures 4 and 5). Egg ornamentation formed by processes in the shape of elongated empty cones (height 6.2–12.7 µm, diameter of their base 2.0–3.3 µm) with truncated and enlarged apexes (diameter less than 2 µm) (Figures 4, 5). Several processes with a long thick filament developing from the distal disk (Figures 4, 5). Ornamentations with smooth surface frequently ringed. Diameter of eggs without processes 73.0–96.3 µm, diameter of eggs with processes 94.0–125.0 µm. Number of processes present on a diameter 10–13. Chorion smooth; in some eggs a fine granulation and very small scattered pores in between processes visible by LM (Figure 4). Three embryonated eggs were found (Figure 4). In addition to the typical eggs described above, few eggs present slightly higher and more flexible processes internally septated and with a smaller base (Figures 4, 5). Type locality South America, Argentina, La Pampa Province, city of Santa Rosa (36 ◦ 39 ′ S, 64 ◦ 17 ′ W) at 177 m above sea level, epiphytic lichens. Type material Holotype (MLP-Oi 3726), four paratypes (MLP-Oi 3727) and three eggs are housed at the Museum of Natural Sciences, National University of La Plata (Argentina), and 32 paratypes are housed in J. Peluffo’s personal collection in the Department of Natural Sciences at the National University of La Pampa (Argentina). The voucher specimens (two pictures of the hologenophore animals, 16 cryoconserved paragenophore animals, eight paratypes / paragenophore animals and eggs) and a fragment of the lichen are deposited in the MOD:BRT collection of the Department of Life Sciences (University of Modena and Reggio Emilia, Italy) registered in the Registry of Biological Repositories (www.biorepositories.org). Additional material South America, Argentina, La Pampa Province, city of General Pico at 143 m above sea level (125 km northeast of Santa Rosa) and Natural Reserve Parque Luro (35 km south of Santa Rosa, caldén forest), in all cases on epiphytic lichens. Etymology The name is dedicated in honour of Prof. Reinhardt M. Kristensen, of the Invertebrate Department of The Natural History Museum of Denmark of the University of Copenhagen. Remarks Under SEM, the general surface of the cuticle appears slightly wrinkled (Figure 2). It should be verified whether this character is an effect of SEM preparation. An egg observed by SEM had processes with filamentous strips on their surface (Figure 5). These strips seem to be present on the same side of all processes: it is not clear whether they are artefacts or real morphological structures. Where M. kristenseni sp. nov. appears in a sample, it is either the only or the dominant tardigrade species. In the latter case, it is accompanied by one or more of the following species: Echiniscus rufoviridis du Bois Raymond-Marcus, 1944; Milnesium cf. tardigradum ; Ramazzottius cf. oberhaeuseri. The phorophyte of the lichen communities in which it was recorded more frequently is the autochthonous tree species Prosopis caldenia (Burk.). Taxonomic remarks. Macrobiotus kristenseni sp. nov. is similar to species of the M. hufelandi group. Among these species, Macrobiotus sandrae Bertolani and Rebecchi, 1993 is the most similar, sharing a weak buccal armature, reduced ventral transversal crests, and indented lunule only on the fourth leg pair, but it differs from M. kristenseni sp. nov. by longer claw accessory points and larger dorsal transversal crests. Macrobiotus kristenseni sp. nov. differs from all the M. hufelandi group of species in that the shape of the egg processes is different from any other egg of the species group. The most similar eggs are those of Macrobiotus recens Cuénot, 1932, which share similar thin, long, conical processes with a truncated apex but in M. recens an evident series of small indentations around the base of the egg processes is present (Pilato and Bertolani 2004). Molecular results The molecular phylogeny based on the 18S rRNA gene (897 bp) indicates that M. kristenseni sp. nov. does not share its 18S rRNA sequence with any other Macrobiotus species sequenced so far (Figure 6), but it is in a sister-group relationship with Macrobiotus sapiens Binda and Pilato, 1984 within a well-supported clade grouping all the M. hufelandi group of species (i.e. M. sapiens, Macrobiotus hufelandi C.A.S. Schultze, 1834, Macrobiotus polonicus Pilato, Kaczmarek, Michalczyk and Lisi, 2003, Macrobiorus vladimiri Bertolani, Biserov, Rebecchi and Cesari, 2011). Acronyms as in Table 1. Molecular analysis of the mitochondrial DNA cox1 gene was carried out on sequences of 491–624 bp. Two different sequences (i.e. two haplotypes) are present in the five specimens analysed: three specimens are of one haplotype, two specimens are of the other (Table 3; Figure 7). The two haplotypes are very similar and the intraspecific genetic distance among M. kristenseni sp. nov. specimens is low (1.4–1.5%; Table 3). Mean K2P distances between specimens pertaining to different species are very high (18.2–33.4%; Table 4). In particular, among the considered species, M. kristenseni sp. nov. has the greatest genetic difference with respect to the other M. hufelandi group of species (23.1–33.4%), while the difference among the considered species excluding M. kristenseni sp. nov. is 18.2–24.4%. The neighbour-joining dendrogram based on cox1 sequences confirms these differences between the two M. kristenseni sp. nov. haplotypes, and the differences among this species and the other species of M. hufelandi group (Figure 7).

Macrobiotus kristenseni sp. nov. (Figures 1–5 and Table 2) Notes: bl = body length; btl = buccal tube length; btd = diameter of the buccal tube; vl = ventral lamina length; ssi = insertion of stylet supports; plr = macroplacoid row length; 1 ◦ pl = first macroplacoid length; 2 ◦ pl = second macroplacoid length; mpl = microplacoid length; pb1-4 = length of primary branch of the claw on first-fourth leg pair; sb1-4 length of secondary the claw on first-fourth leg pair; pt = index pt (length of a structure / buccal tube length ∗ 100; Pilato, 1981); min = minimum; max = maximum; SD = standard deviation; c.v. = coefficient of variation; n = number of considered specimens. Diagnosis Size medium. Cuticle with pearls (pores). Eye-spots present. Buccal tube of average width. Buccal armature with two bands of teeth and transverse ridges. Bulb with two macroplacoids and small microplacoid. Long Y-shaped claws with two short accessory points on the main branch, lunule smooth or weakly crenate in the hind legs. Free eggs with processes in shape of elongated cones with truncated and enlarged apexes. Limnoterrestrial. Description Size range: 247–544 µm long. Colourless, yellowish, greyish or light brown, sometimes with transverse bands with brown pigment. Eye-spots present in all specimens observed (Figure 1), eye-spot colour not detected. Cuticle smooth under LM, except small granulations on outer part of legs. Cuticular pores rounded or ovate approximately 1 µm in diameter (Figure 2), generally with thickened edges, irregularly distributed on the body surface (dorsally and ventrally). Near the anterior end, a ring of pores with evident thickened edges and placed in a regular manner. Ten small peribuccal lamellae surround the mouth opening, followed by six buccal sensory lobes (Figure 3). Buccal armature, poorly distinguishable under LM, formed by an anterior band of small teeth of variable thickness at the base of the buccal lamellae, sometimes reduced to a single row of teeth (Figures 1 and 3); a posterior band of larger teeth, sometimes in a single row, followed by three dorsal and three ventral fine transverse ridges (Figures 1 and 3). Ventral transversal ridges clearly smaller than the dorsal, lateral ridges larger than the middle ones (Figure 1). Long ventral strengthening bar, about 65% of buccal tube total length, with an anterior crest on its ventral margin (Figure 1). Stylet support insertion point posteriorly placed, at about 75% the distance of the buccal tube from the anterior margin of the stylet sheaths. Anterior end of the piercing stylet with external edge straight and serrate, and internal edge curved and smooth (Figure 3). Pharyngeal bulb oval to circular, with well-developed pharyngeal apophyses. Two elongate macroplacoids and a small elongate microplacoid close to the second macroplacoid. First macroplacoid – the longest – with a small medial constriction, second macroplacoid quadrangular or circular (Figure 1). Slanting pharyngeal bars present between the apophyses and the first macroplacoids (Figure 1). Long double-claws. Main branch long with two fine accessory points, not reaching the distal bend of the main branch, and not clearly visible at LM (Figure 2). Secondary branch long and evidently bended. Short and large common tract, without distal part, and with an evident stalk connecting the claw to the lunule (Figure 2). Hind claws larger than the others. Lunules smooth in legs I–III, larger and with weakly crenate edges in the hind legs (Figure 2). Measurements of different animal cuticular structures, pt values and a descriptive statistical analysis are given in Table 2. Ornamented round egg laid free (Figures 4 and 5). Egg ornamentation formed by processes in the shape of elongated empty cones (height 6.2–12.7 µm, diameter of their base 2.0–3.3 µm) with truncated and enlarged apexes (diameter less than 2 µm) (Figures 4, 5). Several processes with a long thick filament developing from the distal disk (Figures 4, 5). Ornamentations with smooth surface frequently ringed. Diameter of eggs without processes 73.0–96.3 µm, diameter of eggs with processes 94.0–125.0 µm. Number of processes present on a diameter 10–13. Chorion smooth; in some eggs a fine granulation and very small scattered pores in between processes visible by LM (Figure 4). Three embryonated eggs were found (Figure 4). In addition to the typical eggs described above, few eggs present slightly higher and more flexible processes internally septated and with a smaller base (Figures 4, 5). Type locality South America, Argentina, La Pampa Province, city of Santa Rosa (36 ◦ 39 ′ S, 64 ◦ 17 ′ W) at 177 m above sea level, epiphytic lichens. Type material Holotype (MLP-Oi 3726), four paratypes (MLP-Oi 3727) and three eggs are housed at the Museum of Natural Sciences, National University of La Plata (Argentina), and 32 paratypes are housed in J. Peluffo’s personal collection in the Department of Natural Sciences at the National University of La Pampa (Argentina). The voucher specimens (two pictures of the hologenophore animals, 16 cryoconserved paragenophore animals, eight paratypes / paragenophore animals and eggs) and a fragment of the lichen are deposited in the MOD:BRT collection of the Department of Life Sciences (University of Modena and Reggio Emilia, Italy) registered in the Registry of Biological Repositories (www.biorepositories.org). Additional material South America, Argentina, La Pampa Province, city of General Pico at 143 m above sea level (125 km northeast of Santa Rosa) and Natural Reserve Parque Luro (35 km south of Santa Rosa, caldén forest), in all cases on epiphytic lichens. Etymology The name is dedicated in honour of Prof. Reinhardt M. Kristensen, of the Invertebrate Department of The Natural History Museum of Denmark of the University of Copenhagen. Remarks Under SEM, the general surface of the cuticle appears slightly wrinkled (Figure 2). It should be verified whether this character is an effect of SEM preparation. An egg observed by SEM had processes with filamentous strips on their surface (Figure 5). These strips seem to be present on the same side of all processes: it is not clear whether they are artefacts or real morphological structures. Where M. kristenseni sp. nov. appears in a sample, it is either the only or the dominant tardigrade species. In the latter case, it is accompanied by one or more of the following species: Echiniscus rufoviridis du Bois Raymond-Marcus, 1944; Milnesium cf. tardigradum ; Ramazzottius cf. oberhaeuseri. The phorophyte of the lichen communities in which it was recorded more frequently is the autochthonous tree species Prosopis caldenia (Burk.). Taxonomic remarks. Macrobiotus kristenseni sp. nov. is similar to species of the M. hufelandi group. Among these species, Macrobiotus sandrae Bertolani and Rebecchi, 1993 is the most similar, sharing a weak buccal armature, reduced ventral transversal crests, and indented lunule only on the fourth leg pair, but it differs from M. kristenseni sp. nov. by longer claw accessory points and larger dorsal transversal crests. Macrobiotus kristenseni sp. nov. differs from all the M. hufelandi group of species in that the shape of the egg processes is different from any other egg of the species group. The most similar eggs are those of Macrobiotus recens Cuénot, 1932, which share similar thin, long, conical processes with a truncated apex but in M. recens an evident series of small indentations around the base of the egg processes is present (Pilato and Bertolani 2004). Molecular results The molecular phylogeny based on the 18S rRNA gene (897 bp) indicates that M. kristenseni sp. nov. does not share its 18S rRNA sequence with any other Macrobiotus species sequenced so far (Figure 6), but it is in a sister-group relationship with Macrobiotus sapiens Binda and Pilato, 1984 within a well-supported clade grouping all the M. hufelandi group of species (i.e. M. sapiens, Macrobiotus hufelandi C.A.S. Schultze, 1834, Macrobiotus polonicus Pilato, Kaczmarek, Michalczyk and Lisi, 2003, Macrobiorus vladimiri Bertolani, Biserov, Rebecchi and Cesari, 2011). Acronyms as in Table 1. Molecular analysis of the mitochondrial DNA cox1 gene was carried out on sequences of 491–624 bp. Two different sequences (i.e. two haplotypes) are present in the five specimens analysed: three specimens are of one haplotype, two specimens are of the other (Table 3; Figure 7). The two haplotypes are very similar and the intraspecific genetic distance among M. kristenseni sp. nov. specimens is low (1.4–1.5%; Table 3). Mean K2P distances between specimens pertaining to different species are very high (18.2–33.4%; Table 4). In particular, among the considered species, M. kristenseni sp. nov. has the greatest genetic difference with respect to the other M. hufelandi group of species (23.1–33.4%), while the difference among the considered species excluding M. kristenseni sp. nov. is 18.2–24.4%. The neighbour-joining dendrogram based on cox1 sequences confirms these differences between the two M. kristenseni sp. nov. haplotypes, and the differences among this species and the other species of M. hufelandi group (Figure 7)., Published as part of Guidetti, Roberto, Peluffo, Julio Ricardo, Rocha, Alejandra Mariana, Cesari, Michele & Peluffo, María Cristina Moly de, 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), pp. 2409-2426 in Journal of Natural History 47 (37 - 38) on pages 2413-2422, DOI: 10.1080/00222933.2013.800610, http://zenodo.org/record/5197685, {"references":["Pilato G. 1981. Analisi di nuovi caratteri nello studio degli Eutardigradi. Animalia. 8: 51 - 57.","Bertolani R, Rebecchi L. 1993. A revision of the Macrobiotus hufelandi group (Tardigrada, Macrobiotidae), with some observations on the taxonomic characters of eutardigrades. Zool. Scripta. 22: 127 - 152.","Pilato G, Bertolani R. 2004. Macrobiotus dariae sp. n., a new species of eutardigrade (Eutardigrada, Macrobiotidae) from Cyprus. Zootaxa. 638: 1 - 7.","Cesari M, Giovannini 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."]}

Macrobiotus caelicola (Figures 4F, 5E, F) Buccal armature consists of a posterior row (two rows in a large specimen) of quite large, rounded teeth. At the position of the lateral transverse crests, two rounded teeth occur at both dorsal and ventral locations; central transverse crest is absent. Internal surface of the anterior portion of the buccal tube has a large dorsal tooth (not cited in the original description; Kathman 1990) (Figure 4E, F). Slanting cuticular bars (between apophyses and first macroplacoid) can be observed in the pharynx., Published as part of Guil, Noemí, Guidetti, Roberto & Machordom, Annie, 2007, Observations on the '' tenuis group' ' (Eutardigrada, Macrobiotidae) and description of a new Macrobiotus species, pp. 2741-2755 in Journal of Natural History 41 (41 - 44) on page 2752, DOI: 10.1080/00222930701742637, http://zenodo.org/record/5233498, {"references":["Kathman RD. 1990. Some tardigrades from Colorado, with a description of a new species of Macrobiotus (Macrobiotidae: Eutardigrada). Proceedings of the Biological Society of Washington 103: 300 - 303."]}

Macrobiotus ariekammensis (Figures 4C, 5B, D) Characters previously not cited in descriptions of M. ariekammensis and/or M. adelges (Węglarska 1965; Dastych 1977, 1985) are: cuticular pearls over the entire surface of the animal, larger pearls in anterior (mainly around the mouth opening) and caudal dorsal regions; buccal tube with a clear bend in an anterior position (immediately behind the mouth opening). At this level, the animal has a large dorsal tooth on the internal surface of the buccal tube (Figure 5B, D); in lateral view, the ventral portion of the buccal tube is shorter and ends at the level of the stylet sheaths; ventral lamina has a wide ventral crest characterized by two lobes in lateral view (Figure 5D). Contrary to that reported by Dastych (1985), the buccal armature contains a posterior band of small teeth (transverse crests absent)., Published as part of Guil, Noemí, Guidetti, Roberto & Machordom, Annie, 2007, Observations on the '' tenuis group' ' (Eutardigrada, Macrobiotidae) and description of a new Macrobiotus species, pp. 2741-2755 in Journal of Natural History 41 (41 - 44) on page 2750, DOI: 10.1080/00222930701742637, http://zenodo.org/record/5233498, {"references":["Weglarska B. 1965. Die Tardigraden (Tardigrada) Spitzbergens. Acta Zoologica Cracoviensia 11: 43 - 51.","Dastych H. 1977. Macrobiotus adelges sp. nov., a new species of Tardigrada from Poland. Bulletin de l'Academie Polonaise des Sciences, Series des Sciences Biologiques (Cl 2) 25: 667 - 669.","Dastych H. 1985. West Spitsbergen Tardigrada. Acta Zoologica Cracoviensia 28: 169 - 214."]}

Macrobiotus hystricogenitus (Figure 4E) Contrary to the description by Maucci (1978), the buccal armature comprises a posterior band of small teeth (due to the position of the buccal tube, it was not possible to detect the presence of transverse crests)., Published as part of Guil, Noemí, Guidetti, Roberto & Machordom, Annie, 2007, Observations on the '' tenuis group' ' (Eutardigrada, Macrobiotidae) and description of a new Macrobiotus species, pp. 2741-2755 in Journal of Natural History 41 (41 - 44) on page 2752, DOI: 10.1080/00222930701742637, http://zenodo.org/record/5233498, {"references":["Maucci W. 1978. Tardigradi muscicoli della Turchia (Terzo contributo). Bollettino del Museo Civico di Storia Naturale di Verona 5: 111 - 140."]}

Macrobiotus hyperonyx (Figures 4D, 5C) Buccal tube has a clear bend in its anterior portion (just after the mouth opening). At this level, the animal has a large dorsal tooth on the inner surface of the buccal tube (Figure 5A, C), which was not cited in the original description (Maucci 1982). In lateral view, the ventral portion of the buccal tube is shorter and ends at the stylet sheaths. Unlike the original description, the buccal armature comprises a posterior band of small teeth (transverse crests absent)., Published as part of Guil, Noemí, Guidetti, Roberto & Machordom, Annie, 2007, Observations on the '' tenuis group' ' (Eutardigrada, Macrobiotidae) and description of a new Macrobiotus species, pp. 2741-2755 in Journal of Natural History 41 (41 - 44) on pages 2750-2752, DOI: 10.1080/00222930701742637, http://zenodo.org/record/5233498, {"references":["Maucci W. 1982. Sulla presenza in Italia di Cornechiniscus holmeni (Petersen, 1951) e descrizione di Macrobiotus hyperonyx sp. nov. (Tardigrada). Bollettino del Museo Civico di Storia Naturale di Verona IX: 175 - 179."]}

Minibiotus gumersindoi n. sp. Diagnosis Cuticle with transversal bands of large and round pores, a very large round pore close to the end of each leg; granulation on legs not visible; three round macroplacoids of the same size and a solid and distinct microplacoid; slender claws with short but evident accessory points and smooth lunules. Description Body length 162.0 m (Tables 1–2), colourless. Eyes not visible, probably lost due to mounting medium action (see Remarks). Cuticle with transverse bands of round and large pores around the body (Figure 1). Pores larger on the head region. A very large and round pore present close to the claws on external side of all legs (pore diameter = 2.0% of body length; Figures 1, 2 A; Tables 1–2). In some legs, three or five smaller pores forming a triangle or pentagon figure present over the large round pore (Figure 2 A). Buccal armature and transversal ridges not visible at light microscope. Buccal tube long (19.8 µm) and narrow (1.2 µm) (Tables 1–2) and with double curvature (Figure 2 B). Stylet support inserted at 55.0% of buccal tube length (Tables 1–2). Pharyngeal bulb round (Tables 1–2) with large, round apophyses, three round macroplacoids of similar size (Figure 2 B) and solid, and small but distinct microplacoid (Figures 1, 2 B). Macroplacoid row: 5.4 m (27.5 % of buccal tube length; Table 1). Claws slender (main branch of fourth pair of claws 4.9 m long) with long secondary branches (Figure 2 C). Short but well developed accessory points on main branches present (Figure 2 C). Lunules small, smooth and thin in all claws (Figure 2 C). Eggs unknown. Etymology The species is named in memory of Mr. Gumersindo Guil Valverde, Dr. Guil’s grandfather. Type locality Moss on rock, in oak forest at 1098 m a.s.l. in Bustarviejo Valley (Madrid, Spain; UTM 30 T0437017­ 4521195). Repositories The holotype and paratype are deposited in the non­insect invertebrate collection of National Museum of Natural Science of Madrid (Consejo Superior de Investigaciones Científicas, CSIC) (collection number: 23.00/ 1). Remarks Cuticle transverse bands are not well identifiable in the paratype and in some specimens of west Madrid and Canary Islands. Eyes are visible in specimens of Minibiotus gumersindoi n. sp. from west Madrid and Canary Islands. Differential diagnosis Minibiotus gumersindoi n. sp. differs from all the other species of the genus for the presence of a very large pore in the external distal part of the leg and for a set of large pores in the proximal part of the leg. In particular Minibiotus gumersindoi n. sp. differs from other similar species (Table 3) for: ­ the presence of large and almost regular rounded pores: irregular in M. bisoctus (Horning, Schuster & Grigarick), M. ethelae Claxton, M. furcatus (Ehrenberg), M. stuckenbergi (Dastych, Ryan & Watkins), and M. vinciguerrae Binda & Pilato, and small in M. keppelensis Claxton, M. poricinctus Claxton and M. ramazzottii Binda & Pilato; ­ the absence of granulation on legs: present in M. bisoctus (Horning, Schuster & Grigarick), M. keppelensis Claxton, M. poricinctus Claxton, M. stuckenbergi (Dastych, Ryan & Watkins), M. vinciguerrae Binda & Pilato, and M. weinerorum (Dastych); ­ the absence of cuticle thickened in caudal region: present in M. ethelae Claxton; ­ the presence of three macroplacoids; two in M. scopulus Claxton; ­ the presence of smooth lunules on the fourth leg pair: with teeth in M. stuckenbergi (Dastych, Ryan & Watkins). Habitat In Cercedilla, specimens were found in rock mosses in a xerophilus landscape (xerophilus bushes: thyme, rosemary and broom) at 1228 m a.s.l.. In Embalse de la Jarosa the new species has been found in rock mosses in a pine forest (of Pinus nigra Arnold) at 1107 m a.s.l. and in the Canary Islands in rock mosses in a pine forest (of Pinus canariensis Smith) at 1460 m a.s.l. Population variability We consider the specimens collected in the different localities belonging to the same species due to the well defined morphological characters shared by these specimens. Due to the relatively small number of specimens collected in the different localities, we did not performed a statistical analyses of the morphometric data. Anyway, we report the morphometric differences identified in the populations. Morphometric differences appear when the Madrid populations (Table 1 from M 1 to M 2, type locality population, and from M 3 to M 7, west Madrid populations) and the Caldera de Taburiente (Canary Islands) population (Table 1 from CT 1 to CT 9) are compared. Body length, stylet support insertion length and secondary branch of claws in fourth pair of legs are larger in the Caldera de Taburiente (Canary Islands) population than in the Madrid (type locality) and the west Madrid populations (Table 2), while main branch of claws in fourth pair of legs is larger in the Madrid populations than in the Caldera de Taburiente (Canary Islands) population (Table 2). The Caldera de Taburiente population (Canary Islands) (from pine forest) has the bigger distal leg pore diameters, followed by the west Madrid population (from xerophilous and pine forest) and the type locality population (from oak forest) (Table 2). Buccal tube length and body pores diameters are larger in the type locality population than in the west Madrid and the Caldera de Taburiente (Canary Islands) populations, where their values are similar. Body length and pharyngeal width are shorter in the type locality population than in the others (Table 2). Other species of tardigrades There is no other tardigrade species that occurs always in all the four localities where we have found Minibiotus gumersindoi n. sp. Echiniscus trisetosus Cuénot is present in three of the four localities (Bustarviejo valley, Cercedilla and Caldera de Taburiente). We present other tardigrade species found together with the new species in Table 4, where information is given on which species are new records for Madrid, Canary Islands and/or the Iberian Peninsula., Published as part of Guil, Noemi & Guidetti, Roberto, 2005, A new species of Tardigrada (Eutardigrada: Macrobiotidae) from Iberian Peninsula and Canary Islands (Spain), pp. 1-11 in Zootaxa 889 on pages 3-8, DOI: 10.5281/zenodo.170921, {"references":["Claxton, S. (1998) A revision of the genus Minibiotus (Tardigrada: Macrobiotidae) with description of eleven new species from Australia. Records of the Australian Museum, 50 (2), 125 - 160."]}

Minibiotus gumersindoi n. sp. Diagnosis Cuticle with transversal bands of large and round pores, a very large round pore close to the end of each leg; granulation on legs not visible; three round macroplacoids of the same size and a solid and distinct microplacoid; slender claws with short but evident accessory points and smooth lunules. Description Body length 162.0 m (Tables 1–2), colourless. Eyes not visible, probably lost due to mounting medium action (see Remarks). Cuticle with transverse bands of round and large pores around the body (Figure 1). Pores larger on the head region. A very large and round pore present close to the claws on external side of all legs (pore diameter = 2.0% of body length; Figures 1, 2 A; Tables 1–2). In some legs, three or five smaller pores forming a triangle or pentagon figure present over the large round pore (Figure 2 A). Buccal armature and transversal ridges not visible at light microscope. Buccal tube long (19.8 µm) and narrow (1.2 µm) (Tables 1–2) and with double curvature (Figure 2 B). Stylet support inserted at 55.0% of buccal tube length (Tables 1–2). Pharyngeal bulb round (Tables 1–2) with large, round apophyses, three round macroplacoids of similar size (Figure 2 B) and solid, and small but distinct microplacoid (Figures 1, 2 B). Macroplacoid row: 5.4 m (27.5 % of buccal tube length; Table 1). Claws slender (main branch of fourth pair of claws 4.9 m long) with long secondary branches (Figure 2 C). Short but well developed accessory points on main branches present (Figure 2 C). Lunules small, smooth and thin in all claws (Figure 2 C). Eggs unknown. Etymology The species is named in memory of Mr. Gumersindo Guil Valverde, Dr. Guil’s grandfather. Type locality Moss on rock, in oak forest at 1098 m a.s.l. in Bustarviejo Valley (Madrid, Spain; UTM 30 T0437017­ 4521195). Repositories The holotype and paratype are deposited in the non­insect invertebrate collection of National Museum of Natural Science of Madrid (Consejo Superior de Investigaciones Científicas, CSIC) (collection number: 23.00/ 1). Remarks Cuticle transverse bands are not well identifiable in the paratype and in some specimens of west Madrid and Canary Islands. Eyes are visible in specimens of Minibiotus gumersindoi n. sp. from west Madrid and Canary Islands. Differential diagnosis Minibiotus gumersindoi n. sp. differs from all the other species of the genus for the presence of a very large pore in the external distal part of the leg and for a set of large pores in the proximal part of the leg. In particular Minibiotus gumersindoi n. sp. differs from other similar species (Table 3) for: ­ the presence of large and almost regular rounded pores: irregular in M. bisoctus (Horning, Schuster & Grigarick), M. ethelae Claxton, M. furcatus (Ehrenberg), M. stuckenbergi (Dastych, Ryan & Watkins), and M. vinciguerrae Binda & Pilato, and small in M. keppelensis Claxton, M. poricinctus Claxton and M. ramazzottii Binda & Pilato; ­ the absence of granulation on legs: present in M. bisoctus (Horning, Schuster & Grigarick), M. keppelensis Claxton, M. poricinctus Claxton, M. stuckenbergi (Dastych, Ryan & Watkins), M. vinciguerrae Binda & Pilato, and M. weinerorum (Dastych); ­ the absence of cuticle thickened in caudal region: present in M. ethelae Claxton; ­ the presence of three macroplacoids; two in M. scopulus Claxton; ­ the presence of smooth lunules on the fourth leg pair: with teeth in M. stuckenbergi (Dastych, Ryan & Watkins). Habitat In Cercedilla, specimens were found in rock mosses in a xerophilus landscape (xerophilus bushes: thyme, rosemary and broom) at 1228 m a.s.l.. In Embalse de la Jarosa the new species has been found in rock mosses in a pine forest (of Pinus nigra Arnold) at 1107 m a.s.l. and in the Canary Islands in rock mosses in a pine forest (of Pinus canariensis Smith) at 1460 m a.s.l. Population variability We consider the specimens collected in the different localities belonging to the same species due to the well defined morphological characters shared by these specimens. Due to the relatively small number of specimens collected in the different localities, we did not performed a statistical analyses of the morphometric data. Anyway, we report the morphometric differences identified in the populations. Morphometric differences appear when the Madrid populations (Table 1 from M 1 to M 2, type locality population, and from M 3 to M 7, west Madrid populations) and the Caldera de Taburiente (Canary Islands) population (Table 1 from CT 1 to CT 9) are compared. Body length, stylet support insertion length and secondary branch of claws in fourth pair of legs are larger in the Caldera de Taburiente (Canary Islands) population than in the Madrid (type locality) and the west Madrid populations (Table 2), while main branch of claws in fourth pair of legs is larger in the Madrid populations than in the Caldera de Taburiente (Canary Islands) population (Table 2). The Caldera de Taburiente population (Canary Islands) (from pine forest) has the bigger distal leg pore diameters, followed by the west Madrid population (from xerophilous and pine forest) and the type locality population (from oak forest) (Table 2). Buccal tube length and body pores diameters are larger in the type locality population than in the west Madrid and the Caldera de Taburiente (Canary Islands) populations, where their values are similar. Body length and pharyngeal width are shorter in the type locality population than in the others (Table 2). Other species of tardigrades There is no other tardigrade species that occurs always in all the four localities where we have found Minibiotus gumersindoi n. sp. Echiniscus trisetosus Cuénot is present in three of the four localities (Bustarviejo valley, Cercedilla and Caldera de Taburiente). We present other tardigrade species found together with the new species in Table 4, where information is given on which species are new records for Madrid, Canary Islands and/or the Iberian Peninsula.