Integrative taxonomy reveals three new species of European Lepidocyrtus lignorum-group (Collembola, Entomobryidae) Author Mateos, Eduardo Departament de Biologia Evolutiva, Ecologia i Ciències Ambientals; Facultat de Biologia; Universitat de Barcelona; Avinguda Diagonal 643, 08028 - Barcelona; Spain. & Institut de Recerca de la Biodiversitat (IRBio), Universitat de Barcelona, Barcelona, Spain. Author Álvarez-Presas, Marta University of Bristol, School of Life Sciences, Bristol, United Kingdom. text Zootaxa 2022 2022-02-22 5100 4 451 481 journal article 20529 10.11646/zootaxa.5100.4.1 38ddf0a4-c35b-47a4-bf6a-5851ea91b0b4 1175-5326 6224598 74EEFDED-EEB9-46DF-83D5-2FB5693F920E Lepidocyrtus milagrosae Mateos sp. nov. Figs 24–40 , Tabs 1–2 Zoobank: urn:lsid:zoobank.org:act: 1AE4D1FD-A06D-491B-A9EA-47AB98D233B1 Type material. Holotype : male on slide (CRBA-90746), Laerma , Rhodes ( Greece ), 253 m above sea level , lat/long coordinates N36.146639 E27.920374 , on herbaceous vegetation and soil litter, hand collecting, 3.iv.2009 , leg. E. Mateos. Paratypes : 9 specimens without visible sexual plate on slides and 9 specimens preserved in absolute alcohol, same data as holotype . Holotype and paratype slide CRBA-90747 saved in the collection of the Centre de Recursos de Biodiversitat Animal , Faculty of Biology , University of Barcelona (http://www.crba.ub.edu); other paratypes kept in the E. Mateos’ collection (lot LP250 ) . Other material. 8 specimens preserved in absolute alcohol, Butterfly Valley , Petaloudes , Rhodes ( Greece ), 225 m above sea level , lat/long coordinates N36.337087 E28.062473 , on herbaceous vegetation (lot LP248 ) and soil litter (lot LP246 ), 3.iv.2009 , leg. E. Mateos. All material kept in the E. Mateos’ collection . Diagnosis. With dark blue pigment on the dorsal and ventral sides of Th.II to Abd.III, Ant.II–IV, and cx.I–III. Th.II slightly projecting over head. Ant.I–II, legs, ventral tube and posterior region of manubrium with scales. Apical bulb on Ant.IV absent. Labial chaetotaxy M 1 M 2 REL 1 L 2 , R slightly shortened. Dorsal cephalic and body macrochaetae formula A 0 [A 2a ]A 2 A 3 Pa 5 /00/0101+3. Abd.IV without chaeta s and with 4–6 lateral pseudopori on BP4. Unguiculus lanceolate and with serrated (or finely serrated) outer margin. Molecular diagnosis. This species includes all populations that cluster with CoxII and EF sequences of the individuals LP250-4 to LP250-7 ( Table 1 ), with significant support in an adequate molecular delimitation model. Etymology. The species is named after author’s wife Milagros (in apposition). Description . Holotype body length (without head nor furca) 1.2 mm , paratypes 1.0– 1.2 mm . Body colour pattern ( Fig. 24 ) with dark blue pigment on the dorsal and ventral sides of Th.II to Abd.III (including ventral tube), Ant.II–IV (with increasing colour intensity towards the distal part of each segment), and cx.I–III; densely black pigmented ocular areas. Mesothorax slightly projected over the head. FIGURE 24. Lepidocyrtus milagrosae sp. nov. : Habitus lateral (specimen in alcohol, without scales). Antenna with scales on dorsal region of Ant.I–II. Ratio antenna:cephalic diagonal = 1.4–1.6 (head diagonal measured from cervical edge to apex of mouth part); ratio Ant.I:II:III:IV as 1:1.8:1.8:2.8–3.2. Basis of Ant.I dorsally with three microchaetae arranged in triangle (Ant.I-organ); apex of Ant.I with a short curved S-chaeta in the membranous area of the ventral region. Ant.III organ composed of two subcilindrical and curved sensory rods. Ant.IV without apical bulb. 8+8 eyes; eyes A–F of equal size, eyes G and H a little bit smaller, ratio A/F and A/G = 1.3. Clypeus ( Fig. 25 ) with three prefrontal chaetae (1 pf0 and 2 pf1), facial area with four chaetae and three scales, and four lateral chaetae (2 L1 and 2 L2), all these chaetae ciliated. Prelabral and labral chaetae in typical number 4/554 ( Fig. 25 ), prelabral chaetae ciliated, first and second rows of labral chaetae smooth, apical row of labral chaetae branched ( Fig. 26 ); inverted U-shaped labral apical intrusion; four rounded labral papillae with three small pointed expansions ( Fig. 26 ). Maxillary palp outer lobe with smooth basal chaeta and apical appendage; sublobal plate with three smooth appendages and a smaller fourth one ( Fig. 27 ). Lateral process of outer labial papilla fingershape, slightly curved, tip not reaching apex of papilla ( Fig. 28 ). FIGURES 25–29. Lepidocyrtus milagrosae sp. nov. : 25, clypeal and labral chaetotaxy; 26, apical labral chaetae and papillae; 27, maxilary palp (right side); 28, outer labial papilla (right side); 29, labial and postlabial chaetotaxy (right side). Labial and postlabial chaetotaxy as in Fig. 29 ; with five smooth proximal chaetae at the base of labial palp; labial anterior row formed by five smooth chaetae (a1–a5); posterior row formed by ciliated chaetae with formula M 1 M 2 REL 1 L 2 ; chaeta R shorter, ratio M / R ≈ 2.3; postlabial chaetotaxy with all chaetae ciliated, row I (along ventral cephalic groove) with 4 chaetae. Dorsal cephalic macrochaetae formula A 0 A 2 A 3 Pa 5 , but also with pair of smaller supplementary macrochaetae A 2a between A 0 and A 2 , this chaeta A 2a can have two different morphologies depending on the specimens ( Fig. 30 ); maximum number of macrochaetae An on head 11+11 ( Fig. 30 ). Interocular chaetotaxy with s , t , p ciliated chaetae and 2–3 scales. Dorsal body macrochaetae formula 00/0101+3 (macrochaetae m3 on Abd.II , and Sm +B4, B5, B6 on Abd. IV). Dorsal chaetotaxy of Th.II –III and Abd.I as in Figs 31–33 . Th.II with 2 lateral S-chaetae ( al and ms ) and without macrochaetae in dorsal position. Th. III with a lateral sensillum ( al ) close to several ciliated chaetae. Abd.I with a lateral S-microchaeta ( ms ) external to a6 . Chaetotaxy of Abd. II–III as in Figs 34–35 . Abd.II chaeta ml present or absent depending on the specimens, chaeta p5p absent; macrochaetae m3 and m5 with equal socket diameter. Abd. III chaeta mi present or absent depending on the specimens, chaeta d3 absent, with S-chaetae as and ms, one specimen with a supernumerary associate ciliated chaeta between ll and a6 on Abd.III trichobothria a5 . All chaetae associated with the trichobothria on Abd.II –III acuminate and strongly ciliate. Chaetotaxy of Abd. IV as in Fig. 36 ; macrochaetae Sm, B 4, B5, B6, D3, E2, E3, E4, F1, F2, F3 broader and with broad socket; macrochaetae T6 , T7 , D2, De3, E1, E4p, Fe4, F3p shorter or longer but always thinner and with socket of minor diameter; macrochaeta F2 inserted above macrochaeta E3 ; the ratio of distances between macrochaetae Sm–B4 / B4–B6 is 0.6–0.8; the ratio of distances between macrochaetae B4–B5 / B5–B6 is 1.0–1.4; accessory chaeta s associated with trichobotrium T2 absent; chaetae a , D1 , m , pe and pi associated with trichobotria T2 and T4 acuminate and strongly ciliate; sens chaetotaxy composed of one anterior dorsomedial elongate S -chaetae, and short chaetae as and ps ; posterior margin with 5+5 smooth mesochaetae; lateral region with 4–7 small pseudopori on BP4 ( Fig. 37 ). Dorsal chaetotaxy of Abd.V with S-chaetae as , acc.p4 and acc.p5 ( Fig. 38 ). FIGURE 30. Lepidocyrtus milagrosae sp. nov. : Dorsal head chaetotaxy (left side). On the right side, the arrangement and morphology of the apical chaetae is drawn (note the different morphology that chaeta A2a may present). Broad circles––long ciliated macrochaetae, small circles––short ciliated macrochaetae. FIGURES 31–33. Lepidocyrtus milagrosae sp. nov. dorsal chaetotaxy (left side): 31, Th.II; 32, Th.III; 33, Abd.I. Circles––ciliated chaetae. FIGURES 34–35. Lepidocyrtus milagrosae sp. nov. dorsal chaetotaxy (left side): 34, Abd.II; 35, Abd.III. Broad circles––broad ciliated macrochaetae, small circles––thin ciliated macrochaetae. Legs with scales except in claws. V-shaped trochanteral organ formed by a maximum of 9 smooth straight chaetae ( Fig. 39 ). Unguis with basal pair of teeth at 48% from base of the inner edge, and with two inner unpaired teeth at 62% (the bigger) and 81% from base of the inner edge respectively; one external tooth and a pair of lateral teeth also present. Unguiculus lanceolate with smooth or finely serrated outer margin. Tibiotarsal tenent hair spatulate and smooth ( Fig. 40 ); ratio of tibiotarsal tenant hair / unguis inner edge ≈ 1.3; ratio of supra-empodial chaeta / unguiculus ≈ 1. Ventral tube with 6+6 ciliated chaetae on anterior side (4+4 proximal, 2+2 distal) and 10+10 weakly ciliated chaetae on posterior side; scales present only on anterior side; lateral flap with a maximum of 12 laterodistal chaetae (7 ciliated and 5 smooth). Manubrium with scales on anterior and posterior surfaces, with 2+2 ciliated apical chaetae on anterior side. The ratio manubrium:dens:mucro is 17:19:1. Manubrial plate with 2–3 inner chaetae and a maximum of 8 outer chaetae. Dental tubercle absent. Mucro with the two teeth of the same size, without spinelet on basal spine. Pseudopores distribution on dorsal and ventral positions as in Figs 23a,c . Ecology and distribution . All specimens were obtained by beating the herbaceous vegetation and sifting soil surface litter under holm oak trees. FIGURES 36–37. Lepidocyrtus milagrosae sp. nov. : 36, Abd.IV chaetotaxy (left side); 37, basal plate of Abd.IV. Broad black circles––broad ciliated macrochaetae, small black circles––thin ciliated macrochaetae, crossed circles––pseudoporus. FIGURES 38–40. Lepidocyrtus milagrosae sp. nov. : 38, Abd.V chaetotaxy (left side), broad black circles––long ciliated chatae, small black circles––short ciliated or smooth chaetae; 39, trochanteral organ; 40, third leg unguis and unguiculus with three different possibilities of unguiculus outer margin. Discussion . Morphological characters clearly assign Lepidocyrtus milagrosae sp. nov. to the Lepidocyrtus lignorum -group ( sensu Mateos 2011 ). By the characteristic body colour pattern L. milagrosae sp. nov. clearly differs from all the other species of the L. lignorum -group except L. labyrinthi . These two species differ by the morphology of labral papillae (with three pointed expansions in the new species and only one in L. labyrinthi ) and the presence of pseudopores on BP 4 in the new species ( Table 2 ). Other characters (not included in Table 2 ) differentiating L. labyrinthi and the new species are the absence of chaetae a5 , m5, and p6 on Abd.I, and the presence of chaetae p5p on Abd.II and d3 on Abd.III in L. labyrinthi ( Baquero et al . 2021 ) . By having body partially blue pigmented and lanceolate unguiculus L. milagrosae sp. nov. is close to species L. instratus , L. traseri , L. fuscocephalus sp. nov. , and L. semicoloratus sp. nov. Of these, the new species differ by the colour pattern and the presence of pseudopores on BP4. Other differences between all species included in the group are summarized in Table 2 .