Zoosyst. Evol. 100 (2) 2024, 469-482 | DOI 10.3897/zse.100.119153 > PENSUFT. Gee Ee BERLIN A new species of genus Urophonius Pocock, 1893 (Scorpiones, Bothriuridae), from Andean Mauline Chilean forests, with a phylogenetic re-analysis of the genus Andrés A. Ojanguren-A ffilastro!, Fermin M. Alfaro***, Martin J. Ramirez’, Bernardino Camousseigt-Montolivo°, Jaime Pizarro-Araya 2,4,6,7 1 Division Aracnologia, Museo Argentino de Ciencias Naturales “Bernardino Rivadavia” (CONICET), Avenida Angel Gallardo 470, 1405 DJR, Buenos Aires, Argentina 2 Laboratorio de Entomologia Ecolégica (LEULS), Departamento de Biologia, Facultad de Ciencias, Universidad de La Serena, Casilla 554, La Serena, Chile Instituto de Ecologia y Biodiversidad (IEB), Santiago, Chile N DO Oo ff W https://zoobank. org/1A6B3F 2D-ADB7-412B-A2A D-BB7A9BC35757 Corresponding author: Jaime Pizarro-Araya (japizarro@userena.cl) Programa de Doctorado en Biologia y Ecologia Aplicada, Universidad Catolica del Norte, Universidad de La Serena, La Serena, Chile Grupo de Artrépodos, Sistema Integrado de Monitoreo y Evaluacion de Ecosistemas Forestales Nativos (SIMEF), La Serena, Chile Environment & Permitting - HSEQ, Enel Green Power & Thermal Generation, Roger de Flor 2725, Torre 1, Piso 1, Las Condes, Santiago, Chile Programa de Doctorado en Conservacion y Gestion de la Biodiversidad, Facultad de Ciencias, Universidad Santo Tomas, Santiago, Chile Academic editor: Danilo Harms # Received 19 January 2024 # Accepted 28 March 2024 Published 7 May 2024 Abstract Urophonius trewanke sp. nov. is described from the Mauline Andean woods of northern Chilean Patagonia. This species belongs to the granulatus species group, which includes the most basal species within the genus. This species is only active in summer as in all species of its group. We performed a phylogenetic analysis of the genus Urophonius based on morphological characters to establish the position and relationships of the new species in the genus. Key Words Bothriuridae, Chile, Mauline woods, new species, phylogeny Introduction The scorpion genus Urophonius Pocock, 1893 comprises small burrowing species from southern South America. In the last two decades, there has been a remarkable increase in the knowledge of this genus; in this period, the num- ber of described species has almost doubled (Acosta 2003; Ojanguren-Affilastro and Cheli 2009; Ojanguren-A ffilas- tro et al. 2010, 2011, 2020), the first phylogenetic analysis of the genus has been performed (Ojanguren-A ffilastro et al. 2020) and, more recently, the historical time frame of its evolution and diversification has been unveiled using tran- scriptomes and UCEs (Ojanguren-A ffilastro et al. 2023). Urophonius is remarkable amongst all known scor- pion genera from temperate and cold areas because of its adaptations to low temperatures (Maury 1969, 1973; Ojanguren-A ffilastro et al. 2020; Garcia et al. 2021). This genus reaches the southernmost and colder part of South America (Maury 1979; Ojanguren-A ffilastro et al. 2020) and most Urophonius species exhibit surface activity in winter, contrary to other species from the region (and temperate areas of the world), which are active during the warmer period of the year (Pizarro-Araya et al. 2011; Ojanguren-A ffilastro and Kovarik 2013). The first morphological phylogeny of the genus (Oyan- guren-A ffilastro et al. 2020) suggested a single and relatively Copyright Ojanguren-Affilastro, A.A. et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 470 Ojanguren-Affilastro, A.A. et al.: New Urophonius from the Andean Mauline forests early origin of winter activity in Urophonius. A posterior dated phylogeny, based on diverse phylogenomic datasets (Ojanguren-A ffilastro et al. 2023), supported this early or- igin of the winter activity period and dated it between 68 and 42 million years ago (MYA), before the most important uplift of the Andes (Ghosh et al. 2006; Garzione et al. 2008). On the other hand, some species of the genus still re- tain the summer activity period, which is common to most species of the family and the order (Ojanguren-A ffilastro and Kovarik 2013). These summer Urophonius species are grouped in the granulatus species group and occur exclusively in cold areas of southern South America, both in steppes and temperate woods. In a recent comprehensive study of arthropods con- ducted at the “Fundo La Escuadra” (Figs 1a, 2), a small well-preserved area located within the Cipreses Riv- er Basin, a tributary of the Maule River Basin in the Maule Region of Chile, our research team made notable discoveries. The upper Maule River Basin is partially isolated by the Andes and has proven to harbour sever- al endemic species of arthropods and even vertebrates (Corbalan et al. 2010; Correa et al. 2018, 2020; Ojan- guren-Affilastro et al. 2020). This study allowed us to identify and collect numerous species of arachnids and insects not previously documented. In particular, amongst these newly-discovered taxa, we found a hith- erto unknown species of scorpion that belongs to the genus Urophonius Pocock, 1893. This research, rooted in the rich and highly endemic biodiversity of the area, contributes significantly to our understanding of the local arthropod fauna. The identification and documen- tation of a new scorpion species underlines the ecolog- ical importance of the ecosystem preserved at “Fun- do La Escuadra”. The findings of this study not only expand our knowledge of regional arthropod diversity, but also emphasise the need for further research and conservation efforts in this ecologically important area. In this contribution, we describe Urophonius trewan- ke sp. nov. (Fig. 1b) from the Maule Valley in the up- per Maule River Basin (Fig. 2b). Urophonius trewanke sp. nov. is the second known endemic Urophonius from this area, the other species being Urophonius pehuenche Ojanguren-A ffilastro & Pizarro-Araya, 2020, which, con- trary to the new species, is only active in winter (Ojan- guren-Affilastro et al. 2020). We also perform a phylo- genetic analysis, based in morphological characters to clearly stablish the phylogenetic position of the new spe- cies in the genus. Methods Cladistic analysis Taxa. The matrix in the cladistics analysis comprises a total of 21 species, 17 species of Urophonius and four outgroups. We used the same species as Ojanguren-Af- filastro et al. (2020), including all known species of zse.pensoft.net Urophonius, together with the new species herein de- scribed: U. trewanke sp. nov. Outgroups belonging to four other genera in the family Bothriuridae include: one species of the Australian bothriurid Cercopho- nius, which, according to all previous phylogenetic analyses, is the sister genus of Urophonius (Prendini 2000, 2003; Ceccarelli et al. 2016; Ojanguren-A ffilas- tro et al. 2020, 2023; Santibafiez-Lopez et al. 2023); one species of Phoniocercus Pocock, 1893, which has also been suggested as closely related to Urophonius (Acosta 1988); one species of Thestylus Simon, 1880, which, according to previous phylogenetic analysis, is the most basal split in the American bothriurids (Pren- dini 2000, 2003; Ceccarelli et al. 2016); and one spe- cies of Centromachetes Lonnberg, 1897, which also shares several morphological characters with Uropho- nius (Ojanguren-Affilastro and Kovarik 2013). The tree was rooted on Thestylus aurantiurus Yamaguti & Pinto-da-Rocha, 2003, based on previous evidence for the relationships amongst bothriurid genera (Prendini 2000, 2003). Materials Specimens examined are deposited in the following collections: Museo Argentino de Ciencias Naturales “Bernardino Rivadavia,’ Buenos Aires, Argentina (MACN-Ar, Martin J. Ramirez); Museo Nacional de Historia Natural, Santiago, Chile (MNHN, Mario Elgueta Donoso); Museo de Zoologia de la Universidad de Concepcién (MZUC, Laura Tavera Martinez); Laboratorio de Entomologia Ecologica, Universidad de La Serena, Chile (LEULS, Jaime Pizarro-Araya). A table with exemplars and locality data used for the analyses are presented as Suppl. material 1. Characters We used a matrix, based in 114 morphological charac- ters. The complete list of characters and the matrix are available as supplementary material online, as Suppl. ma- terials 2, 3, respectively. We used the same matrix as in Ojanguren-Affilastro et al. (2020) with the sole addition of the new species herein described. Analyses Analyses were made with TNT 1.5 (Goloboff et al. 2008), using parsimony under equal weights and implied weights, exploring the sensitivity of the results to a range of values of the concavity constant & from 1 to 99. Branch support was estimated with 1000 pseudoreplicates of symmetric resampling. Tree searches were made with 100 random addition sequences (RAS) followed by tree bisection-reconnection (TBR) branch swapping; since all Zoosyst. Evol. 100 (2) 2024, 469-482 the RAS+TBR replicates reached the same result, it is likely that optimal trees were found. Systematics All new material reported here was collected by the au- thors; most specimens were collected at night by UV de- tection. Some specimens were also collected in daytime under logs or stones. Measurements, taken using an ocular micrometer, are recorded in mm. Descriptive terminolo- gy follows Mattoni and Acosta (2005) for hemispermato- phores; Vachon (1973) for trichobothria; Francke (1977) for metasomal carinae abbreviated as follows: DL: dorso- lateral; LIM: lateral inframedian; LSM: lateral suprame- dian; LM: lateral median; VSM: ventral submedian; VL: ventrolateral; VM: ventromedian; and Prendini (2000) for pedipalp carinae, abbreviated as follows: DI: dorsal internal; DE: dorsal external; VI: ventral internal; VE: ventral external; D: digital; E: external; IM: internomedi- an; EM: externomedian; V: ventral; VM: ventral median; DM: dorsal marginal; DS: dorsal secondary. Illustrations were produced with a Leica M165C stereomicroscope and a camera lucida. Digital images of pigmentation pat- tern and habitus were taken under visible light, images of external morphology under UV light, using a digital camera (Leica DFC290 or Nikon DS-Fil) attached to a stereomicroscope (Leica M165C or Nikon SMZ1500) and the focal planes combined with Helicon Focus 3.10.3 (http://helicon.com.usa/heliconfocus/). Point locality re- cords were georeferenced in the field with portable Glob- al Positioning System devices (Garmin® Etrex Vista and Etrex Vista C). The distribution map was generated using https://www.simplemappr.net/. Results Cladistic analyses The analysis under equal and implied weights resulted in highly concordant trees. All the analyses under implied weights with concavity constant below 20 produced the resolution of Fig. 3. The support values were calculated under implied weights with k= 15. In our phylogenetic tree (Fig. 3), the genus Uropho- nius appears as monophyletic and Cercophonius appears as the sister genus of Urophonius, as in previous phylo- genetic analyses (Prendini 2000; Ojanguren-A ffilastro et al. 2020). We recovered two major clades confirming the results of Acosta (1988) and Ojanguren-Affilastro et al. (2020). One of these corresponds to the concept of granulatus Species group (Fig. 3) and includes all species with sum- mer activity period and the new species herein described. In our analyses, Urophonius trewanke sp. nov. groups with Urophonius tregualemuensis Cekalovic, 1981, as expected by their external similarities and environmen- 471 tal proximity, both inhabiting southern Chilean woods. Species from the Patagonian steppe of the granulatus group, Urophonius granulatus Pocock, 1898, Uropho- nius somuncura Acosta, 2003 and Urophonius araucano Ojanguren-Affilastro & Pizarro-Araya, 2020 form a sep- arate clade. Urophonius pizarroi Ojanguren-Affilastro, Ochoa, Mattoni & Prendini, 2010, fits in the granulatus group, but it is not part of either clade. On the other hand, we recovered all species with win- ter activity as another monophyletic group (Fig. 3), with two internal clades corresponding to exochus and brachy- centrus groups (Acosta 1988; Ojanguren-Affilastro et al. 2020). Urophonius mondacai Ojanguren-A ffilastro, Pizarro-Araya & Prendini, 2011 appears as basal to the species with winter activity; however, its actual activity period is in debate. The original records of this species are from spring, but it has recently been collected also in winter by the authors, raising more doubts about the position of this enigmatic species. Systematics Urophonius trewanke sp. nov. https://zoobank. org/50D20A 04-1 E7A-47E1-9EF3-3C60F4BA 8922 Figs 1-8; Table 1 Type material. Chile, Maule Region (VII), Maule Valley, Fundo La Escuadra: Holotype 4 (MNHN 8411), Bocato- ma-Ojos de Agua (35°46'06.1"S, 70°47'44.4"W), 1009 m a.s.1.; 14-17/X/2022, Pizarro-Araya, Alfaro & Calderon coll. Paratipes: 2 4, same data as holotype (MACN); La- guna Invernada (35°43'16.1"S, 70°47'04.9"W), 1260 m a.s.1.; 14-17/X/2022, Pizarro-Araya, Alfaro & Calderon coll. 2 9, 10 ¢ (LEULS); 2 2, 2 4 (MACN). Woods of Quillaja saponaria and Cryptocarya alba (35°46'04.1"S, 70°47'45.6"W), 1020 m as.l.; 10/XII/2023, Pizarro- Araya, Alfaro & Calderon coll. 3 2 (LEULS); 3 9, 6 4 (MACN). Bocatoma-Ojos de Agua (35°46'06.1"S, 70°47'44.4"W), 1009 maz.s.1.; 9/XTI/2023, Pizarro-Araya, Alfaro & Calderén coll. 4 9, 2 4, 1 juvenile (LEULS), 2 2,3 d (MACN). Etymology. The specific epithet “trewanke” is a noun in apposition meaning scorpion in Mapungudun, the lan- guage of the Mapuche people, the original inhabitants from most parts of southern and central Chile. Diagnosis. Urophonius trewanke sp. nov. is most closely related to U. tregualemuensis from south-central Chile (Fig. 2a). Both species can be easily separated by their pigment pattern; in U. trewanke sp. nov. the dor- so-submedian spots of tergites are poorly developed, be- ing reduced to small triangles in the posterior half of the segment (Figs 4a, 5a), whereas in U. tregualemuensis, these spots are much more developed, occupying almost the whole median part of the segment (Fig. 5b). Both species can also be separated by the shape of pedipalp chela, which is stouter in U. trewanke sp. nov. (Fig. 6b, f). Pedipalp chela length/height ratio varies from zse.pensoft.net 472 Ojanguren-Affilastro, A.A. et al.: New Urophonius from the Andean Mauline forests sare: Coe rae ee i DES ON oli, eh rk SPOS ae Figure 1. a. Habitat of Urophonius trewanke sp. nov., woods at Fundo La Escuadra, Maule Valley, Maule Region, Chile; b. Couple of Urophonius trewanke sp. nov. during courtship in its natural environment; ¢. Urophonius trewanke sp. nov. male, living specimen. 3.15-3.38 (n = 11, mean = 3.29) in U. trewanke sp. nov. males and between 3.96 and 4.24 (n = 12, mean = 4.09) in U. tregualemuensis males; it varies between 3.31 and 3.61 (n= 3, mean = 3.45) in U. trewanke sp. nov. females and between 4.06 and 4.33 (n = 10, mean = 4.19) in U. tregualemuensis females; length/width ratio varies be- tween 3.35 and 3.70 (n= 11, mean = 3.54) in U. trewanke sp. nov. males and between 4.13 and 4.24 (n= 12, mean = 4.09) in U. tregualemuensis males; and varies between 3.70 and 3.93 (n= 3, mean = 3.80) in U. trewanke sp. nov. females and between 4.48 and 4.82 (n= 10, mean = 4.63) in U. tregualemuensis females. They can also be separated by some details of the hemispermatophore; in U. tregualemuensis, the bifid lobe of the internal lobe 1s connected to the superior concavity zse.pensoft.net of the basal portion by a thick carina (Fig. 8b), that is very subtle in U. trewanke sp. nov. (Fig. 8a). Additionally, the distal margin of capsular concavity of the basal lobe is arranged more distally in U. trewanke sp. nov. (Fig. 8c, d) than in U. tregualemuensis (Fig. 8e, f). There are also some differences in the development of the ventral carinae of metasomal segments I and IT, which are Clearly more developed in U. tregualemuensis (Fig. 7d, e) than in U. trewanke sp. nov. (Fig. 7a, b), particu- larly in males, in which these carinae are barely visible in U. trewanke sp. nov. (Fig. 7a) and well developed in U. tregualemuensis (Fig. 7d). Description. Based on the holotype @ (MNHN) and the paratypes 9 (LEULS, MACN-Ar). Zoosyst. Evol. 100 (2) 2024, 469-482 a Se * Fy Saas @ Urophonius trewanke n. sp. | +> Urophonius tregualemuensis | ¢ gd 70°40'48"W is, 5 35°43'48"S 35°43'48"S Aa. Hydroelecttic Plant, © /Losipreses ~ Maule River. LaMina E ° i : & Tang | “La Escuadra” boundaries @ Urophonius trewanke n. 5| [4 © Cities or towns 70°58'48"W Maule Rive! ~ 35°52'48"S 35°52'48"S 70°49'48"W 70°40'48"W Figure 2. a. Map of central Chile with the distribution of Uro- Phonius trewanke sp. nov. and its closest species, Urophonius tregualemuensis; b. Map of the study area, marking the collec- tion sites, the Cipreses River and the Maule River. Total length: 30-41 mm in @ (n = 12; mean = 34.7); 34.541 mm in 9 (n= 4; mean = 38.88). Colour: Base colour yellowish, with dark brown spots (Figs 1b, c, 4). Chelicerae with reticulate pigmentation on dorsal and retrolateral surface of manus, densely pig- mented on the retrolateral margins of the fingers. Cara- pace, densely pigmented (Fig. 5a); anterior margin pig- mented, with two broad, dark lateral stripes, extending from lateral sides of the anterior margin to the ocular tu- bercle and the anterior part of posterior longitudinal sul- cus, with two other lateral dark stripes placed more pos- teriorly and reaching the lateral margins; median ocular tubercle and area around lateral ocelli dark brown; with two posterolateral triangular dark spots covering most of latero-posterior margin, leaving a median unpigmented area in the postocular furrow. Tergites I-VI each with four dark spots (Fig. 5a), two external-lateral forming a stripe along the segment and two submedian-subtriangular in the posterior half of the segment; lateral and median spots can be connected by pigment in the posterior margin, me- 473 dian area of the segment always unpigmented; tergite VII with two postero-lateral spots on each side, posterior to dorso-median and dorso-lateral carina, respectively. Ster- num, genital opercula, pectines and pleura unpigmented. Sternites: sternites II], [IV and V unpigmented medially (Fig. 5c), pigmented in the lateral margins; sternites VI and VII, lateral margins pigmented, posterior margin with faint posterior VL spots and a VM posterior spot that can continue in a thin median stripe, particularly in segment VII. Metasomal segments I-III: dorsal surface with two triangular posterior spots and two anterior small spots, with a thin stripe over the DL carina, with faint paired median spots that can be absent in some specimens; lat- eral surfaces densely pigmented between LM and LIM carinae; ventral surface: with VL stripes well marked, ex- tending the entire length of the segment, slightly thicker posteriorly, without VSM stripes, VM stripe thin and ex- tending the entire length of the segment, not connecting with VL stripes; segment IV similar to III, but the dorsal spots are elongated; segment V clearly darker than the rest of the segments, with a dark reddish base colour, with faint dorsal submedian stripes; with a lateral stripe along the LM carina, connecting with the VL stripe in the sec- ond half of the segment, postero-lateral margins densely pigmented, ventrally similar to remaining segments, but with VL stripes connected to lateral stripes. Telson, gen- eral colour dark reddish-brown, as metasomal segment V, dorsal gland of males barely paler than the rest of the vesicle; aculeus dark brown. Pedipalps, trochanter dor- sally pigmented; femur with DI, DE and VE wide stripes across the whole segment, fusing in the articulation with patella. Patella, with a DI reticular stripe extending the entire length of the segment, but with an unpigmented median area, with a DE thin stripe in the proximal third of the segment and a retrolateral median stripe extending the entire length of the segment. Chela with seven dark stripes which seem to correspond to DI, DM, DS, D, E, V and VM carinae; area near articulation of fingers and fingers, pigmented. Legs: coxae unpigmented; trochanter with a median retrolateral spot; femur, retrolateral mar- gin with anterior and posterior elongated spots; patella with retrolateral and dorsal stripes; tibia with dorsal and ventral spots; basitarsi with ventral and dorsal spots near articulation with tibia; telotarsi with a dorsal faint spot near articulation with basitarsi. Carapace: lateral surfaces granular (32), medial- ly smooth (@) or slightly granular (<). Anterior margin straight. Anterior longitudinal sulcus shallow; interocu- lar sulcus weakly developed; posterior longitudinal and lateral sulci well developed. Median ocular tubercle not very pronounced, median ocelli large, ca. one diameter apart; with one macroseta behind each eye and one mi- croseta in front of each eye. Three pairs of small lateral ocelli on each side of carapace, posterior ocellus slightly smaller than the rest of the ocelli; anterior and median ocelli almost in the same horizontal axis, posterior ocel- lus situated clearly dorsal to others; lateral ocelli pattern type 3A (Loria and Prendini 2014). zse.pensoft.net 474 Ojanguren-Affilastro, A.A. et al.: New Urophonius from the Andean Mauline forests Thestylus aurantiurus Be Phoniocercus sp. 100 Centromacehetes sp. Lf Cercophonius sulcatus Genus Urophonius ——~> 100 Urophonius pizarroi 22 [rls 23 + Urophonius trewanke n. sp. Urophonius tregualemuensis Urophonius araucano a ba Urophonius somuncura granulatus group Urophonius granulatus - Urophonius mondacai Urophonius exochus a Urophonius mahuidensis . |e@xochus grou Urophonius martinezi ales Se sphonius eugenicus : Species Urophonius tumbensis brachycentrus| with Urophonius pehuenche oe : 64 | Lee oot transandinus group winter 70 activity Urophonius iheringi ‘ae 2 Sec orus achalensis Urophonius brachycentrus Figure 3. Phylogeny of the genus Urophonius Pocock, 1893 estimated with morphological characters under implied weights (k = 15), with Jackknifing frequencies indicated inside branches. Pedipalps: Femur with DI, DE and VI carinae granu- lar, extending the entire length of segment (Fig. 6h); with some sparse coarse granules in the anterior margin of the segment; with one macroseta related to trichobothria d and 7; trichobothrium e situated distal to dorsal macrose- ta M1 (Fig. 6h). Patella with smooth tegument and with- out distinct carinae (Fig. 61). Chela manus robust, more so in @, length/width ratio varying from 3.15—3.38 in @ (n = 11, mean = 3.29) and from 3.31—3.61 in 2 (n =3 mean = 3.45); length/height ratio varies from 3.35—3.70 in S (n= 11, mean = 3.54) and from 3.70-3.93 in 9 (n =3, mean = 3.80); acarinate (Fig. 6a—e, g), prolateral surface with a pronounced, subtriangular projection and a shallow depression near articulation of movable finger (Fig. 6b), with a group of granules near the base of the movable fin- ger (4); all of them absent in 9 (Fig. 6c, g); fingers elon- gated, median denticle row medially uneven in the basal quarter of its length, with five pairs of accessory granules. Pectines: Tooth count: 15-17 in 3 (n = 12, medi- an = 16) and 14-15 in 9 (n=4, mean = 15). Legs: Surfaces smooth in 9, granular in <4. Basitarsi each with two well developed, equal length, pedal spurs. Telotarsi elongated, shallow, each with well-developed ventromedian row of hyaline setae and paired rows of ventrosubmedian spiniform setae with the following counts on each telotarsus: I: 1/1, II: 2/2, Ill: 5—6, IV: 6-6/6—7. The only pair of spines of telotarsus I and the first pair of spines of telotarsus II are less sclerotised than the remaining spines, the rest are well sclerotised. Ungues slightly curved, equal in length. Tergites: Surfaces, I-VI: anterior area smooth, poste- rior and lateral margins finely granular; more so in 3; VII with sparse, coarse granules in posterolateral margins, with paired dorso-submedian carinae in posterior third and paired dorso-lateral carinae in posterior two-thirds of the segment. zse.pensoft.net Sternites Surfaces, II—VI smooth, with small ellipti- cal spiracles; VII, surface sparsely granular, more so in S:; in 2 with two VM and two VL barely visible carinae in posterior third of the segment, not conspicuous in 3 (Fig. 7a, b). Table 1. measurements in mm of the holotype male (MNHN) and a female paratype (MACN) of Urophonius trewanke sp. nov. Urophonius trewanke sp. nov. Holotype ¢ Paratype ° Total length S205 38.72 Carapace, length Shall an oul Carapace, anterior width 2.58 3339 Carapace, posterior width 4.12 Bey Mesosoma, total length 8.88 9.69 Metasoma, total length 19.94 21:02 Metasomal segment I, length 1.94 2.58 Metasomal segment |, width 2.42 3:23 Metasomal segment I, height 2.02 2.58 Metasomal segment Il, length 2.34 2.82 Metasomal segment Il, width 2.18 2.91 Metasomal segment Il, height 1.85 2.50 Metasomal segment Ill, length 2.58 3.47 Metasomal segment Ill, width 2.15 DFA Metasomal segment Ill, height 1.85 2.42 Metasomal segment IV, length 3.07 4.04 Metasomal segment IV, width 2.02 2.58 Metasomal segment IV, height 1.85 2.42 Metasomal segment V, length 4.68 5.41 Metasomal segment V, width 210 2.83 Metasomal segment V, height ey a 2.42 Telson, length 5.33 5.70 Vesicle, width leva: 2.26 Vesicle, height 1-53 1.82 Chela, length 6.06 7.62 Chela, width 1.85 1.94 Chela, height 1.64 2:10 Zoosyst. Evol. 100 (2) 2024, 469-482 475 Figure 4. Urophonius trewanke sp. nov., a, b. Holotype 4; ¢, d. Paratype 9; a, c. Dorsal aspect; b, d. Ventral aspect. Scale bar: 10 mm. zse.pensoft.net 476 Ojanguren-Affilastro, A.A. et al.: New Urophonius from the Andean Mauline forests Figure 5. Pigment pattern. a, e. Urophonius trewanke sp. nov. a. Carapace and tergites; ec. Sternites; b, d. Urophonius tregualem- uensis Cekalovic, 1981; b. Carapace and tergites; d. Sternites. Metasoma: Metasomal segment I, dorsal surface finely granular; DL carinae granular, extending the entire length of segment with anterior and posterior granules more developed than the rest; dorso-lateral margins granular, LSM carinae represented by some tiny granules in the posterior part of the segment, LM carina with an anterior blunt small keel and a granular part extending the pos- terior two-thirds of the segment; LIM carinae granular, restricted to the posterior half of the segment, with one macroseta; ventral surface smooth, VL carinae extending the entire length of the segment, granular in 9, as an ele- vation of the tegument in 3, VSM carinae well developed and granular in 9, barely visible in @, with two pairs of VSM macrosetae and three pairs of VL macrosetae (Fig. 7a, b). Metasomal segment II, similar to I, but with less developed carinae, being the ventral carinae barely vis- ible in 4; with a DL, a LM and a LIM macroseata and with three pairs of VSM and VL macrosetae. Metasomal segment HI similar to segment II, but with less developed carinae, without ventral carinae, LIM carina restricted to posterior third of the segment; metasomal segment IV elongated, LIM carina absent, the rest similar to segment III. Segment V elongated (Fig. 7d, e); dorsal and lateral margins smooth; DL carinae restricted to some well-de- veloped granules in the anterior third of the segment, with zse.pensoft.net two dorsal macrosetae and four lateral macrosetae; VL carinae granular, extending the posterior two-thirds of the segment, with five pairs of VL macrosetae, being the pos- terior pair in the posterior margin; ventral surface dense- ly granular in the posterior two-thirds of the segment, so that VSM and VM carinae are not conspicuous between the granulation (Fig. 7c, f), with four pairs of VSM mac- rosetae, being the posterior pair in the posterior margin. Telson: Vesicle, shallow, more lobular in °; ventral sur- face with medium sized granules in 9, less granular in 3; dorsal surface smooth, with (<') or without (@ ) an elliptical median well-developed depression corresponding to the telson gland. Aculeus short, shallowly curved (Fig. 7g, h). Hemispermatophore: Basal portion well developed. Distal lamina well developed, ca. 30% shorter than basal portion; distal crest almost straight, orientated almost in same direction to the posterior margin of the DL; frontal crest (distal posterior flexure) present; internal lobe with two well-developed denticles, not connected with the dis- tal lamina (Fig. 8a), external denticle ca. twice larger than internal denticle. Lobe region poorly developed (Fig. 8c, d); basal lobe well developed, barely protruding, with a flat internal laminar extension, with an internal concavi- ty; genital plug poorly developed, barely exceeding the capsular concavity. Zoosyst. Evol. 100 (2) 2024, 469-482 Figure 6. Left pedipalp. a-e, g—i. Urophonius trewanke sp. nov. a—e. Chela, 4; a. Dorsal aspect; b. Prolateral aspect; ¢. Ven- tro-prolateral aspect; d. Ventral aspect; e. Retrolateral aspect; g. Chela 2, prolateral aspect; h. Femur, @, dorsal aspect; i. Patella, &, retrolateral aspect; f. Urophonius tregualemuensis Cekalovic, 1981. Left pedipalp chela, <, prolateral aspect. Scale bars: 1 mm. zse.pensoft.net Ojanguren-Affilastro, A.A. et al.: New Urophonius from the Andean Mauline forests Figure 7. a—c, f-h. Urophonius trewanke sp. nov., a, b. Sternite V and metasomal segments I and II, ventral aspect a. 3 and b. 9: c. metasomal segment V, <', ventral aspect; f. Metasomal segment V, °, ventral aspect; g. Telson, 4, lateral aspect; h. Telson, &, lateral aspect; d, e. Urophonius tregualemuensis Cekalovic, 1981, sternite V and metasomal segments I and II, ventral aspect; d. 3; e. 2. Scale bars: 1 mm. zse.pensoft.net Zoosyst. Evol. 100 (2) 2024, 469-482 Figure 8. a, c, d, Urophonius trewanke sp. nov., a. Left hemispermatophore, external aspect; ce. Left hemispermatophore, lobe region, internal aspect; d. Right hemispermatophore, lobe region, internal aspect; b, e, f, Urophonius tregualemuensis Cekalovic, 1981: b. Left hemispermatophore, external aspect; e. left hemispermatophore, lobe region, internal aspect; f. Right hemispermato- phore, lobe region, internal aspect. Scale bars: 1 mm. Distribution. This species has only been collected in area is placed in the Maule Valley, in the Maule Region, its type locality, in the small preserved area of “Fundo Chile, close to the Pehuenche international pass which La Escuadra”, 35°46'06.1"S, 70°47'44.4"W (Fig. 1). This connects Chile with Argentina (Fig. 2). zse.pensoft.net 480 Ojanguren-Affilastro, A.A. et al.: New Urophonius from the Andean Mauline forests Ecology. The area where Urophonius trewanke sp. nov. has been collected is located within the landscape of the “Estepa de los Andes Maulinos” (Mauline Andean Steppe) Botanical Formation. This distinctive formation represents the southernmost extension of the high Ande- an steppes. From this point, towards the south, a change in ecological conditions is perceived, characterised by an increase in precipitation and snowfall. This transition marks a natural limit for the distribution of numerous southern and boreal species (Gajardo 1993). The ecological matrix of this region 1s made up of var- ious types of vegetation, which has defined its heteroge- neity. Dry forests intersect with shrub steppes, creating a plant mosaic. Notable vegetation components include spe- cies such as Chuquiraga oppositifolia D.Don, Gochnatia foliorosa D.Don and Proustia cuneifolia D.Don (Aster- aceae), each of which plays a role in shaping the unique habitat of Urophonius trewanke sp. nov. (Fig. 1a). In addition, the landscape presents different herbaceous steppes, where species such as Acaena alpina Poepp ex. Walp. (Rosaceae) and Festuca acanthophylla Desv. (Po- aceae) contribute to the overall floral composition. In the midst of this botanical diversity, the presence of the Cordil- lera cypress (Austrocedrus chilensis (D.Don) Pic-Serm. & Bizzarri.) stands out, which adds to the ecological tapestry with its characteristic shape and contributes to the general microhabitat where Urophonius trewanke sp. nov. occurs. This environment, with its varied types of vegetation and the inclusion of notable species, such as the Cypress of the mountain range, highlights the ecological importance of the “Estepa de los Andes Maulinos” Botanical Forma- tion as a unique and valuable habitat for the diversity of arachnids, providing crucial information on the ecology and habitat preferences of Urophonius trewanke sp. nov. The Maule Valley and its associated area along the Maule River in the Pehuenche Andean Pass, seems to constitute an area of endemism for the epigean fauna, separated from surrounding valleys by transverse moun- tain chains. This area is already known to harbour an en- demic and highly restricted species of Anuran, Alsodes pehuenche Cei, 1976 (Corbalan et al. 2010; Correa et al. 2018, 2020). In nearby localities outside the Maule Valley, but with similar habitats, U. trewanke sp. nov. is replaced by U. tregualemuensis, another species of the granulatus group, which occupies similar environments and niches as U. trewanke sp. nov., but has a wider distribution in south central Chile (Fig. 2a) (Ojanguren-Affilastro et al. 2011). Urophonius trewanke sp. nov. has a spring/summer activ- ity period, as in all species within the granulatus species group, therefore retaining what we consider the ancestral condition of the genus. In winter, in nearby areas of the Pe- huenche Valley, the only active species of Urophonius (and of the whole order) is U. pehuenche, also endemic from the Pehuenche Valley (Ojanguren-A ffilastro et al. 2020). Urophonius trewanke sp. nov. has been collected in sympatry with an undescribed species of Brachistosternus, zse.pensoft.net which also seems to be endemic of the Pehuenche Pass (Ojanguren-A ffilastro et al. in prep.). Discussion The description of U. trewanke sp. nov., a rare species belonging to the granulatus group from southern Chile, supports our previous hypothesis that the few species of Urophonius with a summer activity period, are re- stricted to central and southern Chile and to the colder areas of southern South America, whereas the species with a winter activity period are more widely distribut- ed (Ojanguren-A ffilastro et al. 2020, 2023). Our analyses included all known species of the genus and most of the characters used in all recent morphological phylogenies in the family; the lack of major changes in the general topology respect to our previous phylogeny in the genus (Ojanguren-Affilastro et al. 2020) despite the inclusion of a new species to our analysis, provides more support to our previous hypothesis about the relationships inside the genus. Urophonius trewanke sp. nov. has been collected in “Fundo La Escuadra’, meaning La Escuadra farm or ranch, a small preserved area currently under manage- ment by ENEL (“Empresa Nacional de Energia Eléc- trica” or National Electric Energy Company). This area has been part of the “Cipreses” operational system of the Chilean Electric National System since 1955 and _ has, therefore, had highly restricted access for the last seventy years. This inaccessibility has resulted in an unintended, but yet remarkable, degree of preservation. This area 1s placed in the partially isolated Maule Valley which has proved to harbour an exceptional number of endemics, but lacks any kind of formal protection. We hope that the description of the actual diversity of this area could shed light on the need to preserve this important biological resource and lead to future preservation of the endemic species of the Maule Valley. Acknowledgements Our special thanks to the staff of the Los Cipreses hydro- electric power plant, especially to Christian Cartes for his help in logistics (La Escuadra, ENEL). Project funded by Environment & Permitting - HSEQ, Enel Green Power & Thermal Generation (ENEL). J.P-A thanks the Academ- ic Excellence Scholarship (B134) from the Academic Vice-Rector’s Office, Research and Postgraduate Stud- ies of the Universidad Santo Tomas, Santiago, Chile and ANID doctoral fellowship 2024. F.M.A. thanks the ANID doctoral fellowship 2023-21230592. 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Ramirez, Bernardino Camoussel- gt-Montolivo, Jaime Pizarro-Araya Data type: docx Copyright notice: This dataset is made available under the Open Database License (http://opendatacommons. org/licenses/odbl/1.0/). The Open Database License (ODbL) 1s a license agreement intended to allow us- ers to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited. Link: https://do1.org/10.3897/zse.100.119153.suppl1 Supplementary material 2 Characters Urophonius phylogeny Authors: Andrés A. Ojanguren-Affilastro, Fermin M. Alfaro, Martin J. Ramirez, Bernardino Camoussel- gt-Montolivo, Jaime Pizarro-Araya Data type: doc Copyright notice: This dataset is made available under the Open Database License (http://opendatacommons. org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow us- ers to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited. Link: https://doi.org/10.3897/zse.100.119153.suppl2 zse.pensoft.net Supplementary material 3 Matrix Urophonius phylogeny Authors: Andrés A. Ojanguren-Affilastro, Fermin M. Alfaro, Martin J. Ramirez, Bernardino Camoussei- gt-Montolivo, Jaime Pizarro-Araya Data type: txt Copyright notice: This dataset 1s made available under the Open Database License (http://opendatacommons. org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow us- ers to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited. Link: https://doi.org/10.3897/zse.100.119153.suppl3