Unravelling the phylogenetic complexities of Ischyropsalis (Opiliones: Dyspnoi: Ischyropsalididae) in the North Iberian Peninsula, with the description of three new species
Abstract
Ischyropsalis C.L. Koch, 1839 is a genus of harvestmen inhabiting both terrestrial surface habitats and caves in central and southern Europe. The accepted number of species is controversial due to feature similarities and their strong sexual dimorphism. The northern part of the Iberian Peninsula harbours 16 accepted species, seven of which are epigean, while the other nine exclusively live in caves. Some of these species have not been included in the only phylogenetic study conducted on this genus, so their taxonomic status has not yet been evaluated within a phylogenetic framework. In this context, we aim to unravel the phylogenetic relationships among the taxa within this genus, from the northern part of the Iberian Peninsula, by conducting morphological and phylogenetic analyses based on mitochondrial (COI) and nuclear (EF1-α) molecular markers. Our results revealed three previously undescribed species from karstic caves in Cantabria and Asturias, herein described as I. damiani sp. nov., I. impressa Luque & Labrada sp. nov. and I. aguerana Luque & Labrada sp. nov. and a new designation, nodifera-group, for those Iberian species related with I. nodifera.
References
Adrados-González L. 2011. Costa oriental de Asturias. Un Paisaje singular. Adrados. Oviedo.
Akaike H. 1974. A new look at the statistical model identification. IEEE Transactions on Automatic Control 19 (6): 716–723. Available from https://ieeexplore.ieee.org/document/1100705 [accessed 26 Aug. 2026].
Altekar G., Dwarkadas S., Huelsenbeck J.P. & Ronquist F. 2004. Parallel metropolis-coupled Markov chain Monte Carlo for Bayesian phylogenetic inference. Bioinformatics 20: 407–415. https://doi.org/10.1093/bioinformatics/btg427
Angus R.B., Edwards D.B., Luque C.G. & Labrada L. 2012. A chromosomal investigation of some European Leiodidae (Coleoptera), with particular focus on Spanish subterranean Leptodirini. Comparative Cytogenetics 6 (2): 127–139. https://doi.org/10.3897/CompCytogen.v6i2.2575
Ballesteros D., Jiménez-Sánchez M. & García-Sansegundo J. 2013. Patrimonio geológico subterráneo en espacios naturales protegidos: caracterización geomorfológica preliminar de sistemas kársticos profundos en el Parque Nacional de los Picos de Europa (España). In: Vega J., Salazar A., Díaz-Martínez E. & Marchán C. (eds) Patrimonio geológico, un Recurso para el Desarrollo: 361–370. Instituto Geológico y Minero de España. Madrid.
Baró-Pazos J. 2010. Los Hitos de un histórico Conflicto territorial entre Cantabria y el País Vasco: el Caso Agüera (Guriezo) y Trucíos-Turtzioz. Desde sus Orígenes (Siglo XVI) hasta la Sentencia del Tribunal Supremo de 9 de abril de 2008. Regional Government of Cantabria (Consejería de Presidencia y Justicia), Santander.
Chernomor O., Von Haeseler A. & Quang Minh B. 2016. Terrace aware data structure for phylogenomic inference from supermatrices downloaded from supermatrices. Systematic Biology 65 (6): 997–1008. https://doi.org/10.1093/sysbio/syw037
Culver D.C. & Pipan T. 2009. The Biology of Caves and Other Subterranean Habitats. Oxford University Press. https://doi.org/10.1093/oso/9780198820765.001.0001
Culver D.C., Deharveng L., Bedos A., Lewis J.J., Madden M., Reddell J.R., Sket B., Trontelj P. & White D. 2006. The mid-latitude biodiversity ridge in terrestrial cave fauna. Ecography 29: 120–128. https://doi.org/10.1111/j.2005.0906-7590.04435.x
Darriba D., Taboada G.L., Doallo R. & Posada D. 2012. JModelTest 2: more models, new heuristics and parallel computing. Nature Methods 9 (8): 772. https://doi.org/10.1038/nmeth.2109
Dresco E. 1968. Recherches sur les opilions du genre Ischyropsalis (Fam. Ischyropsalidae). VII. Ischyropsalis gigantea sp. nov. Bulletin du Museum national d’Historie naturelle, 2e série 40 (2): 308–314.
Dresco E. 1970. Recherches sur la variabilité et la phylogénie chez les opilions du genre Ischyropsalis C.L. Koch (Fam. Ischyropsalidae), avec création de la famille nouvelle des Sabaconidae. Bulletin du Muséum national d’Histoire naturelle 41: 1200–1213.
Edgar R.C. 2004. MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Research 32 (5): 1792–1797. https://doi.org/10.1093/nar/gkh340
Faille A., Balart-García P., Fresneda J., Bourdeau C. & Ribera I. 2021. A remarkable new genus of Iberian troglobitic Trechodina (Coleoptera: Carabidae: Trechinae: Trechini), with a revisited molecular phylogeny of the subtribe. Annales de la Société entomologique de France (N.S.) 57: 85–106. https://doi.org/10.1080/00379271.2021.1880339
Federación Cántabra de Espeología (FCE). Available from https://www.espeleocantabria.net [accessed 26 Aug. 2026].
Federación Española de Espeología (FEE). Available from https://feespeleo.com [accessed 26 Aug. 2026].
Federación d’Espeoloxía del Principáu d’Asturies (FESPA). Available from https://fespasturies.com [accessed 26 Aug. 2026].
Folmer O., Black M., Hoeh W., Lutz R. & Vrijenhoek R. 1994. DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular Marine Biology and Biotechnology 3 (5): 294–299.
Fresneda F., Giachino P.M., Salgado J.M., Faille A., Bourdeau C., Cieslak A. & Ribera I. 2024. A phylogenetic classification of Leptodirini (Coleoptera, Leiodidae, Cholevinae). Memorie della Società Entomologica Italiana 101: 3–936. https://doi.org/10.4081/memoriesei.2024.3
García de la Noceda C., De Toledo F.O. & Del Pozo Gómez M. 1999. Programa de Actualización del Inventario Hidrogeológico (PAIH). Análisis del Conocimiento actual Evaluación y Programación de Estudios en las Cuencas intercomunitarias. Instituto Geológico y Minero de España, Madrid.
Garzia M., Berrilli E., Lunghi E., Coppari L., Delcour N. & Salvi D. 2025. Genetic diversity and distribution of Italian cave crickets (Dolichopoda): toward a better understanding of lineage structure. Animals 15 (16): 2429. https://doi.org/10.3390/ani15162429
Hastings W.K. 1970. Monte Carlo sampling methods using Markov chains and their applications. Biometrika 57 (1): 97–103. https://doi.org/10.2307/2334940
Hedin M. 1997. Molecular phylogenetics at the population/species interface in cave spiders of the southern Appalachians (Araneae: Nesticidae: Nesticus). Molecular Biology and Evolution 14 (3): 309–324. https://doi.org/10.1093/oxfordjournals.molbev.a025766
Hedin M., Derkarabetian S., McCormack M., Richart C. & Shultz J.W. 2010. The phylogenetic utility of the nuclear protein-coding gene EF1-α for resolving recent divergences in Opiliones, emphasizing intron evolution. The Journal of Arachnology 38: 9–20. https://doi.org/10.1636/ha09-49.1
Helversen O. Von. & Martens J. 1972. Unrichtige Fundort-Angaben in der Arachniden-Sammlung Roewer. Senckenbergiana Biologica 53: 109–123.
Hoang D.T., Chernomor O., Von Haeseler A., Quang Minh B. & Sy Vinh L. 2017. UFBoot2: omproving the ultrafast bootstrap approximation. Molecular Biology and Evolution 30 (5): 1188–1195. https://doi.org/10.1093/molbev/msx281
Kearse M., Moir R., Wilson A., Stones-Havas S., Cheung M., Sturrock S., Buxton S., Cooper A., Markowitz S., Duran C., Thierer T., Ashton B., Meintjes P. & Drummond A. 2012. Geneious Basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics 28 (12): 1647–1649. https://doi.org/10.1093/bioinformatics/bts199
Kury A.B., Mendes A.C., Cardoso L., Kury M.S., Granado A. de A., Giribet G., Cruz-López J.A., Longhorn S.J., Medrano M., Oliveira A.B.R. de, Kury I.S. & Souza-Kury M.A. 2024. World Catalogue of Opiliones. WCO-Lite version 2.7. Available from https://wcolite.com/ [accessed 26 Aug. 2026].
Labrada L., Salgado J.M. & Luque C.G. 2010. Fauna invertebrada de las cavidades de Cantabria. Locustella, Anuario de la Naturaleza de Cantabria 7: 29–43. Available from http://www.impress.es/docs/publicaciones/Locustella_2010.pdf [accessed 26 Aug. 2026].
Labrada L., Tosal L., Gutiérrez M., Ranero S.L., Tosal E. & Luque C.G. in press. Life in the dark: Preliminary list of the strictly adapted cave fauna in Cantabria region (Northern Spain). Bulletin of the Cantabrian Cave Research & Caving Federation 18, ver. 2026.1.1 (last updated on 10 Oct. 2026). Santander.
Leray M., Yang J.Y., Meyer C.P., Mills S.C., Agudelo N., Ranwez V., Boehm J.T. & Machida R.J. 2013. A new versatile primer set targeting a short fragment of the mitochondrial COI region for metabarcoding metazoan diversity: application for characterizing coral reef fish gut contents. Zoology 10 (34): 1–14. https://doi.org/10.1186/1742-9994-10-34
Luque C.G. 1991. Los Ischyropsalidoidea de la Cornisa Cantábrica: Cantabria (Arachnida: Opiliones: Palpatores: Ischyropsalididae y Sabaconidae). Arquenas 1: 1–96.
Luque C.G. 1992. Contribución al conocimiento de los Opiliones epigeos e hipogeos de Cantabria (España). Primeras aportaciones al catálogo (Arachnida: Opiliones: Palpatores). Proceedings of the 5th Spanish Meeting of Speleology, Camargo-Santander, Spain 1–4 November 1990: 143–157 + errata sheet. Cantabrian Caving Federation, Santander.
Luque C.G. & Labrada L. 2012. A new cave-dwelling endemic Ischyropsalis C.L. Koch, 1839 (Opiliones: Dyspnoi: Ischyropsalididae) from the karstic region of Cantabria (Spain). Zootaxa 3506: 26–42. https://doi.org/10.11646/zootaxa.3506.1.2
Luque C.G. & Labrada L. 2015. Organizing biogeomorphic information aimed at the planning and management of protected areas in Cantabria (Spain). The case of El Pendo and Peñajorao Caves – an example of Special Interest Area and UNESCO World Heritage Site. Boletín de la real Sociedad española de Historia natural (Sección Geológica) 109: 5–25. Available from http://www.rsehn.es/publicaciones-geologica/art278 [accessed 26 Aug. 2026].
Luque C.G. & Labrada L. 2017. Nuevas localizaciones de la especie Cantabrodesmus lorioli Mauriès, 1971 en Cantabria, España (Polydesmida, Diplopoda, Chelodesmidae). Boletín de la real Sociedad española de Historia natural (Sección Biológica) 111: 51–60. Available from http://www.rsehn.es/publicaciones-biologica/art370 [accessed 26 Aug. 2026].
Luque C.G. & Labrada L. 2022. Paisaje, territorio y patrimonio natural. In: Fernández Vega P.A. (coord.) Historia y Patrimonio de Cantabria: 1–40. Regional Government of Cantabria (Consejería de Educación). Santander.
Mammola S., Schönhofer A.L. & Isaia M. 2019. Tracking the ice: subterranean harvestmen distribution matches ancient glacier margins. Journal of Zoological Systematics and Evolutionary Research 57: 548–554. https://doi.org/10.1111/jzs.12264
Martens J. 1969a. Die Sekretdarbietung während des Paarungsverhaltens von Ischyropsalis C.L. Koch (Opiliones). Zeitschrift fur Tierpsycholologie 26: 513–523.
Martens J. 1969b. Die Abgrenzung von Biospezies auf biologisch-ethologischer und morphologischer Grundlage am Beispiel der Gattung Ischyropsalis C.L. Koch 1839 (Opiliones, Ischyropsalididae). Zoologische Jahrbücher Abteilung für Systematik 96: 133–264. Available from https://www.researchgate.net/publication/292124925 [accessed 26 Aug. 2026].
Martens J. 1978. Spinnentiere, Arachnida – Weberknechte, Opiliones. Die Tierwelt Deutschlands 64: 1–464.
Metropolis N., Rosenbluth A.W., Rosenbluth M.N., Telle A.H. & Teller E. 1953. Equation of state calculations by fast computing machines. Journal of Chemical Physics 21: 1087–1092. https://doi.org/10.1063/1.1699114
Minh B.Q., Nguyen M.A.T. & Von Haeseler A. 2013. Ultrafast approximation for phylogenetic bootstrap. Molecular Biology and Evolution 30 (5): 1188–1195. https://doi.org/10.1093/molbev/mst024
Moore G.W., Goodman M. & Barnabas J. 1973. An iterative approach from the standpoint of the additive hypothesis to the dendrogram problem posed by molecular data sets. Journal of Theoretical Biology 38 (3): 423–457. https://doi.org/10.1016/0022- 5193(73)90251-8
Nguyen L.T., Schmidt H.A., Von Haeseler A. & Minh B.Q. 2015. IQ-TREE: A fast and effective stochastic algorithm for estimating maximum-likelihood phylogenies. Molecular Biology and Evolution 32 (1): 268–274. https://doi.org/10.1093/molbev/msu300
Pinto-da-Rocha R., Machado G. & Giribet G. (eds). 2007. Harvestmen: The Biology of Opiliones. Harvard University Press, Cambridge, UK.
Prieto C.E. 1990. The genus Ischyropsalis C.L. Koch (Opiliones, Ischyropsalididae) on the Iberian Peninsula. I. Non-troglobitic species. Acta Zoologica Fennica 190: 315–320.
Prieto C., Malumbres-Olarte J., Luque C.G., Labrada L. & Schönhofer A. 2013. The Ischyropsalis radiation from the Basque-Cantabrian region: Multiple colonization of caves or recolonization of epigean habitats? III Encuentro Ibérico de Biología Subterránea, Barcelona (Poster). Available from https://www.researchgate.net/publication/321252883 [accessed 26 Aug. 2026].
Rambaut A., Drummond A.J., Xie D., Baelz G. & Suchard M.A. 2018. Posterior summarization in Bayesian phylogenetics using Tracer 1.7. Systematic Biology 67 (5): 901–904. https://doi.org/10.1093/sysbio/syy032
Ribera I., Fresneda J., Bucur R., Izquierdo A., Vogler A.P., Salgado J.M. & Cieslak A. 2010. Ancient origin of a Western Mediterranean radiation of subterranean beetles. BMC Evolutionary Biology 10 (29): 1–14. https://doi.org/10.1186/1471-2148-10-29
Robinson D.F. 1971. Comparison of labeled trees with valency three. Journal of Combinatorial Theory, Series B 11 (2): 105–119. https://doi.org/10.1016/0095-8956(71)90020-7
Roewer C.F. 1950. Über Ischyropsalididae and Trogulidae: Weitere Weberknechte XV. Senckenbergiana 28: 7–57.
Ronquist F., Teslenko M., Van Der Mark P., Ayres D.L., Darling A., Höhna S., Larget B., Liu L., Suchard M.A. & Huelsenbeck J.P. 2012. MrBayes 3.2: Efficient bayesian phylogenetic inference and model choice across a large model space. Systematic Biology 61 (3): 539–542. https://doi.org/10.1093/sysbio/sys029
Rozwałka R., Mazur A. & Staręga W.K. 2012. Distribution of harvestmen of the genus Ischyropsalis C.L. Koch (Arachnida: Opiliones) in Poland. Fragmenta Faunistica 55 (1): 11–18. https://doi.org/10.3161/00159301FF2012.55.1.011
Ruiz-Cobo J., Muñoz-Fernández E. & Luque C.G. 2008. Revisión del complejo arqueológico del karst de la Baja (Laredo, Cantabria). Sautuola, Journal of Institute of Prehistory and Archaeology of Santander 14: 41–58. Available from https://institutosautuola.es/descargas-sautuola/ [accessed 26 Aug. 2026].
Salgado J.M., Luque C.G., Labrada L., Fresneda J. & Ribera I. 2012. Revisión del género Cantabrogeus Salgado, 2000, con la descripción de tres nuevas especies hipogeas endémicas de la Cordillera Cantábrica (Coleoptera, Leiodidae, Cholevinae, Leptodirini). Animal Biodiversity and Conservation 35 (1): 27–50.https://doi.org/10.32800/abc.2012.35.0027
Santos J., Redondo J.M., González-Gutiérrez R.B. & Gómez-Villar A. 2013. Applying the AABR method to reconstruct equilibrium-line altitudes from the last glacial maximum in the Cantabrian Mountains (SW Europe). Palaeogeography, Palaeoclimatology & Palaeoecology 387: 185–199. https://doi.org/10.1016/j.palaeo.2013.07.025
Schmidt M., Achterhold K., Pfeiffer F. & Melzer R.R. 2024. Kinematics of elongate harvestmen chelicerae: comparative range of motion analysis in extant Ischyropsalis (Dyspnoi, Opiliones). Zoology 167: 1–11. https://doi.org/10.1016/j.zool.2024.126219
Schönhofer A.L. 2013. A taxonomic catalogue of the Dyspnoi Hansen & Sørensen, 1904 (Arachnida: Opiliones). Zootaxa 3679: 1–68. https://doi.org/10.11646/zootaxa.3679.1.1
Schönhofer A.L. & Martens J. 2010. On the identity of Ischyropsalis dentipalpis Canestrini, 1872 and description of Ischyropsalis lithoclasica sp. n. (Opiliones: Ischyropsalididae). Zootaxa 2613 (1): 1–14. https://doi.org/10.11646/zootaxa.2613.1.1
Schönhofer A.L., Vernesi C., Martens J. & Hedin M. 2015. Molecular phylogeny, biogeographic history, and evolution of cave-dwelling taxa in the European harvestman genus Ischyropsalis (Opiliones: Dyspnoi). The Journal of Arachnology 43: 40–53. https://doi.org/10.1636/H14-39.1
Sendra A. (ed.) 2023. Habitantes de la Oscuridad. Fauna Ibero-balear de las Cuevas. Sociedad Entomológica Aragonesa, Zaragoza.
Serrano E., González-Trueba J.J., Pellitero R. & Gómez-Lende M. 2013. Quaternary glacial evolution in the Central Cantabrian Mountains (Northern Spain). Geomorphology 196: 65–82. https://doi.org/10.1016/j.geomorph.2012.05.001
Serrano E., González-Trueba J.J., Pellitero R. & Gómez-Lende M. 2017. Quaternary glacial history of the Cantabrian Mountains of northern Spain: a new synthesis. Geological Society London 433: 55–85. https://doi.org/10.1144/SP433.8
Tavaré S. 1986. Some probabilistic and statistical problems in the analysis of DNA sequences. In: Miura E.R.M. (ed.) Lectures on Mathematics in the Lfe Sciences. American Mathematical Society 17 (2): 57–86..
Thorell T.1876. Sopra alcuni Opilioni (Phalangidea) d’Europa e dell’Asia occidentale, con un quadro dei generi europei di quest’Ordine. Annali del Museo Civico di Storia Naturale di Genova 1 (8): 452–508. https://doi.org/10.5962/bhl.part.10418
Trifinopoulos J., Nguyen L.T., von Haeseler A. & Minh B.Q. 2016. W-IQ-TREE: a fast online phylogenetic tool for maximum likelihood analysis. Nucleic Acids Research 44 (1): 232–235. https://doi.org/10.1093/NAR/GKW256
Vink C.J., Thomas S.M., Paquin P., Hayashi C.Y. & Hedin M. 2005. The effects of preservatives and temperatures on arachnids DNA. Invertebrate Systematics 19: 99–104. https://doi.org/10.1071/IS04039
Waterhouse A.M., Procter J.B., Martin D.M.A., Clamp M. & Barton G.J. 2009. Jalview Version 2-A multiple sequence alignment editor and analysis workbench. Bioinformatics 25 (9): 1189–1191. https://doi.org/10.1093/bioinformatics/btp03
Yang Z. 1994. Maximum likelihood phylogenetic estimation from DNA sequences with variable rates oversites: approximate methods. Journal of Molecular Evolution 39: 306–314. https://doi.org/10.1007/BF00160154
Copyright (c) 2026 Ricardo López-Alonso, Esteban Pascual-Parra, Lucía Labrada, Carlos G. Luque, Eduardo Cires, Andrés Arias, Claudia González-Toral

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