Water diviners described: six new species of the Niphargus aquilex (Crustacea, Amphipoda) complex

Keywords: cryptic diversity, Niphargus, integrative taxonomy, groundwater fauna, species complex

Abstract

With the onset of molecular taxonomy, it has been recognized that groundwater crustaceans in the family Niphargidae Bousfield, 1977 (Crustacea, Amphipoda) can demonstrate either highly variable or static phenotypes. Thus, it is recommended that new species descriptions follow an integrative taxonomic approach, considering morphotaxonomic and molecular characteristics to establish new taxa. The morphospecies Niphargus aquilex Schiødte, 1855 has its main distribution area in Central Europe, spanning southern England, the northern half of France, the Benelux countries, Germany and the Czech Republic. Genetic analyses have shown that it comprises a highly diverse species complex, including amongst others the well-recognized morphospecies Niphargus schellenbergi S. Karaman, 1932. In this study, we describe six new species based on available adult males and females – Niphargus luxemburgensis Weber, Weigand & Brad sp. nov., N. palatinensis Weber & Brad sp. nov., N. normandiensis Weber & Brad sp. nov., N. wasgauensis Weber & Brad sp. nov., N. saraviensis Weber & Brad sp. nov., N. lotharingiensis Weber & Brad sp. nov. – taking a re-evaluated published genetic backbone as a further reference into account. The six newly described species appeared pseudocryptic, as minor morphological differences were noticed. A short discussion of the morphological and molecular delimitations is provided.

References

Aparicio-Puerta E., Gómez-Martín C., Giannoukakos S., Medina J.M., Scheepbouwer C., García-Moreno A., ... & Hackenberg M. 2022. sRNAbench and sRNAtoolbox 2022 update: accurate miRNA and sncRNA profiling for model and non-model organisms. Nucleic Acids Research 50 (W1): W710–W717. https://doi.org/10.1093/nar/gkac363

Astrin J.J. & Stüben P.E. 2008. Phylogeny in cryptic weevils: molecules, morphology and new genera of western Palaearctic Cryptorhynchinae (Coleoptera: Curculionidae). Invertebrate Systematics 22 (5): 503–522. https://doi.org/10.1071/IS07057

Brad T., Fišer C., Flot J.-F. & Sarbu S.M. 2015. Niphargus dancaui sp. nov. (Amphipoda, Niphargidae) – a new species thriving in sulfidic groundwaters in southeastern Romania. European Journal of Taxonomy 164: 1–28. https://doi.org/10.5852/ejt.2015.164

Delić T., Trontelj P., Rendoš M. & Fišer C. 2017. The importance of naming cryptic species and the conservation of endemic subterranean amphipods. Scientific Reports 7: 3391. https://doi.org/10.1038/s41598-017-02938-z

Dobat K. 1968. Mitteilung über die aquatile Fauna einiger Höhlen der Schwäbischen Alb. Mitteilungen des Verbandes der deutschen Höhlen- und Karstforscher e. V. 14 (11): 31–33.

Dobreanu E., Manolache C. & Puscariu V. 1953. Noi specii de Amphipode freatice din R.P.R. Bulletin Stiintific, Sectiunea de Stiinte Biologice, Agronomice, Geologice si Geografice 5 (3): 603–616.

Eme D., Zagmajster M., Delić T., Fišer C., Flot J.-F., Konecny-Dupré L., Snaebjörn P., Stoch F., Zakšek V., Douady C.J. & Malard F. 2018. Do cryptic species matter in macroecology? Sequencing European groundwater crustaceans yields smaller ranges but does not challenge biodiversity determinants. Ecography 41 (2): 424–436. https://doi.org/10.1111/ecog.02683

Esmaeili-Rineh S., Sari A., Fišer C. & Bargrizaneh Z. 2017. Completion of molecular taxonomy: description of four amphipod species (Crustacea: Amphipoda: Niphargidae) from Iran and release of database for morphological taxonomy. Zoologischer Anzeiger 271: 57–79. https://doi.org/10.1016/j.jcz.2017.04.009

Fišer C. & Zagmajster M. 2009. Cryptic species from cryptic space: the case of Niphargus fongi sp. n. (Amphipoda, Niphargidae). Crustaceana 82 (5): 593–614. https://doi.org/10.1163/156854009X407704

Fišer C., Sket B., Turjak M. & Trontelj P. 2009a. Public online databases as a tool of collaborative taxonomy: a case study on subterranean amphipods. Zootaxa 2095 (1): 47–56. https://doi.org/10.11646/zootaxa.2095.1.5

Fišer C., Trontelj P., Luštrik R. & Sket B. 2009b. Towards a unified taxonomy of Niphargus (Crustacea: Amphipoda): a review of morphological variability. Zootaxa 2061 (1): 1–22. https://doi.org/10.11646/zootaxa.2061.1.1

Fišer C., Konec M., Alther R., Švara V. & Altermatt F. 2017. Taxonomic, phylogenetic and ecological diversity of Niphargus (Amphipoda: Crustacea) in the Hölloch cave system (Switzerland). Systematics and Biodiversity 15 (3) 218–237. https://doi.org/10.1080/14772000.2016.1249112

Fišer C., Alther R., Zakšek V., Borko Š., Fuchs A. & Altermatt F. 2018. Translating Niphargus barcodes from Switzerland into taxonomy with a description of two new species (Amphipoda, Niphargidae). ZooKeys 760: 113–141. https://doi.org/10.3897/zookeys.760.24978

Flot J.-F., Wörheide G. & Dattagupta S. 2010. Unsuspected diversity of Niphargus amphipods in the chemoautotrophic cave ecosystem of Frasassi, central Italy. BMC Evolutionary Biology 10: 171. https://doi.org/10/dbpkg8

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.

Gerecke R., Stoch F., Meisch C. & Schrankel I. 2005. Die Fauna der Quellen und des hyporheischen Interstitials in Luxembourg. Ferrantia 41. Museum of Natural History of Luxembourg, Luxembourg.

Horton T., De Broyer C., Bellan-Santini D., Coleman C.O., Copilaș-Ciocianu D., Corbari L., Daneliya M.E., Dauvin J.-C., Decock W., Fanini L., Fišer C., Gasca R., Grabowski M., Guerra-García J.M., Hendrycks E.A., Hughes L.E., Jaume D., Kim Y.-H., King R.A., Lo Brutto S., Lörz A.-N., Mamos T., Serejo C.S., Senna A.R., Souza-Filho J.F., Tandberg A.H.S., Thurston M.H., Vader W., Väinölä R., Valls Domedel G., Vandepitte L., Vanhoorne B., Vonk R., White K.N. & Zeidler W. 2023. The World Amphipoda Database: history and progress. Records of the Australian Museum 75 (4): 329–342. https://doi.org/10.3853/j.2201-4349.75.2023.1875

Husmann S. 1976. Studies on subterranean drift of stygobiont crustaceans (Niphargus, Crangonyx, Graeteriella). International Journal of Speleology 8: 81–92. https://doi.org/10.5038/1827-806X.8.1.7

Inkscape Project. 2017. Inkscape (Version 0.92) [Computer software]. Available from https://inkscape.org [accessed 14 Aug. 2025].

Jörger K.M. & Schrödl M. 2014. How to use CAOS software for taxonomy? A quick guide to extract diagnostic nucleotides or amino acids for species descriptions. Spixiana 37 (1): 21–26.

Karaman G.S. 1980. Contribution to the knowledge of the Amphipoda 113. Redescription of Niphargus aquilex Schiödte and its distribution in Great Britain. Biosistematika 6 (2): 175–185.

Karaman G.S. 1993. Fauna d’Italia (31): Crustacea Amphipoda (d’aqua dolce). Edizioni Calderini, Bologna.

Karaman G.S. 2017. New data on two subterranean species of the family Niphargidae from Spain, Niphargus gallicus Schell., 1935 and N. delamarei Ruffo, 1954 (contribution to the knowledge of the Amphipoda 282). Contributions, Section of Natural, Mathematical and Biotechnical Sciences 36 (2): 105–120.

Karaman S. 1932. 5. Beitrag zur Kenntnis der Süsswasser-Amphipoden. Prirodoslovne razprave 2: 179–232.

Knowlton N. 1993. Sibling species in the sea. Annual Review of Ecology and Systematics 24 (1): 189–216. https://doi.org/10.1146/annurev.es.24.110193.001201

Kumar S., Stecher G., Li M., Knyaz C. & Tamura K. 2018. MEGA X: molecular evolutionary genetics analysis across computing platforms. Molecular Biology and Evolution 35 (6): 1547–1549. https://doi.org/10.1093/molbev/msy096

Kureck A. 1967. Über die tagesperiodische Ausdrift von Niphargus aquilex schellenbergi KARAMAN aus Quellen. Zeitschrift für Morphologie und Ökologie der Tiere 58: 247–262. https://doi.org/10.1007/BF00407379

Luttikhuizen P.C. & Dekker R. 2010. Pseudo-cryptic species Arenicola defodiens and Arenicola marina (Polychaeta: Arenicolidae) in Wadden Sea, North Sea and Skagerrak: Morphological and molecular variation. Journal of Sea Research 63 (1): 17–23. https://doi.org/10.1016/j.seares.2009.09.001

McInerney C.E., Maurice L., Robertson A.L., Knight L.R.F.D., Arnscheidt J., Venditti C., Dooley J.S.G., Mathers T., Matthijs S., Erikkson K., Proudlove G.S. & Hänfling B. 2014. The ancient Britons: Groundwater fauna survived extreme climate changes over tens of millions of years across NW Europe. Molecular Ecology 23: 1153–1166. https://doi.org/10.1111/mec.12664

Puillandre N., Brouillet S. & Achaz G. 2020. ASAP: assemble species by automatic partitioning. Molecular Ecology Resources 21: 609–620. https://doi.org/10.1111/1755-0998.13281

QGIS Development Team. 2025. QGIS Geographic Information System (Version 3.x) [Computer software]. QGIS Association. Available from https://qgis.org [accessed 14 Aug. 2025].

Sayers E.W., Cavanaugh M., Clark K., Pruitt K.D., Schoch C.L., Sherry S.T. & Karsch-Mizrachi I. 2021. GenBank. Nucleic Acids Research 49: D92–D96. https://doi.org/10.1093/nar/gkaa1023

Schellenberg A. 1932. Deutsche subterrane Amphipoden. Zoologischer Anzeiger 99 (11/12): 311–323.

Schellenberg A. 1933. Weitere deutsche und ausländische Niphargiden. Zoologischer Anzeiger 102: 22–33.

Schiödte J.C. 1855. Om den i England opdagede art af hulekrebs-slægten Niphargus. Oversigt over det Kongelige danske Videnskabernes Selskabs Forhandlinger og dets Medlemmers Arbeider i Aaret 1855 (7–8): 349–351.

Sket B. 1999. Niphargus aquilex dobati ssp. n. (Crustacea) from the karst of Slovenia. Mitteilungen des Verbandes der deutschen Höhlen- und Karstforscher 45: 54–56.

Straškraba M. 1972. Les groupement des espèces du genre Niphargus (sensu lato). In: Ruffo S. (ed.) Actes du premier colloque international sur le genre Niphargus: 85–90. Museo Civico di Storia Naturale di Verona.

Thienemann A. 1922. Hydrobiologische Untersuchungen an Quellen. Archiv für Hydrobiologie 14: 151–190.

Verovnik R., Sket B. & Trontelj P. 2005. The colonization of Europe by the freshwater crustacean Asellus aquaticus (Crustacea: Isopoda) proceeded from ancient refugia and was directed by habitat connectivity. Molecular Ecology 14 (14): 4355–4369. https://doi.org/10.1111/j.1365-294X.2005.02745.x

Weber D. 1988. Die Höhlenfauna und -flora des Höhlenkatastergebietes Rheinland-Pfalz/Saarland. Abhandlungen zur Karst- und Höhlenkunde 22. Verband der deutschen Höhlen- und Karstforscher, München.

Weber D. 1989. Die Höhlenfauna und -flora des Höhlenkatastergebietes Rheinland-Pfalz/Saarland, 2. Teil. Abhandlungen zur Karst- und Höhlenkunde 23. Verband der deutschen Höhlen- und Karstforscher, München.

Weber D. & Weigand A.M. 2023. Groundwater amphipods of the hyporheic interstitial: a case study from Luxembourg and the greater region. Diversity 15 (3): 411. https://doi.org/10.3390/d15030411

Weber D., Flot J.-F., Frantz A.C. & Weigand A.M. 2022. Molecular analyses of groundwater amphipods (Crustacea: Niphargidae) from Luxembourg: new species reveal limitations of morphology-based checklists. Zootaxa 5222 (6): 501–533. https://doi.org/10.11646/zootaxa.5222.6.1

Weber D., Brad T., Weigand A. & Flot J.-F. 2023. Water diviners multiplied: cryptic diversity in the Niphargus aquilex species complex in Northern Europe. Preprint BioRxiv 2023.08.13.553147. https://doi.org/10.1101/2023.08.13.553147

Werno A. & Weber D. 2008. ‘Zillas Felsenkeller’ in Nunkirchen (Saarland), ein künstlicher Hohlraum mit herausragender Evertebratenfauna. Abhandlungen der Delattinia 34: 139–146.

Westwood J.O. 1853. Notice of the discovery in England of a new genus and species of Amphipodous Crustacea, the Niphargus stygius of Schiödte. Proceedings of the Linnean Society of London 2: 218–219.

Wrześniowski A. 1890. Über drei unterirdische Gammariden. Zeitschrift für wissenschaftliche Zoologie 50: 600–724.

Published
2025-08-25
How to Cite
Weber, D., Brad, T., & Weigand, A. M. (2025). Water diviners described: six new species of the Niphargus aquilex (Crustacea, Amphipoda) complex. European Journal of Taxonomy, 1011(1), 1-79. https://doi.org/10.5852/ejt.2025.1011.3023
Section
Monograph