Demographic history, range size and habitat preferences of the groundwater amphipod Niphargus puteanus (C.L. Koch in Panzer, 1836) Dieter Weber1,2, Jean-François Flot1,3, Hannah Weigand2, Alexander M. Weigand2* 1 Université Libre de Bruxelles, Evolutionary Biology & Ecology group, Avenue F.D. Roosevelt 50, B- 1050 Brussels - Belgium 2 Musée National d'Histoire Naturelle Luxembourg, Section de zoologie, 25 Rue Münster, L-2160 Luxembourg, Luxembourg 3 Interuniversity Institute of Bioinformatics in Brussels – (IB)2, ULB-VUB, La Plaine Campus, Triomflaan, C building, 6th floor, CP 263, 1050 Brussels, Belgium *corresponding author:
[email protected] Abstract Niphargus puteanus is the oldest described species of its genus and, in the past, was used as a taxonomic annotation for any subterranean amphipod record. For that reason, no clear knowledge exists about its actual range size and habitat preferences. We here applied a molecular taxonomic and phylogeographical approach based on mitochondrial and nuclear DNA to shed light on its distribution and to infer its demographic history. Furthermore, we analysed aquifer types and water flow regimes to provide a clearer picture of the species’ ecological requirements. Our results indicate that N. puteanus is widely distributed north of the Alps, having its core range in the geomorphological natural region of the ‘South German Scarplands’ (SGS). Additionally, isolated satellite populations exist in the Taunus and the Sauerland, and two single individuals were collected in Luxembourg and in Austria, respectively. The species’ maximal distribution range reaches 756 km between the two single-specimen records and 371 km within the SGS. A very high haplotype diversity was observed, revealing the presence of seven haplotype groups.