The Carabus subgenus Hygrocarabus contains two taxa: C. variolosus and C. nodulosus, the species or subspecies status of which is handled far from uniform in the literature. Both taxa show a similar morphology, the shape of the tip of the aedeagus provides a reliable morphological marker for identification. We analysed two mitochondrial gene parts (COI-5’ and COI-3’) and a nuclear one (ITS2). High diversity was found showing specific geographical patterns. Introgressive hybridisation was detected but interpreted not as an argument for subspecies status because high genetic distances indicated that it must have taken place in former times. In a laboratory hybridisation experiment, the male did not accept the female of the other taxon, supporting the conclusion that these are separate species. A series of refuges was expected for the period of ice ages. Although only the taxon C. variolosus is listed in Annex II and IV of the EU Habitats Directive, C. nodulosus also falls under this listing, as at the time of including the species into the Annexes in 2004, the two taxa were considered subspecies and hence the listing would include both, independent of later taxonomic revisions.

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Starting point of this study was the problem considering the status of Carabus variolous and C. nodulosus: French and some German authors classified them as species, most German authors as subspecies and Casale as semispecies. We performed analyses of mitochondrial genes (COI-5', COI-3', ND5) as well as nuclear ones (ITS2, wingless) and analysed the DNA sequences using Seqotron, CLUSTALX (editing and alignment), MEGA, DNAML, SplitsTree (phylogeny and network), 4SALE (compensatory base changes), and BEAST (coalescence). We could study specimens from all regions except the western most part of the distribution area (Massif Central and French Jura, France). The mitochondrial DNA data resulted in a geographic pattern of high diversity within both taxa indicating a series of glacial refuges. In addition, a considerably large area was found were introgressive hybridisation took place in the past - at least two times by nodulosus of different regions into variolosus. The nuclear DNA data show a clear and constant difference between both taxa. In consequence, this complex of forms may be characterised as semispecies from an evolutionary viewpoint but taxonomically as one species because of hybridisation and the lack of compensatory bases changes.