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Background   Culicoides  biting midges (Diptera: Ceratopogonidae) cause biting nuisance to livestock and humans and are vectors of a range of pathogens of medical and veterinary importance. Despite their economic significance, the delineation and identification of species where only morphology is considered, as well as the evolutionary relationships between species within this genus remains problematic. In recent years molecular barcoding has assisted substantially in the identification of biting midges in the multiple entomological survey projects which were initiated in many European countries following the bluetongue outbreak in 2006–2009. These studies revealed potentially new species and “species-complexes” with large genetic and morphological variability. Here we use molecular barcoding, together with morphological analysis, to study subgenus  Culicoides  Latreille from Scandinavia with focus on three potentially new species.    Methods  Biting midges were collected at various sites in Denmark and Sweden.  Culicoides  specimens were described by variation of a fragment of their cytochrome c oxidase subunit 1 (COI) gene sequence and wing, palp and antennal characters.    Results  It is shown that three new species initially separated by DNA barcoding with mitochondrial COI can be distinguished by morphological characters. In this context a key to Scandinavian subgenus  Culicoides  using wing and maxillary palp characters is presented. The key is including the three new species  Culicoides boyi ,  Culicoides selandicus  and  Culicoides kalix .    Conclusion  Three new species of  Culicoides  biting midges were identified and could be identified by both molecular and morphological differences. Evaluation of differences between and within taxa of biting midges using COI barcode yielded a rough estimate of species delineation; interspecies differences across  Culicoides  subgenera approaches 20%, whereas intraspecies differences are below 4% and in most cases below 1%.    Electronic supplementary material  The online version of this article (doi:10.1186/s13071-015-0750-4) contains supplementary material, which is available to authorized users.

parasites and vectors

Delineation of Culicoides species by morphology and barcode exemplified by three new species of the subgenus Culicoides (Diptera: Ceratopogonidae) from Scandinavia