Background Culicoides biting midges (Diptera, Ceratopogonidae) are known vectors of avian Haemoproteus parasites. These parasites cause diseases, pathology and even mortality in birds. The diversity of biting midges in Europe is great, but only four Culicoides species are known to be vectors of avian Haemoproteus parasites. In general, our knowledge about the role of the particular Culicoides species in the transmission of Haemoproteus parasites remains insufficient. Information gaps hinder a better understanding of parasite biology and the epizootiology of parasite-caused diseases. The aim of this study was to determine new Culicoides species involved in the transmission of Haemoproteus parasites. Methods Biting midges were collected using a UV trap as well as sticky traps installed in bird nest boxes. Individual parous females were diagnosed for the presence of haemoproteids using both PCR-based and microscopic methods. Results We collected and dissected 232 parous Culicoides females from 9 species using a UV trap and 293 females from 11 species from bird nest boxes. Culicoides obsoletus was the dominant species collected using a UV trap, and Culicoides kibunensis dominated among midges collected in nest boxes. PCR-based screening showed that 5.2% of parous biting midges collected using a UV trap and 4.4% of midges collected from nest boxes were infected with avian haemosporidian parasites. Haemoproteid DNA was detected in C. kibunensis, Culicoides pictipennis, Culicoides punctatus, Culicoides segnis and Culicoides impunctatus females. The sporozoites of Haemoproteus minutus (genetic lineages hTURDUS2 and hTUPHI01) were detected in the salivary glands of two C. kibunensis females using microscopy, and this finding was confirmed by PCR. Conclusions Culicoides kibunensis was detected as a new natural vector of Haemoproteus minutus (hTURDUS2 and hTUPHI01). Haemoproteid DNA was detected in females from five Culicoides species. This study contributes to the epizootiology of avian Haemoproteus infections by specifying Culicoides species as vectors and species that are likely to be responsible for the transmission of haemoproteids in Europe.

BackgroundCulicoides biting midges (Diptera, Ceratopogonidae) are known to be vectors of avian Haemoproteus parasites. These parasites cause disease and pathology in birds. The diversity of biting midges in Europe is great, but only four Culicoides species are known to be vectors of avian Haemoproteus parasites. In general, our knowledge about the role of the particular Culicoides species in transmission of Haemoproteus parasites remains insufficient. Information gaps hinder a better understanding of parasite biology and the epizootiology of parasite-caused diseases. The aim of this study was to determine new ornithophilic Culicoides species potentially involved in local transmission of Haemoproteus parasites. To do this we collected biting midges in bird nest boxes, identified their species and prevalence of haemoproteids in insects as well as in juvenile birds during breeding season.Methods Biting midges were collected from bird nest boxes, identified and parous females were tested individually for the presence of haemoproteids. The blood of juvenile birds was sampled to determine a local transmission of Haemoproteus spp. in the study area. We have used both microscopy and PCR-based methods.ResultsIn all, 293 Culicoides females belonging to 11 species were collected from nest boxes. Culicoides pictipennis, Culicoides segnis and Culicoides kibunensis were determined as dominant species collected using this method. Culicoides kibunensis was found to be infected with Haemoproteus lanii (genetic lineage hRB1), C.pictipennis and Culicoides punctatus− with Haemoproteus minutus (hTUPHI01 and hTURDUS2, respectively), C.segnis− with Haemoproteus majoris (hCWT4), H.minutus (hTURDUS2) and Haemoproteus tartakovskyi (hSISKIN1). From 187 studied juvenile birds 31 were infected with H. majoris (hCWT4, hPARUS1, hPHSIB1, hWW2) and Haemoproteus sp. (hPARUS10) parasites, which are widespread in Europe.Conclusions Our results provided information about the distribution of biting midge species and determined new ornithophilic Culicoides species at the study site. This study contributes to epizootiology of avian Haemoproteus infections by specifying Culicoides species that likely are responsible for the transmission of haemoproteids in Europe. Used method is suitable for better understanding vector ecology and evaluating the role of different blood sucking insects in transmission of haemoproteids in different wild ecosystems.