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Background   Culicoides  (Diptera: Ceratopogonidae) is a genus of small biting midges (also known as “no-see ums”) that currently includes 1368 described species. They are proven or suspected vectors for important pathogens affecting animals such as bluetongue virus (BTV) and Schmallenberg virus (SBV). Currently little information is available on the species of  Culicoides  present in Serbia. Thus, the aim of this study was to examine species diversity, host preference and the presence of BTV and SBV RNA in  Culicoides  from the Stara Planina Nature Park in south-eastern Serbia.    Results  In total 19,887 individual  Culicoides  were collected during three nights of trapping at two farm sites and pooled into six groups (Obsoletus group, Pulicaris group, “Others” group and further each group according to the blood-feeding status to freshly engorged and non-engorged). Species identification was done on subsamples of 592 individual  Culicoides  specimens by morphological and molecular methods (MALDI-TOF mass spectrometry and PCR/sequencing). At least 22  Culicoides  species were detected. Four animal species (cow, sheep, goat and common blackbird) as well as humans were identified as hosts of  Culicoides  biting midges. The screening of 8291  Culicoides  specimens in 99 pools for the presence of BTV and SBV RNA by reverse-transcription quantitative PCR were negative.    Conclusions  The biodiversity of  Culicoides  species in the natural reserve Stara Planina was high with at least 22 species present. The presence of  C. imicola  Kieffer was not recorded in this area.  Culicoides  showed opportunistic feeding behaviour as determined by host preference. The absence of SBV and BTV viral RNA correlates with the absence of clinical disease in the field during the time of sampling. These data are the direct outcome of a training programme within the Institutional Partnership Project “AMSAR: Arbovirus monitoring, research and surveillance-capacity building on mosquitoes and biting midges” funded by the programme SCOPES of the Swiss National Science Foundation.

Species diversity, host preference and arbovirus detection of Culicoides (Diptera: Ceratopogonidae) in south-eastern Serbia