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Background  Epizootic hemorrhagic disease virus (EHDV) is an  Orbivirus  of veterinary importance which is transmitted by biting midges of the genus  Culicoides  (Diptera: Ceratopogonidae) to ruminants.  Culicoides sonorensis  Wirth & Jones, the only confirmed vector of EHDV in the USA, is rare in the southeastern states where transmission persists, suggesting that other  Culicoides  species transmit EHDV in this region. The present study aimed to determine which  Culicoides  species transmitted EHDV in Florida and Alabama, two states in the southeastern USA. Viral RNA was detected in field-collected midges using molecular methods. These data are presented alongside data on  Culicoides  blood meal analysis, white-tailed deer ( Odocoileus virginianus ) aspiration, and seasonality to demonstrate an interaction between potential vector species and EHDV hosts.    Results  Out of 661 pools tested, 20 pools were positive for EHDV viral RNA, including six pools from  Culicoides stellifer  (Coquillett) and 14 pools from  Culicoides venustus  Hoffman. The overall infection rate was 0.06% for  C. stellifer  and 2.18% for  C. venustus . No positive pools were identified for a further 17 species. Serotypes identified in  Culicoides  included EHDV-2, EHDV-6, and coinfections of EHDV-2 and EHDV-6 and were identified in similar proportions to serotypes in deer at 3 of 4 deer farms. Viral detections conducted in Alabama also identified one positive pool of  C. venustus . Blood meal analysis revealed that both  Culicoides  species fed on white-tailed deer (verified through aspiration), fallow deer, and elk, species for which EHDV viremia has been documented. Seasonality data indicated that both species were present throughout the period in which viral transmission occurred to EHDV hosts in 2016 in addition to the 2017 epizootic.    Conclusions  Our finding of EHDV positive pools of field-collected  C. stellifer  and  C. venustus  and an interaction between these species and EHDV hosts satisfy two of the four criteria for vector incrimination as set by the World Health Organization. Determining the vectors of EHDV is an important step towards developing sound strategies for the control of vector  Culicoides  and management of EHDV in the southeastern USA.

Field data implicating Culicoides stellifer and Culicoides venustus (Diptera: Ceratopogonidae) as vectors of epizootic hemorrhagic disease virus