EVIDENCE OF EPIZOOTIC HEMORRHAGIC DISEASE VIRUS AND BLUETONGUE VIRUS EXPOSURE IN NONNATIVE RUMINANT SPECIES IN NORTHERN FLORIDA.

Epizootic hemorrhagic disease virus (EHDV) and bluetongue virus (BTV) are vector-borne viruses of ruminants nearly worldwide. They can affect white-tailed deer (WTD; Odocoileus virginianus), the ranching industry, and nonindigenous hoof stock species managed for conservation. One potential risk factor for ranched WTD is commingling with nonindigenous species on high-fenced properties. Nonindigenous species provide novel viewing and hunting opportunities; however, their presence may create disease hazards. Furthermore, animals within conservation properties may be at a risk from commingling exotics and adjacent wild WTD. Currently, knowledge about EHDV and BTV seroprevalence and transmission is limited in nonindigenous populations in the southeastern United States. The authors conducted a serological survey of 10 Bovidae and 5 Cervidae species residing within two properties in northern Florida. The first site was a conservation property breeding threatened nonindigenous species for conservation. The second property was a private high-fenced game preserve managing WTD and nonindigenous species for breeding, sale, and harvest. Blood samples were tested for titers to three EHDV serotypes (1, 2, and 6) and active circulating viral EHDV and BTV. The private ranch had evidence of EHDV or BTV in one of three (33.3%) Bovidae species and four of five (80%) Cervidae species sampled. At the conservation property, evidence of EHDV infection was found in four of seven (57.1%) Bovidae and one of one (100%) Cervidae species sampled. The presence of antibodies in many nonindigenous species sampled might indicate these species are potential viral hosts and may be a risk to ranched WTD and other species within the same property. Nonindigenous species within the private ranch and conservation properties are at risk of contracting EHDV and BTV, and herd managers should reduce vector-host interactions and consider increased biosecurity measures when translocating animals.

Epizootic Hemorrhagic Disease Virus and Bluetongue Virus Seroprevalence in Wild White-Tailed Deer (Odocoileus virginianus) in Florida, USA.

A wild population of white-tailed deer (Odocoileus virginianus) was surveyed for evidence of past or current epizootic hemorrhagic disease virus (EHDV) and current bluetongue virus (BTV) infections. We collected 121 blood samples from hunter-harvested or live-captured deer from two state-managed properties in northwest Florida, US; live captures were in support of a movement ecology study. Blood samples were tested for antibodies against titers to three EHDV serotypes (EHDV-1, EHDV-2, and EHDV-6), and multiplex quantitative reverse transcription PCR was used to identify the presence of EHDV or BTV viral RNA. Of these samples, 81% (98/121) tested seropositive for at least one of three serotypes of EHDV. Of those testing seropositive, 33% (40/121) contained antibodies for two serotypes, and 19% (24/121) contained antibodies for all three EHDV serotypes. Furthermore, results of generalized linear models indicated that the probability of infection by EHDV serotypes 1 and 6 increased with an animal's age. Our findings indicate that seroprevalence may be high for multiple serotypes in regions where these orbiviruses are endemic. These results could prove useful for managing disease risk in naïve deer populations.

Three New Orbivirus Species Isolated from Farmed White-Tailed Deer (Odocoileus virginianus) in the United States.

We report the detection and gene coding sequences of three novel Orbivirus species found in six dead farmed white-tailed deer in the United States. Phylogenetic analyses indicate that the new orbiviruses are genetically closely related to the Guangxi, Mobuck, Peruvian horse sickness, and Yunnan orbiviruses, which are thought to be solely borne by mosquitos. However, four of the six viruses analyzed in this work were found as co-infecting agents along with a known cervid pathogen, epizootic hemorrhagic disease virus-2 (EHDV-2), raising questions as to whether the new viruses are primary pathogens or secondary pathogens that exacerbate EHDV-2 infections. Moreover, EHDV-2 is known to be a Culicoides-borne virus, raising additional questions as to whether Culicoides species can also serve as vectors for the novel orbiviruses, if mosquitoes can vector EHDV-2, or whether the deer were infected through separate bites by the insects. Our findings expand knowledge of the possible viral pathogens of deer in the United States. Moreover, due to the close genetic relatedness of the three new orbiviruses to viruses that are primary pathogens of cattle and horses, our findings also underscore a crucial need for additional research on the potential role of the three new orbiviruses as pathogens of other animals.

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

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.