Cross-protection between epizootic hemorrhagic disease virus serotypes 1 and 2 in white-tailed deer

Viruses in the epizootic hemorrhagic disease (EHD) serogroup are the most frequent cause of hemorrhagic disease in the southeastern United States, but nothing is known about cross-protection between the two EHD serotypes (EHDV-1 and EHDV-2) present in this region. We experimentally tested whether deer surviving EHDV-2 infection would be protected against subsequent infection with EHDV-1, and used field data to examine the possibility of reciprocal cross-protection. Eleven white-tailed deer fawns (Odocoileus virginianus) were experimentally infected with EHDV-2 and later challenged with EHDV-1. Two EHDV-2-na?ve fawns also were infected with EHDV-1. Deer were monitored via physical examination, complete blood counts, clotting profiles, viral isolation, and serology, and each animal was assigned a quantitative clinical disease severity score based on presence of certain physical and clinical parameters. Infection of na?ve controls with EHDV-1 caused severe clinical disease and death of both fawns, whereas deer previously infected with EHDV-2 exhibited no or minimal signs of disease. Thus, infection with EHDV-2 conferred protection against disease caused by subsequent EHDV-1 infection. Although prior EHDV-2 exposure protected deer from severe clinical disease, it did not prevent infection nor viremia indicating they could still act as virus amplifying hosts. These experimental infections suggest that EHDV-1 and 2 may exist in a state of mutual permissiveness.     

Epizootiology of an epizootic hemorrhagic disease outbreak in West Virginia

An outbreak of epizootic hemorrhagic disease virus, serotype 2 (EHDV-2) was responsible for localized white-tailed deer (Odocoileus virginianus) mortality in Hardy and Hampshire counties, West Virginia (USA), in the summer and fall of 1993. Using available historical data on regional herd immunity, data opportunistically collected during the epizootic, and postepizootic sampling of hunter-harvested deer, we grossly estimate certain epidemiologic parameters and compare findings to a hypothesis about hemorrhagic disease outbreaks in the Appalachian Mountains. During the epizootic, 57.9 km(2) were actively searched and 228 dead deer were found. Epizootic hemorrhagic disease virus, serotype 2 was isolated from seven of nine deer sampled in Hardy and Hampshire counties. Preepizootic exposure of deer to EHD viruses was unknown, but available data suggest that it was negligible. The geographic distribution of the outbreak was defined by plotting the locations of dead deer found during the outbreak, as well as the locations of deer harvested by hunters after the outbreak that had antibodies to EHDV-2 on a map sectioned into 16.65 km(2) rectangular sections. Sections that included one or more dead deer or hunter-harvested deer with antibodies to EHDV-2 were included in the defined outbreak area. Postoutbreak sampling revealed monospecific EHDV-2 antibodies in 12% of deer harvested by hunters within the defined outbreak area. Based on the available data and accepting certain assumptions, gross calculations suggest that this outbreak appears to have been isolated and probably killed a high percentage of the deer that were infected. This is consistent with the hypothesis that sporadic hemorrhagic disease outbreaks in the Appalachian Mountains are usually localized and severe.     

Innate resistance to epizootic hemorrhagic disease in white-tailed deer

Differences in innate disease resistance at the sub-species level have major implications for wildlife management. Two subspecies of white-tailed deer, Odocoileus virginianus borealis and O. virginianus texanus were infected with epizootic hemorrhagic disease (EHD) viruses. These viruses are highly virulent pathogens of white-tailed deer and are endemic within the range of O. virginianus texanus but not within the range of O. virginianus borealis. Two experimental infections were performed. Five O. virginianus texanus fawns and five O. virginianus borealis fawns were infected with 10(7.1) median tissue culture infective doses (TCID50) of EHD virus, serotype 1 and five of each subspecies were infected with 10(7.1) TCID50 of EHD virus, serotype 2. Infections with both EHD virus serotypes caused severe clinical disease and mortality in O. virginianus borealis fawns, whereas disease was mild or nondetectable in O. virginianus texanus fawns. Virus titers and humoral immune response were similar in both subspecies suggesting that differences in innate disease resistance explain the differences seen in clinical disease severity. In white-tailed deer, innate disease resistance may vary at the subspecies level. Should this phenomenon occur in other species, these findings have major implications for managing wildlife populations, both endangered and non-endangered, using tools such as translocation and captive propagation.     

Molecular characterization of epizootic hemorrhagic disease virus serotype 1 associated with a 1999 epizootic in white-tailed deer in the eastern United States

During the autumn of 1999 (mid-August-late September), an outbreak of hemorrhagic disease in white-tailed deer (Odocoileus virginianus) caused by epizootic hemorrhagic disease virus serotype 1 (EHDV-1) occurred along the east coast of the United States from Georgia to New Jersey. An EHDV-1 epizootic of such magnitude had not been described in this region since 1975. To determine the genetic relatedness among the 1999 viruses, as well as among additional EHDV-1 isolates from the eastern and western United States, portions of the S10 and L2 gene segments were sequenced and compared utilizing phylogenetic analyses. Nearly all of the 1999 eastern isolates were identical in nucleotide sequence at one or both loci. Additionally, confirmed cases of EHDV-1 in white-tailed deer occurred in a south (Georgia)-to-north (New Jersey/Virginia) progression over a short period of approximately six weeks. Taken together, these results indicate that this outbreak resulted from the spread of a single viral strain. The phylograms derived from analysis of the entire sample set displayed eastern and western region-specific clusterings (topotypes), as well as an eastern versus western difference in branch lengths, which may reflect the influence of epizootic versus enzootic transmission patterns on viral genetic diversity.     

Dynamics of maternal antibodies to hemorrhagic disease viruses (Reoviridae: Orbivirus) in white-tailed deer

Enzootic stability, potentially associated with acquired resistance and subsequent transfer of maternal antibodies, innate resistance, or both, has been hypothesized to explain the lack of reports of hemorrhagic disease (HD) in white-tailed deer (Odocoileus virginianus) from Texas. The objectives of this research were to determine the following: how long maternal antibodies to epizootic hemorrhagic disease (EHD) and bluetongue (BT) viruses persist; whether fawns from an enzootic site are naturally exposed to EHD and BT viruses while maternal antibodies are present; and whether field-challenged fawns develop clinical disease. Twelve of 52 fawns from Texas were moved to an indoor facility. All 12 (100%) were positive for maternal antibodies to EHD or BT viruses by agar gel immunodiffusion (AGID) and serum neutralization (SN) tests. Weekly monitoring demonstrated that precipitating antibodies disappeared by 23 wk of age and serum neutralizing antibodies disappeared by 17-18 wk of age. Fawns that remained outdoors in Texas were not observed with signs of HD. At 14-21 wk of age (October), 39 of 40 (98%) fawns that had remained outdoors were positive for EHD and/or BT virus antibodies by AGID and 32 (80%) had SN antibody titers to one or more of five viruses (EHDV-1, EHDV-2, BTV-10, BTV-11, BTV-17). Antibody titers to EHDV-1, EHDV-2, and BTV-11 all exceeded titers of same-age indoor fawns, suggesting recent exposure. Epizootic hemorrhagic disease viruses were isolated from seven (18%) of the outdoor fawns and all 40 remained clinically normal. Natural exposure of deer to EHD and BT viruses occurred at this site in the presence of maternal antibodies without causing disease. This may be due to acquired immunity and the subsequent transfer of maternal antibodies, but it does not exclude innate resistance as a possible factor in the enzootic stability of EHD and BT viruses at this location.     

Development of a model of natural infection with Mycobacterium bovis in white-tailed deer.

The objective of this study was to develop a suitable experimental model of natural Mycobacterium bovis infection in white-tailed deer (Odocoileus virginianus), describe the distribution and character of tuberculous lesions, and to examine possible routes of disease transmission. In October 1997, 10 mature female white-tailed deer were inoculated by intratonsilar instillation of 2 x 10(3) (low dose) or 2 x 10(5) (high dose) colony forming units (CFU) of M. bovis. In January 1998, deer were euthanatized, examined, and tissues were collected 84 to 87 days post inoculation. Possible routes of disease transmission were evaluated by culture of nasal, oral, tonsilar, and rectal swabs at various times during the study. Gross and microscopic lesions consistent with tuberculosis were most commonly seen in medial retropharyngeal lymph nodes and lung in both dosage groups. Other tissues containing tuberculous lesions included tonsil, trachea, liver, and kidney as well as lateral retropharyngeal, mandibular, parotid, tracheobronchial, mediastinal, hepatic, mesenteric, superficial cervical, and iliac lymph nodes. Mycobacterium bovis was isolated from tonsilar swabs from 8 of 9 deer from both dosage groups at least once 14 to 87 days after inoculation. Mycobacterium bovis was isolated from oral swabs 63 and 80 days after inoculation from one of three deer in the low dose group and none of four deer in the high dose group. Similarly, M. bovis was isolated from nasal swabs 80 and 85 days after inoculation in one of three deer from the low dose group and 63 and 80 days after inoculation from two of four deer in the high dose group. Intratonsilar inoculation with M. bovis results in lesions similar to those seen in naturally infected white-tailed deer; therefore, it represents a suitable model of natural infection. These results also indicate that M. bovis persists in tonsilar crypts for prolonged periods and can be shed in saliva and nasal secretions. These infected fluids represent a likely route of disease transmission to other animals or humans.     

Hemorrhagic disease in deer in Arizona

Two mule deer (Odocoileus hemionus) and one white-tailed deer (Odocoileus virginianus) in Arizona (USA) were submitted for necropsy. Gross and microscopic lesions compatible with hemorrhagic disease (HD) were observed in all three deer. Epizootic hemorrhagic disease virus type 2 (EHDV-2) was isolated from two of the deer. To our knowledge, this is the first documentation of HD in deer in Arizona. Two of the mortalities were attributed to EHDV-2 infection.     

Review of the 2012 Epizootic Hemorrhagic Disease Outbreak in Domestic Ruminants in the United States

An unusually large number of cases of Epizootic hemorrhagic disease (EHD) were observed in United States cattle and white-tailed deer in the summer and fall of 2012. USDA APHIS Veterinary Services area offices were asked to report on foreign animal disease investigations and state diagnostic laboratory submissions which resulted in a diagnosis of EHD based on positive PCR results. EHD was reported in the following species: cattle (129 herds), captive white-tailed deer (65 herds), bison (8 herds), yak (6 herds), elk (1 herd), and sheep (1 flock). A majority of the cases in cattle and bison were found in Nebraska, South Dakota, and Iowa. The majority of cases in captive white-tailed deer were found in Ohio, Iowa, Michigan, and Missouri. The most common clinical sign observed in the cattle and bison herds was oral lesions. The major observation in captive white-tailed deer herds was death. Average within-herd morbidity was 7% in cattle and bison herds, and 46% in captive white-tailed deer herds. The average within-herd mortality in captive white-tailed deer herds was 42%.     

Experimental and field studies of Escherichia coli O157:H7 in white-tailed deer

Studies were conducted to evaluate fecal shedding of Escherichia coli O157:H7 in a small group of inoculated deer, determine the prevalence of the bacterium in free-ranging white-tailed deer, and elucidate relationships between E. coli O157:H7 in wild deer and domestic cattle at the same site. Six young, white-tailed deer were orally administered 10(8) CFU of E. coli O157:H7. Inoculated deer were shedding E. coli O157:H7 by 1 day postinoculation (DPI) and continued to shed decreasing numbers of the bacteria throughout the 26-day trial. Horizontal transmission to an uninoculated deer was demonstrated. Although E. coli O157:H7 bacteria were recovered from the gastrointestinal tracts of deer necropsied from 4 to 26 DPI, attaching and effacing lesions were not apparent in any deer. Results are similar to those of inoculation studies in calves and sheep. In field studies, E. coli O157 was not detected in 310 fresh deer fecal samples collected from the ground. It was detected in feces, but not in meat, from 3 of 469 free-ranging deer in 1997. In 1998, E. coli O157 was not detected in 140 deer at the single positive site found in 1997; however, it was recovered from 13 of 305 dairy and beef cattle at the same location. Isolates of E. coli O157:H7 from deer and cattle at this site differed with respect to pulsed-field gel electrophoresis patterns and genes encoding Shiga toxins. The low overall prevalence of E. coli O157:H7 and the identification of only one site with positive deer suggest that wild deer are not a major reservoir of E. coli O157:H7 in the southeastern United States. However, there may be individual locations where deer sporadically harbor the bacterium, and venison should be handled with the same precautions recommended for beef, pork, and poultry.     

Experimental infection of white-tailed deer (Odocoileus virginianus) with Mycobacterium avium subsp. paratuberculosis

Mycobacterium avium subsp. paratuberculosis (Map) is the causative agent of paratuberculosis or Johne's disease, a chronic enteric disease of domestic ruminants as well as some nondomestic ruminants. Paratuberculosis is characterized by a protracted subclinical phase followed by clinical signs such as diarrhea, weight loss, and hypoproteinemia. Fecal shedding of Map is characteristic of both the subclinical and clinical phases, and it is important in disease transmission. Lesions of paratuberculosis are characterized by chronic granulomatous enteritis and mesenteric lymphadenitis. Animal models of paratuberculosis that simulate all aspects of the disease are rare. Oral inoculation of 9-day-old white-tailed deer (Odocoileus virginianus) on 3 June 2002 with 1.87 x 10(10) colony-forming units of Map strain K10 resulted in clinical disease (soft to diarrheic feces) as early as 146 days after inoculation; lesions consistent with paratuberculosis were observed in animals at the termination of the study. Intermittent fecal shedding of Map was seen between 28 and 595 days (4 March 2004) after inoculation. These findings suggest that experimental oral inoculation of white-tailed deer fawns may mimic all aspects of subclinical and clinical paratuberculosis.     

Parelaphostrongylus andersoni (Nematoda: Protostrongylidae) in white-tailed deer from Michigan

Dorsal-spined larvae in fecal samples from free-ranging white-tailed deer (Odocoileus virginianus) in Michigan and Pennsylvania were used as a source of larvae to infect a hand-raised white-tailed deer fawn. The fawn receive 200 third-stage larvae and passed dorsal-spined larvae in feces 66 days later. Muscleworm (Parelaphostrongylus andersoni), and meningeal worm (Parelaphostrongylus tenuis) were recovered at necropsy. Two white-tailed deer and seven wapiti (Cervus elaphus) exposed to larvae of the source from Pennsylvania harbored only P. tenuis. This is the first report of P. andersoni in the midwestern United States and extends the known range of this muscleworm in free-ranging white-tailed deer. Concurrent infections of P. andersoni and P. tenuis have not been established previously in experimentally infected fawns.     

Susceptibility of white-tailed deer (Odocoileus virginianus) to experimental infection with epizootic hemorrhagic disease virus serotype 7

During the fall of 2006, in Israel, epizootic hemorrhagic disease virus (EHDV) serotype 7 caused an intense and widespread epizootic in domestic cattle that resulted in significant economic losses for the dairy industry. The susceptibility of potential North American vector and ruminant hosts to infection with EHDV-7 is not known but is essential to understanding the potential for establishment of this exotic orbivirus in North America if it were introduced. Our primary objective was to determine whether white-tailed deer (WTD; Odocoileus virginianus) are susceptible to infection with EHDV-7. Six, 8-mo-old WTD were experimentally infected with EHDV-7, and all became infected and exhibited varying degrees of clinical disease. Clinical signs, clinicopathologic abnormalities, and postmortem findings were consistent with previous reports of orbiviral hemorrhagic disease (HD) in this species. Four of six animals died or were euthanized because of the severity of disease, one on postinoculation day (PID) 5 and the remaining WTD on PID 7. All deer had detectable viremia on PID 3, which peaked on PID 5 or 6 and persisted for as long as PID 46 in one animal. Deer surviving the acute phase of the disease seroconverted by PID 10. Based on the 67% mortality rate we observed, this strain of EHDV-7 is virulent in WTD, reaffirming their role as a sentinel species for the detection of endemic and nonendemic EHDV. Further, the observed disease was indistinguishable from previous reports of disease caused by North American EHDV and bluetongue virus serotypes, highlighting the importance of serotype-specific diagnostics during suspected HD outbreaks.     

Parasites, diseases, and health status of sympatric populations of fallow deer and white-tailed deer in Kentucky

In August 1983, a study on parasites, diseases, and health status was conducted on sympatric populations of fallow deer (Dama dama) and white-tailed deer (Odocoileus virginianus) from Land Between The Lakes, Lyon and Trigg counties, Kentucky. Five adult deer of each species were studied. White-tailed deer had antibodies to epizootic hemorrhagic disease (EHD) virus and Leptospira interogans serovariety icterohemorrhagiae, and fallow deer had antibodies to bluetongue and EHD viruses. Serologic tests for bovine virus diarrhea virus, infectious bovine rhinotracheitis virus, parainfluenza3 virus, and Brucella spp. were negative. One white-tailed deer had an infectious cutaneous fibroma, and one fallow deer had pulmonary mucormycosis. White-tailed deer harbored 16 species of parasites, all of which are considered typical of the parasite fauna of this host in the southeastern United States. Fallow deer harbored nine species of parasites, including eight species known to occur in white-tailed deer on the area and one species (Spiculopteragia assymmetrica) that is not. All fallow deer had inflammatory lesions in the spinal cord and/or brain that were attributed to prior infection with meningeal worm (Parelaphostrongylus tenuis), indicating that P. tenuis infections are not always fatal for this species. The apparent high rate of exposure of Land Between The Lakes fallow deer to P. tenuis without a resultant high rate of clinical cerebrospinal parelaphostrongylosis is hypothesized to be due to a low prevalence and intensity of P. tenuis, partial innate resistance of fallow deer, and acquired immunity.     

Experimental and Field Studies of Escherichia coli O157:H7 in White-Tailed Deer

Studies were conducted to evaluate fecal shedding of Escherichia coli O157:H7 in a small group of inoculated deer, determine the prevalence of the bacterium in free-ranging white-tailed deer, and elucidate relationships between E. coli O157:H7 in wild deer and domestic cattle at the same site. Six young, white-tailed deer were orally administered 10⁸ CFU of E. coli O157:H7. Inoculated deer were shedding E. coli O157:H7 by 1 day postinoculation (DPI) and continued to shed decreasing numbers of the bacteria throughout the 26-day trial. Horizontal transmission to an uninoculated deer was demonstrated. Although E. coli O157:H7 bacteria were recovered from the gastrointestinal tracts of deer necropsied from 4 to 26 DPI, attaching and effacing lesions were not apparent in any deer. Results are similar to those of inoculation studies in calves and sheep. In field studies, E. coli O157 was not detected in 310 fresh deer fecal samples collected from the ground. It was detected in feces, but not in meat, from 3 of 469 free-ranging deer in 1997. In 1998, E. coli O157 was not detected in 140 deer at the single positive site found in 1997; however, it was recovered from 13 of 305 dairy and beef cattle at the same location. Isolates of E. coli O157:H7 from deer and cattle at this site differed with respect to pulsed-field gel electrophoresis patterns and genes encoding Shiga toxins. The low overall prevalence of E. coli O157:H7 and the identification of only one site with positive deer suggest that wild deer are not a major reservoir of E. coli O157:H7 in the southeastern United States. However, there may be individual locations where deer sporadically harbor the bacterium, and venison should be handled with the same precautions recommended for beef, pork, and poultry.     

Culicoides, the vector of epizootic hemorrhagic disease in white-tailed deer in Kentucky in 1971.

The biting gnat, Culicoides variipennis (Coquillett), was shown to be a vector of epizootic hemorrhagic disease (EHD) in white-tailed deer, Odocoileus virginianus, in Kentucky because of virus isolations from parous females. Epidemiological evidence showed a close relationship of this vector to the animal host during an outbreak of EHD in penned deer. Larval breeding sites of C. variipennis were found and C. variipennis was the most abundant biting fly present during the outbreak. Females of C. variipennis were commonly observed biting deer, swine, cattle and, occasionally, man.     

Antibodies to bluetongue and epizootic hemorrhagic disease viruses in a barrier island white-tailed deer population.

From 1981 through 1989, serum samples from 855 white-tailed deer (Odocoileus virginianus) from Ossabaw Island, Georgia (USA), were tested for antibodies to bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV). During this period, prevalence of precipitating antibodies to BTV and EHDV as determined by agar gel immunodiffusion (AGID) tests decreased from 74% to 3% and from 34% to 1%, respectively. Antibodies were detected in serum samples from 0.5-yr-old deer only during 1981, 1982, and 1983, and with few exceptions, positive serological results after 1983 were restricted to older age classes. A decrease in prevalence of precipitating antibodies to BTV and EHDV in age classes exposed during 1981 indicates that AGID results from white-tailed deer populations underestimate the extent of previous exposure to these viruses. Serum neutralization test results from AGID-positive deer indicated that BTV 11 was the principal serotype responsible for infections during 1981. Since 1983, this serotype has been replaced by BTV 13; however, there has been a low level of transmission within the herd. Infection with EHDV 2 appeared most prevalent during 1982; as with BTV 13, there has been limited transmission in this high density deer population since 1983.     

The potential for false-positive diagnosis of protostrongyliasis by extraction of larvae from feces

The potential of protostrongylid first-stage larvae (L1) to survive passage through the alimentary canal of non-infected mammals was investigated. Parelaphostrongylus tenuis L1 were collected from feces of an experimentally infected white-tailed deer (Odocoileus virginianus). We utilized two red deer (Cervus elaphus) and four laboratory rats (Rattus norvegicus) which were each fed the L1 of P. tenuis. Larvae were recovered, intact and alive, from the fecal samples of all six animals. Larvae of P. tenuis, and probably of other related species, can survive passage through the alimentary canal of uninfected mammals and they can be collected from feces using the Baermann technique and other related larval extraction methods. Rain water was found to be successful in the dispersal of P. tenuis L1 from the feces of infected animals. These findings raise the possibility of ingestion of L1 and their subsequent passage, by uninfected animals. This potential for false-positive diagnosis of infection in live animals necessitates accurate interpretation of a host's infection-status. Such findings reinforce the need for a reliable method of diagnosing infections in live animals.     

Aerosol exposure of white-tailed deer (Odocoileus virginianus) to Mycobacterium bovis

Tuberculosis due to Mycobacterium bovis affects both captive and free-ranging Cervidae in the United States. Various animal models have been developed to study tuberculosis of both humans and animals. Generally, tuberculosis is transmitted by aerosol and oral routes. Models of aerosol exposure of large animals to M. bovis are uncommon. In order to develop a reliable method of aerosol exposure of white-tailed deer (Odocoileus virginianus) to M. bovis, 12 healthy white-tailed deer, aged 8-10 mo, were infected by aerosol exposure to 2 x 10(5) to 1 x 10(6) colony forming units (CFU) (high close, n = 4) of M. bovis or 6 x 10(2) to 1.6 x 10(3) CFU (low dose, n = 8) of M. bovis. Tuberculous lesions were more widely disseminated in (leer receiving the high dose, while lesions in deer receiving the low dose were more focused on the lungs and associated lymph nodes (tracheobronchial and mediastinal). Aerosol delivery of M. bovis to white-tailed deer results in a reliable manner of experimental infection that may be useful for studies of disease pathogenesis, immune response, mycobacterial shedding, and vaccine efficacy.