Epizootic hemorrhagic disease virus in Montana: isolation and serologic survey

Epizootic hemorrhagic disease virus (EHDV) was isolated in Vero cell culture from the spleen and whole blood of a white-tailed deer (Odocoileus virginianus). A 10% spleen suspension caused acute hemorrhagic disease (HD) when inoculated into an experimental white-tailed deer and resulted in the recovery of EHDV from the blood of the experimental animal at 5 days after inoculation. The virus was identified as EHDV serotype 2 through indirect fluorescent antibody tests, electron microscopy, and reciprocal cross-neutralization tests. Approximately 73% (36/49) of the mule deer, 5% (2/42) of the white-tailed deer, and 79% (249/314) of the cattle samples tested from areas where HD had been reported were EHDV seropositive. Although none of the white-tailed deer was bluetongue virus seropositive, 29% of the mule deer and 3% of the cattle tested from "active" HD areas possessed bluetongue virus precipitating antibody.     

Light and electron microscopic comparisons of cutaneous fibromas in white-tailed and mule deer

Cutaneous fibromas of white-tailed deer (Odocoileus virginianus), when compared with normal skin of the same species, had a thinner basement membrane; thickened stratum spinosum with numerous melanocytes, desmosomes, polyribosomes, and tonofilaments; focal hyperplasia of the stratum granulosum containing numerous large, electron-dense keratohyalin granules with irregular borders and containing occasional cells with diffuse intranuclear virus particles; and a moderately thickened stratum corneum with (although rarely) small crystalline arrays of virus particles. Normal mule deer (Odocoileus hemionus) skin was structurally similar to that of the white-tailed deer. Mule deer fibromas were similar to those in white-tailed deer, except for diffuse thickening of the stratum granulosum (the cells of which contained large keratohyalin granules of various electron densities with occasional composite granules) and except for a markedly thickened stratum corneum that contained numerous intranuclear viral inclusions. In negatively stained homogenates of tumors from both deer species, viral particles resembled papillomaviruses.     

Transmission of the white-tailed deer cutaneous fibroma

Cutaneous fibromas were successfully transmitted to 7 white-tailed deer (Odocoileus virginianus) inoculated with crude fibroma extracts (2 deer) or with partially purified deer fibroma virus (5 deer). The fibromas were transmitted by intradermal and subcutaneous inoculation and by rubbing the virus preparation into tattoo sites. Inoculation by scarification was not successful. The induced tumors resembled those of naturally occurring fibromas. Tattoo inoculation sites underwent an initial acute inflammatory response followed by mesenchymal proliferation, perivascular lymphocytic infiltration, and finally regression. The deer developed antibody titers against deer fibroma virus as determined by hemagglutination inhibition, using mouse RBC. Viral antigens could not be detected by indirect immunofluorescence in any induced fibroma.     

Detection and characterisation of papillomavirus in skin lesions of giraffe and sable antelope in South Africa

Papillomavirus was detected electron microscopically in cutaneous fibropapillomas of a giraffe (Giraffa camelopardalis) and a sable antelope (Hippotragus niger). The virus particles measured 45 nm in diameter. Histopathologically, the lesions showed histopathological features similar to those of equine sarcoid as well as positive immunoperoxidase-staining of tissue sections for papillomavirus antigen. Polymerase chain reaction (PCR) detected bovine papillomavirus (BPV) DNA. Bovine papillomavirus-1 was characterised by real-time PCR in the sable and giraffe, and cloning and sequencing of the PCR product revealed a similarity to BPV-1. As in the 1st giraffe, the lesions from a 2nd giraffe revealed locally malignant pleomorphism, possibly indicating the lesional end-point of papilloma infection. Neither virus particles nor positively staining papillomavirus antigen could be demonstrated in the 2nd giraffe but papillomavirus DNA was detected by real-time PCR which corresponded with BPV-1 and BPV-2.     

Papillomavirus genomes in experimentally induced fibromas in white-tailed deer

Cutaneous fibromas of white-tailed deer were transmitted successfully to 5 young deer. Serial biopsy specimens of the induced lesions were analyzed for white-tailed deer papillomavirus, using Southern blot hybridization. Viral genomes were found in all specimens taken 1 to 7 weeks after inoculation and, in some cases, from specimens of the inoculation site obtained later. Viral DNA was found before histologic evidence of fibroblast proliferation and persisted in low copy number, compared with viral DNA of naturally developing fibromas.     

Necropsy and laboratory findings in free-living deer in South Dakota.

In a diagnostic survey of diseases in wild white-tailed deer (62 cases) and mule deer (12 cases) the most common findings were traumatic injury (20%), nontraumatic hemorrhage (13%), polioencephalomalacia (11%), and bacterial infections (9%). Although epizootic hemorrhagic disease was suspected in several cases, the virus was isolated from only 1 white-tailed deer.     

Papillomatosis of the bovine teat (mammary papilla)

A 4th of 667 cattle examined at a Wisconsin abattoir had teat papillomas. Excised teat papillomas were sorted by gross morphologic characteristics into 3 groups: (i) atypical filiform, (ii) atypical flat, and (iii) typical fibropapilloma. Bovine papilloma virus capsid antigen was detected in thin-section slides of the 3 groups of teat papillomas by peroxidase-antiperoxidase assay. The bovine papilloma virus involved with the atypical papillomas could not be characterized by molecular hybridization, because enough pure virus could not be harvested. Homogenates of the 3 groups of teat papillomas were inoculated on 2 ponies and 4 calves. Typical fibropapillomas were produced on the 4 calves, and fibromas, on the 2 ponies. Atypical papillomas were produced only in 2 heifers.     

Preliminary findings on the experimental transmission of chronic wasting disease agent of mule deer to cattle.

To determine the transmissibility of chronic wasting disease (CWD) to cattle and to provide information about clinical course, lesions, and suitability of currently used diagnostic procedures for detection of CWD in cattle, 13 calves were inoculated intracerebrally with brain suspension from mule deer naturally affected with CWD. Between 24 and 27 months postinoculation, 3 animals became recumbent and were euthanized. Gross necropsies revealed emaciation in 2 animals and a large pulmonary abscess in the third. Brains were examined for protease-resistant prion protein (PrP(res)) by immunohistochemistry and Western blotting and for scrapie-associated fibrils (SAFs) by negative-stain electron microscopy. Microscopic lesions in the brain were subtle in 2 animals and absent in the third case. However, all 3 animals were positive for PrP(res) by immunohistochemistry and Western blot, and SAFs were detected in 2 of the animals. An uninoculated control animal euthanized during the same period did not have PrP(res) in its brain. These are preliminary observations from a currently in-progress experiment. Three years after the CWD challenge, the 10 remaining inoculated cattle are alive and apparently healthy. These preliminary findings demonstrate that diagnostic techniques currently used for bovine spongiform encephalopathy (BSE) surveillance would also detect CWD in cattle should it occur naturally.     

Evaluating use of cattle winter feeding areas by elk and white-tailed deer: Implications for managing bovine tuberculosis transmission risk from the ground up

Transmission of bovine tuberculosis (Mycobacterium bovis) among wildlife and livestock has created important risks for conservation and agriculture. Management strategies aimed at controlling TB have typically been top-down, regionally focused, and government-led programs that were at best only partially successful. The purpose of this study was to quantify co-mingling of elk and white-tailed deer (WTD) with cattle at multiple spatial scales (i.e., the regional farm scale and winter cattle feeding area patch) in southwestern Manitoba, Canada, to assess the potential for bovine tuberculosis transmission and identify alternative management strategies. For each spatial scale we quantified use of cattle farms by elk and white-tailed deer. We mailed questionnaires to rural households and then conducted personal interviews with 86 cattle farmers to map the spatial distribution of their cattle winter feeding areas at a fine scale. We deployed Global Positioning System (GPS) collars on 48 wild elk and 16 wild white-tailed deer from 2003 to 2011. Elk were observed on farms by 66% of cattle producers, including 5% and 20% who observed direct and indirect contact, respectively, between elk and cattle. Cattle producers consistently (≈100%) observed white-tailed deer on their farms, including 11% and 47% whom observed direct and indirect contact, respectively, between white-tailed deer and cattle. A higher probability of white-tailed deer–cattle contact at the regional scale occurs on farms that (1) left crop residues specifically for wildlife, (2) had larger cattle herds, (3) used round bale feeders, and (4) were farther away from protected areas. None of the GPS-collared elk locations overlapped with cattle winter feeding areas. In contrast, 21% of GPS-collared white-tailed deer locations overlapped with winter cattle winter feeding areas (22% of these were from male WTD and 78% were from female WTD). White-tailed deer selected cattle winter feeding areas with higher (1) forage crop, (2) grassland/rangeland, and (3) forest cover around the cattle feeding area. Farmers overall expressed strongly negative attitudes toward eradicating the elk population or fencing the park to eradicate TB, but were generally supportive of less invasive and farm-based approaches. Our results suggested that management efforts to prevent TB transmission at the wildlife-agriculture interface can be effectively implemented using a ‘bottom-up’ approach that focuses on practical, farm-based mitigation strategies. This approach can be implemented by individual farm operators, is relatively low cost, and is generally well supported by farmers relative to other more extreme and controversial measures like wildlife eradication.     

Experimental transmission of bovine papilloma virus (BPV) extracted from morphologically distinct teat and cutaneous lesions and the effects of inoculation of BPV transformed fetal bovine cells.

Bovine papilloma virus (BPV) was extracted from five cattle each affected with only one of five morphologically distinct lesion types. When inoculated into experimental calves either by scarification or intradermal injection, the BPV extracts produced lesions macroscopically and microscopically similar to those from which individual extracts were made. Fetal bovine cells, transformed in vitro with BPV, failed to produce fibromas, fibropapillomas or papillomas when inoculated into experimental calves. When calves inoculated with virus or BPV transformed cells were challenged with the five original BPV extracts, a differential immunity was demonstrated, while control calves were susceptible to all extracts. Post mortem examination revealed the presence of upper alimentary tract papillomas in three of eight calves forming one group. These results suggest that different strains of BVP, causing morphologically separable lesion types, exist. There may be additional BPV variants causing fibropapillomas of the teat and anogenital regions of cattle. The inoculation of BPV transformed fetal bovine cells conferred a relative immunity to later challenge with some but not all BPV extracts.     

Evaluation of hunter-harvested white-tailed deer for evidence of bovine viral diarrhea virus infection in Alabama.

Bovine viral diarrhea virus (BVDV) is one of the most relevant pathogens affecting today's cattle industries. Although great strides have been made in understanding this virus in cattle, little is known about the role of wildlife in the epidemiology of BVDV. While persistently infected cattle are the most important reservoir, free-ranging ungulates may become infected with BVDV as demonstrated by serosurveys and experimental infections. Therefore, free-ranging wildlife may maintain BVDV as the result of an independent cycle and may serve as a reservoir for the virus. Systematic studies on prevalence of BVDV-specific antibodies or frequency of persistent BVDV infection in North American wildlife are sparse, and no information is available from the southeastern United States. The objective of this study was to evaluate blood and skin samples from hunter-harvested white-tailed deer (Odocoileus virginianus) for evidence of BVDV infection. Virus-neutralizing antibodies were detected in 2 of 165 serum samples. Skin biopsy immunohistochemistry (IHC) was performed on samples from 406 deer using a BVDV-specific monoclonal antibody (MAb) (15c5), and BVDV antigen was detected in one sample. A similar IHC staining pattern was obtained using a second BVDV MAb (3.12F1). Viral antigen distribution in the skin sample of this deer resembled that found in persistently infected cattle and in a previously described persistently infected white-tailed deer; thus, the deer was presumed to be persistently infected. Evidence of BVDV infection in free-ranging white-tailed deer should encourage further systematic investigation of the prevalence of BVDV in wildlife.     

Experimental persistent infection with bovine viral diarrhea virus in white-tailed deer

Bovine viral diarrhea virus (BVDV) infections cause substantial economic losses to the cattle industries. Persistently infected (PI) cattle are the most important reservoir for BVDV. White-tailed deer (Odocoileus virginianus) are the most abundant species of wild ruminants in the United States and contact between cattle and deer is common. If the outcome of fetal infection of white-tailed deer is similar to cattle, PI white-tailed deer may pose a threat to BVDV control programs. The objective of this study was to determine if experimental infection of pregnant white-tailed deer with BVDV would result in the birth of PI offspring. Nine female and one male white-tailed deer were captured and housed at a captive deer isolation facility. After natural mating had occurred, all does were inoculated intranasally at approximately 50 days of pregnancy with 10(6) CCID(50) each of a BVDV 1 (BJ) and BVDV 2 (PA131) strain. Although no clinical signs of BVDV infection were observed or abortions detected, only one pregnancy advanced to term. On day 167 post-inoculation, one doe delivered a live fawn and a mummified fetus. The fawn was translocated to an isolation facility to be hand-raised. The fawn was determined to be PI with BVDV 2 by serial virus isolation from serum and white blood cells, immunohistochemistry on skin biopsy, and RT-PCR. This is the first report of persistent infection of white-tailed deer with BVDV. Further research is needed to assess the impact of PI white-tailed deer on BVDV control programs in cattle.     

Susceptibility of cattle to first-passage intracerebral inoculation with chronic wasting disease agent from white-tailed deer

Fourteen, 3-month-old calves were intracerebrally inoculated with the agent of chronic wasting disease (CWD) from white-tailed deer (CWDwtd) to compare the clinical signs and neuropathologic findings with those of certain other transmissible spongiform encephalopathies (TSE, prion diseases) that have been shown to be experimentally transmissible to cattle (sheep scrapie, CWD of mule deer [CWDmd], bovine spongiform encephalopathy [BSE], and transmissible mink encephalopathy). Two uninoculated calves served as controls. Within 26 months postinoculation (MPI), 12 inoculated calves had lost considerable weight and eventually became recumbent. Of the 12 inoculated calves, 11 (92%) developed clinical signs. Although spongiform encephalopathy (SE) was not observed, abnormal prion protein (PrPd) was detected by immunohistochemistry (IHC) and Western blot (WB) in central nervous system tissues. The absence of SE with presence of PrPd has also been observed when other TSE agents (scrapie and CWDmd) were similarly inoculated into cattle. The IHC and WB findings suggest that the diagnostic techniques currently used to confirm BSE would detect CWDwtd in cattle, should it occur naturally. Also, the absence of SE and a distinctive IHC pattern of CWDwtd and CWDmd in cattle suggests that it should be possible to distinguish these conditions from other TSEs that have been experimentally transmitted to cattle.     

Lesions caused by Parelaphostrongylus odocoilei (Nematoda: Metastronglyloidea) in two cervid hosts

Pathologic effects and host response were evaluated in seven white-tailed deer (Odocoileus virginianus) and six mule deer (O. hemionus hemionus) each exposed per os to 300 or 1000 third-stage larvae of Parelaphostrongylus odocoilei. Pathologic effects in mule deer consisted of hemorrhagic myositis throughout skeletal muscles, severe verminous pneumonia, and moderate lymphadenitis. The major host response was a granulomatous inflammation associated with nematode eggs and larvae. Granulomas obliterated the normal architecture of affected tissues. Pathologic effects and host response were minimal in white-tailed deer. P. odocoilei is considered a potential direct or indirect pathogen in mule deer but an insignificant parasite in white-tailed deer.     

Feline cutaneous fibropapillomas: clinicopathologic findings and association with papillomavirus infection

Twenty-three feline cutaneous fibropapillomas with histologic features similar to equine sarcoids were diagnosed. They were characterized by dermal fibroblastic proliferation with overlying, often ulcerated hyperplastic epidermis. Electron microscopic findings supported the fibroblastic nature of the neoplastic cells. The 23 tumors came from 20 cats and were submitted from veterinary clinics in Wisconsin and Minnesota. These tumors occurred most commonly in young cats and were found primarily on the head, neck, and digits. Fifteen of the 17 cats for which breed was reported were domestic shorthair cats. In 11/20 cases, there was confirmed exposure to cattle. Local recurrence of the tumor following surgical excision was reported in 7 of the 18 cats for which follow-up information was available. Metastasis was not documented in any of the cases. Two of the 19 tumors tested by polymerase chain reaction (PCR) had no amplifiable DNA. The remaining 17 were positive for papillomavirus by PCR. No papillomavirus DNA was detected in three other feline skin tumors (cutaneous mast cell tumor, malignant lymphoma, and fibrosarcoma) that served as controls. This is the first report of detection of papillomavirus in feline tumors that have clinicopathologic features similar to equine sarcoids.     

EPIZOOTIOLOGY OF BLUETONGUE: THE SITUATION IN THE UNITED STATES OF AMERICA

Bluetongue was first reported in the United States in 1948 in sheep in Texas. The virus has now been isolated from sheep in 19 States. When the disease first occurs in a flock, the morbidity may reach 50 to 75% and mortality 20 to 50%. In subsequent years, the morbidity may be only 1 to 2% with very few deaths. Difference in breed susceptibility has not been observed. Natural bluetongue infection has not been observed in Angora or dairy goats. Bluetongue virus was first isolated from cattle, in Oregon, in 1959. The virus has now been isolated from cattle in 13 States. In cattle, the disease is usually inapparent but can cause mild to severe clinical disease and neonatal losses. Natural clinical bluetongue has also been reported in bighorn sheep, exotic ruminants in a zoo, mule deer, and white-tailed deer. Serological evidence of exposure to the virus has also been found in other species of ruminants in the wild. Inoculation of virulent bluetongue virus, vaccine virus, or natural disease can cause congenital deformities and neonatal losses in calves, lambs, and white-tailed deer fawns. Culicoides is considered the important insect vector of bluetongue. The virus has also been isolated from sheep keds and cattle lice. U.S. field strains of the virus fit into four serologic groups. No cross reactions were found between bluetongue and epizootic haemorrhagic disease of deer viruses. Cattle are considered significant virus reservoirs. It is necessary to use washed erythrocytes, rather than whole blood, and to inoculate susceptible sheep, rather than embryonated chicken eggs, to detect longer-term viraemia in cattle.     

Variations of response of cattle to experimentally induced viral papillomatosis.

The common bovine papilloma virus type 1 has been widely used to stimulate basic research on papilloma viruses involved in some cancers of mankind. The usually benign neoplasms of cattle caused by bovine papilloma viruses are frequent clinical problems for veterinarians. Approximately 240 experimentally induced cutaneous fibropapillomas on 8 susceptible calves had a uniform appearance of initial growth. Their size and duration ranged from about 2 months to nearly 3 years but were similar for the multiple papillomas of each calf. Sequential biopsies were done to examine the histologic changes and existence of viral antigen. Veterinarians in practice may encounter the common fibropapillomas caused by bovine papilloma virus 1 and 2 as well as papillomas with no fibromatous element. These types may develop on teats of cows in some herds. Interdigital papillomas cause a problem in some dairy herds and a virus suspected but not yet found. Prophylactic vaccination with a formalin-killed vaccine will protect against infection with bovine papilloma virus 1 and 2.     

Bovine tuberculosis in Canadian wildlife: An updated history

Mycobacterium bovis infection in wild animals attracted little attention in Canada until the disease was almost eliminated from domestic livestock. Tuberculosis was endemic in plains bison and occurred in elk, moose, and mule deer in Buffalo National Park (BNP), Alberta during the 1920s and 1930s. Bison were moved from BNP to Wood Buffalo National Park (WBNP), where tuberculosis became, and remains, endemic in bison, posing a risk to efforts to restore bison in northern Canada. Tuberculosis was found in a white-tailed deer in Ontario in 1959, and in an infected elk near Riding Mountain National Park (RMNP), Manitoba in 1992. Intense surveillance has resulted in detection of 40 elk, 8 white-tailed deer, and 7 cattle herds infected between 1997 and 2008 in the RMNP area. The strains of M. bovis in the RMNP area are different from strains tested from cattle and bison elsewhere in Canada. Management of tuberculosis in cattle and wild animals is challenging because of uncertainty about the ecology of the disease in various species, difficulty in obtaining samples and population data from wildlife, lack of validated tests, overlapping jurisdictions and authority, and conflicting values and opinions among stakeholders.     

Amplification of feline sarcoid‐associated papillomavirus DNA sequences from bovine skin

Feline sarcoids are uncommon dermal neoplasms that are thought to be caused by papillomaviral (PV) infection. Feline sarcoid‐associated PV (FeSarPV) has been consistently detected in sarcoids from North American and New Zealand cats but has not been detected within any other feline sample. This suggests that feline sarcoids may develop due to cross‐species infection by a PV from an unidentified reservoir host. While there is some epidemiological evidence to suggest that cattle are the reservoir host of FeSarPV, this PV has never been identified within any bovine sample. In this study both consensus PCR primers and primers specific to FeSarPV were used to investigate the presence of PV DNA within five fibropapillomas and 18 samples of inflammatory skin disease from cattle. Consensus primers amplified bovine PV‐2 DNA from four fibropapillomas, but none of the dermatitis samples. However, specific primers amplified FeSarPV DNA from four fibropapillomas and five inflammatory skin lesions. To the best of our knowledge this is the first time that FeSarPV has been detected within any sample other than a feline sarcoid. The ability of FeSarPV to asymptomatically infect bovine skin suggests that cattle are the reservoir host of this PV and feline sarcoids could be the result of cross‐species infection of a dead‐end host by a bovine PV.