PCR detection of bovine herpesviruses from nonbovine ruminants in Hungary

Polymerase chain reaction (PCR) was used to test six different nonbovine ruminant species for five bovine herpesviruses including infectious bovine rhinotracheitis virus (BoHV-1), bovine herpes mammillitis virus (BoHV-2), Movar-type herpesvirus (BoHV-4), bovine herpesvirus type 5 (BoHV-5), and alcelaphine herpesvirus 1 (AlHV-1). Species tested included 56 roe deer (Capreolus capreolus), 66 red deer (Cervus elaphus), 20 fallow deer (Dama dama), 16 mouflon (Ovis musimon), 34 domestic sheep, and 44 domestic goats, which were sampled in Hungary in 2003. Tracheal and popliteal lymph nodes collected from these animals were tested for the presence of the five bovine herpesviruses using three nested (two of which were duplex) PCR assays. Three bovine herpesviruses (BoHV-1, -2, and -4) were detected, whereas no evidence of AlHV-1 or BoHV-5 was observed. Prevalence of BoHV-1 ranged from 12% to 47%, and PCR-positive results were observed in all species tested. BoHV-2 was detected from roe deer, red deer, fallow deer, mouflon, and domestic sheep, and the prevalence in these species ranged from 3% to 50%. BoHV-4 was detected in all species, with prevalence ranging from 12% to 69%. Sequenced PCR products were 99-100% identical to bovine herpesviral sequences deposited in the GenBank.     

Serologic survey of white-tailed deer on Anticosti Island, Quebec for bovine herpesvirus 1, bovine viral diarrhea, and parainfluenza 3.

In 1985 unusual mortality was observed among the 3- to 4-yr-old white-tailed deer (Odocoileus virginianus) on Anticosti Island, Québec (Canada). A viral pathogen was suspected to be the cause of the deaths. Thus, a serologic survey for bovine herpesvirus 1 (BHV-1), bovine viral diarrhea (BVD) virus and parainfluenza-3 (PI-3) virus was conducted. We examined 396 deer sera from 1985. Results indicated that the high mortality mainly afflicted 3- to 4-yr-old deer. In 1985, 57% of deer sampled were seropositive for viral neutralizing antibodies against BHV-1. Prevalences decreased over the next 2 yr of the survey. Prevalence of antibodies against PI-3 virus, determined by hemagglutination inhibition test, remained high (82% to 84%) for the 3 yr period. No deer were seropositive for neutralizing antibodies against BVD virus during the survey period. Analysis of antibodies against BHV-1 and PI-3 viruses according to sex, age and antibody titers revealed that an epizootic BHV-1 infection occurred in 1985; PI-3 infection appears to be enzootic in Anticosti deer.     

Antibodies to ruminant alpha-herpesviruses and pestiviruses in Norwegian cervids

A serologic survey revealed that Norwegian populations of free-ranging reindeer (Rangifer tarandus tarandus), roe deer (Capreolus capreolus), red deer (Cervus elaphus), and moose (Alces alces) have been exposed to alpha-herpesviruses and pestiviruses. A total of 3,796 serum samples collected during the period 1993-2000 were tested in a neutralization test for antibodies against bovine herpesvirus 1 (BHV-1) or cervid herpesvirus 2 (CerHV-2), and 3,897 samples were tested by a neutralization test and/or enzyme-linked immunosorbent assay for antibodies against bovine viral diarrhea virus (BVDV). Antibodies against alpha-herpesvirus were found in 28.5% of reindeer, 3.0% of roe deer, and 0.5% of red deer, while all moose samples were negative. In reindeer, the prevalence of seropositive animals increased with age and was higher in males than females. Antibodies against BVDV were detected in 12.3% of roe deer, 4.2% of reindeer, 2.0% of moose and 1.1% of red deer. The results indicate that both alpha-herpesvirus and pestivirus are endemic in reindeer and pestivirus is endemic in roe deer in Norway. The viruses may be specific cervid strains. Seropositive red deer and moose may have become exposed as a result of contact with other ruminant species.     

Response of white-tailed deer to infection with peste des petits ruminants virus

White-tailed deer (Odocoileus virginianus) were infected experimentally with two strains of peste des petits ruminants virus. The response varied from fatal consequence to subclinical infection. The clinical signs and gross lesions were similar to those in goats. Virus was recovered from all the infected deer, and survivors developed specific antibodies demonstrated by complement fixation and virus neutralization tests. Survivors also resisted challenge with virulent rinderpest virus that was lethal to a control deer.     

Serological evidence of bovine herpesvirus 1-related virus infection in the white-tailed deer population on Anticosti Island, Quebec

High white-tailed deer (Odocoileus virginianus) population densities and the occurrence of harsh environmental conditions are present on Anticosti Island, located in the Gulf of Saint-Lawrence (Quebec, Canada). This island is the northernmost region of white-tailed deer distribution in northeastern North America. The aim of this work was to determine whether a herpesvirus serologically related to bovine herpesvirus 1 (BHV1) may occur in a stressed white-tailed deer population. One hundred one deer sera were collected from apparently healthy animals during the hunting season from September to late November 1985. Fifty-three percent of tested deer were positive to the seroneutralization test using Colorado strain of BHV1 virus. Higher percentages of seropositivity were observed in animals of both sexes greater than 4-yr-old. Analysis of antibody titers in seropositive animals according to age suggests that BHV1-related viral infection is endemic in the Anticosti Island deer population. It is postulated that environmental stress may induce immunosuppression of certain infected and/or carrier animals in their population that shed virus for long periods of time.     

Prevalence of chronic wasting disease and bovine tuberculosis in free-ranging deer and elk in South Dakota

Heads of hunter-harvested deer and elk were collected throughout South Dakota (USA) and within established chronic wasting disease (CWD) surveillance areas from 1997-2002 to determine infection with CWD and bovine tuberculosis (TB). We used immunohistochemistry to detect CWD-infected individuals among 1,672 deer and elk sampled via geographically targeted surveillance. A total of 537 elk (Cervus elaphus nelsoni), 813 white-tailed deer (Odocoileus virginianus), and 322 mule deer (O. hemionus) was sampled for CWD. Estimated overall prevalence and associated confidence intervals (95%) in white-tailed deer was 0.001% (0-0.007%). Similarly, estimated overall prevalence in elk and mule deer was 0.0% (0-0.004%) and 0.0% (0-0.011%), respectively. A total of 401 elk, 1,638 white-tailed deer, and 207 mule deer was sampled for TB. Estimated overall prevalence of infection with TB in elk harvested in South Dakota was 0.0% (0-0.009%). Similarly, estimated overall prevalence of TB in white-tailed deer and mule deer harvested throughout South Dakota was 0.0% (0-0.002%) and 0.0% (0-0.018%), respectively.     

Survey for vesicular stomatitis virus neutralizing antibodies in serums of white-tailed deer Odocoileus virginianus of the southeastern United States

New Jersey type vesicular stomatitis (VS) antibodies were found in 14 of 677 deer serums tested by neutralization tests in embryonated chicken eggs. Twelve positive serums were received from Louisiana and two from Georgia. Eight of the positive deer serums from Louisiana were collected in the area of the only reported case of VS during 1967. Clinical VS has not been diagnosed in the east coast states since 1964. Two positive deer serums were collected on Ossabaw Island, Georgia, and three positive serums, one each from a hog, bull, and sheep, were collected from young animals on the island. These findings indicated subclinical infection or a nidus of New Jersey VS in Georgia. The low percentage of New Jersey type VS antibodies in deer and the distribution parallel the low incidence of VS since 1964. No antibodies were present for Indiana type VS virus.     

Bluetongue virus: (1) in pregnant white-tailed deer (2) a plaque reduction neutralization test

Six white-tailed deer (Odocoileus virginianus) were infected with bluetongue virus (BTs, vaccinal strain) approximately one-third of the way through their gestation period. One deer died of bluetongue 21 days after inoculation. Of the five surviving the infection, one had two mummified fetuses, and the others no fetuses upon euthanasia two weeks after term. Fetuses were present in two control deer and in the one which died of bluetongue. A plaque reduction neutralization test for bluetongue virus was developed and described for the first time and its sensitivity illustrated by high post inoculation titers which ranged from 1:3200 to greater than 1:16000.     

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.     

Occurrence of antibodies to the etiologic agents of infectious bovine rhinotracheitis, parainfluenza 3, leptospirosis, and brucellosis in white-tailed deer in Minnesota

A serologic survey was conducted on 628 white-tailed deer (Odocoileus virginianus) in 1976 and 1979-1980. Tests for antibodies to the etiologic agents of infectious bovine rhinotrancheitis (IBR), parainfluenza 3 (PI3), leptospirosis, and brucellosis produced positive results of 15%, 20%, 3% and 0%, respectively. Adult deer had significantly higher prevalence of antibodies to IBR virus and PI3 virus than fawns. These data provide a basis for monitoring these disease agents in Minnesota's white-tailed deer.     

Herpesvirus infection in woodland caribou in Alberta, Canada

Sera and genital swabs collected from 121 adult woodland caribou (Rangifer tarandus caribou) in five subpopulations in northern Alberta, Canada, between December 1997 and October 1999, were examined for evidence of infection with herpesviruses or pestiviruses. No virus was isolated from sera or swabs, and no antibodies against bovine viral diarrhea virus were detected. However, 63 (52%) of the 121 animals had neutralizing antibody titers against bovine herpesvirus 1. There was sufficient serum from 37 of the 121 caribou to allow parallel testing for antibodies against a new alphaherpesvirus isolated from an elk (Cervus elaphus nelsoni), and 20 animals had antibodies against this virus. Paired sera collected 11 mo apart from 14 caribou showed seroconversion in seven animals, indicating that an active herpesvirus infection was present. Virus neutralization data suggest that these caribou are infected with a distinct alphaherpesvirus.     

An epizootic of hemorrhagic disease in white-tailed deer in Missouri

As part of a white-tailed deer (Odocoileus virginianus) survival study in Missouri (USA) we were actively monitoring 97 radio-collared deer when 8 (8%) died. This mortality, which occurred from 20 August to 23 September 1996, consisted of five adult females, two yearling females and one yearling male. Based on the seasonality of this mortality and the isolation of epizootic hemorrhagic disease virus (EHDV) serotype 2 from one of these animals, we believe that these losses resulted from an epizootic of hemorrhagic disease. The remains of five unmarked deer that may have died from HD also were found on the study area during this same period. During the fall following this mortality, we tested serum from 96 deer taken by hunters in the immediate area. Fifteen (16%) were positive for EHDV or bluetongue virus (BTV) antibodies as determined by agar gel immunodiffusion tests. Serum neutralization test results indicated that previous infections were caused by EHDV virus serotype 2. Based on these data, and assuming that there was no prior exposure to EHDV serotype 2 in this population, the exposure rate for this epizootic was 24% of which 8% died. We noted hoof interruptions in only two of the 96 deer sampled. During this mortality event, the Missouri Department of Conservation received no reports of dead deer, and without the radio-monitored animals the event would have been undetected.     

Persistence of Escherichia coli introduced into streambed sediments with goose,deer and bovine animal waste

Aims:  The focus of this work was to compare the survival of Escherichia coli introduced into streambed sediments from goose, deer and bovine faeces vs indigenous E. coli. Methods and Results:  The survival experiments were conducted in flow‐through chambers for 32 days using two sediments (mineral and organic) obtained from a first‐order creek in Maryland. Bovine, goose and deer faeces were collected fresh and diluted or enriched so that added E. coli and indigenous populations were equivalent. Escherichia coli and total coliforms were enumerated using the Colilert‐18 Quanti‐Tray system. Patterns of E. coli survival and inactivation rates were virtually identical for indigenous strains in both mineral and organic sediments. The addition of E. coli strains from bovine, goose or deer faeces had relatively little impact on final E. coli concentrations, with the exception of deer‐borne E. coli populations in the organic sediment. Conclusion:  These results indicate that indigenous sediment‐borne E. coli strains are generally, or more, persistent than those deposited into sediments, including wildlife. Significance and Impact of the Study:  This is the first study on the survival of E. coli originating from wildlife faeces, in sediments, as opposed to bovine faeces or laboratory‐cultured strains. As wildlife are likely to be the primary source of E. coli in most non agricultural watersheds, an understanding of the persistence of these strains is important to understanding microbial water quality.     

An unusual clinical and pathological variant of malignant catarrhal fever in a white-tailed deer

A captive male white-tailed deer (Odocoileus virginianus) developed an acute illness over a 3-day period characterized predominantly by neurological, ocular and respiratory signs which were accompanied by prominent gross lesions of multiple organ systems. Histologically, a proliferative vasculitis consisting primarily of lymphocytic-lymphoblastic cellular infiltration was found in ocular, oral, respiratory, cardiac and neural tissues. The extensive nature of these infiltrations resulted in grossly apparent nodular foci in the lung, lymphoid tissue and myocardium which were suggestive of a lymphoproliferative disorder. This is contrasted to the more necrotizing nature of the vasculitis observed in other reported cases of malignant catarrhal fever in white-tailed deer. Although virus isolation was not attempted, serologic findings of antibodies to malignant catarrhal fever virus detected by indirect immunofluorescence and virus neutralization supported a diagnosis of malignant catarrhal fever in this deer.     

Antibodies to bluetongue and epizootic hemorrhagic disease viruses from white-tailed deer blood samples dried on paper strips.

The feasibility of using dried blood samples for serologic testing of white-tailed deer (Odocoileus virginianus) for antibodies to bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV) was tested with matched samples of serum and eluted dried whole blood. Results from matched serum virus neutralization (SN) tests indicated that a 1-ml elution from a 1- x 2-cm section of filter paper strip containing dried blood approximated a 1:10 serum dilution. Neutralizing antibody titers detected from 34 matched titrations of serum and dried blood samples were equivalent in 25 (74%) titrations and were within a single dilution in the remaining nine (26%) titrations. Eluted blood samples from SN-positive deer, however, did not produce detectable precipitin lines on agar gel immunodiffusion tests for antibodies to either BTV or EHDV. In a trial using serum and dried blood samples from 108 hunter-killed deer from five locations in Georgia (USA), antibody prevalence and serotype distribution results were similar. Use of dried blood samples for serologic testing for antibodies to BTV and EHDV provides a reliable alternative to serum but should be considered only when serum collection is not feasible.     

Assessment of in vitro fertility of deer spermatozoa by heterologous IVF with zona-free bovine oocytes

We have previously shown that the in vitro development of deer embryos differed according to the IVF conditions. The aim of the study was to use heterologous IVF with zona-free matured bovine oocytes to assess the in vitro fertility of 3 samples of deer semen (2 ejaculates from sika deer (Cervus nippon) and 1 pool of epididymal spermatozoa from red deer (Cervus elaphus)). The frozen/thawed semen samples were selected on Percoll gradient and resuspended in Tyrode modified medium supplemented with estrus sheep serum (0, 2, 20% v/v) or heparin (10 microg/mL). During 8 h of culture, the sperm motility index according to the post-insemination time (hpi) did not differ either between samples or between supplemented IVF media. In vitro matured zona-free bovine oocytes were inseminated in different IVF media with the semen samples. Penetration rates assessed at 15 hpi were optimal with 20% estrus sheep serum for sika deer ejaculates whereas 2% were sufficient to reach the maximum functionality of epididymal spermatozoa from red deer. The mean time of pronuclear formation was similar regardless of the semen sample. The precocity of the onset of the first S-phase in both pronuclei was characterized by Bromo-deoxy-Uridine exposures between 5 and 15 hpi in order to assess the developmental potential conferred by the semen sample (intrinsic value). As we previously observed in homologous IVF, this value seemed to be higher for the epididymal sperm sample.