A serological comparison of some animal herpesviruses

Bovine herpesvirus 1 (BHV-1) isolates (Cooper-type strain 4975 and Oxford) were compared in neutralization tests with the bovine herpesvirus 4 (BHV-4) isolate (85/16 TV) and the herpesviruses of red deer (D2839/1) and goats (E/CH). Hyperimmune antiserum was prepared in rabbits against the plaque-selected viruses and endpoint and kinetic neutralization test were made. BHV-4 was clearly different from the other four viruses. The closely-related BHV-1 strains were also related in these tests to the red deer herpesvirus. The Oxford strain seemed rather closer antigenically than the Cooper-type strain to the red deer herpesvirus. Antiserum to the caprine herpesvirus failed to neutralize either BHV-1 strain or red deer virus, but antiserum to the Cooper-type and red deer herpesviruses did neutralize caprine virus to a limited extent.     

Isolation and characterisation of cervine herpesvirus-1 from red deer semen

AIM: This communication describes the isolation of herpesvirus during routine export examination of semen collected from red deer stags in New Zealand. METHODS: Virus isolation was carried out using bovine embryonic lung (BEL) cells and viruses were characterised by direct immunofluorescense, restriction-fragment-length polymorphism analysis (RFLP), polymerase chain reaction (PCR) analysis and nucleotide sequencing. RESULTS: Herpesvirus was isolated from red deer semen on 2 different occasions from different animals. In both cases the virus was identified as cervine herpesvirus-1 (CvHV-1), based on RFLP, PCR and sequence analysis. Nucleotide sequence analysis of the glycoprotein-D gene showed 99.7% homology to the Banffshire strain of CvHV-1 and 89.5%, 89.2%, 85.3% and 79.6% homology to bovine herpesvirus 1.2 (BoHV-1.2), bovine herpesvirus 1.1 (BoHV-1.1), cervine herpesvirus-2 (CvHV-2) and caprine herpesvirus-1 (CpHV-1), respectively. CONCLUSION: This is the first time that CvHV-1 has been isolated in New Zealand. Its inclusion in serological surveys will allow the prevalence of CvHV-1 in the red deer population to be assessed in this country. The clinical significance of CvHV1 infection in New Zealand red deer herds has yet to be determined.     

Antibody to alcelaphine herpesvirus-1 (AHV-1) in hamsters experimentally infected with AHV-1 and the 'sheep-associated' agent of malignant catarrhal fever

Malignant catarrhal fever was induced in four groups of hamsters by the inoculation of cells infected with either the C/500 isolate of alcelaphine herpes-virus-1 (AHV-1) or the sheep-associated agent derived from cattle, red deer or Père David's deer. Using an indirect immunofluorescence assay, antibody to AHV-1 was detected in sera of clinically affected animals of all four groups. The reaction of sera from hamsters affected with malignant catarrhal fever induced by AHV-1 caused diffuse cytoplasmic staining while that from sera of hamsters with the sheep-associated form of the disease stained particulate nuclear antigens. Tests employing three other bovid herpesviruses were negative and no reaction was found with sera from normal hamsters. These studies provide convincing evidence that a virus antigenically related to AHV-1 is the cause of sheep-associated malignant catarrhal fever and that the same virus probably causes this form of the disease in both cattle and deer.     

Serologic and virologic investigations into pestivirus infection in wild and domestic ruminants in the Pyrenees (NE Spain)

An outbreak of disease associated to a border disease virus was described in the Southern chamois (Rupicapra pyrenaica) in Spain in 2002. Sera and/or spleen samples from 57 mouflon, 15 red deer, 21 roe deer, 3 fallow deer, 55 sheep, 32 cattle, and 68 goats sharing the chamois habitat were studied. An antibody ELISA test yielded an inconclusive result in 2 mouflon and positive results in 5 goat sera. Comparative virus neutralization tests were performed on the 2 inconclusive mouflons, 3 of the 5 seropositive goats, 55 sheep and 32 cattle, using 6 pestivirus strains. Positive results were obtained in 1 mouflon, 2 goats, 69% of sheep and 78% of cattle. Virological investigations performed with an antigen ELISA test yielded negative results in 21 goats and 39 mouflons, the result in 1 mouflon being inconclusive. PCR performed on 12 goats and the inconclusive mouflon gave negative results. These results suggested that it is unlikely that chamois BDV is infecting wild and domestic ruminants.     

Latent herpesvirus infection in red deer: characterization of a specific deer herpesvirus including comparison of genomic restriction fragment patterns

Glucocorticoid treatment of imported red deer (Cervus elaphus), seropositive to Infectious Bovine Rhinotracheitis (IBR) virus, reactivated a latent herpesvirus infection, which was transmitted to a seronegative deer with a fatal outcome. However the virus did not spread to cattle housed in close contact with the infected deer, and serological indication og infection in the cattle was observed only on direct nasal installation of virus. The virus isolate had characteristics in common with other Alpha herpesviruses and especially the Bovid Herpesvirus type 1 (BHV-1) but distinguished itself from the latter by its host specificity, serological reaction and genomic restriction fragment pattern (RFP). The host specific red deer herpesvirus was tentatively designated Cervid Herpesvirus type 1 (CHV-1). It was concluded that CHV-1 seropositive deer can be a threat to red deer farming, while in cattle the infection may only cause minor inconvenience through interference with the serological IBR diagnosis.     

Caprine herpesvirus-2-associated malignant catarrhal fever in white-tailed deer (Odocoileus virginianus).

A subacute disease presenting primarily as alopecia and weight loss occurred in 2 white-tailed deer (Odocoileus virginianus) on farms in Minnesota and in Texas. A presumptive diagnosis of malignant catarrhal fever (MCF) was made on the basis of histological lesions. Antibody against an epitope conserved among the MCF group viruses was detected in the serum of both deer. DNA samples from the deer were subjected to a variety of PCR amplifications. Alignment of the amplified sequences from the diseased animals revealed that they were 100% identical to each other and to the same DNA fragment from the newly recognized member of the MCF virus group endemic in domestic goats (Capra hircus), provisionally named caprine herpesvirus 2 (CpHV-2). A seroprevalence survey from one of the deer farms showed a high rate of subclincal infection in the deer population. This study provides further confirmation that CpHV-2 is a pathogen, at least for deer, and emphasizes the risk of loss from MCF when mixing cervids with goats.     

Infection of sheep and goats with bovid herpesvirus 2

Sheep and goats were shown to be susceptible to experimental infection with bovid herpesvirus 2 (BHV2), administered by either the intradermal or intravenous route. Lesions developing in sheep following intradermal inoculation of virus were similar to those in cattle inoculated intradermally, whereas the lesions in goats resolved without ulceration or scabbing. Disseminated circumscribed skin lesions developed in sheep and goats given BHV2 by the intravenous route. These lesions resolved in four to eight days without significant effect on the skin. BHV2 was isolated from skin lesions of sheep, goats and cattle that were infected intravenously, from sheep and cattle infected intradermally and from the leucocytes of the three species following intravenous inoculation of virus. Latent infection of sheep and goats was demonstrated following corticosteroid treatment 60 days after infection.     

Vulvovaginitis of goats due to a herpesvirus

Two concurrent outbreaks of genital disease in goats were associated with infection by a herpesvirus that was isolated from vulval and vaginal lesions of affected does. Serum neutralising antibody to the virus was present both in goats with the clinical disease and some unaffected goats. Of 19 goat herds examined only 4 had serum neutralising antibody positive goats with low (5%) to high (60%) incidence of infection. The virus isolate was characterised as a herpesvirus on its physico-chemical and morphological features. It contained DNA and was inactivated at low pH and by treatment with lipid solvents and trypsin. The virus particles were icosahedral, consisting of a nucleocapsid surrounded by an envelope membrane and measured approximately 150 nm in diameter. The virus was serologically related to a New Zealand isolate of caprine herpesvirus (NZ-CpHV), associated with similar genital disease, and was distinct from bovine herpes virus-1 (BHV-1) showing a one way neutralisation pattern.     

Pathogenicity of a caprine herpesvirus.

A herpesvirus isolated from neonatal Angora kids (Capra hircus) with a relatively severe generalized infection was shown to be infective for adult goats as well as for kids. However, the virus lacked pathogenicity for either lambs or calves. Although a nonreciprocating serologic overlap exists between the caprine virus and the antibody to bovine herpesvirus (BHV-1), results of cross immunity tests in calves indicated that the 2 viruses are immunologically distinct. On the basis of these findings, the caprine virus seems to be a specific pathogen of goats. Accordingly, the designation of Herpesvirus caprae or caprine herpesvirus 1 seems taxonomically appropriate.     

Characterization of bovine herpesviruses isolated from six sheep and four goats by restriction endonuclease analysis and radioimmunoprecipitation

Viral DNA from 10 herpesviruses isolated from 6 sheep and 4 goats were examined by restriction endonuclease analysis with respect to their relatedness to one another; to bovine herpesvirus type 6 (BHV-6), also known as caprine herpesvirus; and to 2 strains of bovine herpesvirus type 1 (BHV-1), known as infectious bovine rhinotracheitis virus (IBRV) and infectious pustular vulvovaginitis virus (IPVV). Viral proteins from the isolates were examined by radioimmunoprecipitation with anti-BHV-1/IBRV gnotobiotic calf (bovine) serum, anti-BHV-1/IBRV bovine hyperimmune serum, and anti-BHV-6 rabbit serum to evaluate their antigenic relatedness to each other. The goat isolates were obtained from animals with various disease conditions including respiratory tract disorders, vulvovaginitis, and wart-like lesions on the eyelid. The other isolates were from domestic sheep and came from aborted fetuses or from sheep with fatal pneumonia or proliferative lesions around lips and nose. All of the goats and 4 of the sheep from which the viral isolates were obtained had comingled with cattle. Purified DNA from each of the 10 field isolates and from BHV-1/IBRV, BHV-1/IPVV, and BHV-6 caprine herpesvirus was cleaved with restriction endonuclease Pst I. Five of 6 sheep isolates and 3 of 4 goat isolates yielded unique restriction patterns, ie, patterns that differed from each other by one or more bands. Sheep isolate DNA patterns were different from goat isolate patterns, and all restriction endonuclease analysis patterns were similar to the pattern for BHV-1/IBRV, but different from that for BHV-1/IPVV or for BHV-6.(ABSTRACT TRUNCATED AT 250 WORDS)     

Production and characterization of monoclonal antibodies to cervine herpesvirus-1.

Fourteen hybridoma cell lines secreting monoclonal antibodies (Mab) to cervine herpesvirus-1 (CerHV-1) produced following the fusion of NSO myeloma cells with splenocytes of BALB/c mice previously immunized with gradient purified CerHV-1 were selected using an indirect enzyme-linked immunosorbent assay (ELISA) employing CerHV-1 antigen and tested by the ELISA against four other ruminant alphaherpesviruses from cattle (bovine herpesvirus type 1.1 and 1.2) goat (caprine herpesvirus-2) and reindeer (rangiferine herpesvirus-1). Comparison of all five ruminant alphaherpesviruses with these Mabs confirmed their close antigenic relationships, with two Mabs reacting against all viruses. Ten Mabs which were able to differentiate between the viruses reacted with a 64 kDa polypeptide in a western blot. Four Mabs including two specific only for CerHV-1 with neutralizing activity against the virus used for immunization were directed against a 74 kDa viral protein.     

Bovid herpesvirus 2 latency: failure to recover virus from central sensory nerve ganglia.

Latent bovid herpesvirus 2 was sought in sensory ganglia, epithelium and lymph nodes from cattle having antibodies against bovid herpes virus 2. Tissues from eight animals were maintained in vitro as explants for 49-72 days during which all expended media was tested for virus. Three animals were pretreated with corticosteroids prior to slaughter. Infectious bovine rhinotracheitis virus was recovered from one animal, but bovid herpesvirus 2 was not detected.     

The virulence of British isolates of bovid herpesvirus 1 in relationship to viral genotype.

Six strains of bovid herpesvirus 1 isolated from British cases of infectious bovine rhinotracheitis (IBR) were inoculated intranasally into calves. The clinical, virological and serological responses were measured and comparisons made between virus strains. Calves infected with viruses of subtype 1 developed more severe clinical signs and excreted higher titres of virus than calves infected with subtype 2b strains. The findings could be related to the history of IBR in Great Britain.     

Dictyocaulus species: cross infection between cattle and red deer

Aim: To discover whether cross infection between red deer (Cervus elaphus) and cattle is possible with either a bovine isolate of the cattle lungworm, Dictyocaulus viviparus, or with a cervine isolate of the lungworm, Dictyocaulus eckerti which is thought to be maintained primarily in deer. Method: Twelve cattle and 12 red deer were reared parasite-free from birth. At 3-4 months of age, half of each species (n=6) were experimentally infected with D. viviparus and the other half with D. eckerti. The course of infection was monitored for 34 days, after which the animals were slaughtered and the lungs removed to assess levels of infection. Results: Faecal larval counts demonstrated that patent Dictyocaulus infections occurred in all groups. At necropsy, adult worms were found in the lungs in all groups except the cattle that were infected with D. eckerti. The largest numbers of adult worms were found in the red deer infected with D. eckerti. Conclusion: It was demonstrated that both cattle and red deer could be infected with either D. viviparus or D. eckerti. However, D. eckerti larvae that originated from deer established more successfully in deer and D. viviparus larvae that originated from cattle established more successfully in cattle.     

Caprine herpesvirus-2 in association with naturally occurring malignant catarrhal fever in captive sika deer (Cervus nippon).

Three female sika deer from a single captive herd were submitted for postmortem examination over a 139-day period. The first 2 deer submitted were reported to have lost body mass for 20 days to 1 month before euthanasia. One of these deer had diarrhea, the other had a crusting dermatitis on the nasal planum and inner aspects of both pinnae. The third hind did not have any signs of disease before it was found seizuring and was immediately euthanatized. Microscopically, all 3 animals had a lymphocytic vasculitis typical of malignant catarrhal fever (MCF), with the most severe lesions in the brain. All 3 deer were polymerase chain reaction (PCR) positive for caprine herpesvirus 2 (CpHV-2) and were negative for ovine herpesvirus 2 (OHV-2). Two healthy goats that were housed adjacent to the deer were also PCR positive for CpHV-2 and PCR negative for OHV-2. The CpHV-2, PCR amplicons from the hinds, and the 2 healthy goats had an identical single base polymorphism. A male sika deer that was housed with the hinds and a fawn from 1 of the hinds remained asymptomatic and were PCR negative for CpHV-2. This represents the first report of mortality with MCF-like lesions in association with CpHV-2.     

Experimental respiratory infection of goats with caprine herpesvirus and Pasteurella haemolytica

Groups of six male goats were inoculated intratracheally and intranasally with either caprine herpesvirus followed 6 days later by Pasteurella haemolytica, canine herpesvirus alone or P. haemolytica alone. Pneumonic lesions were observed in five of the six goats inoculated with caprine herpesvirus followed by P. haemolytica and in three of the six goats inoculated with P. haemolytica alone, but were not observed in goats inoculated with caprine herpesvirus alone or in non-infected controls. Pasteurella haemolytica was isolated from seven of eight lungs with pneumonia, but only from one of sixteen lungs without pneumonia. The lesions ranged from fatal acute exudative necrotising pneumonia to predominantly proliferative pneumonia. Half of the caprine herpesvirus-inoculated goats developed a clinical catarrhal rhinitis five days post-inoculation and the only virus-specific histopathological lesion was a mild tracheitis. Canine herpesvirus was recovered from the nasal swabs of all caprine herpesvirus- inoculated goats developed a clinical catarrhal rhinitis five days post-inoculation and the only virus-specific histopathological lesion was a mild tracheitis. Canine herpesvirus was recovered from the nasal swabs of all canine herpesvirus-inoculated goats and from the lungs of three goats inoculated with caprine herpesvirus alone. The experimental inoculations demonstrated that P. haemolytica alone can produce pneumonia in goats. In addition, the study showed that caprine herpesvirus readily proliferates in the upper respiratory tract and lungs of goats but the role of caprine herpesvirus in the aetiology of pneumonia remains uncertain.     

The gp135 of caprine arthritis encephalitis virus affords greater sensitivity than the p28 in immunodiffusion serology

The 135,000 mw glycoprotein (gp135) and the 28,000 mw internal protein (p28) of caprine arthritis encephalitis virus are major viral constituents in precipitin lines formed between crude antigen preparations and sera from infected goats. In testing 307 goat and sheep sera, 118 samples were positive in a gp135 assay and only 82 were positive in a p28 assay. However, some goat sera were found which reacted only with the p28 and therefore testing for antibody against both proteins may be necessary to identify a maximum number of virus infected goats by immunodiffusion.     

Isolation of caprine herpesvirus type 1 in Spain

Two strains of caprine herpesvirus type 1 (CpHV-1) were isolated after the experimental reactivation of two seropositive goats in Spain. Viral DNA from these isolates was compared with DNA from bovine herpesvirus type 1 and CpHV-1 reference strains by restriction endonuclease analysis. The two Spanish isolates were closely related but could easily be distinguished from each other and from the reference strains.     

Seroprevalence of antibodies against bluetongue virus in animals imported to Poland from EU countries

The aim of this study was to determine the seroprevalence of BTV-specific antibodies in animals imported to Poland from EU countries after 15 June 2006. From 1 January 2007 to 22 January 2008, a total of 10719 samples of sera collected from cattle, goats and fallow deer were tested. Sera were screened using the highly sensitive and specific c-ELISA test and positive results were confirmed by the AGID assay. Out of 10719 sera, 30 (0.28% of the total number of samples) were found to be positive in both tests applied. All of 21 seropositive cattle specimens were imported to Poland from Germany whereas 9 seropositive fallow deer were of Dutch origin. In conclusion, it can be stated that because BTV situation in Europe is getting worse, implemented surveillance studies should be continued to monitor the actual BT status in Poland.     

An outbreak of vulvovaginitis in goats caused by a caprine herpesvirus

A caprine herpesvirus related to infectious bovine rhinotracheitis virus but immunologically distinct from that virus was isolated from an outbreak of vulvovaginitis in a herd of Saanen goats. The morbidity rate was 52.5%, with 21 of 40 does showing clinical signs. The lesions healed rapidly with only two goats showing lesions two weeks after the disease was first detected. No effect on subsequent reproductive performance was observed. The mode of transmission of the virus was believed to be venereal.