Serological comparisons of antigenically related herpesviruses in cattle,red deer and goats

Serological comparisons were made using related herpesviruses from cattle (bovid herpesvirus 1), red deer (herpesvirus of cervidae 1) and goats (bovid herpesvirus 6) by virus neutralization and enzyme-linked immunosorbent assays. The test samples comprised field sera from British cattle, red deer and goats and sera from experimentally infected or immunized animals. Both the cervine and caprine viruses appeared to be more closely related to bovid herpesvirus 1 than they were to each other. Cattle sera reacted most strongly with the bovine virus and deer sera with the cervine virus. Antibodies to the caprine virus were not detected in the samples from British goats.     

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)     

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.     

Diarrheal condition in dogs associated with viruses antigenically related to feline herpesvirus

Viruses with properties consistent with herpesvirus were isolated from dogs with diarrhea. The viruses were shown to be antigenically related to feline herpesvirus-1 (FHV-1) by virus neutralization tests. It was also observed that a canine herpesvirus (CHV) prototype, D004, and two field isolates from fatal CHV infections in 2-week-old and 6-week-old puppies were neutralized at a low level by antiserum to FHV-1. Reciprocal neutralization tests with CHV antiserum against FHV-1 were negative. These results indicated that viruses related to FHV-1 can infect the dog and that there appears to be uni-directional virus neutralization of CHV by FHV-1 antibody.     

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.     

Construction and characterization of a glycoprotein E gene-deleted bovine herpesvirus type 1 recombinant

OBJECTIVE: To construct and characterize a recombinant glycoprotein (g)E gene-deleted bovine herpesvirus (BHV) type 1 (BHV-1). PROCEDURE: The BHV-1 gEgene-coding region and the flanking upstream and downstream sequences were cloned. The aforementioned cloned DNA was digested with suitable enzymes to release the amino terminal two thirds of that region, and was ligated to the beta-galactosidase (beta-gal) gene. The resulting plasmid DNA was cotransfected with DNA from full-length, wild-type (WT), BHV-1 Cooper strain of the virus. Recombinant viruses expressing beta-gal (blue plaques) were plaque purified and assayed further by blot hybridization for genetic characterization and by immunoblotting for reactivity against BHV-1 gE peptide-specific rabbit polyclonal antibody. One recombinant virus, gEdelta3.1IBR, was characterized in vitro and in vivo. The ability of the recombinant virus to induce BHV-1 neutralizing antibodies in infected calves was investigated by plaque-reduction tests. RESULTS AND CONCLUSIONS: The gEdelta3.1IBR virus contained a deletion in the viral gE gene-coding sequences where a stable chimeric reporter (beta-gal) gene was inserted. One-step growth kinetics and virus yield of the recombinant and parent viruses were similar, but early after infection, the recombinant virus yield was comparatively less. After intranasal inoculation, the recombinant gEdelta3.1IBR virus replicated in the upper respiratory tract of calves, but the amount of progeny viruses produced was hundredsfold reduced, and duration of virus shedding was shorter. Results of in vivo calf experiments and serum neutralization tests indicated that deleting the gE gene has little effect on inducing neutralizing antibodies against BHV-1, but is sufficient to reduce BHV-1 virulence in calves.     

Genetic and serologic analysis of feline cell-associated herpesvirus-induced infection of the urinary tract in conventionally reared cats

The genetic and antigenic nature of feline cell-associated herpesvirus (FeCAHV) was characterized by use of DNA restriction endonuclease analysis, and direct and indirect fluorescent antibody (FA) techniques. Serologic responses of 6 conventionally reared cats with induced FeCAHV urinary tract infection were retrospectively evaluated, using an indirect FA test. The EcoRI, HindIII, and Pst I restriction endonuclease cleavage patterns of FeCAHV DNA were similar to those of bovid herpesvirus 4 (BHV-4; DN599 strain) DNA. Specific fluorescence was observed when FeCAHV-inoculated cell monolayers were reacted with fluorescein-conjugated BHV-4 (DN599 strain) antiserum. Conversely, specific fluorescence was also observed when feline anti-FeCAHV serum and fluorescein-conjugated caprine anti-feline IgG was reacted with BHV-4 (DN599 strain)-infected cell monolayers. At postinoculation week 10, serum antibody titer in cats with FeCAHV-induced urinary tract infection ranged from 1:2,560 to 1:10,240, as measured by use of indirect FA testing. It was concluded that FeCAHV is a member of the BHV-4 group. In addition, the FeCAHV indirect FA test provides a sensitive and specific means of evaluating FeCAHV antibody concentration in exposed cats.     

Propagation and quantitation of animal herpesviruses in eight cell culture systems

A comparative study was carried out to determine the relative sensitivities of eight different cell culture systems to six different herpesviruses of animals. The cells used were: OFL (ovine fetal lung), ML (mink lung), FK (ferret kidney), PTK-2 (potoroo kidney), TEK (turkey embryo kidney), ED (equine dermal), BT (bovine turbinate), and PK15 (porcine kidney). The viruses tested were: PRV (pseudorabies) of swine, CPHV (caprine herpesvirus), IBRV (infectious bovine rhinotracheitis virus), DN-599 strain of bovine herpesvirus type 4, EHV-1 (equine herpesvirus), and CHV (canine herpesvirus). On the basis of virus titers obtained and the time of appearance of CPE (cytopathic effects), ML cells were found to be the most useful because of their sensitivity to all six viruses tested. BT and OFL cells were also found to be highly sensitive to all viruses with the exception of CHV.     

Crane hepatitis herpesviruses

Comparative studies were performed on crane herpesviruses obtained from two different enzootics in Austria and France. The examined viruses appear to be identical in their physico-chemical properties, morphology and biological reactions in ovo. The crane viruses are tentatively classified as beta-herpesviruses. Crane herpesvirus antisera produced in rabbits reacted in cross neutralization tests with each other and with a herpesvirus obtained from a bobwhite quail. No reactivity was observed with other avian herpesviruses and with human herpes simplex virus type 1 and 2.     

Immunological relationships between malignant catarrhal fever virus (alcelaphine herpesvirus 1) and bovine cytomegalovirus (bovine herpesvirus 3)

Strains of malignant catarrhal fever virus (alcelaphine herpesvirus 1 (AHV-1)) and bovine cytomegalovirus (bovine herpesvirus 3 (BHV-3)) were compared for serological relatedness by cross-titration in an indirect immunofluorescent (IIF) antibody assay. There was definite cross-reactivity between these 2 viruses, with heterologous sera staining intracellular and membrane antigens of infected cells. Heterologous antibody titres were approximately 50-fold lower than homologous titres and could be removed by absorption with either homologous or heterologous virus-infected cells, but not with uninfected cells. Regression analyses of IIF antibody titres to AHV-1 and BHV-3 virus in 3 groups of wild ungulate sera also indicated a serological relationship between these herpesviruses. In a cross-immunity trial, 2 of 3 cattle immunized with a BHV-3 virus and 2 of 3 cattle immunized with avirulent AHV-1 resisted challenge with virulent AHV-1-infected blood which killed 3 unimmunized controls. These results are discussed particularly with respect to the involvement of BHV-3 in malignant catarrhal fever.     

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.     

A bovine herpesvirus (BHV-4) as passenger virus in ethmoidal tumours in Indian cattle

A herpesvirus was isolated from tumours of the ethmoidal mucosa in two of three head of cattle in the State of Kerala, India. The virus designated M40 was cytopathic for a variety of cultured bovine and porcine cells and it did not kill suckling mice or chicken embryos. Sera from tumour-bearing cattle and goats reacted with the M40 virus. Immunofluorescence tests with FITC-conjugated IgG from a bovine monospecific antiserum to bovine herpesvirus 4 (BHV-4) stained the M40 virus specific antigen in infected cells. Experimental infection of goats with the M40 virus did not result in development of tumours. This virus is therefore considered to represent a "passenger" virus. A great similarity was found between restriction patterns of DNAs extracted from M40 virus and the strain 66-P-347, a reference strain of the BHV-4 group.     

Studies on bovine herpesviruses. Part 1. Isolation and characterization of viruses isolated from the genital tract of cattle

Herpesviruses, previously isolated from cattle (Theodoridis, 1978), were further studied and provisionally placed in the bovid herpesvirus 4 (BHV-4) group. Major differences were found between IBR-IPV (BHV-1) and BHV-4 virus strains. In MDBK cells, all BHV-4 strains started growing at the edges of the culture, the process progressing slowly until destruction of the cells was complete by the 10th day. BHV-4 strains failed to induce neutralizing antibodies in cattle, goats and rabbits. Only the addition of mineral oil adjuvant induced neutralizing and complement fixing antibodies in goats. BHV-1 strains, in contrast, produced very potent antisera in all these systems. Cross-neutralization tests indicated the existence of 2 distinct serological groups representing BHV-1 and BHV-4. The BHV-4 strains appear to be interrelated and they could not be grouped. A BHV-1 strain showed fixation of complement with the antisera of 6 BHV-4 strains. Electron micrographs showed an accumulation of nucleocapsids in the cytoplasm and an early release of virus particles due to cell destruction. Variation in incubation temperature had a significant effect on the particle formation. At lower temperatures, the number of enveloped particles in the cytoplasm increased. On the basis of the characteristics uncovered in this study, it is possible that all the BHV-4 strains represent one and the same virus which has undergone certain biological changes, thus illustrating a phenomenon which appears to be a characteristic of the herpesviruses.     

Biology of bovine herpesviruses

Cattle are the natural host of herpesviruses. Since now four different bovine viruses have been described as members of the family Herpesviridae. The prototype of the bovine herpesviruses, Bovine Herpesvirus type 1 (BHV-1), is the causative agent of infectious bovine rhinotracheitis (IBR), infectious pustular vulvovaginitis (IPV) and infectious balanoposthitis (IBP). The related BHV-5 is an exotic neurovirulent agent and like BHV-1 a member of the genus Varicellovirus, within the subfamily Alphaherpesvirinae. BHV-2, also an alphaherpesvirus but grouped into the genus Simplexvirus is the causative agent of bovine herpes mammilitis and pseudolumpy skin disease. In contrast, BHV-4, a member of the subfamily Gammaherpesvirinae, is not known to cause any disease. Beside bovine herpesviruses there are few other herpesviruses which can infect cattle. Infections of cattle with these herpesviruses have either clinical or diagnostic importance, based on a close antigenic relationship to BHV-1 of some ruminant herpesviruses. This article deals with the molecular virology of bovine herpesviruses and the pathogenesis of bovine herpesvirus infections and provides an overview over herpesviruses that can infect cattle.     

Antigenic Relationships of Equine Herpesvirus Strains Demonstrated by the Plaque Reduction and Neutralization Kinetics Test

The antigenic relationships among 50 strains of equine herpesvirus (EHV) were studied by neutralization tests using antisera prepared in rabbits against four EHV reference strains: types 2 and 3, cytomegalo-like virus 82-A, and our leukocyte isolant H-40. No distinctive antigenic differences among reference strains were demonstrated in reciprocal neutralization tests but each antiserum neutralized its homologous virus more rapidly than any heterologous strain. Forty-six EHV strains isolated from peripheral blood leukocytes of apparently healthy horses were antigenically indistinguishable from each other and from the four reference strains. Their high degree of antigenic relatedness suggests that these viruses are isolants of a single, widely distributed serotype of which type 2 (LK) strain is a typical representative.     

Comparison of the herpesviruses of cattle by DNA restriction endonuclease analysis and serologic analysis

Reference strains and field isolates of herpesviruses recovered from cattle in the United States were compared by restriction endonuclease (RE) analysis and the indirect fluorescent antibody test. As a result of these comparisons, 5 major biotypes of bovine herpesvirus (BHV) were defined. These types were (i) infectious bovine rhinotracheitis virus (BHV-1), (ii) bovine herpes mammillitis virus (BHV-2), (iii) malignant catarrhal fever (MCF) virus (herpesvirus alcelaphinae), (iv) the group of slow-growth isolates represented by the prototype strain Movar 33/63 (bovine cytomegalovirus candidate), and (v) the syncytia-forming Pennsylvania 47 strain. Bovine herpesvirus-1 and BHV-2 did not cross-react serologically with any other type of BHV tested. A low, but consistent level of serologic cross-reactivity was detected among MCF virus, the Movar group, and Pennsylvania 47. Several nonsyncytial, slow-growth strains, which were recovered from dissimilar clinical syndromes and were serologically related to Movar 33/63, exhibited similar DNA RE cleavage patterns, confirming their identity as members of a single type. There was no isolate from American domestic cattle similar to the African MCF virus, which has been sporadically isolated from exotic ruminants in the United States. The African MCF virus isolated during a MCF epizootic in a United States zoo exhibited some DNA RE cleavage differences in comparison with the MCF virus world prototype strain WC 11, indicating that strain diversity exists within this biotype.     

Aujeszky's disease ELISA: cross-reactions with other herpesvirus antisera

The production of antibodies in pigs to 11 herpesviruses was investigated in relation to their ability to cross-react with Aujeszky's disease virus (suid herpesvirus 1--SHV1). Of the herpesviruses tested only two, sheep herpesvirus (caprine herpesvirus 1) and dog herpesvirus (canid herpesvirus 1), failed to produce homologous virus antibodies. Only the antibodies to bovine herpesvirus 1 (BHV1) produced a cross-reaction by SHV1 enzyme-linked immunosorbent assay (ELISA). No SHV1 neutralizing antibodies were detected in any of the herpesvirus antisera. A cross-reaction with SHV1 by a serum from a pig naturally infected with BHV1 or with any of the other herpesviruses tested was considered unlikely.     

Antigenic relatedness of pigeon herpes encephalomyelitis virus to other avian herpesviruses

Pigeon herpes encephalomyelitis virus (PHEV) was compared with seven avian herpesviruses for antigenic relatedness using monospecific antisera and the indirect fluorescent-antibody (IFA), agar-gel-immunodiffusion, and serum-neutralization tests. No antigenic relationship was detected between PHEV and Marek's disease virus, turkey herpesvirus, infectious laryngotracheitis virus, and duck enteritis virus. A common precipitating antigen was detected between the PHEV and pigeon herpesvirus (PHV), owl herpesvirus (OHV), and falcon herpesvirus (FHV). These four viruses also cross-reacted in the IFA test. Weak neutralizing activity was detected only between PHV antiserum and PHEV. These results suggest that the PHEV should be classified as a herpesvirus related to, but distinct from, the PHV-OHV-FHV group of viruses with which it shares common antigens.     

Seroprevalence of OHV-2, BVDV, BHV-1, and BRSV in ranch-raised bison (Bison bison).

Serum samples were collected at slaughter from 226 24-30-month-old American bison (Bison bison) bulls from Kansas, Minnesota, North Dakota, and Manitoba and assayed for antibodies to ovine herpesvirus type-2 (OHV-2), bovine viral diarrhea virus (BVDV), bovine herpesvirus type-1 (BHV-1), and bovine respiratory syncytial virus (BRSV). Antibodies were detected by serum neutralization for BVDV, BHV-1, and BRSV, while antibodies to OHV-2 were detected by competitive inhibition-ELISA (CI-ELISA). Detectable antibodies were found against all viruses: 10 of 226 (4.40%) against OHV-2, 125 of 226 (55.3%) against BVDV, 99 of 226 (43.8%) against BHV-1, and 208 of 226 (92.0%) against BRSV. Titers from 93.6% of the BVDV-positive animals, 79.8% of the BHV-1-positive animals, and 98.1% of the BRSV-positive animals were > or = 1.25. These data indicate that a low percentage of clinically normal bison are seropositive for OHV-2 while a high percentage of bison sampled are seropositive for BVDV, BHV-1, and BRSV.     

A study of a herpesvirus isolated from dairy cattle with a history of reproductive disorders

Three strains of herpesvirus were recovered from cows with vulvovaginitis. The three isolates (85/BH 16TV, 85/BH 17TV, 85/BH 18TV), when compared by cross serum neutralization (SN) tests, were found to be antigenically identical. They were serologically distinct from infectious bovine rhinotracheitis (IBR) virus and Bovid herpesvirus 2 (BHV2), while they cross reacted with bovine herpesvirus DN-599. Besides the serologic aspects, the three isolates appeared to share common biological, physical and morphological properties with the newly recognized bovine herpesviruses, of which DN-599 is a representative strain.