Herpes-type virus particles associated with a fungus.

A cultutre of the fungus Thraustochytrium, isolated from an estuary, was infected by ani enveloped virus. The nucleocapsid measured 110 nanometers in diameter and containied a core of DNA. The virus replicated in the nucleus. These findings stronigly suggest that the particles are a herpes-type virus.

Virus-free adenocarcinoma of the frog (summer phase tumor) transcribes Lucké tumor herpesvirus-specific RNA.

[(3)H]RNA isolated from "virus-free," summer phase, renal adenocarcinomas of Rana pipiens labeled with [(3)H]uridine hybridizes with DNA of herpesvirus particles isolated from the winter phase Lucké tumor. Transcription of the herpesvirus genome in virus-free tumors provides additional evidence for the viral etiology of this tumor.

Morphogenesis of marek's disease virus in feather follicle epithelium.

Three evaluative systems, immunodiffusion, fluorescent antibody (FA), and electron microscopy (EM), were used to follow the morphogenesis of Marek's disease virus in inoculated chickens. Of the three, EM and FA were the most sensitive in detecting early stages of infection. Virus particles were found in skin biopsy specimens as early as 12 days post inoculation. Immature naked particles appeared first in the nucleus; later particles were enveloped in the cytoplasm and enclosed in cytoplasmic inclusion bodies. No evidence for continued virus replication was seen in feather follicles after an initial burst of heavy virus production, which lasted several weeks. Residual virus, however, was found occasionally in cytoplasmic inclusion bodies within keratinized material near the feathers. This was believed to contribute to the long-term shedding of infectious virus into the environment.

Virion and nonstructural polypeptides of human herpesvirus-6.

Human herpesvirus-6 (HHV-6) was purified from HHV-6-infected mononuclear cells from cord blood or infected culture medium. Virion was found to have at least 29 polypeptides ranging from 280 to 30 kDa. Six polypeptides were identified in the envelope fraction. Twenty-five viral polypeptides were also identified in HHV-6 infected cells by immunoprecipitation with immune serum.

Use of lectins to detect and differentiate subtypes of Marek's disease virus and turkey herpesvirus glycoproteins in tissue culture.

Biotinylated lectins in conjunction with an avidin-biotin-peroxidase complex were used for the first time to reveal glycoproteins in chicken and duck embryo fibroblasts infected with three prototype members of the avian herpesvirus group, Marek's disease virus serotypes 1 and 2 and turkey herpesvirus. By using a panel of 10 lectins, a pattern of reactivity emerged which was both group- and type-specific. Morphological details of the lectin-stained cells include cytoplasmic granulation, capping and bleb-like protrusions of the cell membrane. Although no antibody is necessary for the reaction, this novel approach allows specific detection of the glycan moieties of viral glycoproteins as they are synthesized during infection.

Further biological, serological and biochemical characterization of North American, European and Southeast Asian strains of bovine herpesvirus 1 compared with other alphaherpesvirinae members.

Hemagglutination activity, structural protein profiles and neutralization assays were used in a comparative study of bovine herpesvirus 1 strains from the U.S.A., Canada, Great Britain, Denmark and Malaysia with equine, feline and human herpesviruses in order to further characterize the bovine herpesvirus 1 hemagglutinin. Bovine herpesvirus 1 strains of different geographical origins all showed hemagglutinating activity for mouse erythrocytes; furthermore, feline herpesvirus 1 was also shown to hemagglutinate mouse erythrocytes. Analyses of partly purified viruses showed that a distinctive and specific polypeptides profile is associated with each species of herpesviruses used in our study; strains of bovine herpesvirus 1 from North America, Europe and Southeast Asia however, presented a remarkable similarity as to their electrophoretic protein patterns. A protein similar to the 97-kDa bovine viral hemagglutinin was not identified with the hemagglutinating feline herpesvirus. An important neutralization epitope on the bovine viral hemagglutinin was also not found on feline, equine and human herpesviruses but was identified on all bovine strains tested from North America, Europe and Southeast Asia stressing the importance of the bovine hemagglutinin for eventual prophylactic purposes.

Restriction endonuclease patterns of some European and American isolates of avian infectious laryngotracheitis virus.

Eleven isolates of infectious laryngotracheitis virus (nine European and two American) were compared by restriction endonuclease analysis of their DNA after radiolabeling with 32P. Digestion with KpnI gave identical cleavage patterns for all the European isolates, but the two American viruses (one field and one vaccine) showed some differences from them and from each other. In the case of the American vaccine strain, however, these differences were only minor. After BamHI digestion, only the American field isolate appeared to be different, whereas with HindIII, all 11 isolates were identical.

Viral encephalomyelitis of pigeons. VI. Some physico-chemical properties of the virus and extracted viral DNA.

Pigeon herpes encephalomyelitis virus (PHEV) was stable at -70 C for at least 4 months. When stored at -20 C, the virus lost 80% of its infective titers in 4 months. When stored at -10 C, however, the titers decreased rapidly; no detectable virus remained within 12 weeks. PHEV was thermolabile: it was completely inactivated at 56 and 60 C for 10 and 2 min respectively. It was also killed by 1% cresol and 2% sodium hydroxide for two hr and 2% septol for 24 hr. Two-percent phenol or formaline for 2 hr, however, significantly decreased virus infective titers. Phenol-purified DNA extracted from PHEV showed an ultraviolet spectrum of typical nucleic acids that had ratios of absorbancies at 265 nm/280 nm between 2 and 2.3. The extracted viral DNA was infectious in chorioallantoic membrane and chick embryo fibroblast cell cultures, but it was not noninfectious when given to pigeons. DNA infectivity was destroyed by DNAse but not RNAse treatment. Extracted DNA was not neutralized by antiserum against the intact virus, and it lost its infectivity property when heated at 70 C for 10 min.

Isolation of bovine herpesvirus-3 from African buffaloes (Syncerus caffer).

Eleven virus isolations were made from the blood of 45 free living healthy African buffaloes by long term cocultivation of their leucocytes with bovine thymus or spleen cells. The isolates were indistinguishable from each other or from herpesviruses isolated from a severely ill buffalo calf and from a dead buffalo. These viruses possessed the characteristics of the bovine herpesvirus-3 (BHV-3) group and were indistinguishable by serology and restriction endonuclease analysis from the BHV-3 type strains Movar 33/63 and DN599. There was a 93.6 per cent prevalence of indirect immunofluorescent antibody to BHV-3 in the sera of 94 buffaloes in the sample population. No clinical signs or viraemia were detected in five cattle inoculated with 10(8.7) log10 TCID50 of the isolate from the sick buffalo calf. Two of three cattle hyperimmunised with this virus resisted challenge with malignant catarrhal fever herpesvirus, which proved fatal for the other immunised animal and for three control cattle.

[Analysis of B-lymphotropic oncogenic herpesvirus of Macaca arctoides]. / Analiz B-limfotropnogo onkogennogo virusa gerpesa makakov burykh.

Some physico-chemical and molecular-biological parameters of B-lymphotropic herpes virus of Macaca arctoides (HVMA) have been analyzed. The buoyant density of virions and nucleocapsids in a gradient of the percoll density is 1.10 and 1.14 g/ml, respectively, while that of DNA in CsCl gradient is 1.717 g/ml, which is typical for herpes viruses of primates. At the same time the homology of DNA HVMA with DNA EBV and DNA HVP is 30-45%. Differences in restrictions sites between DNA HVMA, DNA HVP and DNA EBV are determined. The intraspecific heterogeneity of HVMA strains has been detected according to restriction sites of DNA. The HVMA genome size is about 170 kb. The DNA HVMA is colinear to the EBV genome, which is also typical of B-lymphotropic herpes viruses of primates.

Application of cloned fragments of equine herpesvirus type-1 DNA for detection of virus-specific DNA in equine tissues.

Tissue specimens obtained from equine herpesvirus-1 (EHV-1), subtype 1-infected aborted foetuses were analysed for the presence of virus DNA by means of Southern blot and dot blot hybridisations. The specificity of the methods was confirmed although the sensitivity was inferior to classical techniques such as virus isolation. However, the possibility of detecting the state of the virus DNA and the ability to distinguish between subtypes were important features, and the dot blot method was shown to have potential for a rapid diagnostic test. This report demonstrates some potential practical applications of hybridisation methods for studying the pathogenesis and epidemiology of EHV-1 but also reveals limitations of the techniques.

Channel catfish virus: use of nucleic acids in studying viral relationships.

Restriction digestion patterns were used to determine differences in the DNA of various isolates of channel catfish virus (CCV). All viruses were different from each other and from the type strain of CCV. The differences in the digestion patterns were used to relate the viruses quantitatively as to sequence divergence between all pairs of viruses. A range of values from 104 nucleotide changes to 1,690 nucleotide changes/total DNA of CCV was found for the various pairs. A cladistic analysis produced a phylogenetic network relating the viruses by possible ancestory. This network indicated that some viruses were relatively more separated from other viruses that were clustered in the network. A phenetic analysis indicated that the viruses that were clustered in the network were also reasonably similar to one another.

Serologic and molecular comparisons of several equine herpesvirus type 1 strains.

The molecular and serologic relatedness of 2 recent respiratory tract isolates of equine herpesvirus type 1, designated T1 and T2, were compared with the Army 183, Kentucky-A hamster-adapted (KyA-ha), and L-M cell-adapted (KyA-LM) strains. Electrophoresis in polyacrylamide gels revealed differences in virion structural proteins among 4 purified strains. Seven envelope glycoproteins (molecular weight of 93,000, 65,000, 62,000, 60,000, 36,000, 20,000, and 18,000) corresponding to virion proteins 13, 16, 17, 18, 23, 25, and 26a, respectively, found in both the Army 183 and KyA-ha strains had slightly different molecular weight counterparts in both the T1 and T2 isolates, which had identical structural protein profiles. virion protein 19 (58,000 daltons), a nonglycosylated protein, was present in reduced amounts in the respiratory tract isolates, whereas virion protein 8a (200,000 daltons) was absent. Virion protein 8a, an envelope glycoprotein, was only present in the KyA-ha strain. The T1 and T2 isolates were not neutralized by equine herpesvirus type 2 antiserum and revealed little cross-neutralizatio with the Army 183 and KyA-ha strains in plaque-reduction neutralization tests. Restriction endonuclease cleavage maps of viral DNA revealed a similar, but not identical, number and size of DNA fragments between T1 and T2 isolates. Likewise, DNa profiles of Army 183, KyA-ha, and KyA-LM were also similar to each other, but vastly different from the respiratory tract isolates.

Vertical transmission of channel catfish virus.

Channel catfish broodfish were examined nondestructively for the presence of channel catfish virus (CCV), using a nucleic acid-probing technique. A dot blot assay was tested and shown to be accurate and rapid in the diagnosis of CCV. Two CCV-positive fish were successfully mated, and fertilized eggs were collected. Offspring that hatched were tested and were shown to have CCV. This result indicated vertical transmission of CCV, a herpesvirus.

Presence of Herpesvirus ovis DNA sequences in cellular DNA from sheep lungs affected with jaagsiekte (pulmonary adenomatosis).

To investigate further the possible involvement of Herpesvirus ovis in the aetiology of jaagsiekte, the kinetics of reassociation of viral DNA and DNA isolated from tumour tissue as well as from cell cultures derived from it were studied. Although DNA-DNA hybridization could be demonstrated in 2 cases of jaagsiekte, no correlation was found between the presence of Herpesvirus ovis genome sequences and the occurrence of the disease.

Heterogeneity of pseudorabies virus studied by isoelectric separation.

Heterogeneity of four lines derived from a single strain of pseudorabies virus (PRV) was studied by isoelectric separation. The individual lines differed in the distribution pattern of infectious fractions separated in the pH range from 3.5 to 10.0. All virus fractions of the virulent and attenuated lines were infectious, but some fractions of the highly attenuated avirulent line were devoid of infectivity. Infectivity of the fractions isolated from the individual PVR lines depended on the degree of attenuation; the higher degree of attenuation, the greater the differences in infectivity of the isolated fractions. The possible causes of the different distribution of infectious fractions obtained by isoelectric separation from virus lines differing in biological properties are discussed. The relationship between physico-chemical and biological properties of PRV is stressed along with the possibilities of applying isoelectric separation in narrow pH gradients.

Comparative polypeptide analysis of human, murine and strigis herpesviruses with murine cytomegalovirus by polyacrylamide gel electrophoresis.

The polypeptide composition of five purified murine herpesvirus (MHV) strains grown in a stable line of rabbit embryo fibroblasts (REF) was analysed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and compared with herpes simplex virus type 1 (HSV-1). About 24 structural polypeptides of molecular mass ranging from 275,000 to 25,000 were identified in MHV and HSV-1. The polypeptide profiles of MHV and HSV-1, showed a close similarity. The polypeptides of MHV were further compared with those of HSV-1, HSV-2, herpes virus strigis (HVS) and murine cytomegalovirus (MCMV). Differences were found between herpesviruses of different origin and MCMV. SDS-PAGE analysis of the six strains of MCMV labelled with 14C-amino acid hydrolysate also revealed differences in electrophor eticprofiles of MHV and MCMV proteins, what was confirmed by densitometric scanning of HSV-1, MHV and MCMV.

Molecular epidemiology and pathogenesis of some equine herpesvirus type 1 (equine abortion virus) and type 4 (equine rhinopneumonitis virus) isolates.

Representative strains of EHV isolated from an aborted foetus and from a horse with rhinopneumonitis in New Zealand had restriction endonuclease DNA fingerprints typical of those usually associated with these syndromes elsewhere and now designated EHV1 and 4 respectively. EHV1 was isolated from the brain and spinal cord of a 4-year-old gelding that died of myeloencephalitis. A mare on the same farm, at about the same time as the gelding developed myeloencephalitis, aborted and EHV1 was isolated from the tissues of the aborted foetus. Restriction endonuclease DNA fingerprints of the viruses isolated from myeloencephalitis and abortion were indistinguishable by Bam HI but were distinguishable using Bgl I, Pvu II, Xho I and Hind III. The restriction endonuclease DNA fingerprints of 3 EHV1 strains known to cause myeloencephalitis were compared with each other and with EHV1 strains not known to be associated with myeloencephalitis. The Bgl I Pvu II and Hind III DNA fingerprints of the 3 myeloencephalogenic strains appear distinguishable from non-myeloencephalogenic strains. Abortion was induced in a mare by intrauterine inoculation of EHV4. The Bam HI, Bgl I, Pvu II, Xho I and Hind III restriction endonuclease DNA fingerprints of the inoculum virus were indistinguishable from virus recovered from the foetus. It was concluded that passage of the virus through the foetus did not detectably alter the restriction endonuclease DNA fingerprint.

[Several molecular biologic characteristics of herpes virus isolated from lymphoblastoid cell lines from pavian hamadrils with malignant lymphoma]. / Nekotorye molekuliarno-biologicheskie kharakteristiki virusa gerpesa, izolirovannogo iz limfoblastoidnykh kletochnykh linii ot pavianov gamadrilov so zlokachestvennoi limfomoi

Cells of KMPG-1 and SPG-2 cultures obtained from the bone marrow and spleen of baboon hamadryads with hemoblastosis produce herpes-like virus (HLV) having the density of 1.21 and 1.27 g/cm3 in sucrose and cesium chloride density gradients. The density of nucleocapsids is 1.26 and 1.30 g/cm3 correspondingly. The HLV DNA sedimentation constant- 54 +/- 3S, and its density 1.717 g/cm3. The sera of patients with Burkitt's lymphoma and nasopharyngeal carcinoma can bind H3-thymidine labelled HLV that evidences a close antigenic relationship between HLV and NV virus.