Partial characterization of a novel gammaherpesvirus isolated from a European badger (Meles meles)

Abstract

A herpesvirus causing a cytopathic effect was isolated from pulmonary fibroblast cultures established from a European badger (Meles meles).A study was undertaken to classify and to assess some in-vitro growth characteristics of this virus. From a panel of 27 mammalian cell lines, in-vitro replication of the badger herpesvirus (BadHV) was only demonstrated with a mink lung cell line, suggesting a high degree of host specificity. Using PCR with degenerate primers, three independent fragments of the BadHV genome were sequenced. The largest of these fragments comprised a 6n2 kb segment including the DNA polymerase and glycoprotein B genes. Phylogenetic analysis of these sequences demonstrated that the BadHV is novel and clearly grouped with members of the Gammaherpesvirinae. In view of the oncogenic and immunosuppressive potential of many related herpesviruses, it is possible that BadHV can impact on existing acute or chronic disease in badgers.This report describes the partial characterization of a herpesvirus isolated from a European badger (Meles meles). The cytopathic effect (CPE) caused by this virus appeared in primary pulmonary fibroblast cultures established during the course of studies to evaluate a candidate DNA vaccine for Mycobacterium bovis (Chambers et al., 2001). These cultures were established from tissues collected as part of a statutory removal operation from a young adult female badger from Cornwall, South-West England, in 1996. At the time of presentation, this badger was in a poor nutritional state, weighing 6n0 kg. Post-mortem examination revealed a few scattered, 1 mm-diameter, white foci in the liver parenchyma and in the cortex of each kidney. Poorly defined 0n5 mm lesions were also observed in each lobe of the lungs. Histological examination of the lungs revealed multiple eosinophilic granulomas and diffuse inflammatory changes with eosinophils as the prominent cell type ; the changes were of the type observed in verminous pneumonia. In the kidneys and liver, there were multifocal inflammatory mononuclear cell infiltrates in the interstitium. However, no inclusion bodies or other histological changes characteristic of virus infection were observed. Cultural, histopathological and immunological tests for the diagnosis of M. bovis infection proved negative.After harvesting alveolar macrophages from lungs, a portion of the apical lobe was removed and comminuted and cells were dispersed using trypsin prior to seeding onto plastic culture flasks as described by Chambers et al. (2001). Once confluent, cultures were passaged by splitting at a ratio of 1 : 2. These cells were examined microscopically on alternate days. On the fourth passage, a CPE resembling that of a herpesvirus was observed. Examination of the infected cells by electron microscopy confirmed the presence of enveloped, herpesviruslike particles with a capsid diameter of approximately 105 nm. Since there are no published reports of a herpesvirus in this species, a more detailed investigation into the phenotypic characteristics and m...

Novel Gammaherpesviruses in North American Domestic Cats, Bobcats, and Pumas: Identification, Prevalence, and Risk Factors

“…In our initial analysis, 4 of 60 domestic cat samples were PCR positive, and 453-bp gB sequences amplified from these samples were identical ( Table 2). Comparison of this sequence to sequences in GenBank by BLAST analysis demonstrated homology to GHV gB sequences from other species, with the greatest similarity (80% nucleotide identity) being to mustelid herpesvirus 1 (MusHV1) of European badgers (36). The presence of this novel GHV sequence in multiple cats provided evidence for the likely existence of a novel GHV in cats.…”

Detection of Mustelid Herpesvirus-1 Infected European Badgers (Meles meles) in the British Isles

“…Mustelid herpesvirus-1 (MusHV-1) is a newly described herpesvirus isolated from a primary pulmonary cell culture derived from a European badger (Meles meles) in Cornwall, England (Banks et al, 2002). Phylogenetic analysis of sequence data demonstrates that MusHV-1 is a novel member of the Rhadinovirus genus within the Gammaherpesvirinae closely related to equine herpesvirus-2 and -5.…”

Identification of a novel herpesvirus associated with cutaneous ulcers in a fisher (Martes pennanti)

“…2,4,7,8 The fetal mink lung (ML) epithelial cell line g was also used because it is derived from a member of the Mustelidae family. 1 Ten-day-old embryonated eggs h (inoculated into the chorioallantoic sac and the chorioallantoic membrane) were also used for virus isolation.…”

“…Nucleotide comparison showed that the nearest nt homology was with the viral DNA polymerase (DPOL) gene of Mustelid herpesvirus 1 (MusHV-1; also known as Badger herpesvirus; order Herpesvirales, family Herpesviridae, subfamily Gammaherpesvirinae, genus Percavirus). 1 The Fisher herpesvirus (FiHV) partial sequence possesses 85.3% nt identity and 91.7% aa identity with the MusHV-1 DPOL gene published sequence (Fig. 3).…”

“…The novel viral DNA sequence of FiHV was most closely related to the badger herpesvirus (i.e., MusHV-1). 1 Like the fisher, the badger is an animal classified in the Mustelidae family. 15 Thus, it is reasonable to assume that FiHV is a new herpesvirus classified in the subfamily Gammaherpesvirinae.…”