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Isolation and characterisation of a ruminant alphaherpesvirus closely related to bovine herpesvirus 1 in a free-ranging red deer.

Thiry J, Widén F, Grégoire F, Linden A, Belák S, Thiry E - BMC Vet. Res. (2007)

Bottom Line: Besides the close relationship between all ruminant alphaherpesviruses, the phylogenetic analysis revealed that the isolate clustered with CvHV-1.Data demonstrate that a CvHV-1 strain, named Anlier, circulates in wild red deer in continental Europe.All together, these results improve our understanding of ruminant alphaherpesviruses.

View Article: PubMed Central - HTML - PubMed

Affiliation: Virology and Viral Diseases, Department of Infectious and Parasitic Diseases, Faculty of Veterinary Medicine, University of Liège, B-4000 Liège, Belgium. julien.thiry@ulg.ac.be

ABSTRACT

Background: The genus Varicellovirus of the Herpesviridae subfamily Alphaherpesvirinae includes a cluster of viruses antigenically and genetically related to bovine herpesvirus 1 (BoHV-1): namely bovine herpesvirus 5 (BoHV-5), bubaline herpesvirus 1 (BuHV-1), caprine herpesvirus 1 (CpHV-1), cervid herpesviruses 1 (CvHV-1) and 2 (CvHV-2) and elk herpesvirus 1 (ElkHV-1). Considering the serological relationship between these ruminant alphaherpesviruses, several surveys have studied the occurrence of BoHV-1 related virus infection in wild and domestic ruminant species. In this way, a recent investigation has indicated, in Belgium, a high increase in the serological prevalence of BoHV-1 related virus infection in free-ranging red deer population. In this context, it has been decided to investigate the presence of an alphaherpesvirus spreading in the Belgian free-ranging red deer population.

Results: The current study reports the first isolation in a free-ranging red deer of a BoHV-1 closely related virus. The isolate was antigenically, genomically and genetically characterised by comparison with several ruminant alphaherpesvirus. Immunofluorescence assays revealed the isolate was antigenically distinct from bovine and caprine alphaherpesviruses. Similarly, BamHI and BstEII restriction analyses demonstrated the genomic difference between the isolate and the other ruminant alphaherpesviruses. Next, the sequencing of selected parts of UL27 and US8 genes showed a high degree of homologies between each BoHV-1 related ruminant alphaherpesvirus and the isolate. Besides the close relationship between all ruminant alphaherpesviruses, the phylogenetic analysis revealed that the isolate clustered with CvHV-1.

Conclusion: The first isolation of a virus closely related to BoHV-1 in a free-ranging red deer is reported. Data demonstrate that a CvHV-1 strain, named Anlier, circulates in wild red deer in continental Europe. Anlier strain show consistent differences with the virus isolated from Scottish farmed red deer. All together, these results improve our understanding of ruminant alphaherpesviruses.

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Indirect immunofluorescence staining of MDBK cells infected with either BoHV-1, BoHV-5, CpHV-1, CvHV-1, CvHV-2, or the Anlier isolate. Cells were incubated until viral plaques appeared and were then treated as described in Materials and Methods. 6C3 is the primary antibody and was detected by FITC-conjugated rabbit immunoglobulin anti-mouse IgG. Presence of viral plaques was checked by transmission microscopy before epifluorescence microscopy. All immunofluorescence stainings were performed three times. Symbols: +, positive signal; -, negative signal; +/-, weak signal.
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Figure 1: Indirect immunofluorescence staining of MDBK cells infected with either BoHV-1, BoHV-5, CpHV-1, CvHV-1, CvHV-2, or the Anlier isolate. Cells were incubated until viral plaques appeared and were then treated as described in Materials and Methods. 6C3 is the primary antibody and was detected by FITC-conjugated rabbit immunoglobulin anti-mouse IgG. Presence of viral plaques was checked by transmission microscopy before epifluorescence microscopy. All immunofluorescence stainings were performed three times. Symbols: +, positive signal; -, negative signal; +/-, weak signal.

Mentions: In order to discriminate antigenically the Anlier isolate between bovine, caprine and cervid alphaherpesviruses, five selected monoclonal antibodies were tested by immunofluorescence assay on cells infected separately with the five related alphaherpesviruses: G14G11F5 for BoHV-1, 2915 for BoHV-5, 2E5G5G1 for CpHV-1, 6C2 for CvHV-1 and 5G10 for CvHV-2. Monoclonal antibodies detected their specific viruses as demonstrated by Keuser and collaborators [29], however, none of them reacted with the Anlier isolate. One can speculate that since CvHV-1 was also isolated from a red deer, CvHV-1 and the Anlier isolate might cross-react serologically. Therefore, other monoclonal antibodies were assessed to detect specifically the Anlier isolate. One monoclonal antibody, 6C3, identified the Anlier isolate. However, a weak reaction was also observed on CvHV-1 or CvHV-2 infected cells (Figure 1). Taken together, these results demonstrated that the Anlier isolate was antigenically distinct from BoHV-1, BoHV-5 and CpHV-1, but presumably possesses some common epitopes with CvHV-1 and CvHV-2.


Isolation and characterisation of a ruminant alphaherpesvirus closely related to bovine herpesvirus 1 in a free-ranging red deer.

Thiry J, Widén F, Grégoire F, Linden A, Belák S, Thiry E - BMC Vet. Res. (2007)

Indirect immunofluorescence staining of MDBK cells infected with either BoHV-1, BoHV-5, CpHV-1, CvHV-1, CvHV-2, or the Anlier isolate. Cells were incubated until viral plaques appeared and were then treated as described in Materials and Methods. 6C3 is the primary antibody and was detected by FITC-conjugated rabbit immunoglobulin anti-mouse IgG. Presence of viral plaques was checked by transmission microscopy before epifluorescence microscopy. All immunofluorescence stainings were performed three times. Symbols: +, positive signal; -, negative signal; +/-, weak signal.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2194762&req=5

Figure 1: Indirect immunofluorescence staining of MDBK cells infected with either BoHV-1, BoHV-5, CpHV-1, CvHV-1, CvHV-2, or the Anlier isolate. Cells were incubated until viral plaques appeared and were then treated as described in Materials and Methods. 6C3 is the primary antibody and was detected by FITC-conjugated rabbit immunoglobulin anti-mouse IgG. Presence of viral plaques was checked by transmission microscopy before epifluorescence microscopy. All immunofluorescence stainings were performed three times. Symbols: +, positive signal; -, negative signal; +/-, weak signal.
Mentions: In order to discriminate antigenically the Anlier isolate between bovine, caprine and cervid alphaherpesviruses, five selected monoclonal antibodies were tested by immunofluorescence assay on cells infected separately with the five related alphaherpesviruses: G14G11F5 for BoHV-1, 2915 for BoHV-5, 2E5G5G1 for CpHV-1, 6C2 for CvHV-1 and 5G10 for CvHV-2. Monoclonal antibodies detected their specific viruses as demonstrated by Keuser and collaborators [29], however, none of them reacted with the Anlier isolate. One can speculate that since CvHV-1 was also isolated from a red deer, CvHV-1 and the Anlier isolate might cross-react serologically. Therefore, other monoclonal antibodies were assessed to detect specifically the Anlier isolate. One monoclonal antibody, 6C3, identified the Anlier isolate. However, a weak reaction was also observed on CvHV-1 or CvHV-2 infected cells (Figure 1). Taken together, these results demonstrated that the Anlier isolate was antigenically distinct from BoHV-1, BoHV-5 and CpHV-1, but presumably possesses some common epitopes with CvHV-1 and CvHV-2.

Bottom Line: Besides the close relationship between all ruminant alphaherpesviruses, the phylogenetic analysis revealed that the isolate clustered with CvHV-1.Data demonstrate that a CvHV-1 strain, named Anlier, circulates in wild red deer in continental Europe.All together, these results improve our understanding of ruminant alphaherpesviruses.

View Article: PubMed Central - HTML - PubMed

Affiliation: Virology and Viral Diseases, Department of Infectious and Parasitic Diseases, Faculty of Veterinary Medicine, University of Liège, B-4000 Liège, Belgium. julien.thiry@ulg.ac.be

ABSTRACT

Background: The genus Varicellovirus of the Herpesviridae subfamily Alphaherpesvirinae includes a cluster of viruses antigenically and genetically related to bovine herpesvirus 1 (BoHV-1): namely bovine herpesvirus 5 (BoHV-5), bubaline herpesvirus 1 (BuHV-1), caprine herpesvirus 1 (CpHV-1), cervid herpesviruses 1 (CvHV-1) and 2 (CvHV-2) and elk herpesvirus 1 (ElkHV-1). Considering the serological relationship between these ruminant alphaherpesviruses, several surveys have studied the occurrence of BoHV-1 related virus infection in wild and domestic ruminant species. In this way, a recent investigation has indicated, in Belgium, a high increase in the serological prevalence of BoHV-1 related virus infection in free-ranging red deer population. In this context, it has been decided to investigate the presence of an alphaherpesvirus spreading in the Belgian free-ranging red deer population.

Results: The current study reports the first isolation in a free-ranging red deer of a BoHV-1 closely related virus. The isolate was antigenically, genomically and genetically characterised by comparison with several ruminant alphaherpesvirus. Immunofluorescence assays revealed the isolate was antigenically distinct from bovine and caprine alphaherpesviruses. Similarly, BamHI and BstEII restriction analyses demonstrated the genomic difference between the isolate and the other ruminant alphaherpesviruses. Next, the sequencing of selected parts of UL27 and US8 genes showed a high degree of homologies between each BoHV-1 related ruminant alphaherpesvirus and the isolate. Besides the close relationship between all ruminant alphaherpesviruses, the phylogenetic analysis revealed that the isolate clustered with CvHV-1.

Conclusion: The first isolation of a virus closely related to BoHV-1 in a free-ranging red deer is reported. Data demonstrate that a CvHV-1 strain, named Anlier, circulates in wild red deer in continental Europe. Anlier strain show consistent differences with the virus isolated from Scottish farmed red deer. All together, these results improve our understanding of ruminant alphaherpesviruses.

Show MeSH
Related in: MedlinePlus