Genetic variability of bovine herpesvirus type 4 (BoHV-4) field strains from Turkish cattle herds

Supplementary Files

Table I
Figure 1
Figure 2
Figure 3
Figure 4
Table II
Table III
Figure 5
Figure 6
Figure 7


Bovine Herpesvirus Type 4
glycoprotein B (gB)
thymidine kinase (TK)
Virus isolation

How to Cite

Bilge Dağalp, S., Dogan, F., Babaoglu, A. R., Aligholipour Farzani, T., & Alkan, F. (2021). Genetic variability of bovine herpesvirus type 4 (BoHV-4) field strains from Turkish cattle herds. Veterinaria Italiana, 57(1), 49–59. https://doi.org/10.12834/VetIt.2095.11150.1


Bovine herpesvirus type 4 (BoHV‑4) is a common virus in the world that is detected in clinically ill or in apparently healthy cattle. This study provides a molecular characterization of BoHV‑4 strains from 24 cattle some showing respiratory and/or reproductive problems and some without any apparent clinical sign. This study also reported the growth properties of five BoHV‑4 field isolates. The 24 sampled cattle came from 13 different herds in 10 provinces collected between 2007 and 2018. Phylogenetic analysis using partially amplified nucleotide sequences of ORF8 genes coding glycoprotein B (n = 24) and ORF3 genes coding thymidine kinase (n = 9), demonstrated genetic variability among the BoHV‑4 strains analysed. The partial gB gene sequences clustered in three different genotypes (genotype I, II and III) were located within the genotype I cluster, such as Movar strain. The analysis of the five BoHV‑4 strains isolated from vaginal swabs (n = 2), nasal swab (n = 1), and brain samples (n = 2) revealed no significant differences in their growth properties in MDBK cell culture.



Altamiranda E.G., Manrique J.M., Pérez S.E., Ríos G.L., Odeón A.C., Leunda M.R. & Verna A. 2015. Molecular characterization of the first bovine herpesvirus 4 (BoHV-4) strains isolated from in vitro bovine embryos production in Argentina. PloS One, 10(7), e0132212.

Altschul S.F., Madden T.L., Schäffer A.A., Zhang J., Zhang Z., Miller W. & Lipman D.J. 1997. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res, 25(17), 3389-3402.

Areda D., Chigerwe M. & Crossley B. 2018. Bovine herpes virus type-4 infection among postpartum dairy cows in California: risk factors and phylogenetic analysis. Epidemiol Infect, 146(7), 904-912.

Bellino C., Iussich S., Biasato I., Peletto S., Caruso C., Gianella P., Cagnasso A. & D’Angelo A. 2015. Potential Pathogenetic Role of Bovine Herpesvirus 4 in Two Dairy Cows with Dermatitis-Pyrexia-Hemorrhagic Syndrome. J Clin Microbiol, 53 (8), 2763-2767.

Bilge Dağalp S., Alkan F., Çalışkan E., Yıldırım Y., Oğuzoğlu Ç. & Burgu İ.2012. The investigation of the herpesviruses (BoHV-1 and BoHV-4) on the occurrence of the reproductive disorders in dairy cattle herds, Turkey. Rev Med Vet, 163 (4), 206-211.

Bilge Dağalp S., Güngör E., Demir A.B. & Alkan F. 2007. The seroprevalence of Bovine Herpes Virus Type 4 (BHV4) infection in dairy herds in Turkey and possible interaction with reproductive disorders. Rev Med Vet, 158 (4), 201-205.

Bilge Dağalp S., Güngör E., Demir A.B., Pınar-Muz D., Yılmaz V., Oğuzoğlu T.Ç., Ataseven V.S. & Alkan F. 2011. The investigation of the presence of bovine herpesvirus type 4 (BoHV-4) in cows with metritis in a dairy herd. Vet J Ankara Univ, 57(2), 87-91.

Bublot M., Van Bressem M.F., Thiry E., Dubuisson J. & Pastoret P.P. 1990. Bovine herpesvirus 4 genome: cloning, mapping and strain variation analysis. J Gen Virol, 71, 133-142.

Campos F.S., Franco A.C., Oliveira M.T., Firpo R., Strelczuk G., Fontoura F.E., Kulmann M.I., Maidana S., Romera S.A., Spilki F.R., Silva A.D., Hübner S.O. & Roehe P.M. 2014. Detection of bovine herpesvirus 2 and bovine herpesvirus 4 DNA in trigeminal ganglia of naturally infected cattle by polymerase chain reaction. Vet Microbiol, 171, 182–188. .

Egyed J., Ballagi-Pordany A., Bartha A. & Belak S. 1996. Studies of in vivo distribution of bovine herpesvirus type 4 in the natural host. J Clin Microbiol, 34, 1091-1095.

Fichtelova V. & Kovarcik K. 2010. Characterization of two BHV-4 strains isolated in the Czech Republic. Vet Med (Praha), 55(3), 106-112.

Frazier K., Baldwin C.A., Pence M., West J., Bernard J., Liggett A., Miller D. & Hines M. 2002. Seroprevalence and comparison of isolates of endometriotropic bovine herpesvirus-4. J Vet Diagn Invest, 14, 457-462.

Gagnon C.A., Traesel C.K., Music N., Laroche J., Tison N., Auger J.P., Music S., Provost C., Bellehumeur C., Abrahamyan L., Carman S., Côteaux L. & Charette S.J. 2017. Whole genome sequencing of a Canadian Bovine gammaherpesvirus 4 strain and the Possible link between the Viral infection and respiratory and reproductive clinical Manifestations in Dairy cattle. Front Vet Sci, 4, 1-12.

Goltz M., Broll H., Mankertz A., Weigelt W., Ludwig H., Buhk H.J. & Borchers K. 1994. Glycoprotein B of bovine herpesvirus type 4: its phylogenetic relationship to gB equivalents of the herpesviruses. Virus Genes, 9, 53-59.

Gur S. & Dogan N. 2010. The possible role of bovine herpesvirus type-4 infection in cow infertility. Anim Sci J, 81, 304– 308.

Hall T.A. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series, 41, 95-98.

Houe H. 1995. Epidemiology of bovine viral diarrhea virus. Vet Clin North Am Food Anim Pract, 11 (3), 521-547.

Kale M., Ata A., Kocamüftüoglu M. & Hasırcıoglu S. 2011. Bovıne Herpesvırus Type 4 (BHV-4) infectıon in relatıon to fertility in repeat breeder daıry cows. Acta Vet (Beograd), 61, 13-19.

Kit S., Kit M., Ichimura H., Crandell R. & McConnel S. 1986. Induction of thymidine kinase activity by viruses with group B DNA genomes; bovine cytomegalovirus (bovine herpesvirus 4). Virus Res, 4, 197–212.

Lin T.M., Shi G.Y., Tsai C.F., Su H.I., Guo Y.I. & Wu H.I. 1997. Susceptibility of endotelial cells to bovine herpesvirus type 4 (BHV4). J Virol Methods, 63, 219-225.

Lomonte P., Bublot M., Pastoret P.P. & Thiry E. 1992. Location and characterization of the bovine herpesvirus type 4 thymidine kinase gene; comparison with thymidine kinase genes of other herpesviruses. Arch Virol, 127(1-4), 327-337.

Nettleton P.F., Herring J.A. & Herring A.J. 1983. Evaluation of an immunofluorescent test for the rapid diagnosis of field infections bovine rhinotracheitis. Vet Rec, 112, 298-300.

Pérez S.E., Verna A.E., Leunda M.R., Favier P.A., Ceriani M.C., Morán P.E. & Esteban E.N. 2011. Occurrence of bovine herpesvirus type 4 DNA in Argentinean Holstein cattle from Santiago del Estero, Argentina. Braz J Vet Res Anim Sci, 48(6), 454-463.

Reed L.J. & Muench H. 1938. A simple method of estimating fifty per cent endpoints. Am J Epidemiol, 27, 493–497.

Roizman B., Desroisers R.C., Fleckenstein B., Lopez C., Minson A.C. & Studdert M.J. 1992. The family herpesviridae: an update. Arch Virol, 123, 425-449.

Sambrook J., Fritsch E.F. & Maniatis T. 1989. Molecular Cloning : A Laboratory Manual. 2 Ed. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.

Tamura K., Dudley J., Nei M. & Kumar S. 2011. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol, 28, 2731–2739.

Thiry E., Bublot M., Dubuisson J., Van Biessem M.F., Lequarre A.S., Lomante P., Vanderplasschen A. & Pastoret P.P. Molecular biology of BHV4. Vet Microbiol, 33, 79-92.

Verna A. E., Manrique J. M., Pérez S. E., Leunda M. R., Pereyra S. B., Jones L. R. & Odeón A.C. 2012. Genomic analysis of bovine herpesvirus type 4 (BoHV-4) from Argentina: high genetic variability and novel phylogenetic groups. Vet Microbiol, 160, 1-8.

Verna A.E., Pérez S.E., Manrique J.M., Leunda M.R., Odeón A.C. & Jones L.R. 2016. Comparative study on the in vitro replication and genomic variability of Argentinean field isolates of bovine herpesvirus type 4 (BoHV-4). Virus Genes, 52, 372- 378.

Wellemans G., Van Opdenbosch E. & Mammericks M. 1986. Experimental inoculation of Bovine Herpes Virus 4 (Strain LVR 140) in pregnant and nonpregnant cows. Ann Rech Vet, 17(1), 89-94.

Wellenberg G.J., Verstraten E.R.A.M., Belak S., Verschuren F.A.M., Rijsewijk R., Peshev R. & Van Oirschot J.T. 2001. Detection of bovine herpes virus 4 glycoprotein B and thymidine kinase DNA by PCR assays in bovine milk. J Virol Methods, 97, 101-112.

Zimmerman W., Broll H., Ehlers B., Buhk H.J., Rosenthal A. & Goltz M. 2001. Genome sequence of bovine herpesvirus 4, a bovine Rhadinovirus, and identification of an origin of DNA replication. J Virol, 75, 1186-1194.