A Phylogenetic and Genotyping Study of Bovine Herpesvirus Type 4 (BHV-4) in Turkey

Infertility and reproductive infections are the huge problems for the diary management throughout the world. Bovine herpesviruses act an enormous role in these complicated problems. Bovine Herpesvirus Type 1 (BHV-1) is the most outstanding herpesvirus causing genital and uterine tracts infections among other reproductive viral agents, however, Bovine Herpesvirus Type 4 (BHV-4) is also responsible in terms of similar symptoms and diseases. The main aims of the study are both to investigate the underlying potential presence of BHV-4 in subclinical uterine tract infection, and both to perform molecular and recombination analyses. A herd including 25 repeat breeder cows were investigated by BHV-4. Two out of them were determined BHV-4 infected after a series of Polymerase Chain Reactions (PCRs) tests which able to amplify partial Glycoprotein B (gB) and Thymidine Kinase (TK) gene regions. Obtained sequences were analyzed by using phylogenetic and recombinational software, and two Maximum Likelihood (ML) tree have been constructed. To results, novel Turkish BHV-4 sequences fell into Genotype I in both constructed Maximum Likelihood (ML) phylogenetic trees, however, no recombination evidence has been observed in relevant software. This report is the first genotyping study of BHV-4 from Turkey. This study showed that Turkish BHV-4 substantially originated from European strains and might be observed in different clinical presentations. This suggests that BHV-4 should be deeply investigated by further molecular techniques and included in diagnostic panels for reproductive diseases viruses.

___

  • 1) Chastant-Maillard S. Impact of bovine herpesvirus 4 (BoHV-4) on reproduction. Transbound Emerg Dis. 2015;62(3):245–251. doi:10.1111/tbed.12155
  • 2) Bartha A, Juhasz M, Liebermann H. Isolation of a bovine herpesvirus from calves with respiratory disease and keratoconjunctivitis. A preliminary report. Acta Vet Acad Sci Hung. 1966;16(3):357–358.
  • 3) Davison AJ. Herpesvirus systematics. Vet Microbiol. 2010;143(1):52–69. doi:10.1016/j.vetmic.2010.02.014
  • 4) Bellino C, Iussich S, Biasato I, et al. Potential pathogenetic role of bovine herpesvirus 4 in two dairy cows with dermatitis-pyrexia-hemorrhagic syndrome. J Clin Microbiol. 2015;53(8):2763–7. doi:10.1128/JCM.00717-15
  • 5) Egyed L. Bovine herpesvirus type 4: a special herpesvirus (review article). Acta Vet Hung. 2000;48(4):501–13. doi:10.1556/004.48.2000.4.13
  • 6) Verna AE, Manrique JM, Perez SE, et al. Genomic analysis of bovine herpesvirus type 4 (BoHV-4) from Argentina: high genetic variability and novel phylogenetic groups. Vet Microbiol. 2012;160(1–2):1–8. doi:10.1016/j.vetmic.2012.04.039
  • 7) Gagnon CA, Traesel CK, Music N, et al. 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. 2017;4:92. doi:10.3389/fvets.2017.00092
  • 8) Zimmermann W, Broll H, Ehlers B, et al. Genome sequence of bovine herpesvirus 4, a bovine rhadinovirus, and identification of an origin of DNA replication. J Virol. 2001;75(3):1186–94. doi:10.1128/JVI.75.3.1186-1194.2001
  • 9) Rasheed MA, Ansari AR, Ihsan A, et al. Prediction of conserved sites and domains in glycoproteins B, C and D of herpes viruses. Microb Pathog. 2018;116:91-99.
  • 10) Morán P, Pérez S, Odeón A, et al. Comparative analysis of replicative properties of phylogenetically divergent, Argentinean BoHV-4 strains in cell lines from different origins. Comp Immunol Microbiol Infect Dis. 2019;63:97-103. doi:10.1016/j.cimid.2019.01.014
  • 11) Bilge-Dagalp S, Demir AB, Gungor E, et al. The seroprevalence of Bovine Herpes Virus Type 4 (BHV-4) infection in dairy herds in Turkey and possible interaction with reproductive disorders. Rev Med Vet (Toulouse). 2007;158(4):201.
  • 12) Bilge-Dagalp S, Gungor E, Demir AB, et al. The investigation of the presence of bovine herpesvirus type 4 (BoHV-4) in cows with metritis in a dairy herd. Ankara Univ Vet Fak Derg. 2010;57(2):87-91.
  • 13) Gür, S. Prevalence of bovine viral diarrhoea, bovine herpesvirus type 1 and 4 infections in repeat breeding cows in Western Turkey. Braz J Vet Res Anim Sci. 2011;48(3):228-233.
  • 14) Aslan ME, Azkur AK, Gazyagci S. Epidemiology and genetic characterization of BVDV, BHV-1, BHV-4, BHV-5 and Brucella spp. infections in cattle in Turkey. J Vet Med Sci. 2015;14-0657.
  • 15) Kale M, Ata A, Kocamüftüoglu M, et al. Bovine herpesvirus type 4 (BHV-4) infection in relation to fertility in repeat breeder dairy cows. Acta Vet (Beograd). 2011;61(1):13-19.
  • 16) Yavru S, Avci O, Kale M, et al. Serologic and virologic investigation of BHV-1, BVDV and BHV-4 in cattle with metritis. Anim Vet Sci. 2014;2(5):142-145.
  • 17) Yildirim Y, Yilmaz V, Kalaycioglu AT, et al. An investigation of a possible involvement of BVDV, BHV-1 and BHV-4 infections in abortion of dairy cattle in Kars district of Turkey. Kafkas Univ Vet Fak Derg. 2011;17:879-883.
  • 18) Chomczynski P, Sacchi N. The single-step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction: twenty-something years on. Nat Protoc. 2006;1(2):581.
  • 19) Wellenberg GJ, Verstraten ERAM, Belak S, et al. Detection of bovine herpesvirus 4 glycoprotein B and thymidine kinase DNA by PCR assays in bovine milk. J Virol Meth. 2001;97(1-2):101-112.
  • 20) Kumar S, Stecher G, Li M, et al. MEGA X: molecular evolutionary genetics analysis across computing platforms. Mol Bio Evol. 2018;35(6):1547-1549.
  • 21) Martin DP, Murrell B, Golden M, et al. RDP4: Detection and analysis of recombination patterns in virus genomes. Virus Evol. 2015;1(1).
  • 22) Areda D, Chigerwe M, Crossley B. Bovine herpes virus type-4 infection among postpartum dairy cows in California: risk factors and phylogenetic analysis. Epidemiol Infect. 2018;146(7):904-912.