The detection and phylogenetic analysis of equine herpesviruses 1, 4 and 5 identified in nasal swab samples of asymptomatic horses from Serbia and Bosnia and Herzegovina


Asymptomatic horses
EHV‑5Phylogenetic analysis

How to Cite

Radalj, A., Milic, N., Stevanovic, O., & Nisavic, J. (2021). The detection and phylogenetic analysis of equine herpesviruses 1, 4 and 5 identified in nasal swab samples of asymptomatic horses from Serbia and Bosnia and Herzegovina. Veterinaria Italiana, 57(4), 265–274. https://doi.org/10.12834/VetIt.1767.9329.3


Nasal swabs originating from 112 apparently clinically healthy and unvaccinated horses of different age, breed and from diverse rearing conditions from Serbia and Bosnia and Herzegovina were examined for the presence of equine herpesviruses 1, 4 and 5 using multiplex nested PCR (Mn‑PCR) and virus isolation. The detected viruses were subsequently characterised by gB gene nucleotide sequencing and their phylogenetic analysis was performed. The infections with EHV‑1, EHV‑4, and EHV‑5 in the examined horse populations are apparently chronic, subclinical and persistent, whilst the shedding of EHV‑1 and EHV‑5 was confirmed by their successful isolation. A connection was established between the finding of EHVs and rearing conditions since horses kept together in stables were positive for at least one EHV in contrast to animals held free grazing or individually. EHV‑5 was found most often in younger horses, however descending in frequency in animals up to 10 years of age. The phylogenetic analysis showed that the identified EHV strains group mostly with Turkish and German strains of respective viruses. A certain degree of genetic heterogeneity was determined regarding the identified EHV‑5 strains in contrast to EHV‑1 and EHV‑4.


Ataseven V.S., Bilge Dagalp S., Oguzoglu T.C., Karapinar Z., Guzel M. & Tan M.T. 2010. Detection and sequence analysis of equine gammaherpesviruses from horses with respiratory tract disease in Turkey. Transbound Emerg Dis, 57, 271–276.

Ataseven V.S, Oguzoglu T.C., Dincer E. & Dagalp S.B. 2016. Partial sequence of the gB gene of equid herpesvirus type 1 isolates associated with abortion in Turkey. Ankara Univ Vet Fak Derg, 63, 277-281.

Bell S.A., Balasuriya U.B., Gardner I.A., Barry P.A., Wilson W.D., Ferraro G.L. & MacLachlan N.J. 2006. Temporal detection of equine herpesvirus infections of a cohort of mares and their foals. Vet Microbiol, 116, 249–257.

Bilge Dagalp S., Babaoglu A.R., Ataseven V.S., Karapinar Z., Timurkan M.O., Dogan F., Ozkul A. & Alkan F. 2018. Determination of presence of equid alpha and gammaherpesvirus infections in foals with respiratory distress. Ankara Univ Vet Fak Derg, 65, 63-68.

Brault S.A. & MacLachlan N.J. 2011. Equid gammaherpesviruses: persistent bystanders or true pathogens? Vet J, 187, 14–15.

Davison A.J., Eberle R., Ehlers B., Hayward G.S., McGeoch D.J., Minson A.C., Pellett P.E., Roizman B., Studdert M.J. & Thiry E. 2009. The order Herpesvirales. Arch Virol, 154, 171-177.

Diallo I.S., Hewitson G.R., de Jong A., Kelly M.A., Wright D.J., Corney B.G. & Rodwell B.J. 2008. Equine herpesvirus infections in yearlings in South-East Queensland. Arch Virol, 153, 1643-1649.

Dunowska M., Holloway S.A., Wilks C.R. & Meers J. 2000. Genomic variability of equine herpesvirus-5. Arch Virol, 145, 1359–1371.

Dunowska M., Wilks C.R., Studdert M.J. & Meers J. 2002. Equine respiratory viruses in foals in New Zealand. N Z Vet J, 50, 140–147.

Edington N., Welch H.M. & Griffiths L. 1994. The prevalence of latent Equid herpesviruses in the tissues of 40 abattoir horses. Equine Vet J, 26, 140–142.

Foote C.E., Love D.N., Gilkerson J.R. & Whalley J.M. 2004. Detection of EHV-1 and EHV-4 DNA in unweaned Thoroughbred foals from vaccinated mares on a large stud farm. Equine Vet J, 36, 341-345.

Gilkerson J.R., Whalley J.M. & Drummer H.E. 1999. Epidemiological studies of equine herpesvirus 1 (EHV-1) in thoroughbred foals: a review of studies conducted in the Hunter Valley of New South Wales between 1995 and 1997. Vet Microbiol, 68, 15-25.

Hartley C.A., Dynon K.J., Mekuria Z.H., El-Hage C.M., Holloway S.A. & Gilkerson J.R. 2013. Equine gammaherpesviruses: perfect parasites? Vet Microbiol, 167, 86-92.

Heldens J.G., Hannant D., Cullinane A.A., Prendergast M.J., Mumford J.A., Nelly M., Kydd J.H., Weststrate M.W. & van den Hoven R. 2001. Clinical and virological evaluation of the efficacy of an inactivated EHV1 and EHV4 whole virus vaccine (Duvaxyn EHV1,4). Vaccination/challenge experiments in foals and pregnant mares. Vaccine, 19, 4307-4317.

Laabassi F., Hue E., Fortier C., Morilland E., Legrand L., Hans A. & Pronost S. 2017. Epidemiology and molecular detection of equine herpesviruses in western Algeria in 2011. Vet Microbiol, 207, 205–209.

Marenzoni M.L., Coppola G., Maranesi M., Passamonti F., Cappelli K., Capomaccio S., Verini Supplizi A., Thiry E. & Coletti M. 2010. Age-dependent prevalence of equid herpesvirus 5 infection. Vet Res Commun, 34, 703–708.

Milić N., Radalj A. & Nišavić J. 2018. Standard and molecular methods in the diagnostics of infections caused by equine herpesviruses 1 and 4. Vet Glasnik, 72, 68-79.

Negussie H., Gizaw D., Tesfaw L., Li Y., Oguma K., Sentsui H., Tessema T.S. & Nauwynck H.J. 2017. Detection of Equine Herpesvirus (EHV) -1, -2, -4 and -5 in Ethiopian Equids with and without Respiratory Problems and Genetic Characterization of EHV-2 and EHV-5 Strains. Transbound Emerg Dis, 64, 1970-1978.

Nordengrahn A., Merza M., Ros C., Lindholmc A., Palfl V., Hannant D. & Belak S. 2002. Prevalence of equine herpesvirus types 2 and 5 in horse populations by using type-specific PCR assays. Vet Res, 33, 251–259.

World Organisation for Animal Health (OIE). 2018. OIE Terrestrial Manual 2017. Chapter 2.5.9. Equine rhinopneumonitis (Infection with equid herpesvirus-1 and -4). (http://www.oie.int/fileadmin/Home/eng/Health_standards/tahm/2.05.09_EQUINE_RHINO.pdf/ accessed on 23 December 2018).

Patel J.R. & Heldens J. 2005. Equine herpesviruses 1 (EHV-1) and 4 (EHV-4) - Epidemiology, disease and immunoprophylaxis: A brief review. Vet J, 170, 14–23.

Ploszay G., Rola J., Larska M. & Zmudzinski J.F. 2013. First report on equine herpesvirus type 4 isolation in Poland-evaluation of diagnostic tools. Pol J Vet Sci, 16, 493–500.

Radalj A., Nišavić J., Krnjaić D., Valčić M., Jovanović T., Veljović Lj. & Milić N. 2018. Detection and molecular characterization of equine herpesviruses 1, 2, and 5 in horses in the Republic of Serbia. Acta Vet Brno, 87, 27-34.

Slater J. 2014. Equine Herpesviruses. In Equine Infectious Diseases, 2nd Ed. (D.C. Sellon & M.T. Long, eds). Saunders, Elsevier, St. Louis, 151-168.

Slater J.D., Borchers K., Thackray A.M. & Field H.J. 1994. The trigeminal ganglion is a location for equine herpesvirus 1 latency and reactivation in the horse. J Gen Virol, 75, 2007–2016.

Stasiak K., Dunowska M. & Rola J. 2018. Prevalence and sequence analysis of equid herpesviruses from the respiratory tract of Polish horses. Virol J, 15, 106.

Turan N., Yildirim F., Altan E., Sennazli G., Gurel A., Diallo I. & Yilmaz H. 2012. Molecular and pathological investigations of EHV-1 and EHV-4 infections in horses in Turkey. Res Vet Sci, 93, 1504–1507.

Van Maanen C. 2002. Equine herpesvirus 1 and 4 infections: An update. Vet Quart, 24, 57-78.

van Regenmortel M.H.V., Fauquet C.M., Bishop D.H.L., Carstens E.B., Estes M.K., Lemon S.M., Maniloff J., Mayo M.A., McGeoch D.J., Pringle C.R. & Wickner R.B. 2000. In Virus taxonomy: classification and nomenclature of viruses. Seventh report of the International Committee on Taxonomy of Viruses (M.H.V van Regenmortel, C.M. Fauquet, D.H.L. Bishop, E.B. Carstens, M.K. Estes, S.M. Lemon, J. Maniloff, M.A. Mayo, D.J. McGeoch, C.R. Pringle & R.B. Wickner, eds). Academic Press, San Diego.

Wang L., Raidal S.L., Pizzirani A. & Wilcox G.E. 2007. Detection of respiratory herpesviruses in foals and adult horses determined by nested multiplex PCR. Vet Microbiol, 121, 18–28.

Welch H.M., Bridges C.G., Lyon A.M., Griffiths L. & Edington N. 1992. Latent equid herpesviruses 1 and 4: Detection and distinction using the polymerase chain reaction and co-cultivation from lymphoid tissues. J Gen Virol, 73, 261–268.

Williams K.J., Robinson N.E., Lim A., Brandenberger C., Maes R., Behan A. & Bolin S.R. 2013. Experimental Induction of Pulmonary Fibrosis in Horses with the Gammaherpesvirus Equine Herpesvirus 5. PLoS One, 8, 1–15.

Copyright (c) 2022 Veterinaria Italiana