doi: 10.15389/agrobiology.2016.6.861eng

UDC 636.32/38:619:578.821.6



D.V. Yanzhieva1, N.I. Sal’nikov1, T.R. Usadov1, S.P. Zhivoderov1,
L.K. Saryglar2, A.V. Lunitsin1

1All-Russian Institute of Veterinary Virology and Microbiology, Federal Agency of Scientific Organizations, 1, ul. Akademika Bakuleva, pos. Vol’ginskii, Petushinskii Region, Vladimir Province, 601125 Russia, e-mail,,,,;
2Tuvan State University, 36, ul. Lenina, Kyzyl, Tuva Republic, 667000 Russia,

Received October 4, 2016


Orf is infectious disease of sheep and goats, characterized by lesions of mucosa of the oral cavity, skin of lips, head, mammary glands and limbs. The causal agent (virus of contagious ecthyma — Orf virus) is a member of genus Parapoxvirus, subfamily Chordopoxvirinae, family Poxviridae. Genome of the Orf virus consists of linear double stranded DNA (138 kbp) and contains 132 open reading frames. Its antigenic structure is poorly understood, and strains are not serologically identical. That is why the investigation of novel isolates of Orf virus is actual. However, we are not aware of such research in Russia possibly due to rare outbreaks recognized. We first identified Orf virus in sheep from Tuva Republic and studied biological properties of the isolate. Disease of sheep, characterized by erythema, vesicles, pustules and scabs on ears and legs has been registered in August 2015 in Tuva Republic. The scabs were sampled for isolation and identification of the causative agent. For this result three lambs were experimentally infected with the scab suspension. The mucocutaneous borders of the lips along the labial commissures were scarified and inoculum was loaded by cotton swabs. The lip vesicles appeared at days 4-5 and then formed pustules and scabs. After 2 weeks the lesions healing occurred. The suspension of scabs from lips of infected lambs was used to inoculate sheep kidney cell culture. In 7 days post inoculation the cell monolayer was harvested for second passage. However, we did not observe specific cytopathic effect in the monolayers during these passages. The skin swabs from experimentally infected lambs were examined by Real-Time PCR in accordance to the protocol developed by G. Venkatesan et al. (2014), and Orf virus DNA was detected at days 7 to 28 post infection. We also detected Orf virus DNA in scabs from the infected lambs and in lysates of the monolayers harvested after second passage. The PCR test was positive for reference strain IA82 of Orf virus unlike other viruses (nodular dermatitis vaccine strain, sheep pox strains Mongolian and B5/96 or goatpox strain QA/A-04) that confirmed specificity of the PCR system. Its analytical sensitivity to detect Orf virus genomic DNA was 1.3±0.03 lg TCD50/cm3. Thus, the pathogen which caused the disease of sheep in Tuva Republic in 2015 was isolated from experimentally infected lambs using sheep kidney cell culture and identified as Orf virus.

Keywords: sheep, Orf virus, PCR, cell culture.


Full article (Rus)

Full text (Eng)



  1. Kolomytsev A.A., Zakutskii N.I., Guzalova A.G., Mikolaichuk S.V., Morgunov Yu.P.  Veterinarnaya patologiya, 2008, 4: 31-37 (in Russ.).
  2. Delhon G., Tulman E.R., Afonso C.L., Lu Z., de la Concha-Bermejillo A., Lehmkuhl H.D.,  Piccone M.E., Kutish G.F., Rock D.L. Genomes of the parapoxviruses Orf virus and bovine papular stomatitis virus. J. Virol., 2004, 78(1): 168-177 CrossRef
  3. Bayindir Y., Bayraktar M., Karadag N., Ozcan H., Kayabas U., Otlu B., Durmaz R., Doganay M. Investigation and analysis of a human orf outbreak among people living on the same farm. New microbiologica, 2011, 34: 37-43.
  4. Hosamani A., Scagliarini A., Bhanuprakash V., McInnes C.J., Singh R.K. Orf: an update on current research and future perspectives. Expert Review on Antiinfective Therapy, 2009, 7(7): 879-893 CrossRef
  5. Bhanuprakash V., Venkatesan G., Balamurugan V., Hosamani M., Yogisharadhya R., Chauhan R.S., Pande A., Mondal B., Singh R.K. Pox outbreaks in sheep and goats at Makhdoom (Uttar Pradesh), India: evidence of sheep pox virus infection in goats. Transbound. Emerg. Dis., 2010, 57(5): 375-382 CrossRef
  6. Abubankr M.I., Abu-Elzein E.M., Housawi F.M., Abdelrahman A.O., Fadlallah M.E., Nayel M.N., Adam A.S., Moss S., Forrester N.L., Coloyan E., Gameet A., Al-Afaleq A.I., Gould E.A. Pseudo cow poxvirus: the etiological agent of contagious ecthyma (Auzdyk) in camels (Camelus dromedarius) in the Arabian peninsula. Vector Borne Zoonotic Diseases, 2007, 7(2): 257-260 CrossRef
  7. Abdelrabman K., Soliman H. Molecular and virological studies on contagious pustular dermatitis isolates from Egyption sheep and goats. Research in Veterinary Science, 2010, 89(2): 290-294 CrossRef
  8. Wise L.M., Savory L.J., Dryden N.H., Whelan E.M., Fleming S.B., Mercer A.A. The development of oral lesions in lambs naturally infected with orf virus. Veterinary Journal, 2007, 174(3): 663-664 CrossRef
  9. Venkatesan G., Balamurugan V., Bhanuprakash V. TaqMan based real-time duplex PCR for simultaneous detection and quantitation of Capri pox and Orf virus genomes in clinical samples. J. Virol. Methods, 2014, 201: 44-50 CrossRef
  10. Oem J.-K., Chung J.-Y., Kim Y.-J., Lee K.-K., Kim S.-H., Jung B.-Y., Hyun B.-H. Isolation and characterization of orf viruses from Korean black goats. J. Vet. Sci., 2013, 14(2): 227-230 CrossRef
  11. Bora D.P., Barman N.N., Das S.K., Bhanuprakash V., Yogisharadhya R., Venkatesan G., Kumar A., Rajbongshi G., Khatoon E., Chakraborty A., Bujarbaruah K.M. Identification and phylogenetic analysis of orf viruses isolated from outbreaks in goats of Assam, a northeastern state of India. Virus Genes, 2012, 45: 98-104 CrossRef
  12. Chan K.W., Lin J.W., Lee S.H., Liao C.J., Tsai M.C., Hsu W.L., Wong M.L., Shih H.C. Identification and phylogenetic analysis of orf virus from goats in Taiwan. Virus Genes, 2007, 35: 705-712 CrossRef
  13. Mondal B., Bera A.K., Hosamani M., Tembhurne P.A., Bandyopadhyay S.K. Detection of orf virus from an outbreak in goats and its genetic relation with other parapoxviruses. Veterinary Research Community, 2006, 30: 531-539 CrossRef
  14. Hosamani M., Yadav S., Kallesh D.J., Mondal B., Bhanuprakash V., Singh R.K. Isolation and characterization of an Indian orf virus from goats. Zoonoses Public Health, 2007, 54: 204-208 CrossRef
  15. Musser J.M.B., Taylor C.A., Guo J., Tizard I.R., Walker J.W. Development of a contagious ecthyma vaccine for goats. Am. J. Vet. Res., 2008, 69: 1366-1370 CrossRef
  16. Hosamani M., Bhanuprakash V., Scagliarini A., Singh R.K. Comparative sequence analysis of major envelope protein gene (B2L) of Indian orf viruses isolated from sheep and goats. Vet. Microbiol., 2006, 116: 317-324 CrossRef
  17. Venkatesan G., Balamurugan V., Bora D.P., Yogisharadhya R., Prabhu M., Bhanuprakash V. Sequence and phylogenetic analyses of an Indian isolate of orf virus from sheep. Veterinaria Italiana, 2011, 47(3): 323-332.
  18. Abrahão J.S., Campos R.K., Trindade G.S., Guedes M.I., Lobato Z.I., Mazur C., Ferreira P.C., Bonjardim C.A., Kroon E.G. Detection and phylogenetic analysis of orf virus from sheep in Brazil: a case report. Virol. J., 2009, 6: 47 CrossRef
  19. Chan K.W., Hsu W.L., Wang C.Y., Yang C.H., Lin F.Y., Chulakasian S., Wong M.L. Differential diagnosis of orf viruses by a single-step PCR. J. Virol. Methods, 2009, 160(1-2): 85-89 CrossRef
  20. Gallina L., Dal Pozzo F., Mc Innes C.J., Cardeti G., Guercio A., Battilani M., Ciulli S., Scagliarini A. A real-time PCR assay for the detection and quantification of orf virus. J. Virol. Methods, 2005, 134: 140-145 CrossRef
  21. Nitsche A., Buttner M., Wilhelm S., Pauli G. Meyer H. Real-Time PCR detection of parapoxvirus DNA. Clin. Chem., 2006, 52(2): 316-319 CrossRef
  22. Kottaridi C., Nomikou K., Lelli R., Markoulatos P., Mangana O. Laboratory diagnosis of contagious ecthyma: comparison of different PCR protocols with virus isolation in cell culture. J. Virol. Methods, 2006, 134: 119-124 CrossRef
  23. Venkatesan G., Balamurugan V., Bhanuprakash V. TaqMan based real-time duplex PCR for simultaneous detection and quantitation of Capri pox and Orf virus genomes in clinical samples. J. Virol. Methods, 2014, 201: 44-50 CrossRef
  24. Lakin G.F. Biometriya [Biometry]. Moscow, 1990 (in Russ.).