Keywords
Electron microscopy
ORF virus
PCR
Phylogenetic analysis
VLTF-1 gene
How to Cite
Abstract
Orf virus (ORFV; Family: Poxviridae) is the causative agent of contagious ecthyma, or Orf disease in sheep, goats and other domestic or wild ruminants with a worldwide distribution. The disease is endemic in Italy, but few data are available about its distribution and epidemiology. In the present study we analysed 32 clinical samples, obtained from crusted scab lesions of 5 goats and 27 sheep, from 19 suspected outbreaks of contagious ecthyma in Apulia and Basilicata regions between 2012 and 2014. Negative staining electron microscopy (EM) and polymerase chain reaction (PCR) targeting the late transcription factor gene (VLTF-1) were used to identify the virus. Isolation was also attempted on BHK-21 cell line. PCR was proved to be more sensitive than EM, as it detected the virus in 28 out of 32 samples, whereas the EM detected it only in 26 out of the 32 samples. The majority of isolated strains forms a monophyletic group; these isolates, according to the VLTF-1 sequencing, are high related to ORFV strains previously shown to circulate in Southern Italy.References
Ara M., Zaballos P., Sánchez M., Querol I., Zubiri M.L., Simal E. & Hörndler C. 2008. Giant and recurrent orf virus infection in a renal transplant recipient treated with imiquimod. J Am Acad Dermatol, 58, S39-S40.
Cardeti G., Damiani A., Ponticello L., Cittadini M. & Demetrio A. 2005. Virus Orf da pecore e capre nelle regioni Lazio e Toscana: studio del gene VEGF-E e costruzione di un albero filogenetico. In Atti del Workshop Nazionale di Virologia Veterinaria “Diagnostica ed epidemiologia delle infezioni virali degli animali” (S. Babsa, I. Purificato & F.M. Ruggeri, eds) Istituto Superiore di Sanità, Roma.
Davari S.A., Sayyari M. & Mohammadi A. 2013. Genetic analysis of the viral agents causing muzzle crust in small ruminants of Shiraz, Iran. Bulg J Vet Med, 16, 159-169.
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. 2004. Genomes of the parapoxviruses ORF virus and bovine papular stomatitis virus. J Virol, 78, 168-177.
Doane F.W. & Anderson N. 1987. Methods for preparing specimens for electron microscopy. In Electron microscopy in diagnostic virology. A practical guide and atlas. Cambridge University Press, Cambridge, 14-19.
Friederichs S., Krebs S., Blum H., Wolf E., Lang H., von Buttlar H. & Büttner M. 2014. Comparative and retrospective molecular analysis of Parapoxvirus (PPV) isolates. Virus Res, 181, 11-21.
Gallina L., Scagliarini L., McInnes C.J., Guercio A., Purpari G., Prosperi S. & Scagliarini A. 2008. Parapoxvirus in goats: experimental infection and genomic analysis. Vet Res Commun, 32, S203-S205.
Guindon S. & Gascuel O. 2003. A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Syst Biol, 52, 696-704.
Hosamani M., Scagliarini A., Bhanuprakash V., McInnes C.J. & Singh R.K. 2009. Orf: an update on current research and future perspectives. Expert Rev Anti Infect Ther, 7, 879-893.
Lojkic I., Cac Z., Beck A., Bedekovic T., Cvetnic Z. & Sostaric B. 2010. Phylogenetic analysis of Croatian orf viruses isolated from sheep and goats. Virol J, 7, 314.
Kottaridi C., Nomikou K., Lelli R., Markoulatos P. & Mangana O. 2006a. Laboratory diagnosis of contagious ecthyma: comparison of different PCR protocols with virus isolation in cell culture. J Virol Methods, 134, 119-124.
Kottaridi C., Nomikou K., Teodori L., Savini G., Lelli R., Markoulatos P. & Mangana O. 2006b. Phylogenetic correlation of Greek and Italian orf virus isolated based on VIR gene. Vet Microbiol, 116, 310-316.
Kuhl J.T., Huerter C.J. & Hashish H. 2003. A case of human orf contracted from a deer. Cutis, 71, 288-290.
Mahmoud M., Abdelrahman K. & Soliman H., 2010. Molecular and virological studies on contagious pustular dermatitis isolates from Egyptian sheep and goats. Res Vet Sci, 89, 290-294.
Mazur C. & Machado R.D. 1989. Detection of contagious pustular dermatitis virus of goats in a severe outbreak. Vet Rec, 125, 419-420.
Mazur C. & Machado R.D. 1990. The isolation and identification of the contagious ecthyma virus of caprines in cell cultures. Rev Microbiol Sao Paulo, 21, 127-130.
Mercer A.A., Ueda N., Friederichs S.M., Hofmann K., Fraser K.M., Bateman T. & Fleming S.B. 2006. Comparative analysis of genome sequences of three isolates of Orf virus reveals unexpected sequence variation. Virus Res, 116, 146-158.
Robinson A.J. & Mercer A.A. 1995. Parapoxvirus of red deer: evidence for its inclusion as a new member in the genus parapoxvirus. Virology, 208, 812-815.
Scagliarini A., Vaccari F., Turrini F., Bianchi A., Cordioli P. & Lavazza A. 2011. Parapoxvirus infections of red deer, Italy. Emerg Infect Dis, 17, 684-687.
Tamura K., Stecher G., Peterson D., Filipski A. & Kumar S. 2013. MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Mol Biol Evol, 30, 2725-2729.
Thomas K., Tompkins D.M., Sainsbury A.W., Wood A.R., Dalziel R., Nettleton P.F. & McInnes C.J. 2003. A novel poxvirus lethal to red squirrels (Sciurus vulgaris). J Gen Virol, 84, 3337-3341.
Tryland M., Klein J., Nordøy E.S. & Blix A.S. 2005. Isolation and partial characterization of a Parapoxvirus isolated from a skin lesion of a Weddell seal. Virus Res, 108, 83-87.
Zhang K., Liu Y., Kong H., Shang Y. & Liu X. 2014. Human infection with ORF virus from goats in China, 2012. Vector Borne Zoonotic Dis, 14, 365-367.
Zhao K., Song D., He W., Lu H., Zhang B., Li C., Chen, K. & Gao F. 2010. Identification and phylogenetic analysis of an Orf virus isolated from an outbreak in sheep in the Jilin province of China. Vet Microbiol, 142, 408-415.