Spatial and Temporal Genetic Homogeneity of Orf Viruses Infecting Japanese Serows (Capricornis crispus)

Abstract

ABSTRACT. This study genetically characterized orf viruses (ORFVs) isolated from recent outbreaks in Japanese serows (Capricornis crispus) in 2007 and 2008, and from earlier outbreaks from 1985 to 2001. Nucleotide sequences of genes for the viral envelope, vascular endothelial growth factor (VEGF), and virus interferon resistance (VIR) were determined in two ORFVs isolated from recent outbreaks and in eight from earlier outbreaks. No deletions or insertions were observed in these genes. Surprisingly, the amino acid sequences of the envelope and VIR genes were identical among the 10 ORFVs, and only one amino acid substitution in the VEGF gene was found in one of the two recent ORFVs (2007). Genotyping by differential PCR for the A32L gene, which encodes an ATPase, classified all of the 10 ORFVs into the same genotype. In an ORFV isolated from a sheep in 1970, the three sequenced genes were almost the same as in the ORFVs isolated from Japanese serows, and the ORFV in 1970 was classified into the same genotype as the ORFVs from Japanese serows. These results suggest that recent outbreaks in Japanese serows are caused by ORFVs genetically closely related to the viruses in earlier outbreaks and that these genetically stable ORFVs have circulated among Japanese serows over a long period of time.

On-site Visual Diagnosis of Parapoxvirus Infection Using a Portable Cordless Incubator

“…Six parapoxvirus strains isolated from sheep [10][11][12] and wild Japanese serows (Capricornis crispus) [13][14][15] were used in this study (Table 1). All parapoxviruses were propagated in primary fetal bovine muscle (FBM) cells or fetal lamb lung (FLL) cells.…”

“…2A). Since the viruses used for the assay were isolated from sheep and wild Japanese serows from different regions and countries between the 1970s to the 2000s (Table 1), [10][11][12][13][14][15] it is considered that the incubator developed in this study, as well as the LAMP protocol described above, could be used to diagnose orf virus infection in sheep, goats, Japanese serows, and humans around the world. In a comparison of the three color reagents used as visual markers in reactions performed on the developed incubator and a Loopamp EXIA real-time turbidimeter, detectability by the naked eye was similar and determining whether the sample tested positive or negative was straightforward ( Fig.…”

Establishment of an on-site diagnostic procedure for detection of orf virus from oral lesions of Japanese serows (<i>Capricornis crispus</i>) by loop-mediated isothermal amplification

“…For comparison of quality and applicability to DNA amplification, DNA was also extracted from the same samples by a PureLink Genomic DNA Mini Kit (PureLink Kit, Invitrogen, Carlsbad, CA, U.S.A.), a common laboratory kit, according to the manufacturer’s instructions. Moreover, six orf virus strains isolated from sheep, NZ2 [28], Iwate [18] and HIS [14], and from wild Japanese serows, S-1 [30], IJS081 [12] and GE [7], were also added to goat saliva, and DNA was extracted by a PURE Kit from each. As a negative control, DNA was also extracted from goat saliva without adding virus.…”

“…Diagnosis of orf virus infection in Japanese serows as well as domestic animals and humans is currently based on laboratory examinations, such as electron microscopic observation of viral particles [26], immunohistochemical detection of viral antigen [25, 27], virus isolation [7, 9, 12, 30], detection of viral DNA by polymerase chain reaction (PCR) [8], and serological tests by agar gel immunodiffusion test and enzyme-linked immunosorbent assay [10, 11]. Since free-ranging Japanese serows mainly live in mountainous areas, samples for diagnosis must be moved from these areas to laboratories in towns, and it is a time-consuming process.…”

Multifocal Cutaneous Orf Virus Infection in Goats in the Amazon Region, Brazil

“…The virus was detected using PCR amplification of the whole B2L open reading frame as previously described (Hosamani et al 2006), using 2 lL of the supernatants as a template. In addition, to improve our analysis, the virus interferon resistance gene (VIR) and the vascular endothelial growth factor gene (VEGF) were also amplified, as described previously (Inoshima et al 2010). A Brazilian sheep ORFV scab, MT05 (Abrahã o et al 2009), was used as PCR positive control.…”