Molecular analysis of parapoxvirus envelope genes was performed. BPSV V660, Iwate/bovine/2007, 7 of the amplicons identified in this study (ACF) and BPSV 9108 (Fig. 2). The I site, because of the T61C nucleotide substitution. This substitution was also present in BPSV 9108. In the deduced amino acid sequences for F and G, the presence of an I301V substitution has the potential to confer a change in antigenicity. Generally, the virus envelope antigen is highly variable, because of its exposure to host immunological pressures. These substitutions might be the result of exposure to some as yet unidentified immunological pressures. The nucleotide sequence of amplicon B was identical to that of BPSV Iwate/bovine/2007 detected in a calf from Iwate Prefecture in 2007 . Based on the phylogenetic tree we constructed, seven of the detected amplicons (ACG) could be considered BPSV; however, the sequence of amplicon H was most similar to that of PCPV (Fig. 3). Within the larger cluster, F and G formed a subgroup along with BPSV 9108. The nucleotide and deduced amino acid sequence identities for the amplicons and parapoxvirus reference sequences are shown in Table 1. Fig. 2. Nucleotide sequence alignment of partial regions of the envelope gene. Nucleotides identical to BPSV are represented by dots. ACH, sequenced amplicons; BPSV, bovine papular stomatitis virus strain V660; Iwate/bovine/2007, BPSV strain Iwate/bovine/2007; 9108, … Fig. 3. Phylogenetic tree derived from the deduced amino acid sequences of partial envelope gene regions. ACH, sequenced amplicons; V660, BPSV strain V660; VR634, pseudocowpox virus strain VR634; DPV, parapoxvirus of red deer in New Zealand strain DPV; … Three swabs from three calves (farms 1, 2 and 6) showed CPE in BT cell cultures after a second blind passage. Other swabs showed no signs of CPE. The molecular characteristics of three isolates were identical to buy NU6027 those of amplicons (A, B and F) from the swab of the same calves. Parapoxvirus envelope genes were detected in eight calves from eight farms. Six of these calves were symptomatic, and two were asymptomatic. Because the asymptomatic calves had been raised with one symptomatic calf at each farm, the calves might have been subclinically infected with buy NU6027 the virus. Parapoxvirus infections with clinical symptoms in cattle have rarely been reported in this prefecture , although a high rate of antibodies positive against parapoxvirus has been found . The increase in the level of detection is buy NU6027 likely a consequence of increased surveillance by veterinarians and farmers following a foot-and-mouth disease outbreak in the spring of 2010 in Japan. This would suggest that parapoxvirus infections are widespread among cattle in Iwate. Infections occur irrespective of age with relapses common, because infections confer no significant immunity . However, clinical symptoms were only observed in calves from 2- to 10-month-old. This finding is in accordance with a previous report that found that infections were more common in calves than adult cattle . Five (ACE) of the eight samples examined in our study were classified as BPSV, based on amino acid identities with the corresponding regions in BPSV V660 . In particular, amplicon B showed 100% nucleotide identity to the amplicon from Iwate/bovine/2007, in spite of being isolated in different years. This strongly suggests that this examined region encoding an envelope protein is highly conserved. By contrast, the presence of C19orf40 BPSV, which can be digested with I and 49: 4397C4400. doi: 10.1128/JCM.05281-11 [PMC free article] [PubMed] [Cross Ref] 3. Fraser C. M., Savan M. 1962. Bovine papular stomatitis, a note on its diagnosis and experimental transmission in Ontario. 3: 107C111 [PMC free article] [PubMed] 4. Friedman-Kien A. E., Rowe W. P., Banfield W. G. 1963. Milkers nodules: isolation of a poxvirus from a human case. 140: 1335C1336. doi: 10.1126/science.140.3573.1335 [PubMed] [Cross Ref] 5. Ginn P. E., Mansell J. E. K. L., Rakich P. M. 2007. Parapoxviral diseases. pp. 665C668. 137: 404C410 [PubMed] 7. Inoshima Y., Morooka A.,.