The aim of this work was to evaluate the immunogenicity and neutralizing activity of sheep pox virus (SPPV; genus Capripoxvirus, family Poxviridae) structural proteins as candidate subunit vaccines to control sheep pox disease. SPPV structural proteins were identified by sequence homology with proteins of vaccinia virus (VACV) strain Copenhagen. Four SPPV proteins (SPPV-ORF 060, SPPV-ORF 095, SPPV-ORF 117, and SPPV-ORF 122), orthologs of immunodominant L1, A4, A27, and A33 VACV proteins, respectively, were produced in Escherichia coli. Western blot analysis revealed the antigenic and immunogenic properties of SPPV-060, SPPV-095, SPPV-117 and SPPV-122 proteins when injected with adjuvant into experimental rabbits. Virus-neutralizing activity against SPPV in lamb kidney cell culture was detected for polyclonal antisera raised to SPPV-060, SPPV-117, and SPPV-122 proteins. To our knowledge, this is the first report demonstrating the virus-neutralizing activities of antisera raised to SPPV-060, SPPV-117, and SPPV-122 proteins.

Objective: A chloroplast transgenic approach was assessed in order to produce a structural protein SPPV117 of sheep pox virus in Nicotiana tabacum for the future development of a plant-based subunit vaccine against sheep pox. Results: Two DNA constructs containing SPPV117 coding sequence under the control of chloroplast promoter and terminator of psbA gene or rrn promoter and rbcL terminator were designed and inserted into the chloroplast genome by a biolistic method. The transgenic plants were selected via PCR analysis. Northern and Western blot analysis showed expression of the transgene at transcriptional and translational levels, respectively. The recombinant protein accumulated to about 0.3% and 0.9% of total soluble protein in leaves when expressed from psbA and rrn promoter, respectively. Plant-produced SPPV117 protein was purified using metal affinity chromatography and the protein yield was 19.67 ± 1.25 µg g⁻¹ (FW)_. The serum of a sheep infected with the virus recognised the chloroplast-produced protein indicating that the protein retains its antigenic properties. Conclusions: These results demonstrate that chloroplasts are a suitable system for the production of a candidate subunit vaccine against sheep pox.

Viroids represent obligate plant pathogens composed exclusively of non-protein coding small single-stranded RNAs that cause high economic losses worldwide. A field survey was carried out to assess the incidence of the peach latent mosaic viroid (PLMVd) in southeastern Kazakhstan, the region of the country where fruit trees are mainly grown. Of 246 stone fruit trees, 20 (8.13%) were infected with the PLMVd. The incidence of the PLMVd in the peach (19.23%; 15/78) was significantly higher than that in the apricot (6.76%; 5/74; p = 0.0234). Eight of the detected viroids were cloned and used for full-genome sequencing. The nucleotide sequence similarity of the selected isolates found in Kazakhstan was 83.9–100%. A phylogenetic analysis indicated three clusters for the Kazakhstani isolates of the PLMVd. Three groups of Kazakhstani viroids differed in their predicted secondary structure. During the survey, the PLMVd was detected and genetically characterized for the first time in Kazakhstan. The obtained results indicate the need to develop state control measures for the PLMVd, including regular monitoring surveys. We identified several SNPs of the PLMVd that had not been previously described. The results may be useful in optimizing diagnostic approaches for detecting stone fruit viroids and preventing their spread through propagation material.