Molecular phylogeny of Capripoxviruses based on major immunodominant protein (P32) reveals circulation of host specific sheeppox and goatpox viruses in small ruminants of India.

Sheeppox and goatpox are highly contagious viral diseases of small ruminants causing severe economic losses to the livestock farmers. The disease is enzootic in Asia including India, Middle East and African countries. In the present study, a total of 28 isolates from twenty five sheeppox and goatpox disease outbreaks were phylogenetically analyzed based on P32 gene/protein along with homology modeling and docking using heparan sulfate and UDP-glucose. Three distinct lineage-specific clusters as per their host origin were recorded. Multiple sequence analysis of P32 gene revealed that genetically similar sheeppox virus (SPPV) and goatpox virus (GTPV) strains are circulating in India. Phylogenetically, Lumpy skin disease (LSDV) and SPPV had a closer genetic relationship than GTPV. Comparative sequence alignment indicated conservation of various motifs such as glycosaminoglycan (GAG), chemokine like motif (CX3C) and Asp-Glu-any other residue-Asp (D/ExD), as well as viral specific signature residues in SPPV and GTPV isolates. Structurally, P32 protein of SPPV and GTPV with mixed α helices and ß sheets resembled with crystal structure of homologue vaccinia virus H3L protein. Docking studies in P32 protein of SPPV and GTPV revealed conserved binding pattern with heparan sulfate which is involved in the virus attachment and varied glycosyltransferase fold with UDP-glucose. These findings may help in development of suitable vaccines/diagnostics and therapeutics against capripoxviruses.

Molecular characterization of sheeppox virus from outbreaks in Karnataka, India.

AIM: This study aimed to characterize sheeppox virus (SPPV) using the P32 gene of the Capripoxvirus (CaPVs). MATERIALS AND METHODS: Clinical samples of skin, scabs, and nasal swab from suspected outbreaks Horalagallu (n=13) and Gerahalli (n=11) at Ramanagara district in Karnataka were collected. All the samples were initially subjected to genus-specific diagnostic polymerase chain reaction (PCR). The pooled clinical samples from each outbreak were also subjected to virus isolation. The isolates were confirmed by CaPVs genotyping PCR targeting the full-length P32 gene, followed by sequencing and phylogenetic analysis. RESULTS: The clinical signs and lesions varied from mild to severe degree with no specificity between age and sex. Specific cytopathic changes in cell morphology were observed in infected Vero cells from both outbreaks, which were confirmed by PCR. The complete P32 gene from two outbreaks was successfully amplified with the expected amplicon size of 1006bp. The sequencing and phylogenetic analysis revealed that both the outbreaks were due to SPPV and shared high similarity with published SPPVs from Karnataka and other parts of India. CONCLUSION: The current study showed that complete P32 gene-based genotypic PCR assay can be used for genetic characterization and molecular epidemiology of both sheeppox and goatpox diseases and also to differentiate the causative agents. The sequence analysis revealed 100% similarity among the two outbreak isolates suggesting the same strain of the virus and common source of infection for the outbreaks.