Isolation and molecular characterization of lumpy skin disease virus from hard ticks, Rhipicephalus (Boophilus) annulatus in Egypt.

BACKGROUND: Lumpy skin disease (LSD), a disease of cattle and buffaloes, has recently become widely prevalent in Egypt. The aim of this study was to ascertain the potential role of Rhipicephalus (Boophilus) annulatus ticks in the transmission of this disease. Samples collected from suspected lumpy skin disease virus (LSDV) infected cows that had previously been vaccinated with the Romanian sheep pox virus (SPPV) in various Egyptian governorates were obtained between May to November over two consecutive years, namely 2018 and 2019. Ticks were morphologically identified and the partial cytochrome oxidase subunit I gene (COI) were sequenced, revealing that they were closely related to R. (Boophilus) annulatus. The G-protein-coupled chemokine receptor (GPCR) gene of the LSDV was used to test hard ticks. RESULTS: Two positive samples from Kafr El-Sheikh province and one positive sample from Al-Behera province were reported. BLAST analysis revealed that the positive samples were closely related to the Kazakhstani Kubash/KAZ/16 strain (accession number MN642592). Phylogenetic analysis revealed that the GPCR gene of the LSDV recently circulating in Egypt belongs to a global cluster of field LSDV with a nucleotide identity of 98-100%. LSDV isolation was successfully performed four days after inoculation using 9 to 11-day-old embryonated chicken eggs showing characteristic focal white pock lesions dispersed on the choriallantoic membrane after three blind passages. Intracytoplasmic inclusion bodies, cell rupture, vacuoles in cells, and virus particles ovoid in shape were demonstrated by electron microscopy. CONCLUSION: In this study the role of hard ticks in the transmission of the LSDV to susceptible animals in Egypt was revealed and confirmed by various methods.