Isolation and characterization of lumpy skin disease virus from cattle in India

Kumar, N.; Chander, Y.; Kumar, R.; Khandelwal, N.; Riyesh, T.; Chaudhary, K.; Shanmugasundaram, K.; Kumar, S.; Kumar, A.; Gupta, M. K.; Pal, Y.; Barua, S.; Tripathi, B. N.

doi:10.1101/2020.10.08.331124

bioRxiv · 10.1101/2020.10.08.331124

Isolation and characterization of lumpy skin disease virus from cattle in India

Kumar, N.·Chander, Y.·Kumar, R.·Khandelwal, N.·Riyesh, T.·Chaudhary, K.·Shanmugasundaram, K.·Kumar, S.·Kumar, A.·Gupta, M. K.·Pal, Y.·Barua, S.·Tripathi, B. N.

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Abstract

Lumpy skin disease (LSD) has devastating economic impact. During the last decade, LSD had spread to climatically new and previously disease-free countries, which also includes its recent emergence in the Indian subcontinent (2019). This study deals with the LSD outbreak(s) from cattle in Ranchi (India). Virus was isolated from the scabs (skin lesions) in the primary goat kidney cells. Phylogenetic analysis based on nucleotide sequencing of LSD virus (LSDV) ORF011, ORF012 and ORF036 suggested that the isolated virus (LSDV/Bos taurus-tc/India/2019/Ranchi) is closely related to Kenyan LSDV strains. Further, we adapted the isolated virus in Vero cells, an alternative cell line that was found suitable for LSDV propagation. In addition, kinetics of viral DNA synthesis and one-step growth curve analysis suggested that LSDV initiates synthesizing its genome at [~]24 hours post-infection (hpi) with a peak level at [~]96 hpi whereas evidence of progeny virus particles was observed at 36-48 hours (h) with a peak titre at [~]120 h. This study reports the first successful isolation of LSDV in India, besides describing alternative cells for virus isolation (primary goat kidney cells) and in vitro propagation (Vero cells) as well as providing some insights on LSDV life cycle. Author SummaryThis study describes the first successful isolation of LSDV in India. Nucleotide sequencing of LSDV ORF011, ORF012 and ORF036 suggested that the isolated virus is closely related to Kenyan LSDV strains. This is in accordance with the previous (only) study, thereby suggesting that a single LSDV strain is circulating in India. For the first time, we employed primary goat kidney (PGK) cells for the isolation of LSDV, where PGK cells were found more sensitive for LSDV isolation as compared to the primary lamb testicle (PLT) and primary goat testicle (PGT) cells. We also adapted the isolated virus in Vero cells and demonstrated that Vero cells are also suitable for in vitro propagation of LSDV. Besides, we also provide some insights on the LSDV life cycle.

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Kumar, N., Chander, Y., Kumar, R., Khandelwal, N., Riyesh, T., Chaudhary, K., Shanmugasundaram, K., Kumar, S., Kumar, A., Gupta, M. K., Pal, Y., Barua, S., Tripathi, B. N.. 2020-10-08. Isolation and characterization of lumpy skin disease virus from cattle in India. https://doi.org/10.1101/2020.10.08.331124

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