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

Naveen Kumar,Sanjay Barua,Khushboo Chaudhary,Karuppusamy Shanmugasundaram,Anand Kumar,Nitin Khandelwal,Yogesh Chander,Thachamvally Riyesh,Bhupendra N Tripathi,Madhurendu K Gupta,Ram Kumar,Sanjit Kumar,Yash Pal,Pierre Roques

doi:10.1371/journal.pone.0241022

Naveen Kumar, Sanjay Barua + Show 12 more

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https://doi.org/10.1371/journal.pone.0241022

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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. Infection of the isolated LSDV to Vero cells did not produce cytopathic effect (CPE) until the 4th blind passage, but upon adaptation, it produced high viral titres in the cultured cells. The kinetics of viral DNA synthesis and one-step growth curve analysis suggested that Vero cell-adapted 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. To the best of our knowledge, this study describes the first successful isolation of LSDV in India, besides providing insights into the life cycle Vero cell-adapted LSDV.

Highlights

Lumpy skin disease (LSD) is a trans-boundary animal viral disease which causes considerable financial losses to the livestock industry
Primary goat testicle (PGT) cells [16], primary goat kidney (PGK) cells [17], primary lamb testicle (PLT) cells [16] and Madin-Darby bovine kidney (MDBK) cells [18] were available at National Centre for Veterinary Type Cultures (NCVTC), Hisar and grown in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with antibiotics and 10% foetal calf serum
Lumpy skin disease was restricted to Africa wherein it led to several devastating pandemics in several countries, thereby threatening food security and increasing poverty [22, 23]

Summary

Introduction

Lumpy skin disease (LSD) is a trans-boundary animal viral disease which causes considerable financial losses to the livestock industry. It was observed for the first time in Zambia in 1929 [1] and spread rapidly in the cattle population across African countries [reviewed in reference [2]]. The first confirmed transcontinental spread of LSD from the African to Middle-East Asian countries occurred when the disease was reported in Israel in 1989 [3]. By 2015–16, the disease was reported in South-East Europe, the Balkans and the Caucasus [4]. Of late the disease was reported for the first time from India in November 2019 [5]

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