Isolation, identification and phylogenetic analysis of a lumpy skin disease virus strain from diseased beef cattle in China

Xinwei Yuan , Xiaowen Xu , Qingni Li , Chen Wang , Zhijie Xiang , Yingyu Chen , Changmin Hu , Aizhen Guo

Animal Diseases ›› 2025, Vol. 5 ›› Issue (1) : 25

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Animal Diseases ›› 2025, Vol. 5 ›› Issue (1) : 25 DOI: 10.1186/s44149-025-00182-x
Original Article

Isolation, identification and phylogenetic analysis of a lumpy skin disease virus strain from diseased beef cattle in China

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Abstract

Lumpy skin disease (LSD) is a highly contagious viral disease in cattle caused by lumpy skin disease virus (LSDV), which belongs to the genus Capripoxvirus (CaPVs) within the family Poxviridae. Since its first outbreak in China in August 2019, LSD has spread widely across mainland China, posing significant threats to the cattle industry. This study aimed to isolate and identify a clinical strain of LSDV via Vero cells. Skin tissue samples from lump lesions were homogenized and inoculated onto cell cultures. After 7 passages, the inoculated cells exhibited typical cytopathic effects (CPEs). PCR amplification of the LSDV132 gene confirmed the presence of LSDV nucleic acid. In addition, quantitative PCR (qPCR) demonstrated a significant increase in viral copy number over time. Transmission electron microscopy (TEM) revealed typical brick-shaped viral particles. Furthermore, an indirect immunofluorescence assay (IFA) of infected Vero cells exhibiting CPEs produced a positive reaction with antiserum from cattle naturally infected with LSDV. Additionally, nucleotide similarity analysis of 123 LSDV strains revealed a high degree of similarity (98.4%-100%) among different geographic lineages. Nucleotide sequencing and recombination analysis of the LSDV011 gene from LSDV/China/HB01/2020 revealed close similarity to Asian strains and revealed a recombination event. Furthermore, similarity plot analysis confirmed two genomic exchange sites at nucleotide positions 120 and 762 within the LSDV011 gene. Recombination events between 65 Asian LSDV strains and 13 goatpox virus (GTPV) strains have raised safety concerns regarding the use of attenuated goatpox vaccines, highlighting the need for novel and safer LSDV vaccines. In summary, this study successfully isolated a clinical LSDV strain, demonstrating its evolutionary status and providing crucial insights for LSD control in the cattle industry.

Keywords

Lumpy skin disease (LSD) / Lumpy skin disease virus (LSDV) / Recombination / Virus isolation / Phylogenetic analysis

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Xinwei Yuan, Xiaowen Xu, Qingni Li, Chen Wang, Zhijie Xiang, Yingyu Chen, Changmin Hu, Aizhen Guo. Isolation, identification and phylogenetic analysis of a lumpy skin disease virus strain from diseased beef cattle in China. Animal Diseases, 2025, 5(1): 25 DOI:10.1186/s44149-025-00182-x

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References

[1]

AbebawB. Prevalence of lumpy skin disease in Africa: A systematic review and meta-analysis from 2007 to 2023. Veterinary Medicine International., 2024, 2024: 9991106

[2]

AbutarbushSM, AbabnehMM, Al ZoubiIG, Al SheyabOM, Al ZoubiMG, AlekishMO, Al GharabatRJ. Lumpy skin disease in Jordan: Disease emergence, clinical signs, complications and preliminary-associated economic losses. Transboundary and Emerging Diseases, 2015, 62(5): 549-554

[3]

AktherM, AkterSH, SarkerS, AleriJW, AnnandaleH, AbrahamS, UddinJM. Global burden of lumpy skin disease, outbreaks, and future challenges. Viruses, 2023, 15(9): 1861

[4]

AnnandaleCH, HolmDE, EbersohnK, VenterEH. Seminal transmission of lumpy skin disease virus in heifers. Transboundary and Emerging Diseases, 2014, 61(5): 443-448

[5]

ArjkumpaO, SuwannaboonM, BoonrawdM, PunyawanI, LaobannuP, YantaphanS, BungwaiA, PonyiumV, SuwankitwatN, BoonpornprasertP, et al.. First emergence of lumpy skin disease in cattle in Thailand, 2021. Transboundary and Emerging Diseases, 2021, 68(6): 3002-3004

[6]

Azeem S, Sharma B, Shabir S, Akbar H, Venter E. 2022. Lumpy skin disease is expanding its geographic range: A challenge for Asian livestock management and food security. Veterinary journal (London, England: 1997) 279: 105785. https://doi.org/10.1016/j.tvjl.2021.105785.

[7]

BadhySC, ChowdhuryMGA, SettypalliTBK, CattoliG, LamienCE, FakirMAU, AkterS, OsmaniMG, TalukdarF, BegumN, KhanIA, RashidMB, SadekuzzamanM. Molecular characterization of lumpy skin disease virus (LSDV) emerged in Bangladesh reveals unique genetic features compared to contemporary field strains. BMC Veterinary Research, 2021, 17(1): 61

[8]

ChihotaCM, RennieLF, KitchingRP, MellorPS. Attempted mechanical transmission of lumpy skin disease virus by biting insects. Medical and Veterinary Entomology, 2003, 17(3): 294-300

[9]

European Food Safety Authority (EFSA); Calistri P, De Clercq K, Gubbins S, Klement E, Stegeman A, Cortiñas Abrahantes J, Marojevic D, Antoniou SE, Broglia A 2020 Lumpy skin disease epidemiological report IV: Data collection and analysis EFSA Journal. European Food Safety Authority 18 2 e06010 https://doi.org/10.2903/j.efsa.2020.6010

[10]

Di Giuseppe A, Zenobio V, Dall'Acqua F, Di Sabatino D, Calistri P. 2024. Lumpy skin disease. The Veterinary clinics of North America. Food animal practice 40(2): 261–276. https://doi.org/10.1016/j.cvfa.2024.01.002.

[11]

HagaIR, ShihBB, ToreG, PoloN, RibecaP, Gombo-OchirD, ShuraG, TserenchimedT, EnkhboldB, PurevtserenD, et al.. Sequencing and Analysis of Lumpy Skin Disease Virus Whole Genomes Reveals a New Viral Subgroup in West and Central Africa. Viruses, 2024, 16(4): 557

[12]

HunterP, WallaceD. Lumpy skin disease in southern Africa: A review of the disease and aspects of control. Journal of the South African Veterinary Association, 2001, 72(2): 68-71

[13]

IssimovA, KutumbetovL, OrynbayevMB, KhairullinB, MyrzakhmetovaB, SultankulovaK, WhitePJ. Mechanical transmission of lumpy skin disease virus by Stomoxys Spp (Stomoxys Calsitrans, Stomoxys Sitiens, Stomoxys Indica), Diptera: Muscidae. Animals, 2020, 10(3): 477

[14]

KlausnerZ, FattalE, KlementE. Using synoptic systems' typical wind trajectories for the analysis of potential atmospheric long-distance dispersal of lumpy skin disease virus. Transboundary and Emerging Diseases, 2017, 64(2): 398-410

[15]

KononovA, ByadovskayaO, KononovaS, YashinR, ZinyakovN, MischenkoV, PerevozchikovaN, SpryginA. Detection of vaccine-like strains of lumpy skin disease virus in outbreaks in Russia in 2017. Archives of Virology, 2019, 164(6): 1575-1585

[16]

KononovA, PrutnikovP, ShumilovaI, KononovaS, NesterovA, ByadovskayaO, PestovaY, DievV, SpryginA. Determination of lumpy skin disease virus in bovine meat and offal products following experimental infection. Transboundary and Emerging Diseases, 2019, 66(3): 1332-1340

[17]

KrotovaA, ByadovskayaO, ShumilovaI, van SchalkwykA, SpryginA. An in-depth bioinformatic analysis of the novel recombinant lumpy skin disease virus strains: From unique patterns to established lineage. BMC Genomics, 2022, 23(1): 396

[18]

KumarN, ChanderY, KumarR, KhandelwalN, RiyeshT, ChaudharyK, ShanmugasundaramK, KumarS, KumarA, GuptaMK, et al.. Isolation and characterization of lumpy skin disease virus from cattle in India. PLoS ONE, 2021, 16(1): e0241022

[19]

LeeSW, MarkhamPF, CoppoMJ, LegioneAR, MarkhamJF, NoormohammadiAH, BrowningGF, FicorilliN, HartleyCA, DevlinJM. Attenuated vaccines can recombine to form virulent field viruses. Science, 2012, 337(6091): 188

[20]

LubingaJC, TuppurainenES, StoltszWH, EbersohnK, CoetzerJA, VenterEH. Detection of lumpy skin disease virus in saliva of ticks fed on lumpy skin disease virus-infected cattle. Experimental & Applied Acarology., 2013, 61(1): 129-138

[21]

Martin DP, Posada D, Crandall KA, Williamson C. 2005. A modified bootscan algorithm for automated identification of recombinant sequences and recombination breakpoints. AIDS Res Hum Retroviruses. 21(1): 98–102. https://doi.org/10.1089/aid.2005.21.98.

[22]

MojeN, SeifuA, HailegebrealG, SheguD, MontagnaroS, FerraraG. Serological and community awareness study of lumpy skin disease in different agro-ecological zones of Sidama regional state. Southern Ethiopia. Animals., 2024, 14(12): 1782

[23]

MoudgilG, ChadhaJ, KhullarL, ChhibberS, HarjaiK. Lumpy skin disease: Insights into current status and geographical expansion of a transboundary viral disease. Microbial Pathogenesis, 2024, 186, 106485

[24]

PosadaD, CrandallKA. Evaluation of methods for detecting recombination from DNA sequences: Computer simulations. Proc Natl Acad Sci U S a., 2001, 98(24): 13757-13762

[25]

Rhazi H, Safini N, Mikou K, Alhyane M, Lenk M, Tadlaoui KO, Elharrak M. 2021. Comparative sensitivity study of primary cells, vero, OA3.Ts and ESH-L cell lines to lumpy skin disease, sheeppox, and goatpox viruses detection and growth. Journal of virological methods 293: 114164. https://doi.org/10.1016/j.jviromet.2021.114164.

[26]

SaegermanC, BertagnoliS, MeyerG, GanièreJP, CaufourP, De ClercqK, JacquietP, FourniéG, HautefeuilleC, EtoreF, CasalJ. Risk of introduction of lumpy skin disease in France by the import of vectors in animal trucks. PLoS ONE, 2018, 13(6): e0198506

[27]

ShumilovaI, NesterovA, ByadovskayaO, PrutnikovP, WallaceDB, MokeevaM, ProninV, KononovA, ChvalaI, SpryginA. A recombinant vaccine-like strain of lumpy skin disease virus causes low-level infection of cattle through virus-inoculated feed. Pathogens, 2022, 11(8): 920

[28]

SmarakiN, JogiHR, KamothiDJ, SavsaniHH. An insight into emergence of lumpy skin disease virus: A threat to Indian cattle. Archives of Microbiology, 2024, 206(5): 210

[29]

SongY, ZuoO, ZhangG, HuJ, TianZ, GuanG, LuoJ, YinH, ShangY, DuJ. Emergence of lumpy skin disease virus infection in yaks, cattle-yaks, and cattle on the Qinghai-Xizang plateau of China. Transboundary and Emerging Diseases, 2024, 2024: 2383886

[30]

SpryginA, ArtyuchovaE, BabinY, PrutnikovP, KostrovaE, ByadovskayaO, KononovA. Epidemiological characterization of lumpy skin disease outbreaks in Russia in 2016. Transboundary and Emerging Diseases, 2018, 65(6): 1514-1521

[31]

SpryginA, BabinY, PestovaY, KononovaS, WallaceDB, Van SchalkwykA, ByadovskayaO, DievV, LozovoyD, KononovA. Analysis and insights into recombination signals in lumpy skin disease virus recovered in the field. PLoS ONE, 2018, 13(12): e0207480

[32]

SpryginA, PestovaY, BjadovskayaO, PrutnikovP, ZinyakovN, KononovaS, RuchnovaO, LozovoyD, ChvalaI, KononovA. Evidence of recombination of vaccine strains of lumpy skin disease virus with field strains, causing disease. PLoS ONE, 2020, 15(5): e0232584

[33]

Sprygin A, Pestova Y, Prutnikov P, Kononov A. 2018. Detection of vaccine-like lumpy skin disease virus in cattle and Musca domestica L. flies in an outbreak of lumpy skin disease in Russia in 2017. Transboundary and emerging diseases 65(5): 1137–1144. https://doi.org/10.1111/tbed.12897.

[34]

SudhakarSB, MishraN, KalaiyarasuS, JhadeSK, SinghVP. Genetic and phylogenetic analysis of lumpy skin disease viruses (LSDV) isolated from the first and subsequent field outbreaks in India during 2019 reveals close proximity with unique signatures of historical Kenyan NI-2490/Kenya/KSGP-like field strains. Transboundary and Emerging Diseases, 2022, 69(4): e451-e462

[35]

TranHTT, TruongAD, DangAK, LyDV, NguyenCT, ChuNT, HoangTV, NguyenHT, NguyenVT, DangHV. Lumpy skin disease outbreaks in Vietnam, 2020. Transboundary and Emerging Diseases, 2021, 68(3): 977-980

[36]

VandenbusscheF, MathijsE, PhilipsW, SaduakassovaM, De LeeuwI, SultanovA, HaegemanA, De ClercqK. Recombinant LSDV strains in Asia: Vaccine spillover or natural emergence?. Viruses, 2022, 14(7): 1429

[37]

WangJ, XuZ, WangZ, LiQ, LiangX, YeS, ChengK, XuL, MaoJ, WangZ, MengW, SunY, JiaK, LiS. Isolation, identification and phylogenetic analysis of lumpy skin disease virus strain of outbreak in Guangdong. China. Transboundary and Emerging Diseases, 2022, 69(5): e2291-e2301

[38]

WeiYR, MaWG, WangP, WangW, SuXH, YangXY, MiXY, WuJY, HuangJ. Retrospective genomic analysis of the first lumpy skin disease virus outbreak in China (2019). Frontiers in Veterinary Science, 2023, 9: 1073648

[39]

WhittleL, ChapmanR, WilliamsonAL. Lumpy skin disease-an emerging cattle disease in Europe and Asia. Vaccines, 2023, 11(3): 578

[40]

WOAH, Manual of diagnostic tests and vaccines for terrestrial animals 2023-lumpy skin diseases, WOAH Terrestrial Manual, Paris, 2023.

[41]

Xu X, Zhao W, Xiang Z, Wang C, Qi M, Zhang S, Geng Y, Zhao Y, Yang K, Zhang Y, Guo A, Chen Y. 2024. Prevalence, molecular characteristics and virulence identification of bovine parainfluenza virus type 3 in China. Viruses 16(3): 402. https://doi.org/10.3390/v16030402.

[42]

YuanXW, ZhaoH, JiWF, YanXH, XiangZJ, YangL, GengYC, ChenYY, ChenJG, ChenX, ZhangL, HuCM, GuoAZ. Preparation of a monoclonal antibody against recombinant LSDV034 protein and its application in detecting lumpy skin disease virus through a competitive enzyme-linked immunosorbent assay (cELISA). Animal Diseases, 2024, 4(1): 23

[43]

Yuan XW, Wang Y, Adili Abulaiti, Li JK, Chen YY, Guo AZ. 2023. Research progress on prevalence, prevention and control of lumpy skin disease in cattle. Journal of Huazhong Agricultural University 42 (02): 9–16. https://doi.org/10.13300/j.cnki.hnlkxb. 2023. 02. 002.

[44]

Zaghloul M, Azooz M, Ali S, Soliman H, Sayed M, Kafafy M, Morsy A. 2022. Sequencing and phylogenetic analysis of GPCR, RPO30, P32 and EEV glycoprotein genes of lumpy skin disease virus recent isolates in Egypt. Journal of Virological Sciences1: 1–11. https://doi.org/10.17582/journal.jvs/sp1.1.11.

[45]

ZhangMM, SunYJ, LiuWX, LiuRQ, WangXJ, BuZG. Isolation and identification of lumpy skin disease virus from the first outbreak in China. Chinese Journal of Preventive Veterinary Medicine, 2020, 42(10): 1058-1061

[46]

Zhou ZY, Du JG, Xin RL, Zhang JW, Pan CF, Yin CS, Chen XY, Zhu Z. 2024. Isolation and identification of three lumpy skin disease viruses in China and their GPCR gene analysis. Acta Veterinaria et Zootechnica Sinica 1–19. https://doi.org/10.11843/j.issn.0366-6964.2024.12.025.

Funding

National Key Research and Development Program of China(2023YFD1802505)

Chinese Agricultural Research System of MOF and MARA(#CARS-37)

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