1887

Abstract

Since the large-scale outbreak of porcine epidemic diarrhoea (PED) in 2010, caused by the genotype 2 (G2) variant of the porcine epidemic diarrhoea virus (PEDV), pig farms in China, even those vaccinated with the G2b vaccine, have experienced infections from the G2a variant, leading to significant economic losses. This study successfully isolated the G2a strain DY2020 from positive small intestine contents (SICs) by blind passage on Vero cells for four generations. The SICs were taken from Daye, Hubei Province, China. The biological characteristics were identified by indirect immunofluorescence assay (IFA) and transmission electron microscopy (TEM). The growth kinetics of the strain on Vero cells were detected by TCID, and the virus titre could reach 10 TCID ml (SD: 5.07×10). The pathogenicity towards colostrum-deprived piglets was conducted by assessing faecal viral shedding, morphometric analysis of intestinal lesions, and immunohistochemical staining. The results showed that DY2020 was highly virulent to colostrum-deprived piglets, with severe watery diarrhoea and other clinical symptoms appeared at 6 h post-infection (h p.i.), and all died within 30 h. Pathological tissue examination results showed that the lesions mainly occurred in the intestines of piglets, causing pathological changes such as shortening of intestinal villi. In summary, the discovery of the G2a strain DY2020 in this study is of great significance for understanding Hubei PEDV and provides an important theoretical basis for the development of new efficient PEDV vaccines.

Funding
This study was supported by the:
  • Innovative Research Group Project of the National Natural Science Foundation of China (Award 32302866)
    • Principle Award Recipient: ChangLi
Loading

Article metrics loading...

/content/journal/jgv/10.1099/jgv.0.001953
2024-01-25
2024-05-15
Loading full text...

Full text loading...

References

  1. Zhang Y, Chen Y, Zhou J, Wang X, Ma L et al. Porcine epidemic diarrhea virus: an updated overview of virus epidemiology, virulence variation patterns and virus-host interactions. Viruses 2022; 14:2434 [View Article] [PubMed]
    [Google Scholar]
  2. Jung K, Annamalai T, Lu Z, Saif LJ. Comparative pathogenesis of US porcine epidemic diarrhea virus (PEDV) strain PC21A in conventional 9-day-old nursing piglets vs. 26-day-old weaned pigs. Vet Microbiol 2015; 178:31–40 [View Article] [PubMed]
    [Google Scholar]
  3. Pensaert MB, de Bouck P. A new coronavirus-like particle associated with diarrhea in swine. Arch Virol 1978; 58:243–247 [View Article] [PubMed]
    [Google Scholar]
  4. Zhang H, Zou C, Peng O, Ashraf U, Xu Q et al. Global dynamics of porcine enteric coronavirus PEDV epidemiology, evolution, and transmission. Mol Biol Evol 2023; 40:msad052 [View Article] [PubMed]
    [Google Scholar]
  5. Wang D, Fang L, Xiao S. Porcine epidemic diarrhea in China. Virus Res 2016; 226:7–13 [View Article] [PubMed]
    [Google Scholar]
  6. Li M, Pan Y, Xi Y, Wang M, Zeng Q. Insights and progress on epidemic characteristics, genotyping, and preventive measures of PEDV in China: a review. Microb Pathog 2023; 181:106185 [View Article] [PubMed]
    [Google Scholar]
  7. Chen J, Wang C, Shi H, Qiu H-J, Liu S et al. Complete genome sequence of a Chinese virulent porcine epidemic diarrhea virus strain. J Virol 2011; 85:11538–11539 [View Article] [PubMed]
    [Google Scholar]
  8. Chen J, Wang C, Shi H, Qiu H, Liu S et al. Molecular epidemiology of porcine epidemic diarrhea virus in China. Arch Virol 2010; 155:1471–1476 [View Article] [PubMed]
    [Google Scholar]
  9. Hanke D, Pohlmann A, Sauter-Louis C, Höper D, Stadler J et al. Porcine epidemic diarrhea in Europe: in-detail analyses of disease dynamics and molecular epidemiology. Viruses 2017; 9:177 [View Article] [PubMed]
    [Google Scholar]
  10. Vlasova AN, Marthaler D, Wang Q, Culhane MR, Rossow KD et al. Distinct characteristics and complex evolution of PEDV strains, North America, may 2013-February 2014. Emerg Infect Dis 2014; 20:1620
    [Google Scholar]
  11. Liu X, Zhang L, Zhang Q, Zhou P, Fang Y et al. Evaluation and comparison of immunogenicity and cross-protective efficacy of two inactivated cell culture-derived GIIa- and GIIb-genotype porcine epidemic diarrhea virus vaccines in suckling piglets. Vet Microbiol 2019; 230:278–282 [View Article] [PubMed]
    [Google Scholar]
  12. Wang Q, Vlasova AN, Kenney SP, Saif LJ. Emerging and re-emerging coronaviruses in pigs. Curr Opin Virol 2019; 34:39–49 [View Article] [PubMed]
    [Google Scholar]
  13. Yu K, Liu X, Lu Y, Long M, Bai J et al. Biological characteristics and pathogenicity analysis of a low virulence G2A porcine epidemic diarrhea virus. Microbiol Spectr 2023; 11:e0453522 [View Article] [PubMed]
    [Google Scholar]
  14. Zhang L, Yu R, Zhang Z, Zhou P, Lv J et al. Differences in the pathogenicity of Chinese virulent genotype GIIa and GIIb porcine epidemic diarrhea virus strains and the humoral immune status of one- and two-month-old weaned pigs infected with these strains. Arch Virol 2023; 168:97 [View Article] [PubMed]
    [Google Scholar]
  15. Wang S, Yang K, Li C, Liu W, Gao T et al. 4-Phenyl-butyric acid inhibits Japanese encephalitis virus replication via inhibiting endoplasmic reticulum stress response. Viruses 2023; 15:534 [View Article] [PubMed]
    [Google Scholar]
  16. Mei X, Guo J, Fang P, Ma J, Li M et al. The characterization and pathogenicity of a recombinant porcine epidemic diarrhea virus variant ECQ1. Viruses 2023; 15:1492 [View Article] [PubMed]
    [Google Scholar]
  17. Li X, Li Y, Huang J, Yao Y, Zhao W et al. Isolation and oral immunogenicity assessment of porcine epidemic diarrhea virus NH-TA2020 strain: One of the predominant strains circulating in China from 2017 to 2021. Virol Sin 2022; 37:646–655 [View Article] [PubMed]
    [Google Scholar]
  18. Gong L, Li J, Zhou Q, Lin Y, Qin J et al. Insights into recombination-like events leading to outbreaks in USA through a retrospective study of porcine epidemic diarrhea virus isolates from China. Infect Genet Evol 2018; 63:216–218 [View Article] [PubMed]
    [Google Scholar]
  19. Li C, Lu H, Geng C, Yang K, Liu W et al. Epidemic and evolutionary characteristics of swine enteric viruses in South-Central China from 2018 to 2021. Viruses 2022; 14:1420 [View Article] [PubMed]
    [Google Scholar]
  20. Shen Y, Yang Y, Zhao J, Geng N, Liu K et al. Molecular epidemiological survey of porcine epidemic diarrhea in some areas of shandong and genetic evolutionary analysis of S gene. Front Vet Sci 2022; 9:1015717 [View Article] [PubMed]
    [Google Scholar]
  21. Niu T-M, Yu L-J, Zhao J-H, Zhang R-R, Ata EB et al. Characterization and pathogenicity of the porcine epidemic diarrhea virus isolated in China. Microb Pathog 2023; 174:105924 [View Article] [PubMed]
    [Google Scholar]
  22. Liang W, Zhou D, Geng C, Yang K, Duan Z et al. Isolation and evolutionary analyses of porcine epidemic diarrhea virus in Asia. PeerJ 2020; 8:e10114 [View Article] [PubMed]
    [Google Scholar]
  23. Chen J-F, Sun D-B, Wang C-B, Shi H-Y, Cui X-C et al. Molecular characterization and phylogenetic analysis of membrane protein genes of porcine epidemic diarrhea virus isolates in China. Virus Genes 2008; 36:355–364 [View Article] [PubMed]
    [Google Scholar]
  24. Oka T, Saif LJ, Marthaler D, Esseili MA, Meulia T et al. Cell culture isolation and sequence analysis of genetically diverse US porcine epidemic diarrhea virus strains including a novel strain with a large deletion in the spike gene. Vet Microbiol 2014; 173:258–269 [View Article] [PubMed]
    [Google Scholar]
  25. Chen Q, Li G, Stasko J, Thomas JT, Stensland WR et al. Isolation and characterization of porcine epidemic diarrhea viruses associated with the 2013 disease outbreak among swine in the United States. J Clin Microbiol 2014; 52:234–243 [View Article] [PubMed]
    [Google Scholar]
  26. Qian S, Zhang W, Jia X, Sun Z, Zhang Y et al. Isolation and identification of porcine epidemic diarrhea virus and its effect on host natural immune response. Front Microbiol 2019; 10:2272 [View Article] [PubMed]
    [Google Scholar]
  27. Yao X, Qiao W-T, Zhang Y-Q, Lu W-H, Wang Z-W et al. A new PEDV strain CH/HLJJS/2022 can challenge current detection methods and vaccines. Virol J 2023; 20:13 [View Article] [PubMed]
    [Google Scholar]
  28. Li W, van Kuppeveld FJM, He Q, Rottier PJM, Bosch B-J. Cellular entry of the porcine epidemic diarrhea virus. Virus Res 2016; 226:117–127 [View Article] [PubMed]
    [Google Scholar]
  29. Li M, Wu Y, Chen J, Shi H, Ji Z et al. Innate immune evasion of porcine epidemic diarrhea virus through degradation of the FBXW7 protein via the ubiquitin-proteasome pathway. J Virol 2022; 96:e0088921 [View Article] [PubMed]
    [Google Scholar]
  30. Liu X, Zhang Q, Zhang L, Zhou P, Yang J et al. A newly isolated Chinese virulent genotype GIIb porcine epidemic diarrhea virus strain: biological characteristics, pathogenicity and immune protective effects as an inactivated vaccine candidate. Virus Res 2019; 259:18–27 [View Article] [PubMed]
    [Google Scholar]
  31. Sun J, Li Q, Shao C, Ma Y, He H et al. Isolation and characterization of Chinese porcine epidemic diarrhea virus with novel mutations and deletions in the S gene. Vet Microbiol 2018; 221:81–89 [View Article] [PubMed]
    [Google Scholar]
  32. Khatri M. Porcine epidemic diarrhea virus replication in duck intestinal cell line. Emerg Infect Dis 2015; 21:549–550 [View Article] [PubMed]
    [Google Scholar]
  33. Huan C, Pan H, Fu S, Xu W, Gao Q et al. Characterization and evolution of the coronavirus porcine epidemic diarrhoea virus HLJBY isolated in China. Transbound Emerg Dis 2020; 67:65–79 [View Article] [PubMed]
    [Google Scholar]
  34. Oh J, Lee KW, Choi HW, Lee C. Immunogenicity and protective efficacy of recombinant S1 domain of the porcine epidemic diarrhea virus spike protein. Arch Virol 2014; 159:2977–2987 [View Article] [PubMed]
    [Google Scholar]
  35. Sun D, Feng L, Shi H, Chen J, Cui X et al. Identification of two novel B cell epitopes on porcine epidemic diarrhea virus spike protein. Vet Microbiol 2008; 131:73–81 [View Article] [PubMed]
    [Google Scholar]
  36. Duong BTT, Thao PTP, Hoa NT, Thu HT, Phuoc MH et al. Molecular analysis reveals a distinct subgenogroup of porcine epidemic diarrhea virus in northern Vietnam in 2018-2019. Arch Virol 2022; 167:2337–2346 [View Article] [PubMed]
    [Google Scholar]
  37. Sun Y-G, Li R, Xie S, Qiao S, Li Q et al. Identification of a novel linear B-cell epitope within the collagenase equivalent domain of porcine epidemic diarrhea virus spike glycoprotein. Virus Res 2019; 266:34–42 [View Article] [PubMed]
    [Google Scholar]
  38. Wang L, Byrum B, Zhang Y. New variant of porcine epidemic diarrhea virus, United States, 2014. Emerg Infect Dis 2014; 20:917–919 [View Article] [PubMed]
    [Google Scholar]
  39. Hou Y, Wang Q. Emerging highly virulent porcine epidemic diarrhea virus: molecular mechanisms of attenuation and rational design of live attenuated vaccines. Int J Mol Sci 2019; 20:5478 [View Article] [PubMed]
    [Google Scholar]
  40. Beall A, Yount B, Lin C-M, Hou Y, Wang Q et al. Characterization of a pathogenic full-length cDNA clone and transmission model for porcine epidemic diarrhea virus strain PC22A. mBio 2016; 7:e01451-15 [View Article] [PubMed]
    [Google Scholar]
  41. Hao J, Xue C, He L, Wang Y, Cao Y. Bioinformatics insight into the spike glycoprotein gene of field porcine epidemic diarrhea strains during 2011-2013 in Guangdong, China. Virus Genes 2014; 49:58–67 [View Article] [PubMed]
    [Google Scholar]
  42. Wang X-W, Wang M, Zhan J, Liu Q-Y, Fang L-L et al. Pathogenicity and immunogenicity of a new strain of porcine epidemic diarrhea virus containing a novel deletion in the N gene. Vet Microbiol 2020; 240:108511 [View Article] [PubMed]
    [Google Scholar]
  43. Schumacher L, Chen Q, Fredericks L, Gauger P, Bandrick M et al. Evaluation of the efficacy of an S-INDEL PEDV strain administered to pregnant gilts against a virulent Non-S-INDEL PEDV challenge in newborn piglets. Viruses 2022; 14:1801 [View Article] [PubMed]
    [Google Scholar]
  44. Wang D, Ge X, Chen D, Li J, Cai Y et al. The S gene is necessary but not sufficient for the virulence of porcine epidemic diarrhea virus novel variant strain BJ2011C. J Virol 2018; 92:13 [View Article] [PubMed]
    [Google Scholar]
  45. Lu Y, Huang W, Zhong L, Qin Y, Liu X et al. Comparative characterization and pathogenicity of a novel Porcine Epidemic Diarrhea Virus (PEDV) with a naturally occurring truncated ORF3 gene coinfected with PEDVs possessing an intact ORF3 gene in piglets. Viruses 2021; 13:1562 [View Article] [PubMed]
    [Google Scholar]
  46. Sun M, Yu Z, Luo M, Li B, Pan Z et al. Screening host antiviral proteins under the enhanced immune responses induced by a variant strain of porcine epidemic diarrhea virus. Microbiol Spectr 2022; 10:e0066122 [View Article] [PubMed]
    [Google Scholar]
  47. Yang S, Liu G, Savelkoul HFJ, Jansen CA, Li B. Mini-review: microbiota have potential to prevent PEDV infection by improved intestinal barrier. Front Immunol 2023; 14:1230937 [View Article] [PubMed]
    [Google Scholar]
  48. Duarte M, Tobler K, Bridgen A, Rasschaert D, Ackermann M et al. Sequence analysis of the porcine epidemic diarrhea virus genome between the nucleocapsid and spike protein genes reveals a polymorphic ORF. Virology 1994; 198:466–476 [View Article] [PubMed]
    [Google Scholar]
  49. Fan B, Yu Z, Pang F, Xu X, Zhang B et al. Characterization of a pathogenic full-length cDNA clone of a virulent porcine epidemic diarrhea virus strain AH2012/12 in China. Virology 2017; 500:50–61 [View Article] [PubMed]
    [Google Scholar]
  50. Bi J, Zeng S, Xiao S, Chen H, Fang L. Complete genome sequence of porcine epidemic diarrhea virus strain AJ1102 isolated from a suckling piglet with acute diarrhea in China. J Virol 2012; 86:10910–10911 [View Article] [PubMed]
    [Google Scholar]
  51. Zhao W, Yao Y, Zhang Y, Huang B, Tian K. Full-length genome sequence of porcine epidemic diarrhea virus strain SDTA13-2020. Microbiol Resour Announc 2023; 12:e0001423 [View Article] [PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/jgv.0.001953
Loading
/content/journal/jgv/10.1099/jgv.0.001953
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error