Advertisement

Pathological changes, shedding pattern and cytokines responses in chicks infected with avian influenza-H9N2 and/or infectious bronchitis viruses

  • Osama MahanaEmail author
  • Abdel-Sattar Arafa
  • Ahmed Erfan
  • Hussein A. Hussein
  • Mohamed A. Shalaby
Original Article
  • 19 Downloads

Abstract

Avian influenza H9N2 (AIV-H9N2) and Infectious bronchitis (IB) viruses are the most commonly isolated viruses from poultry flocks suffering from respiratory signs with mortalities. The outcome of co-infection with both viruses hasn’t been yet well understood. In this study, eighty 1-day-old specific pathogen free chicks were divided into four distinct groups. Group 1 remained uninfected as negative control group; groups 2, 3 and 4 were inoculated with either AIV-H9N2 or IBV or co infected with AIV-H9N2 followed by IBV three days post inoculation respectively. Chicks were monitored for clinical and pathological changes, virus shedding and both Interleukin-6 (IL6) and Interferon gamma (IFNγ) cytokines immune responses. Clinical signs varied from mild to moderate respiratory signs in all challenged groups but were more severe in group 4 with mortalities in groups 3 and 4. Tracheal shedding of both viruses washigher in group 4 than group 2 and 3. Mean AIV-H9 virus titer in lung and kidney was higher in group 4 than group 2 in all time points. IFNγ mRNA gene expression in lung was significantly lower in groups3 and 4. In conclusion, this study reports that co-infection of chicks with both viruses enhances the pathogenicity, increases both viruses shedding and extend AIV-H9 replication with impairment of IFNγ stimulation in lung.

Keywords

Avian influenza H9N2 IBV Co-infection Cytokines 

Notes

Acknowledgements

The authors would like to acknowledge Mohamed Ahmed and Mai Morsi for their kind support.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures involving animals were reviewed and approved by the Research committee of the Animal Health Research Institute.

References

  1. 1.
    Abdel-Moneim AS, Afifi MA, El-KadyMF. Emergence of a novel genotype of avian infectious bronchitis virus in Egypt. ArchVirol. 2012;157: 2453–7.  https://doi.org/10.1007/s00705-012-1445. (PMID: 22903394).
  2. 2.
    Asif M, Lowenthal JW, Ford ME, Schat KA, Kimpton WG, Bean AG. Interleukin-6 expression after infectious bronchitis virus infection in chickens. Viral Immunol. 2007;20:479–86.  https://doi.org/10.1089/vim.2006.0109.CrossRefGoogle Scholar
  3. 3.
    Bakaletz LO. Viral potentiation of bacterial superinfection of the respiratory tract. Trendsin Microbiol. 1995;3:110–4.  https://doi.org/10.1016/S0966-842X(00)88892-7.CrossRefGoogle Scholar
  4. 4.
    Bancroft JD, Gamble M (2008) Theory and practices of histologic techniques. Philadelphia, PA: Elsevier; 2008.Google Scholar
  5. 5.
    Ben Shabat M, Meir R, Haddas R, Lapin E, Shkoda I, Raibstein I, Perk S, Davidson I. Development of a real-time TaqMan RT-PCR assay for the detection of H9N2 avian influenza viruses. J Virolog Methods. 2010;168(1–2):72–7.CrossRefGoogle Scholar
  6. 6.
    Cavanagh D, Davis PJ, Cook JK, Li D, Kant A, Koch G. Location of the amino acid differences in the S1 spike glycoprotein subunit of closely related serotypes of infectious bronchitis virus. Avian Pathol. 1992;21:33–43.  https://doi.org/10.1080/03079459208418816. (PMID: 18670913).
  7. 7.
    Capua I, Marangon S. The avian influenza epidemic in Italy, 1999–2000: a review. Avian Pathol. 2000;29:289–94.CrossRefGoogle Scholar
  8. 8.
    Engelich G, White M, Hartshorn KL. Role of the respiratory burst in co-operativereduction in neutrophil survival by influenza A virus and Escherichia coli. J Med Microbiol. 2002;51:484–90.  https://doi.org/10.1099/0022-1317-51-6-484.CrossRefGoogle Scholar
  9. 9.
    Haghighat-Jahromi M, Asasi K, Nili H, Dadras H. Role of infectious bronchitis live vaccine on pathogenicity of H9N2 avian influenza virus. Int J Poult Sci. 2007;6(11):838–841. ISSN 1682-8356.Google Scholar
  10. 10.
    Hassan KE, Shany SA, Ali A, Dahshan AHM, Azza A, El-Kady MF. Prevalence of avian respiratory viruses in broiler flocks in Egypt. Poult Sci. 2016;95:1271–80.  https://doi.org/10.3382/ps/pew068.CrossRefGoogle Scholar
  11. 11.
    Hassan KE, Ali A, Shany Salama AS, El-Kady MF. Experimental co-infection of infectious bronchitis and low pathogenic avian influenza H9N2 viruses in commercial broiler chickens. Res Vet Sci. 2017;115:356–62.CrossRefGoogle Scholar
  12. 12.
    Jang H, Koo B-S, Jeon E-O, Lee H-R, Lee S-M, Mo I-P. Altered pro-inflammatory cytokine mRNA levels in chickens infected with infectious bronchitis virus. Poult Sci. 2013;92:2290–8.  https://doi.org/10.3382/ps.2013-03116.CrossRefGoogle Scholar
  13. 13.
    Kaiser P, Rothwell L, Galyov EE, Barrow PA, Burnside J, Wigley P. Differential cytokine expression in avian cells in response to invasion by Salmonella typhimurium, Salmonella enteritidis and Salmonella gallinarum. Microbiology. 2000;146:3217–26.CrossRefGoogle Scholar
  14. 14.
    Kim KS, Jung H, Shin IK, Choi BR, Kim DH. Induction of interleukin1 beta (IL1β) is a critical component of lung inflammation during influenza A (H1N1) virus infection. J Med Virol. 2015;87:1104–12.  https://doi.org/10.1002/jmv.24138.CrossRefGoogle Scholar
  15. 15.
    Liu J, Li N, Meng D, Hao M, Wei L, Chai T. The mRNA and proteins expression levels analysis of TC-1 cells immune response to H9N2 avian influenza virus. Front Microbiol. 2016;7:1039.  https://doi.org/10.3389/fmicb.2016.01039.Google Scholar
  16. 16.
    Livak JK, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCt method. Methods. 2001;25:402–8.CrossRefGoogle Scholar
  17. 17.
    Meir R, Maharat O, Farnushi Y, Simanov L. Development of a real-time TaqMan® RT-PCR assay for the detection of infectious bronchitis virus in chickens, and comparison of RT-PCR and virus isolation. J Virol Methods. 2010;163:190–4.CrossRefGoogle Scholar
  18. 18.
    Nang NT, Lee JS, Song BM, Kang YM, Kim HS, Seo SH. Induction of inflammatory cytokines and Toll-like receptors in chickens infected with avian H9N2 influenza virus. Vet Res. 2011;42:64.  https://doi.org/10.1186/1297-9716-42-64.CrossRefGoogle Scholar
  19. 19.
    Ng LFP, Liu DX. Further characterization of the coronavirus infectious bronchitis virus C-like proteinase and determination of a new cleavage site. Virology. 2000;272:27–39.CrossRefGoogle Scholar
  20. 20.
    OIE. Chapter 2.3.4. Avian influenza. http://www.oie.int/fileadmin/Home/eng/Health_standards/tahm/2.03.04_AI.pdf (2014). Accessed 22 Nov 2014.
  21. 21.
    Reed LJ, Muench H. A simple method of estimating fifty percent end points. Am J Hyg. 1938;27:493–7.Google Scholar
  22. 22.
    Schroder K, Hertzog PJ, Ravasi T, Hume DA. Interferon-gamma: an overview of signals, mechanisms and functions. J Leukoc Biol. 2004;75:163–89.CrossRefGoogle Scholar
  23. 23.
    Subtain SM, Iqbal CZ, Ahmad AA, Azhar M, Umer S. Study on pathogenesis of low pathogenic avian influenza virus H9 in broiler chickens. Pak J Zool. 2011;43(5):999–1008.Google Scholar
  24. 24.
    Valastro V, Holmes EC, Britton P, Fusaro A, Jackwood MW, Cattoli G, Monne I. S1 gene-based phylogeny of infectious bronchitis virus: an attempt to harmonize virus classification. Infect Genet Evol. 2016;39:349–64.CrossRefGoogle Scholar
  25. 25.
    Vervelde L, Matthijs MG, Van Haarlem DA, et al. Rapid NK-cell activation in chicken after infection with infectious bronchitis virus M41. Vet Immunol Immunopathol. 2013;151:337–41.CrossRefGoogle Scholar
  26. 26.
    Wang J, Cao Z, Guo X, Zhang Y, Wang D, Xu S, Yin Y. Cytokine expression in 3 chicken host systems infected with H9N2 influenza viruses with different pathogenicities. Avian Pathol. 2016;45(6):630–9.CrossRefGoogle Scholar
  27. 27.
    Wood GW, McCauley JW, Bashiruddin JB, Alexander DJ. Deduced amino acid sequences at the hemagglutinin cleavage site of avian influenza A viruses of H5 and H7 subtypes. Arch Virol. 1993;130:209–17.CrossRefGoogle Scholar
  28. 28.
    Yuk SS, Lee DH, Park JK, Tseren-Ochir EO, Kwon JH, Noh JY, Lee JB, Park SY, Choi IS, Song CS. Pre-immune state induced by chickeninterferon gamma inhibits the replicationof H1N1 human and H9N2 avian influenzaviruses in chicken embryo fibroblasts. Virol J. 2016;13:71.  https://doi.org/10.1186/s12985-016-0527-1.CrossRefGoogle Scholar
  29. 29.
    Zanaty A, Arafa A-S, Hagag N, El-Kady M. Genotyping and pathotyping of diversified strains of infectious bronchitis viruses circulating in Egypt. World J Virol. 2016;5(3): 125–134. ISSN 2220-3249.Google Scholar
  30. 30.
    Zhou H, Chen S, Yan B, Chen H, Wang M, Jia R, Zhu D, Liu M, Liu F, Yang Q, Wu Y, Sun K, Chen X, Jing B, Cheng A. LPAIV H9N2 drives the differential expression of goose interferons and proinflammatory cytokines in both in vitro and in vivo studies. Front Microbiol. 2016;7:166.  https://doi.org/10.3389/fmicb.2016.00166.Google Scholar

Copyright information

© Indian Virological Society 2019

Authors and Affiliations

  • Osama Mahana
    • 1
    Email author
  • Abdel-Sattar Arafa
    • 1
  • Ahmed Erfan
    • 1
  • Hussein A. Hussein
    • 2
  • Mohamed A. Shalaby
    • 2
  1. 1.Reference Laboratory for Veterinary Quality Control on Poultry ProductionAnimal Health Research InstituteDokki, GizaEgypt
  2. 2.Deparment of Virology, Faculty of Veterinary MedicineCairo UniversityGizaEgypt

Personalised recommendations