Abstract
To determine the genetic relationship of Iranian viruses, the haemagglutinin (HA) genes from ten isolates of H9N2 viruses isolated from commercial chickens in Iran during 1998–2002 were amplified and sequenced. Sequence analysis and phylogenetic studies were conducted by comparing each isolate with those of the available H9N2 strains at GenBank. All these ten isolates had the same sequence –R-S-S-R/G-L– of proteolytic cleavage site of the HA. Nucleotide sequence comparisons of HA gene from Iranian isolates showed 95.2–99.1% identity within the group. Five isolates had leucine (L) at position 226 instead of glutamine (Q). Phylogenetic analysis showed that all our isolates belonged to the G1-like sublineage. Also these isolates showed some degree of homology with other H9N2 isolates e.g., 94.3–96.9% with qu/HK/G1/97, 96.1–98.6% with pa/Chiba/1/97, 95.6–98.2% with pa/Narita/92A/98, and 94.0–96.3% with HK/1073/99. On the basis of phylogenetic and molecular characterization evidence, we concluded that the H9N2 subtype influenza viruses circulating in chicken flocks in Iran since 1998–2002 had a common origin. The results of this study indicated that all Iranian viruses have the potential to emerge as highly pathogenic influenza virus, and considering the homology of these isolates with human H9N2 strains, it seems that the potential of these avian influenza isolates to infect human should not be overlooked.
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References
Aamir, U.B., Wernery, U., Ilyushina, N., and Webster, R.G., 2007. Characterization of avian H9N2 influenza viruses from United Arab Emirates 2000-2003. Journal of Virology, 361, 45–55.
Alexander, D.J., and Spackman, D., 1981. Characterization of influenza A viruses isolated from turkeys during March–May 1979, Avian Pathology, 10, 281–293.
Banks, J., Speidel, E.C., Harris, P.A., and Alexander, D.J., 2000. Phylogenetic analysis of influenza A viruses of H9 haemagglutinin subtype. Avian Pathology, 29, 353–359.
Brown, I.H., Banks, J., Manvell, R.J., Essen, S.C., Shell, W., Slomka, M., Lond, T., and Alexander, D.J., 2006. Recent epidemiology and ecology of influenza A viruses in avian species in Europe and the Middle East. Developments in Biologicals, 124, 45–50.
Butt, K.M., Smith, G.J., Chen, H., Zhang, L.J., Leung, Y.H., Xu, K.M., Lim, W., Webster, R.G., Yuen, K.Y., Peiris, J.S., and Guan, Y., 2005. Human infection with an avian H9N2 influenza A virus in Hong Kong in 2003. Journal of Clinical Microbiology, 43, 5760–5767.
Cameron, K.R., Gregory, V., Banks, J., Brown, H., Alexander, D.J., Hay, A.J., and Lin, Y.P., 2000. H9N2 subtype influenza A viruses in poultry in Pakistan are closely related to the H9N2 viruses responsible for human infection in Hong Kong. Journal of Virology, 278, 36–41.
Fouchier, R., Munster, V., Wallensten, A., Bestebroer, T.M., Herfst, S., Smith, D., Rimmelzwaan, G.F., Olsen, B., and Osterhaus, A., 2005. Characterization of a novel influenza A virus hemagglutinin subtype (HI6) obtained from black-headed gulls. Journal of Virology, 79, 2814–2822.
Guan, Y., Shortridge, K.F., Krauss, S., and Webster, R.G., 1999. Molecular characterization of H9N2 influenza viruses: were they the donors of the “internal” genes of H5N1 viruses in Hong Kong? Proceeding of the National Academy of Science of the USA, 96, 9363–9367.
Guan, Y., Shortridge, K.F., Krauss, S., Chin, P.S., Dyrting, K.C., Ellis, T.M., Webster, R.G., and Peiris, M., 2000. H9N2 influenza viruses possessing H5N1-like internal genomes continue to circulate in poultry in southeastern china. Journal of Virology, 74, 9372–9380.
Guo, Y.J., Krauss, S., Senne, D.A., Mo, I.P., Lo, K.S., Xiong, X.P., Norwood, M., Shortridge, K.F., Webster, R.G., and Guan, Y., 2000. Characterization of the pathogenicity of members of the newly established H9N2 influenza virus lineage in Asia. Journal of Virology, 267, 279–288.
Ha, Y., Stevens, D.J., Skehel, J.J., and Wiley, D.C., 2001. X-Ray structures of H5 avian and H9 swine influenza virus haemagglutinins bound to avian and human receptor analogs. Proceeding of the National Academy of Science of the USA, 98, 11181–11186.
Haghighat-Jahromi, M., Asasi, K., Nili, H., Dadras, H., and Shoushtari, A.H., 2008. Coinfection of avian influenza virus (H9N2 subtype) with infectious bronchitis live vaccine. Archives of Virology, 153, 651–655.
Hoffmann, E., Stech, J., Guan, Y., Webster, R.G., and Perez, D.R., 2001. Universal primer set for the full-length amplification of all influenza A viruses. Archives of Virology, 146, 2275–2289.
Hongqi, L., Xiufan, L., Jian, C., Daxin, P., Lijun, J., and Young, H., 2003. Phylogenetic analysis of the hemagglutinin genes of twenty-six avian influenza viruses of subtype H9N2 isolated from chickens in china during 1996–2001. Avian Disease, 46, 116–127.
Horimoto, T. and Kawaoka, Y., 2001. Pandemic threat posed by avian influenza A viruses. Clinical Microbiology Reviews 14, 129–149.
Lamb, R.A. and Krug, R.M 2001. Orthomyxoviridae: The viruses and their replication. In: Knip, D.M., Howley, P.M., Chanock, R.M., Melick, J.L., Momath, T.P. and Roizman, B. (Eds.), Fields Virology. (Lippincott-Raven, Philadelphia, PA), 1487–1531.
Mase, M., Imada, T., Sanada, Y., Etoh, M., Sanada, N., Tsukamoto, K., Kawaoka, Y., and Yamaguchi, S., 2001. Imported parakeets harbor H9N2 influenza A viruses that are genetically closely related to those transmitted to humans in Hong Kong. Journal of Virology, 75, 3490–3494.
Matrosovich, M.N., Krauss, S., and Webster, R.G., 2001. H9N2 influenza A viruses from poultry in Asia have human virus-like receptor specificity. Journal of Virology, 281, 156–162.
Mosleh, N., Dadras, H., and Mohammadi, A., 2009. Molecular quantitation of H9N2 avian influenza virus in various organs of broiler chickens using TaqMan real time PCR. Journal of Molecular Genetic Medicine, 3, 152–157.
Nili, H., and Asasi, K., 2003. Avian influenza (H9N2) outbreak in Iran. Avian Disease, 47, 828–831.
Shoushtari, A.H., Habl-Alvarid, M.H., Vascellari, M., and Hedayati, A., 2007. Mortality of wild swans associated with naturally infection with highly pathogenic H5N1 avian influenza virus in Iran. Archives of Razi Institute, 62, 207–213.
Swayne, D.E., Senne, D.A., and Beard, C.W., 1998. Isolation and identification of avian pathogens. 4th Ed., (American Association of Avian Pathologist, Pennsylvania, USA).
Toroghi, R., and Momayez, R., 2006. Biological and molecular characterization of avian influenza virus (H9N2) isolates from Iran. Acta Virologica, 50, 163–8.
Tosh, C., Nagarajan, S., Behera, P., Rajukumar, K., Purohit, V., Kamal, R.P., Murugkar, H.V., Gounalan, S., Pattnaik, B., Vanamayya, P.R., Pradhan, H.K., and Dubey, S.C., 2008. Genetic analysis of H9N2 avian influenza viruses isolated from India. Archives of Virology, 153, 1433–1439.
Verhoeyen, M., Fany, R., Jou, W.M., Devos, R., Huylebroek, D., Saman, E., and Fiers, W., 1980. Antigenic drift between the hemagglutinin of the Hong Kong influenza strain A/Aich/2/68 and A/Victoria/3/75. Nature, 286, 771–776.
Wan, H., and Perez, D.R., 2007. Amino acid 226 in the hemagglutinin of H9N2 influenza viruses determines cell tropism and replication in human air way epithelial cells. Journal of Virology, 81, 5181–5191.
Acknowledgments
We thank Professor Jill Banks and Professor Paul Barrow for their scientific guidance and gratefully acknowledge the excellent technical support provided by Miss Eshratabadi. We are also thankful to Shiraz University for providing part of the financial support.
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Homayounimehr, A.R., Dadras, H., Shoushtari, A. et al. Sequence and phylogenetic analysis of the haemagglutinin genes of H9N2 avian influenza viruses isolated from commercial chickens in Iran. Trop Anim Health Prod 42, 1291–1297 (2010). https://doi.org/10.1007/s11250-010-9565-7
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DOI: https://doi.org/10.1007/s11250-010-9565-7