Abstract
The low pathogenic avian influenza (LPAI) H9N2 subtype has become the most prevalent and widespread in many Asian and Middle Eastern countries. It causes an enzootic situation in commercial poultry and known as a potential facilitator virus that can be transmitted to human from birds. The neuraminidase (NA) gene plays an important role the release and spread of the virus from infected cells and throughout the bird. The complete nucleotide sequences of the NA gene of seven H9N2 viruses collected from apparent healthy chicken and quail flocks in Egypt during 2014–2015, were amplified and sequenced. The phylogenetic relationships were investigated and all viruses were belonging to the A/Q/HK/G1/97 strain (G1-like). There were no insertions or deletions or shortening in NA stalk regions when compared to Y280-lineage and the human H9N2 isolates. No obvious changes NA interactions with antiviral drugs. We found that the Egyptian H9N2 viruses have seven glycosylation sites like the most recorded H9N2 viruses in the country, except A/Q/Egypt/14864V/2014 virus which has only six. The NA has four amino acid substitutions distributed in different parts of the hemadsorbing site. The most characteristic substitutions in this site were S372A and W403R these substitutions were a distinctive feature resembling to human H9N2, H2N2 and H3N2 viruses but differs from the other avian influenza viruses. These Special features of surface glycoproteins of LPAI-H9N2 viruses refer to the tendency for enhanced introductions into humans and ensuring the importance of poultry in the transfer influenza viruses.
Similar content being viewed by others
References
Aamir UB, Wernery U, Ilyushina N, Webster RG. Characterization of avian H9N2 influenza viruses from United Arab Emirates 2000 to 2003. Virology. 2007;361:45–55.
Air GM, Laver WG, Webster RG. Mechanism of antigenic variation in an individual epitope on influenza virus N9 neuraminidase. J Virol. 1990;64:5797–803.
Alexander DJ. An overview of the epidemiology of avian influenza. Vaccine. 2007;25:5637–44.
Arafa AS, Hagag NM, Yehia N, Zanaty AM, Naguib MM, Nasef SA. Effect of cocirculation of highly pathogenic avian influenza H5N1 subtype with low pathogenic H9N2 subtype on the spread of infections. Avian Dis. 2012;56:849–57.
Bahari P, Pourbakhsh SA, Shoushtari H, Bahmaninejad MA. Molecular characterization of H9N2 avian influenza viruses isolated from vaccinated broiler chickens in northeast Iran. Trop Anim Health Prod. 2015;47:1195–201.
Bantia S, Ghate AA, Ananth SL, Babu YS, Air GM, Walsh GM. Generation and characterization of a mutant of influenza A virus selected with the neuraminidase inhibitor BCX-140. Antimicrob Agents Chemother. 1998;42:801–7.
Biswas PK, Christensen JP, Ahmed S, Barua H, Das A, Rahman MH, et al. Avian influenza outbreaks in chickens, Bangladesh. Emerg Infect Dis. 2008;14:1909–12.
Bloom JD, Gong LI, Baltimore D. Permissive secondary mutations enable the evolution of influenza oseltamivir resistance. Science. 2010;328:1272–5.
Colman PM, Tulip WR, Varghese JN, Tulloch PA, Baker AT, Laver WG, et al. Three-dimensional structures of influenza virus neuraminidase-antibody complexes. Philos Trans R Soc Lond B Biol Sci. 1989;323:511–8.
EMPRES (2015) Animal influenza update. Availabe online at http://empres-i.fao.org/empres-i/home. Accessed 13 Feb 2015
Fereidouni SR, Harder TC, Gaidet N, Ziller M, Hoffmann B, Hammoumi S, et al. Saving resources: avian influenza surveillance using pooled swab samples and reduced reaction volumes in real-time RT-PCR. J Virol Methods. 2012;186:119–25.
Fuller TL, Gilbert M, Martin V, Cappelle J, Hosseini P, Njabo KY, Abdel Aziz S, Xiao X, Daszak P, Smith TB. Predicting hotspots for influenza virus reassortment. Emerg Infect Dis. 2013;19(4):581–8.
Fusaro A, Monne I, Salviato A, Valastro V, Schivo A, Amarin NM, et al. Phylogeography and evolutionary history of reassortant H9N2 viruses with potential human health implications. J Virol. 2011;85:8413–21.
Gomaa MR, Kayed AS, Elabd MA, Zeid DA, Zaki SA, El Rifay AS, et al. Avian influenza A(H5N1) and A(H9N2) seroprevalence and risk factors for infection among Egyptians: a prospective, controlled seroepidemiological study. J Infect Dis 2015;211(9):1399–407.
Guan Y, Shortridge KF, Krauss S, Chin PS, Dyrting KC, Ellis TM, et al. H9N2 influenza viruses possessing H5N1-like internal genomes continue to circulate in poultry in southeastern China. J Virol. 2000;74:9372–80.
Gubareva LV, Robinson MJ, Bethell RC, Webster RG. Catalytic and framework mutations in the neuraminidase active site of influenza viruses that are resistant to 4-guanidino-Neu5Ac2en. J Virol. 1997;71:3385–90.
Gubareva LV, Kaiser L, Hayden FG. Influenza virus neuraminidase inhibitors. Lancet. 2000;355:827–35.
Guo YJ, Krauss S, Senne DA, Mo IP, Lo KS, Xiong XP, et al. Characterization of the pathogenicity of members of the newly established H9N2 influenza virus lineages in Asia. Virology. 2000;267:279–88.
Haghighat-Jahromi M, Asasi K, Nili H, Dadras H, Shooshtari A. Coinfection of avian influenza virus (H9N2 subtype) with infectious bronchitis live vaccine. Arch Virol. 2008;153:651–5.
Hall T. BioEdit version 7.0. 0. Distributed by the author. www.mbio.ncsu.edu/BioEdit/bioedit.html. 2004.
Hausmann J, Kretzschmar E, Garten W, Klenk H-D. N1 neuraminidase of influenza virus A/FPV/Rostock/34 has haemadsorbing activity. J Gen Virol. 1995;76:1719–28.
Homme P, Easterday B. Avian influenza virus infections. I. Characteristics of influenza A/Turkey/Wisconsin/1966 virus. Avian Dis. 1970;1:66–74.
Hulse DJ, Webster RG, Russell RJ, Perez DR. Molecular determinants within the surface proteins involved in the pathogenicity of H5N1 influenza viruses in chickens. J Virol. 2004;78:9954–64.
Infl WA, Manu U. WHO manual on animal influenza diagnosis and surveillance. Geneva: World Health Organization; 2002.
Jang J, Hong SH, Kim IH. Validation of a real-time RT-PCR method to quantify newcastle disease virus (NDV) titer and comparison with other quantifiable methods. J Microbiol Biotechnol. 2011;21:100–8.
Kammon A, Heidari A, Dayhum A, Eldaghayes I, Sharif M, Monne I, et al. Characterization of avian influenza and newcastle disease viruses from poultry in Libya. Avian Dis. 2015;59:422–30.
Kandeil A, El-Shesheny R, Maatouq AM, Moatasim Y, Shehata MM, Bagato O, et al. Genetic and antigenic evolution of H9N2 avian influenza viruses circulating in Egypt between 2011 and 2013. Arch Virol. 2014;159:2861–76.
Khan SU, Anderson BD, Heil GL, Liang S, Gray GC. A systematic review and meta-analysis of the seroprevalence of influenza A (H9N2) infection among humans. J Infect Dis. 2015;212:562–9.
Kobasa D, Rodgers ME, Wells K, Kawaoka Y. Neuraminidase hemadsorption activity, conserved in avian influenza A viruses, does not influence viral replication in ducks. J Virol. 1997;71:6706–13.
Lee D-H, Song C-S. H9N2 avian influenza virus in Korea: evolution and vaccination. Clin Exp Vaccine Res. 2013;2:26–33.
Matrosovich M, Zhou N, Kawaoka Y, Webster R. The surface glycoproteins of H5 influenza viruses isolated from humans, chickens, and wild aquatic birds have distinguishable properties. J Virol. 1999;73:1146–55.
Matrosovich M, Tuzikov A, Bovin N, Gambaryan A, Klimov A, Castrucci MR, et al. Early alterations of the receptor-binding properties of H1, H2, and H3 avian influenza virus hemagglutinins after their introduction into mammals. J Virol. 2000;74:8502–12.
Matrosovich MN, Krauss S, Webster RG. H9N2 influenza A viruses from poultry in Asia have human virus-like receptor specificity. Virology. 2001;281:156–62.
Mitnaul LJ, Matrosovich MN, Castrucci MR, Tuzikov AB, Bovin NV, Kobasa D, et al. Balanced hemagglutinin and neuraminidase activities are critical for efficient replication of influenza A virus. J Virol. 2000;74:6015–20.
Monne I, Ormelli S, Salviato A, De Battisti C, Bettini F, Salomoni A, et al. Development and validation of a one-step real-time PCR assay for simultaneous detection of subtype H5, H7, and H9 avian influenza viruses. J Clin Microbiol. 2008;46:1769–73.
Monne I, Hussein HA, Fusaro A, Valastro V, Hamoud MM, Khalefa RA, et al. H9N2 influenza A virus circulates in H5N1 endemically infected poultry population in Egypt. Influenza Other Respir Viruses. 2013;7:240–3.
Moscona A. Neuraminidase inhibitors for influenza. N Engl J Med. 2005;353:1363–73.
Naguib MM, Arafa A-SA, El-Kady MF, Selim AA, Gunalan V, Maurer-Stroh S, et al. Evolutionary trajectories and diagnostic challenges of potentially zoonotic avian influenza viruses H5N1 and H9N2 co-circulating in Egypt. Infect Genet Evol. 2015;34:278–91.
Organization WH. Antigenic and genetic characteristics of zoonotic influenza viruses and development of candidate vaccine viruses for pandemic preparedness. Geneva: World Health Organization; 2014.
Peiris M, Yuen K, Leung C, Chan K, Ip P, Lai R, et al. Human infection with influenza H9N2. Lancet. 1999;354:916–7.
Pond SLK, Muse SV. HyPhy: hypothesis testing using phylogenies. In: Statistical methods in molecular evolution. New York: Springer; 2005. p. 125–81.
Ramneek Mitchell NL, McFarlane RG. Rapid detection and characterisation of infectious bronchitis virus (IBV) from New Zealand using RT-PCR and sequence analysis. N Z Vet J. 2005;53:457–61.
Rashid S, Naeem K, Ahmed Z, Saddique N, Abbas M, Malik S. Multiplex polymerase chain reaction for the detection and differentiation of avian influenza viruses and other poultry respiratory pathogens. Poult Sci. 2009;88:2526–31.
Setiawaty V, Dharmayanti NL, Misriyah Pawestri HA, Azhar M, Tallis G, et al. Avian influenza A(H5N1) virus outbreak investigation: application of the FAO-OIE-WHO four-way linking framework in Indonesia. Zoonoses Publ Health. 2015;62:381–7.
Spackman E, Senne DA, Myers TJ, Bulaga LL, Garber LP, Perdue ML, et al. Development of a real-time reverse transcriptase PCR assay for type A influenza virus and the avian H5 and H7 hemagglutinin subtypes. J Clin Microbiol. 2002;40:3256–60.
Tai CY, Escarpe PA, Sidwell RW, Williams MA, Lew W, Wu H, et al. Characterization of human influenza virus variants selected in vitro in the presence of the neuraminidase inhibitor GS 4071. Antimicrob Agents Chemother. 1998;42:3234–41.
Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol. 2013;30:2725–9.
Varghese JN, Colman PM. Three-dimensional structure of the neuraminidase of influenza virus A/Tokyo/3/67 at 2.2 Å resolution. J Mol Biol. 1991;221:473–86.
Varghese JN, Colman PM, van Donkelaar A, Blick TJ, Sahasrabudhe A, McKimm-Breschkin JL. Structural evidence for a second sialic acid binding site in avian influenza virus neuraminidases. Proc Natl Acad Sci USA. 1997;94:11808–12.
von Itzstein M. The war against influenza: discovery and development of sialidase inhibitors. Nat Rev Drug Discov. 2007;6:967–74.
Webster RG, Bean WJ, Gorman OT, Chambers TM, Kawaoka Y. Evolution and ecology of influenza A viruses. Microbiol Rev. 1992;56:152–79.
Wiley DC, Skehel JJ. The structure and function of the hemagglutinin membrane glycoprotein of influenza virus. Annu Rev Biochem. 1987;56:365–94.
Wong SS, Yuen K-Y. Avian influenza virus infections in humans. Chest J. 2006;129:156–68.
Xu KM, Li KS, Smith GJ, Li JW, Tai H, Zhang JX, et al. Evolution and molecular epidemiology of H9N2 influenza A viruses from quail in southern China, 2000 to 2005. J Virol. 2007;81:2635–45.
Yen H-L, McKimm-Breschkin J, Choy K-T, Wong D, Cheung P, Zhou J, et al. Resistance to neuraminidase inhibitors conferred by an R292K mutation in a human influenza virus H7N9 isolate can be masked by a mixed R/K viral population. MBio. 2013;4:e00396–13.
Zhang K, Zhang Z, Yu Z, Li L, Cheng K, Wang T, et al. Domestic cats and dogs are susceptible to H9N2 avian influenza virus. Virus Res. 2013;175:52–7.
Zürcher T, Yates PJ, Daly J, Sahasrabudhe A, Walters M, Dash L, et al. Mutations conferring zanamivir resistance in human influenza virus N2 neuraminidases compromise virus fitness and are not stably maintained in vitro. J Antimicrob Chemother. 2006;58:723–32.
Acknowledgements
Thanks to the entire gene analysis unit staff, National Laboratory for Quality Control on Poultry Production (NLQP), Animal Health Research Institute, Cairo, Egypt for their great help during the study. Special thanks to Drs. Abdelsatar Arafa, Amany Adel and Naglaa Hagag.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Mosaad, Z., Arafa, A., Hussein, H.A. et al. Mutation signature in neuraminidase gene of avian influenza H9N2/G1 in Egypt. VirusDis. 28, 164–173 (2017). https://doi.org/10.1007/s13337-017-0367-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13337-017-0367-7