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Indian journal of virology : an official organ of Indian Virological Society2013; 24(2); 256-264; doi: 10.1007/s13337-013-0137-0

Genetic Analysis of the Neuraminidase (NA) Gene of Equine Influenza Virus (H3N8) from Epizootic of 2008-2009 in India.

Abstract: The neuraminidase (NA) gene sequences of four Indian equine influenza viruses (EIVs) isolated from epizootic in 2008 and 2009 were analyzed. The phylogenetic relationship and selection pressure of NA genes were established in comparison to other EIVs circulating worldwide along with the domains and motifs of the encoded protein to find out the significance of mutational changes. Among Indian isolates, two amino acid (aa) changes each in Mysore/12/08 (Asn67Tyr & Asp396Gly), Gopeshwar/1/09 (Ile49Val & Asp396Gly), and Uttarkashi/1/09 (Ile49Val & Asp396Gly) isolates were observed in respect to Jammu-Katra/06/08 isolate. Amino acid (aa) sequence analysis also revealed five consistent aa residue changes viz, Gly/Arg40Glu, Tyr66His, Val191Ile, Val209Ile and Asp235Asn in Asian including Indian isolates, Spain/07 and Spain/09 isolates in comparison to other EIVs circulating worldwide. The topology of the phylogenetic tree revealed that the Indian, Chinese, Mongolian and Kazakhstan isolates together formed a subgroup with Yokohama/10 isolate. Spain/07 & Spain/09 isolates showed closest clustering with Asian isolates. This indicates that non-synonymous mutations in Asian isolates with temporal pattern originating from Spain/07, led to the subgroup of the Asian isolates within Florida clade 2 sublineage. The analysis of the predicted secondary structure has not shown any significant difference in the NA proteins of all Indian isolates. Fixed-effects likelihood (FEL) analysis of the selection pressure revealed three codons (43, 355 & 434) under positive selection pressure. The overall evolutionary changes (ω value) of 3.4 indicates NA gene to be under strong selection pressure. Further, seven putative N-glycosylation sites were observed in the NA protein. The mapping of specific aa changes, their mutational and functional analysis need to be carried out to ascertain their role in pathogenecity of the virus.
Publication Date: 2013-05-24 PubMed ID: 24426284PubMed Central: PMC3784901DOI: 10.1007/s13337-013-0137-0Google Scholar: Lookup
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  • Journal Article

Summary

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The research article is dealing with the analysis of genetic changes in the neuraminidase (NA) gene of horse flu viruses found in India during a widespread outbreak in 2008 and 2009.

Objective and Methodology

  • The study focuses on the genetic analysis of the neuraminidase (NA) gene sequences of four equine influenza viruses (EIV) isolated from an epidemic in 2008 and 2009 in India.
  • The researchers establish the phylogenetic relationships and selection pressures of these NA genes by comparing them with other EIVs around the world. The purpose of this is to discover the effect of mutational changes.

Findings

  • The research identified various amino acid changes among the Indian isolates compared to a specific isolate known as Jammu-Katra/06/08.
  • A broader sequence analysis across all EIVs showed five consistent amino acid changes in Asian (including Indian), Spanish ’07 and ’09 isolates.
  • The phylogenetic tree (a branching diagram showing the evolutionary relationships among various species) showed that the studied Indian viruses, together with the ones from China, Mongolia, and Kazakhstan, formed a subgroup with Yokohama/10 isolate. The Spanish ’07 and ’09 isolates showed a close relationship with the Asian group.

Implications and Conclusions

  • The study indicates that particular mutations in Asian isolates, originating from Spain’s 2007 virus, led to the unique subgroup within the Florida clade 2 sublineage.
  • Despite the various mutations, there was no significant difference in the predicted secondary structure of the NA proteins in all Indian isolates. This suggests that these mutations may not drastically affect the overall organization and function of the protein.
  • The furthers study highlighted certain selection pressures causing positive changes in specific codons (molecular units) in the genes. The research discovered a significant overall evolutionary rate (ω value), suggesting these genes are under strong selection pressure, driving adaptation and evolution.
  • The study also found multiple spots in the NA protein where glycosylation (the addition of sugars to a protein) may occur. This addition could potentially influence the function or stability of the protein.
  • However, to ascertain the exact role of these specific amino acid changes in the pathogenicity (disease-causing ability) of the horse flu virus, further research needs to be conducted by mapping these amino acid changes and performing functional and mutational analyses.

Cite This Article

APA
Bera BC, Virmani N, Shanmugasundaram K, Vaid RK, Singh BK, Gulati BR, Anand T, Barua S, Malik P, Singh RK. (2013). Genetic Analysis of the Neuraminidase (NA) Gene of Equine Influenza Virus (H3N8) from Epizootic of 2008-2009 in India. Indian J Virol, 24(2), 256-264. https://doi.org/10.1007/s13337-013-0137-0

Publication

ISSN: 0970-2822
NlmUniqueID: 8610330
Country: India
Language: English
Volume: 24
Issue: 2
Pages: 256-264

Researcher Affiliations

Bera, B C
  • National Research Centre on Equines, Sirsa Road, Hisar, 125001 Haryana India.
Virmani, N
  • National Research Centre on Equines, Sirsa Road, Hisar, 125001 Haryana India.
Shanmugasundaram, K
  • National Research Centre on Equines, Sirsa Road, Hisar, 125001 Haryana India.
Vaid, R K
  • National Research Centre on Equines, Sirsa Road, Hisar, 125001 Haryana India.
Singh, B K
  • National Research Centre on Equines, Sirsa Road, Hisar, 125001 Haryana India.
Gulati, B R
  • National Research Centre on Equines, Sirsa Road, Hisar, 125001 Haryana India.
Anand, T
  • National Research Centre on Equines, Sirsa Road, Hisar, 125001 Haryana India.
Barua, S
  • National Research Centre on Equines, Sirsa Road, Hisar, 125001 Haryana India.
Malik, P
  • National Research Centre on Equines, Sirsa Road, Hisar, 125001 Haryana India.
Singh, R K
  • National Research Centre on Equines, Sirsa Road, Hisar, 125001 Haryana India.

References

This article includes 34 references
  1. Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs.. Nucleic Acids Res 1997 Sep 1;25(17):3389-402.
    doi: 10.1093/nar/25.17.3389pmc: PMC146917pubmed: 9254694google scholar: lookup
  2. Bean WJ, Schell M, Katz J, Kawaoka Y, Naeve C, Gorman O, Webster RG. Evolution of the H3 influenza virus hemagglutinin from human and nonhuman hosts.. J Virol 1992 Feb;66(2):1129-38.
  3. Bryant NA, Rash AS, Russell CA, Ross J, Cooke A, Bowman S, MacRae S, Lewis NS, Paillot R, Zanoni R, Meier H, Griffiths LA, Daly JM, Tiwari A, Chambers TM, Newton JR, Elton DM. Antigenic and genetic variations in European and North American equine influenza virus strains (H3N8) isolated from 2006 to 2007.. Vet Microbiol 2009 Jul 2;138(1-2):41-52.
    doi: 10.1016/j.vetmic.2009.03.004pubmed: 19346084google scholar: lookup
  4. Crawford PC, Dubovi EJ, Castleman WL, Stephenson I, Gibbs EP, Chen L, Smith C, Hill RC, Ferro P, Pompey J, Bright RA, Medina MJ, Johnson CM, Olsen CW, Cox NJ, Klimov AI, Katz JM, Donis RO. Transmission of equine influenza virus to dogs.. Science 2005 Oct 21;310(5747):482-5.
    doi: 10.1126/science.1117950pubmed: 16186182google scholar: lookup
  5. Daly JM, Lai AC, Binns MM, Chambers TM, Barrandeguy M, Mumford JA. Antigenic and genetic evolution of equine H3N8 influenza A viruses.. J Gen Virol 1996 Apr;77 ( Pt 4):661-71.
    doi: 10.1099/0022-1317-77-4-661pubmed: 8627254google scholar: lookup
  6. Felsenstein J. CONFIDENCE LIMITS ON PHYLOGENIES: AN APPROACH USING THE BOOTSTRAP.. Evolution 1985 Jul;39(4):783-791.
    doi: 10.2307/2408678pubmed: 28561359google scholar: lookup
  7. Hughes MT, Matrosovich M, Rodgers ME, McGregor M, Kawaoka Y. Influenza A viruses lacking sialidase activity can undergo multiple cycles of replication in cell culture, eggs, or mice.. J Virol 2000 Jun;74(11):5206-12.
  8. Kirkland PD, Finlaison DS, Crispe E, Hurt AC. Influenza virus transmission from horses to dogs, Australia.. Emerg Infect Dis 2010 Apr;16(4):699-702.
    doi: 10.3201/eid1604.091489pmc: PMC3321956pubmed: 20350392google scholar: lookup
  9. Lai AC, Chambers TM, Holland RE Jr, Morley PS, Haines DM, Townsend HG, Barrandeguy M. Diverged evolution of recent equine-2 influenza (H3N8) viruses in the Western Hemisphere.. Arch Virol 2001;146(6):1063-74.
    doi: 10.1007/s007050170106pubmed: 11504416google scholar: lookup
  10. Li S, Schulman J, Itamura S, Palese P. Glycosylation of neuraminidase determines the neurovirulence of influenza A/WSN/33 virus.. J Virol 1993 Nov;67(11):6667-73.
  11. Matrosovich MN, Matrosovich TY, Gray T, Roberts NA, Klenk HD. Neuraminidase is important for the initiation of influenza virus infection in human airway epithelium.. J Virol 2004 Nov;78(22):12665-7.
  12. Mitnaul LJ, Matrosovich MN, Castrucci MR, Tuzikov AB, Bovin NV, Kobasa D, Kawaoka Y. Balanced hemagglutinin and neuraminidase activities are critical for efficient replication of influenza A virus.. J Virol 2000 Jul;74(13):6015-20.
  13. Müller I, Jaureguiberry B, Valenzuela PD. Isolation, sequencing and phylogenetic analysis of the hemagglutinin, neuraminidase and nucleoprotein genes of the Chilean equine influenza virus subtypes H7N7 and H3N8.. Biol Res 2005;38(1):55-67.
  14. Mumford J, Wood J. WHO/OIE meeting: consultation on newly emerging strains of equine influenza. 18-19 May 1992, Animal Health Trust, Newmarket, Suffolk, UK.. Vaccine 1993;11(11):1172-5.
    doi: 10.1016/0264-410X(93)90092-Cpubmed: 8249440google scholar: lookup
  15. Ohuchi M, Feldmann A, Ohuchi R, Klenk HD. Neuraminidase is essential for fowl plague virus hemagglutinin to show hemagglutinating activity.. Virology 1995 Sep 10;212(1):77-83.
    doi: 10.1006/viro.1995.1455pubmed: 7676651google scholar: lookup
  16. Payungporn S, Crawford PC, Kouo TS, Chen LM, Pompey J, Castleman WL, Dubovi EJ, Katz JM, Donis RO. Influenza A virus (H3N8) in dogs with respiratory disease, Florida.. Emerg Infect Dis 2008 Jun;14(6):902-8.
    doi: 10.3201/eid1406.071270pmc: PMC2600298pubmed: 18507900google scholar: lookup
  17. Qi T, Guo W, Huang W, Dai L, Zhao L, Li H, Li X, Zhang X, Wang Y, Yan Y, He N, Xiang W. Isolation and genetic characterization of H3N8 equine influenza virus from donkeys in China.. Vet Microbiol 2010 Aug 26;144(3-4):455-60.
    doi: 10.1016/j.vetmic.2010.01.006pubmed: 20153940google scholar: lookup
  18. Saito T, Kawano K. Loss of glycosylation at Asn144 alters the substrate preference of the N8 influenza A virus neuraminidase.. J Vet Med Sci 1997 Oct;59(10):923-6.
    doi: 10.1292/jvms.59.923pubmed: 9362042google scholar: lookup
  19. Saito T, Kawaoka Y, Webster RG. Phylogenetic analysis of the N8 neuraminidase gene of influenza A viruses.. Virology 1993 Apr;193(2):868-76.
    doi: 10.1006/viro.1993.1196pubmed: 8460490google scholar: lookup
  20. Saito T, Taylor G, Laver WG, Kawaoka Y, Webster RG. Antigenicity of the N8 influenza A virus neuraminidase: existence of an epitope at the subunit interface of the neuraminidase.. J Virol 1994 Mar;68(3):1790-6.
  21. Pond SL, Frost SD, Muse SV. HyPhy: hypothesis testing using phylogenies.. Bioinformatics 2005 Mar 1;21(5):676-9.
    doi: 10.1093/bioinformatics/bti079pubmed: 15509596google scholar: lookup
  22. Pond SL, Frost SD. Datamonkey: rapid detection of selective pressure on individual sites of codon alignments.. Bioinformatics 2005 May 15;21(10):2531-3.
    doi: 10.1093/bioinformatics/bti320pubmed: 15713735google scholar: lookup
  23. Shtyrya YA, Mochalova LV, Bovin NV. Influenza virus neuraminidase: structure and function.. Acta Naturae 2009 Jul;1(2):26-32.
    pmc: PMC3347517pubmed: 22649600
  24. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods.. Mol Biol Evol 2011 Oct;28(10):2731-9.
    doi: 10.1093/molbev/msr121pmc: PMC3203626pubmed: 21546353google scholar: lookup
  25. Tu J, Zhou H, Jiang T, Li C, Zhang A, Guo X, Zou W, Chen H, Jin M. Isolation and molecular characterization of equine H3N8 influenza viruses from pigs in China.. Arch Virol 2009;154(5):887-90.
    doi: 10.1007/s00705-009-0381-1pubmed: 19396578google scholar: lookup
  26. Uppal PK, Yadav MP. Outbreak of equine influenza in India.. Vet Rec 1987 Dec 12;121(24):569-70.
    pubmed: 2829407
  27. Varghese JN, Colman PM. Three-dimensional structure of the neuraminidase of influenza virus A/Tokyo/3/67 at 2.2 A resolution.. J Mol Biol 1991 Sep 20;221(2):473-86.
    doi: 10.1016/0022-2836(91)80068-6pubmed: 1920428google scholar: lookup
  28. Virmani N, Singh BK, Gulati BR, Kumar S. Equine influenza outbreak in India.. Vet Rec 2008 Nov 15;163(20):607-8.
    doi: 10.1136/vr.163.20.607-apubmed: 19011253google scholar: lookup
  29. Virmani N, Bera BC, Singh BK, Shanmugasundaram K, Gulati BR, Barua S, Vaid RK, Gupta AK, Singh RK. Equine influenza outbreak in India (2008-09): virus isolation, sero-epidemiology and phylogenetic analysis of HA gene.. Vet Microbiol 2010 Jul 14;143(2-4):224-37.
    doi: 10.1016/j.vetmic.2009.12.007pubmed: 20053509google scholar: lookup
  30. Virmani N, Bera BC, Gulati BR, Karuppusamy S, Singh BK, Kumar Vaid R, Kumar S, Kumar R, Malik P, Khurana SK, Singh J, Manuja A, Dedar R, Gupta AK, Yadav SC, Chugh PK, Narwal PS, Thankur VL, Kaul R, Kanani A, Rautmare SS, Singh RK. Descriptive epidemiology of equine influenza in India (2008-2009): temporal and spatial trends.. Vet Ital 2010 Oct-Dec;46(4):449-58.
    pubmed: 21120800
  31. Virmani N, Bera BC, Shanumugasundaram K, Singh BK, Gulati BR, Singh RK, Vaid RK. Genetic analysis of the matrix and non-structural genes of equine influenza virus (H3N8) from epizootic of 2008-2009 in India.. Vet Microbiol 2011 Aug 26;152(1-2):169-75.
    doi: 10.1016/j.vetmic.2011.04.011pubmed: 21620592google scholar: lookup
  32. Wagner R, Wolff T, Herwig A, Pleschka S, Klenk HD. Interdependence of hemagglutinin glycosylation and neuraminidase as regulators of influenza virus growth: a study by reverse genetics.. J Virol 2000 Jul;74(14):6316-23.
  33. Webster RG, Bean WJ, Gorman OT, Chambers TM, Kawaoka Y. Evolution and ecology of influenza A viruses.. Microbiol Rev 1992 Mar;56(1):152-79.
    pmc: PMC372859pubmed: 1579108doi: 10.1128/mr.56.1.152-179.1992google scholar: lookup
  34. Yamanaka T, Niwa H, Tsujimura K, Kondo T, Matsumura T. Epidemic of equine influenza among vaccinated racehorses in Japan in 2007.. J Vet Med Sci 2008 Jun;70(6):623-5.
    doi: 10.1292/jvms.70.623pubmed: 18628606google scholar: lookup

Citations

This article has been cited 3 times.
  1. Miño S, Mojsiejczuk L, Guo W, Zhang H, Qi T, Du C, Zhang X, Wang J, Campos R, Wang X. Equine Influenza Virus in Asia: Phylogeographic Pattern and Molecular Features Reveal Circulation of an Autochthonous Lineage.. J Virol 2019 Jul 1;93(13).
    doi: 10.1128/JVI.00116-19pubmed: 31019053google scholar: lookup
  2. Singh RK, Dhama K, Karthik K, Khandia R, Munjal A, Khurana SK, Chakraborty S, Malik YS, Virmani N, Singh R, Tripathi BN, Munir M, van der Kolk JH. A Comprehensive Review on Equine Influenza Virus: Etiology, Epidemiology, Pathobiology, Advances in Developing Diagnostics, Vaccines, and Control Strategies.. Front Microbiol 2018;9:1941.
    doi: 10.3389/fmicb.2018.01941pubmed: 30237788google scholar: lookup
  3. Favaro PF, Fernandes WR, Reischak D, Brandão PE, Silva SOS, Richtzenhain LJ. Evolution of equine influenza viruses (H3N8) during a Brazilian outbreak, 2015.. Braz J Microbiol 2018 Apr-Jun;49(2):336-346.
    doi: 10.1016/j.bjm.2017.07.003pubmed: 29100932google scholar: lookup