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

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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.

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Acknowledgments

The authors are thankful to the Indian Council of Agricultural Research for financial assistance and providing the necessary facilities to carry out the work. The authors also wish to thank Sh. Mukesh Chand for the technical assistance, and Sh. Subhas and Sh. Suresh Kumar for assistance in sample collection.

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  1. National Research Centre on Equines, Sirsa Road, Hisar, 125001, Haryana, India

    B. C. Bera, N. Virmani, K. Shanmugasundaram, R. K. Vaid, B. K. Singh, B. R. Gulati, T. Anand, S. Barua, P. Malik & R. K. Singh

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  1. B. C. Bera
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  2. N. Virmani
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Correspondence to R. K. Singh.

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Bera, B.C., Virmani, N., Shanmugasundaram, K. et al. Genetic Analysis of the Neuraminidase (NA) Gene of Equine Influenza Virus (H3N8) from Epizootic of 2008–2009 in India. Indian J. Virol. 24, 256–264 (2013). https://doi.org/10.1007/s13337-013-0137-0

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