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New Introduction of Clade 2.3.2.1 Avian Influenza Virus (h5n1) Into Bangladesh

M. R. Islam
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh, Bangladesh

P. Monoura
National Reference Laboratory for Avian Influenza, Bangladesh Livestock Research Institute, Savar, Dhaka, Bangladesh

M. M. Rahman
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh, Bangladesh

M. Nooruzzaman
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh, Bangladesh

R. Parvin
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh, Bangladesh

M. A. Samad
National Reference Laboratory for Avian Influenza, Bangladesh Livestock Research Institute, Savar, Dhaka, Bangladesh

E. H. Chowdhury
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh, Bangladesh

M. E. Haque
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh, Bangladesh

M. Giasuddin
Principal Scientific Officer
National Reference Laboratory for Avian Influenza, Bangladesh Livestock Research Institute, Savar, Dhaka, Bangladesh

Abstract/ Executive Summary:

Since the first outbreak of highly pathogenic H5N1 avian inafluenza (HPAI) in Bangladesh in February 2007, a total of 519 disease events have been reported till 22 October 2011. Partial HA gene sequences of 11 selected H5N1 HPAI isolates of 2007 to 2011 were determined and subjected to phylogenetic analysis. The study revealed a recent introduction of clade 2.3.2 and 2.3.4 viruses into Bangladesh in 2011 in addition to clade 2.2 viruses that had been in circulation since 2007. Clade 2.3.2 virus isolates from Bangladesh are phylogenetically related to the newly designated clade 2.3.2.1 viruses, reported recently from Asia and Eastern Europe.

Keywords: Avian Influenza Virus, Bangladesh, Highly Pathogenic, Genetic Characterization, Phylogeny

Location: Bangladesh Livestock Research Institute

Start Date: 01-01-2007

End Date: 31-12-2011

Program Area: Animal Health and Management

Commodity: Poultry

Objectives:

1. Partial HA gene sequences of 11 selected H5N1 HPAI isolates of 2007 to 2011 were determined and subjected to phylogenetic analysis.

Methodology:

A total of 11 selected H5N1 HPAI virus isolates of Bangladesh, obtained at different time points, were propagated in embryonated chicken eggs. These included isolates from several chickens, a crow, a duck and a quail. The identity of the virus isolates in the allantoic fluid was re-confirmed by RT-PCR for H5 and N1 genes. A primer pair (H5- 155F: 5¢-ACA CAT GCY CAR GAC ATA CT-3¢ and H5- 699R: 5¢-CTY TGR TTY AGT GTT GAT GT-3¢) was used to amplify a 545-bp fragment from the H5 gene. For the detection of N1 gene, the primer pair (N1-580-607F: 5¢-TGA AGT ACA ATG GCA TAA TAA CWG ACA C-3¢ and N1-891-918R: 5¢-CAC TGC ATA TAT ATC CTA TTT GAT ACT CC-3¢) suggested by WHO (WHO, 2007) was used, which amplified a 343-bp fragment. RNA was extracted with Qiagen RNeasy Extraction Kit, and RT-PCR was performed with Qiagen One Step RT-PCR Kit (Hilden, Germany). The following thermal profile was followed for both the reactions: reverse transcription at 50C for 30 min, initial denaturation at 95C for 15 min and then 35 cycles of denaturation at 94⁰C for 60 s, annealing at 50⁰C for 60 s and elongation at 72⁰C for 2 min, and final elongation at 72⁰C for 7 min. The RT-PCR products obtained from the H5 gene of the 11 isolates were sequenced. For phylogenetic analysis, nucleotide sequences of HA gene of these 11 isolates were subjected to multiple alignment by Clustal V method using Lasergene DNAStar software. HA gene sequences representing each clade (WHO/ OIE/FAO, 2008) were downloaded from the GenBank and GISAID databases and included in the phylogenetic analysis. In addition, HA gene sequences of few more selected strains from the People’s Republic of China, Vietnam, Lao, Korea, Japan, Mongolia, Russia and Nepal were also included in the analysis.

Source of Information: Transboundary and Emerging Diseases, 12/2011; 59(5):460-3; doi:10.1111/j.1865-1682.2011.01297.x

Article Link (Online Journal): doi:10.1111/j.1865-1682.2011.01297.x

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Clade 2.3.2 virus isolates from Bangladesh are phylogenetically related to the newly designated clade 2.3.2.1 viruses, reported recently from Asia and Eastern Europe

Knowledge Management: Journal