Complete Coding Sequences of One H9 and Three H7 Low-Pathogenic Influenza Viruses Circulating in Wild Birds in Belgium, 2009 to 2012.

Steven Van Borm,Annick Linden,Didier Vangeluwe,Sylvie Marché,Mieke Steensels,Toon Rosseel,Bénédicte Lambrecht,Thierry Van Den Berg

doi:10.1128/genomea.00540-16

Abstract

The complete coding sequences of four avian influenza A viruses (two H7N7, one H7N1, and one H9N2) circulating in wild waterfowl in Belgium from 2009 to 2012 were determined using Illumina sequencing. All viral genome segments represent viruses circulating in the Eurasian wild bird population.

Highlights

Virus isolates (1 ml of allantoic fluid from first passage in embryonated chicken eggs) were pretreated by size-selective filtration (0.45 ␮M) and DNase treatment (100 U of DNase I [New England BioLabs] at 37°C for 1 h) before proceeding to total RNA extraction (QIAamp Viral RNA minikit; Qiagen). cDNA was produced using random hexamer primers and SuperScript III reverse transcriptase (Thermo Fisher Scientific), according to the manufacturer’s instructions
During the ongoing active surveillance for avian influenza (AI) in wild birds in Belgium from 2009 to 2012, several AI viruses were isolated from Anseriformes (1)
Two H7N7 viruses, A/Anas platyrhynchos/Belgium/23852cls33/2012 (H7N7) and A/Anas platyrhynchos/Belgium/25576cls41/2012 (H7N7), were isolated from cloacal swabs taken from hunted mallards from different locations in southern Belgium (Sorée on 12 September 2012, and Grand-Leez on 6 November 2012, respectively)

Summary

Introduction

Virus isolates (1 ml of allantoic fluid from first passage in embryonated chicken eggs) were pretreated by size-selective filtration (0.45 ␮M) and DNase treatment (100 U of DNase I [New England BioLabs] at 37°C for 1 h) before proceeding to total RNA extraction (QIAamp Viral RNA minikit; Qiagen). cDNA was produced using random hexamer primers and SuperScript III reverse transcriptase (Thermo Fisher Scientific), according to the manufacturer’s instructions. During the ongoing active surveillance for avian influenza (AI) in wild birds in Belgium from 2009 to 2012, several AI viruses were isolated from Anseriformes (1). We selected four viruses for whole-genome sequencing.

Results

Conclusion