The outbreaks of the highly pathogenic avian influenza (HPAI) in 2016-2017 and 2017-2018, caused by novel reassortant clade 2.3.4.4 H5N6 viruses, resulted in the loss of one billion birds in South Korea. Here, we characterized the H5N6 viruses isolated from wild birds in South Korea from December 2017 to August 2019 by next-generation sequencing. The results indicated that clade 2.3.4.4 H5N6 viruses isolated in 2017 and 2019 shared almost identical nucleotide sequences with the HPAI H5N6 viruses from 2016 in South Korea. This repeated detection of evolutionarily identical H5N6 viruses in same region for more than three years may suggest indigenization of the HPAI H5N6 virus in South Korea. Phylogenetic analysis demonstrated that the clade 2.3.4.4 H5N6 viruses isolated in 2017 and 2019 were evolutionarily distinct from those isolated in 2018. Molecular analysis revealed that the H5N6 viruses isolated in 2017 and 2019 had features associated with an increased risk of human infection (e.g. a deletion at position 133 of HA and glutamic acid residue at position 92 of NS1). Overall, these genomic features of HPAI H5N6 viruses highlight the need for continuous monitoring of avian influenza viruses in wild migratory birds as well as in domestic birds. Highly pathogenic avian influenza (HPAI) viruses cause disease in poultry and humans as well as in some other mammals and wild birds, demonstrating a serious threat to the economy and public. As the Asian HPAI H5N1 viruses were first identified from A/goose/Guangdong/1996 (H5N1) in China, the H5 hemagglutinin (HA) gene has evolved into 10 genetically unique clades (0-9) 1. Among them, novel HPAI virus reassortant for the HA gene in the H5 clade 2.3.4.4 with different neuraminidase (NA) subtypes have been isolated in animals and humans worldwide 2 , and are divided into four genetically distinct subgroups, AD , based on phylogenetic analysis 3-6. Group A and Group B comprise H5N8 viruses, which were first identified from China and South Korea in late 2013/early 2014 3. During the winter season of 2017-2018, Group B H5N6 viruses, which had acquired the NA gene of the Eurasian low pathogenic avian influenza virus, were identified from birds in England, Germany, Japan, and Taiwan 4,5. Group C comprises H5N6 viruses identified from China and Laos during 2013-2014, and Group D comprises H5N6 viruses identified from China and Vietnam during 2013-2014 including human strains, albeit they have not been formally adopted 3,6. It is reported that Group B H5N6 viruses differ phylogenetically from Group C H5N6 viruses circulating in China 4. Overall, clade 2.3.4.4 H5N6 viruses have spread rapidly and caused worldwide outbreaks in poultry; 7,8 moreover and 21 laboratory-confirmed human infections have been reported 3. In South Korea, large HPAI outbreaks in domestic poultry occurred during 2014-2018. The outbreaks in the winter seasons of 2016-2017 and 2017-2018, caused by novel reassortant clade 2.3.4.4 H5N6 viruses 5,9-11 , resulted in the loss of one billion birds in 440 farms in South...
This Article contains errors. The authors reports that samples were collected from allantoic fluid; they were collected from fecal samples. The authors also reported incorrect biosafety level for the facilities where the experiments were performed. In Materials and Methods, 'Sample sites and sample collection' , "After HA activity testing, viral RNA was extracted from the HA-positive allantoic fluid using QIAamp Viral RNA Mini Kit (QIAGEN, Hilden, Germany). Approval for this study was obtained from the Institutional Animal Care and Use Committee at Chungcheongbuk-do Veterinary Service Laboratory (#2019-2) and all experimental procedures from virus isolation to sequencing library preparation were conducted in approved biosafety level 3 (BSL3) facilities (KCDC-12-3-04) at Chungbuk Veterinary Service Laboratory. " should read: "After HA activity testing, viral RNA was extracted from the HA-positive fecal samples using QIAamp Viral RNA Mini Kit (QIAGEN, Hilden, Germany). Approval for this study was obtained from the Institutional Animal Care and Use Committee at Chungcheongbuk-do Veterinary Service Laboratory (#2019-2) and all experimental procedures from virus isolation to sequencing library preparation were conducted in approved biosafety level 2 (BSL2) facilities (QIA-GA2-16-015) at Chungbuk Veterinary Service Laboratory joongbu. "
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