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Volume 25, Number 11—November 2019
Research Letter

Introduction of Avian Influenza A(H6N5) Virus into Asia from North America by Wild Birds

Sol Jeong, Dong-Hun LeeComments to Author , Yu-Jin Kim, Sun-Hak Lee, Andrew Y. Cho, Jin-Yong Noh1, Erdene-Ochir Tseren-Ochir2, Jei-Hyun Jeong, and Chang-Seon SongComments to Author 
Author affiliations: University of Connecticut, Storrs, Connecticut, USA (S. Jeong, D.-H. Lee); Konkuk University, Seoul, South Korea (S. Jeong, Y.-J. Kim, S.-H. Lee, A.Y. Cho, J.-Y. Noh, E.-O. Tseren-Ochir, J.-H. Jeong, C.-S. Song)

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An avian influenza A(H6N5) virus with all 8 segments of North American origin was isolated from wild bird feces in South Korea. Phylogenetic analysis suggests that this virus may have been introduced into Asia by wild birds, highlighting the role of wild birds in the dispersal of these viruses.

Avian influenza viruses (AIVs) have evolved into phylogenetically independent lineages as a result of separated distribution and migration of wild waterfowl, which are the natural reservoir of the viruses, because of natural geographic barriers. However, the ecologic separation of the migrating hosts seems incomplete, as some species of wild waterfowl (e.g., Northern pintails) migrate across the Bering Strait and serve as an intercontinental bridge for AIV between Eurasia and North America (1). Evidence for intercontinental exchange of AIV has been reported more frequently along continental margins where migratory flyways overlap, such as western Alaska (2); previous studies have described the exchange of gene segments or dispersal of complete virus between the 2 continents through migratory bird movements (24). We report detection of a fully North American AIV A(H6N5) subtype from South Korea during 2017.

During September 2017–March 2018, a total of 4,403 fresh fecal samples were collected in wild bird habitats of South Korea as part of an annual surveillance program for AIV. The sampling was focused on waterfowl at freshwater habitats such as rivers, streams, and reservoirs. We screened the samples for influenza A virus by real-time reverse transcription PCR (rRT-PCR) targeting the matrix gene and by embryonated chicken egg inoculation. A total of 131 samples tested positive for the matrix gene rRT-PCR; 58 AIV subtypes were isolated, including 1 H1N1, 6 H1N2, 2 H1N3, 1 H3N8, 4 H5N2, 3 H5N3, 40 highly pathogenic H5N6 (5), and 1 H6N5. We sequenced full-length genomes of the isolates using next-generation sequencing, as described previously (5). We deposited the nucleotide sequences of A/Mandarin duck/Korea/K17-1638-5/2017(H6N5) virus (hereafter 1638-5/2017 virus) in GenBank (accession nos. MK830100–7).

The Gimpo area (37°43′37″N, 126°39′54″E), from where the H6N5 virus was isolated, is one of the major wintering sites of such migratory birds as the greater white-fronted goose (Anser albifrons), bean goose (Anser fabalis), mallard (Anas platyrhynchos), and Eastern spot-billed duck (Anas poecilorhyncha) (Appendix 1 Figure 1). We identified the host of influenza A virus–positive feces using a DNA barcoding technique (5). We conducted comparative phylogenetic analysis of the 1638-5/2017 virus to trace its origin. For each segment, we aligned sequences with most of the closest full-length related sequences obtained from BLAST ( and used these sequences for phylogenetic analysis. We constructed maximum-likelihood phylogenetic trees using the general time-reversible plus gamma substitution model with 1,000 bootstrap replications in RAxML version 8 (6).


Thumbnail of Maximum-likelihood phylogenetic trees indicating relationships between Eurasian (black circles) and North American (white circles) lineages of avian influenza A viruses. Black arrows indicate genome sequences of A/Mandarin duck/Korea/K17-1638–5/2017(H6N5) virus isolated in this study. Bootstrap values >99% are shown. Maximum-likelihood phylogenetic trees with complete strain names are shown in Appendix 1 Figure 2. HA, hemagglutinin gene; M, matrix gene; NA, neuraminidase gene; NP

Figure. Maximum-likelihood phylogenetic trees indicating relationships between Eurasian (black circles) and North American (white circles) lineages of avian influenza A viruses. Black arrows indicate genome sequences of A/Mandarin duck/Korea/K17-1638–5/2017(H6N5) virus isolated in...

Search results in the BLAST database indicated that all 8 viral gene segments of the 1638-5/2017 virus shared >98% nucleotide identity with North American wild bird AIV collected during 2015–2017 (Appendix 1 Table). Only genetic sequences of North American ancestry virus were in the sequencing data from 1638-5/2017 virus, suggesting the absence of contamination of viral genes from Eurasian-lineage virus during the sampling and virus isolation. Consistent with these findings, maximum-likelihood phylogenetic analysis indicated that all 8 gene segments clustered together with the sequences of AIV isolated from North American wild birds during 2016–2017, rather than those of Eurasian isolates (Figure; Appendix 1 Figure 2; Appendix 2). These results strongly suggest the introduction of AIV of North American ancestry into Eurasia by wild birds.

The high nucleotide similarity of each gene segment with North American AIV identified during 2015–2017 suggests that the genome constellation of the 1638-5/2017 virus had been recently dispersed from North America to Eurasia. Mandarin ducks (Aix galericulata) are found mainly in the Far East and Southeast Asia, within the East Asian–Australasian flyway, and are not recognized as making regular movements between Eurasia and North America. The 1638-5/2017 virus might have been introduced from North America into Eurasia by wild birds migrating between Eurasia and North America and then been transmitted to Mandarin ducks.

Repeated reports of intercontinental transmission of AIV, particularly detections of fully Eurasian AIV, including H8N4 (4) and H9N2 (7); H16N3 (8) and highly pathogenic H5Nx (3) in North America; and the fully North American H6N5 virus in South Korea (this study) suggest that dispersal of AIV between Eurasia and North America is bidirectional and might not be exceedingly rare (2,7). Bahl et al. reported that the multiple introductions of Eurasian H6 AIV resulted in the establishment of viral sublineages in the North American AIV gene pool and changed the evolutionary dynamics of AIV in wild birds in North America (9). In addition, the North American N8 subtype gene has been established in wild bird populations migrating through the East Asian–Australasian flyway and was identified from a chicken farm and live bird market in South Korea (10). Enhanced influenza surveillance along migratory flyways and complete genome sequencing of identified viruses would be essential for better understanding of the intercontinental migrations of AIV and for early detection of the introduction of novel strains.

Dr. Jeong is a veterinarian and PhD candidate at Konkuk University, Seoul, South Korea. Her primary research interests include the molecular epidemiology and host–pathogen interactions of avian influenza viruses.



This research was supported by the Bio and Medical Technology Development Program of the National Research Foundation (NRF) funded by the South Korean government (MSIT) (No. NRF-2018M3A9H4056535). D.-H.L. is partially supported by the US Department of Agriculture, Agricultural Research Service project no. 6040-32000-066-51S.



  1. Olsen  B, Munster  VJ, Wallensten  A, Waldenström  J, Osterhaus  AD, Fouchier  RA. Global patterns of influenza A virus in wild birds. Science. 2006;312:3848. DOIPubMedGoogle Scholar
  2. Ramey  AM, Pearce  JM, Flint  PL, Ip  HS, Derksen  DV, Franson  JC, et al. Intercontinental reassortment and genomic variation of low pathogenic avian influenza viruses isolated from northern pintails (Anas acuta) in Alaska: examining the evidence through space and time. Virology. 2010;401:17989. DOIPubMedGoogle Scholar
  3. Lee  DH, Torchetti  MK, Winker  K, Ip  HS, Song  CS, Swayne  DE. Intercontinental spread of Asian-origin H5N8 to North America through Beringia by migratory birds. J Virol. 2015;89:65214. DOIPubMedGoogle Scholar
  4. Ramey  AM, Reeves  AB, Donnelly  T, Poulson  RL, Stallknecht  DE. Introduction of Eurasian-origin influenza A(H8N4) virus into North America by migratory birds. Emerg Infect Dis. 2018;24:19503.
  5. Kwon  JH, Jeong  S, Lee  DH, Swayne  DE, Kim  YJ, Lee  SH, et al. New reassortant clade avian influenza A(H5N6) virus in wild birds, South Korea, 2017–18. Emerg Infect Dis. 2018;24:19535.
  6. Stamatakis  A. RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. Bioinformatics. 2014;30:13123. DOIPubMedGoogle Scholar
  7. Ramey  AM, Reeves  AB, Sonsthagen  SA, TeSlaa  JL, Nashold  S, Donnelly  T, et al. Dispersal of H9N2 influenza A viruses between East Asia and North America by wild birds. Virology. 2015;482:7983. DOIPubMedGoogle Scholar
  8. Huang  Y, Wille  M, Benkaroun  J, Munro  H, Bond  AL, Fifield  DA, et al. Perpetuation and reassortment of gull influenza A viruses in Atlantic North America. Virology. 2014;456-457:35363. DOIPubMedGoogle Scholar
  9. Bahl  J, Vijaykrishna  D, Holmes  EC, Smith  GJ, Guan  Y. Gene flow and competitive exclusion of avian influenza A virus in natural reservoir hosts. Virology. 2009;390:28997. DOIPubMedGoogle Scholar
  10. Lee  DH, Lee  HJ, Lee  YN, Park  JK, Lim  TH, Kim  MS, et al. Evidence of intercontinental transfer of North American lineage avian influenza virus into Korea. Infect Genet Evol. 2011;11:2326. DOIPubMedGoogle Scholar




Cite This Article

DOI: 10.3201/eid2511.190604

Original Publication Date: October 03, 2019

1Current affiliation: Konkuk Ctc bio Animal Vaccine (KCAV) Co., Ltd., Seoul, South Korea.

2Current affiliation: Mongolian University of Life Sciences, Ulaanbaatar, Mongolia.

Table of Contents – Volume 25, Number 11—November 2019

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Please use the form below to submit correspondence to the authors or contact them at the following address:

Dong-Hun Lee, University of Connecticut Department of Pathobiology and Veterinary Science, 61 North Eagleville Road, Storrs, Connecticut, USA; ; Chang-Seon Song, Konkuk University College of Veterinary Medicine, 120 Neungdong-ro, Gwangjin-gu, Seoul 05029, South Korea

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Page created: October 15, 2019
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