Printer Friendly

Highly pathogenic avian influenza virus (H5N1) in domestic poultry and relationship with migratory birds, South Korea.

During the 2006-2007 winter season in South Korea, several outbreaks of highly pathogenic avian influenza virus (H5N1) were confirmed among domestic poultry and in migratory bird habitats. Phylogenetic analysis showed that all isolates were closely related and that all belong to the A/ bar-headed goose/Qinghai/5/2005-like lineage rather than the A/chicken/Korea/ES/2003--like lineage.


Highly pathogenic avian influenza (HPAI) virus (H5N1) has been detected repeatedly in domestic poultry and wild birds since 1997, and it poses a substantial threat to human health (1,2). Since the end of 2003, influenza virus (HhN1) strains have spread in an unprecedented manner in many Asian countries, and the outbreaks have resulted in >170 human deaths in Thailand, Vietnam, Cambodia, and Indonesia. These outbreaks have also caused serious economic losses in the poultry industry (, including South Korea in 2003--the first official report of subtype H5N1 in South Korea's history (3).

Since the outbreak caused by subtype H5N1 from migratory waterfowl on Qinghai Lake (QH) in May 2005 (4,5), outbreaks of QH-like avian influenza virus (H5N1) have been reported in the People's Republic of China, Mongolia, Russia, Europe, and Africa, and have been ascribed to the migration of wild birds (6,7). In contrast to virus found in countries on the western side of Qinghai Lake, the Fujian-like avian influenza virus (H5N1) sublineage has predominated in southern China since late 2005 (8), but no outbreaks were reported in Far-Eastern Asian countries such as South Korea and Japan until October 2006. Eventually, in November 2006 and January 2007, outbreaks of HPAI (H5N1) occurred in South Korea and Japan. Here we report the second outbreak of HPAI (HhN1) among poultry in South Korea since November 2006 and its relationship with 2 HPAI virus (H5N1) strains isolated from migratory bird habitats (i.e., in the environment).

The Study

All of the virus strains from domestic poultry used in this study were isolated by the Korean National Veterinary Research and Quarantine Service (NVRQS) in embryonated eggs that were inoculated with tissues and swab specimens collected from the oropharynx and cloaca of affected birds. Two subtype H5N1 strains were isolated in embryonated eggs that had been inoculated with fecal specimens obtained from migratory bird habitats by Chungbuk National University and Chungnam National University. Viral genes were sequenced and analyzed as described (9). The full-length sequences for each segment were used in phylogenetic analyses. Gene sequences determined in this study have been deposited in GenBank under accession nos. EU233675-EU233746.

On November 22, 2006, NVRQS confirmed the first case of HPAI (HhN1) at a chicken farm in Iksan, Jeollabuk-Do, in South Korea. The affected flock contained 6,500 chickens and had shown a sudden increase in severe clinical signs and high mortality rate (86%), as reported by farmers and veterinarians (Table). During intensive surveillance in December 2006 within a 10-km radius (the surveillance zone) from the first outbreak farms, we found other HPAI (H5N1)--affected farms at Iksan (chicken farm, 3.4% deaths) and Gimje (quail farm, 4% deaths), Jeollabuk-Do (Figure 1).

On December 21, 2006, the fourth outbreak of HPAI (HhN1) was confirmed in Asan, Chungcheongnam-Do, in breeder ducks that had shown a severe drop in egg production but no deaths. During intensive observation within the surveillance zone from the fourth outbreak farm, another HPAI (HhN1) outbreak was confirmed in Cheonan, Chungcheongnam-Do, on January 20, 2007, in a layer chicken farm with a 1% mortality rate (Figure 1). Compared with the rates in the first outbreak, the mortality rates in the more recent outbreaks were low (1%-4%) at notification time. This low proportion of deaths could be attributed to the early reporting system between the farmers and NVRQS, when the mortality rate reached [approximately equal to] 1% of the flock on poultry farms, and to the culling of flocks on reverse transcription--PCR confirmation (usually within 1 day) to prevent the spread of the disease. This could have limited the recorded observations when the infecting influenza virus was eliminated before the full extent of its pathogenicity could be manifested, usually after several days of infection.


Great interest has been focused on the role of migratory birds in the spread of H5N1 subtype and the exchange of virus strains between domestic and wild birds in Asia. Therefore, we surveyed avian influenza virus in migratory birds in South Korea to investigate whether the HPAI (H5N1) outbreaks in domestic poultry bore any relationship to bird migration in the same region. During our routine survey for influenza activity in migratory bird habitats, on December 21, 2006, 2 distinct subtype H5N1 strains were isolated from fecal samples from 2 migratory bird habitats one near the first outbreak farm in Chungcheongnam-Do, and the other from a stream in Chungcheongbuk-Do (Figure 1).

Our phylogenetic analysis of the hemagglutinin (HA) genes of all Korean isolates showed that the isolates belong to the A/bar-headed goose/QH/65/2005 (QH/2005) like lineage that caused an outbreak among wild birds at Qinghai Lake in China during 2005, rather than the first HPAI (H5N1) lineage (A/chicken/Korea/ES/2003) that infected farms in Korea in 2003 (Figure 2). Notably, the 2 isolates from migratory bird habitats were closely related to the H5N1 subtype poultry virus strains: A/environment/Korea/W149/2006 was similar to the viruses that occurred in Chungcheongnam-Do, and A/environment/ Korea/W 150/2006 was similar to viruses that affected birds in Jeollabuk-Do. However, all H5N1 subtype virus strains have a series of basic amino acids at the HA cleavage site (PQGERRRKKR/G), which is a characteristic of influenza viruses that are highly pathogenic to chickens (4,5). The intravenous pathogenicity index score of A/chicken/Korea/ IS/2006 was 3.0 in chickens.


Phylogenetic analysis of the other genes showed a similar evolutionary pattern to the HA gene tree. All 20062007 isolates had a 20-aa deletion in the stalk region (residues 49-68) ofneuraminidase (NA) compared with the NA of A/goose/Guangdong/l/96. Analysis of the raw sequencing traces showed no mutations in NA genes of all isolates associated with resistance to NA inhibitors. All 2006-2007 Korean HPAI (H5N1) isolates had glutamic acid at position 92 of the nonstructural protein 1, a position that is related to the ability of H5N1/97 virus to escape the host antiviral cytokine response (12), and lysine at position 627 in the PB2 protein, which is commonly observed in QH viruses (6). Lysine 627 in PB2 is conserved in authentic human influenza viruses and is associated with high virulence of influenza virus (H5N1) strains in mice (13). Analysis of membrane (M) 2 protein sequences showed that none of the 2006-2007 HPAI (H5N1) Korean strains were resistant to amantadine.


In contrast to the 2003 H5NI subtype isolates (3), all of the 2006-2007 H5N1 subtype isolates were QH/05-like strains. Phylogenetic analysis showed that this sublineage spread wildly through Africa and Europe but not in eastern Asia, until the outbreak reported here. We cannot conclude whether wild migratory birds were the origin of the HPAI virus (H5N1) found in poultry or vice versa in South Korea, because the environmental isolates were obtained after the poultry outbreaks. However, the report of outbreaks of similar HPAI virus (H5N1) strains in Japan on January 13, 2007 ( suggested that migratory birds could be a strong mediator for the spread of HPAI virus (H5N1) in South Korea and Japan, as occurred in 2003, because these 2 countries share similar wild bird migration routes. Therefore, continued monitoring of the domestic and wild bird populations is needed to better understand interspecies transmission and to clarify the importance of avian hosts in the ecology of influenza viruses.

All material published in Emerging Infectious Diseases is in the public domain and may be used and reprinted without special permission; proper citation, however, is required.


We thank Hyuk-Man Kwon, Eun-Im Yun, Kyeong-Hwa Han, Hee-Kyung Jang, and Won-Joung Kim for excellent technical assistance. We also thank the Animal Disease Control Division and the Veterinary Epidemiology Division of NVRQS for their efforts to control HPAI.

This work was supported by grant no. RO1-2005-00010585-0 from the Basic Research Program of the Korea Science and Engineering Foundation and by grant no. C-AD15-2006-1401 from the Research Program of NVRQS.


(1.) Claas EC, Osterhaus AD, van Beek R, De Jong JC, Rimmelzwaan GF, Senne DA, et al. Human influenza A H5N1 virus related to a highly pathogenic avian influenza virus. Lancet. 1998;351:472-7.

(2.) Peiris JS, Yu WC, Leung CW, Cheung CY, Ng WF, Nicholls JM, et al. Re-emergence of fatal human influenza A subtype H5N1 disease. Lancet. 2004;363:617-9.

(3.) Lee CW, Suarez DL, Tumpey TM, Sung HW, Kwon YK, Lee YJ, et al. Characterization of highly pathogenic H5N1 avian influenza A viruses isolated from South Korea. J Virol. 2005;79:3692-702.

(4.) Chen H, Smith GJ, Zhang SY, Qin K, Wang J, Li KS, et al. Avian flu: H5N1 virus outbreak in migratory waterfowl. Nature. 2005;436: 191-2.

(5.) Liu J, Xiao H, Lei K Zhu Q, Qin K, Zhang XW, et al. Highly pathogenic H5N1 influenza virus infection in migratory birds. Science. 2005;309:1206.

(6.) Chen H, Li Y, Li Z, Shi J, Shinya K, Deng G, et al. Properties and dissemination of HSN1 viruses isolated during an influenza outbreak in migratory waterfowl in western China. J Virol. 2006;80: 5976-83.

(7.) Olsen B, Munster V J, Wallensten A, Waldenstrom J, Osterhaus AD, Fouchier RA. Global patterns of influenza a virus in wild birds. Science. 2006;312:384-8.

(8.) Smith GJ, Fan XH, Wang J, Li KS, Qin K, Zhang JX, et al. Emergence and predominance of an H5N1 influenza variant in China. Proc Natl Acad Sci U S A. 2006;103:16936-41.

(9.) Lee YJ, Shin JY, Song MS, Lee YM, Choi JG, Lee EK, et al. Continuing evolution of H9 influenza viruses in Korean poultry. Virology. 2007;359:313-23.

(10.) Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res. 1997;25:4876-82.

(11.) Perriere G, Gouy M. WWW-query: an on-line retrieval system for biological sequence banks. Biochimie. 1996;78:364-9.

(12.) Seo SH, Hoffmann E, Webster RG. Lethal H5NI influenza viruses escape host anti-viral cytokine responses. Nat Med. 2002;8:950-4.

(13.) Hatta M, Gao R Halfmann P, Kawaoka Y. Molecular basis for high virulence of Hong Kong H5N1 influenza A viruses. Science. 2001;293:1840-2.

Address for correspondence: Chul-Joong Kim, College of Veterinary Medicine, Chungnam National University, 220 Gung-Dong, Yuseoung Gu, Daejeon 305-764, South Korea; email:

Youn-Jeong Lee, * (1) Young-Ki Choi, ([dagger]) (1) Yong-Joo Kim, * Min-Suk Song, ([dagger]) Ok-Mi Jeong, * Eun-Kyoung Lee, * Woo-Jin Jeon, * Wooseog Jeong, * Seong-Joon Joh, * Kang-seuk Choi, * Moon Her, * Min-Chul Kim, * Aeran Kim, * Min-Jeong Kim, * Eun ho Lee, ([dagger]) Tak-Gue Oh, ([dagger]) Ho-Jin Moon, ([double dagger]) Dae-Won Yoo, ([double dagger]) Jae-Hong Kim, ([section]) Moon-Hee Sung, ([paragraph]) Haryoung Poo, # Jun-Hun Kwon, * and Chul-Joong Kim ([double dagger])

* Ministry of Agriculture and Forestry, Anyang, South Korea; ([dagger]) College of Medicine and Medical Research Institute of Chungbuk National University, Cheongju, South Korea; ([double dagger]) College of Veterinary Medicine of Chungnam National University, Daejeon, South Korea; ([section]) College of Veterinary Medicine of Seoul National University, Seoul, South Korea; ([paragraph]) Bioleaders Corporation, Daejeon, South Korea; and #Korea Research Institute of Bioscience and Biotechnology, Daejeon, South Korea

(1) These authors contributed equally to this work.

Dr Youn-Jeong Lee is a senior researcher at NVRQS. Her current research focuses on avian influenza viruses.
Table. History of highly pathogenic avian influenza (H5N1) suspected
cases, South Korea, 2006-2007

 Outbreak Source/
Date reported no. breed * Age, wk Region

2006 Nov 22 1 Chicken/BB 45 Jeollabuk
2006 Nov 27 2 Chicken/BB 40 Jeollabuk
2006 Dec 10 3 Quail 16-28 Jeollabuk
2006 Dec 21 4 Duck/B 30 Chungcheongnam
2007 Jan 20 5 Chicken/L 32 Chungcheongnam
2006 Dec 21 Environment Chungcheongnam
2006 Dec 21 Environment Chungcheongbuk

Date reported animals/farm Clinical signs Isolate

2006 Nov 22 13,000 Depression, A/chicken/IS/2006
 death A/chicken/IS2/2006
2006 Nov 27 12,000 Depression, A/chicken/IS3/2006
2006 Dec 10 290,000 Death A/quail/KJ4/2006
2006 Dec 21 9,000 Decrease in A/duck/Asan5/2006
 egg production A/duck/Asan6/2006
2007 Jan 20 30,000 Depression, A/chicken/CA7/2007
2006 Dec 21 A/environment/Korea/
2006 Dec 21 A/environment/Korea/

* BB, broiler breeder; B, breeder; L, layer.
COPYRIGHT 2008 U.S. National Center for Infectious Diseases
No portion of this article can be reproduced without the express written permission from the copyright holder.
Copyright 2008 Gale, Cengage Learning. All rights reserved.

Article Details
Printer friendly Cite/link Email Feedback
Title Annotation:DISPATCHES
Author:Lee, Youn-Jeong; Choi, Young-Ki; Kim, Yong-Joo; Song, Min-Suk; Jeong, Ok-Mi; Lee, Eun-Kyoung; Jeon,
Publication:Emerging Infectious Diseases
Geographic Code:9CHIN
Date:Mar 1, 2008
Previous Article:Hemagglutinating encephalomyelitis coronavirus infection in pigs, Argentina.
Next Article:Mutations in influenza A virus (H5N1)and possible limited spread, Turkey, 2006.

Related Articles
The root of bird flu?
Risk for infection with highly pathogenic influenza a virus (H5N1) in chickens, Hong Kong, 2002.
Genetic characterization of HPAI (H5N1) viruses from poultry and wild vultures, Burkina Faso.
Possible avian influenza (H5N1) from migratory bird, Egypt.
Experimental infection of swans and geese with highly pathogenic avian influenza virus (H5N1) of Asian lineage.
Avian influenza virus (H5N1) replication in feathers of domestic waterfowl.
Low frequency of infection with avian influenza virus (H5N1) among poultry farmers, Thailand, 2004.
Reassortant avian influenza virus (H5N1) in poultry, Nigeria, 2007.
Highly pathogenic avian influenza virus (H5N1) isolated from whooper swans, Japan.

Terms of use | Copyright © 2017 Farlex, Inc. | Feedback | For webmasters