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DETECTION AND ISOLATION OF HIGHLY PATHOGENIC H5N1 AVIAN INFLUENZA A VIRUSES FROM BLOW FLIES COLLECTED IN THE VICINITY OF AN INFECTED POULTRY FARM IN KYOTO, JAPAN, 2004

During the outbreak of highly pathogenic avian influenza that occurred in Tamba Town, Kyoto Prefecture in 2004, a total of 926 flies were collected from six sites within a radius of 2.3 km from the poultry farm. The H5 influenza A virus genes were detected from the intestinal organs, crop, and gut of the two blow fly species, Calliphora nigribarbis and Aldrichina grahami, by reverse transcription-polymerase chain reaction for the matrix protein (M) and hemagglutinin (HA) genes. The HA gene encoding multiple basic amino acids at the HA cleavage site indicated that this virus is a highly pathogenic strain. Based on the full-length sequences of the M, HA, and neuraminidase (NA) segments of virus isolates through embryonated chicken eggs, the virus from C. nigribarbis (A/blow fly/Kyoto/93/2004) was characterized as H5N1 subtype influenza A virus and shown to have > 99.9% identities in all three RNA segments to a strain from chickens (A/chicken/Kyoto/3/2004) and crows (A/crows/Kyoto/53/2004) derived during this outbreak period in Kyoto in 2004. Our results suggest it is possible that blow flies could become a mechanical transmitter of H5N1 influenza virus.

Received August 29, 2005. Accepted for publication April 25, 2006.

Acknowledgments: The authors thank Drs. Masaji Mase and Shigeo Yamaguchi of the National Institute of Animal Health, and Masaji Obuchi and Masato Tashiro of the Department of Virology III of NIID for facilitating this research. The authors thank Dr. Lisa Filippi of Hoffstra University for correcting the English of the original manuscript. We also thank the staff of Department of Medical Entomology of National Institute of Infectious Diseases for providing fly specimens from Tamba Town of Kyoto in 2004.

Financial support: This research was supported by Grant-in-Aid for Scientific Research from the Ministry of Health, Labor and Welfare of Japan (H17-Shinko-Ippan-017) and for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology, the Japanese Government (17659124).

^* Address correspondence to Kyoko Sawabe, Department of Medical Entomology, National Institute of Infectious Diseases, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo 185-0023, Japan. E-mail: sawabe{at}nih.go.jp

Authors’ addresses: K. Sawabe, K. Hoshino, H. Isawa, T. Sasaki, T. Hayashi, Y. Tsuda, H. Kurahashi, and M. Kobayashi, Department of Medical Entomology, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo 185-0023, Japan. K. Tanaba-yashi, A. Hotta, and A. Yamada, Department of Veterinary Science, National Institute of Infectious Diseases, Tokyo, Japan. T. Saito, Department of Virology III, National Institute of Infectious Diseases, Musashi Murayama-city, Gakuen 4-7-1, Tokyo, Japan. Present address of T. Saito: Department of Infectious Diseases, National Institute of Animal Health, Kannondai 3-1-15, Tsukuba, Ibaraki 305-0856, Japan.

Reprint requests: Kyoko Sawabe, Department of Medical Entomology, National Institute of Infectious Diseases, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo 185-0023, Japan. Telephone: 81-3-5285-1111, Fax: 81-3-5285-1147, E-mail: sawabe{at}nih.go.jp.