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KMID : 1204320070230030257
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Laboratory Animal Research
2007 Volume.23 No. 3 p.257 ~ p.263
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Isolation of Duck Hepatitis Virus and Development of its Live Attenuated Vaccine by Duck Embryo Liver Cell Culture
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Park Su-Jin
Cho Kyoung-Oh
Kang Mun-Il
Lee Chai-Yong
Park Sang-Ik
Kim Ha-Hyun
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Abstract
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Duck hepatitis virus (DHV) is the causative agent of duck viral hepatitis, an acute and fatal disease of young duckling from two days to three weeks. The major pathologic change in DHV-infected duckling is hepatitis including necrotic foci and hemorrhagic spots. Although commercial vaccine against the type 1 DHV is developed and selling in Korea, duck viral hepatitis still occurred in Korea even in vaccinated farms. Therefore, the aim of this study is to isolate and identify currently circulating DHV and develop cell-culture adapted live vaccine. By serial duck embryo liver (DEL) cell passages, 6 DHVs were isolated form field cases showing characteristic opisthotonos and severe necrosis of hepatocytes. These viruses were confirmed as type I DHV by immunofluorescence test with chicken anti-DHV polyclonal antibody. In order to develop attenuated DHV vaccine, 60th passages of DHV in DEL cell cultures was carried out. The titration of virus was increased gradually from the 1th to 60th. After the 40th passage, the pathogenicity of virus for duckling was disappeared. All one-day-old ducklings vaccinated by oral or peritoneal administration with the DEL cell-adapted virus (DVH03-21 strain) were survived after challenge inoculation of a virulent DRL-62 strain at 7 days post-vaccination. The minimum immune dose of the DEL cell-adapted virus (DVH03-21 strain) was 1 × 103.0 TCID50/0.1 ㎖ in 1-day-old ducklings. After we inoculated 100 times (DVH03-21 strain) of the minimum immune dose in 1-day-old ducklings, the DEL cell-adapted virus (DVH03-21 strain) did not cause any clinical sings and pathogenic lesions. From these results, it concludes that currently occurred duck viral hepatitis is caused by type 1 DHV and cell-adapted live attenuated type 1 DHV vaccine promises highly safety and efficacy.
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KEYWORD
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Duck hepatitis virus, duck embryo liver cell, attenuation, vaccine
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