DAO 105:193-202 (2013)  -  DOI: https://doi.org/10.3354/dao02627

Growth of cyprinid herpesvirus 2 (CyHV-2) in cell culture and experimental infection of goldfish Carassius auratus

Takafumi Ito1,*, Jun Kurita2,6, Akiyuki Ozaki3, Motohiko Sano4,7, Hideo Fukuda5, Mitsuru Ototake4,8 

1Tamaki Laboratory, Aquatic Animal Health Division, 224-1 Hiruta, Tamaki, Mie 519-0423, Japan
2Diagnosis and Training Center for Fish Diseases, 3Aquaculture Technology Division, 4Aquatic Animal Health Division, National Research Institute of Aquaculture, Fisheries Research Agency, 422-1 Nakatsuhamaura, Minami-Ise, Mie 516-0193, Japan
5Faculty of Marine Science, Tokyo University of Marine Science and Technology, 4-5-7 Konan, Minato, Tokyo 108-8477, Japan
6Present address: Headquarters, Fisheries Research Agency, 15F Queen’s Tower B, 2-3-3 Minato Mirai, Nishi, Yokohama, Kanagawa 220-6115, Japan 7Present address: Faculty of Marine Science, Tokyo University of Marine Science and Technology, 4-5-7 Konan, Minato, Tokyo 108-8477, Japan
8Present address:
Research Center for Aquatic Genomics, National Research Institute of Fisheries Science, Fisheries Research Agency, 2-12-4 Fukuura, Kanazawa, Yokohama, Kanagawa 236-8648, Japan

ABSTRACT: Herpesviral haematopoietic necrosis has caused great economic damage to goldfish Carassius auratus aquaculture in Japan. The existence of cyprinid herpesvirus 2 (CyHV-2), the causative agent, has also been reported from several other countries. To prevent spread to other areas, basic virological information such as viral kinetics in infected fish is essential. Experimental infection trials using reliably prepared CyHV-2 for defining viral kinetics are difficult to carry out because successful and sustainable propagation of this virus in cell culture has previously been limited. Here we describe a method for sustainable propagation of CyHV-2 in cell culture, and the results of fish infection experiments using the propagated virus. We found that goldfish fin (GFF) cells and standard Ryukin Takafumi (SRTF) cells established from goldfish fin can be used for continuous propagation of CyHV-2. Experimental infections using 2 varieties of goldfish, Ryukin and Edonishiki, were performed with the virus passaged 7 times in GFF cells. In transmission experiments with water temperature at 20°C, cumulative mortality was 30% in Ryukin infected by immersion, and 90 and 100% in Edonishiki and Ryukin intraperitoneally injected with the virus, respectively. In an experiment carried out at 25°C, 90% of Edonishiki challenged by immersion died. PCR detection of viral DNA from the organs of infected fish showed that systemic infection occurs and also that the kidney is a main viral multiplication site. Moreover, CyHV-2 was successfully re-isolated in GFF cells from the dead fish.


KEY WORDS: Herpesviral haematopoietic necrosis · Cyprinid herpesvirus 2 · Goldfish fin (GFF) cells · Experimental infection · Viral kinetics


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Cite this article as: Ito T, Kurita J, Ozaki A, Sano M, Fukuda H, Ototake M (2013) Growth of cyprinid herpesvirus 2 (CyHV-2) in cell culture and experimental infection of goldfish Carassius auratus. Dis Aquat Org 105:193-202. https://doi.org/10.3354/dao02627

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