[논문]Is Koi Herpesvirus (KHV) Related to the Mass Mortality Occurring among Cultured Carp, Cyprinus carpio, in Korea?

Kim, Wi-Sik; Jung, Sung-Ju; Kim, Du-Woon; Kim, Seok-Ryel; Kim, Jeong-Ho; Oh, Myung-Joo

doi:10.5657/fas.2010.13.1.079

Is Koi Herpesvirus (KHV) Related to the Mass Mortality Occurring among Cultured Carp, Cyprinus carpio, in Korea? 원문보기

Fisheries and aquatic sciences, v.13 no.1, 2010년, pp.79 - 83

Kim, Wi-Sik (Department of Aqualife Medicine, College of Fisheries and Ocean Science, Chonnam National University) , Jung, Sung-Ju (Department of Aqualife Medicine, College of Fisheries and Ocean Science, Chonnam National University) , Kim, Du-Woon (Division of Food Science and Technology and Functional Food Research Center, Chonnam National University) , Kim, Seok-Ryel (Pathology Division, National Fisheries Research and Development Institute) , Kim, Jeong-Ho (Faculty of Marine Bioscience and Technology, Gangneung-Wonju National University) , Oh, Myung-Joo (Department of Aqualife Medicine, College of Fisheries and Ocean Science, Chonnam National University)

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Since 1998, a new viral disease with high mortality has been consistently recorded in Korea in cultured carp, Cyprinus carpio. In this study, we investigated an epizootic of the disease that caused high mortality rates in carp obtained from 11 farms in Korea between 1999 and 2007. Assessment of koi herpesvirus (KHV) levels in diseased carp was carried out to determine if this virus was the etiologic agent of disease in this instance. High mortality rates in carp were recorded mainly in the spring and autumn at water temperatures between $19^{\circ}C$ and $24^{\circ}C$. Diseased fish typically showed surface discoloration, with a thick opaque mucus covering the body and gills. Protozoan parasites and bacteria were recovered from 7/29 (24%) and 2/26 (8%) of fish, respectively. Evidence of viral infection was marked; cytopathic effects (CPEs), characterized by cell rounding and an extended cytoplasm in fathead minnow (FHM) cells, were detected in 40/41 fish (98%). A high mortality rate (80%) resulted when supernatants of cell cultures showing CPEs were applied to previously healthy fish. KHV was detected by polymerase chain reaction in 6/41 fish (15%), but was not detected in supernatants obtained from cell cultures showing CPEs. These results suggest that KHV may not be the etiologic agent of the high mortality occurring among cultured carp in Korea; therefore, some other-as yet unidentified-infective agent must be responsible.

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Samples from the JS99-2 and US02 farms were all positive, while 2/4 fish (50%) from the HD00 farm were KHV-positive. Furthermore, sequencing of the PCR products amplified from the JS99, HD00, and US02 samples had 100% homology with KHV In this study, we hypothesized that KHV was responsible for the mass mortality of carp in Korea. However, KHV was detected in only 6/41 fish (15%), suggesting that the majority of diseased fish were not infected with this virus.
Therefore, these data suggested that a new, and as yet unidentified, virus was causing mass morality in cultured carp in Korea. We hypothesized that the causative agent may be KHV due to the similarity of disease clinical signs, outbreak time, and outbreak temperature to those seen in outbreaks of KHV disease (KHVD). However, as yet no evidence exists in the published literature to support this hypothesis.

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2 mM of each dNTP, 1U Taq DNA polymerase, and template DNA. Amplification was performed using a GeneAmp 2400 thermal cycler (Perkin Elmer, USA) and 39 cycles (94℃ for 1 min, 64℃ for 1 min, and 72℃ for 1 min). PCR products were resolved on 1.
5% agarose gels and bands excised and purified using a QIAquick gel extraction kit (Qiagen, USA). Amplified products were subjected to nucleotide sequence analysis using an ABI PRISM dye terminator sequencing chemistry (Applied Biosystems, USA) with KHV9/5F and KHV9/5R primers, according to the manufacturer's instructions. The resulting sequences were assembled with Genetyx Win Ver.
, 2005). To investigate any possible relationship between the disease we report here and KHY PCR assays targeting this virus were performed using the 41 tissue homogenates and 13 supernatants obtained from cell cultures showing CPEs (Table 1). Six of 41 fish (15%) were KHV-positive.

참고문헌 (10)

Gilad O, Yun S, Andree KB, Adkison MA, Zlotkin A, Bercovier H, Eldar A and Hedrick RP. 2002. Initial characteristics of koi herpesvirus and development of a polymerase chain reaction assay to detect the virus in koi, Cyprinus carpio koi. Dis Aquat Org 48, 101-108.

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Haenen O, Way K, Bergmann SM and Ariel E. 2004. The emergence of koi herpesvirus and its significance to European aquaculture. Bull Eur Assoc Fish Pathol 24, 293-307.
Hedrick RP, Gilad O, Yun S, Spangenberg JV, Marty GD, Nordhausen RW, Kebus MJ, Bercovier H and Eldar A. 2000. A herpesvirus associated with mass mortality of juvenile and adult koi, a strain of common carp. J Aquat Anim Health 12, 44-57.

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Hutoran M, Ronen A, Perelberg A, Ilouze M, Dishon A, Bejerano I, Chen N and Kotler M. 2005. Description of an as yet unclassified DNA virus from diseased Cyprinus carpio species. J Virol 79, 1983-1991.

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Miyazaki T, Kuzuya Y, Yasumoto S, Yasuda M and Kobayashi T. 2008. Histopathological and ultra-structural features of koi herpesvirus (KHV)-infected carp Cyprinus carpio, and the morphology and morphogenesis of KHY. Dis Aquat Org 80, 1-11.

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Oh MJ, Jung SJ, Choi TJ, Kim HR, Rajendran KV, Kim YJ, Park MA and Chun SK. 2001. A viral disease occurring in cultured carp Cyprinus carpio in Korea. Fish Pathol 36, 147-151.

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Pokorova D, Vesely T, Piackova V, Reschova S and Hulova J. 2005. Current knowledge on koi her-pesvirus (KHV): a review. Vet Med-Czech 50, 139-147.
Sano M, Ito T, Kurita J, Yanai T, Watanabe N, Miwa Sand Iida T. 2004. First detection of koi herpesvirus in cultured common carp Cyprinus carpio in Japan. Fish Pathol 39, 165-167.

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Seo JW, Kim WS, Kim JO, Kim DW, Jung SJ and Oh MJ. 2010. Histopathology of unidentified viral disease of cultured carp, Cyprinus carpio. J Fish Pathol submitted.
Waltzek TB, Kelley GO, Stone SM, Way K, Hanson L, Fukuda H, Hirono I, Aoki T, Davison AD and Hedrick RP. 2005. Koi herpesvirus represents a third cyprinid herpesvirus (CyHV-3) in the family Herpesviridae. J Gen Virol 86, 1659-1667.

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Is Koi Herpesvirus (KHV) Related to the Mass Mortality Occurring among Cultured Carp, Cyprinus carpio, in Korea? 원문보기

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