• Journal of fish pathology
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• v.34 no.2
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• pp.149-159
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• 2021

Genus Megalocytivirus cause red sea bream iridoviral disease (RSIVD) and scale drop disease (SDD). Based on the phylogeny of the major capsid protein (MCP) and adenosine triphosphatase (ATPase) genes, megalocytiviruses except for SDD virus (SDDV) could be three different genotypes, red sea bream iridovirus (RSIV), infectious spleen and kidney necrosis (ISKNV), and turbot reddish body iridovirus (TRBIV). In this study, primary cells derived from the caudal fin of rock bream (Oplegnathus fasciatus) grew at 25℃ in Leibovitz's medium supplemented with 10% (v/v) fetal bovine serum and primocin (100 ㎍/mL). Rock bream fin (RBF) cells exhibited susceptibility to infections by different genotypes of megalocytiviruses (RSIV, ISKNV and TRBIV) with the appearance of cytopathic effects with an increase in the viral genome copy number. Furthermore, compared to grunt fin (GF) cells, even though 10 times lower number of RSIV genome copies were inoculated in RBF cells, viral genome copy number produced on RBF cells were 44 times higher than that of GF cells at 7 d post-inoculation. As the isolated RBF cells are sensitive to different genotypes of megalocytiviruses (RSIV, ISKNV and TRBIV), they can be used for future studies regarding in vitro viral infection and subsequent diagnosis.

• Journal of fish pathology
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• v.20 no.3
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• pp.211-220
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• 2007

Red sea bream iridovirus disease (RSIVD) cause massive economic losses in marine aquaculture industry in Korea. The causative agent of this disease (RSIV) infects a wide range of fish species. The aims of this study were to monitor RSIV in wild marine fishes and to give critical information for controling the disease through prophylactic methods. Prevalence of the viral disease, geological distribution and reservoir of the virus were investigated using wild marine fishes captured in southern coast and east china sea for two years. (Polymerase Chain Reaction) PCR results showed that RSIV were detected in 39 (24.3%) out of 160 fish. MCP gene sequences of viral strains isolated in this study were closely related to that of a reference strain, red seabream-K, belonging to Megalocytivirus subgroup Ⅲ. The results suggest that some of wild marine fishes are RSIV carriers and may spread the pathogen directly to fish farmed in coastal area.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.44 no.4
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• pp.352-358
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• 2011

The presence of ISKNV-like viruses in various freshwater ornamental fish species imported from Asia was confirmed by polymerase chain reaction(PCR) amplification of the ATPase(adenosine triphosphatase) gene. Interestingly, molecular analyses of the Open Reading Frame 25(ORF25) region of these isolates based on the ISKNV(Infectious spleen and kidney necrosis virus) genome revealed the presence of various repetitive sequences. ORF25 repeat sequence length had no effect on cumulative mortality of rock bream Oplegnathus fasciatus challenged with tissue homogenates of infected pearl gourami, Trichogaster leeri; silver gourami, Trichogaster microlepis; blue gourami, or Trichogaster trichopterus. All isolates induce cumulative mortalities after 12 days of infection, confirming that ORF25 polymorphism did not affect the pathogenicity of ornamental fish megalocytiviruses that cross infect rock bream, a seawater fish. Also, no statistically significant differences in spleen index or viral copy number in infected tissues was detected between isolates with varying ORF25 repeat sequence lengths. However, further studies are necessary to fully characterize the functional characteristics of these polymorphisms in megalocytivirus disease in ornamental fishes.

• Journal of fish pathology
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• v.34 no.1
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• pp.1-7
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• 2021

Red sea bream iridovirus (RSIV), belonging to the genus Megalocytivirus, is the predominant cause of mortality in marine fishes in Korea, including rock bream (Oplegnathus fasciatus). Rockfish (Sebastes schlegelii) are the host fish for RSIV, exhibiting no clinical signs or mortality. Cohabitation challenges, which mimicked natural transmission conditions, were performed to evaluate viral transmission between rock bream and rockfish, and to determine the pathogenicity and viral loads. In cohabitation challenge, artificially RSIV-infected rock bream were the viral donor, and healthy rockfish were the recipient. The results showed that although the donor rock bream had 95-100 % cumulative mortality (>10⁸ viral genome copies/mg of spleen 7-14 days after viral infection), the recipient rockfish did not die, even when the viral genome copies in the spleen were >10⁵ copies/mg. These results indicated asymptomatic infections. Notably, in a reverse-cohabitation challenge (artificially RSIV-infected rockfish as the viral donor and healthy rock bream as the recipient), RSIV horizontally infected from subclinical rockfish to rock bream (10⁷ viral genome copies/mg of spleen 21 days after cohabitation) with 10-20% cumulative mortality. These results suggest that an asymptomatic, infected rockfish can naturally transmit the RSIV without being sacrificed.

• Journal of Marine Life Science
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• v.3 no.2
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• pp.45-50
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• 2018

Rock bream iridovirus (RBIV) is a megalocytivirus widely infected in various fish species in Korea, causing symptoms of acute inflammation and enlargement of spleen. In our previous study, RBIV induced the initial upregulation but later down-regulation of proinflammatory cytokines and IFN1 gene expression. Signal transducers and activators of transcriptions (STAT) are transcription factors involved in the regulation of immune genes including IFNs. This study was conducted to analyse the expression of STAT2. The expressional study of STAT2 gene was performed in head kidney and spleen upon RBIV infection and immune stimulants like LPS or poly I:C in vitro. Consequently, STAT2 gene expression pattern was different in head kidney and spleen as it was significantly up-regulated by LPS from 4 h to 8 h but down-regulated at 24 h while up-regulated by poly I:C at 8 h in head kidney while, in spleen, STAT2 gene expression was down regulated by LPS but significantly up-regulated by poly I:C. Upon RBIV stimulation, STAT2 gene expression was significantly down-regulated by high dose RBIV at 4 h but up-regulated at 8 h and 24 h in head kidney. In spleen cells, it was up-regulated by medium dose RBIV at 4 h and by high dose RBIV at 4 h and 8 h but down regulated later then. In vivo, STAT2 gene expression was not significantly affected by RBIV infection while significant up-regulated by vaccination at day 7 post-vaccination, indicating STAT2 gene can be involved in adaptive immune response in rock bream.

• Journal of fish pathology
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• v.23 no.3
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• pp.335-342
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• 2010

In 2008, mass mortality was observed in paradise fish Macropodus opercularis which was imported from Indonesia. PCR of these fish found positive for megalocytivirus and Mycobacterium sp., while an unidentified virus was culture-isolated using CHSE-214 cells. In the present study, we investigated characterization of the unidentified virus and its pathogenicity to determine whether the virus was the causative agent of the mass mortality of paradise fish. The unidentified virus induced cytopathic effect (CPE) with syncytia in CHSE-214 and other fish cells, BF-2, GF, SSN-1, FSP and FFN. The virus was resistant against treatments with IUdR, chloroform, acidity at pH 3, basicity at pH 11 and high temperature at $56^{\circ}C$ for 3h. By electron microscopy, the viral particles were spherical having a double capsid structure with approximately 65 nm in external diameter. Viral genome was composed of at least 10-segmented RNA with sizes ranging from 0.7 kb to 3.6 kb. Based on these characters, this virus can be classified into family Reoviridae. This reovirus did not cause any mortality in an artificial experiment conducted by injecting the virus to paradise fish. This indicates that the reovirus is not only responsible for the mass mortality of paradise fish in 2008.

• Journal of fish pathology
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• v.32 no.1
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• pp.1-8
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• 2019

Rock bream iridovirus (RBIV) is a member of the Megalocytivirus genus that causes severe mortality to rock bream (Oplegnathus fasciatus) with characteristic clinical signs of spleen enlargement. In this study, we assessed spleen size and RBIV copy number patterns in RBIV-infected rock bream to determine lethal and safe levels of virus copy number/spleen index that may define disease progress. We found that rock bream infected with RBIV ($1.1{\times}10^7virus\;copy\;number/100{\mu}l$) and held at 29, 26, 23 or $20^{\circ}C$ exhibited significantly higher levels of spleen size compared to $17^{\circ}C$. In dead condition (100% mortality at $20{\sim}29^{\circ}C$), the spleen index ($spleen\;weight/fish\;weight{\times}100$) and virus copy number were 3.00~5.38 and $10^6{\sim}10^8/{\mu}l$, respectively. Conversely, in survived condition (0% mortality at $17^{\circ}C$), spleen index and virus copy number was as low as not-infected control ($0.34{\sim}1.22/10^0{\sim}10^1/{\mu}l$, respectively). These findings suggest that spleen index can be an indicator of disease severity of RBIV disease.

• Fisheries and Aquatic Sciences
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• v.24 no.11
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• pp.351-359
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• 2021

The red sea bream iridovirus (RSIV) belonging to genus Megalocytivirus is responsible for red sea bream iridoviral disease (RSIVD) in marine and freshwater fishes. Although several diagnostic assays for RSIV have been developed, diagnostic sensitivity (DSe) and specificity (DSp) of real-time polymerase chain reaction (PCR) assays are not yet evaluated. In this study, we developed a TaqMan probe-based real-time PCR method and evaluated its DSe and DSp. To detect RSIV, the probe and primers were designed based on consensus sequences of the major capsid protein (MCP) genes from megalocytiviruses including RSIV, infectious spleen and kidney necrosis virus (ISKNV), and turbot reddish body iridovirus (TRBIV). The probe and primers were shown to be specific for RSIV, ISKNV, and TRBIV-types megalocytiviruses. A 95% limit of detection (LOD_95%) was determined to be 5.3 viral genome copies/µL of plasmid DNA containing the MCP gene from RSIV. The DSe and DSp of the developed real-time PCR assay for field samples (n = 112) were compared with those of conventional PCR assays and found to be 100% and 95.2%, respectively. The quantitative results for SYBR Green and TaqMan probe-based real-time PCR were not significantly different. The TaqMan probe-based real-time PCR assay for RSIV may be used as an appropriate diagnostic tool for qualitative and quantitative analysis.