• 한국어병학회지
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• 제34권1호
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• pp.9-15
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• 2021

Rock bream (Oplegnathus fasciatus) is a highly valued aquaculture species in Korea. However, the aquaculture industry suffers huge economic losses due to rock bream iridovirus (RBIV) infection in summer. The objective of this study was to determine genetic diversity and relationships of DNAs isolated from two groups of rock bream after RBIV infection using five microsatellite (MS) markers. The first group of fish died early and the second group of fish died later after RBIV infection. In this experiment, 90 fish (5.1±1.0 cm and 4.1±1.3 g) were injected with 50 μl of RBIV (10⁴ TCID⁵⁰/ml) and maintained at 26℃ for 15 days. Genomic DNAs were extracted from fins of 20 fish that died earlier or later after RBIV infection. These DNAs were subjected to genotyping using five MS markers (CA-03, CA3-05, CA3-06, CA-10, and CA3-36). Of these markers, CA3-05 (early death group), CA3-06 (late death group), and CA3-36 (both early and late death groups) showed different alleles distribution rates. In-depth studies are needed to provide valuable information for selecting RBIV-resistant fish. In conclusion, microsatellite marker distribution pattern differences between early- and late- death groups of rock bream after RBIV infection showing different RBIV susceptibilities were determined using MS markers and genotyping. Results of this study suggest that MS markers could be used to facilitate the selection of RBIV resistant rock bream.

• 수산해양교육연구
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• 제27권4호
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• pp.1184-1193
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• 2015

Monitoring for megalocytivirus infection was conducted for ten months from March to December in 2013 in 15 aquatic farms culturing, red sea bream, rock bream, rock fish and black sea bream around Tongyoung coastal area in Korea, to assess spatial and temporal variability of detection prevalence, and to explore possible links with seawater temperature. In nested-PCR targeted major capsid protein (MCP) gene, asymptomatic megalocytivirus infection was detected in the externally healthy farmed fish with a significant prevalence in range from 0 to 58.3% for ten months. Higher prevalence of megalocytivirus (46.7% - 57.1%) was observed in high water temperature season from September to November than that in other months with lower prevalence of 0.0% to 20.0%. Even though an acute infection of megalocytivirus was occurred in rock bream (positive in the first PCR) with high mortality in one of fifteen farms, there was no expansion or transmission of the disease to the rock fish and red sea bream culturing in net cage just proximal to the rock bream cage in which disease outbreaked. Nucleotide sequence analysis of the cloned MCP gene isolated asymptomatically infected rock fish revealed that the megalocytivirus in this study was clustered together with the rock bream iridovirus (RBIV) under the subgroup II of the genus megalocytivirus (Iridoviridae), which is known to be the major megalocytivirus strain in Korea. The typical histopathological signs were not found in the spleen of rock fish asymptomatically infected by megalocytivirus. Experimental infection of rock bream with the spleen homogenate of the rock fish infected asymptomatically did not induce any mortality unlike the homogenate of infected rock bream with hih mortlity. However, these results may suggest that the asymptomatic infection of megalocytivirus in other fish species can be a potential risk threatening aquaculture industries as a transmission factor of megalocytivirus to susceptible fish species, especially rock bream.

• 한국어병학회지
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• 제32권1호
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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.

• 한국어병학회지
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• 제36권2호
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• pp.191-211
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• 2023

Rock bream iridovirus (RBIV) causes high mortality and economic losses in rock bream (Oplegnathus fasciatus) aquaculture industry in Korea. Although, the immune responses of rock bream under RBIV infection have been studied, there is not much information at the different stages of infection (initial, middle and recovery). Gene expression profiling of rock bream under different RBIV infection stages was investigated using a microarray approaches. In total, 5699 and 6557 genes were significantly up- or down-regulated over 2-fold, respectively, upon RBIV infection. These genes were grouped into categories such as innate immune responses, adaptive immune responses, complements, lectin, antibacterial molecule, stress responses, DNA/RNA binding, energy metabolism, transport and cell cycle. Interestingly, hemoglobins (α and β) appears to be important during pathogenesis; it is highly up-regulated at the initial stage and is gradually decreased when the pathogen most likely multiplying and fish begin to die at the middle or later stage. Expression levels were re-elevated at the recovery stage of infection. Among up-regulated genes, interferon-related genes were found to be responsive in most stages of RBIV infection. Moreover, X-linked inhibitor of apoptosis (XIAP)-associated factor 1 (XAF1) expression was high, whereas expression of apoptosis-relate genes were low. In addition, stress responses were highly induced in the virus infection. The cDNA microarray data were validated using quantative real-time PCR. Our results provide novel inslights into the broad immune responses triggered by RBIV at different infection stages.

• 한국어병학회지
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• 제24권2호
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• pp.53-64
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• 2011

Megalocytivirus 감염 시 나타나는 임상적 증상의 정량적인 분석을 위하여, 돌돔에서 분리된 megalocytivirus IVS-1을 돌돔성어와 치어에 인위 감염시켰다. 그 결과 spleen index가 각각 $4.49{\pm}1.13$와 $4.85{\pm}1.06$로 나타 났으며, 이것은 정상배에 비해 3배 이상 증가한 값이었다. 폐사율은 돌돔성어와 치어에서 모두 100% 폐사한 반면 참돔치어는 30일이 지나도 60%의 폐사만 나타나 돌돔폐사율에 비해 낮은 폐사율을 나타냈다. 또한 IVS-1을 감염시킨 빈사상태의 참돔치어는 spleen index가 동일 크기의 돌돔치어 보다 낮은 $1.47{\pm}0.87$을 보였다. Real-time PCR을 이용하여 감염의 진행에 따른 바이러스 농도를 측정한 결과, 돌돔 치어와 성어의 감염 조직 내 바이러스 최고 농도는 각각 $2.03{\times}10^7$ copies/mg과 $2.40{\times}10^7$ copies/mg으로 비슷하였으나 돌돔성어에서 먼저 최고치에 도달하였다. 더구나 비장의 비정형비대세포의 수 역시 돌돔치어의 경우 지속적으로 증가한 반면 돌돔성어의 경우 최고치에 도달 후 오히려 감소히는 경향을 보였다. 본 연구에서는 megalocytivirus 감염 시 조직의 형태적 변화와 조직 내의 바이러스 농도와 같은 임상적인 증상을 정량적인 분석을 통해 나타내었고 이러한 정량적인 수치와 megalocytivirus 감염의 진행 정도와의 상관관계에 대해 조사하였다.

• 한국어병학회지
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• 제34권1호
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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.

• 한국어병학회지
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• 제27권2호
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• pp.85-89
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• 2014

Water temperature is a key environmental factor controlling the epizootics of viral diseases in fish. High water temperature is associated with the rapid spread of rock bream iridovirus (RBIV) disease and with high mortality of RBIV infected fish. Although protection of fish against iridoviral disease by active immunization has been reported, little information is available concerning whether fish survived from an epizootic of iridoviral disease can naturally acquire resistance against the viral disease. In the present study, we have demonstrated that juvenile rock bream, which survived from a natural epizootic of RBIV, acquired resistance against recurrence or reinfection of RBIV, and this resistance was established during the subsequent low water temperature period. Furthermore, the possible involvement of the adaptive humoral immune response in the resistance of the juvenile rock bream was suggested by in vivo neutralization experiment.

• 한국어병학회지
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• 제29권1호
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• pp.7-12
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• 2016

In a previous study on the parasites of cultured rock bream (Oplegnathus fasciatus), we reported the discovery of a new species, Ceratomyxa oplegnathus, obtained from the gallbladder. In the present study, we found another Ceratomyxa species, C. sparusaurati, also from the gallbladder of rock bream. The morphological and ultrastructural characteristics of C. sparusaurati were investigated using light and transmission electron microscopy (TEM).

• 한국발생생물학회지:발생과생식
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• 제21권4호
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• pp.371-378
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• 2017

Interferon-stimulated gene 15 (ISG15) is known to interfere with viral replication and infection by limiting the viral infection of cells. Interferon-stimulated gene 15 (ISG15) interferes with viral replication and infectivity by limiting viral infection in cells. It also plays an important role in the immune response. In this study, tissue-specific expression of ISG15 in healthy rock bream samples and spatial and temporal expression analysis of rock bream ISG15 (RbISG15) were performed following rock bream iridovirus (RSIV) infection. RbISG15 expression was significantly higher in the eye, gill, intestine, kidney, liver, muscle, spleen, and stomach, but low in the brain. There were particularly high levels of expression in the liver and muscle. RbISG15 expression was also examined in several tissues and at various times following RSIV infection. ISG15 expression increased within 3 h in the whole body and decreased at 24 h after infection. In addition, temporal expression of several tissues following RSIV infection showed a similar pattern in the muscle, kidney, and spleen, increasing at 3 h and decreasing at 72 h. These results suggest that ISG15 plays an important role in the immune response of rock bream. Overall, this study characterizes the response of RbISG15 following RSIV infection.

• 한국어병학회지
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• 제23권2호
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• pp.177-187
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• 2010

The Interferon stimulated gene 15 (ISG15) is strongly induced in many cell types by IFNs, viral infections, and double-stranded RNA (poly I:C). The ISG15 homologue cDNA was isolated from the rock bream LPS stimulated leukocyte cDNA library. The rock bream ISG15 homologue was found to consist of 833 bp encoding 157 amino acid residues. Compared with other known ISG15 peptide sequences, the most conserved regions of the rock bream ISG15 peptide were found to be the tandem ubiquitin-like domains and a C-terminal LRLRGG conjugating motif, characteristic of mammalian and non-mammalian ISG15 proteins. Phylogenetic analysis based on the deduced amino acid sequence revealed a homologous relationship between the ISG15 sequence of rock bream and that of Atlantic salmon, Atlantic cod, northern snake head, black rockfish and olive flounder. The expression of the rock bream ISG15 molecule was induced in the peripheral blood leukocytes (PBLs) from 1 to 24 h following poly I:C stimulation, with a peak at 3 h post-stimulation. The rock bream ISG15 gene was predominantly expressed in the PBLs, spleen and gill.