• Journal of fish pathology
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• v.36 no.2
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• pp.229-238
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• 2023

In this study, the correlation between clinical changes and RSIV genome copy number was investigated to determine the quantitative criteria for the characteristics of RSIV infection. The rock bream (Oplegnathus fasciatus) was intraperitoneally injected with three different doses (1.0×10¹, 1.0×10³ and 1.0×10⁵ viral genome copies/fish) as low, medium, and high doses, respectively. The clinical signs (spleen enlargement, death) observation and real-time PCR were conducted at 5, 10 and 14 days post-injection. During the experiment, spleen index as a quantitative indicator for spleen enlargement was continuously increased in the medium- (up to 2.26) and high-dose (up to 4.99) challenge groups, respectively. Notably, when the spleen index was over 1.5, 2.0, 2.5 and 3.0, a positive correlation was revealed with average viral genome copy numbers of 2.51, 3.37, 4.97 and 5.43×10⁷ viral genome copies/mg, respectively. Moreover, the threshold of spleen index over 1.5 was 1.0×10⁶ viral genome copies/mg, while the thresholds of spleen index over 2.0 and dead was 2.51×10⁷ viral genome copies/mg and the thresholds of spleen index over 2.5 and 3 was 3.98×10⁷ viral genome copies/mg. These findings suggest the possibility of quantitatively analyzing the characteristics and development process of RSIV infection.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.15 no.8
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• pp.5412-5418
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• 2014

The red seabream iridovirus (RSIV), which belongs to the iridoviridae, causes infectious fish diseases in many Asian countries, leading to considerable economic losses to the aquaculture industry. Using the yeast surface display (YSD) technique, a new experimental system was recently developed for the detection and identification of a variety of marine viruses. In this study, a coat protein gene of RSIV was synthesized based on the nucleotide sequence database and subcloned into the yeast expression vector, pCTCON2. The expression of viral coat proteins in the yeast strain, EBY100, was detected by flow cytometry and Western blot analysis. Finally, they were isolated from the yeast surface through a treatment with ${\beta}$-mercaptoethanol. The data suggests that the YSD system can be a useful method for acquiring coating proteins of marine viruses.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.47 no.3
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• pp.241-246
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• 2014

Viruses in the genus Megalocytivirus have been subdivided into four subgroups. Among these subgroups 2 and 4, represented by the red sea bream iridovirus (RBIV) and the olive flounder iridovirus (FLIV), respectively, are non-exotic. subgroups 1 and 3, represented by the red sea bream iridovirus (RSIV) and the infectious spleen and kidney necrosis virus (ISKNV), respectively, have not been detected in Korea and are known as exotic. Shellfish are filter-feeders, and can thus filter and accumulate Megalocytivirus in their digestive glands, allowing us to track viral contamination in surrounding aquatic environment. In this study, we developed a high-resolution melting (HRM) analysis to differentiate among subgroups of Megalocytivirus accumulated in shellfish, and confirmed the convenience and efficiency of this method. More than two subgroups of Megalocytivirus were found in the digestive gland of a single shellfish. We classified all Megalocytivirus viruses from shellfish in Korea into subgroups 2 and 4, although proportions of subgroups were different among regions. Compared to nucleotide sequencing analysis, HRM analysis is a simple and rapid method for differentiating of Megalocytivirus subgroups.

• Fisheries and Aquatic Sciences
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• v.26 no.2
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• pp.97-104
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• 2023

CD96 is a membrane-bound receptor discovered in humans in 1992 that is mainly present in natural killer cells and T cells derived from haematopoietic cells and performs immune functions. Based on the sequence of CD96 obtained from red seabream (Pagrus major), phylogenetic analysis with other species, infections of normal fish, Streptococcus iniae and red sea bream iridovirus (RSIV), and expression analysis was conducted using real-time polymerase chain reaction. Phylogenetic analysis showed the highest homology with Sparus aurata, and multiple sequence analysis confirmed the conservation of major domains between different fish species. Normal fish high expression results were confirmed in the head kidney, and spleen, which are the haematopoietic organs of the fish. High expression levels were confirmed in the gills, liver, spleen, and kidney on day three after RSIV infection. After S. iniae infection, high expression was confirmed in the gills and liver on day one, and high expression was confirmed in the spleen from 12 hours. These results show that PmCD96 functions as an immune gene in P. major and is considered a basic research case for CD96 in fish's hematopoietic organ immune system.

• Journal of fish pathology
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• v.23 no.1
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• pp.85-98
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• 2010

Ministry of Agriculture, Fisheries and Food (MAFF) and National Fisheries Research and Development Institute (NFRDI) have inspected the hatchery-reared seeds of 22 marine species and 11 freshwater species for aquatic animal diseases in stock enhancement program in 2009. Results showed that total 12 local selfgovernments have been restocking the sea with cultured juveniles. Gyeongsangnam-do, Jeollanam-do, Jejudo and Chungcheongnam-do have a preference for marine species seeds to freshwater species. On the contrary, freshwater species were released mostly in Gyeonggi-do, Jeollabuk-do and Chungcheongbuk-do. In the marine species group, abalone was the most abundant as 24.5%, and then sea cucumber (15.2%), olive flounder (11.5%), swimming crab (5.6%), black sea bream and rockfish (6.8%), rock bream (5.1%), black rockfish (4.6%) and scorpionfish (4.5%) were followed. Crucian carp was the most abundant as 19.4%, and then eel (17.0%), Korean bullhead (12.3%), melanian snail (12.0%), catfish (8.4%) were followed in the freshwater species group. The total number of inspection cases in this study were 1,080 and disqualification cases were 19 by detection of aquatic animals pathogens such as red sea bream iridovirus (RSIV), koi herpesvirus (KHV) or white spot syndrome virus (WSSV).

• Journal of fish pathology
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• v.24 no.2
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• pp.121-129
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• 2011

Aquatic animal raised in hatcheries play an important role in supplying seedling to stock enhancement and seed quality, especially, seed health is the key factor for survival in the field after release and for stocking effectiveness. We have inspected the hatchery-reared seeds of 33 marine species and 12 freshwater species for legally designated diseases in stock enhancement program in 2010. Results showed that abalone was the most abundant as 20.0% in the marine species group and then sea cucumber (15.6%), olive flounder (8.4%), rockfish (6.7%), black sea bream (6.3%) and swimming crab (6.1%) were followed. Crucian carp was the most abundant as 19.4%, and then eel (11.8%), Korean bullhead (10.9%), mandarin fish (10.8%), melanian snail (8.4%), catfish (7.7%) were followed in the freshwater species group. The total number of inspection cases for eight pathogens in this study were 2,105 and disqualification cases were 30 by detection of aquatic animals pathogens such as koi herpesvirus (KHV), red sea bream iridovirus (RSIV), white spot syndrome virus (WSSV) or viral haemorrhagic septicemia virus (VHSV).

• Journal of fish pathology
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• v.13 no.2
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• pp.121-127
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• 2000

From August to October 1998, over 60% mortality of cultured striped beakperch Oplegnathus fasciatus was occurred in net cages along the southern coast of Korea. Moribund fish showed some clinical signs of lethargic behavior, dark coloration or decoloration, severe gill anemia and enlargement of spleen. Also enlarged basophilic cells showing Feulgen -positive reaction were observed in the tissue section of spleen, kidney, liver and heart of the diseased fish. GF cells inoculated with spleen homogenate of diseased fish produced cytopathic effect of enlarged and rounded cells, therefore the causative virus was isolated from diseased fish. Striped beakperch fingerlings intraperitoneally inoculated with the causative virus ($10^4TCID_{50}$/0.1 ml) revealed symptoms similar to those of naturally infected fish and died from 7 to 14 days post injection. Transmission electron microscopy revealed that the causative virus was enveloped icosahedral particle with 120~130 nm in diameter. PCR products of the expected size (500 bp) were amplified with a primer set based on the ATPase gene of RSIV(red sea bream iridovirus) using template DNAs which were extracted from the spleen of diseased fish and GF cells inoculated with the causative virus. According to the analysis of nucleotide sequence of these PCR products, the sequence from ATPase cDNA gene of the causative virus showed 95% homology with that of RSIV. These results indicate that the mass mortality in the cultured striped beakperch was caused by the infection of iridovirus similar to RSIV.

• Journal of fish pathology
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• v.26 no.3
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• pp.163-171
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• 2013

Polymerase chain reaction (PCR) is the most rapid and widely used method to detect viral pathogens. However, this method does not provide histopathologic nature of the virus. In situ hybridization (ISH) with oligonucleotide probes is attractive because it is a rapid method for detection and identification of viral pathogens at sites of tissue infection. In order to understand the histopathologic characterictics of Red sea bream iridovirus (RSIV), viral-hemorrhagic septicemia (VHS) virus and viral nervous necrosis (VNN) virus to cultured olive flounder, we her applied ISH method to various kinds of olive flounder tissues with PCR-positive for these three viruses. We found that these viruses showed different tissue tropism and were detected from different cell types. Our results suggest that ISH is useful not only in rapid detection of viral pathogens but also in understanding the histopathologic characters of specific viral pathogens.

• Journal of fish pathology
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• v.22 no.1
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• pp.75-83
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• 2009

Disease surveillance was performed to monitor the prevalence of fish pathogens in wild marine fish caught in coastal offshore water from February to October in 2008. A total of 401 fish samples were collected at set net or fish market at landing port on the coast of Pohang, Geoje, Yeosu and Jeju. In this study, 17 kinds of fish pathogens were isolated from 152 fish samples. The detection rates of parasites, bacteria or viruses were 21.4%, 17.0% and 2.7%, respectively. The detected parasites were Scutica, Trichodina, Cryptocaryon, Dactylogyrus, Microcotyle, Benedenia, Bivagina, Heteraxin, Caligus, Epistylis and nematode. The dominant bacterial pathogens were Vibrio, Streptococcus, Photobacterium and Psuedomonas. Red sea bream iridovirus (RSIV) and lymphocystis disease virus (LDV) were detected in 6 species of fish virus examined in this study. The detection rates of fish pathogens from Scorpaenidae, Monacanthidae, Pleuronectidae, Sparidae and Carangidae investigated over 30 samples were 59.2%, 48.4%, 34.2%, 30.6% and18.2%, respectively.

• Journal of fish pathology
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• v.25 no.3
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• pp.263-270
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• 2012

This study surveyed for the prevalence of parasites, bacteria and viruses in four fish species, olive flounder (Paralichthys olivaceus), red sea bream (Pagrus major), black sea bream (Acathopagrus schlegeli) and black rockfish (Sebastes schlegeli) in 2010. The survey was aimed to compare the pathogens detected from wild and cultured fish for an epidemiological study. Anisakis sp. was predominantly detected from wild olive flounder and red sea bream (58.6% and 41.7% respectively), but not from the cultured fishes, suggesting anisakid infection is rare in cultured fish. The wild fish get in contact with the anisakids through their prey such as small fishes or crustaceans which carry the anisakids; whereas the cultured fish are fed with formulated feed, free of anisakids. Bacterial detection rates from the wild fishes examined in the study were lower than those of cultured fishes. Vibrio sp. dominated among detected bacterial population in cultured olive flounder (18%). Since vibriosis is known as a secondary infection caused by other stressful factors such as parasitic infections, handling and chemical treatment, it seems that cultured olive flounder are exposed to stressful environment. Viruses diagnosed in the study showed difference in distribution between wild and cultured fishes; hirame rhabdovirus (HRV) (0.1%) and lymphocystis disease virus (LCDV) (3.9%) were detected in the cultured olive flounder, but not in the wild fish, and marine birnavirus (MBV) (1.7%) and red sea bream iridovirus (RSIV) (3.2%) were detected from the wild and cultured red sea bream, respectively. From the survey conducted, it can be concluded that even though some pathogens (Trichodina sp., Microcotyle sp., etc.) are detected from both the wild and cultured fish, pathogens such as Anisakis sp., Vibrio sp. and LCDV showed difference in distribution in the wild and cultured host of same fish species and this can be attributed to their environmental condition and feeding.