• Journal of Aquaculture
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• v.17 no.1
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• pp.24-29
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• 2004

The high mortality of cultured turbot, Scophthalmus maximus was occurred from Gochang in June, 2003. Infect fish with iridovirus showed a lower feed intake and lethargic. These fish exhibited pale body color, extended abdominal and exophthalmus. Histopathological studies showed basophilic enlarged cells from kidney, spleen, gill, heart, muscle, stomach, intestine, liver and pancreas of these fish. In PCR amplification with red sea bream iridovirus (RSIV) specific primer set of a DNA fragment of 286 bp was obtained from infect turbot with the virus. The strain showed the high homologies with RSIV, LBIV (largemouth bass iridovirus), GSIV (giant seaperch iridovirus) and SBIV (sea bass iridovirus).

• Journal of fish pathology
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• v.21 no.3
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• pp.271-278
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• 2008

The epidemiological study was performed to survey the prevalence of 5 bacterial, 6 viral and 5 parasitic disease of cultured olive flounder, Paralichthys olivaceus in Pohang, Ulsan∙Gijang, Keoje and Wando area of Korea from February to December, 2007. The disease frequency of size groups were as follows; Among the 1,218 fish samples, less than 10 cm (67.6%), 11~20 cm (70.2%), 21~30 cm (73.4%), 31~40 cm (77.6%), above 41 cm (88.2%). The highest detection rates of pathogens were recorded in samples from August and Ulsan. The detection rates of parasites, bacteria or viruses were 43.2%, 38.8% and 24.4%, respectively. The distribution of specific diseases showed that detection rates of diseases occurring the most frequently during the study period were Trichodina (33.1%), viral nervous necrosis virus (16.0%), Vibrio (9.7%), Scutica (10.1%), Ichthyobodo (5.7%).

• Journal of fish pathology
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• v.36 no.2
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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.

• Journal of fish pathology
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• v.23 no.1
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• pp.27-35
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• 2010

Pathogens on the cultured 579 rockfish, Sebastes schlegeli in the marine cage farms from Geoje and Tongyeong of the Southern sea were investigated from 2006 to 2008. The pathogens were detected throughout the year at 46.0~90.0% for 3 years and the detection rate was low with an average 58.1% in May and high with an average 81.5% in October. Bacteria only, bacteria-parasite mix and virus only were found in October and November as well as parasite only, whereas infection of parasite only was dominant in May when the temperature increased and in August when the temperature peaked. Of rockfish, Microcotyle sp. and Caligus sp. were dominant for parasitic disease, and Vibrio sp. and Streptococcus sp. were dominant bacteria. For virus, RSIV and VNNV were detected as dominant organisms. While no virus was detected in 2006, VNNV, VHSV and RSIV were detected in 2007 due to $1.5\sim2.0^{\circ}C$ higher temperature than 2006 in the summer season. For total prevalence by rockfish sizes, the highest was found at 50.0~87.1% in 11~15cm sizes and 50% was found in 30 cm size. Parasite showed a similar trend of 50.0~79.6% as the total prevalence. Prevalence for bacteria varied from 1.6% (for 10 cm group) to 23.8% (for 26 cm group) and higher virus prevalence of 21.5% was found from below 25 cm group.

• Journal of fish pathology
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• v.35 no.2
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• pp.167-176
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• 2022

We injected infectious myonecrosis virus (IMNV) to pacific whiteleg shrimp, Litopenaeus vannamei, and observed closely with using light microscope and transmission electron microscope (TEM) for 4-8 days post infection (dpi). As clinical signs, abdominal bodies had mild opaque muscles at 5 dpi. And the mortality was shown at 6 dpi. At 8 dpi, most injected shrimps had severe opaque muscles and humped back that cause of movement disorder. As results of histopathological examinations, local parts of abdominal body muscle had muscle fiber hyalinization, muscle fiber atrophy, rounded muscle fibers, myofibrillar hypertrophy in size, a decrease in number of myofibrils and phagocytosis from the sarcolemmas by multiple hemocytes at 4 dpi. Especially, myofibrillar hypertrophy appeared at the whole or random part of single muscle fiber not in specific locations like the center or edge of muscle fiber. At 6-7 dpi, multiple muscle necrosis, muscle fiber segmentation, myofibril lysis ap- peared and a few hemocytes were infiltrated at lesions. At 8 dpi, extensive muscle necrosis, multiple myofibril lysis and muscle fiber atrophy were shown, and very few hemocytes were infiltrated. In early stage of infection, local viral myositis with zenker's degeneration were shown. These lesions appeared multiply after the early stage. In late stage of infection, extensive coagulative muscle necrosis appeared with few of inflammatory response such as hemocytes infiltration. The lack of hemocytes infiltration response at the late stage might be disadvantage for Litopenaeus vannamei to defense against IMNV and to recover, because hematocytes (granulocyte, semi-granulocyte) eliminate pathogen and damaged tissues from infection sites and help recover. As results of the TEM observation, IMNVs that had nonenveloped icosahedral capsid which was 30-40 nm diameter were in myofibril and beside tubules of sarcoplasmic reticulum and moved to the certain direction. The micro-tears and micro-trau- mas in myofibrils caused muscle fiber necrosis. And semi-granulocytes engulfed IMNVs to eliminate virus.

• Journal of fish pathology
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• v.35 no.1
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• pp.27-40
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• 2022

Yellow head virus (YHV), Infectious hypodermal and hematopoietic necrosis (IHHNV), Taura syndrome virus (TSV), and Infectious myositis virus (IMNV) cause serious mortality to Penaeidae shrimp in the aquaculture. In this study, YHV, IHHNV, TSV, and IMNV were surveyed from imported frozen shrimps between 2019 and 2020 via molecular diagnostic assay. Among 10 shrimp groups, YHV (n=1) and IHHNV (n=4) were detected by RT-PCR and PCR, respectively. From the phylogenetic analysis based on the partial ORF 1b region of YHV, YHV was classified into YHV genotype 3 (YHV3). And IHHNVs (n=2) detected from Litopenaeus vannamei belong to infectious IHHNV type 2. Although IHHNVs (n=2) identified from Penaeus monodon showed PCR positive results (MG 831F/R primer set), the sequences of ORF 2 and 3 were not amplified, suggesting that those samples might possess type A IHHNV related sequence of P. monodon. Furthermore, in the challenge test, even though PCR-detected isolates (YHV3/type A IHHNV related sequence or infectious IHHNV type 2) were not induced mortality to L. vannamei, viral genes were amplified suggesting that the viruses in the frozen shrimp could be non-pathogenic particles which are not enough to induce mortality.

• Journal of fish pathology
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• v.35 no.2
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• pp.225-230
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• 2022

Viral hemorrhagic septicemia (VHS) is one of the most serious viral diseases affecting farmed olive flounder (Paralichthys olivaceus) in Asian countries. VHS, caused by viral hemorrhagic septicemia virus (VHSV), occurs in over 80 different cultured and wild fish species worldwide. Our previous study demonstrated that VHSV infection can be restricted by adjusting the water temperature to over 17℃ from the host optima. We confirmed that the effective VHSV immersion vaccine treatment was a tissue culture infection dose (TCID) of 10^5.5 TCID₅₀/mL at 17℃. However, the safety of live VHSV immersion vaccines remains unclear. The objectives of this study were to 1) demonstrate the safety of the live VHSV immersion vaccine under co-habitant conditions and 2) estimate the pathogenicity of VHSV in live VHSV-vaccinated flounder at 10℃. No mortality was observed in olive flounder treated with the live VHSV immersion vaccine, and the vaccinated flounder challenged with VHSV did not transfer VHSV to naïve fish at 10℃ through cohabitation. VHSV titration was below the detection limit (< 1.3 log TCID₅₀/mL) in live VHSV immersion vaccine-treated flounder challenged with VHSV at 10℃. This study demonstrated that flounder treated with the live VHSV immersion vaccine were resistant to VHSV infection, and the live vaccine was also safe for naïve fish even at a water temperature known to be VHS infectious.

• Journal of fish pathology
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• v.36 no.2
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• pp.239-250
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• 2023

Disease monitoring was conducted to investigate the recent disease occurrence in Japanese eels (Anguilla japonica). Between May 2021 and March 2022, an investigation was conducted on eels from seven farms experiencing mortality. JEECV (Japanese eel endothelial cells-infecting virus) was detected in all examined farms, each exhibiting co-infections with 1 or 2 bacteria, including Edwardsiella anguillarum, E. piscisida, Aeromonas sp., Citrobacter freundii, Lactococcus garviae, or Vibrio sp. From March 2022 to October 2023, monthly periodic inspections were carried out at a farm in Yeonggwang, Jeollanam-do, for a total of 22 times. JEECV was detected in 10 out of 22 times, even when mortality was not recorded. Bacteria such as E. anguillarum, C. freundii, Aeromonas sp., and Vibrio sp. were isolated, but consistent clinical signs of liver abscess and hemorrhagic lesions were only recognized in fish infected with E. anguillarum. Other bacteria were often isolated from apparently healthy fish. In conclusion, mortality in eel farms frequently occurs due to co-infections of JEECV with bacteria rather than JEECV alone. Therefore, to reduce eel mortality, it is crucial to decrease co-infections, with a particular emphasis of JEECV and E. anguillarum.

• Journal of Aquaculture
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• v.15 no.1
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• pp.7-13
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• 2002

Since 1993, the White Spot Syndrome Virus (WSSV) disease occurred in China among cultured shrimps resulting in mass mortality. Epizootiological surveys undertaken during the outbreak period of 1993-1994 indicated that all stages of Penaeus chinensis, P. japonicus and P. monodon were infected. Consequent to the transport of contaminated shrimp seedlings and seawater, the disease spread all over the farms of China. The disease was more rapidly transmitted at temperatures above $25^{\circ}C$. Challenge experiments showed the causative agent was highly virulent. White spots appeared on the carapace of both span-taneous and experimentally infected shrimps. Moribund shrimps contained turbid hemolymph, hypertrophied Iymphoid organ and a necrotic mid-gut gland. Electron microscopy showed the presence of viral particles in the gills, stomach, lymphoid organ, and epidermal tissue of the infected shrimp. The visions were slightly ovoid with an envelope and averaged 350 $\times$ 150 nm; nucleocapsids measured 375 $\times$ 157 nm. With discontinuous sucrose gradient of 35, 50 and 60% (w/v), the virus was separated from hemolymph of the infected shrimp. The estimated molecular weight of genomic DNA was 237 Kb with EcoR I, 247 Kb with Hind III and 241kb with Pst I. A total of 9 hybridoma colones secreting monoclonal antibodies (MAbs) were produced from mouse myeloma and spleen cells immunized with WSSV. The immunofluorescence assay of gill tissue showed that the MAbs reacted with diseased but not with healthy shrimp. The MAbs belonged to IgGl, IgG2b subclass and IgM class, all with kappa light Immune-electron-microscopy with colloidal gold marker showed the presence of 5 MAbs epitopes on the envelope and one on the capsid of the virus. Baculoviral mid-gut gland necrosis showed the specificity of the MAbs produced. For diagnosis 5 different methods were selected. Using Kimura primers for PCR, or MAbs for immunoblot, ELISA or FAT method, in situ hybridization was carried out to show the gene. All these methods detected WSSV in the organ samples of the diseased shrimp but not in healthy one.

• Genomics & Informatics
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• v.20 no.3
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• pp.33.1-33.17
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• 2022

Nervous necrosis virus (NNV) is a deadly infectious disease that affects several fish species. It has been found that the NNV utilizes grouper heat shock cognate protein 70 (GHSC70) to enter the host cell. Thus, blocking the virus entry by targeting the responsible protein can protect the fishes from disease. The main objective of the study was to evaluate the inhibitory potentiality of 70 compounds of Azadirachta indica (Neem plant) which has been reported to show potential antiviral activity against various pathogens, but activity against the NNV has not yet been reported. The binding affinity of 70 compounds was calculated against the GHSC70 with the docking and molecular dynamics (MD) simulation approaches. Both the docking and MD methods predict 4 (PubChem CID: 14492795, 10134, 5280863, and 11119228) inhibitory compounds that bind strongly with the GHSC70 protein with a binding affinity of -9.7, -9.5, -9.1, and -9.0 kcal/mol, respectively. Also, the ADMET (absorption, distribution, metabolism, excretion, and toxicity) properties of the compounds confirmed the drug-likeness properties. As a result of the investigation, it may be inferred that Neem plant compounds may act as significant inhibitors of viral entry into the host cell. More in-vitro testing is needed to establish their effectiveness.