• Journal of fish pathology
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• v.11 no.1
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• pp.61-67
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• 1998

For rapid detection of iridovirus infection, a PCR-based diagnostic method was developed. The genomic DNA from mortality-associated iridovirus was cloned into pUC19 vector. The nucleotide sequences of these clones were compared with sequences of other genes from EMBL/GenBank databank. Based on the nucleotide sequences, PCR primers were prepared and used for PCR. The DNA amplification did not occur from the normal fish cells. In contrast, DNA was amplified from the iridovirus-infected fish cells and purified iridovirus. These results suggest that mortality-associated iridovirus can be detected from virus-infected cells within short time and this PCR-based diagnostic system provides a simple and accurate method for detecting the presence of iridovirus infection.

• Fisheries and Aquatic Sciences
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• v.21 no.3
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• pp.5.1-5.8
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• 2018

Stimulator of interferon gene (STING) is induced by various inflammatory agents, such as lipopolysaccharide and microbial pathogens, including virus and bacteria. In this study, we obtained a full-length cDNA of a STING homolog from olive flounder using rapid amplification of cDNA ends PCR technique. The full-length cDNA of Paralichthys olivaceus STING (PoSTING) was 1442 bp in length and contained a 1209-bp open reading frame that translated into 402 amino acids. The theoretical molecular mass of the predicted protein sequence was 45.09 kDa. In the PoSTING protein, three transmembrane domains and the STING superfamily domain were identified as characteristic features. Quantitative real-time PCR revealed that PoSTING expressed in all the tissues analyzed, but showed the highest level in the spleen. Temporal expression analysis examined the significantly upregulated expression of PoSTING mRNA after viral hemorrhagic septicemia virus (VHSV) stimulation. In contrast, no significant changes in the PoSTING expression were detected in Edwardsiella tarda-challenged group compared to the un-injected control. The expression of P. olivaceus type I interferon (PoIFN-I) was also highly upregulated upon VHSV challenge. These results suggest that STING might be involved in the essential immune defense against viral infection together with the activation of IFN-I in olive flounder.

• Development and Reproduction
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• v.22 no.4
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• pp.341-350
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• 2018

Chemokines is a small protein that plays a major role in inflammatory reactions and viral infections as a chemotactic factor of cytokines involved in innate immunity. Most of the chemokines belong to the chemokine groups CC and CXC. To investigate the immune system of the olive flounder (Paralichthys olivaceus), an expression pattern specifically induced in the early developmental stages of analysis is examined using qRT-PCR. We also examined tissue-specific expression of both CC and CXC chemokine in healthy olive flounder samples. CC and CXC chemokine shows increased expression after immune-related organs are formed compared to expression during early development. CC chemokine was more highly expressed in the fin, but CXC chemokine showed higher expression in the gills, spleen, intestines, and stomach. Spatial and temporal expression analysis of CC and CXC chemokine were performed following viral hemorrhagic septicemia virus (VHSV) infection. CC chemokine showed high expression in the gills, which are respiratory organs, whereas CXC chemokine was more highly expressed in the kidneys, an immune-related organ. These results suggest that CC and CXC chemokine play an important role in the immune response of the olive flounder, and may be used as basic data for the immunological activity and gene analysis of it as well as other fish.

• Journal of fish pathology
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• v.19 no.3
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• pp.207-214
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• 2006

Diagnostic monitoring in fish farms with land-based tanks and net cases was conducted in eastern, western, southern and Jeju island of Korea for the summer period from 2000 to 2006. Total 3,518-fish samples of marine and freshwater fishes in 25 fish species were tested for pathogens. Fish species tested were olive flounder (Paralichthys olivaceus), fleshy prawn (Fenneropenaeus chinensis Osbeck), black rockfish (Sebastes schlegeli), rock bream (Oplegnathus fasciatus), red sea bream (Pagrus major), black seabream (Acanthopagrus schlegeli), sea bass (Lateolabrax japinicus), gray mullet (Mugil cephalus), rainbow trout (Onchorhynchus mykiss) and others. The infection rates by bacterial pathogens in the years of 2000, 2001, 2002, 2003, 2004, 2005 and 2006 were 22.4%, 34.5%, 14.1%, 15.3%, 17.7%, 13.5% and 5%, respectively. The infection rates by parasitic pathogens were 20%, 33.8%, 12.4%, 14.1%, 9.2%, 10.5% and 10.7%, respectively. The infection rates by viral pathogens were 22.4%, 13.5%, 10.3%, 5.4%, 9.7%, 10.2% and 15.8%, respectively. The infection rates by mixed pathogens were 10.3%, 0%, 44.9%, 50.9%, 31.9%, 38.4% and 39.6%, respectively. The rates of mixed infections were very low until 2001. The rates were higher than those of singer infections from 2002 to 2006. During the diagnostic monitoring from 2000 to 2006, the main bacterial pathogens were Vibrio (41.2%) and Streptococcus (28.8%). The infection rate by protozoa (85.7%) mainly including Scuticociliates and Trichodina was highest. The infection rate by viral necrosis virus (VNNV, 42.2%) was the highest of the viral pathogens.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.53 no.3
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• pp.432-442
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• 2020

Diagnostic monitoring in Korean rockfish cages was performed to survey the prevalence of pathogens in cultured Korean rockfish Sebastes schlegeli from May 2013 to July 2016. A total of 1,945 fish samples collected from the western (Cheonsu Bay and Heuksando), southern (Tongyeong and Namhae), and eastern coasts (Pohang) of Korea were tested for parasites, viruses, and bacteria. In this study, 1,264 and 334 fishes were infected with Microcotyle sebastis and Clavinema mariae, respectively. The prevalence rates of C. clavinema in fishes from Cheonsu Bay, Heuksando, and Tongyeong were 35.3%, 3.9% and 1.9%, respectively. No C. clavinema infection was detected in cultured rockfish from Namhae and Pohang. Furthermore, bacteria including Photobacterium damselae (8.9%), Photobacterium piscicola (2.3%), Photobacterium spp. (8.9%), Aeromonas salmonicida (1.8%), Aeromonas spp. (0.9%), Vibrio scophthalmi (1.5%), Vibrio spp. (3.3%), Streptococcus iniae (1.2%), and others (8.0%) were detected in 373 of 1,364 fishes. No virus was detected in any fish investigated in this study.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.49 no.2
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• pp.176-183
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• 2016

In this study, disease surveillance was performed to monitor the prevalence of viral haemorrhagic septicaemia virus (VHSV) and red seabream iridovirus (RSIV) in olive flounder, Paralichthys olivaceus in 2015. The fish samples were collected in March (60 farms), May (55 farms) and July (52 farms) from different farms in Jeju. Reverse transcription polymerase chain reaction (RT-PCR) (VHSV) or PCR (RSIV) results showed that VHSV detected in 2 farms, but RSIV has not been detected in any farms. The sequences of the nucleocapsid protein (N) and glycoprotein (G) gene of the 2 VHSV isolates were successfully sequenced. Phylogenetic analysis was included VHSV isolates reported here together with a representative VHSV isolates available in GenBank. Phylogenetic analysis indicated that most of Korea VHSV isolates were closely related to the Japan and China genotype IVa which is clearly distinct from the North American genotype IVb.

• Journal of Fisheries and Marine Sciences Education
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• v.27 no.3
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• pp.879-889
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• 2015

The outbreak of viral diseases caused by viral haemorrhagic septicaemia virus (VHSV) and red seabream iridovirus (RSIV) have been reported in cultured olive flounder, Paralichthys olivaceus. VHSV has been a serious viral disease that infects the olive flounders in South Korea. Clinical signs of VHSV infection are skin darkening, abdominal distension and haemorrhages. Outbreaks of fish iridoviral disease was first reported from red seabream, Pagrus major farms in Japan. Recently, iridovirus infection have occurred frequently from olive flounder farms in South Korea. In this study, disease surveillance was performed to monitor the prevalence of VHSV and RSIV in olive flounder in 2014. The samples were collected from 60 different olive flounder farms in Jeju from April, May, September, November and December in 2014. RT-PCR (VHSV) or PCR (RSIV) results showed that VHSV were detected in 5 farms, but RSIV has not been detected in any farms. The migration of olive flounder was restricted for the quarantine in 5 farms of VHS outbreak. The nucleocapsid protein (N) gene and glycoprotein (G) gene sequences of the 5 Korean VHSV isolates were successfully amplified and sequenced. Phylogenetic analysis was performed using the VHSV sequences reported here together comparison with the nucleotide sequences available from the GenBank database. Phylogenetic analysis indicated that most of Korea VHSV belong to the genotype IVa and closely related to the strains from Japan and China.

• Journal of fish pathology
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• v.34 no.1
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• pp.23-29
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• 2021

Interferon regulatory factors (IRFs) are a family of transcription factors essential to the control of antiviral immune response, cell growth, differentiation and apoptosis. IRF10 of zebrafish (Danio rerio) was negative regulation of the interferonΦ1 and 3 response in vitro. In this study, we analyze the induction of in vivo immune response activation from the IRF10 gene of zebrafish and the protective effect against VHSV. As the results, the group inoculated with IRF10 expression vectors, there was no expression of IFNΦ1, suggestion that IRF10 may function as a negative regulator of IRF3, which binds to the IFNΦ1 promoter. And other types of interferon genes (IFNΦ2-4) are thought to have been activated, inducing to the expression of pro-inflammatory cytokine and Mx genes. As the results of challenge test performed at 14 days after inoculation of the expression vectors, the maximum survival rate [50% (1㎍ DNA) and 42.5% (10㎍ DNA)] for IRF10 group were recorded. Meanwhile, the survival rates of pcDNA3.1 and PBS as the control groups were 10% and 15%, respectively. This study suggests that the possibility that activation of IRF10 molecule could be exploited as a VHS control method.

• Korean Journal of Environmental Biology
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• v.27 no.4
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• pp.391-396
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• 2009

The biological effects of carcinogenic polycyclic aromatic hydrocarbons (cPAHs) including benzo[a]pyrene (BaP), dibenzo[a,h]anthracene (DBA), benzo[a]anthracene (BaA), benzo[b] fluoranthene (BbF), benzo[k]fluoranthene (BkF), and indeno[1,2,3-c, d]pyrene (InP) on transcriptomic changes were determined in the liver of Oryzias latipes. Differentially expressed genes (DEGs) by binary exposure to cPAHs (BaP+BaA, BaP+BbF, BaP+BkF, BaP+DbA, BaP+InP) were screened by annealing control primers-based polymerase chain reaction followed by sequence analysis and BLAST searching. The results showed that four DEGs were commonly expressed by cPAHs and they were identified as ribosomal protein S4, coagulation factor II, elongation factor 1 beta, and a predicted protein similar to human immunodeficiency virus type I enhancer binding protein 3. This finding suggests that binary exposure to cPAHs interferes protein synthesis required for fundamental liver functions in fish.

• Development and Reproduction
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• v.21 no.4
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• pp.361-370
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• 2017

Koi herpesvirus (KHV), also known as Cyprinid herpes virus 3 (Cyprinid 3) is lethal disease in common carp and koi (Cyprinus carpio). Two different groups (KK and RK) were infected KHV by intraperitoneal injection. Fish for gene expression analysis were sampled at 0 h, 12 h, 24 h, 48 h and 72 h post infection (p.i). The results showed that two immune related gene, Interferons (INFs) ${\alpha}{\beta}$ and Interleukin (IL)-12 p35 induced a high response in RK. The IL-12 p35 cytokine and Toll-like receptor (TLR) 9 were significantly high expressed on 48 h post infection (p.i) in RK as compared to the KK. The histopatological examination reveals focal necrosis in liver and infiltrate of lymphocytes in spleen of KK as compared to the RK. In immunohistochemistry analysis, the KHV protein high expressed in the infected kidney cell and slenocyte of KK. Therefore, the expression of IL-12 p35, IFN ${\alpha}{\beta}$ and TLR 9 may provide a potentially genes related with KHV resistance in Koi and red common carp ${\times}$ koi.