• Journal of fish pathology
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• v.22 no.3
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• pp.219-228
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• 2009

The genomic and subgenomic RNAs of fish nodavirus encode the four proteins, protein A, capsid protein, non-structural protein B1 and B2. In this study, we describe the immune response of olive flounder Paralichthys olivaceus immunized with live fish nodavirus or recombinant capsid protein, non-structural protein B1 and B2 expressed in E. coli. Nodavirus-infected flounder produced antibodies to capsid protein, B1 and B2 and nodavirus-neutralizing activities were detected in the serum of the nodavirus-infected flounder. The flounder were immunized against the three recombinant proteins of fish nodavirus and the sera from these immunized fishes were assayed for nodavirus-specific antibody by ELISA and a neutralization test. In the immunized flounder, all three recombinant proteins induced the production of similar levels of antibody, but only the antibody to capsid protein significantly neutralized nodavirus. These results indicate that all three nodaviral proteins are immunogenic in flounder, but only the capsid protein can induce neutralizing antibody against nodavirus.

• Journal of fish pathology
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• v.12 no.2
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• pp.107-113
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• 1999

The pathogenicity of the fish nodavirus causing viral nervous necrosis (VNN) of sevenband grouper, Epinephelus septemfasciatus was examined in sevenband grouper and other marine fish by intramuscular injection. Sevenband groupers of 27~104 g in body weight were highly susceptible to the fish nodavirus, but yellowtail (537 g in body weight), red seabream (207 g), rock bream (43 g), flounder (41 g), tiger puffer (27 g) and rockfish (94 g) of the sizes used to this experiment were not. The pathogenicity of the viral agent to the sevenband grouper was high without regard to fish sizes at rearing water temperature over $20^{\circ}C$, but not at $15^{\circ}C$. Therefore, susceptibility of sevenband grouper to the viral agent causing VNN was thought to be water temperature-dependent rather than fish size (age-dependent). Compared to the infectivity of the viral agent to sevenband grouper with artificial infection methods, fish were successfully affected by intramuscular, intraperitoneal, oral, dipping and cohabitation administrations although there were slight differences in mortalities among infection methods. And survival sevenband grouper after infection with the fish nodavirus was resistant to the reinfection for a long time.

• Journal of fish pathology
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• v.11 no.2
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• pp.97-104
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• 1998

Since 1989, mass mortality has repeatly occurred in cage-cultured sevenband grouper, Epinephelus septemfasciatus along the southern coast of Korea in the summer season and usually reached over 80% within a few months. Diseased fish showed the clinical signs of anorexia, dark coloration, loss of eqilibrium, spinal swimming behaviour, vertebral deformity and inflation of swim bladder. Histopathologically, necrosis and/or vacuolation of the nerve cells in the brain and retina were observed. We previously reported that the causative agent was filtrable. The causative agent was not culturable in various fish cells; RTG-2, CHSE-214, BF-2, EPC and FHM. However, electron microscopic observation revealed unenveloped icosahedral viral particles with about 30 nm in diameter in the cytoplasm of nerve cells of the brain. The characteristics of the virus was tested by an artificial infection with the filtrate of the homogenate of diseased fish. The pathogenicity of the virus was retained after treatment with ether or heat ($50^{\circ}C$, 30 min) but partly lost by pH 3 or 11 treatment. These results suggest that the causative agent are similar to the fish nodavirus. In order to compare the causative agent with a fish nodavirus, Striped Jack Nervous Necrosis Virus (SJNNV), a polymerase chain reaction (PCR) was performed with primers specific to SJNNV. As a result, about 430 by PCR products were detected from the brain and the eye of both naturally and artificially infected sevenband grouper. All these results represent that the mass mortality in the cultured sevenband grouper is caused by the infection of a nodavirus similar to SJNNV and this is the first report of a fish nodavirus from the cultured sevenband grouper in Korea.

• Journal of fish pathology
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• v.33 no.1
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• pp.71-75
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• 2020

A survey was conducted to investigate viral infections in 184 whiteleg shrimp (Litopenaeus vannamei) collected from nine farms and one wholesale fish vendor during 2018 and 2019. Gill and abdominal muscle of shrimp were tested for the presence of five viruses, viz. infectious hypodermal and haematopoietic necrosis virus, taura syndrome virus, infectious myonecrosis virus, yellow head virus genotype 1, and covert mortality nodavirus by reverse transcription-polymerase chain reaction (RT-PCR) and PCR. These viruses were not detected in any of 184 samples, screened under the study.

• Journal of fish pathology
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• v.34 no.2
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• pp.141-147
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• 2021

In this study, of the imported shrimps between 2017 and 2020, we investigated white spot syndrome virus (WSSV), covert mortality nodavirus (CMNV) and decapod iridescent virus 1 (DIV-1). Of the imported shrimps (a total of 29 groups), WSSV was detected as 31% (9/29) by nested PCR assay. And CMNV and DIV-1 were not identified in this study. To investigate the genetic relatedness of WSSV identified from imported shrimp, VR 14/15 region showed WSSV genomic variable loci was compared with reference isolates. Among the nine WSSV-positive samples, VR 14/15 region was amplified in only a sample (20-CH-1 isolate, imported from China in 2020). And the 20-CH-1 isolate showed 99.8% identity with WSSV-IN-05-01 which was reported in India in 2005, suggesting that those of WSSV have been spread from India to China. Furthermore, although the pathogenicity of WSSV identified from frozen shrimp was not evaluated, the international trade of diseased frozen shrimps could be led to the potential risk of virus transmission.

• Journal of fish pathology
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• v.37 no.1
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• pp.1-8
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• 2024

The advantages of replicon vectors of RNA viruses include a high ability to stimulate innate immunity and exponential amplification of target mRNA leading to high expression of foreign antigens. The present study aimed to construct a DNA-layered nervous necrosis virus (NNV) replicon vector system in which the capsid protein gene was replaced with a foreign antigen gene and to compare the efficiency of foreign antigen expression between the conventional DNA vaccine vector and the present replicon vector. We presented the first report of a nodavirus DNA replicon-based foreign antigen expression system. Instead of a two-vector system, we devised a one-vector system containing both an NNV RNA-dependent RNA polymerase cassette and a foreign antigen-expressing cassette. This single-vector approach circumvents the issue of low foreign protein expression associated with the low co-transfection efficiency of a two-vector system. Cells transfected with a vector harboring hammerhead ribozyme-fused RNA1 and RNA2 (with the capsid gene ORF replaced with VHSV glycoprotein ORF) exhibited significantly higher transcription of the VHSV glycoprotein gene compared to cells transfected with either a vector without hammerhead ribozyme or a conventional DNA vaccine vector expressing the VHSV glycoprotein. Furthermore, the transcription level of the VHSV glycoprotein in cells transfected with a vector harboring hammerhead ribozyme-fused RNA1 and RNA2 showed a significant increase over time. These results suggest that NNV genome-based DNA replicon vectors have the potential to induce stronger and longer expression of target antigens compared to conventional DNA vaccine vectors.