• Journal of fish pathology
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• v.28 no.2
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• pp.113-116
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• 2015

A $0.45-{\mu}m$ membrane filter is generally used to remove bacterial contamination during virus extraction from fish samples. However, the number of fish viruses is drastically reduced after filtration with a $0.45{\mu}m$ filter. In this study, we investigated the effect of filters on virus infectivity titer and the change in virus titer and bacterial number following different centrifugation conditions to determine a suitable procedure for virus extraction from fish samples. $10^{4.05}$ and $10^{5.05}TCID_{50}/ml$ of infectious hematopoietic necrosis virus (IHNV) and $10^{4.05}$ and $10^{4.55}TCID_{50}/ml$ of Oncorhynchus masou virus (OMV) were not detectable after filtration with two types of $0.45-{\mu}m$ filters, except the IHNV titer was reduced by about 10 fold after filter use (company A). No significant difference was found in the virus titer following centrifugation at $880{\times}g$ (30 min) or $3,500{\times}g$ (30 min), whereas IHNV and OMV titers were reduced by about 10 and 10-1000 fold by centrifugation at $14,000{\times}g$ (30 min) and $14,000{\times}g$ (10 and 30 min), respectively. A total of 97.7-99.9% Escherichia coli were eliminated by centrifugation at $880 {\times}g$ (30 min) and $3,500{\times}g$ (30 min). These results show that fish viruses were affected by filtering, even though the effect differed by virus species and filter type. Therefore, centrifugation at $3,500{\times}g$ (30 min) and use of medium with antibiotics may be useful for virus extraction along with a reduction in bacteria.

• Development and Reproduction
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• v.27 no.2
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• pp.67-75
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• 2023

Polymeric immunoglobulin receptor (pIgR) mediates the transfer of polymeric immunoglobulin to protect organisms and is one of the most important mucosal effectors. In this study, the developmental stage- and tissue-specific expression of pIgR were observed before virus inoculation in olive flounder. pIgR was gradually expressed until the formation of immune tissue, exhibiting high expression in the late juvenile period; thereafter, pIgR expression gradually decreased and exhibited high expression in the spleen and skin. Moreover, pIgR expression after viral hemorrhagic septicemia virus infection was high in the kidney and spleen tissues at high density and low at low density. The results of this study can provide a basis for future studies on breeding density, virus expression, and immune system studies in fish.

• 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.25 no.2
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• pp.117-121
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• 2012

The pathogenicity of viral hemorrhagic septicemia virus (VHSV) from olive flounder Paralichthys olivaceus was investigated with masu salmon Oncorhynchus masou fry. The cumulative mortality of fish challenged with FYeosu05 isolate at $10^{6.5}$ $TCID_{50}$/fish was 60%. No mortality was observed in fish challenged with the isolates at $10^{5.5}$ $TCID_{50}$/fish and in mock-challenged fish. The affected fish showed darkening of the body, expanded abdomen, pale gills and enlarged spleen. VHSV from $10^{6.3}$ to $10^{7.8}$ $TCID_{50}$/g-tissue was re-isolated from the dead fish. These results suggest that the VHSV from olive flounder is pathogenic to masu salmon fry, although with low virulence.

• Journal of fish pathology
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• v.23 no.3
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• pp.335-342
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• 2010

In 2008, mass mortality was observed in paradise fish Macropodus opercularis which was imported from Indonesia. PCR of these fish found positive for megalocytivirus and Mycobacterium sp., while an unidentified virus was culture-isolated using CHSE-214 cells. In the present study, we investigated characterization of the unidentified virus and its pathogenicity to determine whether the virus was the causative agent of the mass mortality of paradise fish. The unidentified virus induced cytopathic effect (CPE) with syncytia in CHSE-214 and other fish cells, BF-2, GF, SSN-1, FSP and FFN. The virus was resistant against treatments with IUdR, chloroform, acidity at pH 3, basicity at pH 11 and high temperature at $56^{\circ}C$ for 3h. By electron microscopy, the viral particles were spherical having a double capsid structure with approximately 65 nm in external diameter. Viral genome was composed of at least 10-segmented RNA with sizes ranging from 0.7 kb to 3.6 kb. Based on these characters, this virus can be classified into family Reoviridae. This reovirus did not cause any mortality in an artificial experiment conducted by injecting the virus to paradise fish. This indicates that the reovirus is not only responsible for the mass mortality of paradise fish in 2008.

• Korean Journal of Ichthyology
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• v.27 no.3
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• pp.183-188
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• 2015

Recently, mariculture of rainbow trout (Oncorhynchus mykiss) has been initiated in the coast areas of Korea. In the present study, we investigated the effect of fish viruses on mariculture of rainbow trout. The pathogenicity of infectious hematopoietic necrosis virus (IHNV) and infectious pancreatic necrosis virus (IPNV) isolated from freshwater rainbow trout was tested against major cultured marine fish species, including olive flounder (Paralichtys olivaceus), rock fish (Sebastes schlegeli), rock bream (Oplegnathus fasciatus), red seabream (Pagrus major) and sevenband grouper (Epinephelus septemfasciatus). The pathogenicity of marine birnavirus (MABV), hirame rhabdovirus (HIRRV) and nervous necrosis virus (NNV) isolated from marine fish species was also tested against rainbow trout. No mortality was observed in marine fish species challenged with IHNV or IPNV. However, olive flounder and rock bream were infected by IHNV and IPNV. A mortality of 8.3% was observed in rainbow trout challenged with HIRRV. The fish was infected by both MABV and NNV. These results suggest that the mariculture of rainbow trout might be affected by fish viruses.

• Fisheries and Aquatic Sciences
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• v.1 no.2
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• pp.187-191
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• 1998

Chemotherapy can not be applied for the control of fish viral diseases because viruses depend on host machinery for their replication. Although new control strategies including vaccination are under development, avoidance of virus introduction by rapid and correct diagnosis is the best way of fish viral disease control. Although observation of virus particles with an electron microscope is an easy method for virus detection, it take a few days for the sample preparation. In order to shorten the sample preparation time, microwave radiation was applied in the procedure. With this method, 15 seconds was enough for fixation of virus infected fish samples or cultured cells inoculated with infectious hematopoietic necrosis virus, which takes 2-4 hours with routine methods. Also four minutes was enough for polymerization of embedding resin which takes 24-48 hours with routine methods. Samples prepared with microwave were good enough for direct electron microscopic observation and immunogold labeling assay.

• Fisheries and Aquatic Sciences
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• v.8 no.3
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• pp.185-187
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• 2005

We tested the in vivo ability of a DNA chip to detect virus-specific genes from virus-infected olive flounder Paralichthys olivaceus and rainbow trout Oncorhynchus mykiss. Target cDNA was obtained from total RNA of virus infected cell lines by reverse transcription (RT) and was labeled with fluorescent dye (Cy5-dUTP). The results show the successful detection of infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicaemia virus (VHSV) genes in the virus-infected fishes.

• Journal of Microbiology
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• v.34 no.3
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• pp.253-269
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• 1996

Infection of fish cells with IHNV resulted in gradual increase in cytosolic free Ca$\^$2+/ concentration ([Ca$\^$2+/)] in CHSE, gradual decrease in [Ca$\^$2+/] in FHM, and no significant change in RTG cells. The degree of [Ca$\^$2+/] increase or decrease was dependent on the amount of infectious virus, and these [Ca$\^$2+/] variations were maximal at 16 hours after virus infection (p. i.) in both cell lines. When the fish cells were infected with inactivated IHNV, evident variation in [Ca$\^$2+/] was not observed. Thus, infectivity of IHNV appears to correlate with changes in [Ca$\^$2+/] in virus-infected cells. These IHNV-induced [Ca$\^$2+/] changes were partially blocked by cycloheximide, but not affected by cordycepin. It seems to be that virus-induced Ca$\^$2+/ variations were more related with protein synthesis than RNA synthesis. Various Ca$\^$2+/ related drugs were used in search for the mechanisms of the [Ca$\^$2+/], changes following IHNV infection of CHSE cells. Decreasing extracellular Ca$\^$2+/ concentration or blocking Ca$\^$2+/ influx from extracellular media inhibited the IHNV-induced increase in [Ca$\^$2+/], in CHSE cells. Similar results were obtained with intracellular Ca$\^$2+/ sources are important in IHNV-induced [Ca$\^$2+/] increase in CHSE cells.

• Journal of fish pathology
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• v.23 no.3
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• pp.273-280
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• 2010

To determine whether rock bream Oplegnathus fasciatus can be protected from rock bream iridovirus (RBIV) infection by intramuscular injection of long double-stranded RNAs (dsRNAs), we compared protective effect of virus-specific dsRNAs corresponding to major capsid protein (MCP), ORF 084, ORF 086 genes, and virus non-specific green fluorescent protein (GFP) gene. Furthermore, to determine whether the non-specific type I interferon (IFN) response was associated with protective effect, we estimated the activation of type I IFN response in fish using expression level of IFN inducible Mx gene as a marker. As a result, mortality of fish injected with dsRNAs and challenged with RBIV was delayed for a few days when comparing with PBS injected control group. However, virus-specific dsRNA injected groups exhibited no significant differences in survival period when compared to the GFP dsRNA injected group. Semi-quantitative analysis indicated that the degree of antiviral response via type I IFN response is supposedly equal among dsRNA injected fish. These results suggest that type I IFN response rather than sequence-specific RNA interference might involve in the lengthened survival period of fish injected with virus-specific dsRNAs.