• Title/Summary/Keyword: red sea bream iridovirus

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Detection of RSIV (Red Sea Bream Iridovirus) in the Cultured Marine Fish by the Polymerase Chain Reaction (중합효소연쇄반응 (Polymerase Chain Reaction, PCR)법을 이용한 남해안 양식 해산어의 Red Sea Bream Iridovirus (RSIV) 보유상황 확인)

  • Oh, Myung-Joo;Jung, Sung-Ju;Kim, Young-Jin
    • Journal of fish pathology
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    • v.12 no.1
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    • pp.66-69
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    • 1999
  • Occurrences of red sea bream iridovirus disease (RSIVD) in cultured marine fishes were investigated. The infection was detected by the polymerase chain reaction (PCR) used to amplify the red sea bream iridovirus (RSIV). The RSIV infection was widely distributed in fish culture farm around the south coastal area of the Korean peninsula.

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Pathogenicity of Iridovirus against Marine Fish and Its Detection in Culturing Seawater (Iridovirus의 해산 양식어류에 대한 병원성과 사육수에서의 검출)

  • Jeong, Joon-Bum;Jeong, Hyun-Do
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.41 no.1
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    • pp.20-25
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    • 2008
  • The susceptibility of five different marine fish to iridovirus IVS-1 infection was analyzed and found a higher the cumulative mortality in the order of rock bream (Oplegnathus fasciatus), red sea bream (Pagrus major), sea perch (Lateolabrax sp.), rockfish (Sebastes schlegeli) and black porgy (Acanthopagrus schlegeli). However, the concentrations of virus in the infected spleens of these species did not differ significantly. To determine the release of iridovirus from infected fish into culturing seawater, rock bream were challenged with iridovirus IVS-1 and the concentration of virus in the water was analyzed using PCR. Over the 10 days of the analysis, the linear relationship between the number of dead fish and viral DNA concentration found in culturing seawater should be considered direct evidence of horizontal iridovirus transmission.

Genetic relatedness of Megalocytivirus from diseased fishes in Korea (국내 어류에서 분리된 Megalocytivirus의 유전형 분류 및 상관관계 분석)

  • Lee, Eun Sun;Cho, Miyoung;Min, Eun Young;Jung, Sung Hee;Kim, Kwang Il
    • Journal of fish pathology
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    • v.32 no.2
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    • pp.49-57
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    • 2019
  • In this study, we collected 39 megalocytiviruses isolated from diseased fish in Korea from 2012 to 2018. Major capsid protein (MCP) gene, a part of vascular endothelial growth factor (VEGF) gene and histidine triad motif-like protein (HIT) genes of Megalocytivirus were targeted for PCR amplification and analysis of those DNA nucleotide sequences. Korean strains revealed two genotypes (red sea bream iridovirus and turbot reddish body iridovirus types) based on the phylogeny of MCP gene. The red sea bream iridovirus type (RSIV-type) megalocytiviruses were divided into RSIV-subgroup 1 and 2. From the phylogenetic analysis of the VEGF genes, a genotypic variant of RSIV-type Megalocytivirus was identified. The HIT-like protein gene was detected in RSIVs, but not in TBRIV and ISKNV, suggesting that HIT-like protein gene may be specific in RSIV.

Characterization of rock bream (Oplegnathus fasciatus) fin cells and its susceptibility to different genotypes of megalocytiviruses

  • Jeong, Ye Jin;Kim, Young Chul;Min, Joon Gyu;Jeong, Min A;Kim, Kwang Il
    • Journal of fish pathology
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    • v.34 no.2
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    • pp.149-159
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    • 2021
  • Genus Megalocytivirus cause red sea bream iridoviral disease (RSIVD) and scale drop disease (SDD). Based on the phylogeny of the major capsid protein (MCP) and adenosine triphosphatase (ATPase) genes, megalocytiviruses except for SDD virus (SDDV) could be three different genotypes, red sea bream iridovirus (RSIV), infectious spleen and kidney necrosis (ISKNV), and turbot reddish body iridovirus (TRBIV). In this study, primary cells derived from the caudal fin of rock bream (Oplegnathus fasciatus) grew at 25℃ in Leibovitz's medium supplemented with 10% (v/v) fetal bovine serum and primocin (100 ㎍/mL). Rock bream fin (RBF) cells exhibited susceptibility to infections by different genotypes of megalocytiviruses (RSIV, ISKNV and TRBIV) with the appearance of cytopathic effects with an increase in the viral genome copy number. Furthermore, compared to grunt fin (GF) cells, even though 10 times lower number of RSIV genome copies were inoculated in RBF cells, viral genome copy number produced on RBF cells were 44 times higher than that of GF cells at 7 d post-inoculation. As the isolated RBF cells are sensitive to different genotypes of megalocytiviruses (RSIV, ISKNV and TRBIV), they can be used for future studies regarding in vitro viral infection and subsequent diagnosis.

Evaluation of a novel TaqMan probe-based real-time polymerase chain reaction (PCR) assay for detection and quantitation of red sea bream iridovirus

  • Kim, Guk Hyun;Kim, Min Jae;Choi, Hee Ju;Koo, Min Ji;Kim, Min Jeong;Min, Joon Gyu;Kim, Kwang Il
    • Fisheries and Aquatic Sciences
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    • v.24 no.11
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    • pp.351-359
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    • 2021
  • The red sea bream iridovirus (RSIV) belonging to genus Megalocytivirus is responsible for red sea bream iridoviral disease (RSIVD) in marine and freshwater fishes. Although several diagnostic assays for RSIV have been developed, diagnostic sensitivity (DSe) and specificity (DSp) of real-time polymerase chain reaction (PCR) assays are not yet evaluated. In this study, we developed a TaqMan probe-based real-time PCR method and evaluated its DSe and DSp. To detect RSIV, the probe and primers were designed based on consensus sequences of the major capsid protein (MCP) genes from megalocytiviruses including RSIV, infectious spleen and kidney necrosis virus (ISKNV), and turbot reddish body iridovirus (TRBIV). The probe and primers were shown to be specific for RSIV, ISKNV, and TRBIV-types megalocytiviruses. A 95% limit of detection (LOD95%) was determined to be 5.3 viral genome copies/µL of plasmid DNA containing the MCP gene from RSIV. The DSe and DSp of the developed real-time PCR assay for field samples (n = 112) were compared with those of conventional PCR assays and found to be 100% and 95.2%, respectively. The quantitative results for SYBR Green and TaqMan probe-based real-time PCR were not significantly different. The TaqMan probe-based real-time PCR assay for RSIV may be used as an appropriate diagnostic tool for qualitative and quantitative analysis.

Development of DNA Vaccine Against Red Sea Bream Iridovirus (RSIV)

  • PARK SO-JIN;SEO HYO-JIN;SON JEONG HWA;KIM HYOUNG-JUN;KIM YUN-IM;KIM KI-HONG;NAM YOON-KWON;KIM SUNG-KOO
    • Journal of Microbiology and Biotechnology
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    • v.15 no.4
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    • pp.873-879
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    • 2005
  • Red sea bream iridovirus (RSIV) obtained from infected rock bream was propagated by Bluegill fry-2 (BF-2) cell culture. The virus titer was determined as $10^{5.5}\;TCID_{50}/ml$ on confluent BF-2 cell monolayers. The integrin binding site of ORF 055L of infectious spleen and kidney necrosis virus (ISKNV) was selected for the construction of a primer to obtain the RSIV ORF 055L gene. The genes were amplified using RSIV gene lyzate by PCR. The homologies of the ORF 055L sequence of RSIV with ISKNV and rock bream iridovirus (RBIV) were approximately $96\%$ and $100\%$, respectively. DNA vaccine was constructed by cloning the ORF 055L of RSN into pcDNA 3.1 (+), containing a cytomegalovirus (CMV) promoter. For antibody production, pcDNA-055 DNA vaccine was injected to BALB/c mice. The production of antibodies against pcDNA-055 DNA vaccine was confirmed by the Western blot analysis. The antibodies produced by the pcDNA-055 DNA vaccine showed efficacy to neutralize the RSIV in the neutralization test in BF-2 cell culture.

Antiviral Efficacy of an Aquatic Disinfectant Tablet Composed to Calcium Hypochlorite Against Red Sea Bream Iridovirus

  • Cha, Chun-Nam;Lee, Yeo-Eun;Kang, In-Jin;Yoo, Chang-Yeul;Park, Eun-Kee;Kim, Suk;Lee, Hu-Jang
    • Journal of Food Hygiene and Safety
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    • v.27 no.3
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    • pp.285-289
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    • 2012
  • In this study, the veridical efficacy of an aquatic disinfectant tablet composed to calcium hypochlorite against red sea bream iridovirus (RBIV). A veridical efficacy was determined with the viability of RBIV contacted with the disinfectant in viral stock cultured in fat head minnow cell line. An aquatic disinfectant tablet and RBIV were reacted on the distilled water (DW), hard water (HW) or organic matter suspension (OM) condition. On DW and HW condition, RBIV was inactivated with 25,000 fold dilutions of an aquatic disinfectant tablet. With the investigation of the antiviral effect of the disinfectant on OM condition, RBIV was inactivated on 22,000 fold dilutions of an aquatic disinfectant tablet. As an aquatic disinfectant tablet possesses veridical efficacy against RBIV, the disinfectant solution can be used to limit the spread of cultured marine fish viral disease.

The First Report of a Megalocytivirus Infection in Farmed Starry Flounder, Platichthys stellatus, in Korea

  • Won, Kyoung-Mi;Cho, Mi Young;Park, Myoung Ae;Jee, Bo Young;Myeong, Jeong-In;Kim, Jin Woo
    • Fisheries and Aquatic Sciences
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    • v.16 no.2
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    • pp.93-99
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    • 2013
  • In 2009, a systemic megalocytivirus infection associated with high mortality was detected for the first time in cultured starry flounder Platichthys stellatus in Korea. Diseased starry flounder had pale bodies and gill coloring and enlarged spleens. Histopathological examinations revealed basophilic enlarged cells in various organs of diseased starry flounder. Polymerase chain reaction (PCR) was performed on tissue samples using three published primer sets developed for the red sea bream iridovirus. PCR products were detected for all primer sets, except 1-F/1-R, which are registered by the World Organization for Animal Health (OIE). The part of the gene corresponding to the full open reading frame encoding the viral major capsid protein (MCP) was amplified by PCR. PCR products of approximately 1,581 bp were cloned, and the nucleotide sequences were analyzed phylogenetically. The MCP gene of the starry flounder iridovirus, designated SFIV0909, was identical to that of the turbot reddish body iridovirus (AB166788).

Experimental transmission of red sea bream iridovirus (RSIV) between rock bream (Oplegnathus fasciatus) and rockfish (Sebastes schlegelii)

  • Min, Joon Gyu;Jeong, Ye Jin;Jeong, Min A;Kim, Jae-Ok;Hwang, Jee Youn;Kwon, Mun-Gyeong;Kim, Kwang Il
    • Journal of fish pathology
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    • v.34 no.1
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    • pp.1-7
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    • 2021
  • Red sea bream iridovirus (RSIV), belonging to the genus Megalocytivirus, is the predominant cause of mortality in marine fishes in Korea, including rock bream (Oplegnathus fasciatus). Rockfish (Sebastes schlegelii) are the host fish for RSIV, exhibiting no clinical signs or mortality. Cohabitation challenges, which mimicked natural transmission conditions, were performed to evaluate viral transmission between rock bream and rockfish, and to determine the pathogenicity and viral loads. In cohabitation challenge, artificially RSIV-infected rock bream were the viral donor, and healthy rockfish were the recipient. The results showed that although the donor rock bream had 95-100 % cumulative mortality (>108 viral genome copies/mg of spleen 7-14 days after viral infection), the recipient rockfish did not die, even when the viral genome copies in the spleen were >105 copies/mg. These results indicated asymptomatic infections. Notably, in a reverse-cohabitation challenge (artificially RSIV-infected rockfish as the viral donor and healthy rock bream as the recipient), RSIV horizontally infected from subclinical rockfish to rock bream (107 viral genome copies/mg of spleen 21 days after cohabitation) with 10-20% cumulative mortality. These results suggest that an asymptomatic, infected rockfish can naturally transmit the RSIV without being sacrificed.

Phylogenetic analysis and antigenic determinant prediction of red sea bream iridovirus isolated in Korea from 2019 to 2023 (2019년부터 2023년까지 국내에서 분리된 참돔이리도바이러스의 계통 분류 및 항원 결정기 예측)

  • Guk Hyun Kim;Joon Gyu Min;Hyun Do Jeong;Kwang Il Kim
    • Journal of fish pathology
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    • v.37 no.1
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    • pp.25-36
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    • 2024
  • In this study, we analyzed the phylogenetic classification, epitope prediction, and pathogenicity of red sea bream iridovirus (RSIV) isolated from rock bream between 2019 and 2023. Phylogenetics based on genes encoding MCP and ATPase indicated that all five RSIV isolates belonged to RSIV subtype II. The deduced amino acid sequence of the MCP for the amplicons (1362 bp) obtained from RSIV isolates had a length of 453 amino acids. Among these, the amino acid sequences of the RSIV-19, 21, 22, and 23 isolates showed 100% identity, while the RSIV-20 isolate showed 99.78% identity with one residue difference at position 306. As a result of antigenicity analysis based on amino acid sequence, the antigenicity score of the RSIV-20 isolate was 0.6386 and the other RSIV isolates were 0.6365. Additionally, the prediction of their antigenic determinants resulted in a total of 17 identical antigenic plots. When each RSIV was inoculated into rock bream, no significant differences were observed with 100% cumulative mortality in all groups. This study provides data on the potential for genetic variation of RSIV isolated in the same marine area over the past five years, and the antigenicity and pathogenicity results of each isolate are expected to be useful information for selecting future vaccine strains.