• Fisheries and Aquatic Sciences
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• v.5 no.1
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• pp.32-35
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• 2002

In this study, we investigated the mechanism of cell death in rhabdovirus-infected cells, chinook salmon embryonic cell line (CHSE-2l4) infected with hirame rhabdovirus (HIRRV). Studies using light microscopy, fluorescence microscopy, TUNEL method, electron microscopy, and agarose gel electrophoresis revealed changes in the cell morphology and DNA fragmentation indicative of apoptosis in early infection. It was observed that HIRRV induced apoptosis as well as necrosis in infected cells.

• Journal of Microbiology and Biotechnology
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• v.15 no.2
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• pp.354-361
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• 2005

Vibrio fluvialis, an enteropathogenic bacterium, produces a phospholipase which is thought to be an important factor in the pathogenesis of disease. In this study, the phospholipase gene (vfp) was identified from V fluvialis (KCTC 2473) and its sequence was determined. The entire open reading frame was composed of 1,689 nuc1eotides and 563 amino acids. The phospholipase gene (vfp) was overexpressed in Escherichia coli as a his-tag fused protein. This recombinant protein (rVFP58) was solubilized with 6 M urea and purified by Ni-NTA affinity chromatography. The action mode of rVFP58 was determined by TLC and GC-MS, and it showed phospholipase B activity, which had both phospholipase A and lysophospholipase activities. The rVFP58 showed a maximum activity at pH around 9- 10 and temperature of about 40OC, and it was stable under alkaline condition over pH 9. The cytotoxicity of rVFP58 was evaluated, using a fish cell line, CHSE-2l4, and was found to cause significant cell death after 14 h of exposure to 250 $\mu$g of the protein.

• Journal of fish pathology
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• v.11 no.2
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• pp.129-136
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• 1998

In March 1997, a new rhabdovirus was isolated from moribund cultured Japanese flounder Paralichthys olivaceus in sea water tank and cage culture systems in Kyung-Nam and Chun-Nam province, Korea. At temperature $15^{\circ}C$ the virus replicated and induced cytopathic effects (CPE), which progressed to eventual cytolysis, in susceptible cell lines, including RTG-2 and EPC. The CHES-214 cell line was refractory. Virus particles were bullet-shaped and measured $70nm{\times}100$ to 150 nm in size. The isolate was sensitive to pH 3, to diethyl ether, and to heat ($50^{\circ}C$ 5 min, $60^{\circ}C$ 1 min). Viral replication was not inhibited by $10^{-4}$ M 5-iododeoxyuridine. Virus infectivity was reduced by anti-HRV (8401-H) rabbit serum, but can not reduced by antisera against infectious hematopoietic necrosis virus (IHNV), chum salmon reovirus (CSV), retrovirus of salmonid (RVS) and infectious pancreatic necrosis virus (IPNV). HRV virus antigen was detected by fluorescent antibody test (FAT) in the cytoplasm of infected EPC cell. Purified isolates virions were composed of: polymerase (L), glycoprotein (G), nucleoprotein (N) and 2 matrix proteins (M1 and M2). Based upon their relative mobilities, the estimated molecular weights of the proteins were: L, 160 kDa; G, 55 kDa; N, 45 kDa; M1, 26 kDa; and M2, 22 kDa.

• Journal of fish pathology
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• v.18 no.2
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• pp.167-178
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• 2005

Streptococcus iniae is one of the major bacterial pathogens of cultured olive flounder, Paralichthys olivaceus in Korea. This study was carried out to investigate the interaction between morphological characteristics and immune response such as phagocytosis and serum killing activity. They were differentiated phenotypically into two groups, the viscous colonies and the non-viscous colonies, but the strains in both groups were similar in physiological and biochemical characteristics. Electron microscopic examination of the viscous form revealed thick, electron-dense exopolysaccharide materials surrounding polycationic ferritin-stained cells, while the exopolysaccharide material was absent around the non-viscous form of S. iniae. The viscous type strains were disclosed more virulent than those of non-viscous type for olive flounder. The viscous strains were resistant to normal serum killing activity, while the non-viscous strains were susceptible to bactericidal activity of normal serum from olive flounder. The viscous strains were more resistant to opsonophagocytosis and decreased the chemiluminescence response of head kidney macrophages of olive flounder. All of the tested non-viscous S. iniae strains were efficiently destroyed by the macrophage after phagocytosis. Both the viscous and the non-viscous strains invaded and replicated in cultured fish cell line (CHSE-2l4). This cellular invasion may contribute to the pathogenesis of invasive S. iniae infection.