• Journal of fish pathology
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• v.31 no.1
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• pp.35-39
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• 2018

Atypical furnculosis caused by atypical Aeromonas salmonicida, is an emerging problem of farming of rockfish (Sebastes schlegeli) in Korea. In this study, protection against atypical furunculosis was compared in rockfish vaccinated with A. salmonicida formalin killed cell (FKC) and adjuvant containing FKC. The formalin inactivated A. salmonicida vaccine provided a low protection of 20% and 10% relative percent survival (RPS) at 44 and 58 days post vaccination. However, addition of adjuvant (squalene and aluminum hydroxide) into inactivated A. salmonicida vaccine clearly enhanced the level of protection showing 70% and 50% RPS at 44 and 58 days post vaccination.

• Journal of Aquaculture
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• v.21 no.4
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• pp.325-330
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• 2008

Two different Eelwardsiella tarda isolates, KFE and Edk-2, were obtained from Korea and Japan respectively. On the base of the previous results showing higher pathogenicity of E. tarda KFE compared to that of E. tarda Edk-2 isolate, we tried to determine the differences of antigenicities of these two isolates in loach which is one of the important species in freshwater aquaculture in Korea. Concentration of specific antibody in the serum appeared to be much higher in loach immunized with FKC of E. tarda Edk-2 than those found in loach immunized with FKC of E. tarda KFE. Cross-reaction analysis using agglutination test with normal and antigen-absorbed antisera implied the differences epitopes in the antigens of these E. tarda isolates. For the comparison of bactericidal activity of the produced antibody with different antigens, absorption analysis was performed and confirmed the presence of critical epitopes in the FKC of E. tarda KFE strain. The prophylactic agent, polysaccharide-bound protein (PS-K) injected 1 week before the artificial infection with E. tarda KFE isolates decreased the cumulative mortality in loach and would be on effective method to prevent the occurrence of bacterial infection including E. tarda.

• Journal of fish pathology
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• v.10 no.2
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• pp.75-86
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• 1997

Solid phase enzyme linked immunosorbent assay (ELISA) was developed to detect the whole cells of Edwardsiella tarda from infected tissues of flounder. Cross-reaction test was performed by ELISA against fish pathogens such as A. hydrophila ATCC7966. V. anguillarum HYUFP5001, Y, ruckeri 11-4, E. ictaluri and Streptococcus sp. NG8206. Rabbit anti-E, tarda Edk-2 sera highly cross-reacted with A. hydrophila ATCC7966 and V. anguillarum HUFP5001. However, the cross-reaction was removed by using the anti-serum pre-adsorbed with A, hydrophila ATCC7966 FKC. The intra-species cross-reaction among E. tarda isolates was very high. ELISA with the whole cell antigens present in tissue homogenate appeared with highly decreased sensitivity, presumably by the co-coating of lipid or proteins in tissues. Thus, it would be necessary to use the infected tissue homogenates diluted more than 100 times with PBS for diagnosis. Interestingly, compared with the using of FKC antigen, the direct detection of viable cells in tissue homogenate showed more sensitive results with detection limit of $1{\times}10^3$ cells/ml in buffer or diluted tissue homogenate. Consequently, the ELISA method developed in this study was specific, rapid and sensitive for diagnosing edwardsiellosis.

• Journal of Fisheries and Marine Sciences Education
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• v.26 no.6
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• pp.1366-1372
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• 2014

This study was conducted to select the media for the formalin killed vaccine (FKC) production of Vibrio harveyi and its application for olive flounder, Paralichthys olivaceus. For this, we have investigated the immune effects of Vibrio harveyi FKC vaccines grown in 3 different media i.e. Tryptic Soy Broth (TSB), TSB containing 2-2'-dipyridyl (TSB-D), Brain Heart Infusion Broth (BHIB) on the production of agglutinating antibody and protection against Vibrio harveyi in olive flounder. Additionally, a dual vaccine was prepared by combining Streptococcus parauberis vaccine to V. harveyi vaccine and its efficacy was also analyzed with the determination of optimal administration dosage. Consequently, olive flounder immunized with FKC grown in TSB-D showed the same protection with the vaccine grown in BHIB and the optimal dose of the vaccine was 10mg/kg of body weight. Indeed the dual vaccine showed higher agglutination titer and protection than control fish. The optimal dose for dual vaccine was 10mg/kg body weight for each of two vaccines.

• Journal of Microbiology and Biotechnology
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• v.19 no.9
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• pp.982-986
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• 2009

For the production of ghost bacteria vaccine to prevent the streptococcal disease in aquaculture fish species, a double cassettes vector was constructed and cloned in Escherichia coli DH5${\alpha}$. Ghost bacteria vaccine production from Escherichia coli DH5${\alpha}$/pHCE-InaN-GAPDH-Ghost 37 SDM (SIG) was maximized at a glucose concentration of 1 g/l, agitation of 300 rpm, and aeration of 1 vvm. The maximal efficiency of ghost bacteria formation was obtained at the mid-exponential phase ($OD_{600}=2.0$) with the concentration of 0.77 g/l for SIG. The molecular mass of GAPDH was detected at 67 kDa with the insoluble fraction, by SDS-PAGE and Western blot. The protective efficacy of ghost bacteria vaccine was evaluated by challenge test using olive flounder. The cumulative mortalities of the positive control, formalin-killed cell (FKC) vaccine, and SIG vaccine immunized groups were 91%, 74%, and 57%, respectively. These results suggest that SIG vaccine showed efficacy as a vaccine and had a higher potential to induce protective antibodies than did FKC vaccine.

• Journal of fish pathology
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• v.7 no.2
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• pp.127-149
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• 1994

Histopathological studies on the two lymphomyeloid organs of spleen and head kidney in tilapia, Oreochromis niloticus, were carried out to clarify the significance on the morphological characteristics of melano - macrophage centers (MMCs) which are varied in different physiological and pathological conditions of teleosts. To examine the histological changes by the artificial modification of the immunological states, tilapia were treated intraperitoneally with FKC and LPS of Edwardsiella tarda, and orally with dexamethasone, and then followed by the intraperitoneal injection of colloidal carbon for chasing the macrophages. There were marked differences in phagocytic avidity of macrophages, and accumulating patterns of carbon - ladening macrophages into the MMCs among the test groups. In the non - pretreated control group, carbon - ladening macrophages were densely accumulated at 12th and 20th day within the MMCs of head kidney and spleen, respectively. And, in the groups treated with bacterial antigens (FKC & LPS), the macrophages were more rapidly and densely aggregated within MMCs. But in the group with dexamethasone, only a few carbon particles were detected in both organs. Any compactly isolated form of particles was not found in this group. From the present results, it was strongly suggested that certain changes in immunological states of tilapia influence on the morphology of MMCs including the frequency of appearance, sizes, aggregating patterns or outlines. Therefore, morphology of MMCs would be very important in the interpretation for histopathological findings seen in the teleost's lymphomyeloid organs.

• Journal of fish pathology
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• v.19 no.1
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• pp.73-82
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• 2006

Streptococcal infections were considered as a serious problem because of significant economic losses in fish farm industry. We evaluated the efficacies of Streptococcus iniae vaccines in olive flounder, Paralichthys olivaceus. The vaccines were prepared from 10% neutral buffered formalin to give a final concentration of 0.3% or 3%, respectively. Fish were immunized by intraperitoneal injection of the experimental vaccines once or twice. Neither of the vaccines gave rise to any significant side effects. The antibody titers of booster immunized groups were significantly higher than those of prime immunized groups with both of the vaccines. According to formalin dosage, significantly increased antibody titers were produced by 3% formalin-killed cells (FKC) at 4weeks and 8weeks after prime and booster vaccination, respectively. Although the different levels of antibody production were showed by the vaccinated fish, the good protection obtained in challenge trials of the both vaccines. Fish immunized with 0.3% FKC once or twice had the relative percent survival (RPS) of 66.7% and 87.5%, respectively. Similarly, fish immunized with 3% FKC once or twice had the RPS of 70.0% and 77.0%, respectively. Further experiments are needed to study not only correlation between the antibody titers and RPS against S. iniae but also the side effects of high dose of formalin on antigenicity.

• Journal of fish pathology
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• v.19 no.3
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• pp.285-288
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• 2006

The respiratory burst activity of tilapia (Oreochromis mosambicus) phagocytes agaisnt Edwardsiella tarda ghosts produced by gene E mediated lysis was investigated. Stimulation with E. tarda ghosts (ETG) showed markedly higher respiratory burst activity than that with formalin killed E. tarda (FKC) in phagocytes. This result may suggest that ETG can induce effective cellular immune responses in fish.

• Journal of fish pathology
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• v.19 no.3
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• pp.243-251
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• 2006

The internal defense system of mollusks consists of circulating haemocytes. In order to understand the morphological characterization of haemocytes, light and electron microscopy were carried out in oyster, Crassostera gigas. Four types of haemocytes were recognized: type Ⅰ small hyalinocytes, type Ⅱ large hyalinocytes, type Ⅲ large granulocytes and type Ⅳ small granulocytes. Additionally, the activities of alkaline phosphatase (ALP), acid phosphatase (ACP), peroxidase (POD), α-naphthyl acetate esterase, β-glucuronidase, PAS, sudan black B and oil red O in haemocytes were analysed by immunocytochemical methods. The results indicate that enzymatic activities were abundant and more active in granulocytes than in hyalinocytes. After incubation with haemoctyes and Vibrio FKC, phagocytic index and percentage of phagocytic cell were and shown to be increased from 15 to 120 min. In addition, the enzymatic activities were higher than those of controls: ALP, ACP, α-naphthyl acetate esterase and β-glcuronidase, indicating that these enzymes can be related with phagocytosis in oyster.

• Journal of fish pathology
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• v.21 no.3
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• pp.201-208
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• 2008

We compared the pathogenicity and antigenicity of two different Edwardsiella tarda (E. tarda) strains KFE and Edk-2 isolated in Korea and Japan respectively. In the pathogenicity analysis with challenge test against loach, E. tarda KFE isolate showed stronger pathogenicity compared to that of E. tarda Edk-2 isolate. The differences were also confirmed by the comparison of OMP (outer membrane protein) in SDSPAGE which showed three major bands, 41kDa, 37kDa and 30kDa, for E. tarda KFE isolates and two major bands, 41kDa and 30kDa, for E. tarda Edk-2 isolates. On the base of these results, we tried to determine the differences of antigenicities of these two isolates in loach which is one of the important species in freshwater aquaculture in Korea. Numbers of specific antibody secreting cells (SASC), appeared to be higher in loach immunized with FKC of E. tarda Edk-2 than loach immunized with FKC of E. tarda KFE. ELISPOT-assay for the comparison of antigenicity showed relatively high percentage of cross-reaction and implied the presence of some common epitopes in the antigens of these two E. tarda isolates.