• Title/Summary/Keyword: Fish vaccine

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Genetic Variations of Outer Membrane Protein Genes of Vibrio harveyi Isolated in Korea and Immunogenicity of OmpW in Olive Flounder, Paralichthys olivaceus (한국에서 분리된 Vibrio harveyi 외막단백질의 유전적 차이와 넙치(Paralichthys olivaceus)에 대한 OmpW의 면역원성 분석)

  • KIM, Myoung-Sug;JIN, Ji-Woong;JUNG, Sung-Hee;SEO, Jung-Soo;HONG, Suhee
    • Journal of Fisheries and Marine Sciences Education
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    • v.27 no.5
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    • pp.1508-1521
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    • 2015
  • Vibrio harveyi is a pathogenic marine bacterium causing systemic symptoms resulting in mass mortalities in fishes and shrimps in aquaculture. Outer membrane proteins(OMPs) are related to the pathogenicity and thus good targets for diagnosis and vaccination for Gram negative bacteria. Recently vaccination strategies using the OMPs have been suggested to control vibriosis in several fish species. In this study, we have isolated V. harveyi from diseased marine fishes from different regions of Korea and investigated genetic variations of four OMP genes including OmpK, OmpU, OmpV and OmpW. Consequently, OmpK and U genes could be divided into 3 subgroups of type I, II, III and type A, B, C, respectively, without any correlation with geographical regions and species while OmpV and W were highly homologous. OmpW gene of V. harveyi FP4138 was fully sequenced and predicted the deduced amino acid sequence to form ${\beta}-barrel$ with hydrophobic channel. Indeed, the immunogenicity of recombinant OmpW produced in Escherichia coli was assessed by vaccinating flounder. As a result, the high antibody response with antibody titer of $4.2{\pm}0.7$ and protection with relative percent survival of 60% against artificial infection of V. harveyi were demonstrated. This result indicates that OmpW is a virulence related factor and it can be a vaccine candidate to prevent a high mortality caused by V. harveyi infection in olive flounder, Paralichthys olivaceus.

Monitoring of Emaciation Disease in Cultured Olive Flounder Paralichthys olivaceus in Jeju (2010-2013), Korea (제주의 양식 넙치(Paralichthys olivaceus)를 대상으로 한 여윔증 모니터링(2010-2013))

  • Kim, Seung Min;Jun, Lyu Jin;Park, Myoung Ae;Jung, Sung Hee;Jeong, Hyun Do;Jeong, Joon Bum
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.48 no.5
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    • pp.719-724
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    • 2015
  • In this study aiming at the cultured olive flounders in Jeju island of Korea, has performed a total of 216 cases of pathogen tests for the olive flounders which seemingly had the symptom of emaciation in 24 farms on the suspicion of its occurrence from June 2010 to December 2013 and intended to get the basic information about the pathogen. According to the survey results of the emaciation infection status of 24 farms in Jeju over the period of this survey, it was confirmed that 18 (75%) of 24 farms in Jeju are positive in the emaciation infection. Among them, as for the rate of infection per year, it was observed that they are 38% in 2010, 48% in 2011, 50% in 2012 and 60% in 2013, and over the period of this survey the infection rate in accordance with the temperature of water has observed a variety of incidence rates from the summer to the winter season. In addition, according to the results of emaciation infection status for each size, the equivalent showed a detection rate, 54% in 11-20 cm, 43.9% in 21-30 cm and 25.4% in over 31 cm. This result accounts for an important portion among the diseases of farmed olive flounders in Jeju, is considered to be one of the diseases which cause troubles in the farms for olive flounders on land and it is thought that it can be utilized as basic data in order to estimate emaciation which may occur in the similar size of the cultured olive flounders in Jeju island.

Isolation and expression analysis of stimulator of interferon gene from olive flounder, Paralichthys olivaceus

  • Ma, Jeong-In;Kang, Sunhye;Jeong, Hyung-Bok;Lee, Jehee
    • Fisheries and Aquatic Sciences
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    • v.21 no.3
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    • pp.5.1-5.8
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    • 2018
  • Stimulator of interferon gene (STING) is induced by various inflammatory agents, such as lipopolysaccharide and microbial pathogens, including virus and bacteria. In this study, we obtained a full-length cDNA of a STING homolog from olive flounder using rapid amplification of cDNA ends PCR technique. The full-length cDNA of Paralichthys olivaceus STING (PoSTING) was 1442 bp in length and contained a 1209-bp open reading frame that translated into 402 amino acids. The theoretical molecular mass of the predicted protein sequence was 45.09 kDa. In the PoSTING protein, three transmembrane domains and the STING superfamily domain were identified as characteristic features. Quantitative real-time PCR revealed that PoSTING expressed in all the tissues analyzed, but showed the highest level in the spleen. Temporal expression analysis examined the significantly upregulated expression of PoSTING mRNA after viral hemorrhagic septicemia virus (VHSV) stimulation. In contrast, no significant changes in the PoSTING expression were detected in Edwardsiella tarda-challenged group compared to the un-injected control. The expression of P. olivaceus type I interferon (PoIFN-I) was also highly upregulated upon VHSV challenge. These results suggest that STING might be involved in the essential immune defense against viral infection together with the activation of IFN-I in olive flounder.

Biochemical, Hematological Effects and Complications of Pseudosynanceia Melanostigma Envenoming

  • Babaie, Mahdi;Zolfagharian, Hossein;Zolfaghari, Mohammad;Jamili, Shahla
    • Journal of Pharmacopuncture
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    • v.22 no.3
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    • pp.140-146
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    • 2019
  • Objectives: Venomous fishes have different pharmacological effects and are useful. Among the venomous fish, stonefishes; especially Pseudosynanceia melanostigma has various pharmacological effects on the nervous, muscular and cardiovascular system of humans. In this study, toxicological characteristics, some blood effects, pharmacological and enzymatic properties of Pseudosynanceia melanostigma venom was investigated. Methods: Crude venom purified by using gel filtration chromatography and the molecular weights of the venom and its fractions were estimated. The approximate LD values of this venom were determinedand the effects of LD50 dose on the blood of rabbits were studied. Hemolytic and Hemorrhagic activity of the venom sample was determined. In this case coagulation tests were performed. Results: The LD50 of the Pseudosynanceia melanostigma crude venom was also determined to be $194.54{\mu}g/mouse$. The effect of two doses of LD50 showed a non-significant differences decrease in RBCs and MCV. In other cases, the results showed significant differences in WBC, Plt, Hb, MCH, MCHC and HCT; also it's showed a significant decrease. WBC count showed a significant increase with two doses of LD50 groups. The prothrombin time and partial prothrombin time were increased after venom treatment. As well as bleeding and clotting time were increased. According to the results, a minimum dose for Haemorrhagic effect $40{\mu}g$ was obtained. Conclusion: Venom of Pseudosynanceia melanostigma has inhibitory effect on platelet aggregation that can be used to design and develop of anticoagulant drugs.

The specific antibody response of olive flounder Paralichthys olivaceus to route of antigen administration and change in environmental conditions (항원의 투여방법 및 사육환경 변화에 따른 넙치의 특이항체 반응)

  • Kim, Wi-Sik;Han, Jong-Seok;Jang, Min-Seok;Seo, Han-Gil;Jung, Sung-Ju;Park, Jung-Jun;Kim, Seok-Ryel;Oh, Myung-Joo
    • Journal of fish pathology
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    • v.24 no.2
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    • pp.153-160
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    • 2011
  • The specific antibody response of olive flounder, Paralichthys olivaceus to change in rearing-environmental conditions post immunization with antigen (bovine serum albumin, BSA) and different routes of antigen administration were investigated by enzyme-linked immunosorbent assay (ELISA). To test the effect of routes of antigen administration, flounder were injected intraperioneally or intramuscularlly with 1 mg of BSA. In addition, to test the effect of change in environmental condition post immunization, flounder were injected intraperioneally with 1 mg of antigen, and then were exposed to acute thermal change (the water temperature (WT) was decreased from $21^{\circ}C$ to $15^{\circ}C$ within 30 min and maintained at $15^{\circ}C$ for 3 h), handling (fish were caught and subsequently held out of water for 1 min) or heavy oil (76 g/200 L for 2 days). Consequently, there was no significant difference between intraperioneal (IP) and intramuscular (IM) injections except at 10 days post-immunization. With these results, it suggests that both 1M and IP injections may be used as route of vaccination. Furthermore, no significant difference was observed in the antibody response among the groups exposed to heavy oil, handling, sudden drop of WT and positive control except at 10 days post-immunization. From these results, it was confirmed that specific antibody response was not affected by the above mentioned rearing-environmental conditions, suggesting that vaccination can be employed at changing rearing-environmental conditions.

Roles of Glyceraldehyde-3-Phosphate Dehydrogenase in Edwardsiella tarda Pathogenesis (Edwardsiella tarda의 glyceraldehyde-3-phosphate dehydrogenase가 병원성에 미치는 영향)

  • Yu, Jong-Earn;Oh, Young-Eun;Lee, Tae-Ho;Kang, Ho-Young
    • Journal of Life Science
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    • v.20 no.12
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    • pp.1743-1749
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    • 2010
  • A research group demonstrated that the 37 kDA protein of Edwardsiella tarda, a causing causative agent of edwardsiellosis in fish, exhibited high antigenicity in Japanese flounder. The research group also showed that the N-terminus amino acid sequences of the 37 kDa protein were mapped to the N-terminus of GAPDH (glyceraldehyde-3-phosphate dehydrogenase). Using degenerated primer sets based on the known N-terminus sequence, the corresponding E. tarda DNA was amplified and cloned. The nucleotide sequences of the cloned gene revealed high homology with a bacterial gene for GAPDH, as we was expected. The amino acid sequence of E. tarda GAPDH (etGAPDH) revealed a <70% similarity with GAPDH proteins in other Enterobacteriaceae. With the application of artificial protein overexpression system in Escherichia coli, the recombinant etGAPDH (rGAPDH) was produced and purified. In this study, Using the purified rGAPDH, the etGAPDH specific polyclonal antibody has been was generated using the purified rGAPDHin this study. The immunoblotting analyses demonstrated that the location of the GAPDH protein is located with the association of is associated with the envelops of E. tarda. The rGAPDH was administrated into Japanese flounder via IP route for evaluation of the protective ability. Although the specific antibody titer against etGAPDH was increased about 3-fold after 4 weeks post-vaccination, the survival rates of vaccinated Japanese flounder and the control group with wild type E. tarda was were 12.5% and 0%, respectively. Our results indicated that rGAPDH is immunoreactive antigen but that it will not generate protective immunity in Japanese flounder.

A Case of Clonorchis Sinensis with Multiple Low Echogenic Nodules in a Child (소아에서 진단된 다발성 저음영 결절 형태를 보인 간흡충증 1례)

  • Hwang, Young Jun;Han, Man Yong;Ha, Doo Hoe
    • Pediatric Infection and Vaccine
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    • v.8 no.1
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    • pp.123-128
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    • 2001
  • Clonorchiasis, known as the parasitic fluke in the intrahepatic bile ducts, occurs throughout Korea, Japan, Taiwan, and Vietnam with high frequency. Most of the infections follow the consumptions of raw or undercooked freshwater fish, which explains its rare occurrence to childhood even in those areas mentioned earlier above. The authors have experienced it with an 8 year old boy. Abdominal ultrasonogram revealed multiple low echogenic nodules in the right lobe of liver and lymph nodes proliferation around the porta hepatis area. And abdominal C.T. scan also found multiple low attenuated nodules with delayed enhancement in liver parenchyma with hepatomegaly and multiple conglomerated lymph nodes in porta hepatis. Both abdominal ultrasonogram and C.T. scan didn't show any dilatation of peripheral intrahepatic bile ducts. In repeated stool examination, Clonorchis sinensis eggs were found in feces and the result of the skin test for Clonorchis sinensis was positive. The patient had a history of having eaten raw pond smelt back in Cheongju a month before the symptom onset. On the basis of the patient's history and the fact that Clonorchis eggs were found in feces 3 weeks after eosinophilia and symptoms were noted, the authors estimated that the findings of the patient's abdominal sonogram and C.T. scan were the radiologic findings of the acute phase of clonorchiasis, of which no report had ever been made before. So the authors report a case of clonorchiasis with atypical radiologic findings in an 8 year old boy.

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Intracellular Signaling Pathway for Host Defense Mechanisms against Piscine Nervous Necrosis Virus (NNV) (어류신경괴사증바이러스(nervous necrosis virus, NNV) 감염에 따른 숙주의 방어기전관련 세포신호전달)

  • Kim, Jong-Oh
    • Journal of Life Science
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    • v.30 no.4
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    • pp.402-409
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    • 2020
  • Nervous necrosis virus (NNV) contains a bi-segmented viral genome, RNA1 (3.4 kb, RdRp), and RNA2 (1.4 kb, capsid protein) in a small particle (25 nm). Despite its extremely compact size, NNV has caused serious damage by infecting approximately 120 fish species worldwide since it was first reported in the late 1980s. In order to minimize the damage caused by NNV infection and develop effective vaccines, it is necessary to understand the intra cellular signaling system according to NNV infection. NNV infection induces cell cycle arrest at the G1 phase via the p53-dependent pathway to use the cellular system for its replication. Otherwise, host cells recognize NNV infection through the RIG-1-like receptor (RLR) signaling pathway to control the virus and infected cells, and then ISGs required for antiviral action are activated via the IFN signaling pathway. Moreover, apoptosis of infected cells is triggered by the unfolded protein response (UPR) through ER stress and mitochondria-mediated cell death. Cell signaling studies on the NNV infection mechanisms are still at an early stage and many pathways have yet to be identified. Understanding the various disease-specific cellular signaling systems associated with NNV infection is essential for rapid and accurate diagnosis and vaccine development.

The Morphological characteristics of four Korean Abalone species in Nordotis (국내연안에 분포하는 둥근전복속 4종의 형태적 특징 고찰)

  • Won, Seung-Hwan;Kim, Soo-Kang;Kim, Sun-Chan;Yang, Byung-Kyu;Lim, Bong-Soo;Lee, Je-Hee;Lim, Han Kyu;Lee, Jung-Sick;Lee, Jun-Sang
    • The Korean Journal of Malacology
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    • v.30 no.1
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    • pp.87-93
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    • 2014
  • The Morphological characteristics of four species of Korean Abalone species, Nordotis discus hannai, N. discus discus, N. gigantea, N. madaka were compared. then Phenotypic traits were characterized. we collected from coast of samchonpo, sacjon-si, Gyeongsangnam-do and jeju-do of korea 69 individual of Nordotis discus hannai from 2013 December to 2014 January, 180 individual of N. discus, 72 individual of N. gigantea and 54 individual of N. madaka respectively then morphological traits of theses species characteristics were compared. The relationship between Shell length (SL), shell breth (SB), shell height (SH) and total weigh (TW) was expressed by the following equation: SB = 0.6346SL + 3.9082 ($R^2$ = 0.8956), SH = 0.2399SL + 3.2609 ($R^2$ = 0.8024), $TW=0.0009SL^{2.5622}$ ($R^2$ = 0.8088) in the Haliotis discus hannai, SB = 0.7249SL + 1.8035 ($R^2$ = 0.9634), SH = 0.3115SL - 11.223 ($R^2$ = 0.8593), $TW=0.0001SL^{2.9696}$ ($R^2$ = 0.8956) in the H. discus discus, SB = 0.7730SL - 1.1931 ($R^2$ = 0.933), SH = 0.2082SL + 3.2627 ($R^2$ = 0.6927), $TW=0.0002SL^{2.8330}$ ($R^2$ = 0.8431) in the Haliotis gigantea, SB = 0.7513SL - 1.0951 ($R^2$ = 0.913), SH = 0.2618SL - 6.1538 ($R^2$ = 0.6927), $TW=0.0001SL^{2.9614}$ ($R^2$ = 0.9353) in the Haliotis madaka.