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        수산 생물의 생산과 관리에 관한 기초연구 항체 감작 Staphylococcus aureus 에 의한 Coagglutination Test 기법을 이용한 Edwardsiellosis 의 신속 진단

        하재이(Jai Yi HA),손상규(Sang Gyu SOHN),허민도(Min Do HUH),정현도(Hyun Do JEONG) 한국수산과학회 1996 한국수산과학회지 Vol.29 No.5

        To avoid the self-agglutination of Staphylococcus aureus sensitized with rabbit antibody in the absence of antigen, we determined the optimum concentration of rabbit antibody for sensitization. It was analyzed by using three different kinds of S. aureus strains at various concentraions of antibody. The optimized coagglutination test using the S. aureus sensitized with rabbit antibody was applied to the diagnosis of edwardsiellosis in field and in laboratory. The presence of E. tarda as low as 10 ㎍/㎖ was detected by this method. Moreover, it showed good coagglutination results against several different forms of antigens such as FKC, EDTA or heat extracted antigen of E. tarda. E. tarda strains, isolated from the flounders suffering from edwardsiellosis in fields, showed some cross-reactions to the E. tarda 219 analyzed by both agglutination and coagglutination test with rabbit anti-E. farda 219 antibody. The degree of cross-reactions analyzed was enough to apply the coagglutination test for the diagnosis of edwardsiellosis in field. Thus, even 1,000 fold diluted tissue homogenate of infected flounder naturally contained enough E. tarda as an antigen to show good coagglutination with S. aueus sensitized with rabbit anti-E. tarda 219 antibody. The successful application of this method to the homogenate and heat extract of tissues from naturally or artificially infected flounder or tilapia proved that coagglutination test was a simple and rapid reliable dignostic technique suitable for using in laboratory and field without any special equipments.

      • KCI등재

        수산생물의 생산과 관리에 관한 기초연구 : ELISPOT 기법을 이용한 넙치의 항체생성 세포분석

        하재이(Jai Yi Ha),박준효(Jun Hyo Park),김명석(Myoung Sug Kim),정준기(Joon Ki Chung),정현도(Hyun Do Jeong) 한국수산과학회 1999 한국수산과학회지 Vol.32 No.4

        We examined the immune response in flounder, Paralichthys olivaceus, with immunization of formalin killed Edwardsiella tarda as an antigen. The ELISPOT-assay (enzyme-linked immunospot assay) was optimized technically and applied to count the number of total and specific antibody secreting cells (TASC and SASC) in lymphocytes of different lymphatic organs. Incubation of lymphocytes on 96 well plate for more than 2.5hrs came out enough time in ELISPOT-assay for counting the antibody secreting cells in the anterior kidney and spleen. However, too much of plate-coated antigen or rabbit anti-flounder immunogtobulin for SASC or TASC counting, respectively, was appeared to decrease the sensitivity of the assay system. Specificity of the system was also confirmed by the absence of TASC in lymphocytes treated with cycloheximide to prevent protein synthesis. The peak numbers of SASC appeared at wk 3 post immunization after that there was a sharp decrease and reached to almost zero at wk 7. In the spleen and kidney, the timing and numbers of SASC on peak response were concurrent without preferential organ distribution. The specific antibody level in the sera increased rapidly between wk 2 and 3 after immunization, i.e. like the speafic cellular response found with ELISPOT-assay on that pedod. However, the remained high level of specific serum antibody from wk 5 after immunization until the end of experiment was clearly distinguishable from the kinetics af SASC response decreased sharply.

      • KCI등재

        향어에서의 oxolinic acid 흡수와 배설속도

        정준기(Joon Ki Chung),정현도(Hyun Do Jeong),하재이(Jai Yi Ha),허민도(Min Do Huh) 한국어병학회 1992 한국어병학회지 Vol.5 No.2

        The absorption and excretion times of oxolinic acid(OX) used in farms as new aquatic antibiotics commonly were evaluated with determination of the effects of water temperature and feeding to parameters by using the bioassay technique. On the same time, antibacterial activity and the complex formation of oxolinic acid with serum proteins of two different fishes were compared to those oxytetracycline(OTC). With more than 10 times lower MIC values than those of OTC in the strains among 13 analyzed fish pathogens. OX did not show the decresed antibacterial activity by the binding of serum proteins in carp and tilapia. It implies more powerful potential of OX as aquatic medicine OTC. The serum concentration of OX after different administrations the oral, i.m., i.v and dipping methods were compared. The higher beginning concentration in serum and faster excretion times were obserbed in i.m. and dipping methods respectively. In the oral and i.m. administration, peak serum concentration after 24-48 hrs and slow excretion times demonstrated in both methods. These pharmacokinetic characteristics similar at 30℃ and 20℃ water temperature conditions, however, beginning serum concentration of OX in fish dipped in 50㎎/ℓsol after starvation for 2 wks was appeared lower than those of fed fish. It suggests the importance of biological condition of the gill or skin for absorption of antibiotics after dipping administration.

      • KCI등재

        나일 틸라피아에 대한 질병예방제 및 면역보조제로서의 PS-K 효과 분석

        허성회,정현도,허민도,하재이,박경현 한국어병학회 1997 한국어병학회지 Vol.10 No.1

        단백질 결합 다당류인 PS-K가 Edwardsiella tarda의 감염에 대한 나일 틸라피아(Oreochromis niloticus)의 저항력 증강 효과에 대한 능력분석을 실시하였다. PS-K를 0.1 ㎎/g 어체중의 농도로 처리한 나일 틸라피아에 대하여 여러 농도외 E. tarda로 공격 실험을 하였다. 저농도(1.2×10^7 그리고 1.2×10^8 cfu/틸라피아)의 공격에 대하여 PS-K 처리구는 100% 생존율을 나타내었고 대조구는 각각 50%와 100%의 폐사율을 보여 주었다. 그러나 고농도의 E. tarda(1.2×10^9 그리고 1.2×10^(10) cfu/틸라피아) 공격에 대해서는 약간의 수명 연장효과만 있었을뿐 대조구와 같이 100%의 폐사율을 보여 주었다. E. tarda(1.2×10^7 cfu/틸라피아) 공격 1주일 후 PS-K를 복강 투여한 나일 틸라피아는 100%의 생존율을 나타내었으나 대조구는 50% 생존율을 나타내어 PS-K가 가지고 있는 치료제로서의 잠재력을 보여 주었다. 그리고 PS-K 투여 1주일 후 주사된 E. tarda FKC에 대하여 보여준 나일 틸라피아의 높은 항체 반응은 PS-K가 어류의 특이 액성 면역반응에 관여하여 어류가 세균 감염시 높은 저항성을 갖게 하여 주는 것으로 나타났다. A protein-bound polysaccharide preparation, PS-K, isolated from Coriolus versicolor was evaluated for its ability to enhance resistance against Edwardsiella tarda, causal agent of classical edwardsiellosis in tilapia, Oreochromis niloticus. Fish treated with a dose rate of 0.1 ㎎ PS-K g^(-1) body weight were challenged by intraperitoneal injection of E. tarda with different concentrations. Against low burdens of injections(1.2×10^7 and 12 ×10^8 cfu/fish), PS-K treated fish did not show any mortality compared to 50% and 100% mortality of control groups, respectively. However there was no increase of resistance against challenge with high concentrations of E. tarda (1.2×10^9 and 1.2×10^(10)cfu /fish) except few days delaying of death. Tilapia injected with PS-K one day after intraperitoneal inoculation of E tarda(1.2×10^7 cfu/fish) showed 100% survival rate compared to control group of 50% survival rate. The result indicates the potential of PS-K as a prophylactic agent in aquaculture. The increased antibody response in fish received PS-K one week prior to FKC administration suggested the influence of PS-K on the specific humoral immunity and increased resistance against bacterial infection.

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