넙치 혈청의 보존 조건이 ELISA 결과에 미치는 영향

김위식 ( Wi Sik Kim ),장민석 ( Min Seok Jang ),김종오 ( Jong Oh Kim ),김두운 ( Du Woon Kim ),정성주 ( Sung Ju Jung ),김석렬 ( Seok Ryel Kim ),박명애 ( Myoung Ae Park ),오명주 ( Myung Joo Oh ) 한국어병학회 2009 한국어병학회지 Vol.22 No.2

넙치(Paralichthys olivaceus)의 immunoglobulin M에 대한 단클론 항체 생산

김위식 ( Wi-sik Kim ),김기홍 ( Ki-hong Kim ),김춘섭 ( Choon-sup Kim ),오명주 ( Myung-joo Oh ) 한국수산과학회 2017 한국수산과학회지 Vol.50 No.2

Immunoglobulin M (IgM) was purified from olive flounder Paralichthys olivaceus sera using mannan-binding protein (MBP) and protein L affinity columns (designated as MBPIgM and ProLIgM, respectively). A monoclonal antibody (MAb) against olive flounder IgM was produced. The MBPIgM and ProLIgM had apparent molecular weights of 77, 73, and 28 kDa in SDS-PAGE. Nine hybridomas secreting MAbs against olive flounder IgM were established: five MAbs for MBPIgM (1, 2, 3, 4, and 5) and four for ProLIgM (6, 7, 8, and 9). Western blotting indicated that seven MAbs recognized heavy (H; MAbs 1, 2, 3, 4, 5, 6, and 7) chains and one recognized light (L; MAb 9) chains of IgM, while MAb 8 did not recognize IgM. The results of enzyme-linked immunosorbent assay (ELISA) with bovine serum albumin (BSA, antigen) and the nine MAbs revealed that the optical density (OD) values of sera differed significantly between BSA- and non-immunized fish, despite some sera from non-immunized fish with slight high OD values. These results suggest that the MAbs produced in this study reacted specifically with the IgM from olive flounder.

기후변화에 기인한 연안 수온상승에 따른 연안 어류의 바이러스성 질병 발생 예측

김위식 ( Wi Sik Kim ),김석렬 ( Seok Ryel Kim ),박명애 ( Myoung Ae Park ),이준수 ( Joon Soo Lee ),사티샤에분제 ( Satheesha Avunje ),김도형 ( Do Hyung Kim ),오명주 ( Myung Joo Oh ) 한국수산과학회 2013 한국수산과학회지 Vol.46 No.5

The impact of global warming on aquatic systems has been a priority research area in the past decade. However, the possibility that increased temperatures will cause shifts in viral disease outbreaks has not been well addressed. In the present study, with increasing water temperature (WT) in the coastal area of Korea, we estimated the possibility of changes in fish viral diseases. From the present time, WT may rise between 0.62 and 1.7 ℃ by 2050, and the effect on aquaculture could be more adverse than benefitial. Red seabream iridovirus disease (RSIVD) and viral nervous necrosis (VNN) cause high mortality above 22 and 24 ℃, respectively, and outbreaks could commence earlier and persist for prolonged periods. Nevertheless, the period of occurrence of viral hemorrhagic septicemia (VHS), which outbreaks at a lower WT (<18℃), could be shorter than the current infectious period. Thermal stress in fish causes reductions in growth and immunocompetence; thus, increases in summer WT can lead to the development of new viral diseases. WT has a strong influence on fish population dynamics; therefore, entry of new viruses and changes in the prevalence of infection can be expected if carrier fishes are introduced or migrate to Korean waters.

시판 전복박리제의 박리 효과

김위식 ( Wi Sik Kim ),강민호 ( Min Ho Kang ),김종오 ( Jong Oh Kim ),이시우 ( Si Woo Lee ),김정 ( Jung Kim ),황두진 ( Doo Jin Hwang ),오명주 ( Myung Joo Oh ) 한국어병학회 2013 한국어병학회지 Vol.26 No.2

Three commercial exfoliating reagents, product A (main components: citric acid and vitamin C), B (herb) and C (nicotinamide), were used to study their exfoliation effect on abalone, Haliotis discus hannai from the substrate. The exfoliating reagents A, B, C and oxytetracycline (OTC, control) of 6-31.25 g/L and 12-37.5 g/L exfoliated 81.2-84.8% and 90.3-95% of abalone, respectively. Post-treatment recovery time for the abalone was similar for all the reagents except product C. Recovery period for the abalone immersed in OTC for 5-20 second was slightly shorter than the reagents A, B and C; however, no mortality was observed in any group except with the reagents B (concentration: 20 g/L, immersion time: 5 sec) and C (12 g/L, 10 and 20 sec) that showed negligible mortality of 3.3%. Higher concentration and longer treatment with the reagents resulted in longer recovery time of the detached abalone. Although abalone exposed to the reagents needs slightly longer time to recover than that to OTC, the exfoliation effect is much similar. These results indicate that the commercial exfoliating reagents can replace OTC to detach abalone, though they need to be cautiously handled.

조피볼락(Sebastes schlegeli )에 대한 비정형 Aeromonas salmonicida 포르말린 사균 백신의 효과

김위식 ( Wi-sik Kim ),이현호 ( Hyeon-ho Lee ),오명주 ( Myung-joo Oh ),한현자 ( Hyun-ja Han ) 한국어병학회 2018 한국어병학회지 Vol.31 No.1

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.

해수 사육 무지개송어(Oncorhynchus mykiss)의 위팽창증후군

김위식 ( Wi-sik Kim ),공경희 ( Kyoung-hui Kong ),오명주 ( Myung-joo Oh ) 한국어류학회 2017 韓國魚類學會誌 Vol.29 No.1

본 연구에서는 2014년 제주도에 위치한 양식장에서 해수로 사육 중인 무지개송어에서 만성질병으로 인해 약 10%의 누적 폐사가 발생하였다. 병어는 복부가 팽만되어 있었고 위가 심하게 팽창되어 있었다. 질병 검사 결과, 기생충, 세균, 바이러스는 검출되지 않았으나 팽창된 위에서 다수의 Candida sp.가 분리 되었다. Candida sp.를 사용하여 감염 실험을 실시한 결과, 대부분 개체에서 폐사가 발생하지 않았고 위 팽창 증상이 관찰되지 않아 Candida sp.는 본 질병의 원인 병원체가 아닌 것으로 사료되었다. 본 연구에서는 국내에서 처음으로 해수 사육 무지개송어에서 위팽창증후군이 발생하는 것을 확인하였다. About 10% mortality occurred in cultured rainbow trout Oncorhynchus mykiss at a marine farm in Jeju in 2014. Diseased fish showed markedly abdominal distension and distended stomach. Although parasites, bacteria or viruses were not isolated from diseased fish, numerous Candida sp. were isolated from distended stomach. In experimental infection with Candida sp., mortality was not observed in most fish and clinical sign of distended stomach was not observed in the fish. These results suggest that Candida sp. may not be the etiologic agent of stomach distension syndrome. This is the first report of stomach distension syndrome in seawater farmed rainbow trout in Korea.

식용작물을 이용한 전복 박리

김위식 ( Wi-sik Kim ),김종오 ( Jong-oh Kim ),오명주 ( Myung-joo Oh ) 한국어병학회 2017 한국어병학회지 Vol.30 No.1

본 연구에서는 29종의 식용작물을 이용하여 전복박리 효과를 조사한 결과, 스피아민트, 레몬밤, 로즈플라워, 양파, 와사비분, 겨자분, 오미자 및 히비스커스 용액에서 박리효과가 관찰되었다. 특히, 오미자와 히비스커스 용액에서는 90% 이상의 박리율이 관찰되었다. 양식장에서 사육중인 전복을 대상으로 오미자 및 히비스커스 용액을 사용하여 전복 박리 및 회복 효과를 조사한 결과, 3%와 4%의 오미자 용액에 노출된 전복은 각각 93.6%(850/908)와 97.1% (810/834)의 박리율을 보였으며 1분 15초 이내에 회복되었다. 3%와 4%의 히비스커스 용액에서는 각각 96.1% (780/812)와 97.4%(700/719)의 박리율을 보였고, 박리된 전복은 1분39초 이내에 회복되었다. 이상의 결과, 오미자와 히비스커스 용액은 양식현장에서 전복을 박리하는데 실용적으로 사용 가능할 것으로 사료된다. Twenty nine edible plants were selected in order to study their exfoliation effect on abalone Haliotis discus hannai from its substrate. Exfoliation was observed in spearmint Mentha spicata, lemon balm Melissa officinalis, rose flower, onion Allium cepa, wasabi powder, mustard powder, omija Schizandra Chinensis, and hibiscus Hibiscus sabdariffa solutions. In particular, both the omija and hibiscus solutions exfoliated over 90% of the abalone. In field tests, 3% and 4% omija solutions exfoliated 93.6% (850/908) and 97.1% (810/834) of abalone, respectively, whereas 3% and 4% hibiscus solutions exfoliated 96.1% (780/812) and 97.4% (700/719), respectively. The detached abalone were recovered within 1 min 39 seconds. These results indicate that the omija and hibiscus solutions (3% and 4%) may be useful in the exfoliation of abalone.

넙치의 미동정 진균 기생증

김위식 ( Wi Sik Kim ),백근식 ( Keun Sik Baik ),김두운 ( Du Woon Kim ),서중경 ( Joong Kyeong Seo ),오명주 ( Myung Joo Oh ),강소영 ( So Young Kang ) 한국어류학회 2015 韓國魚類學會誌 Vol.27 No.4

본 연구에서는 2014년 넙치 치어를 사용하여 실험하는 과정에서 폐사가 발생하였다. 병어는 외관상 특이한 증상을 보이지 않았으나 아가미에서 많은 양의 균사가 관찰되었다. 기생충과 세균은 병어로부터 분리되지 않았다. 진균에 감염된 넙치와 건강한 넙치를 혼합 사육한 결과, 건강한 넙치는 7일이내에 100% 폐사되었고 폐사어는 아가미에 진균이 심하게 감염되어 있었다. 병리조직학적 검사 결과, 아가미의 새엽, 새판 및 새파 주위에 균사체와 다수의 균사가 관찰되었다. 이상의 연구 결과, 넙치의 폐사는 진균 감염과 연관성이 있을 것으로 사료되었다. In 2014, high mortality was observed in olive flounder (Paralichthys olivaceus) during experimental infection. Diseased fish did not show any outward clinical signs, but numerous fungal hyphae were detected in the gills. No parasites or bacteria were isolated from the diseased fish. A high mortality rate (100%) resulted when cohabitation with fungi-infected fish were applied to healthy fish. The affected fish exhibited sever fungi infection in the gills. Histopathological examination revealed numerous fungal hyphae and mycelium around gill filament, lamellar and racker. These results suggest that the fungi may be related with the mortality of olive flounder.

Infectious hematopoietic necrosis virus (IHNV)-검출 Reverse transcriptase Loop-mediated isothermal amplification (RT-LAMP) 법의 평가

김위식 ( Wi Sik Kim ),전찬혁 ( Chan Hyeok Jeon ),김정호 ( Jeong Ho Kim ),오명주 ( Myung Joo Oh ) 한국어병학회 2012 한국어병학회지 Vol.25 No.3

A reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assay was evaluated to monitor infectious hematopoietic necrosis virus (IHNV) from artificially infected rainbow trout Oncorhynchus mykiss. The cumulative mortalities of fish challenged with IHNV at 106.5 TCID50/fish, 105.5 TCID50/fish and 104.5 TCID50/fish were 40%, 0% and 0%, respectively. Dead fish and survivors at 16 and 28 d post-challenge in each group were employed for IHNV detection by RT-LAMP assay and virus isolation using BF-2 cells. IHNV from 104.3 to 106.8 TCID50/㎖ was isolated from all the dead fish and also detected in all of the examined dead fish by RT-LAMP assay. In survivors at 16 d, 60% (3/5 fish, 102.8-105.05 TCID50/㎖), 20% (1/5 fish, 101.05 TCID50/㎖) and 60% (3/5 fish, 101.05-104.8 TCID50/ ㎖) were found to be IHNV-positive by virus isolation in fish challenged with IHNV at 106.5 TCID50/fish, 105.5 TCID50/fish and 104.5 TCID50/fish, respectively, while 20% (1/5 fish), 0% (0/5 fish) and 20% (1/5 fish) were IHNV-positive by RT-LAMP assay. No IHNV was detected in the survivors at 28 d and control fish. These results indicate that the RT-LAMP assay is useful for detection of IHNV in diseased fish although it is not enough to monitor virus in IHNV-survivors.

넙치 (Paralichtys olivaceus) 유래 viral hemorrhagic septicemia virus (VHSV, genotype IVa)의 무지개송어 (Oncorhynchus mykiss) 감염 실험

김위식 ( Wi-sik Kim ),정하나 ( Ha-na Jeong ),공경희 ( Kyoung-hui Kong ),김아라 ( Ah-ra Kim ),전영호 ( Young-ho Jeon ),오명주 ( Myung-joo Oh ) 한국어류학회 2016 韓國魚類學會誌 Vol.28 No.3

본 연구에서는 넙치 유래 VHSV (genotype IVa)에 의한 무지개송어의 영향을 조사하기 위하여 다양한 조건에서 감염 실험을 실시하였다. 3개의 다른 로트의 담수 무지개송어 치어에 VHSV (10^6.3-7.3 TCID₅₀/fish)를 복강 주사한 결과, 15% 이하의 누적 폐사율이 관찰되었다. 무지개송어 (in vivo)를 사용하여 VHSV를 계대 배양한 후 감염 실험을 실시한 결과, VHSV를 5회 계대 배양하여도 독력의 변화는 관찰되지 않았다. 해수에 사육 중인 송어 (성어)에 넙치 유래 VHSV(10^5.3 TCID₅₀/fish와 10^6.3 TCID₅₀/fish)를 복강 주사한 결과, 폐사어는 관찰되지 않았고, 어체 내에서도 VHSV는 검출되지 않았다. 이상의 결과, 해수 송어 (성어)에서는 넙치 유래 VHSV의 병원성이 확인되지 않았으나 담수 무지개송어 치어에서는 낮은 독력을 나타내었다. Experimental infection of rainbow trout Oncorhynchus mykiss with viral hemorrhagic septicemia virus (VHSV, genotype IVa) from olive flounder Paralichtys olivaceus was examined. The cumulative mortalities of three different lot of rainbow trout fry challenged with VHSV (10^6.3-7.3 TCID₅₀/ fish) were less than 15%. No difference of virulence was observed in experimental infection using 5 in vivo passaged VHSV and original VHSV. No mortality was observed in seawater-reared rainbow trout (adult) challenged with VHSV (10^5.3-6.3 TCID₅₀/fish) and virus was not detected in the fish. We thus concluded that VHSV from olive flounder has low virulence to rainbow trout fry, but not pathogenic to seawater-rainbow trout (adult).