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국내 여윔 넙치에서 검출된 점액포자충 Parvicapsula sp.의 정량적 분석
김승민,정준범,Kim, Seung Min,Jeong, Joon Bum 한국어병학회 2018 한국어병학회지 Vol.31 No.2
여윔증상이 나타난 양식장(farm-B 및 farm-C)의 넙치 및 여윔증상이 나타나지 않은 양식장(farm-A) 넙치의 각 내부 장기(신장, 장, 비장, 뇌 및 간)를 대상으로 점액포자충 Parvicapsula sp.의 양적 분석을 real-time PCR 방법을 사용하여 각각 실시하였다. 여윔증상을 보였던 farm-C의 넙치 신장에서 가장 높은 DNA copy number ($1.7{\times}10^7copies/mg$ tissue)를 보였고, farm-B의 넙치에서는 모든 내부 장기에서 낮은 수치가 나타났으며, farm-A의 넙치에서는 모든 내부 장기에서 음성 결과를 나타내었다. 동일한 시료를 사용한 PCR 및 병리조직학적 분석에서도 real-time PCR에서의 결과와 같은 양상을 보였다. Quantitative analysis of a myxosporean parasite, Parvicapsula sp. in internal organs (kidney, intestine, spleen, brain and liver) from non-emaciated (farm-A) or emaciated (farm-B and farm-C) olive flounder Paralichthys olivaceus were performed by real-time PCR. The highest DNA copy number ($1.7{\times}10^7copies/mg$ tissue) was detected in kidney of the emaciated olive flounder from farm-C, while the DNA copy number was below detection limit in all the organs of the olive flounder from farm-B. There was not positive result in all of organs from olive flounder in farm-A. PCR and histopathological analysis were also performed using the same specimen and showed same results as those by real-time PCR.
Pearl Gourami (Trichogaster leeri)로부터 분리한 Iridovirus의 유전적 특성과 병원성 분석
김호열,정준범,전려진,윤소혜,조혜진,정현도,Kim, Ho Yeoul,Jeong, Joon Bum,Jun, Lyu Jin,Yoon, So Hye,Cho, Hye Jin,Jeong, Hyun Do 한국해양바이오학회 2006 한국해양바이오학회지 Vol.1 No.3
본 연구에서는 비장 내의 비대해진 세포 존재와 조직 괴사라는 병리조직학적 관찰에 의하여 우리 나라의 pearl gourami (Trichogaster leeri)에서 iridovirus에 의한 자연 감염이 나타남을 확인하였다. 이러한 iridovirus 감염을 더욱 정확하게 진단하기 위해서 iridovirus 감염 진단에 주로 사용되는 MCP와 ATPase gene 부위에서 2 primer sets를 제작하여 PCR을 실시한 결과, PCR 생성물은 기대한 size와 부합하게 나타났고, MCP gene 부위의 염기서열은 reference strain인 ISKNV와 매우 높은 유사성 (99.6%)을 보였다. 공격 실험을 통하여 pearl gourami에서 분리된 iridovirus의 병원성을 확인하였고, 무분별한 관상어 관리에 의하여 관상어로부터 양식어류에로의 질병 전이가 나타남으로서 일어날 수 있는 국내 양식 산업에 대한 위험성을 제시하였다. In the survey of the samples infected by iridovirus, we found the natural outbreak of iridovirus disease in the ornamental fish, pearl gourami (Trichogaster leeri) in Korea. It was characterized by the appearance of enlarged cells and necrosis in the observation of the imprinted spleen cells under iridovirus. To determine the infection of iridovirus more accurately, PCR using 2 different primer sets for MCP and ATPase gene used for the diagnosis of iridovirus infection was done and found the produced fragments matched with the size of expectation. Partially determined nucleotide sequences of the MCP gene of the iridovirus isolated in this study showed very high homology (99.6%) with that of ISKNV as a reference strain. In challenge experiment with the iridovirus isolated from the moribund of pearl gourami, the pathogenicity of the isolated iridovirus was confirmed and suggested the potential of the risk associated the transfer of iridovirus from the ornamental fishes to the marine culturing fishes.
Oxytetracycline과 neomycin 복합제의 약욕에 따른 양식 넙치(Paralichthys olivaceus)의 근육내 잔류량 변화
김승민,전려진,정준범,Kim, Seung Min,Jun, Lyu Jin,Jeong, Joon Bum 한국어병학회 2015 한국어병학회지 Vol.28 No.1
본 연구에서는 넙치 (평균체장 및 무게: $20{\pm}2.0cm$, $70{\pm}5.0g$)에 옥시테트라사이클린(oxytetracycline, OTC)-네오마이신(neomycin, N) 복합제를 25-10 ppm 및 50-20 ppm으로 용해하여 2일 동안, 3시간씩 약욕을 실시하였고, 약물투여 중지 후 1일, 5일, 14일, 40일째에 시험군별로 각각 5마리씩 넙치의 근육을 채취하여 항생제 잔류농도를 조사하였다. OTC와 N 표준용액을 농도별로 희석하여 표준곡선을 작성한 결과 각각 $R^2=0.9999$, $R^2=0.9952$로 양호한 직선성을 나타내었고, OTC와 N 회수율은 각각 90~93%, 88~95%가 관찰되었다. 잔류농도에 대한 측정결과, OTC와 N 복합제를 25-10 ppm 투여한 시험군에서 1일째 OTC는 $0.97{\pm}0.084{\mu}g/ml$이 검출되었고, N은 $0.118{\pm}0.079{\mu}g/ml$이 검출되었으나 5일째 이후부터 검출이 되지 않았다. OTC와 N 복합제를 50-20 ppm 투여한 시험군에서 1일째 OTC는 $1.324{\pm}0.062{\mu}g/ml$이 검출되었고 N은 $0.788{\pm}0.05{\mu}g/ml$이 검출되었으나 5일째 이후부터 검출이 되지 않았다. In the present study, we performed a dipping of olive flounder (average length and weight: $20{\pm}2.0cm$, $70{\pm}5.0g$) for a period of three hours a day, over two days, in a melted complex of oxytetracycline (OTC) and neomycin (N), by dissolving 25-10 ppm or 50-20 ppm in water. Subsequently, the remaining antibiotic density in muscle tissue collected from olive flounder was investigated, 1, 5, 14 and 40 days after discontinuation of the medication. 5 fish were used from each group. The standard graph drawn from the results of diluting two standard solutions of OTC and N based on various density levels, showed a relatively straight line with an $R^2$ of 0.9999 and 0.9952, respectively. The recovery rate of OTC was shown to be 90-93% and N, 88-95%. Upon measurement of the remaining antibiotic density in the test group that had been exposed to 25-10 ppm of the complex of OTC and N, $0.97{\pm}0.084{\mu}g/ml$ of OTC and $0.118{\pm}0.079{\mu}g/ml$ N were detected on 1 day of the test. No antibiotic density was detected after day 5 of the test. Regarding the test group that were exposed to 50-20 ppm of the complex of OTC and N, $1.324{\pm}0.062{\mu}g/ml$ of OTC and $0.788{\pm}0.05{\mu}g/ml$ N were detected on day 1 of the test, and no antibiotic density was detected after day 5 of the test.
여윔증 넙치, Paralichthys olivaceus의 증상에 대한 병태생리학적 고찰
김이경 ( Yi Kyung Kim ),정준범 ( Joon Bum Jeong ),이무근 ( Mu Kun Lee ),박수일 ( Soo Il Park ),박명애 ( Myeong Ae Park ),최미경 ( Mi Kyung Choe ),여인규 ( In Kyu Yeo ) 한국어병학회 2011 한국어병학회지 Vol.24 No.1
This study was aimed to investigate the pathophysiological changes of olive flounder, Paralichthys olivaceus suffering from emaciation. A plasma osmolality was higher in the emaciated and control flounders than that of normal teleost, suggesting osmoregulatory failure in both of them. Also, the control in the same stock with emaciated flounder seem to be classified into a primary degree of emaciation. According to microscopic observations, the inflammatory responses were observed in the submucosal layer of anterior intestine, although the some of mucosal intestinal epithelium still remained. It was suggested that the pathological changes of the anterior part give rise to malabsorption of nutrients through the mucosa. In the posterior intestine and rectum, the mucosal epithelium were almostly sloughed off and severe inflammatory responses were observed in the submucosa. Immunoreaction for NKCC was not detected in the mucosal epithelial cells in intestine because of sloughing of epithelium. These changes would lead to functional disorder in the intestine, such as malabsorption of nutrients and osmoregulatory failure. Also important is to investigate the recovery phase.
양식장 배출수 퇴적층에서 분리된 리파아제 생산 박테리아의 동정 및 배양학적 특성
김만철,장태원,장익수,여인규,정준범,허문수,Kim, Man-Chul,Jang, Tae-Won,Harikrishnan, Ramasamy,Jang, Ik-Soo,Yeo, In-Kyu,Jeong, Joon-Bum,Heo, Moon-Soo 한국미생물학회 2009 미생물학회지 Vol.45 No.1
제주 연안 양식장 배출수 퇴적층으로부터 미생물 배양 배지를 사용하여 총 200여 균주를 분리하였으며, 분리된 균주를 이용하여 olive oil이 함유된 평판배지에서 투명환을 형성하는 균주를 분리하였고, 이를 LI-68, LI-80로 각각 명명하였다. 분리 균주 LI-68, LI-80의 BIOLOG를 이용한 생화학적 분석 특성 및 16S rDNA의 염기서열 분석 결과 Janibacter anophelis와 99%의 유전적 상동성을 보여 최종적으로 Janibacter sp. LI-68, Janibacter sp. LI-80으로 동정되었다. 분리 균주 Janibacter sp. LI-68과 Janibacter sp. LI-80의 증식을 위한 최적 배양온도를 확인하고 증식온도에 따른 지방분해효소 활성의 변화를 조사하였으며, 이를 위하여 배양 온도를 $20^{\circ}C$에서 $40^{\circ}C$까지 $10^{\circ}C$ 간격으로 배양하여 균체의 증식과 분비되는 지질분해 효소의 활성을 조사하였다. 그 결과 분리균주 Janibacter sp. LI-68는 $30^{\circ}C$ 배양실험구에서 가장 높은 균 생육도를 나타냈으며, 효소활성은 균 생육도와는 다른 $40^{\circ}C$에서 가장 높은 효소활성을 보였다. 반면 Janibacter sp. LI-80 균주는 $30^{\circ}C$에서 가장 높은 균 생육도를 보였으며, 효소활성은 균 생육도와 반비례하여 $40^{\circ}C$에서 가장 높은 효소활성을 보였다. From the course of screening of useful enzyme producing microorganism from marine sedimentary layer, we isolated 2 lipase producing strains and their lipase producing activities were tested. 16S rDNA sequence analysis showed that they were Gram-positive bacteria grouped on Janibacter sp. An excellent lipase producing strain, Janibacter sp. LI-68 and J. sp. LI-80 identified by 16S rDNA analysis and biochemical methods (BIOLOG), was further studied its lipase producing characteristics. The optimum initial pH, temperature and the optimum cultral time for the enzyme production on MA medium were 8, $30{\sim}40^{\circ}C$ and 96 h, respectively.
양식 넙치에서 분리한 어병세균의 lincomycin에 대한 내성 유전자의 분포
김예지,전려진,이영준,고예진,한소리,김성현,정준범,Kim, Ye Ji,Jun, Lyu Jin,Lee, Young Juhn,Ko, Ye Jin,Han, So Ri,Kim, Sung Hyun,Jeong, Joon Bum 한국어병학회 2022 한국어병학회지 Vol.35 No.1
Lincomycin as one of the lincosamides antibiotics have been mainly used in human and livestock fields, but have not been used in aquaculture. In this study, the distribution of minimum inhibitory concentration (MIC) values against lincomycin and the detection of the macrolide-lincosamide-streptogramin (MLS) resistance gene were confirmed in bacterial pathogens isolated from cultured olive flounder (Paralichthys olivaceus). Of the 107 strains isolated from Jeju, 36 strains of Gram-positive bacteria and 71 strains of Gram-negative bacteria were identified. Most of Streptococcus spp. was found to have a MIC value of less than or equal to 0.5 ㎍/mL, and Edwardsiella piscicida was found to have a MIC value higher than 1,024 ㎍/mL. V. harveyi and V. alginolyticus mostly showed MIC values of 256 ㎍/mL, but V. scophthalmi displayed values of 8~64 ㎍/mL. In the detection of MLS resistance gene, erm(B) was detected in 9 strains of Streptococcus spp., and erm(A) was confirmed in one strain.