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신속한 항생제 감수성 테스트 법의 개발을 위한 Water-Soluble Tetrazolium Salt의 적용
황성돈 ( Seong Don Hwang ),조동희 ( Dong Hee Jo ),김광일,조미영 ( Mi Young Cho ),지보영 ( Bo Young Jee ),박명애 ( Myoung Ae Park ),박찬일 ( Chan Il Park ) 한국어병학회 2015 한국어병학회지 Vol.28 No.2
In this study, we conducted to the development of a rapid antimicrobial susceptibility test method using WST-1 which is known to water-soluble tetrazolium salt, in order to rapidly response against bacterial diseases in fish. Eight of antibiotics which are permissioned for marine organism from government were used to rapid antimicrobial susceptibility testing using the WST-1. As a result, a similar tendency was verified compare to conventional antibiotic susceptibility test results. Generally, the antibiotic susceptibility test method required about 3 days (72 hours) for determine the effective antibiotics, however, we have confirmed that the our method using WST-1 was required at least 36 hours in this study. Consequentially, our method will contribute to development of rapid antimicrobial susceptibility testing for bacterial diseases in fish.
2016년 소양호에서 채집한 자연산 잉어과 어류와 갑각류의 법정전염병 및 기생충성 질병 모니터링
문성희 ( Seong Hee Mun ),허준욱 ( Jun Wook Hur ),차승주 ( Seung Joo Cha ),황성돈 ( Seong Don Hwang ),손맹현 ( Maeng-hyun Son ),권준영 ( Joon Yeong Kwon ),권세련 ( Se Ryun Kwon ) 한국수산과학회(구 한국수산학회) 2018 한국수산과학회지 Vol.51 No.1
The presence of pathogens in wild fish is a potential threat to the fish being raised nearby fish farm. Surveillance of these pathogens in the wild is, thus, highly important to keep fish in the farm safe from serious communicable diseases. Fish and crustacean were sampled 4 times at Soyang Lake in 2016. Pathogens for five reportable communicable diseases including KHVD (koi herpesvirus disease), SVC (spring viraemia of carp), EUS (epizootic ulcerative syndrome) and WSD (white spot disease), and parasites were investigated. In Soyang Lake, pale chub Zacco platypus was the most abundant. Forty seven pale chub, 8 Korea piscivorous chub Opsarichthys uncirostris, 3 oily shiner Sarcocheilichthys variegatus wakiyae and 2 crucian carp Cyprinus carassius were investigated for detection of target diseases of fish. Seventeen Caridina denticulata denticulata, 12 Oriental river prawn Macrobrachium nipponense and 1 Chinese mitten crab Eriocheir sinensis were used for detection of WSD. As the result, reportable communicable diseases were not detected in all the freshwater animals. Dactylogyrus was detected in Korea piscivorous chub. Copepoda was also detected in pale chub and Korea piscivorous chub. Metacercaria was detected in the pale chub sampled in September and October. Those were not metacercaria of liver fluke Clonorchis sinensis.
바이러스성 출혈성 패혈증 바이러스(Viral Hemorrhagic Septicemia Virus) 감염 넙치(Paralichthys olivaceus)의 조직별 바이러스 정량분석
장진현,황성돈,정지민,권문경,황지연,Jang, Jin Hyeon,Hwang, Seong Don,Jung, Ji Min,Kwon, Mun-Gyoung,Hwang, Jee Youn 한국환경생물학회 2021 환경생물 : 환경생물학회지 Vol.39 No.3
국내외에서 80여 종이 넘는 담수 및 해산어류를 감염시켜 대량폐사를 발생시키는 바이러스성 출혈성 패혈증 바이러스(VHSV) 진단검사를 위해 넙치의 여러 조직의 바이러스 발현량에 대한 정량적 데이터를 시간순서에 따라 분석하였다. 무작위 선별된 넙치에 3.0E+07 TCID<sub>50</sub> per 0.1mL per fish의 VHSV를 복강 주사하여 시간순서(0시간, 6시간, 12시간, 1일, 2일, 3일, 5일, 7일)에 따라 조직(아가미, 간, 신장, 비장, 근육)을 채취하였다. Real-time PCR 법을 통해 상대 정량한 결과 5일차 아가미, 간, 신장, 비장에서 바이러스의 발현량이 가장 높게 나타났다. 이번 연구를 통해 감염 초기단계에서 비장이 VHSV 확정진단을 위한 적정조직임을 입증하였으며, 국내 법정전염병 진단에 중요한 정보를 제공할 것이다. A diagnostic test for viral hemorrhagic septicemia virus (VHSV), which infects more than 80 species of freshwater and marine fish at home and abroad, causing mass mortality, was conducted to provide quantitative data on the amount of virus expression in various tissues of flounder in chronological order. The tissues were collected in chronological order after the intraperitoneal injection of 3.0E+07 tissue culture infective dose<sub>50</sub> (TCID<sub>50</sub>) per 0.1mL per fish of VHSV to randomly selected flounder. As a result of relative quantification through real-time PCR, the highest levels of virus expression were found in the spleen, kidney, gill, and liver on day 5. This study proved that the spleen was an appropriate site for the final diagnosis of VHSV in the early stages of infection and will provide important information for the diagnosis of legal infectious diseases in Korea.
심재동 ( Jae-dong Shim ),황성돈 ( Seong-don Hwang ),장수영 ( Soo-young Jang ),김태완 ( Tae-wan Kim ),정지민 ( Ji-min Jeong ) 한국어병학회 2019 한국어병학회지 Vol.32 No.1
2015년부터 2017년까지 우리나라 넙치 양식장에서 발생하는 폐사피해를 표본조사를 통해 모니터링 하였다. 표본어가는 전국의 수산질병관리원과 지속적으로 거래하고 있으며 현장방문을 통해 설문이 가능한 양식어가 중 층화구조분석을 통해 선정 하였다. 수산질병관리사 조사요원의 직접적인 현장방문을 통한 설문 조사를 수행하여 객관적으로 양식현황 및 폐사 현황을 파악하였다. 조사기간 중 24.78%/2015년, 30.19%/2016년 그리고 21.59%/2017년의 누적 폐사율을 나타내었고, 감염성 질병에 의한 폐사는 스쿠티카증(2015-2017년 평균누적 폐사율: 56.7%)과 바이러스성출혈성 패혈증감염(추정)(2015-2017년 평균 누적 폐사율: 8.9%)에 의한 피해가 높은 것으로 나타났으며, 제주에서는 여윔증(2016-2017년 평균 누적 폐사율: 10.3%)에 의한 피해가 높게 나타났다. A monitoring was performed to survey the mortalities that had occurred in the aquaculture farms of olive flounder (Paralichthys olivaceus) in South Korea from 2015 to 2017. The indirect inquiry for entire farms and the sample survey for selected farms were carried out. The aquatic organism disease inspectors, who have a national license for the diagnosis and prevention of aquatic organism diseases and a have close relationship with the farms, investigated the rates and causes of mortalities according to the standard manual. The mortality rate by sample survey of farms in 2015, 2016, and 2017 were 24.78% (Chunnam: 17.86%, Jeju: 28.69), 30.19% (Chunnam: 24.45%, Jeju: 32.65), and 21.59% (Chunnam: 10.57%, Jeju: 26.00%), respectively. The major cause of mortality was scuticociliatosis, and the mortality caused by viral hemorrhagic septicemia and emaciation disease (Jeju) were also high. Our results can contribute to effective establishment prevention of epidemics system and acquired status as a disease-cleansing country.
Enzyme-linked immunosorbent assay를 이용한 바이러스성 출혈성 패혈증 바이러스 감염 넙치(Paralichthys olivaceus)의 특이 항체반응 검사
황지연,장진현,김동준,권문경,서정수,황성돈,손맹현,Hwang, Jee Youn,Jang, Jin Hyeon,Kim, Dong Jun,Kwon, Mun Gyeong,Seo, Jung Soo,Hwang, Seong Don,Son, Maeng-Hyun 한국수산과학회 2017 한국수산과학회지 Vol.50 No.5
The viral hemorrhagic septicemia virus (VHSV) has an extensive host range, and infects farmed and wild fish inhabiting both freshwater and marine ecosystems. Enzyme-linked immunosorbent assay (ELISA) is highly useful in diagnosing viral hemorrhagic septicemia. However, ELISA shows high, non-specific background reaction with fish antibodies. In this study, we optimized the antigen and antibody concentrations used for detecting specific antibodies in VHSV-infected olive flounder to reduce non-specific binding, and improve the sensitivity of ELISA. The results suggested that OD (optical Density) values were valid when ELISA was performed with $0.1{\mu}g/well$ of virus, involving blocking with blocking buffer (Roth, Roti-Block), 1:300-1:600 dilution with flounder antisera, and 1:1000 dilution with anti-flounder IgM and HRP-conjugated goat anti-mouse IgG for detecting the VHSV antibody in flounder sera. Furthermore, 11 different VHSV strains isolated in Korea from 2012 to 2016 were used to infect the fish. The results showed no correlation between viral pathogenicity and antibody production. This research is a basic study on the application of antibody detection in the diagnosis of viral hemorrhagic septicemia in the olive flounder.