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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.
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
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.
Young-Chul KIM(김영철),Jee-Youn HWANG(황지연),Hae-Ryeon JEON(전해련),Da-Won LEE(이다원),Jung-Soo SEO(서정수),Kwang-Il KIM(김광일),Mun-Gyeong KWON(권문경),Bo-Young JEE(지보영),Seong-Don HWANG(황성돈) 한국수산해양교육학회 2018 水産海洋敎育硏究 Vol.30 No.6
We identified structural and non-structural gene regions encoding major capsid protein (MCP) and DNA polymerase (DPOL) of megalocytiviruses collected from infected cultured fishes in RBIVD outbreak farms in 2013-2017 in Korea. With the two PCRs using 1-F/1-R and 4-F/4-R primer sets of the Manual of Diagnosis Tests for Aquatic Animals of the World Organization for Animal Health (OIE), amplicons were generated from the spleen and kidney tissue from approxmately ~30 fishes, including rock bream (Oplegnathus fasciatus), red sea bream (Pagrus major), and rock fish (Sebastes schlegeli), from 15 outbreak regions in the aquatic farms of the South Sea and Jeju Island. In phylogenetic analysis, complete MCP and partial DPOL genes belonged to RSIV type-subgroup2. Interestingly, these genes formed a cluster indicating closer relatedness to GSIV-K1, RIE12-1, and RBIV-C1, which were previously isolated from Japan and China, than with RBIV-KOR-TY1 isolated from Korea. However, the nucleotide sequence identities of the MCP and DPOL genes of these viruses were high, at >99.8% and >99.7%, respectively, compared with RBIV-KOR-TY1. Comparisons of nucleotide and amino acid sequences showed minimal differences between the obtained strains in the MCP gene, however, one or two nucleotide sequences substitutions of the DPOL gene were detected in nine strains, including a silent mutation detected in five strains. These findings suggest a slow rate of evolution of megalocytiviruses in this region, but the potential for mutations and new pathogenic strains warrants continuous surveillance.
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.
수온변화에 따른 해수내 VHSV(Viral Hemorrhagic Septicemia Virus)의 생존능 분석
정지민(Ji Min JEONG),지보영(Bo-Young JEE),권문경(Mun-Gyeong KWON),서정수(Jung Soo SEO),황성돈(Seong Don HWANG),이지훈(Ji-Hoon LEE),황지연(Jee Youn HWANG) 한국수산해양교육학회 2019 水産海洋敎育硏究 Vol.31 No.5
Viral hemorrhagic septicemia (VHS) is a serious viral disease that infects the olive flounder in South Korea. The Korean aquaculture industry experienced an economic loss caused by the high infectivity and mortality. Wastewater is a factor in transferring infections to wild and cultured fish in waters near infected aquaculture farms. In this study, we analyzed the stability of the VHS virus in sterilized seawater depending on the water temperature. The VHS virus maintained infectivity for 8 days and was detected using quantitative polymerase chain reaction after 10 days of inoculation to sterilized seawater at low temperatures. On the other hand, at high temperatures, the virus maintained infectivity for 4 days and was detected for 7 days. These results may contribute to basic data for epidemiological studies.
서정수(Jung Soo SEO),이지훈(Ji-Hoon LEE),지보영(Bo-Young JEE),권문경(Mun-Gyeong KWON),황지연(Jee Youn HWANG),황성돈(Seong Don HWANG),정지민(Ji Min JEONG),심재동(Jae-Dong SHIM) 한국수산해양교육학회 2019 水産海洋敎育硏究 Vol.31 No.6
This research has studied current situation of aquatic medicine use of nine major aquaculture species in our nation from year 2016 to 2017. We have proceeded the research after selecting each species of domestic licensed aquaculture farms which used stratification structure samples(Flounder, 60 of sampling aquaculture farms/527 of total aquaculture farms; Rockfish, 63/819; Red sea bream, 60/463; Starry flounder, 37/79; Israeli mirror carp, 42/108; Eel, 57/360; Rainbow trout, 47/150; Whiteleg shrimp 51/209; Abalone 65/1,295). Classification of aquatic medicine research targets are divided into four big groups depending on their functional characteristics such as antibiotics, parasiticide, disinfectants and others(digestive medicine etc). We have visited each sampling aquaculture farms to check the amount of aquatic medicine that they have used. And the result came out in the order of Flounder> Rockfish> Red sea bream, Eel, Starry flounder> Rainbow trout> Israeli mirror carp, Abalone, Whiteleg shrimp. Overall, the future research needs to be done at least more than 3 years of research on the domestic aquaculture farms for the comparison between aquatic medicine use and aquaculture production of each species.
우리나라 양식 흰다리새우(Litopenaeus vannamei)에서 발생한 전염성근괴사증(Infectious Myonecrosis) 역학 조사
권문경(Mun-Gyeong KWON),김수미(Su-Mi KIM),신기원(Ki-Won SHIN),조미영(Mi-Young CHO),황성돈(Seong-Don HWANG),서정수(Jung-Soo SEO),황지연(Jee-Youn HWANG),지보영(Bo-Young JEE) 한국수산해양교육학회 2019 水産海洋敎育硏究 Vol.31 No.1
We carried out an epidemiological survey of infectious myonecrosis which first occurred in farmed whiteleg shrimps in May 2015. The virus was not detected in culture water, feeds, and surrounding organisms in the IMNV-infected farms. In 2015, 5 hatcheries of whiteleg shrimps were operated in Korea, so we carried out virus tests on shrimp broodstock, shrimp feed lug worms(imported), rearing water, and surrounding organisms sampled from each of the 5 hatcheries. The tests detected the virus in one of 6 tested adult shrimps. 25~40% of the imported lug worms were also found infected, which requires us to consider need for import quarantine. The rearing water as well as the 11 species present in the infected hatcheries were also tested positive. These results require us to consider need for further research on correlations between the virus and surrounding animals in infected shrimp farms.
조미영(Miyoung CHO),하헌주(Heon-Ju HA),민진기(Jin-Gi MIN),김태진(Tae-Jin KIM),지보영(Bo-Young JEE),박신후(Shin-Hoo PARK),황성돈(Seong-Don HWANG),김광일(Kwang-il KIM),장영환(Yeounghwan JANG),박명애(Myou]ng-Ae PARK) 한국수산해양교육학회 2015 水産海洋敎育硏究 Vol.27 No.6
Korean government has issued the ratings standard for disease control of aquaculture farms according to the Aquatic Life Disease Control Act in 2008. There are five rating criteria within the enforcement regulations; period of disease free, performance of disinfection, environmental and hygienic management, keeping record of trade and sales, completion of education for disease control. There are no application has been applied on the basis of the rating criteria so far because detailed subcriteria and scoring indicators has not established for assessment process. In order to apply the ratings standard for disease control, we set up the subcriteria and new scoring for rating disease control level based on legal criteria in this study. This study was conducted on 90 targeted olive flounder farms in Jeju. The result of disease control level assessment on olive flounder aquaculture farms show the overall average of Jeju-do was 72.9 points and differed depending on the district; 74.7 in Jeju-si and 71.1 in Seogwipo-si, respectively. The Spearman correlation of each evaluation index show the disease control level score was correlated highly with the performance of disinfection(R=0.715, p<0.01), and environmental and hygienic management(R=0.661, p<0.01). The result of correlation analysis show that there is the highest correlation between the aquaculture farm status and the total mortality(R= -0.618, p<0.01). The reliability of the results are considered to be accurate in the Cronbach’s alpha value of 0.727.