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        • KCI등재

          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.

        • KCI등재

          생약제 첨가사료가 넙치의 백신효과에 미치는 영향

          황지연(Jee-Youn HWANG),권문경(Mun-Kyoung KWON),서정수(Jung-Soo SEO),김경덕(Kyoung-Duck KIM),이예숙(Ye-Suk LEE),정승희(Sung-Hee JUNG) 한국수산해양교육학회 2015 水産海洋敎育硏究 Vol.27 No.5

          In this study we investigated the effect of vaccine efficacy and immune activities by feeding additives several natural herbal remedies extracts of Sweet Wormwood (Artemisia annua), Galla Rhois, Oriental raisin tree (Havenia Dulicis) and 6-mixed herb to Oilve flounder, Paralichthys olivaceus. Each group of fish fed with the herbal remedies added feed and basic formula during the seven weeks and vaccinated with Edwardsiella tarda Formalin killed vaccine after 4 weeks feeding. Relative Percentage Survival values (RPS) in the group was assessed by the challenge with E.tarda. All groups with the herbal remedies added feed enhanced growth rate, but there were not significantly different in lysozyme activity and agglutination titer. In a challenge experiment with E.tarda, RPS in the all groups feeding additives natural herbal remedies was higher than that of the control group. These results suggest that the natural herbal remedies extracts of Sweet Wormwood (Artemisia annua), Galla Rhois, Oriental raisin tree (Havenia Dulicis) and 6-mixed herb would be effective to enhance efficacy of vaccination to Olive flounder.

        • KCI등재

          Molecular Characterization of Megalocytiviruses from Diseased Fishes in Korean Aquatic Farms from 2013 to 2017

          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.

        • KCI등재

          In vitro 조건에 따른 molecular adjuvant의 넙치, Paralichthys olivaceus 면역유전자 자극 효과

          권문경(Mun-Gyeong KWON),황지연(Jee-Youn Hwang),서정수(Jung-Soo SEO),정승희(Sung-Hee JUNG) 한국수산해양교육학회 2015 水産海洋敎育硏究 Vol.27 No.5

          Adjuvant is an immune enhancer commonly used during vaccination to enhance the host immune response. In the present study, we produced the several recombinant protein from immune related gene of olive flounder (Paralichthys olivaceus). Especially, to produce the soluble type of recombinant protein, we constructed the MBP (Maltose binding protein) fusion G-CSF (Granulocyte colony stimulating factor) recombinant protein among the flounder immune related genes. To verify the immune stimulatory effect and safety of this recombinant protein (rPoGCSF), expression changes of several immune genes were tested using quantitative real-time PCR method with gene specific primer from flounder head kidney leukocytes. As a result, we confirmed that the rPoGCSF has an ability of immune stimulatory effect, also it has broad range of pH and temperature.

        • KCI등재

          바이러스성 출혈성 패혈증 바이러스 NV 단백질에 의한 glucokinase 전사 활성의 억제

          조미영(Mi Young Cho),황지연(Jee Youn Hwang),지보영(Bo Young Ji),박명애(Myoung Ae Park),성미소(Mi So Seong),김소영(So Young Kim),정예은(Ye Eun Jung),정재훈(Jae Hun Cheong),최영현(Yung Hyun Choi) 한국생명과학회 2016 생명과학회지 Vol.26 No.12

          바이러스성 출혈성 패혈증 바이러스(VHSV)는 넙치를 포함한 어류 양식의 막대한 피해를 일으키는 바이러스 병원체이며, VHSV가 생성하는 6개의 바이러스 단백질들 중에서 NV 단백질이 병원성에 관여하는 것으로 알려져 있다. VHSV-감염 넙치를 이용한 전사체 마이크로 어레이의 선행 분석 결과에 의하면 VHSV 감염이 해당과정 효소들의 mRNA 발현을 억제함으로써 넙치 세포에서 ATP 생성을 감소시켰음을 알 수 있었다. 이들 결과를 토대로, 본 연구에서는 VHSV NV 단백질이 해당과정 효소인 glucokinase의 발현에 미치는 영향을 검토하였다. 본 연구의 결과에 의하면, NV 단백질은 넙치 세포에서 glucokinase의 mRNA 발현을 감소시켰으며, 새롭게 동정한 glucokinase의 유전자 프로모터의 활성 실험결과, NV 단백질이 glucokinase의 프로모터 활성을 저해함을 알 수 있었다. 이와 같은 작용 결과들로 인하여 VHSV NV 단백질의 발현이 세포 내로의 포도당 흡수 또한 감소시켰다. 이러한 결과들은 VHSV NV 단백질이 유전자 발현의 전사 수준에서 음성적으로 해당과정의 효소 발현을 조절함을 의미하며, 결국 세포 내 에너지의 결핍으로 넙치의 폐사로 이어질 가능성을 보여주는 것이다. The viral hemorrhagic septicemia virus (VHSV), which belongs to the Novirhabdovirus genus of the Rhabdoviridae family, is a viral pathogen that causes severe losses in the olive flounder farming industry. Among six encoding VHSV proteins, the non-virion (NV) protein has been shown to have an impact on virulence. In our previous studies, transcriptomics microarray analysis by using VHSV-infected olive flounder showed that VHSV infection significantly down-regulated the mRNA expression of glycolytic enzymes. In addition, VHSV NV protein variants decreased the intracellular ATP level. Based on these results, we have tried to examine the effect of VHSV NV protein on glycolytic enzyme glucokinase expression, which phosphorylates glucose to glucose 6-phosphate. Our results indicated that the NV protein significantly decreased the mRNA expression of glucokinase in olive flounder HINAE cells. Furthermore, the NV protein played a negative role in the promoter activation of glucokinase. Furthermore, glucose uptake was effectively inhibited by VHSV infection and NV protein expression in olive flounder HINAE cells. These results suggest that the VHSV NV protein negatively regulates glycolytic enzyme expression by a transcription level and eventually leads to gradual morbidity of olive flounder through cellular energy deprivation. The present results may be useful for the prevention and diagnosis of VHSV infection in olive flounder.

        • KCI등재

          PCR 기법을 이용한 2009년 우리나라 서해안과 남해안 바지락, Ruditapes philippinarum의 Perkinsus olseni 감염에 관한 보고

          이남실 ( Nam Sil Lee ),황지연 ( Jee Youn Hwang ),최동림 ( Dong Lim Choi ),박명애 ( Myoung Ae Park ) 한국어병학회 2010 한국어병학회지 Vol.23 No.2

          Prevalence of a protozoan parasite Perkinsus olseni in Manila clam Ruditapes philippinarum was surveyed from July to December 2009 on the west and south coast of Korea. P.olseni infection was diagnosed using two primer sets, P.olseni NTS Forward/P.olseni NTS Reverse set and PolsITS-140F/PolsITS-600R set in polymerase chain reaction(PCR). The results using PolsITS-140F and PolsITS-600R primer set was retained up to 60% at all stations from July to December, except for Padori. Especially, Goheung showed 100% prevalence from October to December. The results about comparison of the 4 station`s DNA sequences which were analyzed from PCR products(457bp) using PolsITS-140F and PolsITS-600R primer set, there were only 2base differences at Sunjedo.

        • KCI등재

          넙치, Paralichthys olivaceus에서 Edwardsiella tarda, Streptococcus iniae와 S. parauberis 혼합백신의 효능 및 안전성 연구

          권문경(Mun-Gyeong KWON),황지연(Jee Youn HWANG),정성희(Sung Hee JUNG) 한국수산해양교육학회 2014 水産海洋敎育硏究 Vol.26 No.6

          Edwardsiella tarda, Streptococcus iniae and S. parauberis are main bacterial pathogens in aquaculture farms of olive flounder, Paralichthys olivaceus. We have discussed the efficacy and safety of 3 type-combined vaccines (A: S. iniae 1mg + S. parauberis 1mg + E. tarda 1mg, B: S. iniae 1mg + S. parauberis 1mg + E. tarda 0.5mg, C: S. iniae 1.5mg + S. parauberis 1.5mg + E. tarda 1mg) through intraperitoneal injections in olive flounder. None of the vaccines gave rise to any signigicant side effects on histopathology and blood chemistry. The antibody titers and lysozyme activities of A type were higher than those of B, C and control. Four weeks after vaccination, RPS (relative percent survival rates) was 62.5~75% (A type), 50~66.7% (B type) and 55.6~62.5% (C type) respectively. As the results, the combined vaccines are possible to prevent edwardsiellosis and streptococcosis, and A type : S. iniae 1mg + S. parauberis 1mg + E. tarda 1mg, is the most effective out of them.

        • KCI등재

          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.

        • KCI등재

          넙치 및 조피볼락 면역 활성에 대한 사료첨가제로서 갯지렁이 항균펩타이드의 효과

          권문경(Mun-Gyeong KWON),서정수(Jung Soo SEO),황지연(Hwang Jee YOUN),박찬일(Chan-Il PARK),정지민(Ji-Min JEONG),배진솔(Jin-Sol BAE) 한국수산해양교육학회 2016 水産海洋敎育硏究 Vol.28 No.6

          본 연구에서는 다양한 면역 유도제를 주사하여 갯지렁이의 면역을 유도시킨 후, 갯지렁이의 항균 펩타이드를 가장 많이 발현시킬 수 있는 최적의 면역 유도제를 선정하였으며, 선정된 면역 유도제로 면역을 유도한 갯지렁이를 분말화 하여 사료에 첨가하였을 때 양식 어류의 면역 활성을 증가시킬 수 있는지에 대한 여부를 조사하였다. 넙치와 조피볼락은 면역활성을 위한 갯지렁이 분말의 사료 첨가율이 서로 달랐으며, 넙치의 경우 0.1%의 사료 첨가율이 넙치의 라이소자임 활성, 혈청 내 코르티졸 농도뿐만 아니라 E. tarda에 대한 항병성에 가장 높은 효과를 보였다. 조피볼락은 0.5%의 사료 첨가율이 조피볼락의 라이소자임 활성을 증가시키고 혈청 내 코르티졸 농도를 감소시켰으며 S. iniae에 대한 항병성 또한 증가시켰다. 따라서 넙치와 조피볼락은 각각 면역 유도된 갯지렁이 분말이 0.1%와 0.5% 첨가된 사료를 급이하였을 때 면역 활성을 가장 많이 유도 할 수 있을 것으로 생각되지만, 반면 그 이상의 농도로 면역 유도된 갯지렁이 분말을 사료에 첨가하여 급이할 시 오히려 면역 활성에 부정적인 영향을 미칠 수 있을 것으로 생각된다. 이번 연구에서는 주간 8 갯지렁이 분말 첨가 사료를 급이한 것에 대한 양식 어류의 면역 활성을 검증 하였으며, 추후 연구에서는 갯지렁이 첨가사료의 급이기간을 고려한 연구가 진행되어야 할 것으로 생각된다. In this study, the effect of the polychaete antimicrobial peptide as feed additives on fish, olive flounder (Paralichythys olivaceus) and black rockfish (Sebastes schlegelii), immune activity was described. The antimicrobial peptide of the polychaete was induced by peptidoglycan from Micrococcus luteus. The fish were fed an experimental diet supplemented with 0%, 0.05%, 0.1%, 0.5% or 1% of immune induced the polychaete to a commercial diet. Haematological parameters, nonspecific immunes and stress were evaluated 2, 4, 6 and 8 weeks during fed. The resistance against bacteria, Edwardsiella tarda and Streptococcus iniae, were analysed on after 8 weeks. The haematological parameters were not significantly changed among tested groups. But the lysozyme activities were significantly high in the 0.1% and 0.5% supplement group of olive flounder and black rockfish, respectively. Additionally, cortisol in plasma was low in the 0.1% and 0.5% supplement group of olive flounder and black rockfish, respectively. And resistance of these supplement groups were significantly induced against bacterial injection.

        • KCI등재

          뱀장어(Anguilla japonica)에 대한 Edwardsiella tarda 불활화 백신의 침지 및 경구 투여 효과

          정승희(Sung-Hee JUNG),권문경(Mun-Gyeong KWON),서정수(Jung-Soo SEO),황지연(Jee Youn HWANG) 한국수산해양교육학회 2015 水産海洋敎育硏究 Vol.27 No.3

          본 연구는 뱀장어 양식에서 오랜 기간 문제시되고 있는 에드와드병(Edwardsiellosis)에 대한 효과적인 백신 투여 방법의 가능성을 알아보고자 하였다. 뱀장어 유래의 Edwardsiella tarda로부터 포르말린 불활화백신(FKC)을 제작하여, 크기가 다른 3개 그룹의 뱀장어(26.8±1.2 g, 7.1±0.7 g, 2.2±0.4 g)에 침지 및 경구 투여를 실시하였다. 그리고 혈청 응집항체가의 활성과 E. tarda 공격접종에 대한 방어력(상대생존율, RSP)의 변화를 조사하였다. 모든 그룹에서 응집항체가와 공격접종에 의한 방어력은 상관성이 없었다. 그러나 26.8g±1.2 g 그룹의 경우, 침지(10 mg/mL)와 침지(10 mg/mL)+경구(10 mg/g) 실험구에서 RSP는 각각 62.6% 및 52.2%를 나타내었으며, 7.1±0.7 g 그룹의 경우, 침지(10 mg/mL)+경구(10 mg/g) 실험구에서 RSP는 56.8%로 나타나서 방어력(RPS>50%)이 확인되었다. Edwardsiellosis has become a serious diseases problem in cultured eels for many years. This study was performed to investigate possibility of vaccination against edwardsiellosis caused by Edwardsiella tarda. We conducted a immersion and/or oral vaccination using formalin-killed E. tarda in eel Anguilla japonica. Three groups of fish (26.8±1.2 g, 7.1±0.7 g and 2.2±0.4 g) were used in this study. The protection (relative percentage survival, RPS) and serum antibody response (agglutination titer) were evaluated in the vaccinated fish. No correlation between agglutination titer and survival rate was observed in vaccinated fish. However, there was a satisfactory protective (RPS>50%) in vaccinated fish. Immersion (10 mg/mL, 1 hr) and immersion (10 mg/mL, 1 hr) plus oral (10 mg/g, 10 days) of 26.8±1.2 g, immersion (10 mg/mL, 1 hr) plus oral (10 mg/g, 10 days) of 7.1±0.7 g showed RPS of 62.6%, 52.2% and 56.8%, respectively.

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