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Detection of Ostreid Herpesvirus 1 from adult Pacific Oysters Crassostrea gigas Cultured in Korea
지보영,이수진,조미영,이순정,김진우,최승혁,정현도,김기홍 한국수산과학회 2013 Fisheries and Aquatic Sciences Vol.16 No.2
The presence of ostreid herpesvirus 1 (OsHV-1) and the percentage of viral DNA detected in Pacific oyster Crassostrea gigas adults were investigated monthly between May and November 2012 at three locations along the southern coast of Korea. Among 210 oysters examined by polymerase chain reaction (PCR) analysis, OsHV-1 DNA was detected in only one oyster collected in August. The low detection rate of OsHV-1 DNA was consistent with the lack of reported OsHV-1-associated disease in C. gigas cultured in Korea. The sequence of the present PCR product amplified with the C2/C6 primer pair was identical to that of OsHV-1 μVar except for one nucleotide, and the sequence amplified with Del36-37F2/Del36-37R showed a 605-bp deletion as in OsHV-1 μVar. Although these sequence data are insufficient to determine genotype, the results suggest that the herpesvirus detected was similar to OsHV-1 μVar. This is the first report on the presence of OsHV-1 in adult Pacific oysters cultured in Korea.
수온별 저산소 노출에 따른 조피볼락 (Sebastes schlgeli)의 혈액성상 변화
지보영,도용현,민병화,박미선,황형규,명정인,조재권 한국환경생물학회 2015 환경생물 : 환경생물학회지 Vol.33 No.4
The objective of this study was to evaluate the effects of acute hypoxia on the physiological stress responses (plasma cortisol as the primary response, and hematocrit, hemoglobin, plasma glucose, sodium, chloride, osmolality, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) as the secondary responses) of Korean rockfish. The mortality in Korean rockfish started at 0.7 mg L-1 and 0.8 mg L-1 levels at 12℃ and 20℃, respectively. And the time required for the mortality in experimental fish was 274 and 148 minutes at 12℃ and 20℃, respectively. Levels of hematocrit, hemoglobin, AST, ALT, ion concentration, osmolality, glucose and cortisol had significantly increased with decreasing dissolved oxygen at 12℃ and 20℃.
담수산 백점충(Ichthyophthirius multifiliis)에 관한 연구 II. 백점충의 인위 감염 및 어체내 충체 발달상
지보영,김기홍,박수일,김이청,Ji, Bo-Young,Kim, Ki-Hong,Park, Soo-Il,Kim, Yi-Cheong 한국어병학회 1998 한국어병학회지 Vol.11 No.1
본 연구는 내수면 양식 어류에 백점충, Ichthyophthirius multifiliis를 인위 감염시켜 감염 특성 및 어체내 충체 발달상을 밝히고자 하였다. 어종별(무지개 송어, 메기, 이스라엘 잉어), 수온별($15^{\circ}C$, $18^{\circ}C$, $21^{\circ}C$) 및 충체 발달 단계별(trophont, protomont, theront) 인위 감염 시험 결과, 어종별로는 무지개 송어 시험구에서, 수온별로는 $18^{\circ}C$ 시험구에서 그리고 충체 발달 단계별로는 theront 시험구에서 양성적인 인위 감염을 확인할 수 있었다. 그리고 theront 농도별에 있어서는 어체당 1000개체의 theront가 감염을 양성적으로 유도할 수 있는 것으로 조사되었으며 어체당 1500개체 이상의 theront는 무지개 송어치어에 100%의 감염을 유도하였다. 수온 $18^{\circ}C$의 인위 감염된 무지개 송어에서 7일 동안 어체내 충체의 발달 과정을 조사한 결과, 충체는 시간이 경과함에 따라 연속적인 발달을 행하여 크기가 증대되었으며 감염 3일째부터는 성숙되는 것으로 확인되었다. 그리고 총체의 형태학적 변화는 감염 2일째에는 구부장치가 기능적으로 발달하기 시작한 것을 볼 수 있었고, 감염 4일째부터는 수축포가 현저히 발달되기 시작하였으며 감염 6일째는 이들의 수가 현저히 증가한 것을 확인할 수 있었다. 그리고 감염 기간이 경과함에 따라 충체는 숙주의 아가미 일차 새변의 입새 동맥쪽으로 이동하는 경향을 나타내었다. Concerned to the lyfe cycle of Ichthyophthirius multifiliis, the experimental infection and development of the parasites were studied in the several freshwater cultured fishes. Opitimum conditions for the propagation of the parasite by serial passage with the rainbow trout fry was observed. Visiable white spots were examined in the body surface, fins and gills of the healthy fries, and a stable infection has been maintained for 2 months in the experimental system (Temperature: $18{\pm}1^{\circ}C$ DO: 7-7.5 ppm; pH: $7{\pm}0.2$). Induction conditions for artificial infection of the parasite by interms of the host fishes, stages of the parasites, and rearing temperature regimes was investigated. Rainbow trout fries showed a positive infection which was resulted from exposure of theront at $18^{\circ}C$. The rainbow trout fries induced white spots on the body surface at 3-7 days exposure to the theronts at $18^{\circ}C$. It was found that exposure of the rainbow trout fries exposed to 1,000 theronts per fish (10 theront/ml) for 45-60 minutes at $18^{\circ}C$ would consistently produce infection. Perfect infection (100%) was induced when the fries were exposed to 1,500 theront per fish (15 theront/ml) under laboratory condition. Development of I. multifiliis in the rainbow trout was observed for 7 days postexposure (PE). The parasite increased in average diameter from $54{\mu}m$ on the 1st day PE to $426{\mu}m$ on the 7th day PE. In the initial infestation period, the parasites were found on the gill epithelium, and on the 3rd day PE they invaded into the basal part of the gill filament adjacent to the major blood vessels, particularly the afferent vessels. Morphological change of buccal apparatus were observed on the 2nd day PE. Contractile vacuoles were more prominent on the 4th day PE, and they had notable changes on the 7th day PE.
지보영,김민선,조미영,이순정,박명애,김진우,최승혁,정현도,김기홍 한국수산과학회 2014 Fisheries and Aquatic Sciences Vol.17 No.3
RNA interference (RNAi)-mediated transcriptional knock-down of Crassostrea gigas big defensin 1 and 2 genes (Cg-BigDef1 and Cg-BigDef2) was investigated. The cDNA sequences of Cg-BigDef1 and Cg-BigDef2 were identical, excluding an additional fragment of 20 nucleotides in Cg-BigDef1; thus, a long double-stranded RNA (dsRNA) targeting the mRNA of Cg-BigDef2 effectively downregulated both Cg-BigDef2 and Cg-BigDef1. In addition, long dsRNA targeting green fluorescent protein (GFP) did not affect transcription of the two big defensin genes. These results suggest that the transcriptional downregulation of Cg-BigDef1 and Cg-BigDef2 was mediated by sequence-specific RNA interference (RNAi). Despite injection of long dsRNA targeting Cg-BigDef2 into only the adductor muscle, knock-down of Cg-BigDef1 and Cg-BigDef2 was observed in the adductor muscle, hemocytes, mantle, and gills, suggestive of systemic spread of RNAi in C. gigas. Furthermore, the inhibitory effect of dsRNA persisted until 72 h post-injection, indicative of a long-lasting RNAi-mediated knock-down of target genes.
Negatively Charged Membrane을 이용한 해수 중어류질병바이러스의 검출
지보영 ( Bo Young Jee ),김광일 ( Kwang Li Kim ),이순정 ( Soon Jeong Lee ),김기홍 ( Ki Hong Kim ),진지웅 ( Ji Woong Jin ),정현도 ( Hyun Do Jeong ) 한국수산과학회(구 한국수산학회) 2013 한국수산과학회지 Vol.46 No.1
After an outbreak of viral disease in an aquafarm, release of virus(es)from infected fish into environmental seawa- ter has been suspected. ln the present study, we utilized a negatively charged membrane(HAtype)as an efficient method for concentration and detection of fish pathogonic viruses, specally, megalocytivinus and viral hemor- rhagic septicemia virus(VHSV)prent in field-collected seawater samples of inoculated into seawater artificially. Positivly charged viruses adsorbed onto the negatively charged membrane and were cluted with l mM NaOH (pH10.5)following rinsing with 0.5 mM H2SO4(pH3.0). Megalocytivirus and VHSV particles isolated using an- egatively changed HA membrane from seawater inoculated with each virus at a concentration of 10 viral paricles/ mL were of sufficient quantity to show positive results in atwo-step PCR(or RTtwo-step PCR); however, despite it being negatively charged, a cellulose acetata(CA)membraneshowed negative results. ln quantitative PCR, the detection limits of the HA membrane for megalocytivirus and VHSV in seawater were 1.20E+00 viral particles/ mL and 1.22E+01 Viralpartieles/mL, respectivly. The calculated mean recovery yields fiom 1 L seawater spiked with kmown concentrations of megalocytivirus and VHSV particles were 28.11% and 23.00%, respectively. The concentrate of a 1-L samplc of culturing seawater from the aqutank of flounder suffering from VHSV showed clear positive results in PCR when isolated with an HA, but not a CA, membrane, viral isolation using an HA membrane is a practical and reliable method for detection of fish pathogenic virtes in seawater.