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Differentiation of Lymphocystis Disease Virus Genotype by Multiplex PCR
키타무라신이치,정성주,오명주 한국미생물학회 2006 The journal of microbiology Vol.44 No.2
Lymphocystis disease virus (LCDV) is the causative agent of lymphocystis disease. The viruses have been divided into three genotypes (genotype I for LCDV-1, II for Japanese flounder isolates, and III for rockfish isolates) on the basis of major capsid protein (MCP) gene sequences. In this study, we developed a multiplex PCR primer set in order to distinguish these genotypes. We also analyzed the MCP gene of a new LCDV isolate from the sea bass (SB98Yosu). Comparison of sequence identities between SB98Yosu and eight Japanese flounder isolates, revealed identity of more than 90.1% at nucleotide level and 96.5% at deduced amino acid level, respectively. Phylogenetic analyses based on the MCP gene showed that SB98Yosu belongs to genotype II, along with Japanese flounder isolates. Multiplex PCR based on the MCP gene allowed us to identify these genotypes in a simple and rapid manner, even in a sample that contained two genotypes, in this case genotypes II and III.
Genetic Variation and Geographic Distribution of Megalocytiviruses
송준영,키타무라신이치,정성주,Toshiaki Miyadai,Shinji Tanaka,Yutaka Fukuda,김석렬,오명주 한국미생물학회 2008 The journal of microbiology Vol.46 No.1
Viruses belonging to the genus Megalocytivirus in the family Iridoviridae have caused mass mortalities in marine and freshwater fish in Asian countries. In this study, partial major capsid protein (MCP) gene of seven Japanese and six Korean megalocytiviruses was sequenced and compared with the known megalocytiviruses to evaluate genetic variation and geographic distribution of the viruses. Comparison of MCP gene nucleotide sequences revealed sequence identity of 92.8% or greater among these 48 isolates. A phylogenetic tree clearly revealed three clusters: genotype I including nine Japanese isolates, thirteen Korean isolates, one Chinese isolates, one Thailand isolate and one South China Sea isolate; genotype II including five freshwater fish isolates in Southeast Asian countries and Australia; and the remaining genotype III mainly consisted of flatfish isolate in Korea and China. This suggests that viruses belonging to the genotype I widely distribute among various fish species in many Asian countries. Conversely, the epidemic viruses belonged to genotype II and III are may be still locally spreading and constrained in their prevalence to the limited host fish species, i.e., genotype II viruses mainly distribute in Southeast Asian countries, whereas genotype III viruses distribute in flatfish species in Korea and China.
김석렬,박명애,키타무라신이치,정성주,오명주 한국수산과학회 2010 Fisheries and Aquatic Sciences Vol.13 No.2
In recent years, several southern coastal fish farms in Korea have experienced 2-30% mortality in striped beakperch suffering from bacterial infections during the spring. In this study, we identified a bacterium isolated from diseased striped beakperch as Pseudomonas anguilliseptica via a biochemical test and 16S rDNA sequence analysis. To evaluate the susceptibility of striped beakperch to P. anguilliseptica, 4.39×107 or 4.39×105 CFU/fish of bacteria were injected intraperitoneally at 18±1°C into fish weighing 5.5 g. Cumulative mortalities reached 100% and 45% in the 4.39×107 and 4.39×105 CFU/fish infected groups, respectively. Experimentally infected fish showed cell-associated inflammation as well as bacteria in the kidneys and spleen. This study is the first report of striped beakperch mortality caused by P. anguilliseptica, which has pathogenicity in striped beakperch.
Manami Inaba,Satoru Suzuki,키타무라신이치,Norichika Kumazawa,Hiroshi Kodama 한국미생물학회 2009 The journal of microbiology Vol.47 No.1
Distribution of marine type of Aquabirnavirus (MABV) was examined in shellfish and fish from Okinawa and Ishigaki Islands, Japan, where water temperature is higher than 25°C through the year. Genome detection and virus isolation were performed for shellfish and fish samples, and the results revealed the prevalent distribution of MABV in diverse species in the area, although isolation was not frequently. Detection rate of MABV genome in bivalves was higher than gastropods, which was similar result to former report in mainland of Japan. Furthermore, the unique five-nucleotide deletion was found with a high rate of occurrence in the MABV genome from shellfish and fish. This study showed distribution status of MABV in organisms in subtropical waters by wide monitoring, and discovered new genome variation in VP2/NS region of this virus.
양식산 Tubot, Scophthalmus maximus 감염원으로 확인된 Iridovirus의 분자생물학적 연구
김기홍,김위식,김춘섭,김영진,키타무라신이치,정성주,정태성,오명주 한국양식학회 2004 韓國養殖學會誌 Vol.17 No.1
2003년 6월과 7월, 고창에서 사육중인 turbot이 약 90% 누적폐사되었고, 주 증상은 안구돌출, 비장 비대 그리고 장내 내용물이 없는 것으로 관찰되었다. 병리조직학적 검사에서 RSIV와 유사한 이형비대세포가 모든 장기에서 관찰되어 RSIV검출을 위한 primer로 PCR을 실시하였다. 그 결과 PCR에서 특이밴드가 모든 조직에서 검출되어졌고, 염기서열 분석 결과 기존에 보고된 strain과 88%이상의 유사성을 나타내는 것으로 확인 되어져서 터봇의 페사 원인으로 이리도바이러스감염이 관여됨을 확인할 수 있었다. The high mortality of cultured turbot, Scophthalmus maximus was occurred from Gochang in June, 2003. Infect fish with iridovirus showed a lower feed intake and lethargic. These fish exhibited pale body color, extended abdominal and exophthalmus. Histopathological studies showed basophilic enlarged cells from kidney, spleen, gill, heart, muscle, stomach, intestine, liver and pancreas of these fish. In PCR amplification with red sea bream iridovirus (RSIV) specific primer set of a DNA fragment of 286 bp was obtained from infect turbot with the virus. The strain showed the high homologies with RSIV, LBIV (largemouth bass iridovirus), GSIV (giant seaperch iridovirus) and SBIV (sea bass iridovirus).