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In-situ hybridization 법을 사용한 양식 넙치, Paralichthys olivaceus의 바이러스 감염 질병 특성 고찰
도정완 ( Jeong Wan Do ),이남실 ( Nam Sil Lee ),정승희 ( Sung Hee Jung ),김경길 ( Kyung Kil Kim ),최혜승 ( Hye Sung Choi ),박정우 ( Jeong Woo Park ),김이청 ( Yi Cheong Kim ) 한국어병학회 2013 한국어병학회지 Vol.26 No.3
PCR (polymerase chain reaction) 법은 신속하고 정확하여 바이러스성 질병진단을 위해 널리 사용되지만 조직병리학적인 정보를 제공하지 못한다. 반면에 in-situ hybridization (ISH) 법을 사용하면 바이러스를 빠르게 검출할 수 있을 뿐 아니라 조직에서의 분포도알 수 있다. 본 연구에서는 RSIV, VHSV, 그리고 VNN바이러스들의 조직내 분포 및 조직 병리학적인 특성을 확인하기 위하여 이 바이러스들에 감염된 에 감염된 양식 넙치의 어류의 다양한 조직들을 대상으로 ISH 법을 적용하였다. 그 결과 이들 세 종류의 바이러스가 각각 다른 조직 및 세포들에 감염함을 확인 할 수 있었다. 본 연구 결과는 ISH법이 어류 병원성 바이러스의 신속 검출 뿐 아니라 조직 병리학적인 특성 확인에도 유용함을 제시한다. Polymerase chain reaction (PCR) is the most rapid and widely used method to detect viral pathogens. However, this method does not provide histopathologic nature of the virus. In situ hybridization (ISH) with oligonucleotide probes is attractive because it is a rapid method for detection and identification of viral pathogens at sites of tissue infection. In order to understand the histopathologic characterictics of Red sea bream iridovirus (RSIV), viral-hemorrhagic septicemia (VHS) virus and viral nervous necrosis (VNN) virus to cultured olive flounder, we her applied ISH method to various kinds of olive flounder tissues with PCR-positive for these three viruses. We found that these viruses showed different tissue tropism and were detected from different cell types. Our results suggest that ISH is useful not only in rapid detection of viral pathogens but also in understanding the histopathologic characters of specific viral pathogens.
Aeromonas veronii의 실험적 감염에 의한 양식 메기, Silurus asotus의 병리학적 변화
김진도 ( Jin Do Kim ),도정완 ( Jeong Wan Do ),최혜승 ( Hye Sung Choi ),서정수 ( Jung Soo Seo ),정승희 ( Sung Hee Jung ),조혜인 ( Hyae In Jo ),박명애 ( Myung Ae Park ),이남실 ( Nam Sil Lee ),박성우 ( Sung Woo Park ) 한국환경생물학회 2013 환경생물 : 환경생물학회지 Vol.31 No.4
Recently, a new disease showing symptoms such as epidermal exfoliation and muscular necrosis occurred in cultured Korean catfish. Although the mortality of fishes was low but the economic damages owing to loss of commercial value were severe. The authors isolated the causative agent from diseased fish and observed pathological changes both in naturally and artificially infected fish. The causative bacteria was identified as Aeromonas veronii. Subsequently we observed the daily death and pathological symptoms of artificially infected fish with Aeromonas veronii. Symptoms of artificially infected fish were similar to those of naturally infected fish and all fish died within 7 days after infection. Histopathological changes on the naturally infected fish revealed severe congestion and necrotic degeneration in the liver, spleen and kidney. Some bacterial aggregates with inflammatory degeneration were observed in the heart, and congestion and fibrosis in the lamina propria of digestive tube were predominant. In artificially infected fish, skin erosion and necrotic degeneration of muscle tissue around injected region were particularly manifested. Degeneration of hepatocytes in liver and hyalic degeneration around ellipsoids in spleen were partially observed. However, there were no predominant signs in digestive tube in artificially infected fish.
2010년 한국 연근해 자연산과 양식산 넙치, 참돔, 감성돔, 조피볼락의 병원체 비교
박명애 ( Myoung Ae Park ),도정완 ( Jeung Wan Do ),김명석 ( Myoung Sug Kim ),김석렬 ( Seok Ryel Kim ),권문경 ( Mun Gyeong Kwon ),서정수 ( Jung Soo Seo ),송준영 ( Jun Young Song ),최혜승 ( Hye Sung Choi ) 한국어병학회 2012 한국어병학회지 Vol.25 No.3
This study surveyed for the prevalence of parasites, bacteria and viruses in four fish species, olive flounder (Paralichthys olivaceus), red sea bream (Pagrus major), black sea bream (Acathopagrus schlegeli) and black rockfish (Sebastes schlegeli) in 2010. The survey was aimed to compare the pathogens detected from wild and cultured fish for an epidemiological study. Anisakis sp. was predominantly detected from wild olive flounder and red sea bream (58.6% and 41.7% respectively), but not from the cultured fishes, suggesting anisakid infection is rare in cultured fish. The wild fish get in contact with the anisakids through their prey such as small fishes or crustaceans which carry the anisakids; whereas the cultured fish are fed with formulated feed, free of anisakids. Bacterial detection rates from the wild fishes examined in the study were lower than those of cultured fishes. Vibrio sp. dominated among detected bacterial population in cultured olive flounder (18%). Since vibriosis is known as a secondary infection caused by other stressful factors such as parasitic infections, handling and chemical treatment, it seems that cultured olive flounder are exposed to stressful environment. Viruses diagnosed in the study showed difference in distribution between wild and cultured fishes; hirame rhabdovirus (HRV) (0.1%) and lymphocystis disease virus (LCDV) (3.9%) were detected in the cultured olive flounder, but not in the wild fish, and marine birnavirus (MBV) (1.7%) and red sea bream iridovirus (RSIV) (3.2%) were detected from the wild and cultured red sea bream, respectively. From the survey conducted, it can be concluded that even though some pathogens (Trichodina sp., Microcotyle sp., etc.) are detected from both the wild and cultured fish, pathogens such as Anisakis sp., Vibrio sp. and LCDV showed difference in distribution in the wild and cultured host of same fish species and this can be attributed to their environmental condition and feeding.
2007년∼2011년 하절기에 양식 넙치, 조피볼락, 참돔, 새우의 세균 및 기생충 감염 현황
정승희 ( Sung Hee Jung ),최혜승 ( Hye Sung Choi ),도정완 ( Jeung Wan Do ),김명석 ( Myoung Sug Kim ),권문경 ( Mun Gyeong Kwon ),서정수 ( Jung Soo Seo ),황지연 ( Jee Youn Hwang ),김석렬 ( Seok Ryel Kim ),조영록 ( Yeong Rok Cho ) 한국어병학회 2012 한국어병학회지 Vol.25 No.3
2007년∼2011년 하절기에 국내 동·서·남해안과 제주의 육상양식장, 가두리 및 축제식 양식장에서 사육중인 넙치 (Paralichthys olivaceus), 조피볼락(Sebastes schlegeli), 참돔 (Pagrus major), 흰다리새우(Litopenaeus vannamei) 의 총 2,413마리에 대하여 세균과 기생충의 감염현황을 조사하였다. 조사기간(2007년∼2011년) 사이 어종별 병원체에 대한 감염양상을 종합하면, 기생충과 세균의 단독감염이 혼합감염에 비하여 비교적 높게 나타났으며, 단독감염 가운데 기생충이 세균보다 감염률이 높은 경향을 나타내었다. 조사기간 동안 넙치, 조피볼락, 새우에서 Vibrio spp. (V. harveyi, V. ichthyoenteri, Vibrio sp.)가 가장 우점종으로 검출된 세균의 위치를 차지하였으며, 참돔에서는 Vibrio sp.와 P. damselae가 우점종 세균으로 검출되었다. 조사기간 동안 가장 우점종으로 검출된 기생충은 넙치에서 스쿠티카충 (M. avidus)과 트리코디나충 (Trichodina sp.), 조피볼락에서 아가미흡충 (M. sebastes), 참돔에서 아가미흡충 (M. tai), 새우에서 섬모충 (Zoothamnium sp.) 으로 나타났다. Diagnostic monitoring in fish farms with land-based tanks and netpen cases were conducted in eastern, western, southern and Jeju island of Korea during summer of 2007∼2011. In total, 2413-fish samples of 4 marine fish species were tested for the detection of bacteria and parasite. Fish species tested were olive flounder (Paralichthys olivaceus), black rockfish (Sebastes schlegeli), red sea bream (Pagrus major), pacific white shrimp (Litopenaeus vannamei). During the diagnostic monitoring from 2007 to 2011, the infection rates by single infection of bacterial or parasitic pathogens were relatively higher than the mixed infections. The main bacterial pathogens in olive flounder, black rockfish and pacific white shrimp were Vibrio spp. (V. harveyi, V. ichthyoenteri, Vibrio sp.). The main bacterial pathogens in red sea bream were also Vibrio sp. and Photobacterium damselae subsp. damselae. The main parasitic pathogens were both Miamiensis avidus and Trichodina sp. in olive flounder, Microcotyle sebastes in black rockfish, Microcotyle tai in red sea bream and Zoothamnium sp. in pacific white shrimp.
양식 넙치 치어에서 분리한 birnavirus 의 특성
손상규,박정우,박명애,정초록,도정완 한국어병학회 1995 한국어병학회지 Vol.8 No.2
최근 남해안 일대의 육상 양식장에서 사육중이던 넙치(Paralichthy olivaceus) 치어가 폐사하여 조사한 결과 3개 양식장에서 바이러스가 분리되었다. 분리된 바이러스들은 모두 외막이 없는 정육면체 모양이었으며 50∼55㎜ 정도의 크기를 지녔다. 전기영동상에서 RNA와 구조 단백질의 patterns를 확인하고, IPNV에 대한 항혈청을 사용하여 중화실험을 수행한 결과, 분리한 세 바이러스는 birnavirus인 IPNV와 매우 유사함이 밝혀졌다. 특히 분리 바이러스중 CS는 IPNV의 AB 혈청형과 DS와 YJ는 SP 혈청형과 유사하였다. During 1993 and 1994, some mortalities of flounder(Paralichthy olivaceus) fry were recorded in several fish farms and viruses were isolated from 3 of the farms. Electron microscopic examination revealed that the virus particles were hexagonal and unenveloped with an average diameter of 50 to 55nm. Serological and molecular properties of these isolates were examined. The viral RNA and polypeptides patterns on electrophoresis, as well as neutralization test results, showed that these isolates were birnaviruses and two were closely related to infectious pancreatic necrosis virus(IPNV) serotype AB and one was to IPNV serotype SP. This is the first isolation of birnaviruses from marine fish in Korea