碩士 / 國立高雄大學 / 生物科技研究所 / 94 / The present study attempts to investigate the characteristics of the iridovirus that infect king grouper（Epinephelus lanceolatus）in pond culture and than to develope a molecular technique that could detect the virus sensitively. The diseased fish were collected during October, 2005 to March, 2006 from growth the ponds in southern coastal areas of the Taiwan. The virus infection was proved by PCR methods using a pair of primer that has been demonstrated RSIV specific. The results showed those of diseased fish including king grouper、goldlined seabream（Rhabdosargus sarba）、giant seaperch（Lates calcarifer）、crimson snapper（Lutjanus erythropterus ）and common slipmouth（Leiognathus equulus）were infected by RSIV. The virus infected cells showed distend and basophilic, and can be seen in the spleen, kidney, gill, heart and liver. Pathogenicity of the virus was proved by infect the juvenile king grouper, the juvenile of king grouper about 5-9g was using intra-peritoneal injection method with a solution 20×diluted virus extract. The mortality of infected fish was about 90% in 9 days, post infection. The PCR product by using a P3/P4 IRB5 primer pair was sequenced and analyzed to construct the phylogenetic relationships with other iridovirus. The result indicated the king grouper iridovirus（KGIV）may belong to the genus Tropivirus. DIG-labelled DNA probes prepared by PCR method. By dot blot hybridization, that were and used to detect the the viral DNA. The probe was proved can detect the viral DNA at a level of 1pg/μl. The probe is sensitive to detect the virus infected fish though some false positive may happen. Using the iridovirus specific primer to establish the PCR diagnosis showed that primers set ATPase 3 and P3 IRB5/P4 IRB5 are more suitable for detection of KGIV. Quantitative analysis the viral genome in various tissue of the infected fish by real-time PCR, the spleen and kidney that were found 1010 viral genome copies/g tissue where the major organs the virus proliferate. The minor organs such as gill, heart and gill also have 109 viral genome copies/g tissue. We suggest that PCR method is more suitable for detecting the king grouper whether suffer from iridovirus. By these methods that can examination of the king grouper whether suffers from iridovirus infection.
| Identifer | oai:union.ndltd.org:TW/094NUK05111005 |
| Date | January 2006 |
| Creators | Jui-Cheng Chang, 張睿城 |
| Contributors | Chun-Shun Wang, 王俊順 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 76 |
Page generated in 0.007 seconds