碩士 / 輔仁大學 / 生命科學系碩士班 / 97 / Iridovirus infection has been damaging devastatingly on Taiwan aquaculture in these few years. The fish species susceptible to iridovirus including giant seaperch Lates calcarifer, king grouper Epinephelus lanceolatus, Tiger grouper Mycteroperca tigris, Orange-spotted grouper Epinephelus coioides, Cobia; Rachycentron canadum and Red sea bream Pagrus major. The high density aquaculture manner not only causes the abrasion but also increases the tension and pressure of fish. And the insignificant symptom of viral diseases further speed the occurrence and spread of infectious diseases. Family Iridoviridae includes five genera: Ranavirus, Lymphocystivirus, Megalocytivirus, Iridovirus and Chloriridovirus. In which, three genera can infect fish: GIV belongs to genus Ranavirus, LCDV belongs to genus Lymphocystivirus and TGIV belongs to genus Megalocytivirus. Grouper iridovirus is a dsDNA virus, icosahedral, 170±10 nm in diameter and with envelop. Diseased fish will lack of appetite, dark color in body, pale color in gill filament, and hemorrhage in gill cover, lower jaw, around eyes, underbelly and gill base. Anatomy observation includes enlargement of spleen and kidney. Iridovirus can infect fish from fry throughout adult fish, and the mortality can reach 50%~100%. Cell line is an important tool for virus isolation and research. In this study, we used fertilized eggs of Orange-spotted grouper Epinephelus coioides as material and established iridovirus susceptible embryonic cell line. The morphology of grouper embryo, GE cell line was epidermal type after subcloning, and was kept in L-15 medium supplemented with 5-20% FBS at 28 ℃. The chromosome number distributed major in 28 and 32. Now the GE cell line has been passing for 40 generation. GE cell line has high susceptibility to grouper iridovirus, and the infection titer can reach 108.5 TCID50/ml. We also transfected pEGFP-N1, pEGFP-N/GIV-Bcl and pDsRed2-Mito into GE cells and found the expression of fluorescent protein. All these results suggest that the grouper embryo (GE) cell line we established is suitable for the cell biology study of iridovirus.
| Identifer | oai:union.ndltd.org:TW/097FJU00105010 |
| Date | January 2009 |
| Creators | CHEN YU-LIANG, 陳昱良 |
| Contributors | CHANG CHI-YAO, 張繼堯 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 70 |
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