碩士 / 國立臺灣海洋大學 / 水產養殖學系 / 92 / Abstract
Aeromonas hydrophila and Edwardsiella tarda are fish pathogen. It involves a serious financial loss since the fish pound was infected. Businessmen usually use probiotics to control the aquatic environment and antibiotics to prevent disease. We will decrease the damage which pathogen causes and cut down the use of unnecessary antibiotics if we can examine pathogen in water in advance. It also saves medicine cost and protects food safety from antibiotics.
Recent years, the establishment of SSU rRNA oligonucleotide probe hybridization is an essential technique to study microorganism of biomolecular technology. Informations of Aeromonas hydrophila and Edwardsiella tarda were collected from GeneBank/EMBL. That16S rRNA sequences after compared from program of BLAST.The resultant data used to produce specific oligonucleotide probes as S-S-Aero-0460-a-A-20 (AH460), S-S-Aero-0641-b-A-20 (AH641)及S-G-Edwar-0996-a-A-20 (ET996) respectively. In addition, used virulent gene oligonucleotide probes as aerolysin, amylase, lipase, nuclease, protease, Ehly, citC, gadB, katB, fimA, orfA, ssrB and mukF.
To examine the virulent gene expression in organism by challenge test. We choose E. tarda which RNA is not expressed. After challenge test, we separate bacteria from liver of the dead tilapia and extract RNA of the separated bacteria. Hybridization for RNA analysis with virulent gene oligonucleotide probes has shown that the expression of Ehly, gadB and mukF gene is 100%, of citC, katB and fimA gene is 90.01%, of ssrB gene is 36.36%, of orfA gene is 18.18 %. The virulent genes of bacterial strains would be shut down when it is frozen, but turn on by challenge test. Therefore, this disease may be associated with the expression of virulent gene.
In order to investigate different virulent gene existence and expression of A. hydrophila and E. tarda, we use oligonucleotide probes as AH460, AH641, ET996 and virulent gene oligonucleotide probes to hybridize four fresh cultured ponds in different condition. This has resulted in difference in Ehly and implied the relation with pathoge
| Identifer | oai:union.ndltd.org:TW/092NTOU5086014 |
| Date | January 2004 |
| Creators | Ya-Ru Shen, 沈雅茹 |
| Contributors | Jau-Der Chen, 陳昭德 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 120 |
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