碩士 / 輔仁大學 / 生物學研究所 / 85 / The immunity system of Penaeus will be activated to defend itself from foreign pathogen attacks. In this study, we used a common shrimp pathogen, Vibrio damsela, as an inducer, and infected shrimp by intramuscular injection and immersion. We injected P. japonicus with Vibrio damsela in different concentrations intramuscularly and determined the LD50 in 5 days. Then we injected P. japonicus with Vibrio damsela in this dose, and took the plasma from the hemolymph for 10% native-PAGE analysis. The result showed that an extra protein fragment in low-molecular weight appeared at 6 hours post-infection. The peak of this protein production was observed at 8 hours post-infection. At 12 hours post-infection, the protein declined and disappeared after 24 hours.
In addition, we immersed P. japonicus in Vibrio damsela containing sea water 3 hours and applied the plate count method to determine the bacteria clearance capability of the shrimp hemolymph withdrawn at various time points. The result showed that the amount of bacteria in the blood declined significantly at 6 hours post-infection, and declined more remarkable at 12 hours post-infection. Twenty four hours later, there was no much difference between the test and the control groups. From this result, we suggested that there were some bactericidal substances produced by the Vibrio damsela pretreated shrimp and these materials could last a period. We applied the PCR-Select cDNA Subtraction Kit, developed by CLONTECH, to figure out if there was difference in mRNA between the 6-hour Vibrio damsela immersed shrimp and the clean water immersed control. As a result, three extra cDNA fragments were found in the subtracted immersed shrimp sample and their sizes were 1.0kb, 0.6kb and 0.5kb, respectively. However, there was no significant difference in mRNA between the test group and the control group as analyzed by the Northern blotting hybridization. To compare the nucleic acid sequences of these three fragments with the published sequences in the GCG gene bank, no high enough homology was found.
We also treated the P. monodon lymphoid cells, PMO (Penaeus monodon oka) cells, with 0.2 μg/μl of glucan. After 1-hour glucan treatment cells became rounded, shrunk and aggregated. These phenomena were more remarkable with time. After 24-hour treatment, lysis of cells was observed.
| Identifer | oai:union.ndltd.org:TW/085FJU03112002 |
| Date | January 1997 |
| Creators | 郭晉祿 |
| Contributors | 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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