| Summary: | 碩士 === 國立臺灣海洋大學 === 水產養殖學系 === 100 === The transgenic yeast strain LZ-8 was added in feed which according to different protein contents (1.25, 6.25, 12.5 and 37.5 mg LZ-8 protein/kg) feeding grouper (Epinephelus coioides). After fed with feed contains LZ-8 for 1, 3, 5, 7 and 14 days, groupers were injected with NNV and the survival rate was recorded during the experiment. 9 orange-spotted groupers were sampled after 14 days. The results of NNV challenge experiment, 37.5 mg LZ-8 feed fed for 1 day had the best survival rate (83.5%) after 14 days. The increase of feeding days the survival rate was decreased to 58.5 %.
5 different groups of LZ-8 and 2 groups of β-glucan from different materials of collecting (freeze-dried collection and baking collection) and different yeast (Pichia pastoris and Saccharomyces cerevisiae) produced and then extracted from the production. The extraction was used to observe the effect of the in-vitro of non-specific immune in white blood cells for orange-spotted groupers. Results showed that freeze collection produced in Pichia pastoris LZ-8 (1) and β-glucan (c brand) at 0.125 mg/ml was significantly increased in the white blood cell superoxide anion, reactive oxygen species, phagocytosis rate.
The feed which added with LZ-8 (1), β-glucan (c brand) were the treatment groups; whereas LZ-8 (4) and β-glucan (t brand) were acted as control groups. Orange-spotted groupers were fed with all of the treatment groups and the control groups for 1, 2, 4, 7 and 14 days. 9 groupers was taken from each groups and injected with NNV challenge meanwhile the feed which contains LZ-8 and β-glucan was stop to feed the groupers. Survival rate was then recorded for 14 days. The results of challenge experiment, the grouper fed with LZ-8 (1) feed for 1 day had the best survival rate (100%) after 14 days. However, The increase of feeding days the survival rate was decreased to 58.5 %. Conversely, groupers which fed with β-glucan (c brand) had the best survival rate (67 %) but decreased to 33 % after 14 days. Therefore, LZ-8 (1) and β-glucan (c brand) increased the non-specific immune in white blood cells and the resistance of NNV in orange-spotted groupers was also increased.
Besides, NNV anti-monoclonal antibody was used to investigate the effect on the resistance of NNV. Orange-spotted groupers were bath with NNV anti-monoclonal antibody for 0.5 hour and then challenge with NNV bath for 2.5 hours. The experiment was observed for 14 days survival rate was recorded. Results showed that orange-spotted groupers which bath with NNV anti-monoclonal antibody dilution at 1,000~50,000 times increased the resistance of orange-spotted groupers from NNV challenge.
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