【作者】 郭文婷；

【导师】 李健； 唐学玺；

【作者基本信息】 中国海洋大学 ， 海洋生物学， 2005， 硕士

【摘要】 从健康的鱼虾体表体内和养殖水体中分离得到的具有较强拮抗鳗弧菌、副溶血弧菌、沙蚕弧菌、飘浮弧菌等病原性弧菌作用,同时具有蛋白酶、淀粉酶、脂肪酶等酶活力的4株有益细菌,组成5种微生态制剂。以牙鲆为试验动物,在基础饲料中分别添加上述5种微生态制剂N₃、E₅、N₃+E₅,、枯草芽孢杆菌（CICC10073）、N₃+E₅+枯草芽孢杆菌+纳豆芽孢杆菌+乳杆菌,对照组只饲喂基础饲料。在第10、20、30、40、50、60d分别取血,并测定血细胞吞噬活性及血清中的抗菌活力（Ua）、溶菌活力（U1）、超氧化物歧化酶（SOD）活力。结果表明,添加0.4%N₃+E₅微生态制剂对吞噬活性促进作用最为显著;添加N₃微生态制剂对血清中抗菌活力促进作用最为显著;5种微生态制剂对溶菌活力的促进作用尤为显著,第40d,添加了N₃、E₅、N₃+E₅和枯草芽孢杆菌组则分别比对照组提高了195.2%、222.7%、153.0%和113.6%;此外,5种微生态制剂对SOD酶活力也有一定的促进作用。60d后,用鳗弧菌通过腹腔注射进行攻毒试验,试验组的免疫保护率分别达到43.4%、71.6%、40%、62.2%、20%,表明所用微生态制剂可以有效地促进牙鲆的非特异性免疫功能。 在前次实验的基础上,在基础饲料中分别添加N₃,E₅,N₃+E₅等3种微生态制剂,投喂凡纳滨对虾,同时监测对虾血清中酚氧化酶、抗菌活力、过氧化物酶活力及蛋白含量的变化情况,并与一种市售花粉免疫增强剂相对照,评价它们的应用效果。酚氧化酶测定结果表明,试验组较对照组都达到了极显著差异（P<0.01）。3种微生态制剂和花粉免疫增强剂对过氧化物酶也有显著的促进作用（P<0.01）,E₅组和N₃+E₅组高活力值维持到了第25d。抗菌活力的测定结果表明,第7d,E₅组较对照组达到了极显著差异（P<0.01）;第12d,4个试验组全部达到了极显著性差异（P<0.01）;E₅,N₃+E₅组对抗菌活力促进作用最为显著。对血清中蛋白含量定量结果表明,试验组血清蛋白含量也有所提高。从对免疫因子的促进效果来看,E₅和N₃+E₅在对几种免疫因子的促进作用上优于枯草芽更多还原

【Abstract】 We select 4 marine bacterial strains isolated from healthy fish ,shrimp and aquatic environment which have high inhibitory activities to four pathogenic indicating vibrios. Vibrio anguillarum, Vibrio parahaemolyticusi, Vibrio campbelli and Vibrio natriegen, at the same time, they have high digestive activities. Paralichthys olivaceus were fed with basal diet supplemented 0.4% N₃, E₅ , N₃+E₅, Bacillus subtilis （CICC 10073） , N₃+E₅+ Bacillus subtilis +Bacillus natto+ Lactobacillus respectively for two months. Control group was fed with basal diet. The phagocytic activity of blood cells and antibacterial activity,lysozyme activity, superoxide sismutase in the serum were determined. The phagocytic activity fed with dietary N₃+E₅ were changed remarkably on 40th day, the phagocytic rate and phagocytic index were 60.0% and 18.5% higher respectively than those of the control group. The antibacterial activity fed with dietary N3 were 65.7% higher than those of the control group on 40th day. The lysozyme activity fed with dietary N₃, E₅, N₃+E₅, Bacillus subtilis were 195.2%, 222.7%,153.0%,113.6% higher respectively than those of the control group. Moreover, these probiotics also had some effects on the SOD activity. Compared to dietary N₃, E₅, N₃+E₅, Bacillus subtilis groups, the final group were not changed the activity remarkably. After 60 days,fish in each groups were challenged with V. anguillarum intraperitoneal injection, the relative percent survival（RPS） value of the test groups reached 43.4%, 71.6%, 40%, 62.2% and 20%, respectively. The above study results indicated that the different probiotics we used could promote non-special immune competence of Paralichthys olivaceus and showed different effects.Litopenaeus vannamei Boone were fed with basal diet supplemented 0.4% E₅ , N₃+E₅, Bacillus subtilis （CICC 10073） , pollen immunostimulant. Phenoloxidase,peroxidase, antibacterial activity and protein concentration in the serum were detected. The phenoloxidase activity of the test group were all better than the control group（P<0.01）. There are the same effect on the peroxidase activity, the high activity of E5 and N3+E5 were kept through 25d.All of the text group were changed the antibacterial activity remarkably. The effect on protein concentration in the serum of the test group were changed through the test. Accroding to these results, we can draw the conlusion that E5,N3+Es were better than the Bacillus subtilis and pollen immunostimulant.Litopenaeus vannamei Boone were fed by basic diets containing 0, 0.5%, 1%, 2%（M/M） Chinese Herbal Medicine respectively. Phenoloxidase, peroxidase, antibacterial activity and protein concentration in the serum were detected.. All the indexes of the test groups measured were significantly higher （P<0.01 ） than the control group. A challenging test was conducted by injecting the shrimp with Vibrio parahaemolyticus, the mortalities were recorded in 8 days. The relative percent survival（RPS） value of the test groups reached 50 % ⁶5 %, especially in the shrimp fed with diets containing 2% Chinese Herbal Medicine. Accroding to these results, we can draw the conlusion that 1 % ² % is recommended for enhance the immunity and the growth of L. vannamei.更多还原