摘要: |
于1993-1995年,用免疫法防治皱纹盘鲍脓疱病。河流弧菌-Ⅱ分离自大连水产养殖公司、太平洋海珍品养殖公司的患脓疱病的皱纹盘鲍;健康皱纹盘鲍,由大连水产养殖公司石庙苗种厂提供。鲍血淋巴细胞的体外吞噬实验:取健康皱纹盘鲍的血淋巴与肝素和菌液按一定比例混合后,置湿盒内37°C培养30min,取一滴涂片,吕氏液染色后观察。用该病原菌制做菌苗,大量扩增病原菌,用0.1%-1.0%的甲醛洗下菌苔,处理24-120h,用生理盐水反复清洗,除掉甲醛;再用生理盐水稀释到所需浓度。通过反复传代降低毒性,制成活菌苗。用两种菌苗在稚鲍、一龄鲍(2-3cm)和成鲍(6-8cm)上实验。结果表明,鲍的血淋巴细胞约有三种,三种细胞都有吞噬细菌的能力,有的细胞象变形虫样变形,在细胞膜上吸附着很多细菌,有的细菌已被吞到细胞内;该病原菌可同鲍的血清发生凝集反应;菌苗的效果明显,可提高成活率约50%以上。由上述结果可见,皱纹盘鲍的血淋巴细胞具有吞噬细菌的能力,在菌苗的作用下,血淋巴细胞增殖并活跃地吞噬异源致病菌,同时鲍血液中的免疫因子也被激活,参与免疫的过程,使皱纹盘鲍成活率明显提高。 |
关键词: 皱纹盘鲍 免疫学 河流弧菌-Ⅱ 脓疱病 |
DOI: |
分类号: |
基金项目:大连市科委重大资助项目,1994年44号 |
附件 |
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IMMUNOLOGICAL STUDIES ON HALIOHS DISCUS HANNAI WITH VIBRIO FLUVIALIS-II |
Li Taiwu1, Ding Mingiin2, Xiang Jianhai3, Liu Ruiyu3
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1.Department of Biology,Liaoning Normal University,Dalian 116029;2.Aquacultural Company of Dalian,Dalian 116023;3.Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071
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Abstract: |
Various anti-fibrosis vaccines for cultured abalone were examined in these studies on the immune response of abalone Haliotzs discus hannai against Vibrio fluvialis-II by phagocytosis in bitro, agglutination reaction. Experimental abalones were obtained in 1993 from the farm of the Aquacultural Company of Dalian. Vibrio fluvialis-II strain was used to prepare immunogen and reactive antigen The strain was originally isolated from the diseased abalone supplied by the Aquacultural Company of Dalian and Aquacultural Company of the Pacific Ocean from 1993 to 1995. The strain was incubated at 37°C for l2-16h in medium of sea water and tryptone (SWT) supplemented with 2% NaCl. Hemolymph (40μl) from (an aperture made by a sterile needle in the abalone) heart was mixed with 20μl heparin and 20μl Vibrio fluvialis-II, incubated in a moist box at 37°C for 35min, and then stained for taking photographs of the agglutination reaction, the diluted to twice the volume Vibrio fluvialis-II solution was put into 12 glass tubes respectively. A 13th tube held physiological saline (2% NaCl) solution for producing the interface between the abalone serum and the bacteria solution. The tube in which the interface produced a white precipitation ring showed agglutination reaction. The vaccine (the above-obtained Vibrio fluvialis-II) washed down from the medium with the use of 0.1% - 1.0% formalin for 24 - l20h was centrifuged at 3000-4000 r/min for 5min, then washed another 2 - 3 times with physiological saline (PS) and centrifuged as above. The precipitated bacteria were suspended in the PS at a concentration of 104 - 1011cell/ml. The results showed alter oral ingestion (0.3-6 × 108cell/g) or injection (6.4-19.4 × 108cell/each abalone) of the vaccine, the survival rate of the juvenile, one-year-old, and adult abalone could reach to 70%, 80% and 50% respectively. The abalone had three kinds of leucocytes, or cells phagocytototic to Vibrio fluvialis-II. The serum of the abalone have more agglutinin (1/16) alter the abalone was injected with vaccine made from Vibrio fluvialis-II. |
Key words: Haliotis discus hannai, Immunology, Vibrio fluvialis-II, Pustule disease |