【作者】 宋靖芳；

【导师】 黄文芳；

【作者基本信息】 华南师范大学 ， 动物学， 2004， 硕士

【摘要】 从中草药中寻找抑菌活性物质是目前防治细菌性鱼病的有效途径之一。我国中草药资源丰富，取材方便。本文以嗜水气单胞菌ST-3-3(Aeromonas hydrophila ST-3-3)、豚鼠气单胞菌CSS-4-2(Aeromonas caviae CSS-4-2)、鳗弧菌E3-1-1(Vibrio anguillarum E3-1-1)、沙鱼弧菌PHK，PHS(Vibrio carchariae PHK，PHS)等鱼类病原菌为供试菌种，选取了101种中草药，对其抑菌活性进行了较为系统的测定，并对其中抑菌活性较高的五倍子和诃子进行了较为深入的研究，得出以下结论： 通过对101种中草药的抑菌活性的测定，发现有68种中草药对至少一种病原菌有抑制作用，其中有强抑制活性的有48种。以五倍子、诃子和大蒜的抑菌谱较广，对所有供试菌株的抑菌活性最强。五倍子的抑菌环直径最大可达32.84mm，MIC值0.0488mg／ml；诃子的抑菌环直径最大为28.72mm，MIC值为1.18725mg／ml；大蒜的抑菌环最大达28.80mm，MIC值0.78125mg／ml。 单味药五倍子、诃子和各种配比的药物均对鱼类病原菌有强抑制作用。五倍子单用时，MIC值从0.048828mg／ml～0.78125mg／ml，MBC值在0.097656mg／ml～0.78125mg／ml；诃子单用时，抑菌活性较五倍子弱，MIC值从0.097656mg／ml～6.25mg／ml，MBC值从0.195313mg／ml～25mg／ml；五倍子和诃子对5株病原菌的抑菌活性强弱分别是：PHK＞PHS＞CSS-4-2＞ST-3-3＞E3-1-1。 五倍子和诃子分别以1∶1、1∶2、1∶3、1∶4、1∶5和2∶1、3∶1、4∶1、5∶1的配比混合，测定其对鱼类病原菌联合抑菌效果。结果表明，在联合用药中，五倍子比例越高，抑菌活性越好；当五倍子比例高于诃子时，联合效应均表现为协同或累加作用。对气单胞菌进行联合用药时，五倍子和诃子以2∶1的配比的抑菌活性最好；对于弧菌，以3∶1和4∶1的配比的抑菌活性最好。 用五倍子和诃子进行了不同提取方法和不同提取溶剂的提取，测定了其提取率及提取物抑菌活性。结果表明，提取率均以超声波法和95％乙醇提取最高。超声波法提取率是溶剂回流法的1.1倍～2.4倍；用95％乙醇提取五倍子，提取率可达73.39％以上；诃子可达13％以上．从提取物抑菌活性来看，对于气单胞菌，各种溶剂提取五倍子，其抑菌活性差异不大，回流法和超声波法提取也差异不大。提取诃子时则以95％乙醇提取和超声波法较好。对于弧菌，不论是提取五倍子还是诃子，均以乙酸乙酯的提取物的抑菌活性最强，同时以超声波法最好。更多还原

【Abstract】 It is an effective way to search new antibacterial activity substance from Chinese herbs to control baeteriosis of fishes. There are abundant resources from herbs in our country, and it is easy to gain. In the reaserch, 101 sorts of common herbs are tested, and the bioassayed bacteria are Aeromonas hydrophila ST-3-3, Aeromonas caviae CSS-4-2, Vibrio anguillarum E3-J-1, Vibrio carchariae PHK and PUS. We deeply studied Gulla chinensis and Fructus Chebulae which have strong antimicrobial activity. The results as the following:We determined antibacterial activity of 101 sorts of herbs. It was found that 68 sorts of Chinese herbs have antibacterial action to at least one strain, and there are 48 Chinese herbs have strong antibacterial activity, such as Gulla chinensis, Fructus Chebulae, Bulbus Allii and so on. The widest antimicrobial spectrum is Gulla chinensis, Fructus Chebulae and Bulbus Allii. All tested bacteria are inhibited by them. The max bacteriostatic circle diameter of inhibiting to bacteria of Gulla chinensis is 32.84mm, and the MIC is 0.0488mg/ml; The max bacteriostatic circle diameter of inhibiting to bacteria of Fructus Chebulae is 28.72mm, and the MIC is 1.18725mg/ml; The max bacteriostatic circle diameter of inhibiting to bacteria of Bulbus Allii is 28.80mm, and the MIC is 0.78125mg/ml.Gulla chinensis and Fructus Chebulae or their vary proportion mixture have strong microbiostatic action to fish pathogenic bacteria. The MIC and MBC of Gulla chinensis is between 0.048828mg/ml and 0.78125mg/ml, and between 0.097656mg/ml and 0.78125mg/ml respectively. The MIC and MBC of Fructus Chebulae is between 0.097656mg/ml and 6.25mg/ml, and between 0.195313mg/ml and 25 mg/ml. The antibacterial activity of Gulla chinensis or Fructus Chebulae to 5 , strains is: PHK>PHS>CSS-4-2>ST-3-3>E3-l-l.The joint effect that Gulla chinensis incorporated with Fructus Chebulae to different bacteria indicated that mixture showed strong antimicrobial activity to all strains. The more percent of Gulla chinensis, the more antimicrobial activity will be.When the proportionment of Gulla chinensis is more than Fructus Chebulae, the joint effect exhibit synergy or additivism. To Aeromonas spp, the most effective antibacterial activity of mixture is 2 times Gulla chinensis with 1 time Fructus Chebulae. To Vibrio spp, the most effective antibacterial activity of mixture is 3 : 1 or 4: 1.The extract efficiency of supersonic method was superior to the other methods by the same solvent. Used same method, the extract efficiency of 95% ethanol is the best. The extract efficiency of supersonic method is 1.1~1.2 times to the efficiency of solvent circumfluence method. By 95% ethanol, the extract rate of Gulla chinensis is over 73.39%. The extract rate ofFructus Chebulae is over 13%. As far as antibacterial activity was concerned, the extracts of Gulla chinensis by every method was similar to Aeromonas spp. The result is also similar to the extracts by different solvent to Aeromonas spp. To Vibrio spp, the best method to extract Fructus Chebulae is by 95% ethanol and Supersonic method. Both the antibacterial activity of Gulla chinensis and Fructus Chebulae is best by ethyl acetate method and supersonic method.更多还原