【作者】 郑艳红；

【导师】 谢新民； 肖调义；

【作者基本信息】 湖南农业大学 ， 水产养殖， 2008， 硕士

【摘要】 三角帆蚌（Hyriopsis cumingii）是我国淡水育珠的主要蚌种,也是我国特有的河蚌资源,广泛分布在我国东部、南部及中部地区,尤以我省洞庭湖以及中型湖泊分布较多。本研究通过从湖南省常德市的几个疫区获得的病蚌和健康蚌进行研究,旨在找出病原,筛选出对于防治此类细菌性疾病的有效药物,培育优良的三角帆蚌种质资源,从而提高淡水珍珠的产量。1.病原菌的分离、鉴定及确定致病菌。从患病三角帆蚌的不同组织取欲检病料,对病原微生物进行分离、纯化及生化鉴定,得到温和气单胞菌、产碱假单胞菌、粘质沙雷菌、产酸克雷伯菌、解鸟氨酸克雷伯菌、洋葱伯克霍尔德氏菌,并通过攻毒试验确定温和气单胞菌和产碱假单胞菌为致病菌。2.中草药和抗生素的筛选。采用纸片扩散法测定抑菌直径,微量稀释法测定最低抑菌浓度（MIC）和最低杀菌浓度（MBC）,通过对30种中草药和18种抗生素的抑菌活性的测定,得出:在1g/ml的浓度下,黄连对温和气单胞菌和产碱假单胞菌的抑菌直径分别达到20.13mm、16.29mm,MIC分别为0.0125g/ml、0.0104g/ml,MBC分别为0.0125g/ml、0.0208g/ml;乌梅的抑菌直径分别达到18.26mm、20.38mm,MIC分别为0.0125g/m、0.0104g/ml,MBC分别为0.0208g/ml、0.0167g/ml。抗生素中以洛美沙星和左氧氟沙星活性最强,洛美沙星对温和气单胞菌和产碱假单胞菌的抑菌直径分别为25.64mm、25.24mm,MIC分别为1.667μg/ml、1.667μg/ml,MBC分别为2.083μg/ml、2.083μg/ml,左氧氟沙星的抑菌直径分别为25.48mm、24.67mm,MIC分别为1.667μg/ml、1.250μg/ml,MBC分别为2.917μg/ml、2.500μg/ml。3.采用正交试验筛选最优组方。结合前面筛选出的乌梅、黄连、洛美沙星和左氧氟沙星,按L₉（3⁴）设计正交试验,得出由0.5MIC黄连、1 MIC乌梅、1MIC洛美沙星、1MIC左氧氟沙星组成的药物复方,抑制温和气单胞菌的效果最好,洛美沙星和左氧氟沙星是影响抑菌圈直径的关键因子;而抑制产碱假单胞菌效果最好的组方为0.5MIC黄连、1 MIC乌梅、1MIC洛美沙星及1MIC左氧氟沙星,洛美沙星和左氧氟沙星是影响抑菌圈直径的关键因子。更多还原

【Abstract】 Hyriopsis cumingii is one of the main freshwater pear meussel scattering kinds of our country raised in fresh water.It is widely covered the Eastern,the Southern parts and central section.In this study,gaining the healthy and plague diseased mussels of Hriopsis cumingii lea from Changde city of Hunan province to find the resource of disease,and select the useful drug to control and prevent epidemic diseases of Ityriopsis cumingii.Clutivate excellent germplasm resources,so as to improve output of freshwater pear.The separation and identification of bacteria of Hyriopsis cumingii and determined pathogen:obtained diseased material from different tissues of diseased Hyriopsis cumingii.Separated、purified and identificated the pathogenic microorganism,we got 6 kinds of bacteria,seudomonas alcaligenes,Aeromonas sobria,Serratia marcescens,etc.and determined Aeromonas sobria and seudomonas alcaligenes as pathogen that caused Hyriopsis cumingii disease.The selection of Chinses herbs and antibiotics:By way of paper diffuseion to determine the antibacterial diameter and microdilution method to determine the Minimum Inhibitory Concentration（MIC） and the Minimum Bactedacidal Concentration（MBC）.Studied the antibacterial activity of 30 kinds of Chinses herbs and 20 kinds of antibiotics.the result was,at the concentration of 1g/ml,the antibacterial diameter of Rhicoma Coptidis on Aeromonas sobria and seudomonas alcaligenes are 20.13mm、16.29mm,respectively,the MIC were 0.0125g/ml and 0.0104 g/ml,respectively,the MBC were 0.0125g/ml and 0.0208g/ml,respectively; the corresponding results of Fructus Mume were 18.26mm and 20.38mm, respectively,the MIC were 0.0125g/m and 0.0104g/ml,respectively,the MBC were 0.0208g/m and 0.0167g/ml,respectively.Ciprofloxacin and Levofloxacin had the higher antibacterial activity in the 18 kinds of antibiotics,the antibacterial diameter of Ciprofloxacin on Aeromonas sobria and seudomonas alcaligenes were 25.64mm and 25.24mm,respectively,the MIC were 1.667μg/ml、1.667μg/ml,respectively,the MBC were 2.083μg/ml、2.083μg/m,respectively,the corresponding results of Levofloxacinare25.48mm and 24.67mm,respectively,the MICare 1.667μg/ml andl.250μg/ml,respectively,the MBC were 2.917μg/ml、2.500μg/ml,respectively.By way of orthogonal experiment to determined combination:combinate Lomef loxacin,Levofloxacin,Rhicoma Coptidis and Fructus Mume that selected before, according to L₉（3⁴） to design orthogonal experiment,we descovered that the best combination to Aeromonas sobria were 0.5 MIC（0.005g/ml）of Rhicoma Coptidis,1MIC（0.01g/ml） of Fructus Mume,1MIC（1.667μg/ml） of Iprofloxacin And 1MIC（1.667μg/ml） Levofloxacin,the best combination to seudomonas alcaligenes were 0.5 MIC（0.005g/ml） of Rhicoma Coptidis,1MIC（0.01g/ml） of Fructus Mume, 1MIC（1.25μg/ml） of Ievofloxacin And 1MIC（1.0μg/ml） Iprofloxacin.Iprofloxacin and levofloxacin were key factors that effected the antibacterial activity.更多还原