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    摘要:鱼塘的健康养殖受到多种因素的制约,如鱼的种类以及鱼塘的面积大小等,同时,鱼塘水体中的细菌对鱼类存活更是带来重大影响。本文对徐州北郊鲫鱼鱼塘水体中细菌的多样性及致病菌对该鱼塘中鲫鱼存活状况的影响进行了研究,进一步实现鱼塘的科学管理。研究主要是通过对鲫鱼鱼塘水体中细菌的总DNA进行提取,经过PCR扩增并依据16S rRNA基因序列构建系统发育树,进行分子发育学研究,最后获得鱼塘水体中细菌物种的组成。它们分别为崛越氏芽孢杆菌(Bacillus horikoshii)、Aeromonas punctata、Bacillus thaonhiensis、溶血葡萄球菌(Staphylococcus haemolyticus)、弗氏志贺菌(Shigella flexneri)、印度芽孢杆菌(Bacillus indicus)、Fictibacillus phosphorivorans、Enterobbacter soli、Bacillus aryabhattai。经检测其中Aeromonas punctata、弗氏志贺菌(Shigella flexneri)及溶血性葡萄球菌(Staphylococcus haemolyticus)为致病菌。研究徐州北郊鱼塘水体中细菌多样性及其中致病菌对鱼类存活状况的影响不仅可以补充完善我国的鱼塘养殖环境中微生物物种资源,还为鱼塘健康养殖提供重要的参考依据。34355
    毕业论文关键词:多样性;阳性菌;阴性菌;致病菌;16S rRNA;系统发育树   
    Study on bacterial persity in fish pond water in the north of Xuzhou
    Abstract:Healthy breeding of fish ponds restricted by many factors, such as the types of fish and fish ponds area size, etc. At the same time, the bacteria in fish pond water for fish survival but also bring significant impact .This paper studies the influences of different bacteria to different fishes in the fish pond in the north of Xuzhou. The research is mainly through 16S rRNA gene sequencing of the fish pond water bacteria in the north of Xuzhou, and the 16S rRNA gene sequence analysis of phylogenetic tree construction, finally get the composition of bacterial species in water. After testing , We found that there were nine kinds of bacteria in fish pond water, they were Bacillus horikoshii、Aeromonas punctata、Bacillus thaonhiensis、Staphylococcus haemolyticus、Shigella flexneri、Bacillus indicus、Fictibacillus phosphorivorans、Enterobbacter soli、Bacillus aryabhattai. The Aeromonas punctata、Staphylococcus haemolyticus and Shigella flexneri were pathogenic bacteria.Study on the effect of bacterial persity and pathogen on fish survival status of the fish pond water in the north of Xuzhou can not only improve our microbial species resources of aquiculture environment, but also provide an important reference for the healthy breeding of the fish ponds.
    Key words: persity; Gram-negative bacteria; Gram-positive bacteria; pathogenic bacteria; 16S rRNA; Phylogenetic tree
    目录
    引言    1
    1材料与方法    2
    1.1    材料    2
    1.1.1    水样采集    2
    1.1.2    配制培养基    2
    1.1.3    主要仪器与试剂    2
    1.2  方法    2
    1.2.1    对菌种进行划线分离与纯化并斜面保存    2
    1.2.2    Gram stain染色    2
    1.2.3    细菌总DNA抽提及电泳分析    2
    1.2.4    细菌的PCR扩增    2
    1.2.5    PCR扩增产物电泳分析    3
    1.2.6    扩增产物胶回收    3
    1.2.7    测序    3
    1.2.8    构建系统发育树    3
    2结果与分析    3
    2.1    革兰氏染色结果    3
    2.2    系统发育树    3
    3讨论    5
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