鲫鱼肝源细菌的分离纯化与鉴定
毕业论文关键词：鲫鱼；分离纯化；文隆气单胞菌；生理生化试验；16S rRNA.
Isolation and Identification of Aeromonas veronii from Carassius aumtus Liver
Abstract: I isolated and purified gram-negative bacteria strain from the diseased carp (Carassius aumtus) liver, temporarily numbered JG. After strain morphological, physiological and biochemical and 16S rRNA sequences were cloned and identified ways and so on. The results showed that: JG strains of Gram-negative short rod, rounded on TSA medium, uplift, smooth-edged cream small colonies. JG strain does not ferment inositol, dulcitol, esculin, xylitol, DNA enzyme, arabinose, rhamnose, xylose, sorbitol, citrate, raffinose, Adonis alcohol, lactose, hydrogen sulfide; urea, sucrose, OF, arginine hydrolase, lysine, glucose gas, phenylalanine, peptone water, VP test positive; of ornithine, MR, gluconate test negative, preliminarily identified as Villon JG strains of Aeromonas. Using PCR amplification of 16S rRNA fragments were sequenced after TA cloning total length is 1502bp, by 16S rRNA sequencing find JG strains of Aeromonas Villon ATCC 35624 homology of about 99.93%, and further determine JG strains dimension Long Aeromonas. This paper is revealing fish liver microbial species provides a theoretical basis for research carp disease control and public health significance.
Key words:Carassius aumtus ,Isolation and Purification, Aeromonas veronii ，Physiological-biochemical test,16S rRNA.
目录
前言 1
1 实验部分 1
1.1 实验材料 2
1.2 实验方法 2
1.2.1细菌的分离纯化、形态观察及理化性质 2
1.2.2基因组DNA的提取 2
1.2.3 16S rRNA片段的PCR扩增 2
1.2.4琼脂糖凝胶电泳 3
1.2.5TA克隆与测序 3
2 结果与分析 3
2.1 细菌形态及理化特征 3
2.2 16S rRNA序列及发育树分析 4
3 讨论 6
4 结论 7
致谢 7
参考文献 7
前言 鲫鱼（Carassius aumtus）是日常生活中常见的淡水鱼,这是鱼的一种。鲫鱼是杂食性鱼类，但植食性较多，由于各种各样的水生植物营养丰富，一些果实和水生植物的种子是鲫鱼的最爱，另外一些藻类、螺蛳和小虾也是鲫鱼喜爱食用的食物。鲫鱼群居生活，常生活在饵料充足的水域。鲫鱼分布大江南北，全国各地的水域常年均有生产，属于经济型可食用的鱼种，由于物美价廉，受到大多数人民的青睐。在我国黄河地区、长江流域的农业大区均有大面积养殖，其中有著名的洱海鲫鱼[1]。近年来，随着对鲫鱼的人工繁殖技术的不断发展，鲫鱼本身具有可观的经济价值，随着人工集约化养殖的不断扩展，随后而来的鲫鱼的病害问题也日趋严重，抑制鲫鱼养殖业发展的同时又给养殖者带来了巨大的经济损失。对于鲫鱼肝脏内细菌的研究大多数都是集中在鲫鱼的繁殖和培育问题上，极少深入地去分析引起鲫鱼发病的原因，当前从鲫鱼体内分离出耶尔森氏菌[4]可使动物和人患腹泻和胃肠炎，并且还能引起心血管、呼吸系统等疾病；希瓦氏菌[5]；迟钝爱德华氏菌[6]可引起牙鲆腹部膨胀、突眼、体表充血和肠管脱出等，另有文隆气单胞菌 [7]可引起鲫鱼出血和腹水。