【作者】 崔萍；

【导师】 刘训理；

【作者基本信息】 山东农业大学 ， 特种经济动物饲养， 2008， 硕士

【摘要】 利用平板稀释培养法对缫丝废水处理池中的细菌进行了分离,通过紫外分光光度法对其降解家蚕蛹油的能力进行了测定。采用细菌基因组重复序列PCR技术(rep-PCR),对家蚕蛹油降解细菌进行了聚类分析。采用菌落形态、生理生化特征和16S rDNA序列同源性分析对家蚕蛹油降解细菌进行了鉴定。选用Shannom-Wiener指数(H’)、丰富度指数(S)、Pielou均匀度指数(J)和Simpson优势度指数(D),对家蚕蛹油降解细菌的生物多样性进行了分析。主要结果如下。从缫丝废水处理池中共分离得到57个对家蚕蛹油具有降解能力的细菌分离物。降解能力超过50%的细菌分离物有12个,其中H73、H23、H1、H5和H43在72h内对家蚕蛹油的降解率分别达到76.36%、62.04%、60.58%、53.85%和51.20%。家蚕蛹油降解细菌rep-PCR基因指纹分析图显示,57个家蚕蛹油降解细菌被分为44个聚类群。细菌鉴定结果表明,44个聚类群中的39个聚类群属于芽孢杆菌属(Bacillus sp.),1个聚类群为微杆菌属(Microbacterium sp.),2个聚类群为肠杆菌属(Enterobacter sp.),另外2个聚类群尚未获得其16S rDNA序列,有待进一步鉴定。各菌株的16S rDNA序列已在GenBank上注册。其中,芽孢杆菌属的细菌数量为1.4473×106CFU/mL,大大高于肠杆菌属和微杆菌属的细菌数量,为家蚕蛹油降解细菌中的优势菌群。多样性分析结果表明,芽孢杆菌属的多样性指数、丰富度指数、均匀度指数和优势度指数都高于其他两个属的细菌。更多还原

【Abstract】 The bacteria were isolated by the plate cultivation method from rolling wastewater and the degrading capability of those were studied with UV spectrophotometry. The repetitive-element PCR genomic fingerprinting (rep-PCR) were used to fingerprint pure DNA extracted from the isolates which can degrade silkworm pupal oil.Then they were identified by their morphological properties, physiological and biochemical characteristics and sequence analysis of 16S rDNA. The diversity of silkworm pupal oil degrading bacteria was analyzed by the Shannom-Wiener index, richness index, evenness index and domination index. The main results were as follows.57 isolates which can degrade the silkworm pupal oil were isolated from rolling wastewater. The number of isolates with the degrading ratio above 50% is 12. The strains H73, H23, H1, H5 and H43 showed higher degrading capability than others,which can degrade 76.36%,62.04%,60.58%,53.85% and 51.20% of total oil in 72 hours,respectively.Based on the rep-PCR fingerprints cluster analysis ,the degrading bacteria were divided into 44 clusters at the level of species.The identification results based on the results of phenotypic characteristics and phylogenetic analysis of 16S rDNA sequence showed that the 39 clusters of degrading-bacteria belonged to Bacillus sp.,one of them belonged to Microbacterium sp.,and two of them belonged to Enterobacter sp.. Besides,there are 2 clusters which need further identification. Their 16S rDNA sequences had been registered at Genbank.The number of bacteria belonged to Bacillus genera was 1.4473×106CFU/mL, which was larger than the other two generas. The diversity analysis showed that the bacteria belonged to Bacillus genera had higher Shannom-Wiener index, richness index, evenness index and domination index.更多还原