中华草龟荧光假单胞菌的分子鉴定及分析

Molecular Identification and Analysis of Pseudomonas Fluorescens in Chinemys reevesii

从患病草龟病灶分离到几株优势菌,对该菌进行革兰氏染色、镜检,在普通肉汤培养基上进行观察。对分离菌株进行生化鉴定及16S rRNA基因序列的克隆、测定,并对其DNA进行测序,在NCBI上进行比对,构建系统进化树,对该菌的基因序列进行分析比对,基于形态学、生理学、生物化学和基因序列分析,将这些菌株鉴定为荧光假单胞菌。药敏实验结果表明该菌对复方新诺明、氧氟沙星、恩诺沙星、诺氟沙星高度敏感,对卡那霉素、四环素、庆大霉素、新霉素、氟苯尼考、氯霉素中度敏感,对头孢噻吩、甲氧苄啶有耐药性。

Several strains of dominant bacteria were isolated from the diseased tissue of Chinemys reevesii. Gram staining and microscopic examination were performed on the bacteria, and observed on ordinary broth medium.From the isolated strains, biochemical identification and cloning and determination of 16S rRNA gene sequences have been done. The DNA was sequenced, aligned on NCBI, and a phylogenetic tree was constructed. The gene sequence of the strain was analyzed and compared. The morphology, physiological and biochemical characteristics and gene sequence analysis results of the strain were combined to confirm that these strains were Pseudomonas fluorescens. The results of drug susceptibility test showed that the strain was highly sensitive to compound sulfamethoxazole, ofloxacin, enrofloxacin and norfloxacin, and to kanamycin, tetracycline, gentamicin, neomycin,florfenicol, chloramphenicol moderately sensitive, and resistant to cefotaxime, trimethoprim.