牛支原体生物被膜形成优势菌株的筛选及培养条件的优化  

Dominant Strain Screening of Mycoplasma bovis in Biofilm Formation and Optimization of Cultural Conditions

作　　者：高翔[1] 邢小勇[1] 伏小平[1] 温峰琴[1] 薛慧文[1] 魏彦明[1] 包世俊[1] GAO Xiang;XING Xiao-Yong;FU Xiao-Ping;WEN Feng-Qin;XUE Hui-Wen;WEI Yan-Ming;BAO Shi-Jun(College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,Chin)

机构地区：[1]甘肃农业大学动物医学院,兰州730070

出　　处：《农业生物技术学报》2018年第8期1449-1456,共8页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(No.31360620);甘肃省自然科学基金(No.1308RJZA235);国家肉牛/牦牛产业体系项目(No.CARS-37)

摘　　要：牛支原体(Mycoplasma bovis,Mb)是牛(Bos taurus)的重要致病性支原体,其生物被膜的形成在宿主细胞中产生持续性感染,使养牛业造成重大的经济损失。因此,本研究旨在通过对Mb不同菌株生物被膜形成能力的鉴定,筛选出1株生物被膜形成优势菌株,并对其培养条件进行优化,以期为Mb生物被膜相关的研究提供生物材料。首先应用微孔板定量检测法鉴定Mb PG45株、湖北株(HB株)、武威株(WW株)、定西株(DX株)和临洮株(LT株)生物被膜的形成能力;进而以生物被膜形成能力强的菌株作为模式菌株,完成对Mb生物被膜形成培养条件的优化。结果显示,Mb不同菌株形成生物被膜的能力不同,其中临洮株生物被膜形成能力最强,湖北株生物被膜形成能力次之,PG45株、武威株和定西株生物被膜形成能力最弱。取浓度为1.7E+08 CFU/mL的Mb液体培养物,按1∶10转接种于含0.01 mg/mL DNA、20%马血清和1%葡萄糖的Eaton培养基(pH 8.5),于37℃、5%CO2条件下培养72 h,Mb生物被膜的形成最佳。本研究筛选获得1株生物被膜形成能力强的Mb,并通过培养条件的优化,使之形成稳定且明显的生物被膜,从而为Mb生物被膜相关研究提供了良好的生物材料。Mycoplasma bovis （Mb） is an important pathogenic mycoplasma of cattle （Bos taurus）. The biofilm formation of Mb can cause persistent infection in host cells and cause great economic loss to cattle industry. Therefore, the purpose of this study is to screen a dominant strain of Mb biofilm formation by identifying the biofilm formation ability of different Mb strains, and then optimize the cultural conditions on biofilm formation of Mb. First of all, the ability of biofilm formation of Mb PG45 strain, Hubei （HB） strain, Wuwei （WW） strain, Dingxi （DX） strain, and Lintao （LT） strain were identified by polystyrene microtiter plate assay, and then the strain with strong biofilm forming ability was used as the model strain to optimize the culture conditions of Mb biofilm formation. The results showed that the ability of biofilm formation was different among different strains of Mb, among which the biofilm formation ability of LT strain was the strongest, the next was HB strain. And the biofilm formation ability of PG45 strain, WW strain and DX strain was the weakest. The optimal cultural conditions for biofilm formation of Mb was that the Mb cultures （1.7E＋08 CFU/mL） were inoculated at a dilution of 1∶10 into the Eaton medium containing 0.01 mg/mL fish sperm DNA, 20% serum of horse, 1% glucose and final pH 8.5, and then incubated at 37 ℃, with 5% CO2 for 72 h. Under the optimal cultural conditions, the biofilm formation ability of LT strain was sound. In this study, a dominant strain of Mb biofilm formation was obtained, and a stable and obvious biofilm was formed by optimizing the cultural conditions, which will provide excellent biomaterials for biofilm related study of Mb.

关 键 词：牛支原体(Mb) 生物被膜 优势菌株 影响因素 优化 

分 类 号：S852.62[农业科学—基础兽医学]