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为了确定牛粪发酵沼液中疑似菌株的生物特性和种属关系。采用Hungate滚管技术、厌氧培养、革兰氏染色镜检、PCR扩增技术和抗生素耐受性鉴定等方法进行分析。实验结果表明,牛粪发酵沼液分离到了一株革兰氏阳性菌,能够利用CO_2,最适生长温度为37℃,单独或成对存在。菌落呈灰白色,光滑,表面凸起。PCR扩增出1 200 bp的16S rDNA片段,测序结果表明,该分离菌为产甲烷短杆菌。
In order to confirm the biological characteristics and the genus relationship of the suspected strains in the cow manure fermentation biogas slurry. Using Hungate roll tube technology, anaerobic culture, Gram staining microscopy, PCR amplification and identification of antibiotic resistance and other methods were analyzed. The experimental results showed that a Gram-positive bacterium was isolated from cow dung fermentation biogas slurry, and the optimal temperature for growth was 37 ℃, which could be used alone or in pairs. Colonies were gray, smooth, raised surface. PCR amplification of a 16S rDNA fragment of 1 200 bp, sequencing showed that the isolated bacteria Brevibacterium parvum.