摘要
[目的]为了寻求无毒害且不产生耐药性的环保生物药物,取代当前抗生素的过量使用。[方法]从新鲜健康草鱼肠道中分离纯化一株芽孢杆菌,按常规细菌生化试验方法做生化鉴定。设计2对引物,进行16S r RNA检测与测序分析,按照琼脂扩散法做药敏试验。采用牛津杯法做对金黄色葡萄球菌体外抑制作用试验。[结果]分离菌与枯草芽孢杆菌生化特性基本一致,同源性为100%,确定为枯草芽孢杆菌;分离菌对阿米卡星、头孢氨苄、环丙沙星等敏感,对阿莫西林、氨苄西林、青霉素G等不敏感;对金黄色葡萄球菌抑菌效果明显,同时测出最小抑菌浓度(MIC)为2.8×108×2-5/ml,最小杀菌浓度(MBC)为2.8×108×2-2/ml。[结论]所分离的枯草芽孢杆菌可用于预防和治疗由金黄色葡萄球菌引起的疾病,减少抗生素的使用。
[Objective] To produce drug resistance, seek non-toxic environmental so as to change the current biological drugs that did not excessive use of antibiotics. [Methods] A strain of Bacillus was purified and isolated from fresh and healthy in- testines of grass carps. Biochemical identification was carried out by conventional bacterial biochemical test method. Two pairs of primers were designed, 16S rRNA detection and sequencing analysis were carried out. Drug sensitive test was carried out by agar diffusion method. In vitro inhibition test on Staphylococcus aureus was carried out by Oxford cup method. [Results] The isolated bacterium had basically the same biochemical characters as Bacillus subtilis; and the homology reached 100%. Thus, the isolated bacterium was identified to be Bacillus subtilis. It was insensitive to amoxicillin, ampicillin, penicillin G and so on, but sensitive to amikacin, cefalexin, ciprofloxacin and cefradine. The inhibitory effects of Bacillus subtilis on Staphylococ- cus aureus were significant. The minimum inhibitory concentration （MIC） was 2.8×10^8×2^-5/ml and minimum bactericidal concentration （MBC） was 2.8×10^8×2^-2/ml. [Conclusions] The isolated Bacillus subtilis could be used to prevent and control diseases caused by Staphylococcus aureus, and reduce the abuse of antibiotics.
基金
Supported by the Cooperation Subject(09003699)
the Project of Jiangxi Education Department(GJJ12237)
the Project of Science and Technology Department of Jiangxi(20122BBF60082)~~
