一株工业腐败物中魏氏柠檬酸杆菌的分离鉴定及产生物膜特性分析

Identification and biofilm formation characterization of Citrobacter werkmanii isolated from industrial spoilage

摘要:

【目的】分离和鉴定工业腐败物中高产细菌生物膜菌株，并明确该菌的部分产膜特性。【方法】通过微孔板结晶紫染色法对分离的菌株进行产膜能力评价，根据菌落形态、生理生化特性和16S rRNA序列的系统进化树分析进行菌株鉴定；同时利用扫描电子显微镜(SEM)和结晶紫染色法分别研究材料及温度对该菌产膜特性和能力的影响。【结果】筛选出一株高产细菌生物膜菌株，经鉴定该菌为魏氏柠檬酸杆菌；其在玻璃、不锈钢和聚氯乙烯(PVC)材料表面均能形成生物膜；温度条件显著影响产膜能力，在30 °C时，菌株在PVC材料表面形成生物膜能力最强。【结论】工业腐败物中含有高产细菌生物膜菌株，并且产膜受附着物和温度影响。

关键词:魏氏柠檬酸杆菌，鉴定，扫描电镜，生物膜，温度

Abstract:

[Objective] A microorganism from the samples of industrial spoilage with high-yield bacterium biofilm was isolated and identified. Meanwhile studies on characterization of biofilm formation were also carried out. [Methods] First of all, crystal violet stain was used to evaluate the ability of the biofilm-forming of screened strains in 96-cell microplates. And then, colonial morphology, physio-biochemical tests and 16S rRNA sequence analysis were adopted to determine the phylogenetic position of the isolated strains. Finally, the effects of attachment materials and culture temperature on the characterization of biofilm formation were also researched using scanning electron microscope (SEM) and crystal violet stain respectively. [Results] The best biofilm-forming strain was identified and clarified into Citrobacter werkmanii. And biofilm could be formed on the surfaces of glass, stainless steel and polyvinylchloride (PVC) respectively. Moreover the characterization of biofilm could be significantly influenced by temperature and the best biofilm was found on the PVC at 30 °C no matter which materials. [Conclusion] In industrial spoilage, the biofilm-forming strains could be found and its ability to produce biofilm could also be affected by materials and temperature.

Key words:Citrobacter werkmanii, Identification, SEM, Biofilm, Temperature