Colorimetric method for identifying plant essential oil components that affect biofilm formation and structure
Colorimetric method for identifying plant essential oil components that affect biofilm formation and structure
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DOI：
10.1128/aem.70.12.6951-6956.2004
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发表时间：
2004-12-01
影响因子：
4.4
通讯作者：
Gilbert, ES
中科院分区：
文献类型：
--
作者：
Niu, C;Gilbert, ES
特异性生物膜形成（SBF）测定，一种基于结晶紫染色的技术，被开发用于定位影响生物膜形成的植物精油及其组分。SBF分析确定，肉桂，肉桂，香茅油差异影响生长正常化的生物膜形成大肠杆菌。相应的精油的主要成分的SBF测定的检查显示，肉桂醛降低生物膜的形成相比，在Luria-Bertani肉汤中生长的生物膜，丁香酚没有导致的变化，香茅醇增加的SBF。为了评价这些结果，采用了两种基于显微镜的测定。首先，利用共聚焦激光扫描显微镜（CLSM）对E.用COMSTAT图像分析软件对流式细胞仪中培养的大肠杆菌生物膜进行定量分析。表征生物膜发展的五个参数的总体趋势证实了SBF测定的结果。其次，通过直接显微镜测量生长归一化粘附的测定结果与SBF测定和CLSM成像的结果一致。活性染色表明，与对嗜酸性细胞的毒性相比，精油组分对生物膜中细胞的毒性降低，但对E.肉桂醛暴露后的大肠杆菌。肉桂醛对E.杆菌三个假单胞菌属物种暴露于肉桂醛，丁香酚，香茅醇的SBF分析显示不同的反应。SBF测定可用作寻找影响生物膜形成和结构的植物精油及其组分的初始步骤。
The specific biofilm formation (SBF) assay, a technique based on crystal violet staining, was developed to locate plant essential oils and their components that affect biofilm formation. SBF analysis determined that cinnamon, cassia, and citronella oils differentially affected growth-normalized biofilm formation by Escherichia coli. Examination of the corresponding essential oil principal components by the SBF assay revealed that cinnamaldehyde decreased biofilm formation compared to biofilms grown in Luria-Bertani broth, eugenol did not result in a change, and citronellol increased the SBF. To evaluate these results, two microscopy-based assays were employed. First, confocal laser scanning microscopy (CLSM) was used to examine E. coli biofilms cultivated in flow cells, which were quantitatively analyzed by COMSTAT, an image analysis program. The overall trend for five parameters that characterize biofilm development corroborated the findings of the SBF assay. Second, the results of an assay measuring growth-normalized adhesion by direct microscopy concurred with the results of the SBF assay and CLSM imaging. Viability staining indicated that there was reduced toxicity of the essential oil components to cells in biofilms compared to the toxicity to planktonic cells but revealed morphological damage to E. coli after cinnamaldehyde exposure. Cinnamaldehyde also inhibited the swimming motility of E. coli. SBF analysis of three Pseudomonas species exposed to cinnamaldehyde, eugenol, or citronellol revealed diverse responses. The SBF assay could be useful as an initial step for finding plant essential oils and their components that affect biofilm formation and structure.