Department of Oto-Rhino-Laryngology, Head & Neck Surgery, Leiden University Medical Center, Leiden, The Netherlands.
Biofouling. 2010 Jul;26(5):519-26. doi: 10.1080/08927014.2010.489238.
The aim of this study was to demonstrate the presence of yeast and bacterial biofilms on the surface of tracheoesophageal voice prostheses (TVPs) by a double-staining technique with confocal laser scanning microscopy (CLSM). Biofilms of 12 removed TVPs were visualized by scanning electron microscopy, then stained with ConA-FITC and propidium iodide for CLSM. Microbial identification was by partial 16S rRNA gene analysis and ITS-2 sequence analysis. Microbial biofilms on the TVPs consisted of bacteria and filamentous cells. Bacterial cells were attached to the filamentous and unicellular yeast cells, thus forming a network. Sequence analyses of six voice prostheses identified the presence of a variety of bacterial and yeast species. In vivo studies showed that Klebsiella oxytoca and Micrococcus luteus efficiently attached to Candida albicans. CLSM with double fluorescence staining can be used to demonstrate biofilm formations composed of a mixture of yeast and bacterial cells on the surface of TVPs.
本研究旨在通过共聚焦激光扫描显微镜（CLSM）的双重染色技术证明气管食管语音假体（TVP）表面存在酵母和细菌生物膜。通过扫描电子显微镜观察了 12 个已去除的 TVP 的生物膜，然后用 ConA-FITC 和碘化丙啶进行 CLSM 染色。微生物鉴定通过部分 16S rRNA 基因分析和 ITS-2 序列分析进行。TVP 上的微生物生物膜由细菌和丝状细胞组成。细菌细胞附着在丝状和单细胞酵母细胞上，从而形成网络。对六个语音假体的序列分析确定存在多种细菌和酵母物种。体内研究表明，产酸克雷伯菌和藤黄微球菌能够有效地附着在白色念珠菌上。CLSM 双重荧光染色可用于证明 TVP 表面由酵母和细菌细胞混合组成的生物膜形成。
