Shao Jing, Lu KeQiao, Tian Ge, Cui YanYan, Yan YuanYuan, Wang TianMing, Zhang XinLong, Wang ChangZhong
Laboratory of Microbiology and Immunology, School of Chinese and Western Integrative Medicine, Anhui University of Chinese Medicine, Hefei 230038, China.
Laboratory of Biochemistry and Molecular Biology, School of Chinese and Western Integrative Medicine, Anhui University of Chinese Medicine, Hefei 230038, China.
J Microbiol Methods. 2015 Feb;109:41-8. doi: 10.1016/j.mimet.2014.12.006. Epub 2014 Dec 17.
The assembly of a man-made gravity-supported free-flow biofilm incubator (GS-FFBI) was described, which was composed of a gas cushion injector and four incubators. The GS-FFBI had the characteristics of (i) a bottom-up flow direction, and (ii) lab-scale biofilm preparation without the use of a multichannel pump. Two opportunistic fungal strains, namely Candida albicans and Candida glabrata, were employed to incubate C. albicans and dual C. albicans-C. glabrata biofilms on the surface of medical-grade polyvinyl chloride perfusion tube. In terms of the results from {2, 3-bis (2-methoxy-4-nitro-5-sulfo-phenyl)-2H-tetrazolium-5-carboxanilide} (XTT) assay, dry weight measurement, colony-forming unit counting, susceptibility test, and scanning electron microscopy, it was demonstrated that GS-FFBI could form both stable single and dual Candida biofilms with no significant variations among the four incubators or the three daily incubations within 21h, and could operate for at least 96h smoothly with no contamination of stock medium. The results also indicated, for the first time, that C. albicans and C. glabrata might be co-existent competitively and symbiotically in the dual biofilms with flowing media. GS-FFBI would be a useful device to study in vitro morphological and physiological features of microbial biofilms in the medical settings.
描述了一种人造重力支持的自由流动生物膜培养箱（GS-FFBI）的组装，它由一个气垫注射器和四个培养箱组成。GS-FFBI具有以下特点：（i）自下而上的流动方向，以及（ii）在不使用多通道泵的情况下进行实验室规模的生物膜制备。使用两种机会性真菌菌株，即白色念珠菌和光滑念珠菌，在医用级聚氯乙烯灌注管表面培养白色念珠菌生物膜以及白色念珠菌-光滑念珠菌混合生物膜。根据[2,3-双（2-甲氧基-4-硝基-5-磺基苯基）-2H-四唑-5-羧基苯胺]（XTT）测定、干重测量、菌落形成单位计数、药敏试验和扫描电子显微镜的结果表明，GS-FFBI能够形成稳定的单一和混合念珠菌生物膜，在四个培养箱之间或21小时内的三次每日培养之间没有显著差异，并且可以平稳运行至少96小时，储备培养基无污染。结果还首次表明，在有流动培养基的混合生物膜中，白色念珠菌和光滑念珠菌可能以竞争和共生的方式共存。GS-FFBI将是研究医疗环境中微生物生物膜体外形态和生理特征的有用设备。
