LEPAE-Department of Chemical Engineering, Faculty of Engineering, University of Porto, Portugal.
J Appl Microbiol. 2012 Aug;113(2):373-82. doi: 10.1111/j.1365-2672.2012.05332.x. Epub 2012 May 31.
This work describes the effects of the presence of nonconjugative plasmids in Escherichia coli cells forming biofilms on a flow cell system under turbulent conditions.
The pET28 and pUC8 plasmids were separately used to transform E. coli JM109(DE3). Biofilm formation, removal and antimicrobial susceptibility to the cationic biocide benzyldimethyldodecylammonium chloride (BDMDAC) were assessed. Transformed cells formed thicker biofilms with higher cell densities, and the metabolic activity was higher whereas nontransformed cells had higher viabilities. Biocide treatment was not efficient for biofilm removal but was effective for cell killing. Biofilms formed by nontransformed cells were less affected by the treatment.
Cell transformation with the tested plasmids has significant impacts on biofilm formation, cell viability, metabolic activity and resistance to biocide treatment. Our results show that in biofilm studies involving deletion/complementation experiments, a control with the strain carrying a plasmid devoid of the gene under investigation must be included so that the real effects of the genetic manipulation are not biased by the presence of the plasmid backbone.
This is the first report where the presence of nonconjugative plasmids is assessed in flow conditions analysing biofilm formation, removal and antimicrobial susceptibility of high cell-density biofilms.
本工作描述了非接合质粒在形成生物膜的大肠杆菌细胞中的存在对湍流条件下流动池系统的影响。
分别使用 pET28 和 pUC8 质粒转化大肠杆菌 JM109(DE3)。评估了生物膜形成、去除以及对阳离子杀生剂苯扎氯铵(BDMDAC)的抗菌敏感性。转化细胞形成了更厚的生物膜,具有更高的细胞密度和更高的代谢活性,而非转化细胞具有更高的存活率。杀生物剂处理对生物膜去除无效,但对细胞杀伤有效。非转化细胞形成的生物膜受处理的影响较小。
用测试质粒进行细胞转化对生物膜形成、细胞活力、代谢活性和抗杀生物剂处理有显著影响。我们的结果表明,在涉及缺失/互补实验的生物膜研究中,必须包括携带无研究基因质粒的菌株作为对照,以避免质粒骨架的存在对遗传操作的真实影响产生偏差。
这是第一个在流动条件下评估非接合质粒存在对高细胞密度生物膜形成、去除和抗菌敏感性影响的报告。