Application of Bacterial Bioluminescence To Assess the Efficacy of Fast-Acting Biocides
Application of Bacterial Bioluminescence To Assess the Efficacy of Fast-Acting Biocides
复制标题
DOI：
10.1128/aac.00489-11
复制
发表时间：
2011-11-01
影响因子：
4.9
通讯作者：
Reynolds, Darren M.
中科院分区：
文献类型：
--
作者：
Robinson, Gareth M.;Tonks, Katherine M.;Reynolds, Darren M.
传统的微生物学技术用于提供一系列杀生物剂的杀灭率和杀灭程度的可靠数据。然而，这些技术提供的关于速效杀生物剂在非常短的时间域内的初始杀灭率的数据非常有限。本研究描述了一种表达发光杆菌lux操纵子的重组大肠杆菌菌株作为全细胞生物传感器的应用。光发射与细菌代谢直接相关;因此，通过监测光输出，可以评估速效杀菌剂的影响。电化学活化溶液（ECAS），漂白剂，Virkon，和乙醇进行了评估，在三个浓度（1%，10%，80%）的有机污染的存在。在2秒的时间过程中，80%ECAS产生的光输出的最大减少在没有有机负载，但强烈抑制其存在。在有机土壤中，80%的乙醇优于所有测试的杀生物剂。在测定的时间过程内，在匹配的浓度下，Bleach和Virkon产生类似的生物发光减少。它还表明，该测定可用于快速评估有机污染的影响。使用生物发光细菌作为全细胞生物报告者允许在应用的毫秒内评估快速作用的杀生物剂的相对功效。该测定可用于研究短时间或延长时间域的活性，以确认生物报告基因的完全代谢抑制。此外，该测定法可以通过允许研究传统微生物学所排除的时间域上的活性来进一步阐明其作用机制。
Traditional microbiological techniques are used to provide reliable data on the rate and extent of kill for a range of biocides. However, such techniques provide very limited data regarding the initial rate of kill of fast-acting biocides over very short time domains. This study describes the application of a recombinant strain of Escherichia coli expressing the Photorhabdus luminescens lux operon as a whole-cell biosensor. Light emission is linked directly to bacterial metabolism; therefore, by monitoring light output, the impact of fast-acting biocides can be assessed. Electrochemically activated solutions (ECASs), bleach, Virkon, and ethanol were assessed at three concentrations (1%, 10%, 80%) in the presence of organic soiling. Over a 2-s time course, 80% ECAS produced the greatest reduction in light output in the absence of organic load but was strongly inhibited by its presence. Eighty percent ethanol outperformed all tested biocides in the presence of organic soil. Bleach and Virkon produced similar reductions in bioluminescence at matched concentrations within the time course of the assay. It was also demonstrated that the assay can be used to rapidly assess the impact of organic soiling. The use of bioluminescent bacteria as whole-cell bioreporters allows assessment of the relative efficacies of fast-acting biocides within milliseconds of application. The assay can be used to investigate activity over short or extended time domains to confirm complete metabolic inhibition of the bioreporter. Moreover, the assay may enable further elucidation of their mechanism of action by allowing the investigation of activity over time domains precluded by traditional microbiology.