Laboratory of Microbial Ecology and Technology (LabMET), Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, 9000 Ghent, Belgium.
Appl Microbiol Biotechnol. 2010 Jun;87(1):331-41. doi: 10.1007/s00253-010-2526-2. Epub 2010 Mar 30.
The present study reports the disinfection effects of chemically and electrochemically dosed chlorine on two models for typical water-borne bacteria (Escherichia coli and Legionella beliardensis) by plating and flow cytometry (FCM) in combination with different fluorescence dyes. The residual effect on various cell functions, including cultivability, esterase activity, membrane polarization, and integrity, was tested at different free chlorine concentrations. In comparison, chemical disinfection yielded on average 60% more E. coli cells entering the viable but nonculturable (VBNC) state than electrochemical disinfection. Here, VBNC is defined as those cells with intact cell membrane but which cannot be cultured on solid nutrient agar plates. L. beliardensis was about five times more resistant to chlorine disinfection than E. coli. The results also suggested the two methods result in different disinfection mechanisms on L. beliardensis, i.e., chemically dosed chlorine targeted cell membrane integrity before enzyme activity, while electrochemically dosed chlorine acted the other way round. In addition, both bacteria lost the integrity of their cell membranes at three times lower chlorine concentration over a longer contact time (i.e., 40 vs. 10 min) by the chemical method. Our results showed that FCM is an appropriate tool to evaluate the effects of water disinfection and the percentage of cells in VBNC in a matter of hours. Electrochemical disinfection is suggested to be a favorable alternative for chemical disinfection.
本研究报告了化学和电化学剂量的氯对两种典型水源细菌(大肠杆菌和贝勒氏军团菌)的消毒效果,通过平板和流式细胞术(FCM)结合不同荧光染料进行检测。在不同游离氯浓度下,测试了对各种细胞功能(包括可培养性、酯酶活性、膜极化和完整性)的残留效果。相比之下,化学消毒使大肠杆菌进入活但不可培养(VBNC)状态的细胞数量平均比电化学消毒多 60%。在这里,VBNC 定义为那些细胞膜完整但不能在固体营养琼脂平板上培养的细胞。贝勒氏军团菌对氯消毒的抵抗力比大肠杆菌高约五倍。结果还表明,这两种方法对贝勒氏军团菌的消毒机制不同,即化学剂量的氯在酶活性之前靶向细胞膜完整性,而电化学剂量的氯则相反。此外,通过化学方法,两种细菌在接触时间更长(即 40 分钟对 10 分钟)、氯浓度低三倍的情况下,其细胞膜完整性丧失。我们的结果表明,FCM 是评估水消毒效果和 VBNC 状态下细胞百分比的合适工具,可在数小时内完成。电化学消毒被认为是化学消毒的一种有利替代方法。