Flow cytometry for rapid analysis of bacteriostatic versus bactericidal effects in Legionella pneumophila disinfection.

IF 3.8 2区 化学 Q1 BIOCHEMICAL RESEARCH METHODS
Yiao Liang, Lena Heining, Martin Elsner, Michael Seidel
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Abstract

Biocides, encompassing both oxidizing agents such as chlorine and non-oxidizing agents like quaternary ammonium compounds, are extensively employed to minimize the risk of Legionella pneumophila (L. pneumophila) outbreaks in evaporative cooling systems. Currently, biocide effectiveness to combat L. pneumophila is determined by cultivation test methods, which have the disadvantage of long testing times and fail to detect a viable but non-culturable status of bacteria. Cultivation-independent bacterial counting methods would, therefore, be superior for future analysis of biocide effects. Herein, we employed total cell counting and intact cell counting by flow cytometry (FCM) as rapid, cultivation-independent, and untargeted methods for effect-based analysis of biocide effectiveness. We used total cell counting and intact cell counting to detect DNA damage and membrane integrity changes in response to oxidizing sodium hypochlorite (NaClO) and non-oxidizing benzalkonium chloride (BAC). Biocide effect analysis over time was conducted on a dedicated cartridge-based flow cytometer for bacteria analysis, rqmicro.COUNT. The effects of NaClO and BAC on cell membranes and DNA could be well determined and distinguished. Process water contaminated by L. pneumophila was treated with different concentrations of NaClO and BAC, followed by regrowth experiments to determine bacteriostatic versus bactericidal effects. The results showed that a considerable reduction in intact cell count was related to the bacteriostatic effects of the treatment and the success of the disinfection of L. pneumophila according to the standard cultivation method. A significant decrease in total cell count indicating DNA damage was associated with bactericidal effects. Hence, we could show the potential of total cell counting and intact cell counting by FCM to be used as a rapid and effect-based analysis method for minimizing both health-related risks and biocide usage in process water.

流式细胞术快速分析嗜肺军团菌消毒抑菌与杀菌效果。
杀菌剂,包括氧化剂如氯和非氧化剂如季铵化合物,被广泛用于减少蒸发冷却系统中嗜肺军团菌爆发的风险。目前,杀菌剂对嗜肺乳杆菌的有效性是通过培养试验方法来确定的,这种方法的缺点是测试时间长,不能检测出有活力但不可培养的细菌状态。因此,独立于培养的细菌计数方法在将来的杀菌剂效果分析中将是优越的。在此,我们采用流式细胞术(FCM)的细胞总数和完整细胞计数作为快速、不依赖培养、非靶向的方法来分析杀菌剂的有效性。我们使用细胞总数和完整细胞计数来检测氧化性次氯酸钠(NaClO)和非氧化性苯扎氯铵(BAC)对DNA损伤和膜完整性的影响。在专用的细菌分析用流式细胞仪rqmicro.COUNT上进行了一段时间内的杀菌剂效果分析。NaClO和BAC对细胞膜和DNA的影响可以很好地测定和区分。用不同浓度的NaClO和BAC处理被嗜肺乳杆菌污染的工艺水,然后进行再生实验,以确定抑菌与杀菌效果。结果表明,完整细胞数量的大量减少与处理的抑菌效果和按标准培养方法消毒嗜肺乳杆菌的成功有关。细胞总数的显著减少表明DNA损伤与杀菌作用有关。因此,我们可以证明FCM的总细胞计数和完整细胞计数的潜力,作为一种快速和基于效果的分析方法,可以最大限度地减少与健康相关的风险和工艺用水中的杀菌剂使用。
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来源期刊
CiteScore
8.00
自引率
4.70%
发文量
638
审稿时长
2.1 months
期刊介绍: Analytical and Bioanalytical Chemistry’s mission is the rapid publication of excellent and high-impact research articles on fundamental and applied topics of analytical and bioanalytical measurement science. Its scope is broad, and ranges from novel measurement platforms and their characterization to multidisciplinary approaches that effectively address important scientific problems. The Editors encourage submissions presenting innovative analytical research in concept, instrumentation, methods, and/or applications, including: mass spectrometry, spectroscopy, and electroanalysis; advanced separations; analytical strategies in “-omics” and imaging, bioanalysis, and sampling; miniaturized devices, medical diagnostics, sensors; analytical characterization of nano- and biomaterials; chemometrics and advanced data analysis.
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