Abstract

The last decades have seen extensive scientific and technological improvements in many fields of microbiology and molecular biology. Correspondingly, flow cytometry—a rapid, precise and straightforward method for cultivation-independent detection of cells in liquids—has been a major topic in aquatic microbiology and drinking water analysis. Flow cytometry provides information at the single-cell level, including total cell counts, size measurements, nucleic acid content and bacterial viability and activity. While regulatory requirements for water testing rely on cultivation-based methods, flow cytometry can be considered a powerful tool to complement standard procedures.This article provides insights into the methodology and applicability of flow cytometry in the field of microbiological drinking water analysis and presents an overview on several case studies that cover a broad range of different objectives. The later are comprised of a study on flow cytometric characterization of Austrian drinking water resources, of an example for advanced data analysis methods of flow cytometric data, of a study on monitoring microbial regrowth within the distribution network, of an exemplary case of the application of online flow cytometry for high-frequency monitoring and of an introduction to the combination of flow cytometry and sequencing information.Finally, it is argued that due to the high microbiological variability of different water resources, unusual changes of flow cytometric parameters, rather than specific limits, could act as an indicator for further investigation. In this way flow cytometry can provide a good basis for risk assessments in water safety plans. The application of flow cytometry still remains utility-specific and a huge need for standardization of data analysis and interpretation exists in order to achieve a better cooperation of water utilities.

Highlights

  • Flow cytometry (FC) refers to the analysis of suspended particles based on scattered light and/or fluorescenceOriginalarbeit when passing a laser beam

  • Recent studies highlighted FC as a rapid, precise and straightforward method for detecting changes and characterizing the bacterial community in water samples (Hammes, Berney, and Egli 2010; Prest et al 2013; Gillespie et al 2014; Van Nevel et al 2017; Safford and Bischel 2019). It can provide a broad range of information at the single-cell level, including total cell counts, size measurements, nucleic acid content and aspects in bacterial viability and activity (Hammes, Berney, and Egli 2010; Prest et al 2013)

  • The microbiological analysis of water and drinking water mainly relies on culture-based parameters to ensure safe and biological stable water

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Summary

Introduction

Flow cytometry (FC) refers to the analysis of suspended particles based on scattered light and/or fluorescenceOriginalarbeit when passing a laser beam. Recent studies highlighted FC as a rapid, precise and straightforward method for detecting changes and characterizing the bacterial community in water samples (Hammes, Berney, and Egli 2010; Prest et al 2013; Gillespie et al 2014; Van Nevel et al 2017; Safford and Bischel 2019). It can provide a broad range of information at the single-cell level, including total cell counts, size measurements, nucleic acid content and aspects in bacterial viability and activity (Hammes, Berney, and Egli 2010; Prest et al 2013). While no regulatory limits for drinking water have been set so far, quantification of total bacterial numbers can give important additional information to the understanding of microbial survival and growth during drinking water treatment and distribution (Hammes et al 2008) and to monitoring source water quality in high temporal resolution (Besmer and Hammes 2016)

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