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Assessing the Viability of Bacterial Species in Drinking Water by Combined Cellular and Molecular Analyses

  • Microbiology of Aquatic Systems
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Abstract

The question which bacterial species are present in water and if they are viable is essential for drinking water safety but also of general relevance in aquatic ecology. To approach this question we combined propidium iodide/SYTO9 staining (“live/dead staining” indicating membrane integrity), fluorescence-activated cell sorting (FACS) and community fingerprinting for the analysis of a set of tap water samples. Live/dead staining revealed that about half of the bacteria in the tap water had intact membranes. Molecular analysis using 16S rRNA and 16S rRNA gene-based single-strand conformation polymorphism (SSCP) fingerprints and sequencing of drinking water bacteria before and after FACS sorting revealed: (1) the DNA- and RNA-based overall community structure differed substantially, (2) the community retrieved from RNA and DNA reflected different bacterial species, classified as 53 phylotypes (with only two common phylotypes), (3) the percentage of phylotpes with intact membranes or damaged cells were comparable for RNA- and DNA-based analyses, and (4) the retrieved species were primarily of aquatic origin. The pronounced difference between phylotypes obtained from DNA extracts (dominated by Betaproteobacteria, Bacteroidetes, and Actinobacteria) and from RNA extracts (dominated by Alpha-, Beta-, Gammaproteobacteria, Bacteroidetes, and Cyanobacteria) demonstrate the relevance of concomitant RNA and DNA analyses for drinking water studies. Unexpected was that a comparable fraction (about 21%) of phylotypes with membrane-injured cells was observed for DNA- and RNA-based analyses, contradicting the current understanding that RNA-based analyses represent the actively growing fraction of the bacterial community. Overall, we think that this combined approach provides an interesting tool for a concomitant phylogenetic and viability analysis of bacterial species of drinking water.

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Acknowledgments

This work was supported by funds from the European Commission for the HEALTHY WATER project (FOOD-CT-2006-036306). The authors are solely responsible for the content of this publication. It does not represent the opinion of the European Commission. The European Commission is not responsible for any use that might be made of data appearing therein. We thank Josefin Draheim and Julia Strömpl for outstanding technical support.

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Authors and Affiliations

  1. Department of Vaccinology and Applied Microbiology, Helmholtz Centre for Infection Research (HZI), Inhoffenstrasse 7, 38124, Braunschweig, Germany

    Leila Kahlisch, Karsten Henne, Ingrid Brettar & Manfred G. Höfle

  2. Department of Cell Biology, Helmholtz Centre for Infection Research (HZI), Inhoffenstrasse 7, 38124, Braunschweig, Germany

    Lothar Gröbe

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  1. Leila Kahlisch
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  2. Karsten Henne
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  3. Lothar Gröbe
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  4. Ingrid Brettar
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  5. Manfred G. Höfle
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Correspondence to Ingrid Brettar.

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Kahlisch, L., Henne, K., Gröbe, L. et al. Assessing the Viability of Bacterial Species in Drinking Water by Combined Cellular and Molecular Analyses. Microb Ecol 63, 383–397 (2012). https://doi.org/10.1007/s00248-011-9918-4

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  • DOI: https://doi.org/10.1007/s00248-011-9918-4

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