Assessing the viability of bacterial species in drinking water by combined cellular and molecular analyses.

2.50
HDL Handle:
http://hdl.handle.net/10033/244591
Title:
Assessing the viability of bacterial species in drinking water by combined cellular and molecular analyses.
Authors:
Kahlisch, Leila; Henne, Karsten; Gröbe, Lothar; Brettar, Ingrid; Höfle, Manfred G
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 phylotypes 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.
Affiliation:
Department of Vaccinology and Applied Microbiology, Helmholtz Centre for Infection Research, Inhoffenstrasse 7, 38124 Braunschweig, Germany.
Citation:
Assessing the viability of bacterial species in drinking water by combined cellular and molecular analyses. 2012, 63 (2):383-97 Microb. Ecol.
Journal:
Microbial ecology
Issue Date:
Feb-2012
URI:
http://hdl.handle.net/10033/244591
DOI:
10.1007/s00248-011-9918-4
PubMed ID:
21845446
Type:
Article
Language:
en
ISSN:
1432-184X
Appears in Collections:
Publications of Dept. Vaccinology (VAC)

Full metadata record

DC FieldValueLanguage
dc.contributor.authorKahlisch, Leilaen_GB
dc.contributor.authorHenne, Karstenen_GB
dc.contributor.authorGröbe, Lotharen_GB
dc.contributor.authorBrettar, Ingriden_GB
dc.contributor.authorHöfle, Manfred Gen_GB
dc.date.accessioned2012-09-18T08:22:06Z-
dc.date.available2012-09-18T08:22:06Z-
dc.date.issued2012-02-
dc.identifier.citationAssessing the viability of bacterial species in drinking water by combined cellular and molecular analyses. 2012, 63 (2):383-97 Microb. Ecol.en_GB
dc.identifier.issn1432-184X-
dc.identifier.pmid21845446-
dc.identifier.doi10.1007/s00248-011-9918-4-
dc.identifier.urihttp://hdl.handle.net/10033/244591-
dc.description.abstractThe 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 phylotypes 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.en_GB
dc.language.isoenen
dc.rightsArchived with thanks to Microbial ecologyen_GB
dc.subject.meshBacteriaen_GB
dc.subject.meshColony Count, Microbialen_GB
dc.subject.meshDNA Fingerprintingen_GB
dc.subject.meshDNA, Bacterialen_GB
dc.subject.meshDrinking Wateren_GB
dc.subject.meshFlow Cytometryen_GB
dc.subject.meshMicrobial Viabilityen_GB
dc.subject.meshMolecular Sequence Dataen_GB
dc.subject.meshOrganic Chemicalsen_GB
dc.subject.meshPolymerase Chain Reactionen_GB
dc.subject.meshPolymorphism, Single-Stranded Conformationalen_GB
dc.subject.meshPropidiumen_GB
dc.subject.meshRNA, Bacterialen_GB
dc.subject.meshRNA, Ribosomal, 16Sen_GB
dc.titleAssessing the viability of bacterial species in drinking water by combined cellular and molecular analyses.en
dc.typeArticleen
dc.contributor.departmentDepartment of Vaccinology and Applied Microbiology, Helmholtz Centre for Infection Research, Inhoffenstrasse 7, 38124 Braunschweig, Germany.en_GB
dc.identifier.journalMicrobial ecologyen_GB
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