Modes of cytometric bacterial DNA pattern: a tool for pursuing growth

Müller, Susann

doi:10.1111/j.1365-2184.2007.00465.x

Cited by 58 publications

(50 citation statements)

References 117 publications

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“…It appears justified to conclude that an increase in bacterial cell number within every subcommunity is caused by cell proliferation and upcoming new cell types (19). Such an approach based on DNA pattern analysis combined with the detection of TC-labeled polyphosphates can be regarded as highly reliable and quantitative, since there is no risk of active dye excretion by fixed cells (26,43).…”

Section: Discussionmentioning

confidence: 99%

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Dynamics of Polyphosphate-Accumulating Bacteria in Wastewater Treatment Plant Microbial Communities Detected via DAPI (4′,6′-Diamidino-2-Phenylindole) and Tetracycline Labeling

Günther

Trutnau

Kleinsteuber

et al. 2009

Appl Environ Microbiol

107

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Wastewater treatment plants with enhanced biological phosphorus removal represent a state-of-the-art technology. Nevertheless, the process of phosphate removal is prone to occasional failure. One reason is the lack of knowledge about the structure and function of the bacterial communities involved. Most of the bacteria are still not cultivable, and their functions during the wastewater treatment process are therefore unknown or subject of speculation. Here, flow cytometry was used to identify bacteria capable of polyphosphate accumulation within highly diverse communities. A novel fluorescent staining technique for the quantitative detection of polyphosphate granules on the cellular level was developed. It uses the bright green fluorescence of the antibiotic tetracycline when it complexes the divalent cations acting as a countercharge in polyphosphate granules. The dynamics of cellular DNA contents and cell sizes as growth indicators were determined in parallel to detect the most active polyphosphate-accumulating individuals/subcommunities and to determine their phylogenetic affiliation upon cell sorting. Phylotypes known as polyphosphate-accumulating organisms, such as a "Candidatus Accumulibacter"-like phylotype, were found, as well as members of the genera Pseudomonas and Tetrasphaera. The new method allows fast and convenient monitoring of the growth and polyphosphate accumulation dynamics of not-yet-cultivated bacteria in wastewater bacterial communities.

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Section: Discussionmentioning

confidence: 99%

“…Staining of the bacterial DNA gives information on bacterial growth rates (26,28) and therefore on activity states of PAOs when analyzed in combination with TC. DAPI staining alone of both DNA and polyphosphate granules did not result in a reliable analysis of the two characteristics.…”

Section: Discussionmentioning

confidence: 99%

Dynamics of Polyphosphate-Accumulating Bacteria in Wastewater Treatment Plant Microbial Communities Detected via DAPI (4′,6′-Diamidino-2-Phenylindole) and Tetracycline Labeling

Günther

Trutnau

Kleinsteuber

et al. 2009

Appl Environ Microbiol

107

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“…growing cells exhibiting more than one genome in average, whereas stationary cells contain only the minimum number of genomes because of starvation (36).…”

Section: Original Articlementioning

confidence: 99%

A flow cytometric method for viability assessment of Staphylococcus aureus and Burkholderia cepacia in mixed culture

et al. 2012

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Mixed bacterial communities are commonly encountered in microbial infections of humans. Knowledge on the composition of species and viability of each species in these communities allows for a detailed description of the complexity of interspecies dynamics and contributes to the assessment of the severity of infections. Several assays exist for quantification of specific species in mixed communities, including analysis of quantitative terminal restriction fragment length polymorphisms. While this method allows for species-specific cell enumeration, it cannot provide viability data. In this study, flow cytometry was applied to assess the viability of Staphylococcus aureus and Burkholderia cepacia in mixed culture by membrane integrity analysis using SYBR® Green I and propidium iodide staining. Both bacteria are relevant to pulmonary infections of cystic fibrosis patients. Fluorescence staining was optimized separately for each species in pure culture due to differences between species in cell wall structure and metabolic capabilities. To determine viability of species in mixed culture, a protocol was established as a compromise between optimum conditions determined before for pure cultures. This protocol allowed the detection of viable and dead cells of both species, exhibiting an intact and a permeabilized membrane, respectively. To discriminate between S. aureus and B. cepacia, the protocol was combined with Gram-specific fluorescent staining using wheat germ agglutinin. The established three-color staining method was successfully tested for viability determination of S. aureus and B. cepacia in mixed culture cultivations. In addition, growth of both species was monitored by quantitative terminal restriction fragment length polymorphisms. The obtained data revealed alterations in viability during cultivations for different growth phases and suggest interspecies effects in mixed culture. Overall, this method allows for rapid simultaneous Gram-differentiation and viability assessment of bacterial mixed cultures and is therefore suitable for the analysis of dynamics of mixed communities of medical, environmental, and biotechnological relevance.

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“…These clusters may represent cells of identical phylogenetic affiliation but also taxonomically different community members with identical or similar optical features. different cellular DNA contents and morphologies of the respective phylotype [44]. Typical distributions are shown in Fig.…”

Section: Community Structure By Cytometric Fingerprintingmentioning

confidence: 99%

High resolution single cell analytics to follow microbial community dynamics in anaerobic ecosystems

Müller

Hübschmann

Kleinsteuber

et al. 2012

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Modes of cytometric bacterial DNA pattern: a tool for pursuing growth

Cited by 58 publications

References 117 publications

Dynamics of Polyphosphate-Accumulating Bacteria in Wastewater Treatment Plant Microbial Communities Detected via DAPI (4′,6′-Diamidino-2-Phenylindole) and Tetracycline Labeling

Dynamics of Polyphosphate-Accumulating Bacteria in Wastewater Treatment Plant Microbial Communities Detected via DAPI (4′,6′-Diamidino-2-Phenylindole) and Tetracycline Labeling

A flow cytometric method for viability assessment of Staphylococcus aureus and Burkholderia cepacia in mixed culture

High resolution single cell analytics to follow microbial community dynamics in anaerobic ecosystems

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