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Folia Microbiologica

, Volume 64, Issue 4, pp 567–577 | Cite as

Quantitative evaluation of biofilm extracellular DNA by fluorescence-based techniques

  • Martina BoháčováEmail author
  • Jarmila Pazlarová
  • Viviana Fuchsová
  • Tereza Švehláková
  • Kateřina Demnerová
Original Article
  • 297 Downloads

Abstract

The formation of a hardly removable biofilm in food processing and clinical settings calls for a deeper understanding of composition of the matrix that protects the biofilm cells, as the crucial matrix component is extracellular DNA (eDNA), participating in adhesion, aggregation and penetration reduction, yet serving as a horizontal gene transfer reservoir. Therefore, we evaluated eDNA release from the biofilm of two pathogens, Listeria monocytogenes and Staphylococcus aureus, with respect to their origin under different culturing condition. Primarily, the biofilms were observed by confocal laser scanning microscopy (CLSM) under conditions mimicking the food processing environment and human body. The eDNA was quantitatively characterised based on its area by IMARIS. Next, the eDNA content and biofilm formation were quantified by spectrophotometry. Data from both sets of experiments were statistically evaluated. The eDNA release varied between the microorganism, culturing conditions and the origin of strains. Independent of the method used, the clinical strains of S. aureus released more eDNA than the food related strains at 37 °C. eDNA content can be crucial discriminating matrix component between food related and clinical strains. Deeper understanding of the eDNA role in such a phenomenon could facilitate the design of effective strategy for biofilm disruption.

Notes

Acknowledgements

We would like to acknowledge data processing consultations with Ing. Robert Šulák.

Funding information

This work received financial support from specific university research MSMT No. 20-SVV/2016 and the Czech Science Foundation GAČR 17-15936S.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

12223_2019_681_MOESM1_ESM.docx (15 kb)
ESM 1 (DOCX 15 kb)

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Copyright information

© Institute of Microbiology, Academy of Sciences of the Czech Republic, v.v.i. 2019

Authors and Affiliations

  1. 1.Faculty of Food and Biochemical TechnologyUniversity of Chemistry and Technology, PraguePragueCzech Republic

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