High-resolution novel method for tracking bacteria in a multi-species biofilm

David Polak*, Sharon Shany-Kdoshim, Liana Zaydel, Osnat Feuerstein, Yael Houri-Haddad

*Corresponding author for this work

    Research output: Contribution to journalArticlepeer-review

    6 Scopus citations

    Abstract

    The aim of this study is to establish a novel high resolution tracking ability of a specific bacterium in multispecies biofilm. A periodontal multispecies biofilm was constructed with Streptococcus sanguis, Actinomyces naeslundii, Porphyromonas gingivalis and Fusobacterium nucleatum. A single species was stained with fluorescein isothiocyanate (FITC). The mature biofilm was stained for viability (propidium iodide) and analysis was performed with flow cytometry. The sensitivity of the assay was compared with colony forming units (CFU) counts. A single cell suspension of P. gingivalis was grown in broth and biofilm to identify the location of these events on side scatter and forward scatter. The sensitivity of the assay was comparable to that of the CFU counts. The assay allows quantification of the ratio of a single bacterium within the biofilm, and its viable proportion. The described method is reproducible and of high resolution, and allows the examination of microbes’ composition and viability within a biofilm structure.

    Original languageAmerican English
    Pages (from-to)259-266
    Number of pages8
    JournalArchives of Microbiology
    Volume201
    Issue number2
    DOIs
    StatePublished - 2 Mar 2019

    Bibliographical note

    Publisher Copyright:
    © 2019, Springer-Verlag GmbH Germany, part of Springer Nature.

    Keywords

    • Bacterial viability
    • Flow cytometry
    • Multispecies biofilm
    • Periodontitis

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