Determining the spatial distribution of viable and nonviable bacteria in hydrated microcosm dental plaques by viability profiling
Open Access
- 1 September 2002
- journal article
- research article
- Published by Oxford University Press (OUP) in Journal of Applied Microbiology
- Vol. 93 (3), 448-455
- https://doi.org/10.1046/j.1365-2672.2002.01703.x
Abstract
Aims: The aim of this study was to use confocal laser scanning microscopy (CLSM) to examine the spatial distribution of both viable and nonviable bacteria within microcosm dental plaques grown in vitro. Previous in vivo studies have reported upon the distribution of viable bacteria only. Methods and Results: Oral biofilms were grown on hydroxyapatite (HA) discs in a constant‐depth film fermenter (CDFF) from a saliva inoculum. The biofilms were stained with the BacLightTM LIVE/DEAD system and examined by CLSM. Fluorescence intensity profiles through the depth of the biofilm showed an offset between the maximum viable intensity and the maximum nonviable intensity. Topographical differences between the surface properties of the viable and nonviable biofilm virtual surfaces were also measured. Conclusions: The profile of fluorescence intensity from viable and nonviable staining suggested that the upper layers of the biofilm contain proportionally more viable bacteria than the lower regions of the biofilm. Significance and Impact of Study: Viability profiling records the transition from predominantly viable to nonviable bacteria through biofilms suggesting that this technique may be of use for quantifying the effects of antimicrobial compounds upon biofilms. The distribution of viable bacteria was similar to that found in dental plaque in vivo suggesting that the CDFF produces in vitro biofilms which are comparable to their in vivo counterparts in terms of the spatial distribution of viable bacteria.Keywords
This publication has 14 references indexed in Scilit:
- Spatial distribution of vital and dead microorganisms in dental biofilmsArchives of Oral Biology, 2001
- Heterogeneity in biofilmsFEMS Microbiology Reviews, 2000
- Structural studies of microcosm dental plaques grown under different nutritional conditionsFEMS Microbiology Letters, 2000
- Transfer of a conjugative transposon, Tn5397 in a model oral biofilmFEMS Microbiology Letters, 1999
- A pilot study of confocal laser scanning microscopy for the assessment of undisturbed dental plaque vitality and topographyArchives of Oral Biology, 1998
- Development of a steady-state oral microbial biof ilrn community using the constant-depth film ferrnenterMicrobiology, 1996
- Corrosion of intra-oral magnets in the presence and absence of biofilms of Streptococcus sanguisBiomaterials, 1995
- A constant‐depth laboratory model film fermentorBiotechnology & Bioengineering, 1988
- Structure of the Microbial Flora Associated with Periodontal Health and Disease in Man: A Light and Electron Microscopic StudyThe Journal of Periodontology, 1976
- Ultrastructure of the attachment device between coccal and filamentous microorganisms in “corn cob” formations of dental plaqueArchives of Oral Biology, 1973