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Cytochemical Differences in Bacterial Glycocalyx

Published online by Cambridge University Press:  28 January 2005

Wolf Dietrich Krautgartner ,
Ljubomir Vitkov ,
Matthias Hannig ,
Klaus Pelz  and
Walter Stoiber
Wolf Dietrich Krautgartner
Affiliation:
Department of Electron Microscopy, Light Microscopy and Digital Image Acquisition, Institute of Zoology, University of Salzburg, Hellbrunnerstraße 34, 5020 Salzburg, Austria
Ljubomir Vitkov
Affiliation:
Department of Electron Microscopy, Light Microscopy and Digital Image Acquisition, Institute of Zoology, University of Salzburg, Hellbrunnerstraße 34, 5020 Salzburg, Austria Department of Operative Dentistry and Periodontology, Saarland University, D-66421 Homburg/Saar, Germany
Matthias Hannig
Affiliation:
Department of Operative Dentistry and Periodontology, Saarland University, D-66421 Homburg/Saar, Germany
Klaus Pelz
Affiliation:
Institute of Medical Microbiology and Hygiene, University of Freiburg, D-79104 Freiburg, Germany
Walter Stoiber
Affiliation:
Department of Electron Microscopy, Light Microscopy and Digital Image Acquisition, Institute of Zoology, University of Salzburg, Hellbrunnerstraße 34, 5020 Salzburg, Austria
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Abstract

To examine new cytochemical aspects of the bacterial adhesion, a strain 41452/01 of the oral commensal Streptococcus sanguis and a wild strain of Staphylococcus aureus were grown with and without sucrose supplementation for 6 days. Osmiumtetraoxyde (OsO4), uranyl acetate (UA), ruthenium red (RR), cupromeronic blue (CB) staining with critical electrolytic concentrations (CECs), and the tannic acid–metal salt technique (TAMST) were applied for electron microscopy. Cytochemically, only RR-positive fimbriae in S. sanguis were visualized. By contrast, some types of fimbriae staining were observed in S. aureus glycocalyx: RR-positive, OsO4-positive, tannophilic and CB-positive with ceasing point at 0.3 M MgCl2. The CB staining with CEC, used for the first time for visualization of glycoproteins of bacterial glycocalyx, also reveals intacellular CB-positive substances—probably the monomeric molecules, that is, subunits forming the fimbriae via extracellular assembly. Thus, glycosylated components of the biofilm matrix can be reliably related to single cells. The visualization of intracellular components by CB with CEC enables clear distinction between S. aureus and other bacteria, which do not produce CB-positive substances. The small quantities of tannophilic substances found in S. aureus makes the use of TAMST for the same purpose difficult. The present work protocol enables, for the first time, a partial cytochemical differentiation of the bacterial glycocalyx.

Keywords

fimbriaeruthenium redcupromeronic bluecritical electrolytic concentrationstannic acid–metal salt techniqueStaphylococcus aureusStreptococcus sanguis
Type
BIOLOGICAL APPLICATIONS
Information
Microscopy and Microanalysis , Volume 11 , Issue 1 , February 2005 , pp. 2 - 8
Copyright
© 2005 Microscopy Society of America

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