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. 1995 Dec;61(12):4357–4361. doi: 10.1128/aem.61.12.4357-4361.1995

Direct and Rapid Analysis of the Adhesion of Bacteria to Solid Surfaces: Interaction of Fluorescently Labeled Rhodococcus Strain GIN-1 (NCIMB 40340) Cells with Titanium-Rich Particles

G Fleminger, Y Shabtai
PMCID: PMC1388653  PMID: 16535188

Abstract

A fluorimetric assay which enables direct and accurate analysis of the adhesion of bacteria to solid particles was developed. The assay is based on labeling of the bacteria with fluorescamine, which reacts with primary amino groups on the cell surface to yield a yellow fluorescence that is easily detectable by both fluorescence microscopy and spectrofluorimetry. As an example, fluorescent labeling of Rhodococcus strain GIN-1 (NCIMB 40340) cells enabled the detection and quantitative determination of their adsorption to TiO(inf2) and coal fly ash particles. Exposure of the cells to 10% acetone during the labeling reaction affected neither their viability nor their ability to adhere to these particles. Only a small fraction (^sim2%) of the total cell protein was labeled by fluorescamine upon staining of intact bacterial cells, which may indicate preferential labeling of certain proteins. Specificity studies carried out with the fluorescence assay confirmed previous findings that Rhodococcus strain GIN-1 cells possess high affinities for TiO(inf2), ZnO, and coal fly ash and low affinities for other metal oxides. In principle, the newly developed fluorimetric assay may be used for determination of cell adhesion to any solid matrix by either microscopic examination or epifluorescence measurements. In the present work, the adhesion of several other microorganisms to TiO(inf2) particles was tested as well, but their ability to adhere to these particles was significantly lower than that of Rhodococcus strain GIN-1 cells.

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Selected References

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  1. Amann R. I., Stromley J., Devereux R., Key R., Stahl D. A. Molecular and microscopic identification of sulfate-reducing bacteria in multispecies biofilms. Appl Environ Microbiol. 1992 Feb;58(2):614–623. doi: 10.1128/aem.58.2.614-623.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ellen R. P., Schwarz-Faulkner S., Grove D. A. Coaggregation among periodontal pathogens, emphasizing Bacteroides gingivalis--Actinomyces viscosus cohesion on a saliva-coated mineral surface. Can J Microbiol. 1988 Mar;34(3):299–306. doi: 10.1139/m88-055. [DOI] [PubMed] [Google Scholar]
  3. Hussain M., Collins C., Hastings J. G., White P. J. Radiochemical assay to measure the biofilm produced by coagulase-negative staphylococci on solid surfaces and its use to quantitate the effects of various antibacterial compounds on the formation of the biofilm. J Med Microbiol. 1992 Jul;37(1):62–69. doi: 10.1099/00222615-37-1-62. [DOI] [PubMed] [Google Scholar]
  4. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  5. Marshall K. C., Cruickshank R. H. Cell surface hydrophobicity and the orientation of certain bacteria at interfaces. Arch Mikrobiol. 1973 Apr 8;91(1):29–40. doi: 10.1007/BF00409536. [DOI] [PubMed] [Google Scholar]
  6. Rogers J., Keevil C. W. Immunogold and fluorescein immunolabelling of Legionella pneumophila within an aquatic biofilm visualized by using episcopic differential interference contrast microscopy. Appl Environ Microbiol. 1992 Jul;58(7):2326–2330. doi: 10.1128/aem.58.7.2326-2330.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Shabtai Y., Fleminger G. Adsorption of Rhodococcus Strain GIN-1 (NCIMB 40340) on Titanium Dioxide and Coal Fly Ash Particles. Appl Environ Microbiol. 1994 Sep;60(9):3079–3088. doi: 10.1128/aem.60.9.3079-3088.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Stein S., Böhlen P., Udenfriend S. Studies on the kinetics of reaction and hydrolysis of fluorescamine. Arch Biochem Biophys. 1974 Jul;163(1):400–403. doi: 10.1016/0003-9861(74)90491-3. [DOI] [PubMed] [Google Scholar]
  9. Udenfriend S., Stein S., Böhlen P., Dairman W., Leimgruber W., Weigele M. Fluorescamine: a reagent for assay of amino acids, peptides, proteins, and primary amines in the picomole range. Science. 1972 Nov 24;178(4063):871–872. doi: 10.1126/science.178.4063.871. [DOI] [PubMed] [Google Scholar]
  10. Williamson F. A., Palframan K. R. An improved method for collecting and staining microorganisms for enumeration by fluorescence light microscopy. J Microsc. 1989 Jun;154(Pt 3):267–272. doi: 10.1111/j.1365-2818.1989.tb00589.x. [DOI] [PubMed] [Google Scholar]

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