A Rapid Bioluminescence Method for Quantifying Bacterial Adhesion to Polystyrene

Harber, M. J.; Mackenzie, Ruth; Asscher, A. W.

doi:10.1099/00221287-129-3-621

Cited by 18 publications

(9 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…et al, 1986), by quantitative culture (Sheth et al, 1983;Mackenzie and Rivera-Calderon, 1985), by Discussion Bacterial rials can be use of radiolabelled bacteria or by assay of ATP by bioluminescence (Harber et al, 1983). Each method has some disadvantages, but ATP bioluminescence provides a rapid and simple quantitative assay of bacterial biomass biomass adherent to prosthetic matemeasured by direct microscopy (Hogt attached to synthetic polymer surfaces (Ludwicka et al, 1985).…”

Section: Scanning Electronmicroscopymentioning

confidence: 99%

Adherence of staphylococci to intravascular catheters

Kristinsson

1989

Journal of Medical Microbiology

View full text Add to dashboard Cite

Summary. Adherence of seven strains of Staphylococcus epidermidis and three strains of S . aureus to three types of intravascular catheters was assessed by ATP bioluminescence, by culture after ultrasonication and by scanning electronmicroscopy. The catheter materials studied were silicone elastomer, thermoplastic polyurethane and polyurethane coated with Hydromer@, a coating which absorbs water and provides a hydrophilic sheath around the catheter. The adherence assays were performed in phosphate-buffered saline on a rotary shaker at 37"C, with the catheters precoated with serum and uncoated, and the results were correlated with bacterial hydrophobicity. There was wide strain-to-strain variation in bacterial adherence ; S . aureus and slime-producing S . epidermidis strains adhered better than did non-slimeproducing strains. Overall, there was less bacterial adherence to Hydromer@-coated catheters than to polyurethane and silicone catheters but it was unrelated to bacterial hydrophobicity. Serum coating of catheters resulted in marked reduction of bacterial adherence.

show abstract

Section: Scanning Electronmicroscopymentioning

confidence: 99%

Adherence of staphylococci to intravascular catheters

Kristinsson

1989

Journal of Medical Microbiology

View full text Add to dashboard Cite

show abstract

“…Factors which influence attachment include those which influence the physicochemistry of adhesion, such as electrolyte species or concentration (10,15,17), pH (17,23), temperature (4,11), and surface (3,20) and medium (6,16) composition. Also as important, but more difficult to elucidate, are factors which influence physiological processes related to adhesion (5,18), such as exopolymer production (24).…”

mentioning

confidence: 99%

Attachment of Pseudomonas fluorescens to glass and influence of electrolytes on bacterium-substratum separation distance

Fletcher

1988

J Bacteriol

140

View full text Add to dashboard Cite

The influence of Na+, Ca2+, La3+, and Fe3+ on the adhesion of Pseudomonasfluorescens H2 and H2S was investigated with interference reflection microscopy (IRM). IRM is a light microscopy technique which allows (i) visualization of the adhesive sites of living bacteria as they attach to a glass cover slip surface and (ii) evaluation of the bacterium-glass surface separation distance within a range of 0 to ca. 100 nm. The addition of each cation caused changes in IRM images consistent with a decrease in the separation distance, and minimum effective concentrations were as follows: Na+, 1 mM; Ca2+, 1 mM; La3+, 50 I,M; and Fe3+, 50 ,uM.With strain H2, the effects of Na+, Ca2+, and La3+ were fully reversible in that the separation distance increased again when the electrolyte was replaced with distilled water. However, with strain H2S, a spontaneous mutant of H2 with increased attachment ability, only the effect of Na+ was fully reversible, and the effects of Ca2' and La3+ were only partially reversible or irreversible. The effect of Fe3+ was irreversible with both strains, but this may be related not only to the electrolytic nature of Fe3+ but also to the decrease in solution pH to 3.5 caused by its addition. It is proposed that the electrolytes caused a decrease in separation distance by neutralizing negative charges on bacterial surface polymers and that the different effects obtained with the two strains are related to their different adhesion abilities.When bacteria in an aqueous suspension are exposed to a solid surface, many of those bacteria may adhere to the surface and become permanently attached. Investigators of bacterial attachment have attempted to determine the adhesion mechanism(s) of different bacteria or to test the effects of different environmental or physiological conditions on attachment (14). However, there has been much variation in results from these studies with respect to the ability of different bacterial species, strains, or phenotypes to attach and the susceptibility of their adhesion to modifying factors. Because of this variation, it has been extremely difficult to determine the mechanism by which a given factor can modify the adhesion process.Factors which influence attachment include those which influence the physicochemistry of adhesion, such as electrolyte species or concentration (10,15,17), pH (17, 23), temperature (4, 11), and surface (3,20) Meaningful direct observation of attaching cells has been impossible because of the limiting resolution of light microscopy and the inability to view the bacterial surface-substratum interface during adhesion.However, with interference reflection microscopy (IRM), the adhesive sites on the bacterial surface can be visualized while attachment to a solid surface is established. With this technique, it is possible to observe living cells as they become loosely associated (i.e., within 100 nm) with a glass cover slip surface and to evaluate the change in distance between the cell and surface as firm adhesion is established. With IRM, the darknes...

show abstract

“…Bacterial cells from the third subculture were harvested by centrifugation, washed twice in PBS and resuspended in this buffer to an optical density at 560 nm of 2.0 which gave about 2 X lo9 colony forming units (cfu) per ml. Production of type 1 fimbriae by E.coli 504 was confirmed by transmission electron microscopy and mannose-sensitive haemagglutination of guinea pig erythrocytes (Harber et al, 1983).…”

Section: Stimulimentioning

confidence: 99%

Factors affecting the measurement of chemiluminescence in stimulated human polymorphonuclear leucocytes

Harber

Topley

1986

J. Biolumin. Chemilumin.

View full text Add to dashboard Cite

Optimum conditions were established for the generation and measurement of luminol-dependent chemiluminescence (CL) in human polymorphonuclear leucocytes (PMNL) stimulated with a variety of particulate and soluble agents. Several factors had a particular influence on the kinetics of CL stimulated by the chemotactic peptide N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP). Two peaks, both azide-sensitive, were observed at 21 degrees C and 25 degrees C, but these increased in magnitude and merged to give a single, early peak when the temperature was increased at 37 degrees C. Pre-exposure of PMNL to a buffer containing calcium was essential for the expression of both phases of fMLP-stimulated CL, while the second peak decreased dramatically if the cells were stored at 4 degrees C for 4 hours before assay. In contrast, storage of PMNL at 4 degrees C for up to 8 hours in a buffer without divalent cations did not alter the kinetics or magnitude of CL induced by other stimuli, and had the benefit of minimizing the rate of cell aggregation. This study confirms that measurement of luminol-dependent CL in stimulated PMNL is a useful analytical tool, but shows that careful attention to experimental design is required to ensure that the observed CL provides a true measure of the parameter under investigation.

show abstract

A Rapid Bioluminescence Method for Quantifying Bacterial Adhesion to Polystyrene

Cited by 18 publications

References 26 publications

Adherence of staphylococci to intravascular catheters

Adherence of staphylococci to intravascular catheters

Attachment of Pseudomonas fluorescens to glass and influence of electrolytes on bacterium-substratum separation distance

Factors affecting the measurement of chemiluminescence in stimulated human polymorphonuclear leucocytes

Contact Info

Product

Resources

About