Browsing by Author "Wilson, Suzanne"

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Biofilm material properties as related to shear-induced deformation and detachment phenomena
(2002-12) Stoodley, Paul; Cargo, R.; Rupp, Cory J.; Wilson, Suzanne; Klapper, Isaac
Biofilms of various Pseudomonas aeruginosa strains were grown in glass flow cells under laminar and turbulent flows. By relating the physical deformation of biofilms to variations in fluid shear, we found that the biofilms were viscoelastic fluids which behaved like elastic solids over periods of a few seconds but like linear viscous fluids over longer times. These data can be explained using concepts of associated polymeric systems, suggesting that the extracellular polymeric slime matrix determines the cohesive strength. Biofilms grown under high shear tended to form filamentous streamers while those grown under low shear formed an isotropic pattern of mound-shaped microcolonies. In some cases, sustained creep and necking in response to elevated shear resulted in a time-dependent fracture failure of the "tail" of the streamer from the attached upstream "head." In addition to structural differences, our data suggest that biofilms grown under higher shear were more strongly attached and were cohesively stronger than those grown under lower shears.
Biofilm removal from silicone tubing: an assessment of the efficacy of dialysis machine decontamination procedures using an in vitro model
(2003-01) Marion-Ferey, Karine; Pasmore, M.; Stoodley, Paul; Wilson, Suzanne; Husson, Gilles; Costerton, J. William
The aim of this study was to assess the efficacy of 21 decontamination procedures, for the removal of a multispecies biofilm. Experiments were performed on five-day-old biofilms grown inside silicone tubing, using a reactor system that mimics a dialysis machine. The treatments were tested on 5 cm tubing samples. Effects of treatment were measured using direct microscopy following staining. Bacterial viability and endotoxin removal were determined using conventional microbiological methods following biofilm detachment by scraping. The 21 procedures were classified into four groups based on the amount of biofilm removed. The most effective treatment was an acid pre-treatment, followed by use of a concentrated bleach solution. Acid pre-treatment removes calcium and magnesium carbonate crystals that are always found in dialysis biofilms. Treatments performed at high temperature did not increase the efficacy of biofilm removal. Most treatments caused at least a 105-fold reduction in bacterial viability with a few resulting in complete kill. Autoclaved and bleach-treated samples gave the best results for viability reduction, with both treatments providing an equally effective and complete kill. In addition, autoclaving led to a significant decrease in endotoxin level (removal of 99.99%).
Detachment and other dynamic processes in bacterial biofilms
(2001) Stoodley, Paul; Wilson, Suzanne; Cargo, R.; Piscitteli, C.; Rupp, Cory J.
Detachment characteristics and oxacillin resistance of Staphyloccocus aureus biofilm emboli in an in vitro catheter infection model
(2004-07) Fux, C. A.; Wilson, Suzanne; Stoodley, Paul
Catheter-related bloodstream infections due to Staphylococcus aureus are of increasing clinical importance. The pathophysiological steps leading to colonization and infection, however, are still incompletely defined. We observed growth and detachment of S. aureus biofilms in an in vitro catheter-infection model by using time-lapse microscopy. Biofilm emboli were characterized by their size and their susceptibility for oxacillin. Biofilm dispersal was found to be a dynamic process in which clumps of a wide range of diameters detach. Large detached clumps were highly tolerant to oxacillin compared with exponential-phase planktonic cultures. Interestingly, the degree of antibiotic tolerance in stationary-phase planktonic cultures was equal to that in the large clumps. The mechanical disruption of large clumps reduced the minimal bactericidal concentration (MBC) by more than 1,000 times. The MBC for whole biofilm effluent, consisting of particles with an average number of 20 bacteria was 3.5 times higher than the MBC for planktonic cultures. We conclude that the antibiotic resistance of detached biofilm particles depends on the embolus size and could be attributed to nutrient-limited stationary-phase physiology of cells within the clumps. We hypothesize that the detachment of multicellular clumps may explain the high rate of symptomatic metastatic infections seen with S. aureus.
Growth and detachment of cell clusters from mature mixed species biofilms
(2001-12) Stoodley, Paul; Wilson, Suzanne; Hall-Stoodley, Luanne; Boyle, John D.; Lappin-Scott, H. M.; Costerton, J. William
Detachment from biofilms is an important consideration in the dissemination of infection and the contamination of industrial systems but is the least-studied biofilm process. By using digital time-lapse microscopy and biofilm flow cells, we visualized localized growth and detachment of discrete cell clusters in mature mixed-species biofilms growing under steady conditions in turbulent flow in situ. The detaching biomass ranged from single cells to an aggregate with a diameter of approximately 500 µm. Direct evidence of local cell cluster detachment from the biofilms was supported by microscopic examination of filtered effluent. Single cells and small clusters detached more frequently, but larger aggregates contained a disproportionately high fraction of total detached biomass. These results have significance in the establishment of an infectious dose and public health risk assessment.
Homology of the spheroidin gene from entomopoxviruses isolated from Melanoplus sanguinipes and Amsacta moorei
(Montana State University - Bozeman, College of Agriculture, 1994) Wilson, Suzanne
The influence of fluid shear and AlCl3 on the material properties of Pseudomonas aeruginosa PAO1 and Desulfovibrio sp. EX265 biofilms
(2001) Stoodley, Paul; Jacobsen, A.; Dunsmore, B. C.; Purevdorj, B.; Wilson, Suzanne; Lappin-Scott, H. M.; Costerton, J. William
An understanding of the material properties of biofilms is important when describing how biofilms physically interact with their environment. In this study, aerobic biofilms of Pseudomonas aeruginosa PAO1 and anaerobic sulfate-reducing bacteria (SRB) biofilms of Desulfovibrio sp. EX265 were grown under different fluid shear stresses (tg) in a chemostat recycle loop. Individual biofilm microcolonies were deformed by varying the fluid wall shear stress (tw). The deformation was quantified in terms of strain (e), and the relative strength of the biofilms was assessed using an apparent elastic coefficient (Eapp) and residual strain (er) after three cycles of deformation. Aluminum chloride (AlCl3) was then added to both sets of biofilm and the tests repeated. Biofilms grown under higher shear were more rigid and had a greater yield shear stress than those grown under lower shear. The addition of AlCl3 resulted in a significant increase in Eapp and also increased the yield point. We conclude that the strength of the biofilm is in part dependent on the shear under which the biofilm was grown and that the material properties of the biofilm may be manipulated through cation cross-linking of the extracellular polysaccharide (EPS) slime matrix.
Prevention of staphylococcal biofilm-associated infections by the quorum sensing inhibitor RIP
(2005-08) Balaban, Naomi; Stoodley, Paul; Fux, C. A.; Wilson, Suzanne; Costerton, J. William; Dell'Acqua, Giorgio
Staphylococcus aureus and Staphylococcus epidermidis associated with implantable medical devices, are often difficult to treat with conventional antimicrobials. Formation of a biofilm and subsequent production of toxins are two distinct mechanisms considered important in foreign body infections. Staphylococcal virulence is caused by a complex regulatory process, which involves cell-to-cell communication through the release and response to chemical signals in a process known as quorum sensing. We explored the possibility of preventing infections by interfering with biofilm formation and toxin production using the quorum sensing inhibitor ribonucleic-acid-III-inhibiting peptide. In our studies ribonucleic-acid-III-inhibiting peptide prevented graft-associated infections caused by all species of staphylococci tested so far, including methicillin resistant S. aureus and S. epidermidis. Ribonucleic-acid-III-inhibiting peptide also enhances the effects of antibiotics and cationic peptides in the clearance of normally recalcitrant biofilm infections. Ribonucleic-acid-III-inhibiting peptide is nontoxic, highly stable, and no resistant strains have been found so far, suggesting that ribonucleic-acid-III-inhibiting peptide may be used to coat medical devices or used systemically to prevent infections. When the target of ribonucleic-acid-III activating protein activity is disrupted, biofilm formation is reduced under flow and static conditions and genes important for toxin production or biofilm formation are down-regulated. These in vitro data help explain why ribonucleic-acid-III-inhibiting peptide seems to be effective in preventing staphylococcal infections.
Statistical quantification of the detachment rates and size distribution of cell clumps from wild type (PAO1) and cell signaling mutant (JP1) Pseudomonas aeruginosa biofilms
(2004-10) Wilson, Suzanne; Hamilton, Martin A.; Hamilton, G. C.; Schumann, M. R.; Stoodley, Paul
The detachment of cells from bacterial biofilms is an important, yet poorly understood and largely unquantified phenomenon. Detached cell clumps from medical devices may form microemboli and lead to metastasis, especially if they are resistant to host defenses and antibiotics. In manufacturing plants detached clumps entering a process stream decrease product quality. Two strains of Pseudomonas aeruginosa, a wild type (PAO1) and a cell signaling mutant (JP1), were studied to (i) quantify and model detachment patterns and (ii) determine the influence of cell signaling on detachment. We collected effluent from a biofilm flowthrough reactor and determined the size distribution for cell detachment events by microscopic examination and image analysis. The two strains were similar in terms of both biofilm structure and detachment patterns. Most of the detachment events were single-cell events; however, multiple-cell detachment events contributed a large fraction of the total detached cells. The rates at which events containing multiple cells detached from the biofilm were estimated by fitting a statistical model to the size distribution data. For events consisting of at least 1,000 cells, the estimated rates were 4.5 events mm–2 min–1 for PAO1 and 4.3 events mm–2 min–1 for JP1. These rates may be significant when they are scaled up to the total area of a real biofilm-contaminated medical device surface and to the hours or days of patient exposure.