The adhesion of bacteria to uroepithelial cells or urinary catheters is the first step in the development of biofilm formation and urinary tract infections (UTIs). Previous research has suggested that consumption of cranberry juice can prevent the recurrence of UTIs by decreasing bacterial adhesion since isolated compounds in cranberries, known as A-type proanthocyanidins (PACs), change the conformation of proteinaceous fimbriae that help attach bacteria to epithelial cell receptors. Most clinical and laboratory studies have shown the effects of cranberry juice cocktail (CJC) on large communities of bacteria; however, very few studies have evaluated how cranberry affects the adhesion forces of a single bacterium as well as effects on cellular composition and biofilm formation. We used atomic force microscopy (AFM) to investigate the effects of CJC and PACs on the adhesion forces between E. coli and a silicon nitride tip. Bacterial cultures were grown in tryptic soy broth (TSB), supplemented with 0 and 10 wt.% light cranberry juice cocktail (L-CJC) or 128 µg/mL PACs. E. coli bacteria were continuously cultured in the presence of cranberry products up to twelve times. Experiments were conducted at different scales to test bacterial attachment and adhesion forces. At the macroscale, bacteria were incubated with uroepithelial cells and the number of bacteria attached per uroepithelial cell was determined. In nanoscale experiments, the forces of adhesion between E. coli and a silicon nitride AFM tip were probed for bacteria grown in L-CJC or PACs for different numbers of culture times. Successive replacement of media and continued culture in L-CJC and PACs resulted in a significant decrease in adhesion forces for E. coli strains. Finally, during the continuous exposure of L-CJC to bacteria we examined the growth, morphology, and ability to form biofilms of E. coli. We found a decrease in growth rates related to changes in Gram staining with increasing number of cultures in L-CJC. Growth of bacteria in L-CJC or PACs inhibited the development of biofilms on polyvinyl-chloride, which can model biofilm formation on urinary catheters. We also determined that growth of E. coli in L-CJC results in prevention of the expression of indole which can be linked to the inhibition of biofilm formation. Our results help support the molecular mechanisms for the role of cranberry in preventing the adhesion of E. coli to biotic and abiotic surfaces, thus helping to scientifically validate the use of cranberry juice as a prophylactic treatment for the prevention of UTIs.
| Identifer | oai:union.ndltd.org:wpi.edu/oai:digitalcommons.wpi.edu:etd-theses-1041 |
| Date | 09 January 2008 |
| Creators | Pinzon-Arango, Paola A. |
| Contributors | Terri A. Camesano, Advisor, George D. Pins, Committee Member, Reeta Prusty Rao, Committee Member |
| Publisher | Digital WPI |
| Source Sets | Worcester Polytechnic Institute |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | Masters Theses (All Theses, All Years) |
Page generated in 0.0023 seconds