Document Type thesis Author Name Tao, Yuanyuan URN etd-122010-152311 Title Effects of Cranberry Juice Cocktail on Surface Adhesion and Biofilm Formation of Uropathogenic Bacteria Degree MS Department Chemical Engineering Advisors Terri A. Camesano, Advisor David DiBiasio, Department Head Keywords urinary tract infections Escherichia coli atomic force microscopy Vacciniumm macrocarpon Date of Presentation/Defense 2010-12-20 Availability unrestricted
American cranberry (Vacciniumm macrocarpon) has been long known for its benefits in maintaining urinary tract health. Clinical trials have shown that drinking cranberry juice can prevent urinary tract infections (UTIs) in various subpopulations that are prone to UTIs, especially women, but the mechanisms by which cranberry acts against uropathogenic bacteria are still unclear. Studies showed that when exposed to cranberry juice or A- PACs, a group of tannins that are unique to cranberry, the adhesion activity and biofilm formation of uropathogenic bacteria were reduced. However, the metabolism of cranberry juice has not be elucidated, therefore further study is needed to find out whether the anti-bacterial components in cranberry could survive the digestive system and reach the urinary tract, and how the components or metabolites remaining in urine act against uropathogenic bacteria. We used atomic force microscopy (AFM) to study the surface adhesion force of uropathogenic E. coli incubated with urine samples that were collected from volunteers after drinking 16 oz. of cranberry juice cocktail (CJC) or water. The urine samples were collected at 0, 2, 4, 6, and 8 hours after CJC or water consumption. When incubated with post-water urine, the adhesion forces of pathogenic bacteria that have fimbriae (E. coli B37, B73, B78, BF1023, CFT 073, and J96) did not change; whereas the adhesion forces of these strains decreased over the 8 hour period after CJC consumption. The control strain that does not have frimbriae, E. coli HB101, showed low adhesion force when incubated with post-water and post-CJC urine. In a human red blood cell agglutination (HRBC) assay, the attachment of pathogenic E. coli to red blood cells was significantly lower after exposed to post-CJC urine, compared to those exposed to post-water urine. These results indicate the anti-bacteria components or metabolites of CJC stay active in urine, and these compounds prevent adhesion of E. coli by reducing fimbriae-mediated adhesion. We also examined the effects of drinking CJC on biofilm formation of uropathogenic bacteria. Female volunteers were given 16 oz. of CJC or placebo, and their urine was collected at 0, 2, 8, 24, and 48 hours after consumption. Bacteria (E. coli B37, CFT073, BF1023, HB101, and S. aureus ATCC43866) were cultured in a mixture of urine and growth media in 96 well microtiters. The biofilm formed was quantified by staining the biofilm dissolved in a solvent with crystal violet and measuring the absorbance at 600 nm. The results showed that biofilm formation was reduced within 24 hours after CJC consumption, and it started to increase after 48 hours, possibly due to the washout of CJC in the system. These studies suggest that CJC can be an effective preventive measure for UTIs as it inhibits adhesion and biofilm formation of uropathogenic bacteria.
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