title: Effects of single- and multi-strain probiotics on biofilm formation and in vitro adhesion to bladder cells by urinary tract pathogens.
abstract: OBJECTIVE
  There is increasing evidence that probiotic bacteria can inhibit and/or prevent urinary tract infections. Possible mechanisms include prevention of adhesion of pathogens to the bladder epithelium and inhibition of biofilm formation. Currently there is interest in the comparative efficacy of single probiotics vs. strain mixtures. We have therefore tested the inhibitory activity of four single probiotics and four probiotic mixtures towards the urinary tract pathogens Escherichia coli NCTC 9001 and Enterococcus faecalis NCTC 00775.
  
  
  METHODS
  Inhibition of biofilm formation by cell-free supernatants was tested using the Crystal Violet assay, while prevention of pathogen adhesion to host cells was tested by using bladder cancer cells as a model for the human urinary tract.
  
  
  RESULTS
  Under pH-controlled conditions, there was no significant inhibition of biofilm formation by any treatment. Without pH control, 5/8 treatments significantly inhibited biofilm production by E. coli, while 5/8 treatments inhibited production by E. faecalis. Using data from all Crystal Violet assays, there was no significant difference in the ability of single- and multi-strain probiotics to inhibit biofilm formation. In the cell culture assays, all treatments were able to significantly reduce numbers of pathogenic cells adhering to host cells by 2.5-3.5 logs. No significant difference was observed between the displacement caused by single strains and mixtures for either pathogen.
  
  
  CONCLUSIONS
  Inhibition of biofilm seems to be a major mechanism of urinary tract pathogen exclusion, related to, and possibly dependent upon, the probiotic ability to reduce environmental pH. Exclusion via competition of binding sites is a possible in vivo mechanism for these probiotics. If an additive or synergistic effect exists between strains within a mixture, it does not manifest itself in a greater effect through these two inhibitory mechanisms.
authors.affiliation: Department of Food and Nutritional Sciences, University of Reading, P.O. Box 226, Whiteknights, Reading RG6 6AP, United Kingdom. Electronic address: chapmancmc@hotmail.com.
authors.forename: C M C
authors.initials: CM
authors.lastname: Chapman
authors.affiliation: Department of Food and Nutritional Sciences, University of Reading, P.O. Box 226, Whiteknights, Reading RG6 6AP, United Kingdom.
authors.forename: G R
authors.initials: GR
authors.lastname: Gibson
authors.affiliation: Department of Food and Nutritional Sciences, University of Reading, P.O. Box 226, Whiteknights, Reading RG6 6AP, United Kingdom.
authors.forename: I
authors.initials: I
authors.lastname: Rowland
mesh_terms: D000898:Antibiosis; D001422:Bacterial Adhesion; D018441:Biofilms / Q000254:growth & development*; D045744:Cell Line, Tumor; D013293:Enterococcus faecalis / Q000502:physiology*; D004847:Epithelial Cells / Q000382:microbiology*; D004926:Escherichia coli / Q000502:physiology*; D006801:Humans; D019936:Probiotics / Q000494:pharmacology*; D001743:Urinary Bladder / Q000382:microbiology*
keywords: Cell culture; Crystal Violet; Multi-species; Probiotic; Urinary