title: Biofilm culture of Pseudomonas aeruginosa expressing lux genes as a model to study susceptibility to antimicrobials.
abstract: A simple in vitro model for culture of biofilm populations of self-bioluminescent Pseudomonas aeruginosa was used for real-time monitoring of the effects of ciprofloxacin. Biofilms of these organisms were established within Sorbarod filters, perfused with a chemically defined simple salts medium. The biofilm population was shown to achieve a pseudo-steady state which was reproducible and stable over several days. The viability of membrane-associated and eluted cells was assessed by spread plate viable counts and by monitoring bioluminescence as a measure of metabolic activity. Pseudo-steady state biofilms were exposed to 5x MIC ciprofloxacin (0.3 mg x l(-1)) in the perfusing medium for 1 h. Whilst both methods for viability assessment indicated an immediate reduction in viable cell numbers, the decline recorded with bioluminescence was greater. The use of bioluminescent bacteria proved to be a rapid and sensitive method for the measurement of real-time antibacterial effects on a bacterial biofilm.
authors.affiliation: Department of Biological and Biomedical Science, University of the West of England, BS16 1QT, Bristol, UK.
authors.forename: A
authors.initials: A
authors.lastname: Parveen
authors.forename: G
authors.initials: G
authors.lastname: Smith
authors.forename: V
authors.initials: V
authors.lastname: Salisbury
authors.forename: S M
authors.initials: SM
authors.lastname: Nelson
mesh_terms: D000890:Anti-Infective Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D018441:Biofilms / Q000187:drug effects* / Q000254:growth & development; D002939:Ciprofloxacin / Q000494:pharmacology*; D003470:Culture Media; D008163:Luminescent Measurements; D008826:Microbial Sensitivity Tests / Q000379:methods; D011550:Pseudomonas aeruginosa / Q000187:drug effects* / Q000235:genetics / Q000502:physiology*
chemical_list: D000890:Anti-Infective Agents; D001426:Bacterial Proteins; D003470:Culture Media; D002939:Ciprofloxacin