RESUMEN
BACKGROUND: The known bactericidal properties of ozone have not been checked in relation to its action on bacterial biofilms. This is especially true of ozonated fluids. The aim of this study was to investigate the bactericidal activity of ozonated water and that of a mixture of ozone and oxygen against biofilms. MATERIAL/METHODS: Eighteen clinical strains of Staphylococcus aureus and Pseudomonas aeruginosa exhibiting various levels of antibiotic sensitivity were investigated. Bacteria were cultured in biofilm form on polystyrene titration plates for periods of 2 to 72 hours. The biofilms formed in this way were exposed to in statu nascendi ozonated water produced in a prototype device that had been tested in clinical conditions, or to a mixture of oxygen and ozone generated in the same device. Live cells in the biofilm were stained with a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) bromide solution. The degree of reduction of viable bacteria following ozone exposure was determined. RESULTS: Ozonated water was found to be an effective bactericidal agent against biofilms after as little as 30 seconds of exposure, while the bactericidal activity of the ozone-oxygen solution was much lower. Prolongation of the duration of biofilm exposure to the gaseous disinfectant to 40 minutes led to a reduction in the viable cell count, which nevertheless remained high. CONCLUSIONS: Unlike the ozone-oxygen mixture, ozonated water effectively destroys bacterial biofilms in vitro.
Asunto(s)
Biopelículas/efectos de los fármacos , Ozono/farmacología , Pseudomonas aeruginosa/efectos de los fármacos , Staphylococcus aureus/efectos de los fármacos , Oxígeno/química , Ozono/química , Análisis de Supervivencia , Sales de Tetrazolio , Tiazoles , Agua/químicaRESUMEN
The aim of this study was to analyse P aeruginosa biofilm growing in three media, using three methods of biofilm staining: with 2% crystal violet solution (CV), with 1% 2,3,5-triphenyltetrazolium chloride solution (TTC) and 3-(4,5-dimethyl-2-thiazolyl)-2,5 diphenyl-2H-tetrazolium bromide (MTT). Using the method with MTT solution we could observe growth of strains forming biofilm both slowly or quickly. Achieved values of absorbation allow to observe dynamic of growing selected strains in the period of time. Among three groups of analysed strains, the biggest percentage of medium growing strains were achieved, while in the fourth group (strains isolated from spit of patients with cystic fibrosis) the biggest percentage of low growing strains was observed. Achieved results allow to select the strains quickly forming biofilm, the best media (LB, TSB2%glu) and method of biofilm staining (MTT), which will be used in the further studies.