Enhanced activity of combination of tobramycin and piperacillin for eradication of sessile biofilm cells of Pseudomonas aeruginosa

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Abstract
An in vitro chemostat system in which Pseudomonas aeruginosa can be cultivated at a slow growth rate and under iron limitation conditions was used to study the susceptibilities of sessile bacteria of mucoid and nonmucoid P. aeruginosa strains to tobramycin and piperacillin. Planktonic cells of both mucoid and nonmucoid P. aeruginosa strains were susceptible to tobramycin and piperacillin. None of the cells was found to be viable after 2 h of exposure to 200 micrograms of piperacillin plus 10 micrograms of tobramycin per ml. Young sessile bacteria were slightly more resistant to piperacillin or tobramycin than the planktonic cells were. However, eradication of young sessile bacteria could be achieved with a combination of piperacillin and tobramycin. None of these young biofilm bacteria were found to be viable after a 2-h exposure to 200 micrograms of piperacillin plus 10 micrograms of tobramycin per ml. Old sessile bacteria were very resistant to these antibiotics. Eradication of old sessile bacteria could not be achieved with either tobramycin (200 micrograms/ml) or piperacillin (200 micrograms/ml) alone. Combination of higher concentrations of tobramycin with piperacillin resulted in an enhancement of killing of the old sessile bacteria. Exposure of old sessile bacteria to 200 micrograms of piperacillin plus 100 micrograms of tobramycin per ml resulted in the reduction of the viable count to approximately 0.02%. The data suggest that the eradication of biofilm-associated infections is best carried out as early as possible. Enhanced activities against the sessile bacteria were achieved when higher concentrations of aminoglycosides were combined with beta-lactam antibiotics.