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Activity of Antibiotics against Pseudomonas aeruginosa in an In Vitro Model of Biofilms in the Context of Cystic Fibrosis: Influence of the Culture Medium.


ABSTRACT: Pseudomonas aeruginosa is a major cause of respiratory biofilm-related infections in patients with cystic fibrosis. We developed an in vitro pharmacodynamic model to study the activity of antipseudomonal antibiotics against PAO1 biofilms grown in artificial sputum medium with agar [ASM(+)] versus that against biofilms grown in Trypticase soy broth supplemented with glucose and NaCl (TGN). We measured bacterial counts, metabolic activity (fluorescein diacetate [FDA] hydrolysis), and biomass (crystal violet absorbance). Biofilms grew slower in ASM(+) than in TGN but reached the same CFU counts and metabolic activity in both media and a slightly higher biomass after 48 h in ASM(+) than in TGN. The concentration-response curves of the antibiotics after 24 h of incubation with mature biofilms showed maximal effects ranging from a 3 (ciprofloxacin)- to a 1.5 (ceftazidime, meropenem)-log10-CFU decrease, with tobramycin and colistin showing intermediate values. These maximal reductions in the numbers of CFU were similar in both media for ciprofloxacin and ?-lactams but lower in ASM(+) than in TGN for tobramycin and colistin; they were reached at concentrations lower than the human maximum concentration in plasma for ciprofloxacin and ?-lactams only. The reductions in metabolic activity and in biomass were low in both media. Small-colony variants were selected by tobramycin in ASM(+) and by ciprofloxacin in both media. The model was then successfully applied to 4 isolates from patients with cystic fibrosis. These biofilms showed CFU counts similar to those of PAO1 biofilms in ASM(+) but a higher biomass than PAO1 biofilms in ASM(+) and moderate differences in their susceptibility to antibiotics from that of PAO1 biofilms grown in this medium. This model proved useful to establish the pharmacodynamic profile of drugs against P. aeruginosa biofilms in the context of cystic fibrosis.

SUBMITTER: Diaz Iglesias Y 

PROVIDER: S-EPMC7179293 | BioStudies | 2020-01-01

REPOSITORIES: biostudies

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