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In vitro Investigation of the MexAB Efflux Pump From Pseudomonas aeruginosa
Published on: February 17, 2014
Capillary electrophoresis study of outer membrane proteins of Pseudomonas strains upon antibiotic treatment
Tamás Kustos1, Ildikó Kustos, Edit Gonda
1Department of Orthopaedics, Faculty of Medicine, University of Pécs, Szigeti út 12, H-7643 Pécs, Hungary.
Abstract:
Nosocomial wound infections by Pseudomonas aeruginosa strains have increasing importance in orthopaedic surgery. Outer membrane protein composition and cell-surface hydrophobicity of the bacteria have strong influence on adhesion to living tissues or artificial medical devices. Outer membrane proteins of five Pseudomonas strains (KT 2, KT 7, KT 25, KT 28, KT 39) isolated from orthopaedic patients' wounds and one standard strain NIH Hungary 170000 isolated from pus were examined. The capillary electrophoretic patterns of the outer membrane proteins were characteristic to each bacterial strains possessing different relative ratios of major and minor proteins. Antibiotic treatment of bacteria with three antibiotics, cefotaximum, amoxicillinum/clavulinic acid and amikacinum (applied often in prophylaxis and treatment of patients) induced changes in the electrophoretic profiles showing that outer membrane protein composition was altered significantly. The most pronounced alterations in the electrophoretic patterns after antibiotic treatment were obtained in the cases of the strains KT 2, KT 7 and KT 28. The amikacinum administration strongly decreased the relative percentage of the 38800 rel. mol. mass protein in KT 2 (from 20 to 6%). while the relative amount of the same protein increased significantly in KT 7 and KT 28 after cefotaximum treatment (from 2 to 16% and from 12 to 28%, respectively). Decrease in cell-surface hydrophobicity was also observed by salt aggregation test. The results obtained can be used to determine the therapeutic concentrations of antibiotics to induce changes in the adhesion properties of bacteria.
Insights
Antibiotic treatment alters Pseudomonas aeruginosa outer membrane proteins, affecting bacterial adhesion in orthopaedic wound infections. This research helps determine antibiotic concentrations to modify bacterial surface properties.
Area of Science:
- Microbiology
- Biochemistry
- Orthopaedic Surgery
Background:
- Nosocomial wound infections caused by Pseudomonas aeruginosa are a growing concern in orthopaedic surgery.
- Bacterial outer membrane protein composition and cell-surface hydrophobicity significantly influence adhesion to tissues and medical devices.
Purpose of the Study:
- To examine the outer membrane protein (OMP) composition of Pseudomonas aeruginosa strains isolated from orthopaedic wound infections.
- To investigate the impact of common antibiotics (cefotaxime, amoxicillin/clavulanic acid, amikacin) on OMP profiles and cell-surface hydrophobicity.
Main Methods:
- Capillary electrophoresis was used to analyze the OMP patterns of six Pseudomonas aeruginosa strains.
- The salt aggregation test was employed to assess changes in cell-surface hydrophobicity after antibiotic treatment.
Main Results:
- Each bacterial strain exhibited a unique OMP electrophoretic pattern.
- Antibiotic treatment significantly altered OMP composition, with notable changes observed in strains KT 2, KT 7, and KT 28.
- Amikacin decreased a specific 38,800 Da protein in strain KT 2, while cefotaxime increased it in strains KT 7 and KT 28.
- A decrease in cell-surface hydrophobicity was observed post-antibiotic treatment.
Conclusions:
- Antibiotic administration can significantly modify the outer membrane protein profile and adhesion properties of Pseudomonas aeruginosa.
- Findings can inform the selection of therapeutic antibiotic concentrations to manage bacterial adhesion in orthopaedic wound infections.

