Bacteriostatic and Bactericidal Activities of Selected Beta-Lactam Antibiotics Studied on Agar Plates

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Abstract
A novel and time-saving method for assessing bactericidal activities of β-lactam antibiotics on agar plates is described. Minimal inhibitory concentrations (MICs) were determined by the agar dilution method. A potent β-lactamase solution was sprayed onto the plates to inactivate the antibiotic. After further incubation at 37°C overnight, the minimal concentration at which no visible growth occurred on the plates was defined as minimal bactericidal concentration (MBC). With undiluted culture as the inoculum, strains of Staphylococcus aureus, Escherichia coli , and Klebsiella pneumoniae showed a marked increase in MBC values compared with the values of MIC. There was a marked decrease in both the MICs and MBCs with diminution of inoculum size. The two concentrations were almost the same when the inoculum size was decreased to a 10 −4 dilution. In contrast, MIC and MBC for enterococci showed no marked decrease with decrease in inoculum size. Although the present study was performed with β-lactamase-unstable penicillins and cephalosporins, the method can be applicable with any β-lactam antibiotic if optimal and potent enzymes are available.