Simple technique for the assay of antibiotic synergism against enterococci

A simple technique with antibiotic-impregnated disks has been developed to demonstrate the antibiotic synergism of penicillins and aminoglycosides against enterococci. The two antibiotic disks are placed on inoculated plates so that the distance between their centers is less than the sum of the two radii of their previously determined inhibitory zones. After 5 h of incubation, beta-lactamase powder is dusted onto the susceptibility plate with a sterile cotton swab. After overnight incubation, the synergistic effect of the antibiotics is demonstrated by a clear area of no growth in between the two disks. There is good correlation between the synergism determined by this procedure and the susceptibility of the organisms to 2 mg of streptomycin per ml. This technique is simple and can be employed by any laboratory which performs Kirby-Bauer susceptibility procedures.