Antimicrobial Susceptibility Testing of Bacillus anthracis : Comparison of Results Obtained by Using the National Committee for Clinical Laboratory Standards Broth Microdilution Reference and Etest Agar Gradient Diffusion Methods

Abstract

ABSTRACT We determined the patterns of antimicrobial susceptibility of 65 isolates of Bacillus anthracis (50 historical and 15 recent U.S. clinical isolates) to nine antimicrobial agents using the National Committee for Clinical Laboratory Standards (NCCLS) broth microdilution reference method. The results for the 50 historical B. anthracis isolates obtained by the broth microdilution method were compared to those generated by the Etest agar gradient diffusion method. One isolate of B. anthracis was β-lactamase positive and resistant to penicillin (MIC, 128 μg/ml); a second isolate, which was β-lactamase negative, was borderline penicillin resistant, with the penicillin MICs for the isolate varying from 0.12 to 0.25 μg/ml; and the remainder of the isolates were β-lactamase negative and penicillin susceptible (MICs, ≤0.12 μg/ml). Approximately 78% of the isolates showed reduced susceptibility to ceftriaxone (MICs, ≥16 μg/ml). All B. anthracis isolates were susceptible to chloramphenicol (MICs, ≤8 μg/ml), ciprofloxacin (MICs, ≤ 1 μg/ml), clindamycin (MICs, ≤0.5 μg/ml), rifampin (MICs, ≤0.5 μg/ml), tetracycline (MICs, ≤0.06 μg/ml), and vancomycin (MICs, ≤2 μg/ml) by use of NCCLS breakpoints for staphylococci. All 15 recent B. anthracis isolates from the United States were susceptible to penicillin, doxycycline, and ciprofloxacin. By use of the susceptibility breakpoint for staphylococci of ≤0.5 μg/ml, 97% of the B. anthracis isolates tested would have been categorized as intermediate to erythromycin. No statistically significant difference was found between the results of broth microdilution testing and the results of the Etest method for any of the antimicrobial agents tested; however, the results for penicillin obtained by the Etest were 1 to 9 dilutions lower than those obtained by the broth microdilution method. The differences in the penicillin MICs by the Etest method and the difficulties of reading the Etest results through the glass of a biological safety cabinet may limit the utility of this alternate susceptibility testing method for B. anthracis isolates.