Yung-Yuan Kwok

Susceptibility of Bacteroides fragilis to six antibiotics determined by standardized antimicrobial disc susceptibility testing.

The susceptibility of 100 or more strains of Bacteroides fragilis to six antibiotics was determined by standardized agar dilution and disc diffusion tests. Good correlation of results of the two methods was obtained with chloramphenicol, clindamycin, lincomycin, penicillin, and vancomycin. Correlation of results with erythromycin was not as good. A great deal of overlapping of zone diameters among strains classed as susceptible, intermediate, and resistant occurred with both erythromycin and lincomycin, making interpretation of disc diffusion tests difficult. All strains tested were susceptible to chloramphenicol; 94% were susceptible to clindamycin at concentrations readily achieved with ordinary dosage, and all strains were inhibited by 6.2 μg/ml, a level which is achieved with somewhat more intensive therapy. Only 7% were shown to be susceptible to erythromycin and 13% to lincomycin under the conditions of our testing procedure. Only a small percentage of the strains were susceptible to penicillin, and none was susceptible to vancomycin.

Standardized antimicrobial disc susceptibility testing of anaerobic bacteria: in vitro susceptibility of Clostridium perfringens to nine antibiotics.

The in vitro susceptibility of 43 strains of Clostridium perfringens to nine antibiotics was determined by a standardized test for rapid-growing anaerobes. Good correlation was established between the agar dilution susceptibility and the disc diffusion susceptibility results. The inhibition zone diameters around the antibiotic discs, however, were generally much smaller than those of gram-negative anaerobes previously studied. All of the strains tested were susceptible in vitro to chloramphenicol, clindamycin, doxycycline, minocycline, penicillin, and vancomycin. Erythromycin showed poor in vitro activity against this organism, with only 7% of the strains susceptible, 72% intermediate in susceptibility, and 21% resistant. In tests of the 43 strains against lincomycin, 58% were susceptible, 32.5% were intermediate in susceptibility, and 9.5% were resistant. Against tetracycline, 37% of the strains were intermediate in susceptibility and the rest were susceptible.

Susceptibility of Clostridium ramosum to Antimicrobial Agents

The susceptibility of 49 strains of Clostridium ramosum to 10 antibiotics was determined by agar dilution and disk diffusion tests. Results showed that, among the anaerobes, C. ramosum is second only to Bacteroides fragilis in its resistance to antimicrobial agents. All strains were susceptible to penicillin, carbenicillin, chloramphenicol, vancomycin, and metronidazole at readily achievable blood levels. Most strains (83%) were susceptible to erythromycin. There was a high level of resistance to clindamycin in 16% of the strains. All isolates were resistant to rifampin and gentamicin, and most were resistant to lincomycin. Assessment of susceptibility by measurement of inhibition zone diameters with disk diffusion tests was not satisfactory.

Disk Susceptibility Testing of Slow-Growing Anaerobic Bacteria

The susceptibility of 55 strains of slow-growing anaerobes to eight clinically useful or potentially useful antibiotics was determined by agar dilution and disk diffusion tests. Strains of the genera Peptococcus, Peptostreptococcus, Megasphaera, Veillonella, Eubacterium, Bifidobacterium, Clostridium, and Fusobacterium were included. All strains were susceptible to chloramphenicol, but varied in their susceptibility to penicillin, lincomycin, clindamycin, tetracyclines, and vancomycin. Correlation between minimal inhibitory concentration and inhibition zone diameters was generally good. Prediction of susceptibility based on zone diameter measurements appeared satisfactory. Although routine susceptibility testing of anaerobic bacteria is not recommended, there are circumstances where such testing is relevant to the clinical situation. For those laboratories ill-equipped to do dilution tests, a disk diffusion test would give relatively accurate preliminary information. Quantitative susceptibility tests could then be done by a reference laboratory.

Effect of carbon dioxide on in vitro susceptibility of anaerobic bacteria to erythromycin.

The activity of erythromycin against 317 strains of anaerobic bacteria, including 133 strains of the Bacteroides fragilis group, was tested by the agar dilution method in an anaerobic atmosphere with two different concentrations of carbon dioxide and without CO2. The effect of the atmosphere of incubation on the agar surface pH was also determined. All strains grew well in the GasPak (GP) environment. However, 3.5 and 30.3% of strains failed to grow in the 2 and 0% CO2 environments, respectively. The quality of growth was best in the GP environment and poorest in the 0% CO2 environment. Minimal inhibitory concentrations in the GP and 2% CO2 environments were frequently the same or one dilution lower in the 0% than in the GP environment. In the 0% CO2 atmosphere, minimal inhibitory concentrations were usually two to three dilutions lower than in the GP environment. Consequently, only 24% of B. fragilis strains were susceptible to erythromycin in the GP environment, whereas 77% were susceptible in the 0% CO2 environment. For Fusobacterium species, 12% were susceptible to erythromycin in the GP environment, and 73% were susceptible in the 0% CO2 environment. There was a comparable decrease in pH in all three atmospheres tested. In vitro susceptibility testing of erythromycin against anaerobic bacteria should be performed in an atmosphere containing carbon dioxide.

Susceptibility of Gardnerella vaginalis to cephradine.

The activity of cephradine and the influence of pH on its activity against 70 Gardnerella vaginalis strains were determined. Serial dilutions of cephradine (0.062 to 256 micrograms/ml) were incorporated into Dunkelberg agar, inoculated with a Steers replicator, incubated in 5% CO2 for 48 h, and examined. The minimal inhibitory concentrations for 90% of the isolates were 16 and 8.0 micrograms/ml at pH 7.0 to 6.0 and 5.5, respectively.

Absence of tolerance to cefoxitin in anaerobic bacteria.

None of the 46 strains of anaerobic bacteria tested, including 26 strains of Bacteroides fragilis, showed tolerance (minimal bactericidal concentration/minimum inhibitory concentration ratio >32) to cefoxitin.