Comparative antibacterial activities of 7 alpha-methoxy cephalosporins and 7 beta-methoxyiminoacetamido cephalosporins against Bacteroides fragilis

The in vitro antibacterial activities of the newly developed 7 alpha-methoxy cephalosporins and 7 beta-methoxyiminoacetamido cephalosporins against 67 clinical isolates of Bacteroides fragilis and their resistance to the hydrolytic action of a beta-lactamase produced by B. fragilis were simultaneously compared. The minimal inhibitory concentrations that inhibited 90% of the 7 alpha-methoxy cephalosporins, cefoxitin, cefmetazole, moxalactam, and cefotetan, against the isolates were 4, 8, 8, and 16 micrograms/ml, respectively, and these antibiotics were entirely resistant to hydrolysis by beta-lactamases (0.10 mumol/h per mg of protein) of the isolates. By contrast, 7 beta-methoxyiminoacetamido cephalosporins represented by cefotaxime, ceftizoxime, and cefmenoxime were not effective, as indicated by the minimal inhibitory concentrations that inhibited 90%, 64, 32, and 128 micrograms/ml, respectively. Their antibacterial activities clearly corresponded to their resistance to the hydrolytic action of the beta-lactamase: namely, the correlation coefficients in regression curves of cefotaxime, ceftizoxime, and cefmenoxime, which were expressed by the antibacterial activity (x axis) and the beta-lactamase activity (y axis) were 0.098, 0.034, and 0.163, respectively.     

Comparison of the inoculum effect of cefoxitin and other cephalosporins and of beta-lactamase inhibitors and their penicillin-derived components on the Bacteroides fragilis group.

We compared the inoculum effects for 109 recent clinical isolates of the Bacteroides fragilis group of cefoxitin, cefotetan, ceftizoxime, ceftriaxone, and three beta-lactamase inhibitors (clavulanic acid, sulbactam, and tazobactam) and their penicillin-derived components. Bactericidal activity was assayed and morphologic changes were monitored for selected strains exhibiting a large inoculum effect. Ceftizoxime demonstrated the largest inoculum effect, followed by cefotetan and ceftriaxone. The large inoculum effect of ceftizoxime and ceftriaxone was correlated with filamentous transformation at the high inoculum (10(8) CFU/ml) and lack of bactericidal activity suggesting drug destruction or inactivation. Cefotetan was bactericidal for B. fragilis isolates but not for other members of the B. fragilis group. Cefoxitin showed the least inoculum effect and was consistently bactericidal at high (10(8) CFU/ml), standard (10(6) CFU/ml), and low (10(4) CFU/ml) inocula, followed by ampicillin-sulbactam. Piperacillin-tazobactam and ticarcillin-clavulanic acid showed an intermediate inoculum effect. The degree of inoculum effect observed generally correlated with bactericidal activity at all inocula.     

Six-year retrospective survey of the resistance of Bacteroides fragilis group species to clindamycin and cefoxitin

Two hundred and forty-six strains of the Bacteroides fragilis group, all clinical isolates, collected at the Wadsworth Veterans Administration Medical Center from 1977 to 1982, were tested for susceptibility to clindamycin and cefoxitin. There was no significant change in resistance to either clindamycin or cefoxitin over the time period tested for any individual species, nor for the B. fragilis group in toto. Striking differences in susceptibility to the two drugs were seen among species of the B. fragilis group. B. fragilis displayed resistance to cefoxitin (32 micrograms/ml) and clindamycin (8 micrograms/ml) of 0.0% and 0.8%, respectively, whereas B. thetaiotaomicron showed resistances of 12.7% to cefoxitin (32 micrograms/ml) and 9% to clindamycin (8 micrograms/ml). B. thetaiotaomicron, B. distasonis, and B. ovatus are distinctly more resistant to cefoxitin than B. fragilis and B. vulgatus. Similarly B. thetaiotaomicron and B. distasonis are much more resistant to clindamycin than are the other B. fragilis group species. It is apparent that determination of species within the B. fragilis group is important in evaluating a potential therapeutic regimen.     

Cefoxitin inactivation by Bacteroides fragilis.

We have surveyed the susceptibility of 1,575 clinical isolates of the Bacteroides fragilis group of organisms to cefoxitin and eight other antimicrobial agents. Eleven isolates, 0.7% of the total, were highly cefoxitin resistant and had minimum inhibitory concentrations of greater than or equal to 64 micrograms/ml. These isolates were also resistant to other beta-lactam antibiotics. Of 11 isolates, 4 were able to inactivate cefoxitin in broth cultures, as measured by microbiological and high-pressure liquid chromatography assays. Two distinct patterns of cefoxitin breakdown products were detected by high-pressure liquid chromatography analysis. The beta-lactamase inhibitors clavulanic acid and sulbactam failed to show synergism with cefoxitin. These data demonstrate that members of the B. fragilis group have acquired a novel resistance mechanism enabling them to inactivate cefoxitin.     

Effect of beta-lactamase inhibitors on the antimicrobial activity of cefoperazone, cefotaxime, and ceftizoxime against aerobic and anaerobic beta-lactamase producing bacteria

Clavulanic acid (2.0 micrograms/ml) lowered the minimal inhibitory concentrations (MICs) of ceftizoxime and cefotaxime against 49 strains of the Bacteroides fragilis group by a mean of 4.0 and 3.4 log2 concentrations, respectively. Sulbactam plus ceftizoxime gave almost identical results. Sulbactam lowered cefoperazone MICs by a mean of 2 log2 concentrations. Against 52 aerobic and facultative isolates producing a variety of beta-lactamase types, the use of beta-lactamase inhibitors (sulbactam) was most effective in reducing the MICs of cefoperazone.     

In-vitro study of the susceptibility of cefoxitin/cefotetan resistant Bacteroides fragilis group strains to various other antimicrobial agents

The in-vitro activity of various beta-lactam antibiotics, beta-lactam/beta-lactamase inhibitor combinations, clindamycin, and metronidazole was determined against Bacteroides fragilis group isolates that were resistant to both cefoxitin and cefotetan. Among the cephalosporins tested ceftizoxime was the most active with 80% of the strains susceptible, followed by cefotaxime (65% susceptible), and cefoperazone (47% susceptible). Piperacillin and clindamycin showed comparable activity to ceftizoxime with 80% and 81% susceptible, respectively. The addition of a beta-lactamase inhibitor (sulbactam, clavulanate, or tazobactam) enhanced the activity of the various beta-lactam agents from 4-fold to 16-fold overall. One strain of B. fragilis was found that was resistant to all beta-lactam agents either alone or in combination. All strains in this study were susceptible to less than or equal to 4 mg/l of metronidazole.     

Comparison of the bactericidal activity of clindamycin and metronidazole against cefoxitin-susceptible and cefoxitin-resistant isolates of the Bacteroides fragilis group.

Time-kill kinetic methodology was used to evaluate the bactericidal activity of cefoxitin, cefotetan, clindamycin, and metronidizole against cefoxitin-susceptible and cefoxitin-resistant isolates of the Bacteroides fragilis group. Overall, metronidazole was the most bactericidal agent, with all isolates being killed with less than or equal to 4 micrograms/ml at 24 hr. Clindamycin was the next most bactericidal agent, with 20 of 26 isolates being killed with less than 16 micrograms/ml. Six isolates with clindamycin MICs greater than or equal to 64 micrograms/ml were not killed at 24 hr, with concentrations as high as 256 micrograms/ml. Cefoxitin and cefotetan were the least bactericidal agents tested. Seven isolates with MICs of greater than or equal to 64 micrograms/ml to each agent demonstrated a lack of killing at 24 hr, with concentrations of the respective agent as high as 256 micrograms/ml. At concentrations with either agent of 32 micrograms/ml, the remaining 19 isolates were killed at 24 hr. Of the six B. fragilis isolates resistant to clindamycin, four were also resistant to both cefoxitin and cefotetan. We conclude that in hospitals with cefoxitin-resistant B. fragilis group isolates, metronidazole would provide appropriate therapy.     

Multicenter Study of In Vitro Susceptibility of the Bacteroides fragilis Group, 1995 to 1996, with Comparison of Resistance Trends from 1990 to 1996

Antimicrobial resistance, including plasmid-mediated resistance, among the species of the Bacteroides fragilis group is well documented. An analysis of the in vitro susceptibility of B. fragilis group species referred between 1995 and 1996 as well as during a 7-year (1990 to 1996), prospective, multicenter survey of over 4,000 clinical isolates of B. fragilis group species was undertaken to review trends in the percent resistance to and geometric mean MICs of the antibiotics tested. There was a trend toward a decrease in the geometric mean MICs of most beta-lactam antibiotics, while the percent resistance to most agents was less affected. Within the species B. fragilis, the geometric mean MICs showed significant (P < 0.05) decreases for piperacillin-tazobactam, ticarcillin-clavulanate, piperacillin, ticarcillin, ceftizoxime, cefotetan, and cefmetazole; a significant increase was observed for clindamycin and cefoxitin. For the non-B. fragilis species, a significant decrease in the geometric mean MICs was observed for meropenem, ampicillin-sulbactam, ticarcillin-clavulanate, piperacillin, ticarcillin, ceftizoxime, and cefmetazole; a significant increase was observed for cefoxitin. Significant increases in percent resistance were observed within the B. fragilis strains for ticarcillin and ceftizoxime and within the non-B. fragilis isolates for cefotetan. Significant increases in percent resistance among all B. fragilis group species were observed for clindamycin, while imipenem showed no significant change in resistance trends. The trend analysis for trovafloxacin was limited to 3 years, since the quinolone was tested only in 1994, 1995, and 1996. During the 7 years analyzed, there was no resistance to metronidazole or chloramphenicol observed. The data demonstrate that resistance among the B. fragilis group species has decreased in the past several years, the major exception being clindamycin. The majority of the resistance decrease has been for the beta-lactams in B. fragilis, compared to other species. The reasons for these changes are not readily apparent.     

Comparative in vitro activities of a new quinolone, WIN 57273, and piperacillin plus tazobactam against anaerobic bacteria.

The in vitro activities of a new quinolone, WIN 57273, and the combination of piperacillin and tazobactam, a beta-lactamase inhibitor, were compared with those of cefoxitin, ceftizoxime, chloramphenicol, clindamycin, imipenem, metronidazole, and piperacillin for 123 clinical anaerobic isolates. Ceftizoxime and cefoxitin had equivalent activities, while metronidazole was active against gram-negative isolates. In the Bacteroides fragilis group, species other than B. fragilis were the most resistant. The combination of piperacillin with tazobactam in a ratio of 8 to 1 was more effective than piperacillin against B. fragilis group organisms when the MIC of piperacillin was greater than or equal to 64 micrograms/ml. Overall, WIN 57273 (i) and imipenem (ii) were the most active agents, with MICs for 50 and 90% of strains of (i) 0.25 and 0.5 and (ii) 0.125 and 2 microgram/ml, respectively.     

Comparative antimicrobial activity of aminothiazolyl methoxyimino cephalosporins against anaerobic bacteria, including 100 cefoxitin-resistant isolates

Three aminothiazolyl methoxyimino cephalosporins (ceftizoxime, ceftriaxone, and cefotaxime with and without its metabolite) were tested against 500 strains of anaerobic bacteria using the NCCLS reference agar dilution procedure. Eighty-seven percent of all strains tested were from the Bacteroides fragilis group. When tested against a collection of 100 cefoxitin-resistant isolates, ceftizoxime and the cefotaxime/desacetyl-cefotaxime combination were the most active in vitro, inhibiting 32-38% of strains. Ceftriaxone inhibited the greatest number (87%) of cefoxitin-susceptible anaerobes at less than or equal to 32 micrograms/ml. A regional variation in the activity of these drugs was confirmed when an additional 300 isolates were examined from three medical centers. Cefotaxime, ceftizoxime, and ceftriaxone were essentially equal in overall antimicrobial activity, although each drug was judged the best at one of the three locations. Cefoxitin resistance (MIC greater than or equal to 32 micrograms/ml) was consistent among the institutions at a 25% incidence for all organisms tested, and 28% cefoxitin resistance among the B. fragilis group strains. Cefoxitin resistance was not determined to be associated with a beta-lactamase mechanism, but ceftriaxone and other aminothiazolyl cephems were hydrolyzed, thus elevating their MICs. Regional variation in anaerobic organism susceptibility to cephamycins and cephalosporins in company with the variable beta-lactam resistance mechanisms seems to require periodic, epidemiologic monitoring of in vitro drug activity by appropriate methods to assure continued antimicrobial efficacy.     

Survey of Bacteroides fragilis group susceptibility patterns in Canada.

The in vitro activities of penicillin, clindamycin, chloramphenicol, metronidazole, piperacillin, piperacillin-tazobactam, ticarcillin, ticarcillin-clavulanate, ampicillin-sulbactam, cefoxitin, ceftizoxime, cefotetan, moxalactam, and imipenem against 348 Bacteroides fragilis group isolates collected from six Canadian cities during 1990 were determined by the National Committee for Clinical Laboratory Standards (NCCLS) agar dilution technique. All isolates were susceptible to chloramphenicol, metronidazole, piperacillin-tazobactam, and imipenem. For the other antibiotics tested, the following resistance rates were observed: penicillin, 97%; clindamycin, 9%; piperacillin, 19%; ticarcillin, 31%; ticarcillin-clavulanate, 0.28%; ampicillin-sulbactam, 0.85%; cefoxitin, 26%; ceftizoxime, 15%; cefotetan, 53%; and moxalactam, 17%. Susceptibility profiles to beta-lactam antibiotics varied among the different species tested: B. fragilis and Bacteroides vulgatus demonstrated lower resistance rates than Bacteroides distasonis and indole-positive Bacteroides thetaiotaomicron and Bacteroides ovatus. Ceftizoxime results should be interpreted cautiously, because the MICs obtained with the recommended NCCLS control strain were lower than expected.     

Differences in the in vitro inhibitory and bactericidal activity of ceftizoxime, cefoxitin, cefotetan, and penicillin G against Bacteroides fragilis group isolates. Comparison of time-kill kinetic studies with MIC values

Nineteen strains of the Bacteroides fragilis group were used to determine the bactericidal activity of ceftizoxime, cefoxitin, cefotetan, and penicillin G with time-kill kinetics studies. Each antimicrobial agent was tested at subinhibitory (1/2 X MIC), inhibitory (1 X MIC), and suprainhibitory (4 X MIC) concentrations. Penicillin G exhibited virtually no sustained bactericidal activity at any of the antimicrobial concentrations tested. At subinhibitory concentrations, ceftizoxime was considerably more bactericidal than cefoxitin or cefotetan: At 12 hr, ceftizoxime killed 89% of the inoculum, whereas cefoxitin and cefotetan killed 35% and 33% of the inoculum, respectively. At inhibitory concentrations, ceftizoxime was again more bactericidal than cefoxitin and cefotetan: At 12 hr, ceftizoxime killed 90% of the inoculum, whereas cefoxitin and cefotetan killed 78% and 73%, respectively. At suprainhibitory concentrations, all three antimicrobial agents showed comparable bactericidal activity at 12 and 24 hr. Ceftizoxime and cefoxitin had somewhat lower killing rates overall against test strains with high MICs (greater than or equal to 32) versus low MICs (less than or equal to 16). However, at subinhibitory concentrations, ceftizoxime killed the B. fragilis group strains with high or low MIC values more effectively than cefotetan killed strains with low MICs. At the highest antibiotic concentrations tested (4 X MIC), only slight differences were seen in the bactericidal activity of the three compounds, regardless of MICs.     

Inhibitory and bactericidal activity of selected beta-lactam agents alone and in combination with beta-lactamase inhibitors compared with that of cefoxitin and metronidazole against cefoxitin-susceptible and cefoxitin-resistant isolates of the Bacteroides fragilis group.

The inhibitory activity of five beta-lactam agents, alone and in combination with a beta-lactamase inhibitor, was compared with that of cefoxitin and metronidazole against 300 beta-lactamase producing Bacteroides fragilis group isolates. Each of the beta-lactamase inhibitors significantly potentiated the activity of the respective beta-lactam. In the presence of clavulanate, the MIC90 (minimum inhibitory concentration) values of amoxicillin and ticarcillin were reduced 64-fold and 32-fold, respectively. Similarly, sulbactam enhanced the activity of ampicillin and cefoperazone 16-fold and 8-fold, respectively, whereas tazobactam potentiated the activity of piperacillin 16-fold. Few strains were resistant to the beta-lactam-beta-lactamase inhibitor combinations and were comprised of strains of B. fragilis, B. thetaiotamicron, and B. distasonis. Of the strains, 7% were resistant to cefoxitin, and none to metronidazole. Using time-kill kinetic studies, the bactericidal activity of the various beta-lactam agents, with and without beta-lactamase inhibitors, was determined and compared with that of cefoxitin and metronidazole against cefoxitin-susceptible and cefoxitin-resistant isolates of the B. fragilis group. Overall, metronidazole was the most bactericidal agent with all isolates being killed with less than or equal to 4 micrograms/ml at 24 hr. Ampicillin-sulbactam was the next most bactericidal agent with all isolates being killed with less than or equal to 16/8 micrograms/ml of ampicillin-sulbactam at 24 hr. Amoxicillin-clavulanate and cefoperazone-sulbactam had bactericidal activity similar to that of ampicillin-sulbactam. Piperacillin-tazobactam and ticarcillin-clavulanate were bactericidal at higher concentrations with all isolates killed with 64 micrograms/ml of piperacillin and 128 micrograms/ml of ticarcillin combined with their respective beta-lactamase inhibitors. None of the beta-lactam agents alone was able to kill more than 19 of the 26 isolates. We conclude that beta-lactam agents combined with beta-lactamase inhibitors have both inhibitory and bactericidal activity against cefoxitin-resistant members of the B. fragilis group provided that the concentrations achieved for these combinations are at the upper limits for maximum recommended dosing. Although isolates of the B. fragilis group have been reported to produce unusual beta-lactamases that are refractory to beta-lactamase inhibitors, none of the cefoxitin-resistant isolates tested in this study were resistant to the beta-lactam-beta-lactamase inhibitor combinations.     

Role of Bacteroides bivius beta-lactamase in beta-lactam susceptibility

The susceptibility of 46 clinical isolates of Bacteroides bivius to amoxicillin, cefotaxime, cefoxitin, ceftizoxime, cephaloridine, cephalothin, moxalactam, penicillin G, amoxicillin plus clavulanic acid in a ratio of 2:1, carbenicillin, cefamandole, and ceftazidime was determined by an agar dilution technique. For the first eight agents susceptibility testing was also done with the addition of clavulanic acid (0.75 microgram/ml). For all agents, beta-lactamase-positive strains (35, using a nitrocefin slide test) were inhibited at higher concentrations than beta-lactamase-negative strains. Clavulanic acid reduced the susceptibility of the beta-lactamase-positive strains to the level of the beta-lactamase-negative strains to all agents. We prepared crude extracts of beta-lactamase from six strains. Activity against nitrocefin was directly related to their susceptibilities. The beta-lactamase had a mixed-substrate profile, hydrolyzing both penicillins and cephalosporins. Our results suggest a slow inactivation of cefoxitin, ceftizoxime, and moxalactam by the beta-lactamase. Clavulanic acid and cefoxitin inhibited the enzyme, whereas p-hydroxymercuribenzoate and cloxacillin did not. Thus, there was a clear relationship between beta-lactamase activity and susceptibility to beta-lactams, including cefoxitin and third-generation cephalosporins. The substrate and inhibition profiles of the B. bivius beta-lactamase were different from those of enzymes found in the "B. fragilis group."     

The antimicrobial susceptibility patterns of the Bacteroides fragilis group in the United States, 1987

A nationwide survey to monitor the susceptibility of the Bacteroides fragilis group, which began in 1981, was continued during 1987. In addition to the eleven drugs evaluated in 1986, sulbactam, a potent beta-lactamase inhibitor, was tested alone and in combination with ampicillin and cefoperazone. Imipenem, ampicillin/sulbactam, cefoperazone/sulbactam, and ticarcillin/clavulanic acid were the most active newer drugs tested, with less than 1% resistance rates. Chloramphenicol, metronidazole and clindamycin also had excellent activity with resistance rates of 0%, 0%, and 3% respectively. Resistance rates to cefoxitin remained stable at 8%. Ceftizoxime and cefotetan had resistance rates of 26% and 29%, respectively. Rates of resistance varied among different institutions and between the various species.     

In vitro activity of cefbuperazone compared with that of other new beta-lactam agents against anaerobic gram-negative bacilli and contribution of beta-lactamase to resistance.

Cefbuperazone was compared with other currently available and investigational antibiotics against 278 clinical isolates of anaerobic gram-negative bacilli by an agar dilution method. Cefbuperazone and cefotetan were equally active against Bacteroides fragilis, with 8% of the organisms tested found to be resistant to 32 micrograms of either drug per ml. Cefoperazone, cefotaxime, ceftriaxone, and cefmetazole were less active against these strains; cefoxitin, moxalactam, piperacillin, clindamycin, and metronidazole were more active. None of the agents were consistently active against any of the other anaerobic gram-negative bacilli except imipenem, for which the minimum concentration required to inhibit 90% of all strains tested was 4 micrograms/ml. A 10,000-fold increase in inoculum size caused an increase in the MIC of ceftriaxone, cefotaxime, and cefoperazone but not of cefbuperazone, cefotetan, or cefoxitin. Investigation of the mechanism of resistance to cephalosporin-like agents demonstrated a correlation between the level of resistance and beta-lactamase activity. Cefbuperazone, cefotetan, and cefoxitin were not hydrolyzed, had lower MICs, and were less affected by changes in inoculum size than were cefotaxime, ceftriaxone, and cefoperazone.     

Inactivation of cefoxitin and moxalactam by Bacteroides bivius beta-lactamase.

Moxalactam and cefoxitin are known for their high stability against Bacteroides beta-lactamases. We investigated the beta-lactamase activity of crude extracts obtained from three strains of Bacteroides bivius and two strains of Bacteroides fragilis against cefoxitin and moxalactam. In a spectrophotometric antibiotic assay with a 24-h incubation period, B. bivius extracts decreased the initial concentration (10 micrograms/ml) of moxalactam and cefoxitin by 60%, whereas B. fragilis extracts had no effect. In a microbiological assay, when B. bivius or B. fragilis extracts were added to cephalothin (10 micrograms/ml) or cefamandole (4 micrograms/ml), we observed complete disappearance of the inhibitory zones against the indicator strain (Clostridium perfringens ATCC 13124). Only the B. bivius extracts were able to decrease the inhibitory activity (from 10 to 100%) of cefoxitin and moxalactam (each at 10 micrograms/ml). Prior addition of clavulanic acid to crude extracts prevented the losses of antibacterial activity. Furthermore, the inhibition of the beta-lactamase hydrolysis of nitrocefin by cefoxitin or moxalactam was prevented by a 12-h preincubation of the beta-lactam with the B. bivius extracts but not with the B. fragilis extracts. Finally, with the B. bivius strain producing the most beta-lactamase, we showed an effect of inoculum size on the MICs of cefoperazone, cefoxitin, and moxalactam with a broth dilution technique. Increasing the inoculum size with the B. fragilis strains had no effect on the MISs of cefoxitin and moxalactam. These results indicate a slow and clavulanate-sensitive beta-lactamase activity of B. bivius extracts against cefoxitin and moxalactam.     

In-vitro activity of amoxycillin and ticarcillin in combination with clavulanic acid compared with that of new beta-lactam agents against species of the Bacteroides fragilis group

One hundred and one isolates representing five species of the Bacteroides fragilis group were examined for their susceptibility to amoxycillin and ticarcillin alone or in combination with clavulanic acid, as well as to cefoxitin, latamoxef (moxalactam), ceftizoxime, imipenem, chloramphenicol, clindamycin and metronidazole by the proposed NCCLS agar dilution method. Bactericidal activity of amoxycillin, ticarcillin, combinations of amoxycillin-clavulanic acid and ticarcillin-clavulanic acid, cefoxitin and latamoxef against 54 strains was further assessed by a dilution test in broth. The most active agents were imipenem and the combinations of amoxycillin-clavulanic acid and ticarcillin-clavulanic acid. MIC results indicated that Bacteroides fragilis and B. vulgatus were more susceptible to beta-lactams than were B. distasonis, B. ovatus and B. thetaiotaomicron. Clavulanic acid greatly potentiated the activity of amoxycillin and ticarcillin against the 94 beta-lactamase positive strains but had no effect on the seven B. distasonis isolates that were beta-lactamase negative. Synergistic bactericidal activities could be observed by the amoxycillin-clavulanic and ticarcillin-clavulanic combinations.     

In vitro activities of three of the newer quinolones against anaerobic bacteria.

The antimicrobial activities of three new quinolone compounds, sparfloxacin, temafloxacin, and WIN 57273, against anaerobic bacteria were determined in three separate studies. The Wadsworth agar dilution technique using brucella-laked blood agar was used throughout. The activities of other antimicrobial agents, including ciprofloxacin, imipenem, chloramphenicol, metronidazole, cefotetan, cefoxitin, and amoxicillin-clavulanic acid, were also determined. The breakpoints of the new quinolones were 2 micrograms/ml for sparfloxacin and WIN 57273 and 4 micrograms/ml for temafloxacin. WIN 57273 displayed very good activity against anaerobes, inhibiting all strains of Bacteroides fragilis group species at 2 micrograms/ml. Only two strains of Fusobacterium species were resistant (MIC, 4 micrograms/ml). Sparfloxacin inhibited 78% of B. fragilis strains and 44% of other B. fragilis group isolates at 2 micrograms/ml. At 2 micrograms/ml, the percentages of other anaerobic species susceptible were as follows: B. gracilis, 70%; other Bacteroides species, 61%; Clostridium species, 50%; Fusobacterium species, 70%; Peptostreptococcus species, 91%; non-spore-forming gram-positive rods, 71%. Temafloxacin inhibited 91% of B. fragilis strains and 87% of other B. fragilis group species at 4 micrograms/ml. All strains of other Bacteroides species, 78% of Fusobacterium species, 80% of Clostridium species, and 90% of Peptostreptococcus species were inhibited at 4 micrograms of temafloxacin per ml.     

Comparative activity of several beta-lactam antibiotics against anaerobes determined by two methods

The susceptibility of 120 strains of several species of anaerobes to a number of second and third generation beta-lactam antibiotics was determined by the National Committee for Clinical Laboratory Standards reference agar dilution and microdilution methods. The antibiotics tested were cefoperazone, cefotaxime, cefotetan, ceftizoxime, cefoxitin, and imipenem. The MIC50s ranged from 0.125 to 16 micrograms/ml. The MIC90s were lowest with imipenem at 0.5 micrograms/ml, followed by cefoxitin at 32 micrograms/ml; they were highest with cefotetan at 128 micrograms/ml and were 64 micrograms/ml with the others. In vitro drug activity varied with the antibiotic, the organism, the method used, and the breakpoint selected. Rates of resistance varied considerably between the taxonomic groups of organisms tested and also among species within a group. Overall, reproducibility with the agar dilution method ranged from 44% to 85%; testing with ceftizoxime was the least reproducible. Microdilution results agreed within +/- 1 dilution of the agar dilution mode 79% to 95% of the time, with some variation between drugs and organisms tested. Because there were distinct differences in the activity of some drugs against certain species, no antibiotic can substitute for others in in vitro testing.