Major methodology-dependent discordant susceptibility results for Bacteroides fragilis group isolates but not other anaerobes.

Two standardized susceptibility test methods, a broth microdilution (BMD) and agar dilution (AD) method were performed on a total of 441 clinical isolates of anaerobes with ceftizoxime, cefotaxime, ceftriaxone, cefoxitin, piperacillin, and metronidazole. Against the 339 strains of the Bacteroides fragilis group BMD minimum inhibitory concentration (MIC) values were lower than those from AD testing for all the beta-lactams. Overall for the B. fragilis group and the beta-lactams, the mode MIC values were two- to 64-fold lower, and the MIC50 values two- to eightfold lower. Resistance rates were 11%-28% higher overall with AD results and were higher especially for non-B. fragilis species. For non-Bacteroides anaerobes no major discrepancies were noted for Prevotella species, Peptostreptococcus species, and Viellonella parvula. With Clostridium species and Eubacterium species, some differences were noted with ceftizoxime because of differences in cut-off points. These data illustrate the magnitude of differences in results produced by the two methods using essentially the same test medium for the B. fragilis group. Fortunately, such major discordant results were not widely noted with other groups of anaerobes.

Increased in vitro activity of ceftriaxone by addition of tazobactam against clinical isolates of anaerobes.

A total of 461 clinical strains of anaerobes were tested using a broth microdilution test to determine the activity of the combination of ceftriaxone and tazobactam and other antimicrobials against these isolates. Ceftriaxone was combined with tazobactam in ratios of 1:1, 2:1, 4:1, and 8:1 and twofold dilutions of ceftriaxone in constant concentrations to tazobactam of 2, 4, 8, 16, and 32 micrograms/ml. Against beta-lactamase-producing strains of the Bacteroides fragilis group, B. capillosus, and Prevotella species all combinations of ceftriaxone and tazobactam showed enhanced in vitro activity and were eight- to 2048-fold more active than ceftriaxone alone. By comparison ceftriaxone and tazobactam showed superior or equal activity to ampicillin and sulbactam, piperacillin and tazobactam, amoxicillin and clavulanate, ticarcillin and clavulanate, and metronidazole against these same strains. Against beta-lactamase nonproducing strains of Porphyromonas, Fusobacterium, Clostridium, Eubacterium, Peptostreptococcus, and Veillonella parvula the addition of tazobactam produced no appreciable enhanced ceftriaxone activity. Fixed concentrations of tazobactam at 2 and 4 micrograms/ml appear to be most suitable for susceptibility testing and are within the pharmacologic profile of this inhibitor. Pharmacologic and toxicity studies will be needed to define the role of ceftriaxone and tazobactam in infectious diseases.

In vitro activity of biapenem (L-627), a new carbapenem, against anaerobes.

We tested 441 clinical strains of anaerobes by using a broth microdilution method to determine the in vitro activity of biapenem for comparison with those of other agents. Biapenem had activity comparable to those of imipenem and meropenem against all groups of anaerobes with MICs for 90% of the strains tested of 0.06 to 2 micrograms/ml. Against the Bacteroides fragilis group, biapenem was more active than ampicillin-sulbactam, ticarcillin-clavulanate, piperacillin, cefoxitin, cefotaxime, and ceftriaxone. Biapenem was also active against all of the B. capillosus, Prevotella, Clostridium, and Eubacterium strains and anaerobic cocci tested. Against all of the anaerobes tested, biapenem was 32- and 4-fold more active than clindamycin and metronidazole, respectively. These data indicate broad-spectrum activity by biapenem against anaerobes.

A five-year multicenter study of the susceptibility of the Bacteroides fragilis group isolates to cephalosporins, cephamins, penicillins, clindamycin, and metronidazole in the United States.

Over 2800 clinical strains of the Bacteroides fragilis group were collected during a 5-year period from ten geographically separate sites and tested for their susceptibility to various antimicrobial agents using a broth microdilution method. Among the cephalosporins, ceftizoxime was the most active (13% resistance) and importantly exhibited relatively equal activity against both B. fragilis species and non-B. fragilis species. Cefotaxime exhibited similar activity with an overall resistance rate of 18%. Both ceftriaxone and cefoperazone were appreciably less active cephalosporins especially against non-B. fragilis species. With regard to cephamycins, cefoxitin (MIC90, 32 micrograms/ml) was more active than cefotetan (MIC90, > or = 256 micrograms/ml) and cefmetazole (MIC90, 64 micrograms/ml). Non-B. fragilis species were highly resistant to cefotetan and cefmetazole. Imipenem was highly active against all strains with the exception of four strains of B. fragilis. Ampicillin-sulbactam, amoxicillin-clavulanate, piperacillin-tazobactam, and cefoperazone-sulbactam were all highly active with resistance rates < 2%. No resistance was detected to metronidazole, whereas 14% of isolates were resistant to clindamycin. When compared with other studies, these findings underscore the wide variability in susceptibility patterns reported nationwide and the need to continue monitoring these patterns to aid in choosing the most active compounds for therapy.

Anaerobic susceptibility testing. Slight differences in inoculum size can make a difference in minimum inhibitory concentrations.

In the present study, we compared the actual inoculum density from inoculated broth microdilution wells to the targeted inoculum size (10(5) CFU/well) when the inoculum was prepared using a McFarland nephelometer to achieve the standard density. Three target inoculum sizes (10(5), 5 x 10(5), and 10(6) CFU/well) were used to compare the effect of slight inoculum size increases of both ATCC and clinical strains of anaerobes on MICs of various antimicrobials. Actual colony counts of Bacteroides fragilis, Bacteroides thetaiotaomicron, Eubacterium lentum, and Veillonella parvula ranged from 0.7 x 10(5) to 1.4 x 10(5) CFU/well. As the inoculum size rose above the desired 10(5) CFU/well level, the MICs of certain antimicrobials became elevated. Ceftizoxime, cefotaxime, and ceftriaxone MICs rose 4- to 16-fold with as little as 0.5 log10 increase in inoculum size. Other increases were also noted with E. lentum and Clostridium perfringens, but were primarily between the low and high inoculum sizes. Results with cefoxitin, cefotetan, mezlocillin, and imipenem did not show an appreciable increased inoculum effect. This study demonstrates that the variation in organism size among anaerobes (both between species and within species) does make a difference in actual inoculum size and certain anaerobes may require special adjustment to ensure proper MIC results from susceptibility testing.

The rapid emergence of fluoroquinolone-methicillin-resistant Staphylococcus aureus infections in a community hospital. An in vitro look at alternative antimicrobial agents.

The introduction of ciprofloxacin on an unrestricted basis into a 900-bed community hospital resulted in the emergence of high-level fluoroquinolone resistance among methicillin-resistant Staphylococcus aureus (MRSA) during the subsequent 18 months. Susceptibility testing revealed several old and new compounds to which all the S. aureus strains were susceptible. When an MRSA strain became resistant to ciprofloxacin it also exhibited high-level resistance to ofloxacin, fleroxacin, norfloxacin, and enoxacin. Two new experimental fluoroquinolones, WIN 57273 and CI-960, exhibited good activity against all test strains. Among the glycopeptide compounds, mupirocin and teicoplanin were approximately fourfold more active than vancomycin and ramoplanin. Rifampin and trimethoprim-sulfamethoxazole (TMP/SMZ) showed good activity against most strains as did imipenem. For clindamycin, gentamicin, and tetracycline susceptibilities exhibited a bimodal distribution with at least 10% of strains having resistant MIC values. Surprisingly, the addition of sulbactam potentiated the activity of ampicillin against the ciprofloxacin-resistant MRSA strains, however, sulbactam had little effect on cefoperazone activity against these same strains. Time-kill kinetic studies of selected antimicrobials against ciprofloxacin-resistant strains indicated good killing by vancomycin, ampicillin-sulbactam, and TMP/SMZ. Teicoplanin was less bactericidal than vancomycin while these same strains rapidly developed resistance to rifampin even at concentrations 8 x MIC. These data indicate certain alternative compounds within our study warrant further investigation, especially in vivo, against multiply-resistant staphylococci.

In vitro antistaphylococcal activity and testing of RP 59500, a new streptogramin, by two methods.

The in vitro antistaphylococcal activity of RP 59500, a new streptogramin, was comparable to those of vancomycin and teicoplanin against Staphylococcus aureus, and RP 59500 was the most active agent against coagulase-negative staphylococci. All staphylococcal strains were inhibited by 4 micrograms of RP 59500 per ml, including multiply resistant strains. Broth microdilution and agar dilution testing gave comparable results in 97% of the tests with RP 59500.

Discordant results between the broth disk elution and broth microdilution susceptibility tests with Bacteroides fragilis group isolates.

Susceptibility testing of 161 clinical isolates of the Bacteroides fragilis group was performed to compare interpretive results generated by the broth disk elution and broth microdilution methods recommended by the National Committee for Clinical Laboratory Standards. Among the cephalosporin-cephamycin compounds tested, correlation was poorest for ceftizoxime (71%), ceftriaxone (57%), and cefotaxime (47%); when the tests did not correlate, false resistance was seen 92, 95, and 93% of the time, respectively. Cefotetan and cefoperazone showed lack of correlation in 19 and 20% of the tests, respectively. For cefotetan, false resistance was more frequent, while with cefoperazone, false susceptibility occurred more often. Cefoxitin produced the fewest discrepancies; 10% of the disk elution tests produced either false-resistance or false-susceptibility results. Mezlocillin and piperacillin showed lack of correlation in 8 and 14% of the tests, respectively, and discrepancies were due primarily to false-resistance results. Overall with the beta-lactams, 84% of the discordant interpretive results were false resistance by the broth disk elution test. Clindamycin had a discrepancy rate of 10%, with the majority of discrepancies being false susceptibility disk elution results. Because of the high number of discrepancies noted with ceftizoxime, ceftriaxone, and cefotaxime, we recommend that these drugs not be tested by the disk elution method and that they be tested by a quantitative MIC method such as the broth microdilution test. Furthermore, caution should be exercised when interpreting broth disk elution results with all the beta-lactams included in this study except imipenem. These data indicate the lack of correlation of results between these two tests for many beta-lactams and suggest the need for a reexamination of the disk elution method to provide a more accurately standardized test.

Lomefloxacin, a new fluoroquinolone. Studies on in vitro antimicrobial spectrum, potency, and development of resistance.

Lomefloxacin (NY-198; SC-47111), a potent new difluoroquinolone, was studied to compare its in vitro activity with that of other antimicrobials against 2194 clinical isolates. Lomefloxacin showed excellent inhibitory and bactericidal activity against strains of Enterobacteriaceae and inhibited greater than 99% of the isolates at a concentration of 4 micrograms/ml or less. Lomefloxacin exhibited good-to-moderate activity against strains of Acinetobacter (MIC90 4 micrograms/ml) and Pseudomonas aeruginosa (MIC90 8 micrograms/ml), but poor activity for Pseudomonas cepacia (MIC90 greater than 16 micrograms/ml). Staphylococcus aureus, and Staphylococcus epidermidis isolates, both oxacillin-susceptible and -resistant strains, were susceptible (MIC90 1 micrograms/ml) to lomefloxacin and the other fluoroquinolones. Strains of Haemophilus influenzae, (MIC90 less than or equal to 0.13 micrograms/ml) Neisseria gonorrhoeae (MIC90 less than or equal to 0.03 micrograms/ml), and Branhamella catarrhalis (MIC90 less than or equal to 0.03 micrograms/ml) were highly susceptible to lomefloxacin. Streptococcal isolates, especially viridans streptococci, were considerably less susceptible to the fluoroquinolones. Overall, lomefloxacin had comparable activity to norfloxacin, fleroxacin, and ofloxacin, and against many facultative anaerobes lomefloxacin was more active than imipenem, cefotaxime, ceftazidime, ticarcillin/clavulanic acid, aztreonam, trimethoprim/sulfamethoxazole and gentamicin. Development of resistance to lomefloxacin by spontaneous mutation was low and comparable to that of other fluoroquinolones. Growth in subinhibitory concentrations resulted in increased resistance to fluoroquinolones for selected test strains.

Comparative in vitro activity of the two new oral cephalosporin metabolites RO 19-5247 and RO 15-8074.

A total of 629 clinical strains of gram positive and gram negative bacteria were tested for their susceptibility to RO 19-5247, RO 15-8074, and other antimicrobial agents. Both RO 19-5247 and RO 15-8074 had good activity against strains of Enterobacteriaceae; however, resistance was found among some strains of Enterobacter, Citrobacter, Klebsiella and Morganella spp. Both compounds showed moderate to poor active against Acinetobacter spp., Pseudomonas aeruginosa, staphylococci and Streptococcus faecalis. Against strains of Haemophilus influenzae, Neisseria gonorrhoeae, Gardnerella vaginalis, Streptococcus pneumoniae and streptococci (not enterococci), each compound was highly active in vitro. RO 19-5247 and RO 15-8074 had comparable activity to cotrimoxazole, ceftazidime and ceftizoxime. Each new compound had considerably better activity then did cefaclor and amoxicillin/potassium clavulanate.

RO23-6240, a new orally absorbed quinolone: in vitro comparison with other broad-spectrum oral antimicrobial agents and imipenem.

A total of 626 clinical isolates were tested for their susceptibility to RO23-6240 and other broad-spectrum antimicrobial agents. RO23-6240 showed good activity against strains of Enterobacteriaceae, Acinetobacter, and P. aeruginosa. RO23-6240 MIC90s ranged from 0.032 to 4 micrograms/ml for these strains. RO23-6240 also showed good activity against staphylococci, both methicillin-susceptible and -resistant strains. The activity in vitro of RO23-6240 was comparable with that of norfloxacin and ofloxacin, and more active than imipenem, trimethoprim-sulfamethoxazole, cefaclor, and amoxycillin/clavulanate potassium. As with the other quinolones tested, increases in inoculum size produced moderate increases in the MICs and MBCs of RO23-6240.

Comparative in vitro activity of the new oral cephalosporin BMY-28100.

Using a broth microdilution method, the in vitro activity of BMY-28100 against 365 clinical strains of commonly isolated bacteria was determined. BMY-28100 showed good activity against streptococci, methicillin-susceptible staphylococci, Salmonella spp., Shigella spp., and beta-lactamase producing Branhamella catarrhalis and Haemophilus influenzae. Against susceptible strains of these organisms, BMY-28100 showed activity comparable to that of penicillin G, ampicillin, co-trimoxazole, erythromycin, cefaclor, doxycycline and amoxicillin/potassium clavulanate. BMY-28100 had moderate activity against Arizona hinshawii and poor activity against Campylobacter jejuni and Yersinia enterocolitica.