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Azithromycin and clarithromycin are both highly active in vitro against Gram-positive respiratory pathogens, but azithromycin is substantially more potent against Haemophilus influenzae. We investigated the susceptibility of H. influenzae, Moraxella catarrhalis, Streptococcus pneumoniae and S. pyogenes to azithromycin and clarithromycin, and determined the prevalence of beta-lactamase production in H. influenzae and M. catarrhalis. Results from three geographic regions of the USA were compared. A significantly greater proportion of H. influenzae isolates were susceptible to azithromycin than to clarithromycin (95.7% vs 63.1%, P < 0.001). M. catarrhalis, S. pneumoniae and S. pyogenes were highly susceptible to both antibiotics with no significant differences in susceptibility. beta-Lactamase was produced by 94.8% of M. catarrhalis isolates and 35.6% of H. influenzae.

E-test, broth microdilution, and agar dilution susceptibility tests were used to evaluate penicillin and five extended-spectrum cephalosporins against 196 strains of Streptococcus pneumoniae with different levels of penicillin resistance. Oxacillin disk tests corresponded to minimum inhibitory concentrations of penicillin determined by the E-test better than those generated by broth microdilution or agar dilution methods. Relative potency of the study drugs was as follows: cefotaxime = ceftriaxone > penicillin > cefuroxime > ceftizoxime > ceftazidime.

Over 2,000 clinical isolates from ten American medical centers were tested for susceptibility to cefotaxine, ceftriaxone, ceftizoxime, and ceftazidime by both broth microdilution and disk diffusion methods. Typically resistant (e.g. enterococci) and highly susceptible (e.g. streptococci) isolates showed no change in susceptibility patterns compared to previous surveys. Pseudomonas aeruginosa and Stenotrophomonas maltophilia exhibited significant decreases in susceptibility to these cephalosporins. Among Escherichia coli and Klebsiella spp isolates, two to three percent were resistant to one of the four drugs whereas they were very susceptible to the other three. Of the four antibiotics, the cefotaxime disk diffusion test results correlated best with the microdilution results. With the other three drugs the disk diffusion test yielded 1 to 9% more susceptible test results and 1 to 20% fewer resistant test results than broth microdilution when testing gram negative bacilli. The clinical significance of such discrepancies is not known, but the impact on antibiotic susceptibility surveys and antibiogram comparisons could be significant.

The l-isomer of ofloxacin (levofloxacin) was twice as active as ofloxacin and the d-isomer was inactive against 654 Streptococcus pneumoniae isolates. For disc susceptibility tests of pneumococci, 5 micrograms levofloxacin discs can be used with interpretive criteria of < or = 12 mm for resistant (MIC > or = 8.0 mg/L) and > or = 16 mm for susceptible (MIC < or = 2.0 mg/L). For quality control of levofloxacin tests, S. pneumoniae ATCC 49619 should give zones 20-25 mm in diameter and MICs 0.5-2.0 mg/L.

The susceptibility of Neisseria gonorrhoeae to PD 131628 and grepafloxacin (OPC 17116) was evaluated by agar dilution and disc diffusion methods. A tentative susceptibility category for both fluoroquinolones included strains for which the MICs are < or = 0.06 mg/L and the zones of inhibition are > or = 38 mm for PD 131628 and > or = 37 mm for grepafloxacin. Quality control studies with N. gonorrhoeae ATCC 49226 suggested that agar dilution MIC limits were 0.002-0.008 mg/L and 0.004-0.03 mg/L for PD 131628 and grepafloxacin, respectively. The zone size limits were 50-58 mm for PD 131628 and 44-52 mm for grepafloxacin.

Variation in the DNA profile and antifungal susceptibility of 271 bloodstream isolates of Candida species was evaluated. The isolates were obtained from 152 nonneutropenic patients hospitalized in 21 different medical centers. In general, each patient was infected with their own distinct DNA type of Candida, and in those cases in which multiple cultures were positive, the same strain was isolated repeatedly over time. Minimum inhibitory concentrations of fluconazole or amphotericin B did not increase over time among the strains isolated repeatedly from individual patients.

Proposed quality control (QC) parameters for susceptibility testing of Streptococcus pneumoniae to cefdinir were developed following the procedure recommended by the National Committee for Clinical Laboratory Standards. The proposed QC MIC range for microdilution susceptibility testing of S. pneumoniae ATCC 49619 is 0.06-0.25 mg/L. The proposed QC limits for inhibitory zone diameters of S. pneumoniae ATCC 49619 around 5 micrograms cefdinir disks is 26-31 mm. We recommend the following for tentative interpretive criteria for determining the susceptibility of S. pneumoniae to cefdinir: susceptible, MIC < or = 0.5 mg/L or inhibition zone diameter > or = 23 mm; intermediate, MIC 1.0 mg/L or inhibition zone, 20-22 mm; resistant, MIC > or = 2.0 mg/L or inhibition zone diameter, < or = 19 mm for broth microdilution and disc diffusion tests, respectively.

The in vitro activities of four extended-spectrum cephalosporins and benzyl penicillin were evaluated against 698 clinical isolates of Streptococcus pneumoniae, including 130 (19%) penicillin-intermediate and 84 (12%) penicillin-resistant strains. Cefotaxime and ceftriaxone were essentially identical in their antipneumococcal activities: both were active against penicillin-susceptible strains and most penicillin-intermediate strains. Cefpirome was twice as potent as cefotaxime and ceftriaxone against penicillin-resistant strains. Ceftazidime was 8- to 16-fold less active than cefotaxime and ceftriaxone against S. pneumoniae in vitro, and thus, its spectrum included only penicillin-susceptible strains.

In vitro susceptibility tests were performed with 350 selected strains of Streptococcus pneumoniae to evaluate disk diffusion tests with 30 micrograms and 1 microgram cefotaxime disks. Zones were compared to MICs of cefotaxime with and without its desacetyl metabolite. Cefotaxime was two to eight times more active than desacetyl cefotaxime, but the two compounds were additive when combined in vitro. For 30 micrograms disks, zone size breakpoints were < or = 27 mm, 28-30 mm and > or = 31 mm for resistant, intermediate and susceptible, respectively. For 1 microgram disks, those zone size criteria were reduced to < or = 13 mm, 14-16 mm and > or = 17 mm. The 30 micrograms disk that is currently available for testing other species can be used for testing pneumococci; however, the 1 microgram disk has some important advantages.

The in vitro antimicrobial activity of an oral penem, WY-49605, was compared with those of six other oral antimicrobial agents against 598 bacterial isolates representing 51 different species. WY-49605 exhibited good activity against most gram-positive bacteria and members of the family Enterobacteriaceae. It had little activity against nonfermenting gram-negative bacilli, Enterobacter spp., Serratia spp., enterococci, Staphylococcus haemolyticus, and methicillin-resistant Staphylococcus aureus. Its activity was unaffected by the beta-lactamases of Neisseria gonorrhoeae, Haemophilus influenzae, and staphylococci. Disk diffusion susceptibility tests were performed with 5-, 10-, 15-, and 30-micrograms WY-49605 disks. The 5-micrograms disk is recommended, with tentative breakpoints of > or = 16 mm for susceptibility (MIC, < or = 2.0 microgram/ml) and < or = 12 mm for resistance (MIC, > or = 8.0 micrograms/ml).

Among 698 Streptococcus pneumoniae isolates, 475 were penicillin susceptible and > 99% of those were susceptible to 0.5 microgram of cefixime per ml; other pneumococci were tentatively assumed to be resistant to cefixime. A 1-microgram oxacillin disk was more reliable than a 5-micrograms cefixime disk for predicting susceptibility to cefixime.

Grepafloxacin (OPC 17116) and PD 131628 were evaluated against 150 Haemophilus influenzae isolates to propose susceptibility testing criteria for both broth dilution and disk diffusion procedures using haemophilus test medium. Grepafloxacin-susceptible isolates are defined as those for which minimum inhibitory concentrations (MICs) are < or = 0.06 micrograms/ml and zones of inhibition are > or = 27 mm. PD 131628-susceptible strains of H. influenzae included those that exhibited MICs < or = 0.03 micrograms/ml, and the zones were > or = 32 mm. All MICs were well below concentrations that can be achieved in the blood and tissues of patients treated with either study drug. Criteria for defining a resistant category cannot be determined until strains with elevated MICs are available for study. The proposed quality control guidelines for H. influenzae ATCC 49247 and the disk test are 32-39 mm in diameter for grepafloxacin and 34-42 mm for PD 131628. The preliminary broth microdilution MIC control limits for this strain are 0.002-0.016 micrograms/ml for grepafloxacin and 0.002-0.008 micrograms/ml for PD 131628.

For testing bacterial susceptibility to PD131628, a 5-micrograms disk and the following tentative interpretive criteria may be used: > or = 19 mm for susceptible (MIC, < or = 1.0 micrograms/ml), 16 to 18 mm for intermediate (MIC, 2.0 micrograms/ml), and < or = 15 mm for resistant (MIC, > or = 4.0 micrograms/ml). For standard quality control strains, the following limits are proposed: for Escherichia coli ATCC 25922, zones of 31 to 41 mm or a MIC of 0.002 to 0.016 micrograms/ml; for Pseudomonas aeruginosa ATCC 27853, zones of 26 to 34 mm or a MIC of 0.12 to 0.5 micrograms/ml; for Staphylococcus aureus ATCC 25923, zones of 27 to 33 mm; for Staphylococcus aureus ATCC 29213, a MIC of 0.03 to 0.12 micrograms/ml; and for Enterococcus faecalis ATCC 29212, a MIC of 0.12 to 0.5 micrograms/ml.

Broth microdilution tests were carried out with 2,671 respiratory tract isolates from 19 medical centers throughout the continental United States. The tests compared a streptogramin (RP59500) to erythromycin, dirithromycin, clarithromycin, and azithromycin against Streptococcus pneumoniae, S. pyogenes, Haemophilus influenzae, and Moraxella catarrhalis. Against macrolide-susceptible strains, the potency of RP59500 was similar to that of the macrolides: the azalide, azithromycin, was two to four times more potent against H. influenzae. The azalide and three macrolides showed nearly complete cross-resistance. At 2.0 micrograms/ml or less, the streptogramin, RP59500, was active against both macrolide-resistant and -susceptible strains of S. pneumoniae (MIC for 50% of the strains tested, 0.25 microgram/ml; MIC for 90% of the strains tested, 0.5 microgram/ml).

A comparative evaluation of the in vitro susceptibilities of 597 clinical yeast isolates to amphotericin B, fluconazole, and 5-fluorocytosine (5FC) was conducted. The broth macrodilution reference method of the National Committee for Clinical Laboratory Standards (NCCLS, M27-P) was adapted to the microdilution method. Microdilution endpoints for amphotericin B were scored as the lowest concentration in which a score of 0 (complete absence of growth) was observed and for 5FC and fluconazole as the lowest concentration in which a score of 2 (prominent decrease in turbidity; MIC-2) was observed compared to the growth control. The MIC values were read after 24 and 48 h incubation. A broad range of MIC values was observed with each antifungal agent. Amphotericin B was very active (MIC90 < or = 1.0 microgram/ml) against all of the yeast isolates with the exception of C. lusitaniae (MIC90 > or = 2.0 micrograms/ml). Fluconazole was most active against C. parapsilosis (MIC90 of 1.0 microgram/ml) and least active against C. krusei (MIC90 of 32 micrograms/ml). 5FC was most active against C. albicans, C. parapsilosis, C. tropicalis, and T. glabrata (MIC90 < or = 1.0 microgram/ml) and was least active against C. krusei and C. lusitaniae (MIC90 > or = 16 micrograms/ml). These data indicate that the microdilution method, performed in accordance with M27-P, provides a means of testing larger numbers of yeast isolates against an array of antifungal agents and allows this to be accomplished in a reproducible and standardized manner. Given these results, it appears that the microdilution method may be a useful alternative to the macrodilution reference method for susceptibility testing of yeasts.

The antifungal susceptibilities of 232 pathogenic blood stream Candida isolates collected during a recently completed trial comparing fluconazole (400 mg/day) with amphotericin B (0.5 mg/kg of body weight per day) as treatment for candidemia in the nonneutropenic patient were determined both by the National committee for Clinical Laboratory Standards M27-P macrobroth methodology and by a less cumbersome broth microdilution methodology. For amphotericin B, M27-P yielded a very narrow range of MICs (0.125 to 1 microgram/ml) and there were no susceptibility differences among species. For fluconazole, a broad range of MICs were seen (0.125 to > 64 micrograms/ml), with characteristic MICs seen for each species in the rank order Candida albicans < C. parapsilosis approximately equal to C. lusitaniae < C. glabrata approximately equal to C. krusei approximately equal to C. lipolytica. The MIC distribution for C. tropicalis was bimodal and could not be ranked. Both microdilution MICs were within one tube dilution of the M27-P MIC for > 90% of isolates with amphotericin B and for > or = 77% of isolates with fluconazole. For both methods, elevated MICs did not predict treatment failure. In the case of amphotericin B, the MIC range was too narrow to permit identification of resistant isolates. In the case of fluconazole, MICs for isolates associated with failure to clear the bloodstream consistently were equivalent to the median MIC for the given species. Successful courses of therapy were seen with four isolates from four patients despite MICs of > or = 32 micrograms/ml. As MICs obtained by M27-P and similar methods correlate with responsiveness to fluconazole therapy in animal models and in AIDS patients with oropharyngeal candidiasis, the lack of correlation in this setting suggests that the MICs for these isolates are at or below the relevant fluconazole breakpoint for this dose of fluconazole and patient setting and that host factors such as failure to exchange intravenous catheters were more important than MIC in predicting outcome.

Chemotherapy of community-acquired respiratory tract infections was reviewed from a microbiological perspective. The current worldwide spread of penicillin-resistant Streptococcus pneumoniae and of ampicillin-resistant Haemophilus influenzae has required a reassessment of the antimicrobial agents being used for empiric therapy. In vitro data with different orally administered antibiotics were reviewed in order to identify any deficiencies in their spectra of activity against four common respiratory tract pathogens. Cefixime, cefuroxime axetil, cefprozil, amoxicillin-clavulanic acid and trimethoprim-sulfamethoxazole were active against all four species other than penicillin-resistant pneumococci.

One hundred fifty isolates of Haemophilus influenzae (including 30 beta-lactamase-positive strains and 23 beta-lactamase-negative, ampicillin-resistant strains) were tested for susceptibilities to ampicillin, amoxicillin, and amoxicillin-clavulanic acid (A/C) by the broth microdilution method in Haemophilus test medium (HTM) and in Mueller-Hinton medium with lysed horse blood and by the disk diffusion method on HTM agar. Our results support the use of HTM for susceptibility testing of H. influenzae but raise a number of questions regarding the interpretive criteria currently in use, particularly with respect to the fourfold difference in MIC susceptibility breakpoints for ampicillin and A/C and a resulting high proportion of A/C-susceptible beta-lactamase-negative, ampicillin-resistant strains.

Clinical isolates of Haemophilus influenzae, Streptococcus pneumoniae, Streptococcus pyogenes, and Moraxella catarrhalis were gathered from 19 different clinical laboratories throughout the continental United States. The in vitro activities of 12 orally administered antimicrobial agents were compared by broth microdilution tests with 3,151 bacterial isolates. Among 890 H. influenzae isolates, 30% were capable of producing beta-lactamase enzymes (12 to 41% in different medical centers). Most of the 619 beta-lactamase-negative H. influenzae strains were susceptible to ampicillicin (MIC, < or = 1.0 micrograms/ml): 5 strains were intermediate in susceptibility (MIC, 2.0 micrograms/ml) and 1 strain was ampilicillin resistant (MIC, 4.0 micrograms/ml). Ninety-two percent of 698 M. catarrhalis strains were beta-lactamase positive. Of 799 S. pneumoniae isolates, 15% were intermediate in susceptibility to penicillin and 7% were resistant to penicillin. The prevalence of penicillin-susceptible pneumococci in different institutions ranged from 63 to 95%. Only 1% of 764 S. pyogenes isolates were resistant to the macrolides, but 5% of S. pneumoniae isolates were macrolide resistant. Only 71% of 58 penicillin-resistant S. pneumoniae isolates were erythromycin susceptible, whereas 97% of the 622 penicillin-susceptible strains were erythromycin susceptible. Penicillin-resistant pneumococci were also relatively resistant to the cephalosporins and amoxicillin. Penicillin-susceptible pneumococci were susceptible to amoxicillin-clavulanic acid (MIC for 90% of isolates tested [MIC90], < or = 0.12/0.06 microgram/ml), cefixime (MIC90, 0.25 microgram/ml), cefuroxime axetil (MIC90, < or = 0.5 microgram/ml), cefprozil (MIC90, < or = 0.5 micrograms/ml), cefaclor (MIC90, 0.5 microgram/ml), and loracarbef (MIC90, 1.0 microgram/ml). Most strains of the other species remained susceptible to the study drugs other than amoxicillin.

A comparison of the E test, the broth microdilution test, and the reference broth macrodilution susceptibility test of the National Committee for Clinical Laboratory Standards for fluconazole susceptibility testing was performed with 238 clinical isolates of Candida species and Torulopsis (Candida) glabrata. An 80% inhibition endpoint MIC was determined by the reference broth macrodilution method after 48 h of incubation. The MICs obtained by the two study methods were read after 24 and 48 h of incubation. Overall, excellent agreement within 2 doubling dilutions was obtained between the broth microdilution and the broth macrodilution methods for the combined results for all species at both 24 h (93%) and 48 h (94%). The correlation of 24-h MIC endpoints between the E test and the broth macrodilution methods was 37% for T. glabrata, 56% for Candida tropicalis, 93% for Candida albicans, and 90% for other Candida species. The percent agreement at 48 h ranged from 34% for T. glabrata to 97% for Candida species other than C. albicans and C. tropicalis. These initial results support the further evaluation of the E test as an alternative method for fluconazole susceptibility testing of Candida species.

A comparative evaluation of two broth microdilution methods for antifungal susceptibility testing of 600 clinical yeast isolates (Candida spp., Torulopsis glabrata, and Cryptococcus neoformans) against amphotericin B, fluconazole, and flucytosine (5FC) was conducted. Microdilution testing was performed according to National Committee for Clinical Laboratory Standards (NCCLS) recommendations (NCCLS document M27-P). By using the growth control for comparison, reference microdilution MIC endpoints for amphotericin B were scored as the lowest concentration at which a score of 0 (complete absence of growth) was observed, and those for 5FC and fluconazole were scored at the lowest concentration at which a score of 2 (prominent decrease in turbidity) (MIC-2) was observed. The second microdilution method employed a colorimetric endpoint using an oxidation-reduction indicator (Alamar Biosciences, Inc., Sacramento, Calif.) and was assessed independently of the reference microdilution MICs. The MICs for the two microdilution test systems were read after 24 and 48 h of incubation. Excellent agreement between the reference and colorimetric microdilution MICs was observed. Overall agreement was > or = 95% for all three drugs at 24 h. At 48 h, agreement was > or = 98% for amphotericin B and 5FC but dropped to 84% for fluconazole. Given these results it appears that the colorimetric microdilution approach to antifungal susceptibility testing may be viable alternative to the NCCLS reference method for testing yeasts.

The antibacterial activity of PD 131628 was compared with that of ciprofloxacin against 401 clinical isolates. The two drugs had comparable MICs for most Gram-negative species. PD 131628 was two to eight times more active against Gram-positive species and against Xanthomonas maltophilia. Both drugs were bactericidal, Among Gram-positive isolates, resistant mutants were not detected in vitro. Among Gram-negative bacilli, spontaneously occurring mutants resistant to PD 131628 were readily demonstrated more frequently than for ciprofloxacin. Resistance to PD 131628 was more readily induced by serially transferring the Gram-negative species, and resistant organisms emerged more rapidly on prolonged incubation in time-kill studies. Correlating the PD 131628 5 micrograms disc diffusion zone diameters with PD 131628 MICs, the following breakpoints are tentatively proposed: susceptible, MIC < or = 1 mg/L or zone > or = 19 mm; intermediate, MIC 2.0 mg/L or zones 16-18 mm; and resistant, MIC > or = 4.0 mg/L or zones < or = 15 mm.

Susceptibility tests were performed on 2402 of consecutive isolates of Enterobacteriaceae and 254 strains of Pseudomonas aeruginosa in five separate medical centres. Five beta-lactams were evaluated with and without a beta-lactamase inhibitor. The effect of different inhibitors was species specific and the rank order of decreasing efficacy against all enteric bacilli was: cefoperazone-sulbactam > piperacillin-tazobactam > cefoperazone > ticarcillin-clavulanic acid > piperacillin > co-amoxiclav > ampicillin-sulbactam > ticarcillin > ampicillin > amoxycillin.

The in vitro activity of ceftriaxone and six additional antimicrobial agents (ceftizoxime, cefoperazone, cefuroxime, fleroxacin, ciprofloxacin, and trimethoprim/sulfamethoxazole) was assessed or 602 recent clinical isolates of staphylococci from six geographically distinct medical centers in North America. All seven antimicrobial agents were active (90-100% of strains susceptible) against oxacillin-susceptible (OS) strains of Staphylococcus aureus (OSSA) and coagulase-negative staphylococci (OSCNS) but had limited activity against oxacillin resistant (OR) staphylococci. Our assessment of the in vitro antistaphylococcal activity of ceftriaxone against contemporary isolates of Staphylococcus aureus and coagulase-negative staphylococci indicates that the activity versus OS staphylococci has not changed over the past decade despite widespread use of the drug. It appears that these agents will continue to be useful for empiric therapy in those centers in which OR strains are uncommon.

Sparfloxacin and levofloxacin were evaluated against 150 Haemophilus influenzae isolates and 149 Neisseria gonorrhoeae isolates in order to define susceptibility testing parameters. Sparfloxacin-susceptible H. influenzae strains were defined as those for which the MICs were < or = 0.25 microgram/ml and the zones were > or = 30 mm, and N. gonorrhoeae susceptible strains were those for which the MICs were < or = 0.03 microgram/ml and the zones were > or = 39 mm (5-micrograms disks). Levofloxacin-susceptible strains of H. influenzae included those for which the MICs were < or = 0.12 microgram/ml and the zones were > or = 32 mm and N. gonorrhoeae susceptible strains were those for which the MICs were < or = 0.12 microgram/ml and the zones were > or = 37 mm (5-micrograms disks). Criteria for a resistant category cannot yet be defined for either quinolone. In multilaboratory studies with different lots of Haemophilus Test Medium, replicate tests with the standard control strain of H. influenzae (ATCC 49247) were evaluated. For sparfloxacin disk tests, the proposed zone size limits were 33 to 42 mm and broth microdilution MIC limits were 0.004 to 0.016 microgram/ml, whereas for levofloxacin tests, zone size limits were 32 to 41 mm and broth microdilution MIC limits were 0.008 to 0.03 microgram/ml. Other multilaboratory studies evaluated tests with supplemented GC agar and N. gonorrhoeae ATCC 49226; for both drugs, zone size limits were 44 to 52 mm and agar dilution MIC limits were 0.004 to 0.016 microgram/ml.

Ampicillin was generally twice as active as amoxicillin against 2440 consecutive isolates of Enterobacteriaceae from five medical centers. When beta-lactamase inhibitors were added to the penicillins, there was a significant increase in susceptibility. The magnitude of the increased susceptibility to ampicillin-sulbactam (A-S) and amoxicillin-clavulanic (A-C) acid varied with the species and types of beta-lactamases elaborated. Although cross-susceptibility and cross-resistance between ampicillin and amoxicillin was nearly complete, major differences were documented between A-S and A-C with 6.7% of our consecutive isolates of Enterobacteriaceae. The clinical significance of these findings remains uncertain, but they may help explain some of the discrepancies occasionally observed by clinical microbiologists with the combination drugs.

Susceptibility to sparfloxacin was tested simultaneously by the agar dilution and disk diffusion methods. For tests with 5-micrograms sparfloxacin disks, we propose that the breakpoints for the susceptibility and resistance categories should be > or = 19 and < or = 15 mm, respectively. Most minor discrepancies involved tests with pseudomonads.

Fosfomycin tromethamine is an orally administered fosfomycin that may be used for single-dose therapy of uncomplicated urinary tract infections. At breakpoint concentrations [< or = 128 micrograms/ml plus 25 micrograms/ml glucose-6-phosphate (G-6-P)], fosfomycin tromethamine inhibited > 90% of the 350 bacterial isolates tested. When testing Escherichia coli, Klebsiella spp., and Enterobacter spp., we note that the performance of fosfomycin disks improved when G-6-P was added to the disks. The interpretive error rates were minimized when 200-micrograms fosfomycin disks were supplemented with either 50 or 100 micrograms G-6-P. Using < or = 128 and > or = 256 micrograms/ml as the susceptible and resistant MIC breakpoints, respectively, the regression-analysis-derived disk diffusion zone diameter breakpoints for the 200-micrograms fosfomycin disk supplemented with 50 micrograms of G-6-P are as follows: susceptible, > or = 16 mm; intermediate, 13-15 mm; and resistant, < or = 12 mm.

Studies were undertaken to determine whether broth microdilution susceptibility tests could be standardized by using a single medium for testing fastidious respiratory pathogens. Mueller-Hinton broth with lysed horse blood and the broth version of Haemophilus Test Medium (HTM) were directly compared. Ten orally administered agents were found to give essentially identical results in both media but minor differences were noted. Because the test are easier to read when HTM broth is used, that medium is to be preferred for routine testing of Haemophilus influenzae, Streptococcus pneumoniae, Streptococcus pyogenes and Moraxella catarrhalis isolates by the microdilution procedure.

Levofloxacin disk susceptibility test criteria were evaluated by testing 350 bacterial isolates. Either 5- or 10-micrograms disks could be used satisfactorily. A 5-micrograms levofloxacin disk with zone size breakpoints of < or = 12 mm for resistance (MIC, > or = 8.0 micrograms/ml) and > or = 16 mm for susceptibility (MIC, < or = 2.0 micrograms/ml) is recommended.

Ramoplanin and mupirocin are two antimicrobial agents that may be applied topically. They had similar potencies against susceptible gram-positive cocci, but they differed in their spectra of activity. In plastic microdilution trays, the in vitro activity of ramoplanin was slightly diminished but that activity was restored by the addition of bovine serum albumin (0.02%). Susceptible strains were defined as those for which MICs of ramoplanin were < or = 2.0 micrograms/ml (without bovine serum albumin) and which had > or = 11-mm zones around 60-micrograms ramoplanin disks. Criteria for a resistant category cannot be defined at this time.

The susceptibility of anaerobic bacteria to two new fluoroquinolones, PD 131628 (the bioactive form of PD 131112 or CI-990) and clinafloxacin (CI-960 or PD 127391), was determined with the agar dilution procedures and two media, and one broth microdilution procedure. Sparfloxacin and ciprofloxacin were also tested by the broth microdilution method. One hundred anaerobic isolates and four control strains were tested by the three methods which gave minimum inhibitory concentrations (MICs) that were essentially comparable, but not identical. With the broth microdilution method, the relative potency of the four fluoroquinolones was: clinafloxacin > PD 131628 > sparfloxacin > ciprofloxacin. For the latter three drugs but not clinafloxacin, the MIC values were often near the proposed interpretive breakpoint concentrations, and thus minor methodological differences frequently influenced the interpretive categories. Replicate agar dilution tests in five laboratories established MIC control limits for each of four control strains: those MIC limits could also be used to define the expected performance of the two alternative methods.

Ampicillin combined with sulbactam was tested at both fixed ratio (2:1 and 1:1) and fixed sulbactam concentrations (4 micrograms/ml, 8 micrograms/ml and 16 micrograms/ml) against 2440 consecutively isolated gram-negative bacilli. Sulbactam significantly enhanced the spectrum of ampicillin activity. Overall, at 8 micrograms/ml ampicillin inhibited 50% of the Enterobacteriaceae isolates, whereas 69% to 84% of the isolates were inhibited by the various sulbactam combinations. The widest spectrum of activity for ampicillin/sulbactam was achieved by testing at a fixed sulbactam concentration of 16 micrograms/ml, followed by the 1:1 ratio and the fixed 8 micrograms/ml (84%, 76% and 74% inhibited, respectively). The amount of sulbactam at the susceptible breakpoint concentrations of ampicillin markedly affected the percentage of susceptible strains. Combinations that include 8 micrograms/ml of sulbactam are suggested for consideration.

Studies with fosfomycin tromethamine disks containing 200 micrograms of fosfomycin and 50 micrograms of glucose-6-phosphate confirmed the following zone diameter criteria for the NCCLS method: < or = 12 mm for resistant (MIC > or = 256 micrograms/ml), 13-15 mm for intermediate (MIC 128 micrograms/ml) and > or = 16 mm for susceptible (MIC < or = 64 micrograms/ml). Additional studies defined acceptable MIC and zone diameter ranges for the following quality control strains: Escherichia coli ATCC 25922, MIC 0.5 to 4.0 micrograms/ml, zone diameter 23 to 29 mm; Staphylococcus aureus ATCC 25923, zone diameter 26 to 32 mm; Pseudomonas aeruginosa ATCC 27813, MIC 2.0 to 8.0 micrograms/ml; and Enterococcus faecalis, ATCC 29212, MIC 16 to 64 micrograms/ml.

Ro 40-6890, the active metabolite of Ro 41-3399, was tested against 391 gram-negative and gram-positive clinical isolates. Ro 40-6890 was active against members of the family Enterobacteriaceae, Moraxella catarrhalis, pneumococci, other Streptococcus spp., and methicillin-susceptible staphylococci. Preliminary disk diffusion interpretive zone criteria were calculated for 5-, 10-, and 30-micrograms Ro 40-6890 disks and several possible MIC susceptibility breakpoints. We recommend the use of the 5-micrograms disk and suggest that the following zone diameters be used as breakpoints in clinical trials: susceptible, > or = 21 mm (MIC, < or = 1 microgram/ml); intermediate, 18 to 20 mm (MIC, 2 micrograms/ml); and resistant, < or = 17 mm (MIC, > or = 4 micrograms/ml).

A study was conducted to select a fluoroquinolone disk that could be used to represent other members of the fluoroquinolone class of drugs for routine susceptibility testing in the laboratory. Eight different fluoroquinolone agents were tested by disk diffusion and broth microdilution methods using 185 bacterial isolates, including 60 Enterobacteriaceae, 50 Pseudomonas species, 45 other gram-negative bacilli, and 30 Staphylococcus aureus. False-susceptible and false-resistant test results occurred at rates of 0-10% when disks containing different agents were tested against this unusually resistant collection of isolates. Much lower error rates would be expected with the type of microorganisms routinely encountered in clinical laboratories. Pefloxacin, enoxacin, or ciprofloxacin disks were not useful as class representatives. However, amifloxacin, lomefloxacin, ofloxacin, temafloxacin, or fleroxacin disks could be used to represent the other fluoroquinolones for routine susceptibility testing with a minimal chance of incurring serious errors.

Dirithromycin and its bioactive metabolite (erythromycylamine) were compared in vitro to erythromycin against 450 bacterial isolates. Dirithromycin and erythromycylamine did not differ in their in-vitro activity and the two compounds were additive when combined in equal proportions. Both compounds were active against erythromycin-susceptible bacteria but erythromycin was two to four times more potent on a weight-to-weight basis. Interpretive criteria for dirithromycin were not based on achievable blood concentrations since tissue concentrations may be 20-30 times greater than peak serum levels. Dirithromycin-susceptible strains were those with zones > or = 19 mm in diameter (15 micrograms dirithromycin disc) or MIC < or = 2.0 mg/L and resistance was defined as a zone < or = 15 mm or MIC > or = 8.0 mg/L. Interpretive criteria for testing Haemophilus influenzae on Haemophilus Test Medium (HTM) agar were not defined because of the drug's poor activity in the test system used. Two multi-laboratory studies were also undertaken, one to define quality control limits for disc diffusion tests and the other to define broth microdilution MIC control limits for tests with dirithromycin.

In vitro studies evaluated methods for testing the susceptibility of Haemophilus influenzae to piperacillin-tazobactam combinations. Ampicillin-resistant beta-lactamase-nonproducing strains of H. influenzae may be presumed to be relatively resistant to combinations of piperacillin-tazobactam, even though they frequently appear to be susceptible by disk diffusion methods. Other ampicillin-resistant or -susceptible strains were predictably susceptible; i.e., 130 such strains gave zones of inhibition > or = 26 mm in diameter, and MICs for these strains were < or = 0.125/4.0 micrograms/ml (< or = 1.0/0.12 micrograms/ml when an 8:1 ratio was tested). A resistant category has yet to be defined. For quality control purposes, H. influenzae ATCC 49247 should give zones of inhibition 32 to 38 mm in diameter, and broth microdilution MICs should be 0.12/4.0 to 0.5/4.0 micrograms/ml.

In in vitro studies we evaluated the susceptibilities of beta-lactamase-producing and -nonproducing, ampicillin-resistant strains of Haemophilus influenzae and compared them with those of ampicillin-susceptible strains. Ampicillin, amoxicillin-clavulanic acid, ceftibuten, cefaclor, cefuroxime, cefixime, and cefotaxime were evaluated by broth microdilution tests and disk diffusion tests. The disk diffusion tests accurately categorized beta-lactamase-producing strains and ampicillin-susceptible strains as being susceptible to the study drugs other than ampicillin. Ampicillin-resistant, beta-lactamase-nonproducing strains were relatively resistant to all seven study drugs, but the disk diffusion test did not always predict that resistance. The clinical relevance of the decreased susceptibility to various agents remains unclear, but to be conservative, all ampicillin-resistant, beta-lactamase-nonproducing strains might be assumed to be resistant to other beta-lactams. After excluding that small group of isolates, reliable susceptibility test results were obtained with lots of Haemophilus Test Medium that met quality assurance criteria.

The susceptibility of 410 clinical bacterial isolates to RO 23-9424, a novel cephalosporin 3'-quinolone, was determined. Overall, 97% of Enterobacteriaceae and 100% of staphylococci were inhibited by < or = 8.0 mg/L of RO 23-9424. Only 60% of Pseudomonas aeruginosa and 80-90% of Pseudomonas spp. and Xanthomonas maltophilia were inhibited by this concentration. Enterococci and Listeria monocytogenes were resistant to RO 23-9424. Clinical isolates of Moraxella catarrhalis, Streptococcus spp., and Corynebacterium jekeium were all susceptible to < or = 8.0 mg/L of RO 23-9424. This drug's antimicrobial activity was superior to that of its two components fleroxacin and desacetylcefotaxime against the organisms tested. Using < or = 8.0 mg/L and > or = 32 mg/L respectively as the susceptible and resistant MIC breakpoints for RO 23-9424, the regression analysis-derived disc diffusion zone diameter breakpoints for the 30 micrograms disc are: susceptible > or = 19 mm, intermediate 16-18 mm, and resistant < or = 15 mm.