Joan Fung-Tomc

Activity of quinolones in the Ames Salmonella TA102 mutagenicity test and other bacterial genotoxicity assays.

Eight quinolones were examined for their bacterial mutagenicity in the Ames Salmonella TA102 assay and for their effects in other bacterial genotoxicity assays. In the quantitative Ames plate incorporation assay, all eight quinolones induced His+ deletion reversion in Salmonella tester strain TA102, with maximum reversion observed at about two to eight times the MIC. The quinolones also induced the SOS response. At quinolone concentrations close to the MIC, SOS cell filamentation gene sulA was induced in sulA::lacZ fusion strain Escherichia coli PQ37. RecA-mediated cleavage of lambda repressor in lambda::lacZ fusion strain E. coli BR513 was measurable at about 10 times the MIC, though no induction occurred at 20 micrograms of nalidixic or oxolinic acid per ml. Genotoxicity of quinolones also was observed in the Bacillus subtilis DNA repair assay, in which the mutant strain M45 (recA) was more susceptible to quinolones than its parent strain, H17 (rec+). The results from these analyses indicate that quinolones induce SOS functions and are mutagenic in bacteria; these properties correspond to their antimicrobial activities. Images

Activity of cefepime against ceftazidime- and cefotaxime-resistant gram-negative bacteria and its relationship to beta-lactamase levels.

One hundred clinical isolates resistant to ceftazidime and/or cefotaxime were examined for susceptibility to cefepime. The most frequently encountered ceftazidime-cefotaxime-resistant strains belonged to the genera Enterobacter, Pseudomonas, and Citrobacter. Among these strains, 92% were resistant to cefoperazone, 91% were resistant to cefotaxime, 84% were resistant to ceftazidime, and 6% were resistant to cefepime. Of the members of the family Enterobacteriaceae, 57% were resistant to ceftriaxone. The six strains resistant to cefepime were all Pseudomonas aeruginosa and were resistant to both cefotaxime and ceftazidime. Cefepime-resistant P. aeruginosa strains had exceptionally high levels of beta-lactamase activity, higher than the levels found in strains resistant to ceftazidime but susceptible to cefepime. The beta-lactamases from the cefepime-resistant strains were type I (Richmond-Sykes), were constitutively produced, and did not have increased affinity or hydrolytic activity for cefepime. Thus, cefepime was active against most gram-negative bacteria which have developed resistance to the broad-spectrum cephalosporins, and resistance to cefepime in P. aeruginosa appears to be associated with higher beta-lactamase levels than in cefepime-susceptible strains.

Antimicrobial Evaluation of Nocathiacins, a Thiazole Peptide Class of Antibiotics

ABSTRACT Nocathiacins are cyclic thiazolyl peptides with inhibitory activity against gram-positive bacteria. BMS-249524 (nocathiacin I), identified from screening a library of compounds against a multiply antibiotic-resistant Enterococcus faecium strain, was used as a lead chemotype to obtain additional structurally related compounds. The MIC assay results of BMS-249524 and two more water-soluble derivatives, BMS-411886 and BMS-461996, revealed potent in vitro activities against a variety of gram-positive pathogens including methicillin-resistant Staphylococcus aureus, penicillin-resistant Streptococcus pneumoniae, vancomycin intermediate-resistant S. aureus, vancomycin-resistant enterococci, Mycobacterium tuberculosis and Mycobacterium avium. Analysis of killing kinetics revealed that these compounds are bactericidal for S. aureus with at least a 3-log10 reduction of bacterial growth within 6 h of exposure to four times the MICs. Nocathiacin-resistant mutants were characterized by DNA sequence analyses. The mutations mapped to the rplK gene encoding the L11 ribosomal protein in the 50S subunit in a region previously shown to be involved in the binding of related thiazolyl peptide antibiotics. These compounds demonstrated potential for further development as a new class of antibacterial agents with activity against key antibiotic-resistant gram-positive bacterial pathogens.

Ciprofloxacin-induced, low-level resistance to structurally unrelated antibiotics in Pseudomonas aeruginosa and methicillin-resistant Staphylococcus aureus.

The effects of ciprofloxacin on the rates of development of low-level resistance to other antibiotics were determined in vitro. Three methicillin-resistant Staphylococcus aureus and two Pseudomonas aeruginosa clinical strains were grown overnight in Mueller-Hinton broth with or without subinhibitory concentrations (1/2, 1/4, and 1/8 MICs) of ciprofloxacin or an aminoglycoside and then quantitatively plated onto medium containing 4 or 8 times the MICs of various antibiotics. The spontaneous mutational frequencies were determined and compared with those of cells not exposed to ciprofloxacin. Exposure of methicillin-resistant S. aureus strains to ciprofloxacin resulted in a > 100-fold increase in the isolation of variants with decreased susceptibilities to ciprofloxacin, tetracycline, imipenem, fusidic acid, and gentamicin, but not vancomycin. Likewise, a > 100-fold increase in the isolation of variants with decreased susceptibilities to ciprofloxacin and imipenem (35-fold) in P. aeruginosa A21213 was observed, and a > 100-fold increase in the isolation of variants with decreased susceptibilities to ciprofloxacin, amikacin, and cefepime in P. aeruginosa A22379 was observed. On the other hand, exposure of these strains to an aminoglycoside did not influence the development of resistance to nonaminoglycoside drugs. These results indicate that exposure to subinhibitory levels of ciprofloxacin can promote the development of low-level resistance to antibiotics with different modes of action.

Correlation between Genotype and Phenotypic Categorization of Staphylococci Based on Methicillin Susceptibility and Resistance

ABSTRACT Positive correlation between methicillin and oxacillin susceptibility test results and the detection of themecA gene was observed forStaphylococcusaureus, S. epidermidis, and S. haemolyticus as well as among mecA+ strains of other species of coagulase-negative staphylococci (CNS). However, at least 50% of themecA-negative strains of these other species of CNS were falsely classified as methicillin and oxacillin resistant.

In Vitro and In Vivo Activities of a Novel Cephalosporin, BMS-247243, against Methicillin-Resistant and -Susceptible Staphylococci

ABSTRACT The recent emergence of methicillin-resistant Staphylococcus aureus (MRSA) with decreased susceptibility to vancomycin has intensified the search for alternative therapies for the treatment of infections caused by this organism. One approach has been to identify a β-lactam with improved affinity for PBP 2a, the target enzyme responsible for methicillin resistance in staphylococci. BMS-247243 is such a candidate, with MICs that inhibit 90% of isolates tested (MIC90s) of 4, 2, and 8 μg/ml for methicillin-resistant strains of S. aureus, S. epidermidis, and S. haemolyticus, respectively, as determined on plates with Mueller-Hinton agar and 2% NaCl. The BMS-247243 MICs for MRSA were minimally affected by the susceptibility testing conditions (inoculum size, prolonged incubation, addition of salt to the test medium) or by staphylococcal β-lactamases. BMS-247243 MIC90s for methicillin-susceptible staphylococcal species ranged from ≤0.25 to 1 μg/ml. The BMS-247243 MIC90 for β-lactamase-producing S. aureus strains was fourfold higher than that for β-lactamase-nonproducing strains. BMS-247243 is hydrolyzed by staphylococccal β-lactamases at 4.5 to 26.2% of the rates measured for cephaloridine. The affinity of BMS-247243 for PBP 2a was >100-fold better than that of methicillin or cefotaxime. BMS-247243 is bactericidal for MRSA, killing the bacteria twice as fast as vancomycin. These in vitro activities of BMS-247243 correlated with its in vivo efficacy against infections in animals, including the neutropenic murine thigh and rabbit endocarditis models involving MRSA strains. In conclusion, BMS-247243 has in vitro and in vivo activities against methicillin-resistant staphylococci and thus may prove to be useful in the treatment of infections caused by these multidrug-resistant organisms.

In vitro antifungal and fungicidal spectra of a new pradimicin derivative, BMS-181184.

A new pradimicin derivative, BMS-181184, was compared with amphotericin B and fluconazole against 249 strains from 35 fungal species to determine its antifungal spectrum. Antifungal testing was performed by the broth macrodilution reference method recommended by the National Committee for Clinical Laboratory Standards (document M27-P, 1992). BMS-181184 MICs for 97% of the 167 strains of Candida spp., Cryptococcus neoformans, Torulopsis glabrata, and Rhodotorula spp. tested were < or = 8 micrograms/ml, with a majority of MICs being 2 to 8 micrograms/ml. Similarly, for Aspergillus fumigatus and 89% of the 26 dermatophytes tested BMS-181184 MICs were < or = 8 micrograms/ml. BMS-181184 was fungicidal for the yeasts, dermatophytes, and most strains of A. fumigatus, although the reduction in cell counts was less for A. fumigatus than for the yeasts. BMS-181184 was active against Sporothrix schenckii, dematiaceous fungi, and some members of the non-Aspergillus hyaline hyphomycetes. BMS-181184, however, was not fungicidal against members of the family Dematiaceae. BMS-181184 lacked activity or had poorer activity (MICs, > or = 16 micrograms/ml) against Aspergillus niger, Aspergillus flavus, Malassezia furfur, Fusarium spp., Pseudallescheria boydii, Alternaria spp., Curvularia spp., Exserohilum mcginnisii, and the zygomycetes than against yeasts. The activity of BMS-181184 was minimally (twofold or less) affected by changes in testing conditions (pH, inoculum size, temperature, the presence of serum), testing methods (agar versus broth macrodilution), or test media (RPMI 1640, yeast morphology agar, high resolution test medium). Overall, our results indicate that BMS-181184 has a broad antifungal spectrum and that it is fungicidal to yeasts and, to a lesser extent, to filamentous fungi.

Antibacterial activities of cefprozil compared with those of 13 oral cephems and 3 macrolides.

Thirteen oral cephems (cefprozil, loracarbef, cefaclor, cefuroxime axetil, cefpodoxime proxetil, cefetamet pivoxil, cefixime, cefdinir, cefadroxil, cephradine, cephalexin, cefatrizine, and cefroxadine), the cephalosporin class representative cephalothin, cefazolin, and the macrolides erythromycin, clarithromycin, and azithromycin were compared for their antibacterial activities against 790 recent clinical isolates. These oral agents differed in their spectra and antibacterial potencies against community-acquired pathogens.

Susceptibility of bacterial isolates to β-lactam antibiotics from U.S. clinical trials over a 5-year period

Results are reported for agar dilution susceptibility testing of 3,075 isolates of aerobic bacteria collected from >200 U.S. institutions, located in 30 different states. These isolates were collected from 1987 through 1991 from patients who participated in cefepime clinical trials. Cefepime susceptibility was compared with ceftazidime, cefotaxime, ceftriaxone, cefoperazone, and imipenem. To avoid duplication of strains, only initial isolates were included. Cefepime minimum inhibitory concentration (MIC90) values for Enterobacteriaceae were < or = 0.5 microg/mL, except for two species, Citrobacter freundii and Providencia stuartii, with MIC90 values of 2 and 1, respectively. The MIC90 values of the other cephalosporins were higher, especially for Enterobacter aerogenes and C. freundii. The MIC90 values of cefepime for methicillin-susceptible Staphylococcus aureus (4 microg/mL) and Pseudomonas aeruginosa (8 microg/mL) were similar to those of cefotaxime for S. aureus (4 microg/mL), and to ceftazidime for P. aeruginosa (8 microg/mL). Streptococcus pneumoniae was similar in susceptibility to cefotaxime at 0.06 microg/mL. The activity of cefepime against a diverse group of gram-positive and gram-negative (1987-1991) bacteria isolates demonstrates the excellent activity of cefepime compared to third-generation cephalosporins and imipenem, particularly among C. freundii and E. aerogenes isolates, which were often resistant to other cephalosporins.

Frequency of in vitro resistance of Pseudomonas aeruginosa to cefepime, ceftazidime, and cefotaxime.

The selection frequencies of cefepime (BMY 28142), ceftazidime, and cefotaxime resistance among Pseudomonas aeruginosa strains were determined. Cefepime-resistant mutants were not selected by cefepime (frequency, less than 10(-11)). Ceftazidime- and cefotaxime-resistant mutants were isolated at frequencies of 10(-5) to 10(-10) and were often cross-resistant. However, cefepime resistance among ceftazidime- and cefotaxime-resistant mutants was rare. Selected mutants resistant to cefepime constitutively produced 40- to 450-fold more beta-lactamase than did the parent strain.