Beta-lactam susceptibilities and prevalence of ESBL-producing isolates among more than 5000 European Enterobacteriaceae isolates.

In vitro susceptibility to 15 beta-lactam antibiotics was evaluated using Enterobacteriaceae isolated during the SENTRY Antimicrobial Surveillance Program. Piperacillin/tazobactam was the most active penicillin against Escherichia coli, Proteus mirabilis, Klebsiella oxytoca and Klebsiella pneumoniae (94.9%, 98.3%, 87.4% and 82.9% of isolates susceptible). Of the cephalosporins, cefepime was most effective against Escherichia coli, Proteus mirabilis and Enterobacter cloacae (99.2%, 96.3% and 95.2% of isolates susceptible, respectively) and cefoxitin against Klebsiella oxytoca and Klebsiella pneumoniae (98.6% and 95.6% of isolates susceptible). Carbapenems had excellent activity (> or =99.5% of all isolates). ESBL-production was confirmed with the ESBL-Etest and disk diffusion test in 1.3% of Escherichia coli isolates, 18.4% of Klebsiella pneumoniae, 12.6% of Klebsiella oxytoca and 5.3% of Proteus mirabilis isolates.

Resistance integrons and super-integrons.

Integrons are genetic elements composed of a gene encoding an integrase, gene cassettes and an integration site for the gene cassettes (att). The integrase excises and integrates the gene cassettes from and into the integron, but integrons themselves are not mobile. Two groups of integrons are known: resistance integrons and super-integrons. Nearly all known gene cassettes from resistance integrons encode resistance to antibiotics or disinfectants. These integrons are found on transposons, plasmids and the bacterial chromosome. Gene cassettes in super-integrons encode a variety of different functions. Super-integrons are located on the bacterial chromosome. More than 100 gene cassettes may be present, in contrast to resistance integrons where less than ten cassettes are present. Many species harbour super-integrons, which are species-specific, whereas particular resistance integrons can be found in a variety of species. The gene cassettes in resistance integrons probably originated from super-integrons. In the last few years, a variety of new gene cassettes have been described. Many of these encode resistance against newer antibiotics such as cephalosporins and carbapenems. Resistance integrons have been found in isolates from a wide variety of sources, including food.

[Modern diagnostics for urological infections]. / Moderne Diagnostik urologischer Infektionen.

This paper provides a short overview of modern, molecular-based diagnostic procedures of urogenital tract infections. Although gaining importance, molecular methods have not yet become a reliable substitution for the classic procedures in terms of costs and quality standards. As an example of a new molecular approach in microbiology, a method for the detection of the most relevant uropathogens in a single PCR is presented. Furthermore, the development of a real time PCR is described.

Mutations in GyrA, ParC, MexR and NfxB in clinical isolates of Pseudomonas aeruginosa.

The target enzymes GyrA and ParC and two efflux pump regulatory genes mexR and nfxB were analysed to determine changes associated with fluoroquinolone resistance in Pseudomonas aeruginosa. Both low- and high-level ciprofloxacin resistance was associated with a Thr-83Ile substitution in GyrA. A ParC Ser-80Leu substitution was found in highly resistant isolates in tandem with the Thr-83Ile substitution in GyrA. Mutations in the efflux regulatory genes were associated with resistance only when in tandem with a mutation in GyrA or ParC. These data show that the main mechanism of fluoroquinolone resistance in P. aeruginosa is mediated primarily through mutations in GyrA, and that mutations in ParC and the efflux regulatory genes are secondary.

In vitro antibacterial activity and pharmacodynamics of new quinolones.

This synopsis of published literature summarises data on the in vitro antibacterial activity and pharmacodynamics of fluoroquinolones. Data were compiled for ciprofloxacin, levofloxcin, moxifloxacin, gatifloxacin, grepafloxacin, gemifloxacin, trovafloxacin, sitafloxacin and garenoxacin. All of these quinolones are almost equipotent against gram-negative bacteria but demonstrate improved activity against gram-positive species. The new quinolones are uniformly active against gram-positive species except Streptococcus pneumoniae; against which gemifloxacin, sitafloxacin and garenoxacin are one to two dilution steps more active than moxifloxacin. All of the new quinolones except gemifloxacin demonstrate enhanced activity against anaerobes. Since all the new quinolones show similar activity against the major respiratory tract pathogens except Streptococcus pneumoniae and members of the family Enterobacteriaceae, their pharmacokinetics and pharmacodynamics will be clinically relevant differentiators and determinants of their overall activity and efficacy. In vitro simulations of serum concentrations revealed that (i). gemifloxacin and levofloxacin were significantly and gatifloxacin moderately less active than moxifloxacin against Streptococcus pneumoniae and Staphylococcus aureus, and (ii). resistant subpopulations emerged following exposure to levofloxacin and gatifloxacin (gemifloxacin not yet published) but not to moxifloxacin. The emergence of resistance is a function of drug concentrations achievable in vivo and the susceptibility pattern of the target organisms. Therefore, the use of less potent fluoroquinolones with borderline or even suboptimal pharmacokinetic/pharmacodynamic surrogate parameters will inadvertently foster the development of class resistance. Drugs with the most favourable pharmacokinetic/pharmacodynamic characteristics should be used as first-line agents in order to preserve the potential of this drug class and, most importantly, to provide the patient with an optimally effective regimen.

Fluoroquinolones: structure and target sites.

The quinolones are a potent group of drugs that target the essential bacterial enzymes DNA gyrase and topoisomerase IV. DNA gyrase is the primary target of Gram negative organisms however, it is topoisomerase IV that is the primary target of Gram positive organisms. Within these enzymes is a highly conserved region centered round the active site where resistance mutations occur. These mutations are almost always identical, irrespective of organism. In spite of the homology of this region, amino acid sequence analysis shows that there are defined differences between the Gram groups, particularly in topoisomerase IV, and it is speculated that herein lies the origin of target preference. Since the first quinolone nalidixic acid was developed, the quinolones have undergone structural modifications, in particular the addition of a fluorine at position 6, to produce the fluoroquinolones. This has seen their potency and pharmakokinetic profile greatly increase. In vitro selection of resistance mutations has allowed the observation of how resistance is acquired and some of the modifications in newer fluoroquinolones have resulted in the shift of primary target from topoisomerase IV to gyrase with Gram positives. Curiously, purified topoisomerase IV is still more sensitive even if gyrase is the primary target. Gyrase remains the primary target for Gram negatives.

Activities of the glycylcycline tigecycline (GAR-936) against 1,924 recent European clinical bacterial isolates.

The in vitro activities of tigecycline against 1,924 clinical isolates were examined. The new glycylcycline exhibited excellent activity against all gram-positive cocci (MICs at which 90% of the isolates tested were inhibited [MIC(90)s],

mecA gene is widely disseminated in Staphylococcus aureus population.

Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most important causes of hospital infections worldwide. High-level resistance to methicillin is caused by the mecA gene, which encodes an alternative penicillin-binding protein, PBP 2a. To determine the clonal relationships between methicillin-susceptible S. aureus (MSSA) and MRSA, we typed 1,069 S. aureus isolates (493 MSSA isolates and 576 MRSA isolates), collected mainly in North American and European hospitals between the 1960s and the year 2000, using pulsed-field gel electrophoresis and ribotyping. Of 10 widespread S. aureus lineages recognized, 8 had corresponding mecA-positive strains. Multiresistant MRSA strains are found in hospitals worldwide, while unrelated and more susceptible strains represent less than 1% of the MRSA population. This supports the hypothesis that horizontal transfer plays an important role in the dissemination of the mecA gene in the S. aureus population.

Activity of quinolones against gram-positive cocci: mechanisms of drug action and bacterial resistance.

The quinolones are a potent class of antimicrobial agents that target two essential enzymes of bacterial cells: DNA gyrase and topoisomerase IV. Resistance is mediated chiefly through stepwise mutations in the genes that encode these enzymes, leading to alterations of the target site. These mutations occur in an area called the "quinolone resistance determining region". In gram-positive organisms, mutations occur more often in topoisomerase IV than in DNA gyrase. This target preference appears to depend upon two factors: the species of organism and the selecting drug. Resistance can be enhanced by a decrease in intracellular drug concentration, which is mediated through efflux pumps. The newer generation of fluoroquinolones and non-fluorinated quinolones exhibits enhanced activity against gram-positive organisms compared to the older members of this drug class, although development of resistance to these drugs has been demonstrated in vitro. This review gives a chronological perspective of the literature on the action of DNA gyrase and topoisomerase IV and the mechanisms of resistance to quinolones in staphylococci, streptococci and enterococci.

In vitro activity of AZD2563, a novel oxazolidinone, against European Gram-positive cocci.

AZD2563 is a new oxazolidinone that has targeted activity against Gram-positive bacteria. The in vitro activity of AZD2563 and nine comparators against 1543 European enterococcal, staphylococcal and streptococcal isolates was determined. The compound is a potent oxazolidinone, with no isolate tested displaying an MIC > 4 mg/L and 94.4% having an MIC < or =2 mg/L. Compared with linezolid, the distribution of MICs for all species of bacteria tested, with the exception of Enterococcus faecium, was shifted by one or two dilution steps to lower values for AZD2563. Thus, this oxazolidinone is a promising new antibiotic for the treatment of infections with Gram-positive cocci.

Therapeutic options in an era of decreasing antimicrobial susceptibility.

Antimicrobial resistance among human pathogens has evolved to the multidrug-resistant phase. Consequently, preserving antimicrobial efficacy is now an important clinical consideration when selecting an agent for therapy. A new, in vitro measure of potency called the mutant prevention concentration (MPC) allows clinicians to select antibacterials on the basis of an agent's ability to restrict the selection of resistant mutants. With some pathogens, C-8-methoxy fluoroquinolones are expected to be quite effective at restricting the selection of resistance because the value of MPC is below the human serum drug concentration achieved with standard doses. Clinical trials are now needed to determine how well preclinical profiles of these drugs correlate with the prevention of resistance.

Three cases of bacterial meningitis after spinal and epidural anesthesia.

During a 3-year period, three cases of bacterial meningitis developing after spinal or epidural anesthesia were observed at one hospital in Germany. The causative organisms were Streptococcus salivarius (2 cases) and Staphylococcus aureus (1 case). In the first two cases, contamination of the needle by oropharyngeal flora of the anesthesiological team was likely but remained unproven. In the third case, a nasal swab obtained from the operating anesthesiologist yielded a Staphylococcus aureus strain whose genotypic profile was identical to that of the patient's strain. Infection control procedures for spinal anesthesia are discussed.