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National survey of susceptibility to antimicrobials amongst clinical isolates of Pseudomonas aeruginosa.

Between September and December 1993, each of 24 hospitals in the UK collected up to 100 consecutive clinical isolates of Pseudomonas aeruginosa and sent these to the London Hospital Medical College (LHMC). Of 2184 cultures received, 1991 contained viable P. aeruginosa. Minimum inhibitory concentrations (MICs) of antimicrobials were determined by agar dilution. The frequencies of resistance to low breakpoints were as follows: gentamicin, MIC > 2 mg/L, 11.7%; amikacin, MIC > 4 mg/L, 10.5%, carbenicillin, MIC > 128 mg/L, 11.7%; azlocillin, MIC> 16 mg/L, 10.9%; ceftazidime, MIC > 4 mg/L, 9.6%; ciprofloxacin, MIC > 1 mg/L, 8.1%; imipenem, MIC > 4 mg/L 2.5% and meropenem, MIC > 4 mg/L, 1.1%. Resistance to each antimicrobial except amikacin was commoner among the 134 isolates from patients in intensive care units (ICUs) than amongst the 1042 isolates from other in-patients (P < 0.01). Resistance to penicillins and ceftazidime, though not to other agents, was rarer among the 797 isolates from out-patients than amongst those from non-ICU in-patient (P < 0.01). Compared to a similar study in 1982, during which 1866 isolates had been examined, the frequency of resistance to the aminoglycosides increased (P < 0.05) as had those to the penicillins and ceftazidime (P < 0.01). Ciprofloxacin and the carbapenems were not tested in 1982. Cross-resistance patterns suggested that the increases in resistance to aminoglycosides and beta-lactams were largely a reflection of greater numbers of isolates with barrier or efflux mechanisms and were not due to an increase in isolates with antibiotic-degrading enzymes. The participating hospitals mostly employed Stokes' disc diffusion method and, when the results were compared to the MICs determined at the LHMC, fewer than 9% of the isolates reported as susceptible were found to be resistant. However, up to 72% of those reported by the hospitals as resistant were found to be susceptible.

Anti-Bacterial Agents↗

Bactericidal effect of beta-lactams and amikacin alone or in association against Klebsiella pneumoniae producing extended spectrum beta-lactamase.

Ten extended spectrum beta-lactamases producing strains of Klebsiella pneumoniae characterized by analytical isoelectric focusing and studied for their susceptibility to beta-lactam antibiotics, either alone or in combination with a beta-lactamase inhibitor (clavulanic acid and sulbactam) and in association with amikacin. The extended spectrum beta-lactamases were derived from either TEM (CTX-1 = TEM-3) or SHV (CAZ-4 = SHV-5). Killing curves were studied with antibiotics at clinical by achievable concentrations, at MIC and MIC x 4. At MIC, cefotetan, cefotaxime and ceftazidime lacked bactericidal activity. Imipenem was more rapidly bactericidal than meropenem or co-amoxiclav. At MIC x 4, cefotetan and cefotaxime exhibited bactericidal effect but this was less than for imipenem which gave a reduction of 4 log10 of the inoculum. Cefotaxime plus sulbactam gave no bactericidal effect compared with cefotaxime plus co-amoxiclav. A bactericidal effect with cefotaxime plus sulbactam was seen with the addition of amikacin. At clinical concentrations cefotaxime plus co-amoxiclav +/- amikacin was as efficient as imipenem +/- amikacin with a rapid bactericidal effect (5-6 log10 in 30-60 min). We proposed that cefotaxime+co-amoxiclav might be considered as an alternative to imipenem for the treatment of extended spectrum beta-lactamase associated K. pneumoniae injections.

Amikacin↗

The ability of beta-lactam antibiotics to select mutants with derepressed beta-lactamase synthesis from Citrobacter freundii.

The ability of beta-lactam compounds to induce synthesis of the class C beta-lactamase of Citrobacter freundii was assessed both directly and indirectly, the latter by measurement of the different susceptibilities of strains with inducible and depressed beta-lactamase synthesis and those of a beta-lactamase-negative strain. Most beta-lactam compounds were poor inducers but ampicillin, amoxycillin and cephalexin were moderately good inducers and cefoxitin, imipenem and meropenem were strong inducers. The ability of the compounds to select mutants in which the synthesis of the beta-lactamase was derepressed was also assessed. Imipenem and temocillin, which had a high degree of stability to the beta-lactamase failed to select such mutants or were very poor selectors. In contrast, most other compounds showed considerable lability to the beta-lactamase and readily selected derepressed mutants, whether they were strong inducers of beta-lactamase synthesis (for example cefoxitin) or poor inducers (cefotaxime, ceftazidime and many other compounds). Thus it appears that lability to the beta-lactamase is a more important factor in determining whether or not a compound will select derepressed mutants than the power of the compound as an inducer of beta-lactamase synthesis, since induction results in the production of less beta-lactamase than derepression as a result of mutation.

Anti-Bacterial Agents↗

Susceptibility of Streptococcus pneumoniae strains isolated in Italy to penicillin and ten other antibiotics.

The susceptibility of 312 Streptococcus pneumoniae strains isolated during 1993 in Italy has been studied. Overall resistance to penicillin, erythromycin and co-trimoxazole was 5.5, 6 and 18% respectively. The penicillin resistant isolates (nine displaying low- and eight high-level resistance) were further tested against cefotaxime, ceftriaxone, cefepime, cefodizime, ceftazidime, imipenem, meropenem and rifampicin. The low-level penicillin resistant strains were inhibited by all these drugs. Among the eight high-level penicillin resistant pneumococci, four were resistant to one or more cephalosporins. Imipenem and rifampicin remained effective against all isolates. In comparison with other European Mediterranean countries, penicillin resistance is a minor threat at present in Italy.

Anti-Bacterial Agents↗

Activity of biapenem (LJC 10627) against 51 imipenem-resistant bacteria and selection and characterisation of biapenem-resistant mutants.

For wild-type bacteria the activity of biapenem was similar to that of imipenem, but for 51 imipenem-resistant strains meropenem was more active than either. When penicillin-binding protein 2a (PBP 2a) was expressed in Staphylococcus aureus biapenem had reduced activity. Mutant bacteria with decreased susceptibility to biapenem were selected in agar. Most of the mutant Gram negative bacteria were unstable and readily reverted to susceptible. The mutant Proteus vulgaris and Pseudomonas aeruginosa lacked an outer membrane protein. Biapenem-resistant S. aureus could be selected only from MRSA.

Drug Resistance, Microbial↗

In-vitro activity of cephalosporins alone and combined with sulbactam against various strains of Acinetobacter baumannii with different antibiotic resistance profiles.

The beta-lactamases of six strains of Acinetobacter baumannii were investigated using analytical isoelectric focusing, and the MIC values for 16 beta-lactam antibiotics determined against the strains. Three strains produced a chromosomal cephalosporinase (pI > 8.5), one strain a CARB-5 beta-lactamase (pI = 6.35), one strain a TEM-1 penicillinase (pI = 5.4) as well as a cephalosporinase (pI > 8.5), and one carbapenem-resistant strain a beta-lactamase with a pI > 8.5. Ceftazidime, cefepime, cefpirome, imipenem and meropenem were found to be the most effective beta-lactams against five strains. Sulbactam was active against four of the strains at 1-4 mg/L and showed enhanced killing effects with cefpirome.

Acinetobacter↗

Carbapenem resistance in Pseudomonas aeruginosa from cystic fibrosis patients.

The evolution of imipenem resistance was evaluated in Pseudomonas aeruginosa sequentially isolated from 42 patients with cystic fibrosis. Susceptibility was determined using a commercial microdilution system and imipenem resistance was confirmed by the agar dilution technique. Resistance to imipenem increased during the years from 1988 to 1992. A total of 12 patients (28.5%) carried resistant strains (11.6% of the total P. aeruginosa isolates) but only two of them were treated with the carbapenem. The other patient under imipenem treatment did not harbour resistant isolates. Sixty-four per cent of the imipenem resistant isolates were also meropenem resistant and showed low susceptibility to the other beta-lactams and tobramycin and amikacin. Twenty-one strains were selected for biochemical study. Imipenem susceptible strains showed normal OprD in two strains and diminished OprD in two more. Five strains with MIC of imipenem of 4-8 mg/L lacked OprD while another two had a band with decreased density. All strains with MIC higher than 8 completely lacked this band in western-blot analysis. Imipenem MICs of 0.5-2 mg/L only slightly increased to 1-4 mg/L when a pattern of beta-lactamase derepression was observed. While those with imipenem MICs between 8-16 mg/L increased the imipenem MIC to 16-64 mg/L in the population with a beta-lactamase derepression phenotype.

Adolescent↗

Evaluation of high-level carbapenem resistance in atypical Serratia marcescens by a comparison with its revertants.

A clinical isolate of Serratia marcescens (FHSM4055) was highly resistant to carbapenem (MIC of imipenem, 100 mg/L) and was atypical in that it was negative in the aesculin-hydrolysis test. This parent strain was compared with the revertants S1 and S5, which had lost resistance to imipenem alone and to imipenem and piperacillin respectively. The MICs of imipenem for the revertants were 16- to 32-fold lower than that for the parent strain. Only the crude extract of the parent strain had imipenem-hydrolyzing activity (0.27 unit/mg: panipenem-, meropenem- and biapenem-hydrolyzing activities were 53.4, 22.6 and 26.7% of the imipenem-hydrolyzing activity, respectively). From the isoelectric focusing profiles of beta-lactamases detected with nitrocefin, the parent strain produced beta-lactamases with pls of 9.2, 8.7 and 5.5, whereas the revertants did not produce the pI 8.7 beta-lactamase, which is a metalloenzyme. In the induction test, the pI 8.7 beta-lactamase was constitutive. These results indicated that the constitutive pI 8.7 beta-lactamase contributes to the carbapenem resistance of the parent strain. On the other hand, the MICs of imipenem for the revertants were 8- to 16-fold higher than that for the typical S. marcescens reference strain IFO3736, which was susceptible to various beta-lactam antibiotics other than first generation cephalosporins (according to the disk sensitivity test). This result revealed that the revertants were low-level resistant to carbapenem. The protein corresponding to the 42 kDa porin of strain IFO3736 (the major transport-channel) was absent on the outer membrane protein profiles of the parent strain and the revertants, and [3H]glucose uptake concomitantly decreased. Hence a combination of constitutive pI 8.7 metallo-beta-lactamase and decreased outer membrane permeability are major factors involved in the high-level carbapenem resistance of S. marcescens.

Carbapenems↗

Comparative in-vitro activities of quinupristin-dalfopristin against Gram-positive bloodstream isolates.

The in-vitro activity of quinupristin-dalfopristin was compared with those of vancomycin, teicoplanin, erythromycin, clarithromycin, rifampicin, imipenem, meropenem, ciprofloxacin and sparfloxacin against 414 bloodstream isolates of Gram-positive cocci. Quinupristin-dalfopristin inhibited strains of Streptococcus pyogenes and Streptococcus agalactiae at 0.12 mg/L, methicillin- and/or erythromycin-resistant Staphylococcus aureus and Staphylococcus epidermidis at 0.5 mg/L, Staphylococcus haemolyticus, Enterococcus faecium, Streptococcus pneumoniae, Streptococcus mitis, Streptococcus bovis, Streptococcus sanguis and Streptococcus anginosus at 1 mg/L and Enterococcus faecalis at 8 mg/L.

Anti-Bacterial Agents↗

Correlation between genotype and beta-lactamases of clinical and environmental strains of Stenotrophomonas maltophilia.

Heterogeneity of beta-lactamase production by 17 clinical and nine environmental isolates of Stenotrophomonas maltophilia was investigated using MICs of six different beta-lactam antibiotics, isoelectric focusing (IEF) and pulsed field gel electrophoresis. There was no clear correlation between the results of IEF, genotype and MIC determination. Environmental isolates were more susceptible than clinical isolates; eight clinical and none of the environmental isolates expressed high-level resistance to meropenem. Only two isolates expressed high-level resistance to ceftazidime. These results indicate that further studies are required to elucidate the extent of genetic heterogeneity within the L1 and L2 beta-lactamase genes.

Anti-Bacterial Agents↗

Antipneumococcal activity of MEN 10700, a new penem, compared with other compounds, by MIC and time-kill kinetics.

The antipneumococcal activity of MEN 10700 was compared with those of nine other compounds by MIC and time-kill kinetics. MIC90s (mg/L) of 202 penicillin-susceptible, -intermediate and -resistant pneumococci were: 0.06, 1.0 and 2.0 (MEN 10700); 0.06, 0.5-1.0 and 2.0 (amoxycillin +/- clavulanate); 0.06, 0.5 and 4.0 (cefotaxime); 0.125, 0.5 and 2.0 (cefepime); 0.016, 0.125 and 0.25 (imipenem); 0.03, 0.5 and 1.0 (meropenem); 2.0 (ciprofloxacin); 0.125, >64.0 and >64.0 (clarithromycin); and 0.5 (vancomycin). Time-kill kinetics showed that MEN 10700, at 4 x MIC, was bactericidal for all 12 isolates tested at 4 x MIC. Kinetics of other beta-lactams were similar to those of MEN 10700, relative to MICs. Ciprofloxacin, at 4 x MIC, was uniformly bactericidal after 24 h. Clarithromycin exhibited slow kill kinetics, after 24 h. Vancomycin was bactericidal against 11/12 isolates at 2 x MIC after 24 h.

Anti-Bacterial Agents↗

High prevalence of resistance to clindamycin in Bacteroides fragilis group isolates.

Susceptibility to anti-anaerobic agents in the Bacteroides fragilis group varies according to the geographical region studied. In recent years there has been a reduction in the susceptibility of such isolates, particularly to antibiotics such as clindamycin and cefoxitin. The antimicrobial susceptibilities of 100 isolates of the B. fragilis group isolated in 1998 from faecal samples of healthy people to clindamycin and five other anti-anaerobic agents were determined. Meropenem, metronidazole and trovafloxacin showed excellent activity against all isolates. The efficacy of cefoxitin was low, with only 46% of isolates susceptible. A high prevalence of resistance to clindamycin (49% of isolates) was observed.

Adolescent↗

beta-lactamase expression in Plesiomonas shigelloides.

We have examined the production of beta-lactamases by 11 clinical and nine environmental isolates of Plesiomonas shigelloides from Czechoslovakia, the Czech Republic and Cuba. Ten isolates (50%) expressed detectable amounts of a chromosomally encoded, non-inducible beta-lactamase, though all isolates showed a broadly similar resistance profile: low-level resistance to ampicillin and higher-level resistance to carbenicillin. All strains were susceptible to cephalosporins and meropenem. Three clinical isolates expressed a beta-lactamase similar to a class 2c carbenicillinase, with a pI of 5.2 and three expressed an enzyme similar to a class 2d oxacillinase, with a pI of 5.3. The environmental isolates produced a variety of penicillinases, indicating that there is a reservoir of heterogeneous beta-lactamase genes in this species.

Anti-Bacterial Agents↗

Expression of the carbapenemase gene (cfiA) in Bacteroides fragilis.

Bacteroides fragilis strains were studied to examine carbapenemase gene (cfiA) expression and insertion sequence (IS) element promoters. High-level resistance was associated with high meropenemase activity and IS elements upstream of cfiA. Sequencing revealed two element types; IS1187 and elements related to IS942 and IS1170. The latter was implicated in the conversion to carbapenem resistance in a cfiA-positive isolate during imipenem therapy. Two strains showing low-level resistance, and strains susceptible to meropenem, did not possess IS elements upstream of cfiA. The prevalence in Nottingham of clinical isolates of B. fragilis with cfiA and efficient IS element promoters was low (0.6%).

Bacterial Proteins↗

In vitro activity of garlic oil and four diallyl sulphides against antibiotic-resistant Pseudomonas aeruginosa and Klebsiella pneumoniae.

The in vitro antibacterial activities of garlic oil and four diallyl sulphides naturally occurring in this oil were studied against Pseudomonas aeruginosa and Klebsiella pneumoniae (total 237 clinical isolates). Garlic oil at 4 x MIC could reduce original inoculum to diallyl disulphide > diallyl trisulphide (DAT) > diallyl tetrasulphide (DATS) (P < 0.05). Most interactions of ceftazidime, gentamicin, imipenem and meropenem with DAT or DATS, determined according to the fractional inhibitory concentration index, showed synergic or additive effects. These results suggest that garlic oil, DAT and DATS may have potential for the prevention or treatment of nosocomial, antibiotic-resistant bacterial infections.

Allyl Compounds↗

Antimicrobial susceptibility of Pseudomonas aeruginosa: results of a UK survey and evaluation of the British Society for Antimicrobial Chemotherapy disc susceptibility test.

A survey was conducted in 1999, first to establish the prevalence of antibiotic resistance among clinical isolates of Pseudomonas aeruginosa in the UK and secondly to test whether the use of the standardized British Society for Antimicrobial Chemotherapy (BSAC) disc testing method improved the accuracy of routine susceptibility testing for this organism. Twenty-five hospitals were each asked to collect up to 100 consecutive, clinically significant isolates of P. aeruginosa and to test their susceptibility to amikacin, gentamicin, ceftazidime, imipenem, meropenem, ciprofloxacin, piperacillin and piperacillin/tazobactam using the new BSAC disc method. A total of 2194 isolate reports were available for analysis and 10% of the isolates represented, plus those with unusual resistances, were re-tested centrally for quality control purposes. The zone distributions were essentially unimodal, indicating the absence of major populations with acquired resistance. The results indicated that resistance rates to the beta-lactam, aminoglycoside and quinolone agents tested in P. aeruginosa in the UK remain low (<12%), and were mostly unchanged since a previous survey conducted in 1993. High resistance rates were nevertheless reported for isolates from cystic fibrosis patients. The accuracy of susceptibility testing using the new BSAC disc testing method was better than in previous studies, when Stokes' method was most frequently used. Critically, the proportion of resistant isolates incorrectly reported as susceptible was reduced significantly; nevertheless, depending on the antibiotic, up to 49% of the isolates reported as intermediate or resistant were found susceptible on central re-testing.

Anti-Bacterial Agents↗

Antimicrobial susceptibility patterns of enterococci in intensive care units in Sweden evaluated by different MIC breakpoint systems.

Three hundred and twenty-two (322) clinical isolates were collected from patients admitted to intensive care units (ICUs) at eight Swedish hospitals between December 1996 and December 1998. Of the isolates, 244 (76%) were Enterococcus faecalis, 74 (23%) were Enterococcus faecium and four (1%) were other Enterococcus spp. MICs of ampicillin, imipenem, meropenem, piperacillin/tazobactam, ciprofloxacin, trovafloxacin, clinafloxacin, gentamicin, streptomycin, vancomycin, teicoplanin, quinupristin/dalfopristin, linezolid and evernimicin were determined by Etest. Susceptible and resistant isolates were defined according to the species-related MIC breakpoints of the British Society for Antimicrobial Chemotherapy (BSAC), the National Committee for Clinical Laboratory Standards (NCCLS) and the Swedish Reference Group for Antibiotics (SRGA). Tentative breakpoints were applied for new/experimental antibiotics. Multidrug resistance among enterococci in ICUs is not uncommon in Sweden, particularly among E. faecium, and includes ampicillin resistance and concomitant resistance to fluoroquinolones. Almost 20% of E. faecalis isolates showed high-level resistance to gentamicin and concomitant resistance to fluoroquinolones. Vancomycin-resistant enterococci were only found sporadically. Among the new antimicrobial agents, linezolid and evernimicin showed the best activity against all enterococcal isolates. There was good concordance between the BSAC, NCCLS and SRGA breakpoints in detecting resistance. When applying the SRGA breakpoints for susceptibility, isolates were more frequently interpreted as intermediate. This might indicate earlier detection of emerging resistance using the SRGA breakpoint when the native population is considered susceptible, but with the risk that isolates belonging to the native susceptible population will be incorrectly interpreted as intermediate.

Anti-Bacterial Agents↗

In vitro activity of fosfomycin in combination with various antistaphylococcal substances.

Using the chequerboard technique we studied the in vitro activity of the broad spectrum antibiotic fosfomycin in combination with vancomycin, rifampicin, linezolid, quinupristin/ dalfopristin, cefazolin, meropenem and moxifloxacin against two Staphylococcus epidermidis strains (ATCC 12228, DSM 3269) and five Staphylococcus aureus isolates (ATCC 29213, DSM 683, DSM 46320, GISA 323/93, MRSA 3558/00). The phenomena of 'trailing' and 'skipped wells' did not present a problem. Synergy was the most common effect of all drugs tested in combination with fosfomycin; only combination with vancomycin showed antagonism for two of seven isolates. Using a killing-curve technique fosfomycin showed cidal activity, where increasing the drug concentration above the MIC did not enhance killing velocity. Inhibitory concentrations of vancomycin plus fosfomycin against DSM 46320 caused effects identical to those observed with vancomycin alone. The combination of fosfomycin plus linezolid exerted the bacteriostatic effect found with linezolid alone. Fosfomycin plus quinupristin/dalfopristin exhibited the bactericidal effect found with fosfomycin alone (in contrast to the rapidly bactericidal effect of quinupristin/dalfopristin). Electron microscopy showed that fosfomycin given in combination with linezolid, quinupristin/dalfopristin or moxifloxacin (substances that do not cause morphological alterations when given alone) resulted in 'cauliflower-shaped' distortion as caused by fosfomycin alone. Our in vitro data indicate considerable potential for fosfomycin used in combination with other antistaphylococcal antimicrobials, especially linezolid or quinupristin/dalfopristin.

Anti-Bacterial Agents↗