Effect of basic amino acids on susceptibility to carbapenems in clinical Pseudomonas aeruginosa isolates

We evaluated effects of medium composition, including basic amino acid content and pH, on susceptibility to carbapenems such as imipenem, panipenem and meropenem, in clinical isolates of Pseudomonas aeruginosa. Susceptibility to carbapenems was reduced by basic amino acids in the medium, while susceptibilities to ceftazidime and aztreonam were not. Among carbapenems, susceptibility to panipenem was most sharply reduced by addition of basic amino acids to 1:16 Mueller-Hinton agar (MHA). In 174 of 175 clinical isolates, MICs for carbapenems were affected to different degrees by medium composition. One isolate, in which MICs for carbapenems did not differ between MHA and 1:16 MHA, showed reduced production of porin (OprD). Our results suggest that susceptibility to individual carbapenems, especially panipenem, is difficult to evaluate based on MICs for other carbapenems determined on MHA. For a better prediction of antibiotic efficacy, it may be important to evaluate the susceptibility for each carbapenem individually.     

Antimicrobial susceptibility of bacteria causing urinary tract infections in Latin American hospitals: results from the SENTRY Antimicrobial Surveillance Program (1997)

Objectives: To evaluate the antimicrobial susceptibility patterns among 469 pathogens isolated as a significant cause of urinary tract infections in 10 Latin American medical centers.
Methods: Consecutively collected isolates were susceptibility tested by broth microdilution methods, and selected isolates were characterized by molecular typing methods.
Results: Escherichia coli and Klebsiella spp. isolates revealed high rates of resistance to broad-spectrum penicillins and to fluoroquinolones. Ceftazidime MICs of ≥2 mg/L, suggesting the production of extended-spectrum β-lactamases (ESBLs), were observed in 37.7% of K. pneumoniae and 8.3% of Escherichia coli isolates. Enterobacter spp. isolates were characterized by high resistance rates to ciprofloxacin (35%) and to ceftazidime (45%), but they generally remained susceptible to cefepime (95% susceptible). Pseudomonas aeruginosa and Acinetobacter spp. were highly resistant to ciprofloxacin and ceftazidime. Imipenem was active against 80% of P. aeruginosa and 93% of Acinetobacter spp. isolates.
Conclusions: Our results demonstrate a high level of resistance to various classes of antimicrobial agents among isolates causing nosocomial urinary tract infections in Latin American hospitals. Clonal dissemination of ESBL-producing K. pneumoniae strains was infrequent.     

The Comparative In Vitro Activity of FK-037 (Cefoselis), a New Broad-Spectrum Cephalosporin

FK-037, a new parenteral cephalosporin, was active against clinical isolates of both gram-positive and gram-negative aerobic bacteria. The activity of FK-037 was similar to that of cefpirome and cefepime and generally superior to that of ceftazidime. Methicillin-resistant staphylococci were less susceptible than methicillin-sensitive isolates but most remained within the sensitive range. All streptococci and Enterobacteriaceae were sensitive as were the majority of isolates of Acinetobacter spp. and Pseudomonas aeruginosa. There was resistance amongst other non-fermentative gram-negative bacilli but this was species specific and most pseudomonads were sensitive, with resistance seen in Stenotrophomonas maltophilia, Alcaligenes spp. and Flavobacterium spp.     

In vitro comparative activity of 4 beta-lactams against Pseudomonas aeruginosa. Role of cefoperazone

The authors compared in vitro activity of cefoperazone, piperacillin, carbenicillin and ceftazidime against 100 clinical isolates of Pseudomonas aeruginosa. Minimal inhibitory concentrations (MICs) were determined by the agar dilution method. Minimal bactericidal concentrations were measured using microplates. Eighty-two percent of strains were susceptible or intermediate to cefoperazone, with MICs less than 30 micrograms/ml. The geometric mean of the MICs of cefoperazone was 14.3 micrograms/ml; MIC 50 was 6.9 micrograms/ml. Activity of cefoperazone was comparable to that of carbenicillin, to which eighty-three percent of strains were susceptible, with MICs less than or equal to 128 micrograms/ml. Ceftazidime was also highly active against Pseudomonas, with a mean MIC of 3.29 micrograms/ml. MIC 50 was 1.9 micrograms/ml. Piperacillin was slightly less active, with MICs less than or equal to 32 micrograms/ml for 35% of strains. Cefoperazone exhibited bactericidal activity on 50% of strains at a 28.8 micrograms/ml concentration. These values must be interpreted with caution as they do not allow for the phenomenon of regrowth of Pseudomonas aeruginosa due to the presence of microaggregates of the bacteria in vivo despite bactericidal concentrations of antibiotic.     

Antimicrobial activity of doripenem and other carbapenems against gram-negative pathogens from Korea

A total of 950 gram-negative bacterial isolates from patients with bacteremia and urinary tract infections were collected from tertiary-care hospitals in Korea. In vitro antimicrobial susceptibility testing was performed using broth microdilution test according to Clinical and Laboratory Standards Institute protocol. In general, carbapenems such as doripenem, imipenem, and meropenem were very active against Enterobacteriaceae, Moraxella catarrhalis, Pseudomonas aeruginosa, and Acinetobacter sp. isolates. Doripenem was more potent than imipenem against most Enterobacteriaceae species except Proteus spp. based on minimum inhibitory concentration (MIC)(50) and MIC(90). In addition, doripenem displayed similar activity to meropenem but was superior to imipenem against ceftazidime-resistant Enterobacteriaceae isolates. For P. aeruginosa and Acinetobacter spp. isolates, MIC(50)s of doripenem were 1 and 0.5 mg/L, respectively, which were the same with those of meropenem but two- to fourfold lower than those of imipenem (both 2 mg/L). On the basis of the in vitro data, we conclude that doripenem has equivalent or more activity than other carbapenems such as imipenem and meropenem against most gram-negative pathogens from Korea. Thus, doripenem may be a promising new antimicrobial agent for the treatment of infections caused by gram-negative pathogens in Korea.     

Evaluation of the VITEK 2 system for the identification and susceptibility testing of three species of nonfermenting gram-negative rods frequently isolated from clinical samples

VITEK 2 is a new automatic system for the identification and susceptibility testing of the most clinically important bacteria. In the present study 198 clinical isolates, including Pseudomonas aeruginosa (n = 146), Acinetobacter baumannii (n = 25), and Stenotrophomonas maltophilia (n = 27) were evaluated. Reference susceptibility testing of cefepime, cefotaxime, ceftazidime, ciprofloxacin, gentamicin, imipenem, meropenem, piperacillin, tobramycin, levofloxacin (only for P. aeruginosa), co-trimoxazole (only for S. maltophilia), and ampicillin-sulbactam and tetracycline (only for A. baumannii) was performed by microdilution (NCCLS guidelines). The VITEK 2 system correctly identified 91.6, 100, and 76% of P. aeruginosa, S. maltophilia, and A. baumannii isolates, respectively, within 3 h. The respective percentages of essential agreement (to within 1 twofold dilution) for P. aeruginosa and A. baumannii were 89.0 and 88.0% (cefepime), 91.1 and 100% (cefotaxime), 95.2 and 96.0% (ceftazidime), 98.6 and 100% (ciprofloxacin), 88.4 and 100% (gentamicin), 87.0 and 92.0% (imipenem), 85.0 and 88.0% (meropenem), 84.2 and 96.0% (piperacillin), and 97.3 and 80% (tobramycin). The essential agreement for levofloxacin against P. aeruginosa was 86.3%. The percentages of essential agreement for ampicillin-sulbactam and tetracycline against A. baumannii were 88.0 and 100%, respectively. Very major errors for P. aeruginosa (resistant by the reference method, susceptible with the VITEK 2 system [resistant to susceptible]) were noted for cefepime (0.7%), cefotaxime (0.7%), gentamicin (0.7%), imipenem (1.4%), levofloxacin (2.7%), and piperacillin (2.7%) and, for one strain of A. baumannii, for imipenem. Major errors (susceptible to resistant) were noted only for P. aeruginosa and cefepime (2.0%), ceftazidime (0.7%), and piperacillin (3.4%). Minor errors ranged from 0.0% for piperacillin to 22.6% for cefotaxime against P. aeruginosa and from 0.0% for piperacillin and ciprofloxacin to 20.0% for cefepime against A. baumannii. The VITEK 2 system provided co-trimoxazole MICs only for S. maltophilia; no very major or major errors were obtained for co-trimoxazole against this species. It is concluded that the VITEK 2 system allows the rapid identification of S. maltophilia and most P. aeruginosa and A. baumannii isolates. The VITEK 2 system can perform reliable susceptibility testing of many of the antimicrobial agents used against P. aeruginosa and A. baumannii. It would be desirable if new versions of the VITEK 2 software were able to determine MICs and the corresponding clinical categories of agents active against S. maltophilia.     

Multicentre study of the in vitro activity of cefepime, a broad-spectrum cephalosporin, compared to other broad-spectrum agents

The in vitro activity of cefepime was compared to that of a range of other broad-spectrum agents, using a gradient diffusion MIC method, against 995 recent clinical isolates of Enterobacteriaceae, Pseudomonas aeruginosa, other nonfermentative gram-negative bacilli, staphylococci (except oxacillin-resistant Staphylococcus aureus), streptococci, enterococci, and aerobic gram-positive bacilli. Cefepime had excellent activity against Enterobacteriaceae, including eight presumptive extended-spectrum beta-lactamase producers and 33 stably derepressed mutants of natural cephalosporinases. Activity against Pseudomonas aeruginosa was similar to ceftazidime and superior to cefpirome. Its activity against gram-positive cocci was also good, being more active against staphylococci and only slightly less active against streptococci than ceftriaxone. Cefepime maintained activity against bacteria resistant to aminoglycosides and ciprofloxacin. Enterococci, Bacillus species, Burkholderia cepacia and Stenotrophomonas maltophilia were predicably resistant. Cefepime has a spectrum of activity almost as broad as that of the carbapenems.     

Antimicrobial resistance patterns among aerobic Gram-negative bacilli isolated from patients in intensive care units: results of a multicenter study in Russia

Objective: To determine the antimicrobial resistance patterns among aerobic Gram-negative bacilli isolated from patients in intensive care units (ICUs) in different parts of Russia.
Methods: During 1995–96, 10 Russian hospitals from different geographic areas were asked to submit 100 consecutive Gram-negative isolates from patients with ICU-acquired infections. Minimal inhibitory concentrations (MICs) of 12 antimicrobials were determined by Etest and results were interpreted according to National Committee for Clinical Laboratory Standards (NCCLS) guidelines.
Results: In total, 1005 non-duplicate strains were obtained from 863 patients. The most common species were Pseudomonas aeruginosa (28.8%), Escherichia coli (21.4%), Klebsiella pneumoniae (16.7%), Proteus mirabilis (9.7%), Enterobacter spp. (8.2%) and Acinetobacter spp. (7.7%). High levels of resistance were seen to second- and third-generation cephalosporins, ureidopenicillins, β-lactam/β-lactamase inhibitor combinations and gentamicin. The most active agents were imipenem (no resistance in Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Enterobacter spp. and Acinetobacter spp., 7% resistance in Pseudomonas aeruginosa ), amikacin (7% resistance in Pseudomonas aeruginosa and Acinetobacter spp., 4% in Enterobacter spp., 1% in Escherichia coli and Proteus mirabilis, no resistance in Klebsiella pneumoniae ) and ciprofloxacin (15% resistance in Pseudomonas aeruginosa, 5% in Enterobacter spp. and Proteus mirabilis, 2% in Klebsiella pneumoniae, 1% in Escherichia coli ).
Conclusions: Second- and third-generation cephalosporins, ureidopenicillins, β-lactam/β-lactamase inhibitor combinations and gentamicin cannot be considered as reliable drugs for empirical monotherapy for aerobic Gram-negative infections in ICUs in Russia.     

Antibacterial activity of carumonam and cefpirome on hospital strains resistant to gentamicin and cephalothin: comparison with other beta-lactam antibiotics, new fluoroquinolones, aminoglycosides and other antibiotics

The antibacterial in vitro activity of carumonam, a new monobactam, and cefpirome, a new cephalosporin, was studied on 483 hospital strains resistant to gentamicin and cephalothin, in comparison with amikacin, azlocillin, aztreonam, cefmenoxim, cefoperazone, cefotaxim, cefsulodin (for Pseudomonas), ceftazidime, ceftriaxone, cefuroxim, chloramphenicol, ciprofloxacin, doxycycline, enoxacin, netilmicin, norfloxacin, pefloxacin, piperacillin, rifampicin, tobramycin and trimethoprim. In general the two compounds have a very good in vito activity on Enterobacteriaceae but are less active on non-fermenting microorganisms. For the Enterobacteriaceae the minimal inhibitory concentrations 90% for carumonam was less than or equal to 1.1 mg/l excepted for Enterobacter spp. (43,6 mg/l) and M. morganii (56.8 mg/l) . All the Enterobacteriaceae are susceptible to cefpirome (minimal inhibitory concentrations 90% less than or equal to 5.3 mg/l). The activity of carumonam and cefpirome on Enterobacteriaceae is comparable with that of the third generation cephalosporins. Carumonam is more active than cefpirome and other beta-lactams, ceftazidime excepted, on Pseudomonas aeruginosa and Pseudomonas spp. On the other hand, both compounds reveal to have only a low activity on the other non-fermenters which minimal inhibitory concentrations 90% values of 115.4 mg/l for carumonam and 32.0 mg/l for cefpirome.     

4-h Identification of Pseudomonas aeruginosa with 9-chloro-9-(4-diethylaminophenyl)-10-phenylacridan.

A total of 361 gram-negative bacilli were evaluated for their ability to grow in the presence of 9-chloro-9-(4-diethylaminophenyl)-10-phenylacridan. The minimum time required for the production of visual turbidity in brain heart infusion broth was determined to be 4 h in shake cultures at 35 degrees C. The minimal inhibitory concentration (MIC) of 9-chloro-9-(4-diethylaminophenyl)-10-phenylacridan in brain heart infusion broth was determined for 174 isolates. The MICs for all 40 Pseudomonas aeruginosa isolates tested were greater than 50 micrograms/ml. The MICs for the other 53 pseudomonads tested were less than or equal to 5 micrograms/ml. Among 81 other gram-negative bacilli tested, the MICs for 4 were 15 micrograms/ml, the MICs for 15 were 10 micrograms/ml, the MICs for 21 were 5 micrograms/ml, and the MICs for 41 were less than or equal to 1 micrograms/ml. Based on these data, 361 gram-negative bacilli were inoculated into brain heart infusion broth containing 15 micrograms of 9-chloro-9-(4-diethylaminophenyl)-10-phenylacridan per ml and incubated on a shaker for 4 h. The only bacteria that produced visual turbidity were identified as P. aeruginosa (170 of 170 isolates).     

Development of resistance in Pseudomonas aeruginosa obtained from patients with cystic fibrosis at different times

Objective  Determination of the extent of changes in quantitative resistance in Pseudomonas aeruginosa isolates from patients with cystic fibrosis over a period of approximately 2 years.
Methods  Three hundred and ninety nine isolates of P. aeruginosa collected from 34 pediatric patients in the period between April 1994 and April 1996 were investigated. During the 2 years the children were treated with a combination of a betalactam and an aminoglycoside, approximately every 3 months. In between they received ciprofloxacin orally, when required. The minimal inhibitory concentrations (MICs) of 38 clones of P. aeruginosa defined by different patterns in macrorestriction analysis (pulse field gel electrophoresis, PFGE) were established for 12 antibiotics: gentamicin, amikacin, tobramycin, ciprofloxacin, levofloxacin, moxifloxacin, trovafloxacin, imipenem, meropenem, ceftazidime, cefepime, and piperacillin by means of broth microdilution tests according to DIN 58940.
Results  Twenty-four of the 38 clones developed increased MIC values during the time of observation especially for aminoglycosides and quinolones. Comparatively less affected were ceftazidime, imipenem and meropenem. An association between the number of the intravenous treatment courses and the increase of the MIC values could not be verified.
Conclusions  A trend towards an increase of the MICs against antipseudomonal agents was observed over a limited period of time. It is necessary to prevent this development possibly by employing suitable combinations of antibiotics and the introduction of new substances.     

Cation concentration variability of four distinct Mueller-Hinton agar brands influences polymyxin B susceptibility results

Polymyxins have been the only alternative therapeutic option for the treatment of serious infections caused by multidrug-resistant Acinetobacter baumannii or Pseudomonas aeruginosa isolates. For this reason, it is of crucial importance that susceptibility tests provide accurate results when testing these drug-pathogen combinations. In this study, the effect of cation concentration variability found on different commercial brands of Mueller-Hinton agar (MHA) for testing polymyxin B susceptibility was evaluated. The polymyxin B susceptibilities determined using Etest and disk diffusion were compared to those determined by the CLSI reference broth microdilution method. In general, the polymyxin B MIC values were higher when determined by Etest than when determined by broth microdilution against both A. baumannii and P. aeruginosa isolates. A high very major error rate (10%) was observed, as well as a trend toward lower MICs, compared to those determined by broth microdilution when the Merck MHA was tested by Etest. Poor essential agreement rates (10 to 70%) were observed for P. aeruginosa when all MHA brands were tested by Etest. Although an excellent categorical agreement rate (100%) was seen between the disk diffusion and broth microdilution methods for P. aeruginosa, larger zones of inhibition were shown obtained using the Merck MHA. The high cation concentration variability found for the MHA brands tested correlated to the low accuracy, and discrepancies in the polymyxin B MICs were determined by Etest method, particularly for P. aeruginosa isolates.     

Molecular surveillance of European quinolone-resistant clinical isolates of Pseudomonas aeruginosa and Acinetobacter spp. using automated ribotyping

Nosocomial isolates of Pseudomonas aeruginosa and Acinetobacter spp. exhibit high rates of resistance to antibiotics and are often multidrug resistant. In a previous study (D. Milatovic, A. Fluit, S. Brisse, J. Verhoef, and F. J. Schmitz, Antimicrob. Agents Chemother. 44:1102-1107, 2000), isolates of these species that were resistant to sitafloxacin, a new advanced-generation fluoroquinolone with a high potency and a broad spectrum of antimicrobial activity, were found in high proportion in 23 European hospitals. Here, we investigate the clonal diversity of the 155 P. aeruginosa and 145 Acinetobacter spp. sitafloxacin-resistant isolates from that study by automated ribotyping. Numerous ribogroups (sets of isolates with indistinguishable ribotypes) were found among isolates of P. aeruginosa (n = 34) and Acinetobacter spp. (n = 16), but the majority of the isolates belonged to a limited number of major ribogroups. Sitafloxacin-resistant isolates (MICs > 2 mg/liter, used as a provisional breakpoint) showed increased concomitant resistance to piperacillin, piperacillin-tazobactam, ceftriaxone, ceftazidime, amikacin, gentamicin, and imipenem. The major ribogroups were repeatedly found in isolates from several European hospitals; these isolates showed higher levels of resistance to gentamicin and imipenem, and some of them appeared to correspond to previously described multidrug-resistant international clones of P. aeruginosa (serotype O:12) and Acinetobacter baumannii (clones I and II). Automated ribotyping, when used in combination with more discriminatory typing methods, may be a convenient library typing system for monitoring future epidemiological dynamics of geographically widespread multidrug-resistant bacterial clones.     

Antimicrobial resistance among Gram-negative bacilli causing infections in intensive care unit patients in the United States between 1993 and 2004

During the 12-year period from 1993 to 2004, antimicrobial susceptibility profiles of 74,394 gram-negative bacillus isolates recovered from intensive care unit (ICU) patients in United States hospitals were determined by participating hospitals and collected in a central location. MICs for 12 different agents were determined using a standardized broth microdilution method. The 11 organisms most frequently isolated were Pseudomonas aeruginosa (22.2%), Escherichia coli (18.8%), Klebsiella pneumoniae (14.2%), Enterobacter cloacae (9.1%), Acinetobacter spp. (6.2%), Serratia marcescens (5.5%), Enterobacter aerogenes (4.4%), Stenotrophomonas maltophilia (4.3%), Proteus mirabilis (4.0%), Klebsiella oxytoca (2.7%), and Citrobacter freundii (2.0%). Specimen sources included the lower respiratory tract (52.1%), urine (17.3%), and blood (14.2%). Rates of resistance to many of the antibiotics tested remained stable during the 12-year study period. Carbapenems were the most active drugs tested against most of the bacterial species. E. coli and P. mirabilis remained susceptible to most of the drugs tested. Mean rates of resistance to 9 of the 12 drugs tested increased with Acinetobacter spp. Rates of resistance to ciprofloxacin increased over the study period for most species. Ceftazidime was the only agent to which a number of species (Acinetobacter spp., C. freundii, E. aerogenes, K. pneumoniae, P. aeruginosa, and S. marcescens) became more susceptible. The prevalence of multidrug resistance, defined as resistance to at least one extended-spectrum cephalosporin, one aminoglycoside, and ciprofloxacin, increased substantially among ICU isolates of Acinetobacter spp., P. aeruginosa, K. pneumoniae, and E. cloacae.     

PCR detection of metallo-beta-lactamase gene (blaIMP) in gram-negative rods resistant to broad-spectrum beta-lactams.

We applied PCR to the rapid detection of the metallo-beta-lactamase gene, blaIMP, in clinically isolated gram-negative rods. A total of 54 high-level ceftazidime-resistant strains (MICs, > 128 micrograms/ml) were subjected to PCR analyses with the blaIMP-specific primers, since the blaIMP-bearing clinical isolates tested in our previous study always demonstrated high-level resistance to ceftazidime. Twenty-two blaIMP-positive strains including 9 Pseudomonas aeruginosa, 9 Serratia marcescens, 2 Alcaligenes xylosoxidans, 1 Pseudomonas putida, and 1 Klebsiella pneumoniae strains were newly identified from 18 different hospitals in Japan. These strains were mostly isolated from urine samples and showed high-level resistance to almost every cephem, while their levels of resistance to carbapenems were diverse. The PCR analyses with novel integrase gene-specific (intI3) and acc(6')-Ib gene-specific primers suggested that the integron structure found in a large plasmid harbored by S. marcescens AK9373 was also well conserved among blaIMP-positive strains. These results imply that the blaIMP gene cassettes have been dispersing into various gram-negative rods with the help of the newly identified integron element. Thus, the PCR-aided rapid detection will be helpful for the early recognition of emerging blaIMP-positive clinical isolates which demonstrate consistent resistance to beta-lactams.     

Imipenem susceptibility testing with a commercially prepared dry-format microdilution tray.

MICs of imipenem, concurrently generated in commercially prepared microdilution trays containing predried antibiotic dilutions (Sensititre), and in a standard agar dilution assay (as recommended by the National Committee for Clinical Laboratory Standards, Villanova, Pa.), were within +/- 1 twofold dilution for 94% of 226 bacterial isolates. Imipenem biological activity remained stable over 5 months of tray storage at room temperature against Pseudomonas aeruginosa ATCC 27853. Activity of imipenem was shown by microdilution testing with 890 clinical isolates to be high, with only 4% of isolates having MICs of greater than or equal to 16 micrograms/ml (in vitro resistance).     

P594 Intracellular killing of drug-resistant Staphylococcus aureus by different antibiotics

Objectives:  The aim of this study was to assess the carbapenem susceptibility of four nosocomial pathogens and to evaluate the reliability of the susceptibility results determined by E-test and disc diffusion (DD) methods.
Methods:  Escherichia coli ( n  = 73), Klebsiella pneumoniae ( n  = 60), Pseudomonas aeruginosa ( n  = 70) and Acinetobacter spp. ( n  = 70) isolated from nosocomial infections in 2002–2003 were included in the study. Thirty-five per cent of the strains were isolated from intensive care units. After determining antimicrobial susceptibility against imipenem and meropenem by DD (10  μ g; Oxoid, UK) and Etest (AB Biodisk, Solna, Sweden) methods, the results were categorised as susceptible (S), intermediate (I) and resistant (R) according to the NCCLS criteria. For statistical analyses, the intermediate group was included in the resistant category because of the low numbers of bacteria in the former group.
Results:  None of E. coli or K. pneumoniae strains were resistant to carbapenems, whereas, resistance reached up to 59.0% in Acinetobacter spp. and P. aeruginosa isolates. By either method, the pattern of the susceptibility of the four bacteria was not statistically significantly different for imipenem vs. meropenem. Total agreement of DD and E-test methods for susceptibility to imipenem was 95.7%, and 90.0% in Acinetobacter spp. and P. aeruginosa , respectively; and susceptibility to meropenem was 90.0% for both bacteria. However, the difference of the results obtained by either method was statistically significant for Acinetobacter spp.
Conclusion:  Study results suggest a high resistance rate for Acinetobacter spp. and P. aeruginosa strains against carbapenem antibiotics in our hospital. Further studies are needed to clarify whether E-test should be used to confirm meropenem resistance of Acinetobacter spp. determined by DD method.     

Biochemical characterization of a novel extended-spectrum beta-lactamase from Pseudomonas aeruginosa 802

Pseudomonas aeruginosa 802 was isolated at Rabta hospital in Tunis and was resistant to extended-spectrum cephalosporins and aztreonam. It produced a pI 7.6 extended-spectrum beta-lactamase (ESBL). The ESBL, named LBT 802, was purified to homogeneity by filtration on Sephadex G-75 followed by CM-Sepharose chromatography and high-performance liquid chromatography (HPLC) on a TSK-gel SP-5PW column. The LBT 802 enzyme had a molecular mass of 30 kDa. It showed a broad-substrate profile by hydrolyzing benzylpenicillin, ampicillin, cephalothin, cephaloridine, cefotaxime, ceftriaxone, and cefpirome but not ceftazidime, cefoxitin, imipenem, or aztreonam. The highest hydrolytic efficiency (Vmax/Km) was obtained for ampicillin, cephalothin, cephaloridine, and benzylpenicillin. Among extended-spectrum cephalosporins the best substrate was ceftriaxone followed by cefotaxime and cefpirome. LBT 802 activity was inhibited by clavulanic acid, sulbactam, imipenem, cefoxitin, and aztreonam. It showed its lowest Ki values for clavulanic acid, imipenem and sulbactam.     

In vitro antimicrobial activity of diethyldithiocarbamate and dimethyldithiocarbamate against methicillin-resistant Staphylococcus

Staphylococcus aureus has appeared which is highly resistant to both methicillin and aminoglycosides. Current therapy involves long-term intravenous therapy of vancomycin. Since vancomycin is currently the only drug used to treat these patients, there is a need to develop additional antimicrobial therapy. The in vitro antimicrobial effect of the metal chelator, diethyldithiocarbamate (DDTC) and its structural analog dimethyldithiocarbamate (DMTC) were investigated. Both DDTC and DMTC were effective against S. aureus including methicillin-resistant S. aureus (MRSA). By agar diffusion, DDTC at 10 micrograms per disk produced zone sizes of 12 to 21 mm and at 100 micrograms per disk produced zone sizes of 26 to 34 mm against MRSA. The DMTC produced slightly greater zone sizes against MRSA of 16 to 24 mm and 24 to 37 mm for 10 micrograms per disk and 100 micrograms per disk, respectively. The minimum inhibitory concentration (MIC) for DMTC against MRSA was 6 micrograms per ml. Both DDTC and DMTC were also effective against enterococci, Proteus mirabilis, Klebsiella pneumoniae, Klebsiella oxytoca, Escherichia coli, Enterobacter cloacae, Enterobacter aerogenes, Salmonella species, Serratia marcescens and Citrobacter freundii at 100 micrograms per disk. The MICs of DMTC for Klebsiella pneumoniae, Klebsiella oxytoca, Escherichia coli, Salmonella and Citrobacter freundii were approximately 128 micrograms per ml while the MICs for Proteus vulgaris, Proteus mirabilis, Pseudomonas aeruginosa and Serratia marcescens was greater than or equal to 256 micrograms per ml. In addition, DMTC was synergistic with gentamicin against MRSA and coagulase-negative staphylococcus species, Enterobacter cloacae, Klebsiella pneumoniae and Pseudomonas aeruginosa. Additive and synergistic effects of DMTC were displayed with gentamicin against S. aureus including methicillin-resistant S. aureus.(ABSTRACT TRUNCATED AT 250 WORDS)     

Antimicrobial activity of the new carbapenem biapenem compared to imipenem,meropenem and other broad-spectrum beta-lactam drugs

The in vitro activity of biapenem was compared to that of imipenem, meropenem and other broad-spectrum beta-lactams. A total of 716 isolates from recent cases of clinical septicemia and an additional 137 stock strains possessing known beta-lactamases or other well-characterized resistance mechanisms were tested. The minimal concentrations inhibiting 90 % of strains (MIC90) ofEnterobacteriaceae species were for biapenem 0.03 to 1 mg/l and for imipenem 0.25 to 2 mg/l. No member of theEnterobacteriaceae was found to be resistant to biapenem. Biapenem and meropenem were the most active drugs againstPseudomonas aeruginosa, with an MIC90 of 1 mg/l. Biapenem was more active than ceftazidime against most gram-negative and gram-positive bacteria tested. Biapenem was as potent as imipenem against anaerobic bacteria (includingBacteroides fragilis), with an MIC90 of 0.25 mg/l. High MICs of biapenem were demonstrated forXanthomonas maltophilia, oxacillin-resistantStaphylococcus spp. andEnterococcus spp. These species have demonstrated resistance to other carbapenems and to most of the newer cephalosporins. The results of this study, coupled with previously documented favorable qualities of biapenem, endorse further investigation of this broad-spectrum antibacterial agent for clinical use.