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Antimicrobial susceptibilities among clinical isolates of extended-spectrum cephalosporin-resistant Gram-negative bacteria in a Taiwanese University Hospital.

Infections caused by Gram-negative bacteria with resistance to extended-spectrum cephalosporins require the identification of effective alternative antimicrobial therapy. To determine the role of other pre-existing or currently available antimicrobial agents in treating infections caused by these multidrug-resistant pathogens, we evaluated the in vitro susceptibilities of these agents in 411 non-duplicate isolates of extended-spectrum cephalosporin-resistant Gram-negative bacteria recovered between January 1999 and December 1999 in a major teaching hospital in Taipei, Taiwan. These isolates included cefotaxime-resistant (MICs > or = 2 mg/L) Escherichia coli (66 isolates) and Klebsiella pneumoniae (77 isolates); cefotaxime-resistant (MICs > or = 64 mg/L) Enterobacter cloacae (59 isolates), Serratia marcescens (52 isolates) and Citrobacter freundii (52 isolates); and ceftazidime-resistant (MICs > or = 64 mg/L) Pseudomonas aeruginosa (50 isolates) and Acinetobacter baumannii (55 isolates). Overall, carbapenems (imipenem and meropenem) had good activity against the cefotaxime-resistant Enterobacteriaceae tested (>90% of isolates were susceptible). However, carbapenems had limited activity against the ceftazidime-resistant P. aeruginosa (only 4% of isolates were susceptible) and A. baumannii (51-56% of isolates were susceptible). Among the E. coli and K. pneumoniae isolates tested, 33.3% and 58.4%, respectively, exhibited extended-spectrum beta-lactamase phenotype, determined by the double disc method. Over 80% of cefotaxime-resistant E. cloacae and C. freundii were susceptible to cefepime, but this agent had limited activity against other bacteria tested. Susceptibilities of these isolates to ciprofloxacin varied, ranging from 25% for A. baumannii to 92% for E. cloacae. Newer fluoroquinolones (moxifloxacin and trovafloxacin) had equal or less activity against these organisms, except for A. baumannii for which their MIC(90)s (8-16 mg/L) were four- to 16-fold less than that of ciprofloxacin (MIC(90) 128 mg/L).

Cefotaxime↗

Antibiotic resistance among clinical isolates of Acinetobacter in the UK, and in vitro evaluation of tigecycline (GAR-936).

A survey was conducted of the antimicrobial susceptibilities of 595 Acinetobacter spp. isolated from routine clinical specimens in 54 sentinel laboratories throughout the UK during 2000. Isolates of the Acinetobacter baumannii complex (genomic groups 2, 3 and 13TU; n = 443) were distinguished from other genomic groups (n = 152) by PCR fingerprinting of tDNA spacer regions. MICs of amikacin, cefotaxime, ceftazidime, ciprofloxacin, colistin, gentamicin, imipenem, meropenem, minocycline, piperacillin, piperacillin/tazobactam, rifampicin, sulbactam and tetracycline were determined on IsoSensitest agar and interpreted, wherever possible, using BSAC breakpoints. Tigecycline (GAR-936), a new glycylcycline, was also tested. Resistance to cephalosporins, aminoglycosides and ciprofloxacin was widespread, but carbapenems, colistin, sulbactam, minocycline and tigecycline were each active against >80% of the isolates. Isolates of A. baumannii were more often resistant to cefotaxime, ceftazidime, piperacillin, piperacillin/tazobactam, ciprofloxacin, gentamicin and tetracyclines than those belonging to other genomic groups, but were less often resistant to colistin; no significant differences between genomic groups were noted in the susceptibilities to amikacin, carbapenems, rifampicin or sulbactam. The relative activities of the tetracyclines were minocycline > tigecycline > tetracycline. Thirteen carbapenem-resistant isolates (MICs > or =8 mg/L; 2.2%) were received from six centres; four centres sent single isolates; one sent three and one sent six. An allele of bla(IMP) was detected in one of these isolates, but the other 12 isolates either had carbapenemase-independent resistance, or undetectable carbapenemase activity combined with other resistance mechanisms. In conclusion, carbapenems, colistin and minocycline retained greatest activity against the Acinetobacter isolates collected. Tigecycline was less active than minocycline, but both agents overcame most tetracycline resistance.

Acinetobacter↗

Evaluation of MIDITECH automated colorimetric MIC reading for antimicrobial susceptibility testing.

The MIDITECH colorimetric susceptibility test with automated reading is a modification of the standard broth microdilution method that uses a 3-(4,5-dimetylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) dye for detecting viable bacteria. The method can be applied to non-fastidious aerobic Gram-negative bacteria, staphylococci and Enterococcus faecalis. To assess the reliability of this method, we compared susceptibility data obtained by this test with standard NCCLS microdilution assay results. For this purpose, 15 antibiotics and a well characterized set of 527 Gram-negative and Gram-positive bacterial isolates collected and stored at the Division of Infectious Diseases and Chemotherapy (Vienna General Hospital, Austria), yielding 5751 organism-antibiotic combinations, were analysed in duplicate. The overall essential agreement (+/-1 log(2) dilution) between the MIDITECH and NCCLS methods was 96.18 +/- 0.67%. The colorimetric assay compared with the reference method produced MICs < or = 2 log(2) dilutions and > or = 2 log(2) dilutions in 2.34% and 1.48% comparisons, respectively. For 326 Gram-negative bacteria, the absolute interpretative agreement of both methods ranged from 87.12% for ampicillin-sulbactam to 99.85% for meropenem (mean 94.86%); 417 (4.92%) minor, three (0.05%) major and 15 (0.63%) very major errors were found. For 127 staphylococci and 74 E. faecalis isolates, the absolute interpretative agreement ranged from 90.80% for ciprofloxacin to 100% for vancomycin and linezolid (mean 96.96%); 81 (2.77%) minor, three (0.15%) major and eight (0.83%) very major errors were found. For most of the clinically important aerobically growing pathogens, the MIDITECH colorimetric test provided reliable quantitative susceptibility data. The main advantage of this method is simple performance, automated reading and data processing without expensive investments.

Colorimetry↗

Characterization of blaOXA-542-mediated carbapenem resistance in Acinetobacter baumannii.

BACKGROUND: Carbapenem-resistant Acinetobacter baumannii (CRAB) causes multiple anatomical site infections, representing a significant public health threat. AIM: This study reports the isolation and characterization of a carbapenem-resistant A. baumannii harbouring blaOXA-542, followed by a comprehensive investigation of its antimicrobial resistance mechanisms and genomic characteristics. METHODS: Firstly, antimicrobial susceptibility testing was performed using the broth microdilution method. Subsequently, whole-genome sequencing was employed to identify and characterize the resistance and virulence determinants. The functional validation of resistance mechanisms was performed by gene knockdown and construction of expression vectors. The fitness cost of &#x3b2;-lactamase expression was identified by a bacterial growth kinetic test. Molecular docking was utilized to predict potential binding sites of &#x3b2;-lactamase and carbapenems. Finally, the genetic characteristics of the isolates were analysed through comparative genomics analyses and phylogenetic tree construction. RESULTS AND CONCLUSIONS: The results demonstrated that blaOXA-542 confers resistance to carbapenem and penicillin in A. baumannii and Escherichia coli while exhibiting no significant impact on cephalosporins. The ability of blaOXA-542 to hydrolyze meropenem was further confirmed by modified carbapenem inactivation assay (mCIM). Expression of blaOXA-542 in E. coli BL21 showed no significant growth rate alteration. Comparative analysis of the blaOXA-542 genetic environment revealed a close association with Acinetobacter pitti. This study reports the emergence of blaOXA-542-mediated carbapenem and penicillin resistance in a novel A. baumannii lineage (ST2795Pas/ST3464Oxf), highlighting the urgent need for rational antibiotic use against specific pathogens.

Acinetobacter baumannii↗

Phenotype-genotype discordance in antimicrobial resistance profiles of Gram-negative uropathogens recovered from catheter-associated urinary tract infections in Egypt.

OBJECTIVES: Catheter-associated urinary tract infections (CAUTIs) are among the most common healthcare-associated infections in low- and middle-income countries (LMICs), but there are few resistome data available for relevant uropathogens. The goal of this study was to characterize the antimicrobial resistance (AMR) phenotypes and genotypes of a large collection of Gram-negative bacteria recovered from CAUTIs in a hospital in Mansoura, Egypt. METHODS: Phenotypic AMR profiles and whole-genome sequence data were generated for 132 isolates. Resistomes were predicted using ResFinder, CARD and AMRFinder. Similarity of uropathogen genomic data was determined using sourmash (kmer signatures). Escherichia coli genomic data were subject to a pangenome analysis using Panaroo. RESULTS: Sixty-seven E. coli (Phylogroup B2; 53.7%, 36/67), 14 Pseudomonas aeruginosa, 11 Klebsiella pneumoniae, 9 Proteus mirabilis, 8 Providencia spp., 5 Enterobacter hormaechei and 18 rare CAUTI-associated isolates were identified. Several (22/132) isolates were multidrug-resistant, while almost half (62/132) were extensively drug-resistant. Phenotype-genotype discordance was found to be an important consideration in resistome studies in Egypt, with a total concordance of 91% (1115/1225), 85.7% (1273/1485) and 80.5% (1196/1485) for ResFinder, CARD and AMRFinder, respectively. Pseudomonas, at the species level, exhibited the greatest discordance. At the antimicrobial level, meropenem was subject to greatest discordance. New AMR variants were found for Egypt for Pseudomonas (blaOXA-486, blaOXA-488, blaOXA-905, blaIMP-43, blaPDC-35, blaPDC-45, blaPDC-201) and E. coli (blaTEM-176, blaTEM-190). CONCLUSIONS: This study shows that there is phenotype-genotype discordance in AMR profiling among CAUTI isolates, highlighting the need for comprehensive approaches in resistome studies. We also show the genomic diversity of Gram-negative uropathogens contributing to disease burden in a little-studied LMIC setting.

Egypt↗

Anti-anaerobic activity of AZD2563, a new oxazolidinone, compared with eight other agents.

The anti-anaerobic activity of AZD2563, a new oxazolidinone, was compared with that of eight other agents against 201 Gram-positive and 99 Gram-negative anaerobes. MIC(50) and MIC(90) values (mg/L) for the 201 Gram-positive organisms were as follows: AZD2563, 1.0/4.0; linezolid, 1.0/4.0; quinupristin/dalfopristin, 0.5/1.0; amoxicillin, 0.25/1.0; clindamycin, 0.25/8.0; metronidazole, 0.5/>16.0; vancomycin, 0.5/2.0; teicoplanin, 0.125/0.25; and meropenem, 0.06/1.0. AZD2563, linezolid, vancomycin, teicoplanin and quinupristin/dalfopristin were mainly active against Gram-positive anaerobes, with no useful activity against Gram-negative anaerobes.

Anti-Bacterial Agents↗

Metallo-beta-lactamase-producing Enterobacteriaceae isolates in a university hospital in Taiwan: prevalence of IMP-8 in Enterobacter cloacae and first identification of VIM-2 in Citrobacter freundii.

A total of 9082 clinical isolates of Enterobacteriaceae other than Klebsiella spp. collected in 1999 and 2000 at a university hospital in Taiwan were investigated for the production of metallo- beta-lactamases (MBLs). Thirty-six (2.9%) of the 1261 Enterobacter cloacae isolates and one (0.3%) of the 340 Citrobacter freundii isolates were found to carry bla(IMP-8) and bla(VIM-2), respectively, by colony hybridization, PCR and sequence analysis. The IMP-8 producers were recovered from 20 patients and four of them had recently transferred from other hospitals, implying spread of IMP-8-producing E. cloacae among different healthcare settings. Of the 20 non-repetitive IMP-8 producers, 17 (85%) isolates also harboured bla(SHV-12), which was on the same transferable plasmids with bla(IMP-8). The bla(VIM-2)-positive isolate and all non-repetitive bla(IMP-8)-positive isolates appeared susceptible to imipenem (MICs < 8 mg/L) and meropenem (MICs < 4 mg/L), indicating the difficulty in detection of MBLs in Enterobacteriaceae by routine susceptibility testing. Ribotyping of the IMP-8-producing E. cloacae isolates indicated that the dissemination of bla(IMP-8) was due largely to the spread of an epidemic clone, but horizontal transfer among unrelated strains also occurred.

Citrobacter freundii↗

Increase in resistance to new fluoroquinolones from 1998 to 2001 in the Bacteroides fragilis group.

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, especially to certain antibiotics such as clindamycin and cefoxitin. The antimicrobial susceptibilities of 200 strains of the B. fragilis group isolated in 1998 (100) and 2001 (100) from faecal samples of healthy people to anti-anaerobic agents were determined. Meropenem, metronidazole and chloramphenicol showed excellent activity against all isolates. The efficacy of cefoxitin was low, with only 43% of strains isolated in 2001 showing susceptibility. A high prevalence of resistance to clindamycin, reaching 56% of recent isolates, was observed. A significant increase in resistance to new fluoroquinolones, from 0% in 1998 to 12% in 2001, was detected.

Anti-Infective Agents↗

Evaluation of selective and enrichment media for isolation of glycopeptide-resistant enterococci from faecal specimens.

OBJECTIVES: Vancomycin-resistant enterococcus enrichment broth (VEB) and vancomycin-resistant enterococcus selective agar with vancomycin 6 mg/L (VSA) are novel azide-aesculin agar-based media that contain meropenem as an additional selective agent. The media were compared with enterococcosel broth (EB) and enterococcosel agar with vancomycin 6 mg/L (EA) for the isolation of glycopeptide-resistant enterococci (GRE) from routine faecal screening specimens. METHODS: Two hundred and eighteen routine faecal screening specimens from patients at Addenbrooke's Hospital were examined. The majority were from patients on haematology wards (155) or the intensive therapy unit (ITU) (21). Specimens were inoculated on to VSA and EA directly, and after enrichment in VEB and EB, respectively. RESULTS: One hundred and twenty-eight GRE isolates were recovered from 93 (43%) specimens with enterococci carrying vanA or vanB genes. There were no statistically significant differences between media (specimens positive; numbers of GRE isolates) on direct plating on VSA (87; 104) or EA (86; 97) or following 24 h enrichment in VEB (89; 103) or EB (86; 98). There was no significant advantage to enrichment compared with direct plating. Incubation of enrichment broth cultures for only 6 h appeared detrimental. Enterococci with vanC were isolated significantly less frequently from VEB and VSA than from EB and EA. Growth of organisms other than GRE was more common on VSA than on EA. CONCLUSIONS: VEB and VSA were at least as effective as EB and EA for the recovery of GRE from faecal screening specimens, but substantially more non-GRE grew on VSA than on EA. Enrichment culture offered no significant advantages over direct plating.

Agar↗

Multicentre surveillance of Pseudomonas aeruginosa susceptibility patterns in nosocomial infections.

OBJECTIVES: To determine susceptibility rates and patterns in Pseudomonas aeruginosa strains isolated from nosocomial infections. METHODS: Seven hundred and sixteen P. aeruginosa isolates from 40 different hospitals in Belgium and the Grand Duchy of Luxembourg were collected in 1999. RESULTS: Resistance rates varied significantly between hospitals. Of the fluoroquinolones, ciprofloxacin showed least resistance (24%), levofloxacin showed 27.5% resistance and ofloxacin 37.5%. Of the aminoglycosides, amikacin was the most potent antibiotic (10.5% resistance), followed by isepamicin (12%), tobramycin (19.5%) and gentamicin (23.5%). Of the beta-lactam antibiotics, meropenem was the most active (9.5% resistance); piperacillin and piperacillin/tazobactam had, respectively, 24% and 17.5% resistance, ceftazidime 28.5%, cefepime 29.5%, ticarcillin/clavulanic acid 37% and aztreonam 55.5%. MIC distribution curves show the presence of significant subpopulations, with MICs just below breakpoint for many antibiotics. CONCLUSION: Resistance of P. aeruginosa to penicillins, cephalosporins, fluoroquinolones and aminoglycosides varies between hospitals, but is increasing.

Anti-Bacterial Agents↗

Hospital outbreak of multiple clones of Pseudomonas aeruginosa carrying the unrelated metallo-beta-lactamase gene variants blaVIM-2 and blaVIM-4.

OBJECTIVES: The possible contribution of metallo-beta-lactamases in the frequent detection of carbapenem-resistant Pseudomonas aeruginosa isolates in a tertiary Greek hospital in Central Greece was investigated. MATERIALS AND METHODS: All carbapenem-resistant (imipenem- and/or meropenem-resistant) P. aeruginosa isolates recovered from separate patients during a 1 year period in the Clinical Microbiology Laboratory at the University Hospital of Thessaly, Larissa, Greece, were studied for metallo-beta-lactamases. They were tested by Etest MBL, PCR analysis and nucleotide sequencing. DNA fingerprints were obtained by pulsed-field gel electrophoresis (PFGE) of XbaI-digested chromosomal DNA. RESULTS: A blaVIM gene was detected in 47 of the 53 (88.7%) carbapenem-resistant P. aeruginosa isolates. PFGE grouped the blaVIM-positive isolates in six unrelated genotypes; one type included two subtypes. Nucleotide sequencing of the PCR amplicons of a randomly selected isolate from each one of the seven subtypes, detected the variant sequences blaVIM-2 in four and blaVIM-4 in three cases, respectively. They were carried as single gene cassettes or along with an aminoglycoside resistance gene (aacA29a) in class 1 integrons. CONCLUSIONS: These findings suggest that different strains of P. aeruginosa carrying unrelated metallo-beta-lactamase gene variants predominate in our hospital environment.

Anti-Bacterial Agents↗

Properties and potential of ertapenem.

Ertapenem is a carbapenem that shares the activity of imipenem and meropenem against most species, but is less active against non-fermenters. Activity is retained against most strains with AmpC and extended-spectrum beta-lactamases, although resistance can arise if these enzymes are combined with extreme impermeability. Resistance can also be caused by IMP, VIM, KPC and NMC carbapenemases, but again, co-requires impermeability. Although the spread of carbapenemases in the future is a concern, they are currently very rare. Given as a 1 g intravenous (iv) infusion once daily, ertapenem has a plasma half-life of approximately 4 h in healthy volunteers, and a Cmax of 155 mg/L and 13 mg/L for total and free drug, respectively. Excretion is largely renal, divided equally between native drug and an open-ring derivative. Trials show equivalence to piperacillin/tazobactam or ceftriaxone in (a) intra-abdominal infections, (b) community-acquired pneumonia, (c) acute pelvic infections, (d) skin and skin structure infections and (e) complicated urinary tract infections. The USA licence grants all these five indications; the EU licence grants the first three. Further potential uses include home iv therapy, directed therapy against Enterobacteriaceae with AmpC or extended-spectrum cephalosporinases, and tentatively, surgical prophylaxis. Widening the usage of carbapenems raises public health concerns, somewhat allayed by the continued rarity of carbapenemases after 17 years of imipenem use, and by the fact that carbapenemases occur mostly in non-fermenters outside the spectrum of ertapenem, and co-require impermeability to confer resistance in Enterobacteriaceae. Nevertheless, if ertapenem is to be used widely, its effects on the resistance ecology need to be monitored carefully.

Bacteria↗

In vitro effects of combinations of antipseudomonal agents against seven strains of multidrug-resistant Pseudomonas aeruginosa.

OBJECTIVES: The aim of this study was to evaluate the combined effects of antibiotic combinations by agar incorporation inhibitory tests and by time-kill tests on seven geographically and epidemiologically distinct isolates of multidrug-resistant Pseudomonas aeruginosa. All seven strains were resistant to piperacillin, meropenem, ceftazidime, cefoperazone-sulbactam, aztreonam, amikacin and ciprofloxacin. METHODS: Strains were distinguished by pulsed-field gel electrophoresis after DNA extraction and restriction with SpeI. MICs of the seven antibiotics listed above were determined by agar dilution. The effect of combinations of these agents was determined by agar incorporation tests and by time-kill studies. RESULTS: Among the two-drug combinations, the combination aztreonam and amikacin was the most effective, inhibiting proliferation in five of the seven strains. Among the three-drug combinations, the combinations of piperacillin, ceftazidime and amikacin, and that of ceftazidime, aztreonam and amikacin were the most effective, inhibiting proliferation in all seven strains. In the killing tests, the three-drug combination of ceftazidime, aztreonam and amikacin was the most effective. This three-drug combination had bacteriostatic effects on all seven strains 2, 4, 6 and 24 h after drug addition, synergic effects on 2-3 strains and bactericidal effects on 1-2 strains after 4, 6 and 24 h. CONCLUSIONS: The three-drug combination of ceftazidime, aztreonam and amikacin may be effective against P. aeruginosa resistant to all commonly used antipseudomonal drugs, and deserves further study.

Anti-Bacterial Agents↗

Reliability of routine disc susceptibility testing by the British Society for Antimicrobial Chemotherapy (BSAC) method.

OBJECTIVES: To ascertain the agreement between MICs determined at a central laboratory, and susceptible, intermediate and resistant categorizations based on zone diameters recorded at diagnostic laboratories using the BSAC standardized method. METHODS: Standardized disc susceptibility tests were performed at sentinel laboratories in three surveys, with MIC tests performed on the collected isolates at a reference laboratory. The organisms comprised over 3300 Enterobacteriaceae, Acinetobacter spp., pseudomonads, staphylococci and enterococci, with over 29 000 antibiotic/organism tests in total. RESULTS: More than 90% of the antibiotic/organism combinations classed as susceptible by disc tests in the sentinel laboratories were confirmed by MIC testing. Disagreements were more frequent where disc tests indicated resistance, with half of the piperacillin/tazobactam resistance and one-third of the cephalosporin resistance found in Enterobacteriaceae by disc tests not being confirmed, and with three-quarters of teicoplanin resistance in enterococci not confirmed. None of the few apparent cases of meropenem resistance in Enterobacteriaceae or linezolid, quinupristin/dalfopristin or vancomycin resistance in staphylococci were confirmed by MIC testing. When disagreements were found between disc- and MIC-based categorization, MICs were commonly, although not invariably, one to three doubling dilutions above or below the breakpoint. However, many of the disagreements where MICs were three or more dilutions from the breakpoint were not seen when disc tests were repeated in the central laboratory. CONCLUSIONS: The BSAC disc method seems adequate for confirming susceptibility to guide therapy and to monitor resistance trends. Nevertheless, there must be concern about the over-estimation of many resistances, and frequent zone:MIC disagreements for isolates with borderline susceptibility.

Bacteria↗

Selectivity of ertapenem for Pseudomonas aeruginosa mutants cross-resistant to other carbapenems.

OBJECTIVES: Ertapenem and other carbapenems will be used increasingly, as extended-spectrum beta-lactamases become more prevalent even among community-acquired pathogens. There is, however, concern that this use will select for resistances to imipenem and meropenem in nosocomial pathogens, notably Pseudomonas aeruginosa, and we investigated the validity of this concern. METHODS: Single-step selection experiments were performed by plating P. aeruginosa cultures on to agar containing doubling dilutions of ertapenem. MIC patterns, outer membrane protein profiles and the effects of efflux inhibitors were examined for selected mutants. RESULTS: At 2-8 x MIC, ertapenem selected (i) for OprD(-) mutants of P. aeruginosa, with cross-resistance only to carbapenems, (ii) for putative efflux types with broader cross-resistance, and also (iii) for various less familiar phenotypes. Efflux mutants were predominantly, but not exclusively, selected from carbenicillin-hypersusceptible strains and OprD(-) mutants largely from strains with normal levels of carbenicillin susceptibility. Whilst these data indicate potential cross-selectivity, they must be set against the observation that 20% serum raised the ertapenem MICs, and the drug concentrations needed for mutant selection, by over four-fold, reflecting the compound's strong protein binding. Since, following a 1 g intravenous dose the free ertapenem concentration in the serum falls below 4 mg/L--corresponding to the lower of two MIC(50) estimates--within 4 h (17% of the dosage interval) selectivity in vivo should be minimized. CONCLUSIONS: Whilst ertapenem can select for P. aeruginosa mutants with cross-resistance to imipenem and ertapenem in vitro, this selectivity should be minimal under clinical conditions.

Carbapenems↗

In vitro susceptibilities of aerobic and facultative Gram-negative bacilli isolated from patients with intra-abdominal infections worldwide: the 2003 Study for Monitoring Antimicrobial Resistance Trends (SMART).

OBJECTIVES: The SMART (Study for Monitoring Antimicrobial Resistance Trends) surveillance programme was begun in 2002 to monitor antimicrobial resistance trends among aerobic and facultative Gram-negative bacilli (GNB) isolated from intra-abdominal infections worldwide. METHODS: In 2003, 74 medical centres from 23 countries collected isolates for testing. Antimicrobial susceptibility testing was performed using broth microdilution according to the NCCLS guidelines for MIC testing. RESULTS: A total of 5658 aerobic and facultative GNB were isolated from intra-abdominal infections. Enterobacteriaceae composed 84% of the total isolates. Among the agents tested, the carbapenems were the most consistently active against the Enterobacteriaceae. E. coli was the most common isolate (46%), and the susceptibility rate to the quinolone (70-90% susceptible), cephalosporin (80-97% susceptible), aminoglycoside (77-100% susceptible) and carbapenem (99-100% susceptible) agents tested varied among geographic regions, with isolates from the Asia/Pacific region generally being the most resistant. Extended-spectrum beta-lactamases (ESBLs) were detected phenotypically in 9% of E. coli, 14% of Klebsiella spp., and 14% of Enterobacter spp. worldwide. ESBL producers generally had a more antibiotic-resistant profile than non-ESBL producers. CONCLUSIONS: Antimicrobial resistance among GNB isolated from intra-abdominal infections is a problem worldwide, especially in the Asia/Pacific region. The carbapenems ertapenem, meropenem and imipenem are highly active in vitro against Enterobacteriaceae isolated from intra-abdominal sites, including organisms that produce ESBLs.

Abdomen↗

Prevalence of Ambler class A and D beta-lactamases among clinical isolates of Pseudomonas aeruginosa in Korea.

OBJECTIVES: Recently, resistance to extended-spectrum cephalosporins due to acquired beta-lactamases has been reported in Pseudomonas aeruginosa. The aim of this study was to investigate the prevalence of Ambler class A and D beta-lactamases and their extended-spectrum derivatives and antimicrobial susceptibilities of P. aeruginosa isolated from various parts of Korea. METHODS: A total of 252 consecutive, non-duplicate isolates of P. aeruginosa were studied for the presence of class A or D beta-lactamase. Antibiotic susceptibility tests and PCR amplification of genes encoding class A (bla(PSE-1), bla(PER-1), bla(VEB-1), bla(TEM), bla(SHV), bla(CTX-M) and bla(GES-1)) and class D beta-lactamases (bla(OXA-groupI), bla(OXA-groupII) and bla(OXA-groupIII)) were performed. For PCR-positive isolates, isoelectric focusing (IEF) analysis, sequencing and pulsed-field gel electrophoresis (PFGE) were performed. RESULTS: In 64 (25.4%) isolates, structural genes for PSE-1 (6.3%), OXA-10 (13.1%), OXA-4 (4.3%), OXA-30 (2.0%), OXA-2 (2.3%) and OXA-17 (0.4%) were found; their distribution varied between provinces. None harboured bla(PER-1), bla(VEB-1), bla(TEM), bla(SHV), bla(CTX-M) and bla(GES-1). The cross-class resistance rates to other antibiotics was significantly higher in class A and D beta-lactamase producers than in non-producers (P < 0.001 for aminoglycosides, ciprofloxacin and meropenem). CONCLUSIONS: OXA-type beta-lactamases are widespread, but their extended-spectrum derivatives are rare among P. aeruginosa in Korea. To our knowledge, this is the first report of OXA-17, an extended-spectrum derivative of OXA-10, outside the Middle East. In addition, combined resistance to ticarcillin and aminoglycosides was a useful indicator for P. aeruginosa producing PSE- or OXA-type beta-lactamases in this study.

Drug Resistance, Bacterial↗

Antimicrobial susceptibility testing of Actinomyces species with 12 antimicrobial agents.

OBJECTIVE: This study was conducted to assess the susceptibility of human clinical isolates of Actinomyces species to 12 antimicrobial agents. METHODS: Human clinical isolates of Actinomyces spp. were collected from stored collections held at the Microbiology Department, Edinburgh University, Anaerobe Reference Laboratory, Cardiff, Glasgow Dental Hospital and Glasgow Royal Infirmary. Each isolate was identified by restriction analysis of amplified 16S ribosomal DNA. MICs of 12 antibiotics comprising benzyl penicillin, amoxicillin, ceftriaxone, linezolid, tetracycline, deoxycycline, clindamycin, erythromycin, clarithromycin, ciprofloxacin, meropenem and piperacillin/tazobactam for 87 strains of Actinomyces species were obtained by Etest methodology. RESULTS: The Actinomyces species identified for this study comprised: Actinomyces israelii, Actinomyces gerencseriae, Actinomyces turicensis, Actinomyces funkei, Actinomyces graevenitzii and Actinomyces europaeus. All isolates were susceptible to penicillin and amoxicillin. All but one strain of A. turicensis was susceptible to linezolid. A number of A. europaeus and A. graevenitzii isolates were resistant to ceftriaxone and piperacillin/tazobactam. A number of isolates of A. turicensis and A. europaeus also demonstrated resistance to erythromycin. All Actinomyces species tested appeared resistant to ciprofloxacin. CONCLUSIONS: Actinomyces species appear to be susceptible to a wide range of beta-lactam agents and these, when combined with beta-lactamase inhibitors, should be regarded as agents of first choice. Ciprofloxacin performed poorly. Tetracyclines also demonstrated poor performance. This is the first study of antimicrobial susceptibilities for a number of accurately identified clinical isolates of Actinomyces spp. There are a number of species differences in susceptibility profiles to the antimicrobials tested, suggesting that accurate identification and speciation may have an impact on clinical outcome.

Actinomyces↗