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Complementary activity of mezlocillin and the combination of amoxicillin with clavulanic acid on Enterobacteriaceae.

The MICs of amoxicillin, mezlocillin and BRL 25,000, a combination of two parts amoxicillin and one part clavulanic acid (2AM + 1CA), were measured for 331 Enterobacteriaceae strains which produced beta-lactamases as demonstrated by nitrocefin. The MIC values for mezlocillin and the combination 2AM + 1CA were very similar for the total number of the strains investigated. When investigated separately according to the bacterial species, three different sensitivity groups were established for the above-mentioned preparations: 1) species with the same or similar sensitivity to mezlocillin and 2AM + 1CA (Escherichia coli and Shigella spp., amoxicillin-resistant strains); 2) species which were more sensitive to mezlocillin than to the combination 2AM + 1CA (Citrobacter spp., Enterobacter cloacae, Serratia spp. and indole-positive Proteus as well as strains of E. coli and Shigella spp. which produce a cephalosporinase and are sensitive to amoxicillin); 3) species which are more sensitive to 2AM + 1CA than to mezlocillin (amoxicillin-resistant Salmonella spp., Proteus mirabilis and Klebsiella pneumoniae). This complementary activity of mezlocillin and 2AM + 1CA against Enterobacteriaceae depended on the beta-lactamases produced.

Amoxicillin↗

Evaluation of a semi-automated 24-hour commercial system for identification of Enterobacteriaceae and other gram-negative bacteria.

A semi-automated commercial system (ID 32 E, bioMérieux) for 24-hour identification of Enterobacteriaceae and other gram-negative fermentative and nonfermentative bacteria encountered in diagnostic microbiology was evaluated. Overall, the system correctly identified 506 (91.5%) of the 553 strains tested, 94 (17.0%) strains requiring additional tests for complete identification. Six (1.1%) strains were misidentified and 33 (6.0%) strains were not identified. Eight (1.4%) strains were not present in the database and were misidentified or not identified. The system is a suitable alternative to existing systems for the identification of Enterobacteriaceae and other gram-negative bacteria frequently encountered in clinical samples.

Bacterial Typing Techniques↗

Comparison of three methods for identification of Enterobacteriaceae.

This study compares the ability of three commercial overnight methods, API 20E, Minitek and Enteric-Tek, to accurately and completely identify 368 clinically isolated Enterobacteriaceae without supplemental tests. Organisms included Escherichia coli (54 strains), Shigella spp. (7), Edwardsiella tarda (1), Salmonella enteritidis (10), Citrobacter spp. (30), Klebsiella spp. (55), Enterobacter spp. (68), Hafnia alvei (2), Serratia spp. (33), Proteus spp. (64), Morganella morganii (24), Providencia spp. (18), and Yersinia enterocolitica (2). Methods were those of the manufactures without supplemental tests. API 20E correctly identified 93.2% of strains to species and 3.3% to genus level only, with 3.0% as part of a spectrum of identifications, and 0.5% incorrect identifications. Minitek yielded 96.0% correct identifications to species and 0.5% to genus level only, with 2.5% spectrum identifications, and 1.0% incorrect identifications. Enteric-Tek correctly identified 97.0% of strains to species level with 3.0% spectrum identifications. API 20E identification of some Serratia and Citrobacter strains was to genus level only. Problem organisms for Minitek included Enterobacter agglomerans and Serratia marcescens. A comparison of these three commercial methods shows that all three have the ability to identify most clinically isolated Enterobacteriaceae without supplemental tests.

Bacteriological Techniques↗

Performance of two four-hour identification systems with atypical strains of Enterobacteriaceae.

Two four-hour systems (Rapid 20E and Micro-ID) for identification of Enterobacteriaceae were evaluated using 66 strains isolated from clinical specimens which could not be identified by conventional methods. API 20E was used as reference method. Both systems had an acceptable identification rate (84.8% for Rapid 20E and 74.2% for Micro-ID). After referral to API's computer facilities the identification rate of Rapid 20E rose to 92.4%. Micro-ID incorrectly identified 16.7% of the strains and Rapid 20E only 1.5%. On the other hand, no identification was achieved in 13.6% of the strains using Rapid 20E versus 6% using Micro-ID. On the basis of these findings Rapid 20E is considered the more suitable system for the rapid identification of Enterobacteriaceae strains which cannot be identified by conventional methods.

Bacteriological Techniques↗

Cross-resistance of nalidixic acid resistant Enterobacteriaceae to new quinolones and other antimicrobials.

One hundred urine isolates Enterobacteriaceae screened for resistance to 30 micrograms nalidixic acid by disc diffusion test were examined by MIC determination for in vitro susceptibility to nalidixic acid, ciprofloxacin, enoxacin, gentamicin, nitrofurantoin, trimethoprim, cephalexin and ceftazidime. Those resistant to nalidixic acid and also gentamicin or a cephalosporin were further examined to determine the mechanism of resistance. Compared to the total urine isolates of Enterobacteriaceae from the same time period, this population as a whole was less susceptible to all antimicrobials tested except gentamicin. Strains that exhibited multiple resistance had the conventional mechanisms of resistance to those antimicrobials. No multiply resistant strains had a permeability barrier due to outer membrane protein alterations causing cross-resistance to chemically unrelated classes of antimicrobials.

Anti-Bacterial Agents↗

Occurrence of endemic plasmids causing beta-lactam resistance in Enterobacteriaceae in children's university hospital in Munich.

Susceptibility of 62 clinical isolates of Enterobacteriaceae to 15 aminoglycosides, beta-lactams and fluoroquinolones was determined. The isolates originating from 3 intensive care units (neonatal, pediatric, and surgical) and the Department of Infant Internal Medicine of the Children's University Hospital City Center in Munich (Germany) were collected in August 1999, and March and October 2000. Transferability of antibiotic resistance from donors to their E. coli transconjugants was also demonstrated. The majority of isolates were resistant to ampicillin, cefoxitin, ceftriaxone, cefotaxime, ceftazidime and azthreonam but they were susceptible to cefepime, meropenem, aminoglycosides and fluoroquinolones. The occurrence of beta-lactamases and extended-spectrum beta-lactamases (ESBL) was also shown. In August 1999 75% of isolates produced beta-lactamases and 15% ESBL, in March 2000 95% of isolates produced beta-lactamases and 9% ESBL; in October 2000 all isolates produced beta-lactamases and only 5% produced ESBL. Plasmid DNA analysis in randomly chosen isolates and their transconjugants revealed the presence of plasmids ranging from 19 to 136 kb; in the majority of isolates a 120-kb plasmid was observed. Further analysis using restriction endonuclease suggested a dissemination and persistence of an endemic plasmid at all 4 wards of the large pediatric hospital in the City Center of Munich which may be responsible for resistance to beta-lactams among Enterobacteriaceae isolates.

Child↗

Occurrence of strains producing specific antibacterial inhibitory agents in five genera of Enterobacteriaceae.

Striking differences in the production of specific inhibitory agents affecting other strains of the same (or of related) species were found between genera of the family Enterobacteriaceae. We tested 50-163 strains each of the potentially pathogenic genera: Escherichia, Citrobacter, Enterobacter, Kluyvera, and Leclercia for their ability to produce bacteriophages, high-molecular-weight (HMW) and low-molecular-weight (LMW) bacteriocins and siderophores against the same sets of strains, using the cross-test method. The genus Escherichia differs substantially from all other Enterobacteriaceae, harboring a notable proportion of lysogenic (36.6%) and colicinogenic (13.9%) strains. Only 18.2% of the Citrobacter strains are lysogenic and only rarely are they colicinogenic, although in 7.3%, they produce phage tail-like bacteriocins. On the other hand, Kluyvera strains were only in 1.8% lysogenic, no colicinogenic strains were found, but in 7.3%, they produced siderophores causing zones of growth inhibition in agar cultures of strains of the same genus. In Leclercia, 10.0% of the strains were lysogenic, 2.0% produced HMW bacteriocins, no colicinogenic strains were found and 2.0% produced siderophores. Enterobacter has shown 23.1% of strains producing siderophores, whereas merely 7.7% were lysogenic, 1.9% colicinogenic and 3.8% formed phage tail-like bacteriocins. HMW bacteriocins of Enterobacter strains disposed of an unusually wide spectrum of activity. The siderophore activity spectrum was rather wide in any genus, but the siderophores were usually not produced by strains producing phages or colicins.

Anti-Bacterial Agents↗

Antimicrobial susceptibility of respiratory isolates of Enterobacteriaceae and Staphylococcus aureus in Italy: incidence and trends over the period 1997-1999.

The antibiotic susceptibility of members of the family Enterobacteriaceae and of Staphylococcus aureus strains isolated from the respiratory tract was assessed over the period 1997-1999 as part of the Italian Epidemiological Observatory survey sponsored by the Smith-Kline Foundation. A standardised method was used to determine the MICs of 22 antibiotics against isolates of Klebsiella pneumoniae (n=870), Escherichia coli (n=684), Enterobacter cloacae (n=342), Enterobacter aerogenes (n=187) and Serratia marcescens (n=135) as well as the MICs of 11 antibiotics against isolates of Staphylococcus aureus (n=1,606). Overall, the susceptibility rate of Enterobacteriaceae isolates was > or = 90% to 5 agents (meropenem, imipenem, amikacin, cefepime and gentamicin); 89-80% to 2 agents (ciprofloxacin and tobramycin); and <80% to 11 agents (cefotaxime, piperacillin-tazobactam, trimethoprim-sulfamethoxazole, cefetamet, ceftriaxone, ceftazidime, aztreonam, ticarcillin-clavulanate, tetracycline, piperacillin, cefuroxime, chloramphenicol, ticarcillin, amoxicillin-clavulanate and amoxicillin). During the 3-year monitoring period, antibiotic susceptibility increased in Klebsiella pneumoniae against amoxicillin-clavulanate, in Escherichia coli against third-generation cephalosporins and aztreonam, in Enterobacter aerogenes against amoxicillin and piperacillin-tazobactam and in Serratia marcescens against most of the antibiotics. In contrast, Enterobacter cloacae showed a tendency to develop resistance to cefetamet, amikacin and ciprofloxacin. Of the total number of Staphylococcus aureus strains, 38% were methicillin resistant. Nearly 80% of the methicillin-resistant strains displayed a multiresistance pattern (additional resistance to 2 or more non-beta-lactam antibiotics). Rates of susceptibility of particular species (Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus) were compared using strains from different geographical areas of Italy (northern, central and southern) and from different nosocomial areas (outpatients, intensive care unit [ICU] inpatients, non-ICU inpatients). Susceptibility of Klebsiella pneumoniae to several antibiotics was lower in southern Italy, whereas the incidence of methicillin-resistant strains was higher in northern and central Italy. The susceptibility of Escherichia coli was similar in all three areas. No significant differences in susceptibility of Klebsiella pneumoniae or Escherichia coli were found between strains from inpatients and outpatients or from inpatients admitted to ICU and non-ICU units. The incidence of methicillin-resistant Staphylococcus aureus was higher in ICU inpatients (52%) than in non-ICU inpatients (38%) and lower in outpatients (19%) than in inpatients.

Drug Resistance, Bacterial↗

Psychrotrophic, proteolytic and lipolytic properties of Enterobacteriaceae isolated from milk and dairy products.

Most of the Enterobacteriaceae strains (73 out of 75) isolated in a previous study (Wessels et al., 1988) were psychrotrophic on agar plates, with the exception of Enterobacter cloacae strains. The Enterobacteriaceae strains were largely non-proteolytic on milk agar medium although limited numbers of E. cloacae, Serratia rubidaea and Klebsiella oxytoca strains were capable of proteolytic activity at 25 degrees C. The E. cloacae and K. oxytoca strains positive at 25 degrees C were also proteolytic at 7 degrees C. Most of the species tested were non-lipolytic on Victoria blue butterfat agar. The majority of Serratia marcescens and Klebsiella pneumoniae strains and a minority of E. cloacae and K. oxytoca strains, however, were lipolytic on this medium.

Animals↗

A new plate medium for rapid presumptive identification and differentiation of Enterobacteriaceae.

A new selective differential agar medium for rapid presumptive identification of Enterobacteriaceae from water and food samples is described (EMX ID agar). By a combination of fluorogenic and chromogenic substrates, the medium detects the presence of beta-D-glucuronidase, beta-D-galactosidase, beta-D-xylosidase, tryptophane deaminase and H2S; additionally, cytochrome-oxidase and indole production can be demonstrated. This medium provides an inexpensive means for simple and rapid presumptive identification of E. coli and coliforms and for the differentiation within the Klebsiella-Enterobacter and the Proteus-Providencia-Morganella group. Furthermore, it allows to distinguish between the H2S-positive Enterobacteriaceae Citrobacter freundii, Salmonella spp., S. arizonae, Edwardsiella, Proteus mirabilis, P. vulgaris and some oxidase-positive bacteria.

Agar↗

Development of resistance to aminoglycosides among coagulase-negative staphylococci and enterobacteriaceae in a neonatal intensive care unit.

A prospective study of the development of resistance to aminoglycosides among coagulase-negative staphylococci (CNS) and Enterobacteriaceae (ENT) was conducted for all patients admitted to a neonatal intensive care unit (NICU) from October 1985 to January 1990. A change in antibiotic regimen from gentamicin to amikacin occurred in January 1986, due to widespread gentamicin resistance among CNS, the most important cause of nosocomial infections in this NICU. From 657 patients, 884 faecal cultures, 1505 cultures from the respiratory tract and 152 blood cultures were included in the study. After its introduction, susceptibility to amikacin decreased rapidly in faecal and respiratory CNS isolates (from 62% to 28% and from 58% to 23% respectively). During the first half year, resistance to amikacin in faecal CNS isolates developed more rapidly among antibiotic-treated patients than among patients not treated with antibiotics. Susceptibility to amikacin in CNS blood isolates decreased more slowly, from 94% to 58% in 1987, while subsequently an increase in susceptibility was observed to about 80% in 1989. The same difference in development of resistance in faecal and respiratory CNS isolates compared to CNS blood isolates was noticed for gentamicin and tobramycin. In contrast, ENT remained highly (85-100%) susceptible to amikacin, gentamicin and tobramycin throughout the study period. It was concluded that four years after its introduction amikacin still appeared to be a valuable antibiotic in combination treatment of the vast majority of clinically important infections occurring in our NICU, since both Enterobacteriaceae and the majority of CNS blood isolates proved to be susceptible to this agent.(ABSTRACT TRUNCATED AT 250 WORDS)

Amikacin↗

Evaluation of in vitro antimicrobial activity of lomefloxacin against staphylococci, enterococci, Enterobacteriaceae, and Pseudomonas aeruginosa.

In vitro antimicrobial activity of lomefloxacin and other antibiotics (norfloxacin, beta-lactams, cotrimoxazole, netilmicin, and miokamycin) was evaluated against 317 clinical isolates including Staphylococcus aureus, enterococci, Enterobacteriaceae, and Pseudomonas aeruginosa. Lomefloxacin showed a high in vitro activity against a wide variety of bacterial species. Against Enterobacteriaceae, lomefloxacin displayed the highest activity (MIC90S, less than or equal to 1 microgram/ml), and it was usually more active than ampicillin and netilmicin, and often more active than cotrimoxazole. Lomefloxacin showed a good antimicrobial activity also against both methicillin-susceptible and methicillin-resistant S. aureus, as it was able to inhibit 90% of staphylococcal strains at a concentration less than or equal to 2 micrograms/ml. Against enterococci and P. aeruginosa, lomefloxacin was usually less active (MIC90S, 8 micrograms/ml). As compared to norfloxacin, lomefloxacin displayed an overall similar activity against staphylococci and enterococci, but appeared less active against Gram-negative bacteria. Bactericidal activity of lomefloxacin against E. coli, when assayed by a novel method, proved to be high, and results indicate that an oral dose of 200 mg of lomefloxacin should exert a very high bactericidal activity against E. coli in urinary tract infections, even in those sustained by large bacterial populations.

4-Quinolones↗

In vitro activity of ampicillin, amoxicillin, ampicillin-sulbactam, and amoxicillin-clavulanic acid against consecutive clinical isolates of Enterobacteriaceae.

Ampicillin was generally twice as active as amoxicillin against 2440 consecutive isolates of Enterobacteriaceae from five medical centers. When beta-lactamase inhibitors were added to the penicillins, there was a significant increase in susceptibility. The magnitude of the increased susceptibility to ampicillin-sulbactam (A-S) and amoxicillin-clavulanic (A-C) acid varied with the species and types of beta-lactamases elaborated. Although cross-susceptibility and cross-resistance between ampicillin and amoxicillin was nearly complete, major differences were documented between A-S and A-C with 6.7% of our consecutive isolates of Enterobacteriaceae. The clinical significance of these findings remains uncertain, but they may help explain some of the discrepancies occasionally observed by clinical microbiologists with the combination drugs.

Amoxicillin↗

Shedding of antibiotic-resistant members of the family Enterobacteriaceae in healthy residents of France and Jordan.

We compared the frequency of shedding of members of the family Enterobacteriaceae resistant to ampicillin, tetracycline, chloramphenicol, kanamycin, gentamicin and ceftazidime in 83 French residents of the Paris urban area and in 101 subjects in Jordan, 64 of whom resided in the urban area of Irbid, 15 in rural areas, and 22 of whom had a nomadic lifestyle. There was no significant difference between these populations regarding (i) the percentages of subjects with strains resistant to any of the antimicrobial agents tested and (ii) the proportions of total counts of organisms of the Enterobacteriaceae resistant to these agents. The simultaneous shedding of strains resistant to ampicillin, chloramphenicol, tetracycline and kanamycin was significantly associated with (i) exposure to antibiotic treatment during the six months preceding the study and (ii) the presence of many children at home.

Adult↗

Taxonomic diversity of anaerobic glycerol dissimilation in the Enterobacteriaceae.

A total of 1,123 strains representing 128 taxa in the Enterobacteriaceae (named species or subspecies and genomic species) were screened for the presence of glycerol dehydrogenases and 1,3-propanediol dehydrogenase. Only eight taxa, Citrobacter freundii sensu stricto, C. youngae, C. braakii, C. werkmanii, Citrobacter genomospecies 10 and 11, Enterobacter gergoviae and Klebsiella pneumoniae subsp. pneumoniae could grow fermentatively on glycerol and possessed both glycerol dehydrogenase type I (induced by glycerol and dihydroxyacetone) and 1,3-propanediol dehydrogenase which are typical enzymes of the anaerobic glycerol dissimilation pathway. Six other species, C. koseri, E. aerogenes, E. intermedium, K. oxytoca, K. planticola and K. terrigena could not grow fermentatively on glycerol and possessed a glycerol dehydrogenase type I but no 1,3-propanediol dehydrogenase. Other glycerol dehydrogenases types were found: type II (induced by glycerol and hydroxyacetone), type III (induced by glycerol only) and type IV (induced by hydroxyacetone only). They were widely distributed among the Enterobacteriaceae. Classification and identification may take advantage of tests exploring the dissimilation of glycerol.

Enterobacteriaceae↗

Increasing trend in the prevalence of plasmid-mediated AmpC beta-lactamases in Enterobacteriaceae lacking chromosomal ampC gene at a Korean university hospital from 2002 to 2004.

The aim of the study is to investigate the prevalence of plasmid-mediated AmpC beta-lactamases in Enterobacteriaceae naturally lacking chromosomal AmpC beta-lactamases. A total of 1860 clinical isolates of Klebsiella spp., Salmonella spp., and Proteus mirabilis were collected from a Korean hospital between January 2002 and December 2004. For the isolates that are nonsusceptible to cefoxitin, polymerase chain reaction amplification of the bla(SHV), bla(TEM), and bla(AmpC) genes and sequencing were performed. Plasmid-mediated AmpC beta-lactamases were found in 2.9% (37 isolates of DHA-1, 1 isolate of CMY-1, 1 isolate of CMY-2, and 1 isolate of ACT-1) of Klebsiella pneumoniae, 2.5% (5 isolates of DHA-1) of Klebsiella oxytoca, 0.8% (1 isolate of DHA-1) of Salmonella spp., and none of P. mirabilis isolates. The DHA-1-producing K. pneumoniae was only 2 isolates (0.6%) in 2002, but the rate and the number significantly increased to 2.4% (13 of 538 isolates) in 2003 and to 4.3% (22 of 512) in 2004. In conclusion, DHA-1 is the most prevalent plasmid-mediated AmpC beta-lactamase in Enterobacteriaceae lacking chromosomal ampC gene, and the DHA-1-producing K. pneumoniae isolates have rapidly increased since 2003 in a Korean hospital. In addition, this is the first report of the appearance of a K. pneumoniae isolate producing ACT-1 beta-lactamase in Korea.

Bacterial Proteins↗

Comparative in vitro activity and the inoculum effect of ertapenem against Enterobacteriaceae resistant to extended-spectrum cephalosporins.

The in vitro activity and the inoculum effect of ertapenem were evaluated against a total of 70 Enterobacteriaceae isolates resistant to extended-spectrum cephalosporins. The extended-spectrum beta-lactamase phenotypic confirmatory disk diffusion test was performed and AmpC-inducible species were detected using cefoxitin/cefotaxime disk antagonism tests. beta-Lactamases were characterised by isoelectric focusing and TEM-specific polymerase chain reaction. Minimum inhibitory concentrations (MICs) were determined by the National Committee for Clinical Laboratory Standards agar dilution method. Ertapenem showed excellent activity against almost all isolates tested, with MIC(50) and MIC(90) values of 0.03 mg/L and 0.12 mg/L, respectively. When the inoculum was increased 100-fold, susceptibility decreased from 98.6% to 65.7% for cefepime and from 75.7% to 54.3% for piperacillin/tazobactam, without changing for ertapenem. The data from this study suggest that this new carbapenem may be useful for treating mixed infections involving Enterobacteriaceae isolates resistant to third-generation cephalosporins.

Anti-Bacterial Agents↗

The molecular and clinical epidemiology of enterobacteriaceae-producing extended-spectrum beta-lactamase in a tertiary care hospital.

To describe the epidemiology of Enterobacteriaceae-producing extended-spectrum beta-lactamase (EP-ESBL) in a non-outbreak setting, and to define the risk factors associated with colonization, a 5-month surveillance study was initiated. Ten of 333 patients were colonized with EP-ESBL, as defined by isoelectric focusing. Klebsiella sp. and Escherichia coli were the species most commonly harbouring these plasmid-mediated enzymes. Of the 16 SHV-producing isolates, 10 were SHV-3-like (pI 7.0) and six were SHV-5-like (pI 8.2). All isolates were resistant to ceftriaxone. Ceftazidime resistance was detected in 50% and 100% of SHV-3-like and SHV-5-like producing isolates, respectively. One patient was colonized with four different SHV-5-like producing Enterobacteriaceae. These isolates carried plasmids that were indistinguishable by restriction endonuclease analysis, indicating broad plasmid transfer within the patient. By logistic regression, haemodialysis was a strong risk factor for colonization with EP-ESBL, suggesting that, in our hospital, horizontal transmission is an important mechanism of dissemination of these resistant pathogens.

Academic Medical Centers↗