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PubMed1998

Trends in beta-lactam resistance among Enterobacteriaceae.

beta-Lactam resistance among Enterobacteriaceae is related primarily to the emergence of novel beta-lactamases. The class A extended-spectrum beta-lactamases hydrolyze extended-spectrum beta-lactams and are inhibited by clavulanic acid. These beta-lactamases are divided in two groups: TEM and SHV derivatives and non-TEM and non-SHV extended-spectrum beta-lactamases (CTX-M1, CTX-M2, MEN-1, PER-1, PER-2, TOHO-1, and VEB-1). The plasmid-mediated cephalosporinases (MIR-1, FOX-1, MOX-1, BIL-1, CMY-1, CMY-2, and LAT-1) hydrolyze extended-spectrum cephalosporins and cephamycins and are not inhibited by clavulanic acid. They have been reported in Europe and in the United States. The 15 inhibitor-resistant penicillinases are TEM derivatives (except for SHV-10) and plasmid mediated, and they are mainly from Escherichia coli isolates. The carbapenemases noted among Enterobacteriaceae are either the chromosomally located penicillinases (Sme-1, NmcA, IMI-1) found in rare Enterobacter cloacae or Serratia marcescens isolates or the plasmid-mediated metalloenzyme IMP-1 that is widespread in Japan. The incidence of resistance among Enterobacteriaceae related to the other more common beta-lactam-resistance mechanisms has continued to rise worldwide.

Animals↗
PubMed1997

Contrasting effects of lipopolysaccharides (endotoxins) from oral black-pigmented bacteria and Enterobacteriaceae on platelets, a major source of serotonin, and on histamine-forming enzyme in mice.

By measurement of serotonin levels, the translocation of platelets to various tissues was examined following intravenous injection of a lipopolysaccharide (LPS) into C3H/HeN mice. There was a rapid platelet accumulation (within 5 min and particularly in the lung), followed by a slower accumulation in the liver, which reached its plateau 3-5 h later. The severity of the anaphylactoid shock corresponded well with the magnitude of the rapid response. LPSs from the oral black-pigmented bacteria, Porphyromonas gingivalis and Prevotella intermedia, were much more potent in inducing the rapid platelet response than were those from the Enterobacteriaceae Escherichia coli and Salmonella typhimurium. However, LPSs from these Enterobacteriaceae were significantly more potent than those from black-pigmented bacteria in inducing the slow platelet response. There was also a contrast between their abilities to induce histidine decarboxylase, which forms histamine from histidine: LPSs from the Enterobacteriaceae were much more potent than those from black-pigmented bacteria.

Acetylmuramyl-Alanyl-Isoglutamine↗
PubMedDate not supplied

Epidemiology of infections caused by gentamicin-resistant enterobacteriaceae and Pseudomonas aeruginosa over 15 years at the Nashville Veterans Administration Medical Center.

Nosocomial infections and gentamicin resistance were surveyed over 15 years at Nashville Veterans Administration Medical Center, and trends for Enterobacteriaceae and Pseudomonas aeruginosa were contrasted. Analysis of approximately 6,000 nosocomial infections indicated that four-fifths were caused by aerobic gram-negative bacilli. Three hospital-wide outbreaks caused by Enterobacteriaceae occurred; these three outbreaks were due to Serratia marcescens, Klebsiella pneumoniae, and Enterobacter cloacae, respectively. The outbreaks were temporally related to the emergence of gentamicin resistance. Detailed analysis of the recent outbreak due to Enterobacter indicated that an increasing prevalence of gentamicin-resistant E. cloacae predated nosocomial infections by several months; this pattern suggested that such outbreaks could be predicted. Molecular epidemiologic data pertaining to the preservation over a decade of genes encoding gentamicin resistance were reviewed. In contrast to Enterobacteriaceae, P. aeruginosa gradually and progressively developed resistance to gentamicin that spread in an endemic fashion, with parallel increases in nosocomial infections. This pattern appeared to relate to different modes of spread and persistence for resistant P. aeruginosa that may require unique methods for control.

Cross Infection↗
PubMed1983

Differential susceptibility of Aeromonas hydrophila and Enterobacteriaceae to nalidixic acid.

The susceptibility of 49 strains of Aeromonas hydrophila and 77 strains of Enterobacteriaceae towards nalidixic acid was compared by tube dilution and agar diffusion methods. A higher susceptibility was exhibited by Aerom. hydrophila and no overlap in MICs was found for strains of the two groups of microorganisms. Discs containing 0.25 microgram of nalidixic acid produced measurable zones of growth inhibition with Aerom. hydrophila and no zones with the Enterobacteriaceae. The use of a disc with 0.25 microgram of nalidixic acid for the primary differentiation of strains of Aerom. hydrophila from those of the Enterobacteriaceae with similar biochemical properties is suggested.

Aeromonas↗
PubMed1987

The in-vitro activity of metronidazole against Enterobacteriaceae alone and in mixed cultures with Bacteroides fragilis.

The in-vitro antimicrobial action of therapeutic concentrations of metronidazole against Bacteroides fragilis and six different strains of Enterobacteriaceae in pure and mixed cultures have been studied. Under anaerobic conditions, metronidazole suppressed the growth of pure cultures of the Enterobacteriaceae. A reduction in the viable counts from 10(9) cfu/ml, in the 24 h controls, to 10(8), 10(7) and 10(5) cfu/ml in the presence of 10, 50 and 100 mg/l of metronidazole respectively, was observed. These concentrations of drug produced a marked bactericidal effect against B. fragilis, as expected. The antimicrobial activity of metronidazole on mixed cultures of B. fragilis and each one of the Enterobacteriaceae studied was greater against both micro-organisms than the corresponding effect on their respective pure cultures, under the same experimental conditions.

Bacteroides fragilis↗
PubMed1994

Use of cefotaxime or ceftazidime susceptibility tests for predicting susceptibility of Enterobacteriaceae and Pseudomonas aeruginosa.

To determine if susceptibility to aztreonam could be predicted from cefotaxime or ceftazidime disc diffusion testing, 919 Enterobacteriaceae and 187 Pseudomonas aeruginosa clinical strains were studied. The correlation coefficient between the diameters of inhibition zones was 0.9 for cefotaxime versus aztreonam and ceftazidime versus aztreonam comparisons in Enterobacteriaceae and 0.75 for ceftazidime versus aztreonam comparison in P. aeruginosa. For 99% of the Enterobacteriaceae, there was no risk in predicting susceptibility to aztreonam on the basis of cefotaxime or ceftazidime susceptibility tests. To minimize the risk of the remaining 1% of the strains being erroneously classified as susceptible to aztreonam, ceftoaxime should be tested in preference to ceftazidime, and the production of extended-spectrum beta-lactamases should be tested for using the cefotaxime-clavulanate disc synergy test. For P. aeruginosa strains, susceptibility to aztreonam could be accurately predicted from ceftazidime susceptibility tests for ticarcillin susceptible strains, but for ticarcillin resistant strains, susceptibility to aztreonam should be tested.

Aztreonam↗
PubMed1995

The comparative inhibitory and bactericidal activities of meropenem and imipenem against Acinetobacter spp. and Enterobacteriaceae resistant to second generation cephalosporins.

The inhibitory and bactericidal activities of meropenem and imipenem were compared using Acinetobacter spp. and Enterobacteriaceae resistant to second generation cephalosporins. The inhibitory activity of both agents was similar for Acinetobacter spp. but meropenem was more active against the Enterobacteriaceae. Bactericidal activities were compared employing time-kill curves which indicated meropenem was more bactericidal than imipenem against Enterobacteriaceae at pharmacologically achievable concentrations while imipenem was more bactericidal than meropenem against Acinetobacter spp.

Acinetobacter↗
PubMed1997

Prevalence and mechanism of resistance to 'third-generation' cephalosporins in clinically relevant isolates of Enterobacteriaceae from 43 hospitals in the UK, 1990-1991.

In a UK survey of the occurrence of extended spectrum beta-lactamases, 96 hospitals submitted a total of 3951 non-selected, non-duplicate isolates of Enterobacteriaceae from 100 patients in each hospital, 206 of these cultures being mixed and, therefore, discarded. These isolates were initially screened for strains likely to produce extended-spectrum beta-lactamases (ESBLs) by MIC determination of beta-lactams followed by a bioassay, then disc approximation test and isoelectric focusing (IEF). Isolates were further examined using two pairs of PCR primers for both blaTEM and blaSHV genes. The ability of isolates to transfer resistance to both cefotaxime and ceftazidime by conjugation and transformation were examined. Four hundred and nine cefotaxime/ceftazidime-resistant isolates (10.9%) were identified from the 3745 submitted isolates, of which 338 (9.0%) were Enterobacteriaceae, 29 Escherichia coli, 35 Klebsiella spp. and seven Hafnia alveii. IEF suggested that 17 isolates produced an ESBL, which was confirmed in most cases by PCR and hydrolysis, five isolates produced an SHV enzyme by IEF, but not confirmed by PCR, and 11 had isoelectric points in the range 8-9 suggesting a possible AmpC enzyme. Only two isolates transferred the determinants. In the case of the Klebsiella spp., 19 of the 24 ceftazidime-resistant/clavulanate-sensitive isolates were positive by PCR for a blaSHV gene. No isolates were identified as carrying blaTEM, although eight isolates had isoelectric points of 5-6.3, suggesting the presence of a possible TEM beta-lactamase. The results for the H. alveii isolates suggest that either an AmpC-like enzyme or a transferable beta-lactamase which is not TEM/SHV is present. This study shows that a wide range of genotypically and phenotypically different isolates of Enterobacteriaceae producing ESBL-like enzymes is present throughout the UK at a frequency of about 1% of unselected isolates. It is important that surveillance of resistance to these clinically important antibiotics is maintained as the occurrence of localized or more widespread outbreaks caused by bacteria producing ESBLs is to be expected.

Cephalosporins↗
PubMed2005

Emergence of Enterobacteriaceae producing extended-spectrum beta-lactamases (ESBLs) in the community.

Enterobacteriaceae, especially Klebsiella spp. producing extended-spectrum beta-lactamases (ESBLs) such as SHV and TEM types, have been established since the 1980s as a major cause of hospital-acquired infections. Appropriate infection control practices have largely prevented the dissemination of these bacteria within many hospitals, although outbreaks have been reported. However, during the late 1990s and 2000s, Enterobacteriaceae (mostly Escherichia coli) producing novel ESBLs, the CTX-M enzymes, have been identified predominantly from the community as a cause of urinary tract infections. Resistance to other classes of antibiotics, especially the fluoroquinolones, is often associated with ESBL-producing organisms. Many clinical laboratories are still not aware of the importance of screening for ESBL-producing Enterobacteriaceae originating from the community. A heightened awareness of these organisms by clinicians and enhanced testing by laboratories, including molecular surveillance studies, is required to reduce treatment failures, to limit their introduction into hospitals and to prevent the spread of these emerging pathogens within the community.

Community-Acquired Infections↗
PubMed1979

Antigenic cross-reactivity of major outer membrane proteins in enterobacteriaceae species.

The protein constituents in the outer membrane (OM) of several serotypes of Escherichia coli and some other Enterobacteriaceae cross-reacted antigenically. Solubilized OM preparations of these bacteria were applied in interfacial precipitin tests to antisera elicited in rabbits against whole bacterial cells, absorbed with their appropriate lipopolysaccharide before testing. The resulting immunecomplexes were analysed on polyacrylamide gels. Protein profiles of the immunoprecipitates showed a considerable antigenic cross-reactivity of outer membrane proteins between most E. coli serotypes. Cross-reactivity, though substantially lower, was also found with OM from three other Enterobacteriaceae species, but was not detectable with Pseudomonas aeruginosa OM. When OM preparations were solubilized at room temperature, the peptidoglycan-bound proteins in the molecular weight range 37,000 to 41,000 predominated in the protein profiles of the immunecomplexes. In profiles of immunecomplexes obtained with boiled OM preparations, a heat-modifiable protein (mol. wt 33,000) predominated. The major OM proteins of the Gram-negative bacterium may therefore play a role as common surface antigens of the family of Enterobacteriaceae.

Antigen-Antibody Complex↗
PubMed1980

Major outer membrane proteins: common antigens in enterobacteriaceae species.

The major outer membrane (OM) proteins of 23 enterobacterial strains (principally clinical isolates) and five non-Enterobacteriaceae species were investigated by the sodium dodecyl sulphate-polyacrylamide gel immunoperoxidase (SGIP) technique to evaluate antigenic cross-reactivity among these proteins. All enterobacterial strains contained one or more peptidoglycan-associated major OM proteins, cross-reactive with the peptidoglycan-bound protein I of Escherichia coli, and one non-peptidoglycan-bound heat-modifiable protein, cross-reactive with protein II of E. coli. Results indicated that antigenic cross-reactivity of the major OM proteins is a general phenomenon in the family Enterobacteriaceae, independent of any molecular weight variation of the corresponding proteins in different bacterial strains. SGIP experiments carried out with OM preparations of other species showed no cross-reactivity of any of their OM proteins with enterobacterial major OM proteins. The significance of the immunological relatedness of OM proteins for the classification of some Enterobacteriaceae is discussed.

Antigens, Bacterial↗
PubMed2006

Characterization of the Enterobacteriaceae isolated from an artisanal Italian ewe's cheese (Pecorino Abruzzese).

AIMS: To evaluate some physiological characteristics of the Enterobacteriaceae isolated from Pecorino cheese. METHODS AND RESULTS: The production of organic acids, secondary volatile compounds, biogenic amines (BA) and the lipolytic and proteolytic activities of Citrobacter braakii, Enterobacter sakazakii, Escherichia coli, Kluyvera spp., Salmonella enterica ssp. arizonae and Serratia odorifera strains were determined in skim milk after 48 h of fermentation at 30 degrees C. The proteolytic activity observed only in Ser. odorifera and Kluyvera spp. was confirmed by the peptide profiles of the pH 4.6-insoluble fraction using RP-HPLC; however, the lipase activity was evidenced in all the isolates of E. coli, Kluyvera spp. and Salm. enterica ssp. arizonae. During fermentation, all the strains utilized citric acid and produced significant quantities of putrescine followed by histamine, spermine and spermidine as well as acetic and lactic acid. Moreover, the major volatile compounds produced were ethanol, 2,3-butanedione, 3-hydroxy-2-butanone, 2-heptanone and acetone. CONCLUSIONS: The Enterobacteriaceae of dairy origin possess many metabolic activities that could affect the sensory quality of the cheese in which they grow during ripening. SIGNIFICANCE AND IMPACT OF THE STUDY: The important physiological characteristics possessed by Enterobacteriaceae confirm the complexity of the microbiota of Pecorino Abruzzese cheese, which influences the typical sensory properties of this product.

Acetic Acid↗
PubMed2007

Both sulfate-reducing bacteria and Enterobacteriaceae take part in marine biocorrosion of carbon steel.

AIMS: In order to evaluate the part played in biocorrosion by microbial groups other than sulfate-reducing bacteria (SRB), we characterized the phylogenetic diversity of a corrosive marine biofilm attached to a harbour pile structure as well as to carbon steel surfaces (coupons) immersed in seawater for increasing time periods (1 and 8 months). We thus experimentally checked corroding abilities of defined species mixtures. METHODS AND RESULTS: Microbial community analysis was performed using both traditional cultivation techniques and polymerase chain reaction cloning-sequencing of 16S rRNA genes. Community structure of biofilms developing with time on immersed coupons tended to reach after 8 months, a steady state similar to the one observed on a harbour pile structure. Phylogenetic affiliations of isolates and cloned 16S rRNA genes (rrs) indicated that native biofilms (developing after 1-month immersion) were mainly colonized by gamma-proteobacteria. Among these, Vibrio species were detected in majority with molecular methods while cultivation techniques revealed dominance of Enterobacteriaceae such as Citrobacter, Klebsiella and Proteus species. Conversely, in mature biofilms (8-month immersion and pile structure), SRB, and to a lesser extent, spirochaetes were dominant. CONCLUSIONS: Corroding activity detection assays confirmed that Enterobacteriaceae (members of the gamma-proteobacteria) were involved in biocorrosion of metallic material in marine conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: In marine biofilms, metal corrosion may be initiated by Enterobacteriaceae.

Base Sequence↗
PubMed2006

Comparison of workflow and accuracy of identification and antimicrobial susceptibility testing of clinical isolates of Enterobacteriaceae, Pseudomonas aeruginosa and enterococci by Vitek 2 and routine methods.

Three hundred and fifty-three consecutive urine cultures growing Enterobacteriaceae, Pseudomonas aeruginosa or enterococci were subjected to parallel identification (ID) and antimicrobial susceptibility testing (AST) by Vitek 2 and routine methods, including simple screening tests or API 20 E for ID and standardized disc diffusion for AST. Accuracy of results, technician hands-on time required by both methods and time to results were compared. Vitek 2 correctly identified 322 (94.7%) of the 340 gram-negative isolates and 17 (81%) of the 21 Enterococcus faecalis strains. AST by Vitek 2 and disc diffusion gave category agreement for 4,058 (95.5%) of 4,248 organism-antimicrobial agent combinations. With MIC determination by E-test as reference, AST by Vitek 2 and disc diffusion produced 15 and 3 very major errors, respectively. Six (40%) of the fifteen very major errors by Vitek 2 were associated with trimethoprim-sulfamethoxazole. With an average of 22 specimens processed per day, use of Vitek 2 saved 80 min per day of technician hands-on time as compared to routine methods. Regarding the cost of hands-on worktime and consumables, use of Vitek 2 for identification of Escherichia coli-screened Enterobacteriaceae saved 0.70 p per sample in comparison to API 20 E. More than 80% of Enterobacteriaceae introduced to Vitek 2 in the morning could be reported by 16:00.

Anti-Bacterial Agents↗
PubMed1984

Microtests for rapid identification of Enterobacteriaceae. Four-hour tests by use of substrate-impregnated paper discs.

A number of microtests for the diagnosis of Enterobacteriaceae in 4 hours were developed. Composition of substrates and preparation of reaction wells are described. There were used substrate-impregnated paper discs affixed to microwell plates. Test results of 253 stock cultures of Enterobacteriaceae were correlated with results obtained by conventional tube methods. Twenty out of 23 microtests showed total correlations of more than 95%, and the remaining 3 tests 85-94%. The microtests are concluded to be economic and reliable alternatives to conventional tube methods and seem suitable for routine identification of Enterobacteriaceae.

Bacteriological Techniques↗
PubMed1976

Comparative activity of netilmicin, gentamicin, amikacin, and tobramycin against Pseudomonas aeruginosa and Enterobacteriaceae.

Netilmicin (Sch 20569), a semisynthetic aminoglycoside antibiotic, was compared with gentamicin, tobramycin, and amikacin against 242 clinical isolates of Pseudomonas and Enterobacteriaceae. The minimum inhibitory concentration (MIC) was determined in both solid and liquid media. Netilmicin exhibited typical aminoglycoside properties, such as little effect of inoculum size on MIC, relatively small gap between MIC and minimum bactericidal concentration, and potentiation of anti-Pseudomonas activity in the presence of carbenicillin. Netilmicin provided no advantage in antimicrobial activity over gentamicin for either Pseudomonas or Enterobacteriaceae. Nearly complete cross-resistance to netilmicin was encountered with isolates resistant to gentamicin in either solid or liquid media. Netilmicin was less active than gentamicin against isolates of Pseudomonas and Providencia. Major discrepancies between MIC values determined in agar as opposed to those determined in broth were encountered for most isolates of Pseudomonas but also, depending upon antibiotic tested, for between 15 and 40% of isolates of Enterobacteriaceae. This new aminoglycoside agent will be useful clinically only if it is shown to be significantly less toxic than presently available analogues.

Amikacin↗
PubMed1987

Distribution of erythromycin esterase and rRNA methylase genes in members of the family Enterobacteriaceae highly resistant to erythromycin.

The distribution of nucleotide sequences related to ereA, ereB, and ermAM was studied by colony hybridization in 112 strains of members of the family Enterobacteriaceae that are highly resistant to erythromycin. The ereA and ereB genes encoding erythromycin esterases type I and II, respectively, were detected in strains inactivating the 14-membered macrolides erythromycin and oleandomycin. Because all 52 strains resisting these antibiotics by inactivation were detected by ereA (n = 23), ereB (n = 23), or both probes (n = 6), only two classes of genes accounted for this resistance phenotype. The ermAM gene encoding a streptococcal rRNA methylase was detected in 21 strains of Escherichia coli and two strains of Klebsiella spp. Determination of the MICs of macrolide, lincosamide, and streptogramin (MLS) antibiotics demonstrated a correlation between hybridization with ermAM and the so-called MLS resistance phenotype. The presence of 11 strains coresistant to MLS antibiotics that did not hybridize to the ermAM probe suggests that, as in gram-positive organisms, MLS resistance in members of the family Enterobacteriaceae involves more than one class of rRNA methylase. Numerous strains (n = 18) were found to produce both an erythromycin esterase type II and an rRNA methylase. Physical linkage between ereB and ermAM may be responsible for the codissemination of the genes. Despite their exogenous origin, ereB and ermAM are already disseminated in various genera of the Enterobacteriaceae.

Carboxylic Ester Hydrolases↗
PubMed1990

Beta-lactamase production in members of the family Enterobacteriaceae and resistance to beta-lactam-enzyme inhibitor combinations.

Recent reports that members of the family Enterobacteriaceae that produce high levels of certain beta-lactamases are often resistant to ticarcillin-clavulanate prompted this study to assess the relationship between type and amount of enzyme produced and susceptibility to ticarcillin-clavulanate, piperacillin-tazobactam, and cefoperazone-sulbactam. Agar dilution MICs were determined by using 73 strains of Enterobacteriaceae that produced a single beta-lactamase that had been characterized and quantified and a beta-lactamase-negative control strain of Escherichia coli. For E. coli and Klebsiella pneumoniae, MICs of each combination increased as levels of TEM, SHV-1, or class IV enzymes increased. However, the percentage of strains that were resistant was highest for ticarcillin-clavulanate (32%), with only 18 and 6% resistant to piperacillin-tazobactam and cefoperazone-sulbactam, respectively. Strains producing PSE-1, regardless of level, were resistant or moderately susceptible to ticarcillin-clavulanate but were susceptible to piperacillin-tazobactam and cefoperazone-sulbactam. HMS-1 and OHIO-1 beta-lactamases were associated with resistance to ticarcillin-clavulanate and piperacillin-tazobactam, respectively. High levels of class IV enzymes in Klebsiella oxytoca were associated with resistance to all three combinations. These results indicate that the level and type of beta-lactamase produced by members of the family Enterobacteriaceae are important determinants of susceptibility to beta-lactam-inhibitor combinations, especially ticarcillin-clavulanate.

Anti-Bacterial Agents↗