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PubMed2000

Variations in antibiotic resistance profile in Enterobacteriaceae isolated from wild Australian mammals.

We carried out a retrospective analysis of 946 strains of Enterobacteriaceae isolated from wild Australian mammals between 1993 and 1997. The prevalence of resistance to fixed concentrations of 32 antimicrobial agents was determined, and the respective roles that taxonomic family of the host, state of origin and bacterial species play in defining prevalence and range of resistance were investigated. Our results demonstrated a low but widespread prevalence of antimicrobial resistance in wild isolates. Only amikacin, ciprofloxacin, meropenem and gentamicin inhibited growth in all 946 samples. There was extensive variation in the combination of antibiotics to which isolates were resistant, and multiple antibiotic resistance was common. Geographical location and host group significantly influenced the antibiotic resistance profile of an isolate, whereas bacterial species influenced both the resistance profile of an isolate and the number of antibiotics it was resistant to. The role of these factors in determining observed antibiotic resistance profiles suggests that any study measuring resistance in wild isolates should include the broadest possible range of bacterial species, host species and sampling locations. As such, this study provides an important new baseline for future measurements of antibiotic resistance in the Australian environment.

Aminoglycosides↗
PubMed2002

Surveillance of methicillin-resistant Staphylococcus aureus (MRSA) and Enterobacteriaceae producing extended-spectrum beta-lactamase (ESBLE) in Northern France: a five-year multicentre incidence study.

In order to measure the incidence of methicillin-resistant Staphylococcus aureus (MRSA) and of Enterobacteriaceae producing extended-spectrum beta-lactamase (ESBLE), and to evaluate the impact of the national guidelines for multidrug-resistant bacteria (MDRB) prevention in hospitals of Northern France, a multicentre study was conducted for three months every year starting in 1996, in volunteer hospital laboratories. All clinical specimens positive for MRSA and ESBLE were prospectively surveyed. During the five-year surveillance period, the overall proportion of MRSA was 38.4% in the 28,534 strains of S. aureus, and that of ESBLE was 11.4% in the 6121 strains of Klebsiella pneumoniae and 47.7% in the 2353 strains of Enterobacter aerogenes. The overall incidence rates of clinical specimens positive for MRSA, ESBL-K. pneumoniae and E. aerogenes were 0.84. 0.05 and 0.12/1000 hospital-days (HD), respectively. In the 23 hospitals that participated in the survey every year, the proportion and incidence of ESBLE decreased. Hence, despite recommendations as for isolation precautions, MRSA remains poorly controlled and requires more effective measures.

Anti-Bacterial Agents↗
PubMed1996

Allosteric regulation of the third ribonucleotide reductase (NrdEF enzyme) from enterobacteriaceae.

Enterobacteriaceae contain genes for three separate ribonucleotide reductases: nrdAB code for a class Ia enzyme, active during aerobiosis, nrdDG for a class III enzyme, active during anaerobiosis, and nrdEF for a cryptic class Ib enzyme. The NrdEF enzyme provides the active reductase in other, widely different bacteria. Here, we describe the allosteric regulation of the Salmonella typhimurium NrdEF enzyme. It consists of two tightly bound homodimeric proteins, R1E and R2F. Nucleoside triphosphates (ATP, dATP, dGTP, and dTTP) regulate the substrate specificity by binding to a single site of the R1E protein (one nucleotide per polypeptide). Regulation is similar to that of the NrdAB enzyme, with one major exception: dATP stimulates reduction of CDP (and UDP) under conditions when dATP strongly inhibits all activity of the NrdAB enzyme. The nrdA-coded R1 protein contains a second binding site for dATP (and ATP) that controls general enzyme activity. All known R1E proteins lack the 50 N-terminal amino acids of R1, and we propose that the activity site is located in this area of the protein. The more sophisticated regulation of NrdAB enzymes of eukaryotes provides protection against the possibly harmful overproduction of dNTPs.

Allosteric Regulation↗
PubMed2001

Diversity of polyamine patterns in soft rot pathogens and other plant-associated members of the Enterobacteriaceae.

Polyamine profiles of 91 pectolytic and other plant-associated strains from 30 taxa of the Enterobacteriaceae were obtained by gradient high performance liquid chromatography (HPLC). Pectobacterium carotovorum, basonym Erwinia carotovora, contained a high amount of putrescine and less diaminopropane. Diaminopropane was absent in Pectobacterium chrysanthemi, basonym E. chrysanthemi, whereas cadaverine was present in addition to the major compound putrescine. This chemotaxonomic difference reflects the deepest phylogenetic branching point within the recently emended genus Pectobacterium which lies between the two species P. carotovorum and P. chrysanthemi. Both important soft rot pathogens are easily distinguishable from each other and from the type species of the genus Erwinia as diaminopropane is the only major polyamine compound in E. amylovora. Chemotaxonomic heterogeneity is also emerging with respect to DYE's Amylovora group proposed in an early phytopathological concept.

Bacterial Typing Techniques↗
PubMed2003

Sequencing of 16S rDNA of Klebsiella: taxonomic relations within the genus and to other Enterobacteriaceae.

The 16S rDNAs of 20 strains of Klebsiella were sequenced and used for construction of a phylogenetic tree together with already published Enterobacteriaceae 16S rDNA sequences. The taxonomy within the Klebsiella genus, as reflected by the 16S rDNA tree, was in agreement with existing DNA-DNA hybridisation and numerical taxonomy data, indicating that for Klebsiella, 16S rDNA sequencing is a valid method for identification and taxonomical purposes. Five closely related clusters were found in the Klebsiella genus; Cluster I, K. oxytoca; Cluster II, K. terrigena, Cluster III, K. planticola and K. ornithinolytica; Cluster IV, Enterobacter aerogenes (K. mobilis); and Cluster V, K. pneumoniae. The position of Calymmatobacterium granulomatis within the genus and closest to K. pneumoniae was confirmed. For the species K. oxytoca, data seem to indicate a subdivision into two subspecies. In addition, a biochemically aberrant Klebsiella strain (BEC441) that was included in the analysis could not be assigned to any of the known species, but was found to be closest related to K. oxytoca. Furthermore, the high sequence similarity between the two environmental species K. planticola and K. ornithinolytica does not justify a distinction of the two species. Finally, within a 165-bp stretch of the 16S rDNA sequences, species-specific nucleotides were found.

DNA, Bacterial↗
PubMed1988

A preliminary investigation of antibiotic resistance in Enterobacteriaceae isolated from children with diarrhoea from four developing countries.

The prevalence of resistance to commonly used antibiotics was investigated in groups of children from four developing countries, Peru, Belize, Zaire and Sudan. Enterobacteriaceae spp. isolated from faeces of children with diarrhoea were tested for sensitivity to ampicillin, tetracycline, sulphonamide, trimethoprim, streptomycin and chloramphenicol. Overall, the highest prevalence of resistance was to sulphonamide (56% of children) and the lowest was to chloramphenicol (19% of children). For individual locations, isolates from Sudan had the highest prevalence of antibiotic resistance, 65% of the isolates being resistant to ampicillin, tetracycline, trimethoprim and streptomycin. Transfer of resistance was studied for some isolates using Escherichia coli Hb101 as recipient.

Belize↗
PubMed1993

Antibiotic resistance of Enterobacteriaceae isolated from the faecal flora of fattening pigs.

From June 1991 to April 1992 407 faecal samples were collected from three groups of pigs (I n = 248, II n = 87, III n = 72) at a pig fattening farm to determine the prevalence and the degree of antibiotic resistance of Enterobacteriaceae as well as the antibiotic susceptibility of the strains isolated. Despite the absence of mass medication during the observation period, the prevalence of resistance to the most commonly used antimicrobial agents in veterinary medicine was high (range amoxicillin 70%-97%, oxytetracycline 89%-100%, sulfamethoxazole 88%-100%, trimethoprim 78%-100%). The high degree of resistance to oxytetracycline and sulfamethoxazole ranged from 8%-67% and 4%-46%, respectively. The percentage of the isolated Escherichia coli strains resistant to oxytetracycline, streptomycin and sulfamethoxazole ranged from 49% to 68%; the other agents tested showed lower percentages (0-13%). Resistance to three or more antibiotics was observed in 43% of the isolates. Of the 52 resistance patterns that could be distinguished, 51% was accounted by only four patterns: oxytetracycline+streptomycin+sulfamethoxazole 20%, sulfamethoxazole 12%, streptomycin+sulfamethoxazole 11% and streptomycin+oxytetracycline 8%.

Animal Husbandry↗
PubMed2004

Risk factors for extended-spectrum beta-lactamase-producing Enterobacteriaceae in a neonatal intensive care unit.

Risk factors for colonization or infection with extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae during an outbreak in a neonatal intensive care unit (NICU) included low gestational age and exposure to third-generation cephalosporins. We also reviewed the existing medical literature regarding the clinical epidemiology of ESBLs in NICUs .

Analysis of Variance↗
PubMed1996

Diffusion of carbapenems through the outer membrane of enterobacteriaceae and correlation of their activities with their periplasmic concentrations.

Scarce information is available on the real mechanism by which carbapenemes penetrate in Enterobacteriaceae, although a considerable amount of evidence suggests that in many species of this family the lack of certain outer membrane proteins is associated with the acquisition of resistance to these antibiotics. The existance of specific pathways for the carbapenems has never been demonstrated, although at times it has been postulated in both wild and mutant strains, on the basis of evident discordances between permeability patterns and suceptibility data. By using the Zimmerman and Rosselet technique, which requires the strain under investigation to harbor a suitable beta-lactamase, the permeability of intact Escherichia coli and Enterobacter cloacae cells to meropenem and imipenem was investigated by transferring a constructed vector carrying the carbapenem hydrolyzing CphA metallo-beta-lactamase gene into the parental strains and their porin-deficient mutants. Reduced amounts of nonspecific porins significantly reduced the penetration of both carbapenems. The virtual absence of porins caused the MICs of meropenem to increase, mostly in Enterobacter cloacae, while it did not affected the MICs of imipenem. No evidence of specific porin pathways of the type described in Pseudomonas aeruginosa was found.

Bacterial Outer Membrane Proteins↗
PubMed1996

Emergence of resistance to beta-lactam agents in enterobacteriaceae species with group I beta-lactamases in Spain.

The contribution of induction and stable derepression of chromosomal group I beta-lactamases to beta-lactam antibiotics resistance was studied in clinical isolates of Enterobacteriaceae, collected from patients treated with these antibiotics. Multiple isolates of the same species from the same patient were characterized by different typing methods. Sonicated extracts of cells were assayed for chromosomal and plasmid-mediated beta-lactamases by isoelectric focusing and cloxacillin inhibition studies. The specific beta-lactamase activity, basal and induced with cefoxitin, was determined to differentiate strains with inducible or derepressed production of the enzyme. Induction of beta-lactamases was performed in each strain against the beta-lactams used in the therapy of each patient. Older penicillins resulted in a moderate to strong increase in beta-lactamase activity, whereas the results obtained with first-generation cephalosporins were species dependent. Expanded-spectrum cephalosporins were weak inducers of beta-lactamases. Indeed, the use of cefotaxime for treatment preceded the appearance of strains that produced chromosomal group I beta-lactamases constitutively. These strains showed a remarkable reduction in sensitivity to ureidopenicillins, carboxipenicillins, expanded-spectrum cephalosporins, and monobactams, but not to carbapenems.

Anti-Bacterial Agents↗
PubMed1981

Four-hour presumptive identification of Enterobacteriaceae from blood cultures.

A modification of the Minitek identification system for the four-hour identification of gram-negative bacilli recovered from blood cultures was developed. This method enabled our laboratory to identify correctly 99 of 100 consecutive isolates of all members of Enterobacteriaceae representing seven genera and nine species. All rapid identifications were confirmed using conventional biochemical methods. The procedure employs 11 Minitek biochemical discs inoculated with 0.1 ml of a dense organism suspension. Two-tenths milliliter of this suspension was added to a blank well for motility testing. In most instances, identification reactions were identical to those routinely used for the Minitek system. Only with the biochemical citrate disc does the interpretation vary from the Minitek standard. In addition to the blood culture isolates, 22 unknown organisms were also tested. All were correctly identified within four hours of the detection of visible turbidity in broth. This method offers a convenient, economic, and rapid means of identifying gram-negative bacilli isolated from blood cultures.

Bacteriological Techniques↗
PubMed1982

Comparison of two methods for same-day identification of Enterobacteriaceae.

Two commercial methods, API 20E (as modified for same-day enterobacterial identification) and Micro-ID, were evaluated for ability to provide useful same-day information of 368 clinically isolated Enterobacteriaceae. Organisms included Escherichia coli (54), Shigella (7), Edwardsiella tarda (1), Salmonella enteritidis (10), Citrobacter (30), Klebsiella (55), Enterobacter (68), Hafnia alvei (2), Serratia (33), Proteus (64), Morganella morganii (24), Providencia (18), and Yersinia enterocolitica (2). Methods were those of manufacturers without supplemental tests. API at five hours identified 78.5% of strains to species, 9.5% to genus only, 10.1% as part of a spectrum of identifications (SI), and 1.9% incorrect. Micro-ID at four hours yielded 90.0% correct identification to species and 3.3% to genus only, 4.0% SI, and 2.7% incorrect. API identification of many Serratia, Citrobacter, Providencia strains was to genus only; most incorrect results occurred in Serratia marcescens. Micro-ID identified most organisms to species; incorrect identifications were mainly S. marcescens and Klebsiella pneumoniae. Both systems provided excellent identification of E. coli. Both methods sacrifice a degree of accuracy that varies with the species tested, as compared to overnight systems, but both provide rapid information of potential clinical value.

Culture Media↗
PubMed1983

Comparison of automated and rapid manual methods for the same-day identification of Enterobacteriaceae.

The Vitek AMS automated instrument method for identification of Enterobacteriaceae was compared with two rapid manual methods intended for the same purpose, the Micro ID System and the API 20E Same-Day procedure, on a series of 400 consecutive fresh clinical isolates. Results were compared with identifications obtained using the API 20E System with overnight incubation and supplemental tube biochemicals (when needed). Both the final (8-hour) and a manually requested, presumptive 5-hour result from the AMS were compared with the 4-hour results provided by the Micro ID and the 5-hour results provided by the API. The Micro ID system proved to be the most rapid and accurate of the three test systems by correctly identifying 96.8% (387/400) of isolates. The API 20E using 5-hour readings identified 90.7% (363/400) of isolates, although 96.8% (387/400) could be identified if supplemental overnight tests were employed to separate profile codes with "good likelihood, but low selectivity." The AMS correctly identified 88.8% (355/400) isolates after 5 hours, and 95.0% (380/400) following 8 hours incubation.

Automation↗
PubMedDate not supplied

Effect of serum and blood on Enterobacteriaceae grown in the presence of subminimal inhibitory concentrations of ampicillin and mecillinam.

Bacteria from the strains of five species of Enterobacteriaceae grown in the presence of subminimal inhibitory concentrations of ampicillin or mecillinam formed into filamentous or round cells. These filamentous and round cells as well as normal control bacteria were incubated with either fresh human serum or blood, the bactericidal effects of which were then determined. In most cases, the bactericidal effect of either serum or blood on filamentous or round cells was less than the effect on control cells. In some cases, the effect on these drug-exposed cells was similar to that on control cells, but in no instance was the effect greater for the drug-exposed cells than for the control cells. However, in all cases in which the bactericidal effect of either serum or blood on the control cells was greater than or equal to 99%, the bactericidal effect on the drug-exposed cells was close to 90%. Although drug-exposed cells were not much more resistant to the bactericidal effect of serum or blood than were normal bacteria, they clearly were not more susceptible to these effects.

Amdinocillin↗
PubMed1973

The apparent conservation of the internal low efficiency promoter of the tryptophan operons of several species of Enterobacteriaceae.

The specific activities of the tryptophan biosynthetic enzymes were determined for eleven different species of Enterobacteriaceae grown under repressing and non-repressing conditions. In all the bacteria examined the multipliciy of derepression for the first two enzymes of the pathway was at least twofold greater than the multiplicity of derepression for the last three enzymes.

Enterobacteriaceae↗
PubMed1979

Effect of antibiotics on the adherence of enterobacteriaceae to human buccal cells.

Antibiotics at bacteriostatic concentrations (four times the minimal inhibitory concentration) decreased the attachment of Enterobacteriaceae to human buccal cells in vitro and augmented the reversibility of such adherence. These actions of antibiotics may influence colonization by nosocomial bacteria and the subsequent infection that they cause.

Anti-Bacterial Agents↗
PubMed1984

Incidence of trimethoprim-sulfamethoxazole-resistant enterobacteriaceae among transplant recipients.

Of 114 recipients of pancreatic, renal, and bone marrow transplants who were given trimethoprim-sulfamethoxazole (TMP-SMZ) for antimicrobial prophylaxis, 44 (39%) had a total of 52 fecal isolates of TMP-SMZ-resistant gram-negative bacilli. In most of these 44 patients, the resistant isolate was found at a concentration of greater than or equal to 10(6) organisms/ml of feces. Escherichia coli was the most frequent of the isolates, and Citrobacter freundii was the next most frequent. Eight of the 114 transplant recipients had gram-negative bacteremia; in six of these eight patients, a TMP-SMZ-resistant gram-negative bacillus was the etiologic agent of bacteremia. Four of the latter six patients had stool cultures analyzed prior to the detection of bacteremia; all four had high concentrations (greater than or equal to 10(8)/ml) of fecal TMP-SMZ-resistant E. coli one to 20 days before they were found to have E. coli bacteremia. In each of these instances, the E. coli isolates from the stool and the blood had similar antibiograms. These findings indicated that resistance to TMP-SMZ is becoming more prevalent and that the screening of patients for the presence of fecal TMP-SMZ-resistant Enterobacteriaceae prior to initiation of long-term therapy with this antimicrobial agent may be worthwhile.

Bone Marrow Transplantation↗
PubMed1983

Plasmid-determined beta-lactamases identified in a group of 204 ampicillin-resistant Enterobacteriaceae.

Information is presented on the plasmid-determined beta-lactamases identified in 204 strains of ampicillin-resistant Enterobacteriaceae. The type most frequently identified was TEM-1 (in 85.3% of the strains), followed by SHV-1 (14.70%). Two types of plasmid-determined beta-lactamase were identified in 20 strains; in 18 of them one of the two was TEM-1 and in 13, SHV-1 (the TEM-1 + SHV-1 combination was observed in 12 strains). In the 41 Klebsiella strains the most frequently identified enzyme was SHV-1 (in 28 of the strains) and the proportion of strains with two plasmid-determined beta-lactamases was higher than in the other species studied.

Ampicillin↗