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A rapid-kinetic study of the class C beta-lactamase of Enterobacter cloacae 908R.

The individual rate constants for acylation and deacylation (k2 and k3, respectively) of the class C beta-lactamase of Enterobacter cloacae 908R by ampicillin and carbenicillin have been determined. For several other beta-lactams, the value of k2 was too high to be determined and the k2/k3 ratio could be larger than 10,000. Branched pathways were also shown to occur with several penicillins and cephalosporins.

Enterobacter cloacae↗

Localization and physical mapping of a plasmid-borne 23-kb nif gene cluster from Enterobacter agglomerans showing homology to the entire nif gene cluster of Klebsiella pneumoniae M5a1.

A physical and genetical map of the plasmid pEA3 indigenous to Enterobacter agglomerans is presented. pEA3 is a 111-kb large plasmid containing a 23-kb large cluster of nif genes which shows extensive homology (Southern hybridization and heteroduplex analysis) to the entire nif gene cluster of Klebsiella pneumoniae (Kp) M5a1. All the nif genes on pEA3 are organized in the same manner as in K. pneumoniae, except nifJ, which is located on the left end of pEA3 nif gene cluster (near nifQB). A BamHI restriction map of pEA3 and a detailed restriction map of the 23-kb nif region on pEA3 is also presented. The nif genes of pEA3 showed a low level of acetylene reduction in Escherichia coli, demonstrating that these genes are functional and contain the whole genetic information required to fix nitrogen. The origin of vegetative replication (OriV) of pEA3 was localized about 5.5 kb from the right end of the nif gene cluster. In addition to pEA3, large plasmids from four other strains of E. agglomerans showed homology to all the Kp nif genes tested, indicating that in diazotrophic strains of E. agglomerans nif genes are usually located on plasmids. In contrast, in most of the free-living, nitrogen-fixing bacteria the nif genes are on chromosome.

DNA Restriction Enzymes↗

Influence of freezing-thawing and refrigeration on R-plasmid (pRPJ24) stability in Enterobacter cloacae 94R.

The effect of twenty freezing (-20 degrees C) and thawing cycles of Enterobacter cloacae 94R cells containing the R-plasmid pRPJ24 inoculated into broth and ground beef meat samples revealed no loss of resistance due to plasmid instability. In addition, low temperature storage at 4 degrees C did not produce any significant loss of the tetracycline and kanamycin resistances encoded on the pRPJ24 plasmid. The results of this study indicated that indigenous R-plasmids like pRPJ24 are stable in resident recipients like E. cloacae 94R in ground beef. However, the proportion of viable cells containing the pRPJ24 plasmid decreased significantly after 20 freezing-thawing cycles over 14 days incubation at 4 degrees C.

Animals↗

Biochemical characteristics of Enterobacter agglomerans and related strains found in buckwheat seeds.

Thirty strains of bacteria were randomly isolated and identified from buckwheat seeds. The phenotypic characteristics of these strains agree well with those of the Enterobacter agglomerans-Erwinia herbicola complex. On the basis of the difference in indole production and gas production from D-glucose, the isolates were divided into 3 phenotypic groups, viz. I, II and III. Twenty two strains were in phenotypic group 1, which is negative for indole production and gas production from D-glucose, and resembles Pantoea agglomerans. All six strains in phenotypic group II, which is positive for indole production and negative for gas production from D-glucose, were identified as Erwinia ananas. Two strains in phenotypic group III, which is negative for indole production and positive for gas production from D-glucose, were identified as Rahnella aquatilis.

Enterobacter↗

Serotypes and biochemical profiles of British hospital strains of Enterobacter cloacae in relation to site of infection and antibiotic susceptibility.

Comparisons were made between the O-serotype, API 20E profile, site of isolation and antimicrobial resistance of clinical isolates of Enterobacter cloacae. Correlations were found between autoagglutinable strains and urinary-tract infection, and API 20E profile 3305573 and strains isolated from blood. The proportion of strains sensitive to amikacin, gentamicin, cefotaxime, cefuroxime and trimethoprim were 100%, 93%, 91%, 83% and 89%, respectively. No individual resistances or patterns of resistance were associated with O-serotype or biochemical profile. Strains isolated from urinary-tract infections were the most resistant, 40% being resistant to five or more antimicrobials compared to 18%, 12% and 4% for strains from blood, wounds and sputum, respectively. There were no readily identifiable phenotypes within E. cloacae that possessed unique characteristics that could contribute to infections in hospitals.

Anti-Bacterial Agents↗

Outbreak of colonization of neonates with Enterobacter sakazakii.

An outbreak of colonization of 11 neonates with Enterobacter sakazakii occurred in a neonatal intensive care unit from the 10 September to 17 October 1984. During this period Ent. sakazakii was isolated from throat and rectal swabs and tracheal aspirates, but not from blood, of the neonates. The duration of colonization ranged from 2 to 8 weeks. The isolates were resistant to amikacin and to tobramycin, but sensitive to gentamicin. Four of the 11 colonized neonates had clinical signs of severe sepsis and one of meningitis and four died in spite of intensive chemotherapy. The source and the mode of spread of Ent. sakazakii remained unknown as it was not found on the hands of staff or in the inanimate environment of the unit. Ent. sakazakii may be implicated in severe infections in neonates and should be included when screening clinical specimens.

Bacteriological Techniques↗

Prior antimicrobial therapy and resistance of Enterobacter Citrobacter and Serratia to third generation cephalosporins.

During a 6-month period all inpatients from whom Enterobacter, Citrobacter or Serratia. had been isolated were reviewed and information on selected variables recorded. Two groups, one including 19 patients with organisms resistant to third generation cephalosporins and the other 111 patients with susceptible organisms were compared. In the initial analysis, the mean number of antimicrobials received in the prior 2 months was the variable most strongly associated with isolation of resistant organisms (2.6 +/- 1.5 vs 1.5 +/- 1.6; P = 0.002). When patients who had received no antimicrobials were omitted from the analysis, the mean number of antimicrobials was similar (2.6 +/- 1.5 vs 2.3 +/- 1.5; P = 0.19). Comparisons of antimicrobials received in the prior 2 months showed only cefoxitin (9/70 vs 7/19; P = 0.016) and cefotaxime (4/70 vs 5/19; P = 0.008) to be associated with isolation of resistant organisms. These data suggest that, at our institution, antimicrobial therapy with an extended spectrum cephalosporin is an important risk factor for subsequent acquisition of an organism resistant to third generation cephalosporins.

Cephalosporins↗

Enterobacter cloacae in a haematology/oncology ward--first impressions.

The observation that Enterobacter cloacae has emerged as a cause of infection in a haematology/oncology ward prompted a prospective study over an eight-month period. Sensitivity to antibiotics, plasmid screening and typing methods were used to describe the epidemiology of the organism which was isolated from 22 patients. There was evidence of a limited amount of cross infection. No common sources were found. A large number of distinct strains were identified. E. cloacae infection was associated with prolonged profound leucopenia and broad-spectrum antibiotic therapy.

Adolescent↗

Analysis of plasmid pattern in paediatric intensive care unit outbreaks of nosocomial infection due to Enterobacter cloacae.

Sequential outbreaks of nosocomial infection due to multiply-resistant Enterobacter cloacae occurred in September 1987, and between December 1988 and January 1989, in a paediatric intensive care unit. A total of eight neonates were affected and most had received ventilatory support. Initially, we were unable to determine whether the two outbreaks were caused by the same strain of E. cloacae. After applying plasmid profile analysis to identify epidemic strains, we established that the strain from the first outbreak was different from the second outbreak strain, as each had its own plasmid pattern. During the second outbreak, an environmental bacteriological survey was carried out. We found that the distilled water containers were contaminated with E. cloacae which had the same plasmid profile. After changing the distilled water containers and by reinforcement of aseptic techniques, the nosocomial outbreak was terminated.

Cross Infection↗

Rapid genotyping shows the absence of cross-contamination in Enterobacter cloacae nosocomial infections.

Restriction fragment length polymorphism analysis (RFLP) of total DNA and rDNA regions was used for the epidemiological evaluation of 10 Enterobacter cloacae nosocomial isolates obtained from nine patients in our hospital. Five of these patients were hospitalized during overlapping periods, thus raising the question of cross-contamination. A single biochemical pattern and antibiotic susceptibility profile was observed for all isolates but one. In contrast, based on the results of total DNA and rDNA RFLP patterns, the genetic unrelatedness of the isolates was clearly shown, thus excluding a common source of contamination or patient-to-patient transfer.

Adult↗

Application and assessment of cloacin typing of Enterobacter cloacae.

Three methods, O-serotyping, phage typing and susceptibility to bacteriocins, were used to type 357 clinical isolates of Enterobacter cloacae cultured from 219 patients. One hundred and sixty isolates were typed by serology and phage typing. When these two methods were used, primary classification of isolates was based on serology (65.7% typable) and phage typing for further subdivision (94.1% typable). When all the isolates were typed by cloacin susceptibility, 81.5% of them were typable. Maximum discrimination between cultures was achieved when the three methods were used together; no single method was sufficiently discriminatory. There was a close parallel between serotyping and bacteriocin lysis pattern. The latter was easy to perform and the results were achieved within 48 h. By applying this typing system two episodes of cross-infection were identified in a haematology/oncology unit and intensive care unit.

Bacteriophage Typing↗

Transducer related Enterobacter cloacae sepsis in post-operative cardiothoracic patients.

The use of intra-arterial pressure monitoring devices in patients undergoing major cardiac surgery is on the increase. Here we document an outbreak of Enterobacter cloacae septicaemia among seven post-operative cardiothoracic patients. Detailed investigations revealed the source of this nosocomial problem to be contaminated transducer heads. The need to follow strict aseptic measures in handling intra-arterial devices is emphasized, in order to minimize the morbidity and mortality in otherwise low-risk post-operative patients.

Cardiac Surgical Procedures↗

Molecular typing of Enterobacter cloacae by pulsed-field gel electrophoresis of genomic restriction fragments.

Eleven multiply resistant Enterobacter cloacae isolates were obtained from eight preterm neonates in a neonatal intensive care unit (NICU) of one hospital in Osnabrück, together with one sensitive strain from another infant. The presence of similar antibiograms and biotypes in 11 isolates prompted further characterization of the isolates by pulsed-field gel electrophoresis (PFGE) of NotI generated genomic restriction fragments. For assessment of the discriminatory power of this typing method 50 non-related strains were included in the study. Non-related strains demonstrated a marked variation in restriction fragment patterns and were clearly discriminated one from another. In contrast, 11 of 12 isolates from the NICU exhibited identical restriction profiles, indicating a cluster associated with nosocomial infection. Since the discriminatory power of PFGE is high, reproducibility is good, and all reagents and equipment for DNA fingerprinting are commercially available, this technique is proposed as a useful tool in the microbiology laboratory for investigating the epidemiology of E. cloacae strains.

Bacterial Typing Techniques↗

Postoperative endophthalmitis caused by an Enterobacter species.

Six patients (aged 8 to 75 years) who were operated upon during the same day developed bacterial endophthalmitis on the following day; seven eyes were affected. Two patients had intracapsular cataract extraction, one extracapsular lens extraction, two extracapsular cataract extraction with intraocular lens implantation and one repair of bilateral scleral or corneoscleral perforations. Vitreous cultures taken from six eyes were positive for an Enterobacter sp. Despite antibiotic treatment systemically, subconjunctivally and intravitreally, four eyes had to be eviscerated, while two eyes showed evidence of shrinkage (phthisis); only one eye retained useful vision (9/10). Cotton swabs used during surgery, prepared manually from cotton wool moistened with saline, were identified as the source of infection; the batch had not been subjected to the prescribed autoclaving process. The importance of proper sterile procedures in association with surgery is emphasized.

Adolescent↗

Enterobacter cloacae in a neonatal intensive care unit: account of an outbreak and its relationship to use of third generation cephalosporins.

After uneventful use of cefotaxime and ceftazidime as first line therapy for three years in our neonatal intensive care unit we isolated cephalosporin-resistant Enterobacter cloacae (CREC) strains which caused clusters of cases or colonization and/or serious neonatal infection. By using two or more typing methods, at least five different strains with similar patterns of antimicrobial sensitivities were identified. The results of a case-control study did not support the notion that the use of third generation cephalosporins was associated with colonization and infection by CREC. The outbreak was brought under control by interrupting the transmission of the epidemic strain D, by measures such as cohort nursing, diligent handwashing before and after procedures, and thorough environmental cleaning as well as by decontamination with glutaraldehyde after dismantling of the blood gas analyser believed to have acted as a persistent reservoir. Our experience highlights the danger of inadequate supervision and maintenance of equipment used for near-patient testing and the need to monitor such equipment not only in terms of its calibration and analytical performance but also microbiologically.

Blood Gas Analysis↗

Clinical and bacteriological survey after change in aminoglycoside treatment to control an epidemic of Enterobacter cloacae.

The effect of a change in the first line antibiotic treatment in a neonatal unit was studied. A total of 238 neonates (G1), admitted between 1 January and 31 July 1989, and treated with gentamicin, were compared with 398 (G2) admitted between 1 August 1989 and 31 July 1990 who received amikacin, in the combination of ampicillin plus an aminoglycoside. This change was implemented in an attempt to prevent the spread of an epidemic strain of Enterobacter cloacae resistant to third generation cephalosporins and all aminoglycosides, except amikacin. The change in treatment had no effect on the incidence of nosocomial infections [19.7% (G1) vs. 16.3% (G2) RR = 1.21 (0.86-1.70)], but the proportion of patients with nosocomial infections caused by the E. cloacae decreased (6.3% vs. 2.0% RR 3.14 CI 1.35-7.28). Certain trends in the bacterial ecology emerged: E. aerogenes and Enterococci increased in G2. The proportion of gentamicin-resistant strains such as E. cloacae or Staphylococci decreased and there was no increase in aminoglycoside-resistant strains, except in Escherichia coli, in which resistance to amikacin rose from 0 to 3%. This study illustrates the influence of antimicrobial therapy on the species and the resistance of strains isolated in nosocomial infections. It also highlights the need for epidemiologic surveillance, and poses the question of how best to modify antibiotic policy.

Amikacin↗

PCR based fingerprinting of Enterobacter cloacae.

An outbreak of lower respiratory tract infection with Enterobacter cloacae occurred in an intensive care unit in a university teaching hospital. Random amplification of polymorphic DNA (RAPD) was used to assist in the investigation of the outbreak. The technique was readily applied to this organism and permitted differentiation between strains which had identical biochemical profiles and antibiograms. The versatility of this technique makes it attractive for use in hospitals where fingerprinting of any one of the many Gram-negative rods associated with nosocomial infection may be required from time to time.

Cross Infection↗

Pyrolysis mass spectrometry of cephalosporin-resistant Enterobacter cloacae.

Thirteen clinical and four environmental isolates of third-generation cephalosporin-resistant Enterobacter cloacae (CREC) together with single isolates from the hands of a nurse and from a blood gas analyser were associated with two clusters of nosocomial infection. With an unrelated CREC isolate they had been typed by serotype, biotype, ribotype and phage-type and were examined by pyrolysis mass spectrometry (PYMS) as described here. PYMS data yielded two clusters, major and minor. All except one isolate in the major cluster corresponded to type group identity (serotype 07, biotype 62, ribotype D) which had caused neonatal sepsis and colonization. Multivariate analysis showed a homogeneous group consisting of this strain plus two outliers. The minor cluster included four different strains, one of which, serotype 03, biotype 62, ribotype C had caused excoriation of the buttocks and colonization.

Cephalosporin Resistance↗