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PubMed2001

[Enterobacter cloacae epidemic on a neonatal intensive care unit due to the use of contaminated thermometers].

From December 1999 to March 2000 a nosocomial outbreak of multiresistant Enterobacter cloacae occurred in the neonatal intensive care unit (NICU) at the VU Medical Center, Amsterdam, the Netherlands. Twenty-six patients were infected or colonized with this strain resistant to third generation cephalosporins and with decreased sensitivity for aminoglycosides. Three neonates experienced sepsis with E. cloacae with serious clinical symptoms and two of them died. Comparison of the Enterobacter isolates by amplified-fragment length polymorphism indicated that this outbreak was caused by the spread of a single strain. Infection control precautions were initiated in order to stop further spread; barrier precautions, enforcement of hand disinfection and cohorting of colonized patients. A multidisciplinary crisis team coordinated these infection control precautions and informed all persons involved. Analysis of antibiotic usage in 1999 showed an increase in the use of third generation cephalosporins from November onwards. Due to the resistance pattern of the epidemic strain the use of third generation cephalosporins was discontinued in February 2000. At the end of February the NICU was temporarily closed. The epidemic strain of E. cloacae was isolated from one digital rectal thermometer. Patient use of thermometers and disposable coverings for rectal thermometers were introduced to eliminate this possible means of spread. No spread of multiresistant E. cloacae was found following the introduction of these interventions. Once all the neonates had been transferred, the NICU was disinfected and reopened in March.

Cephalosporin Resistance↗
PubMedDate not supplied

[Beta-lactam resistance in aquatic Enterobacter cloacae strains using phenotypic and genotypic criteria].

Bacterial resistance by producing of beta-lactamases represents an increasing problem of infections chemotherapy. beta-lactam hydrolyzing activities are detected in virtually all bacteria, from witch Enterobacter cloacae produce chromosomal beta-lactamases included in inducible class AmpC beta-lactamases. The purpose of this preliminary study was to investigate the antibiotic susceptibility of 7 inducible beta-lactamase-producing Enterobacter cloacae strains isolated from aquatic sources (river and polluted waters). The identification to the species level was performed with the API 32E system and susceptibility to antimicrobial agents was tested by the disk diffusion method according to NCCLS recommendations. The following antibiotics were tested: ampicillin, amoxycillin/clavulanic acid, ceftazidime, cefotaxime, cephalotin, cefamandole, cephaclor, imipenem, amikacin, gentamycin, kanamycin, tobramycin, ciprofloxacin, norfloxacin, ofloxacin, nalidixic acid, tetracycline and chloramphenicol. Minimum inhibitory concentrations (MICs) were performed using dilution method in Mueller-Hinton broth with a 0.06-64 micrograms/ml concentration range for all antimicrobials and bacterial inoculum of about 1-2 x 10(8) cfu/ml. For the data analysis NCCLS breakpoints for resistance and sensitivity were used. Interaction of beta-lactamase inhibitor clavulanate with cefotaxime was performed by double-disk synergy test. Detection of inducible beta-lactamase expression was performed by the inductibility disk diffusion test using cefotaxime, ceftazidime and imipenem. Genomic DNA was isolated using CTAB technique and bacterial plasmid isolation was performed by an alkaline lysis method. Genetic characterization was performed by agarose gel electrophoresis and spectrophotometric analysis. The majority of examined E. cloacae strains were sensitive to imipenem, cefamandole, amikacin and quinolones (norfloxacin and ofloxacin), a higher moderate resistance being observed only to nalidixic acid (higher than 50%) and ciprofloxacin (15%). The percentage of resistant strains ranged from 72% (to kanamycin) to 87% (to gentamicin). The best results (resistance about 99%) were obtained with ampicillin, amoxycillin/clavulanic acid, ceftazidime, cefotaxime, cephalotin, cephaclor, tobramycin, tetracycline and chloramphenicol. The disk diffusion tests showed the absence of extended-spectrum beta-lactamases production and the expression of inducible beta-lactamases. Electrophoretic patterns point out the presence of plasmid DNA. Plasmid profile revealed the presence of several different plasmids ranging from 2.5 kpb to more than 30 kpb. The presence of inducible beta-lactamase E. cloacae strains in aquatic media (river and polluted waters) and the closely related pattern of susceptibility among these strains reflect the possible contamination of these sources and the common origin of them.

Anti-Bacterial Agents↗
PubMed1994

[Ribosomal DNA fingerprinting analysis of Enterobacter cloacae isolated from an outbreak of nosocomial infection].

16s and 23s ribosomal DNA (rDNA) genes were amplified form E. coli, labelled with [alpha- 32P] dATP by nick translation, and applied as conserved gene probes to detecting broad-spectrum rDNA. rDNA fingerprinting analysis of Enterobacter cloacae (E. cloacae) isolated form an outbreak of nosocomial infection and other hospital environmental sources was conducted with digestion by different restrictive endonuclease, such as Hind III, EcoRI, BamHI, Bgl I, et al. Results showed rDNA fingerprinting of E. cloacae strains isolated from 10 patients with nosocomial infection was the same as that from other sources. It suggested E. cloacae causing this outbreak was originated from one genetic clone, and its source of infection was the tubes and humidifying bottles for oxygenation. rDNA fingerprinting technique is accurate, reliable, specific and repeatable, and will plays important roles in identification and classification of bacteria, tracking the source of transmission of nosocomial infection.

Cross Infection↗
PubMed1995

[An investigation into antibiotic resistance and plasmid pattern of Enterobacter cloacae in burn infection].

In view of a high positive isolation rate and resistance to antibiotics of Enterobacter cloacae in our department, 32 strains of E. cloacae, which were isolated from blood and infected wounds of burned patients from June through November 1990, were randomly selected to determine their susceptibility to 21 antibiotics and their plasmid patterns. It was found that 29 of 32 strains were resistant to at least nine antibiotics, while the other three strains were relatively sensitive. 27 of 32 strains of E. cloacae were tested for plasmid, and among them, three strains (relatively sensitive to antibiotics) contained no plasmid. The plasmid patterns of the other 24 isolates varied with strains, but all of them had a 80 kb plasmid. The 80 kb plasmid was obviously related to the high isolation rate and high resistance to antibiotics of E. cloacae isolated in our department.

Burns↗
PubMed1998

Prenatal urinary ascites and persistent cloaca: risk factors for poor drainage of urine or meconium.

PURPOSE: The unique confluence of the urinary, genital and gastrointestinal tracts in girls with persistent cloaca potentially results in aberrant drainage of urine and meconium that may be characteristic, allowing a prenatal diagnosis to be made. MATERIALS AND METHODS: Three of our last 15 patients with cloacal anomalies presented with urinary ascites due to intraperitoneal reflux through the genital system. In 3 female patients a long, narrow cloaca was associated with a high confluence of the urethra, vagina and rectum. All 3 patients ultimately underwent creation of a cutaneous vesicostomy. RESULTS: In all 3 cases cutaneous vesicostomy effectively diverted urine away from the cloaca before definitive repair. CONCLUSIONS: The urethra-like persistent cloaca may result in significant outflow resistance. When combined with high confluence, it may cause aberrant drainage of urine and/or meconium. When such problems are noted prenatally, they are likely to persist postnatally and require intervention.

Ascites↗
PubMed1990

A method for removal of toxic chromium using dialysis-sac cultures of a chromate-reducing strain of Enterobacter cloacae.

A method for removal of toxic hexavalent chromium (chromate: CrO2-4) was developed by use of dialysis-sac cultures of a chromate-reducing strain of Enterobacter cloacae (HO1). E. cloacae strain HO1 cells were put in dialysis (semipermeable membrane) sacs, and the sacs were submerged in water containing toxic CrO2-4. The dialysis sacs allowed CrO2-4 to diffuse into the culture, and CrO2-4 was reduced anaerobically in the dialysis sacs by the E. cloacae cells. Because reduced chromium readily formed insoluble chromium hydroxides in the dialysis sacs, the greater part of reduced chromium was unable to diffuse out through the semipermeable membrane. Thus the dialysis culture of E. cloacae strain HO1 could successfully remove toxic chromium from the surrounding water. If the initial concentration of CrO2-4 was less than 4 mM (208 ppm as chromium), about 90% of the total chromium could be removed from water by the described method.

Bacteriological Techniques↗
PubMed1985

Plasmid-determined cloacin DF13-susceptibility in Enterobacter cloacae and Klebsiella edwardsii; identification of the cloacin DF13/aerobactin outer membrane receptor proteins.

Both Enterobacter cloacae H478 and Klebsiella edwardsii S15 were shown to harbour a relatively large conjugative plasmid that coded for cloacin DF13-susceptibility and the production and uptake of a hydroxamate iron chelator, most probably aerobactin. Protein-blotting experiments with antiserum raised against the purified cloacin DF13/aerobactin receptor protein from Escherichia coli (Co1V-K30) revealed that the corresponding outer membrane receptor proteins of Ent. cloacae H478 and K. edwardsii S15 had apparent mol wts of 85 000 and 76 000, respectively. E. coli transconjugants harbouring either the plasmid from Ent. cloacae H478 or K. edwardsii S15 expressed a cloacin DF13/aerobactin outer membrane receptor protein with a mol wt of 74 000. The receptor protein encoded by the Ent. cloacae and K. edwardsii plasmids were immunologically more related to each other than to the pCo1V-K30-encoded receptor protein.

Bacterial Outer Membrane Proteins↗
PubMed1990

Chromate-resistance in a chromate-reducing strain of Enterobacter cloacae.

Resistance to toxic hexavalent chromium (chromate: CrO4(2)) in Enterobacter cloacae strain HO1, isolated from an activated sludge sample, was investigated under aerobic and anaerobic conditions. Decreased uptake of 51CrO4(2-) in E. cloacae strain HO1 was observed under aerobic conditions, when compared with a standard laboratory E. cloacae strain (IAM 1624). Under anaerobic conditions E. cloacae strain HO1 was able to reduce hexavalent chromium to the less toxic trivalent form. When E. clocacae strain HO1 was grown with nitrate anaerobically, the cells were observed to lose simultaneously their chromate-reducing ability and chromate-resistance under anaerobic conditions.

Anaerobiosis↗
PubMedDate not supplied

Biochemical characterization of type A and type B beta-lactamase from Enterobacter cloacae.

Different types of chromosomally coded beta-lactamases are found in Enterobacter cloacae. E. cloacae M6300 produces beta-lactamase type A, which has an isoelectric point of 8.8, whereas E. cloacae 908 R produces beta-lactamase type B, which has an isoelectric point of 7.9. Both enzymes were purified to homogeneity by a procedure that included affinity chromatography on amino phenylboronic acid-modified Sepharose. The two enzymes were closely related as shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, kinetic constants with several substrates, amino acid composition, NH2-terminal amino acid sequence, and reaction with antisera. In addition to having different isoelectric points, the two enzymes migrated to slightly different positions on polyacrylamide gels and differed significantly in rate of catalysis for cephalothin, imipenem, and the penem Sch 34343. One of three antisera seemed to recognize an epitope that differs in the two enzymes. The diversity of cephalosporinases found in E. cloacae with respect to the evolution of novel beta-lactamases was considered.

Amino Acid Sequence↗
PubMedDate not supplied

Impact of the ampD gene and its product on beta-lactamase production in Enterobacter cloacae.

In an investigation of the influence of the ampD gene on beta-lactamase production and induction in Enterobacter cloacae, the ampR-ampC gene region cloned into a plasmid and the ampD gene cloned into another vector were transferred to a strain of Escherichia coli. The genetically manipulated E. coli strains served as a model for study of the inducibility of beta-lactamases in E. cloacae. In addition, beta-lactamase induction in E. cloacae bearing the previously mentioned plasmids was studied. After induction of the beta-lactamase with cefoxitin, the specific hydrolytic activity, the viable cell count, and the degradation of cefoxitin were determined. beta-Lactamase expression decreased with an increasing amount of the ampD gene product. The cefoxitin concentration decreased in proportion to the amount of enzyme, but the induction of beta-lactamase seemed not to be an important factor influencing the viable cell count of E. cloacae as long as cefoxitin concentrations exceeded the MIC. Despite different beta-lactamase concentrations, the decrease in the viable cell count was nearly identical in all experiments.

Cefoxitin↗
PubMed1987

Isolation and selection of a bacteriophage-typing set for Enterobacter cloacae.

Seventy-six phages active against Enterobacter cloacae were isolated from sewage and other sources. They were tested at RTD on 92 selected strains of E. cloacae and their lytic reactions were used to select phages for a typing set. Numerical analysis by the Jaccard coefficient was used to assess the similarity between the phages. A computer-based test selection procedure selected sub-sets of phages to discriminate between all 92 E. cloacae strains and within the most frequent serological groups. A subjective analysis of the candidate phages based on similarity, clarity of plaque and frequency of lysis was combined with the computer selected sub-sets to produce a final set of 25 phages that gave a good theoretical discrimination of clinical isolates of E. cloacae.

Bacteriophage Typing↗
PubMed1982

Outer membrane permeation of beta-lactam antibiotics in Escherichia coli, Proteus mirabilis, and Enterobacter cloacae.

Mutant strains lacking outer membrane protein(s) were isolated from Escherichia coli, Proteus mirabilis, and Enterobacter cloacae. The outer membrane protein(s) of P. mirabilis and E. cloacae corresponding to E. coli porin were identified on the basis of their function, namely, their ability to allow the permeation of glucose as demonstrated by [14C]glucose uptake by intact cells. P. mirabilis has only one outer membrane pore protein (molecular weight, 40,000), but E. cloacae has at least two such proteins (molecular weights, 37,000 and 39,000 to 40,000). When the bacteria lost these proteins or porin, the outer membrane permeation of cefazolin was found to be greatly reduced in these three species. Such a change in the outer membrane permeation closely correlated with a significant decrease in the bacterial susceptibility to cephalosporins, including cefoxitin. These results suggested that the main pathway for cephalosporin permeation is the pore made up of these proteins. The 39,000- to 40,000-molecular-weight protein in E. cloacae was also assumed to play an important role in the outer membrane permeation of tetracycline and chloramphenicol. On the other hand, the permeation route of penicillins was obscure. The susceptibility to penicillins, except in some cases, was little influenced by the absence of the proteins. Ampicillin was found to pass through the outer membrane via the same route as the cephalosporins, but the possibility that ampicillin and other penicillins possess another unknown route for outer membrane permeation could not be ruled out.

Anti-Bacterial Agents↗
PubMed1991

Altered phenotypes associated with ampD mutations in Enterobacter cloacae.

A study was done to determine the genetic locus responsible for altered expression of AmpC beta-lactamase in Enterobacter cloacae 1194E and several mutants derived from E. cloacae 029. These phenotypes were defined by units of enzyme activity found in sonic extracts of cells before and after induction with cefoxitin and included (units uninduced/units induced) the wild-type (7/219), high-level constitutive (10,911/10,862), temperature-sensitive (at 30 degrees C 82/706 and at 42 degrees C 5,031/6,020), and hyperinducible (19/1,688) phenotypes. When the ampD region of each E. cloacae strain was cloned and introduced into an ampD mutant Escherichia coli strain, the altered phenotypes were found to reside within this locus. Furthermore, transformants containing wild-type ampD were poorly inducible at 42 degrees C while those with high-level constitutive or hyperinducible ampD were unaffected by temperature. Since the source of ampD was the only variable in these E. coli transformants, these results suggested that ampD encodes a protein that is involved in sensing the inducer. To test this possibility, the responses to different inducers of E. coli transformants containing various ampD regions were assessed. In the presence of wild-type ampD, transformants responded equally to cefoxitin and cefotetan, regardless of temperature. In the presence of temperature-sensitive ampD, induction by cefotetan was similar to that by cefoxitin at 30 degrees C but greater than that by cefoxitin at 42 degrees C. These results suggest that ampD encodes a protein involved in induction of AmpC beta-lactamase in E. cloacae.

Anti-Bacterial Agents↗
PubMed1989

Enumeration of Enterobacter cloacae after chloramine exposure.

Growth of Enterobacter cloacae on various media was compared after disinfection. This was done to examine the effects of monochloramine and chlorine on the enumeration of coliforms. The media used were TLY (nonselective; 5.5% tryptic soy broth, 0.3% yeast extract, 1.0% lactose, and 1.5% Bacto-Agar), m-T7 (selective; developed to recover injured coliforms), m-Endo (selective; contains sodium sulfite), TLYS (TLY with sodium sulfite), and m-T7S (m-T7 with sodium sulfite). Sodium sulfite in any medium improved the recovery of chloramine-treated E. cloacae. However, sodium sulfite in TLYS and m-T7S did not significantly improve the detection of chlorine-treated E. cloacae, and m-Endo was the least effective medium for recovering chlorinated bacteria. Differences in recovery of chlorine- and chloramine-treated E. cloacae are consistent with mechanistic differences between the disinfectants.

Chloramines↗
PubMed1993

Cloning and Characterization of Aerobactin Biosynthesis Genes of the Biological Control Agent Enterobacter cloacae.

Five strains of Enterobacter cloacae that are biological control agents of Pythium damping-off diseases produced the hydroxamate siderophore aerobactin under iron-limiting conditions. Genes determining aerobactin biosynthesis of the biocontrol strain E. cloacae EcCT-501 were localized to a 12.3-kb region, which conferred aerobactin production to Escherichia coli DH5alpha. The aerobactin biosynthesis genes of E. cloacae hybridized to those of the pColV-K30 plasmid of E. coli, but restriction patterns of the aerobactin regions of pColV-K30 and E. cloacae differed. A derivative strain with a deletion in the aerobactin biosynthesis locus was as effective as strain EcCT-501 in biological control of Pythium damping-off of cucumber. Thus, aerobactin production did not contribute significantly to the biological control activity of EcCT-501 under the conditions of this study.

Journal Article↗
PubMed1984

Transfer of the chromosomal bla gene from Enterobacter cloacae to Escherichia coli by RP4::mini-Mu.

The resistance gene for beta-lactamase-stable cephalosporins from Enterobacter cloacae was transferred to Escherichia coli by the aid of RP4::mini-Mu. The R-prime plasmids generated carried 60 to 80 kilobases (kb) of E. cloacae DNA and coded for the chromosomal E. cloacae beta-lactamase. The gene was fully expressed in the recipient. Restriction endonuclease EcoRI fragments of the R-prime plasmid pBP100 were cloned into the vector pBP328, yielding the plasmid pBP102 with a size of 14 kb. A restriction map of this plasmid was constructed. By digesting pBP102 into seven PstI fragments, ligating the fragments, and looking for the smallest plasmid generated, pBP103 was isolated. It consisted of three PstI fragments, two of them (together 4.2 kb) necessary for resistance. During the experiment (performed in a recA+ background) the largest PstI fragment had undergone a substitution of a 0.3-kb segment of pBP102 by a 0.7-kb segment in pBP103 (as deduced by heteroduplex analysis). The bla gene of resistant E. cloacae strains was dominant over the gene of susceptible organisms.

Cephalosporins↗
PubMed1987

Common mechanism of ampC beta-lactamase induction in enterobacteria: regulation of the cloned Enterobacter cloacae P99 beta-lactamase gene.

Expression of the chromosomal beta-lactamase from the ampC gene in inducible in both Enterobacter cloacae and Citrobacter freundii. Cloning of ampC as well as its regulatory gene, ampR, from E. cloacae P99 revealed a gene organization indentical to that of C. freundii in the corresponding region. Although almost no similarities could be found between the restriction maps of ampC and ampR in the two species, the genes cross-hybridize. Also, both ampR gene products have a size of about 31,000. The regulatory features of E. cloacae beta-lactamase induction are very similar to those in C. freundii, i.e., beta-lactamase synthesis is repressed by AmpR in the absence, and stimulated in the presence, of inducer. The AmpR function can be transcomplemented between the two species, but there are quantitative regulatory aberrations in such hybrids, in contrast to the total complementation obtained within each system. These results suggest that the mechanism of beta-lactamase induction is the same in E. cloacae, C. freundii, and other gram-negative bacteria with inducible chromosomal beta-lactamase expression.

Cloning, Molecular↗
PubMed1968

Purification and genetic determination of bacteriocin production in Enterobacter cloacae.

Enterobacter cloacae (strain DF13) was found to produce a bacteriocin which could be induced by mitomycin C. In the supernatant fluid of the induced culture phagelike particles were found. The bacteriocin was partially purified from induced cultures by ammonium sulfate precipitation and gel-filtration on Sephadex G-150. Ultraviolet-absorbing material was eluted from the Sephadex column in three fractions. The biological activity was mainly present in the second fraction and is associated with a protein with a molecular weight of about 61,000. The phagelike particles were found in the first fraction and show no biological activity. Upon conjugation of E. cloacae strain DF13 with another strain of the same species and with Escherichia coli K-12S, the ability to produce bacteriocin was transferred. The new bacteriocinogenic strain produced bacteriocin, which could not be distinguished from that produced by E. cloacae strain DF13. Although transfer of the bacteriocinogenic factor often occurred together with transfer of the ability to produce phagelike particles, it was shown that these two factors are two separate genetic entities. In addition to a bacteriocinogenic factor, E. cloacae strain DF13 was found to carry two other transferable plasmids: one determining resistance against streptomycin and sulfanilamide and another determining resistance against penicillin.

Bacterial Proteins↗