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PubMed2001

Properties of an extracellular polysaccharide produced by a strain of enterobacter isolated from pond water.

A bacterium which was isolated from pond water and identified as Enterobacter cloacae produced a viscous extracellular polysaccharide when it was grown aerobically in a medium containing sucrose as a sole source of carbon. The maximum molecular weight of the polysaccharide was about 9.0 x 10(5). The polysaccharide was composed of fucose, galactose, glucose, and glucuronic acid in a molar ratio of 2:3:2:1, but the molecular weight and the molar ratio of the sugar component were different from those of the polysaccharide produced by the same species reported elsewhere.

Chromatography, Gas↗
PubMed2001

Isolation and characterization of the Enterobacter cloacae cheR mutant defective in phosphate taxis.

A chemotaxis-defective mutant of Enterobacter cloacae IFO3320, designated EC1, was isolated after N-methyl-N'-nitro-N-nitrosoguanidine (NTG) mutagenesis. Computer-assisted capillary assays showed that EC1 failed to show chemotactic responses to peptone and inorganic phosphate (Pi). Cloning and sequence analysis showed that EC1 is a cheR mutant, suggesting that Pi taxis by E. cloacae is dependent on a methyl-accepting chemotaxis protein(s) (MCP). EC1 was further mutagenized with NTG to construct cheR pstS and cheR pstA double mutants. A recombinant plasmid pECT01.2, which contained the E. cloacae cheR gene, restored the ability of these double mutants to show chemotaxis toward peptone but not Pi. These results suggest that the phosphate-specific transport (Pst) system, together with a MCP(s), is required for detecting Pi in E. cloacae.

Chemotaxis↗
PubMed2004

Functional difference between Sinorhizobium meliloti NifA and Enterobacter cloacae NifA.

The nifA gene is an important regulatory gene and its product, NifA protein, regulates the expression of many nif genes involved in the nitrogen fixation process. We introduced multiple copies of the constitutively expressed Sinorhizobium meliloti (Sm) or Enterobacter cloacae (Ec) nifA gene into both the nifA mutant strain SmY and the wild-type strain Sm1021. Root nodules produced by SmY containing a constitutively expressed Sm nifA gene were capable of fixing nitrogen, while nodules produced by SmY containing the Ec nifA gene remained unable to fix nitrogen, as is the case for SmY itself. However, transfer of an additional Sm nifA gene into Sm1021 improved the nitrogen-fixing efficiency of root nodules to a greater extent than that observed upon transfer of the Ec nifA gene into Sm1021. Comparative analysis of amino acid sequences between Sm NifA and Ec NifA showed that the N-terminal domain was the least similar, but this domain is indispensable for complementation of the Fix- phenotype of SmY by Sm NifA. We conclude that more than one domain is involved in determining functional differences between Sm NifA and Ec NifA.

Amino Acid Sequence↗
PubMedDate not supplied

Haemolymph and fat body metallo-protease associated with Enterobacter cloacae infection in the bloodsucking insect, Rhodnius prolixus.

Analysis of zymograms with SDS-polyacrilamide gel electrophoresis containing gelatin as substrate, and performed on samples of haemolymph or fat body taken from Rhodnius prolixus inoculated or not with Enterobacter cloacae, demonstrated distinct patterns of protease activities: (i) in the haemolymph two proteases were induced in insects inoculated with bacteria; (ii) two proteases were detected in the fat bodies derived from non-inoculated controls or insect inoculated with sterile culture medium; (iii) haemolymph and fat body had both the same apparent molecular weights proteases (46 and 56 kDa); and (iv) these enzymes were characterized as metallo-proteases. The association of these enzymes in Rhodnius infected with bacteria was discussed.

Animals↗
PubMed1999

Phenotyping and genotyping methods applied to investigate the relatedness of Brazilian isolates of Enterobacter cloacae.

In order to evaluate the resolving power of several typing methods to identify relatedness among Brazilian strains of Enterobacter cloacae, we selected twenty isolates from different patients on three wards of a University Hospital (Orthopedics, Nephrology, and Hematology). Traditional phenotyping methods applied to isolates included biotyping, antibiotic sensitivity, phage-typing, and O-serotyping. Plasmid profile analysis, ribotyping, and macrorestriction analysis by pulsed-field gel electrophoresis (PFGE) were used as genotyping methods. Sero- and phage-typing were not useful since the majority of isolates could not be subtyped by these methods. Biotyping, antibiogram and plasmid profile permitted us to classify the samples into different groups depending on the method used, and consequently were not reliable. Ribotyping and PFGE were significantly correlated with the clinical epidemiological analysis. PFGE did not type strains containing nonspecific DNase. Ribotyping was the most discriminative method for typing Brazilian isolates of E. cloacae.

Bacterial Typing Techniques↗
PubMedDate not supplied

[Resistance to cefepime in Enterobacter cloacae isolates from hospitals in Bogotá, Colombia].

OBJECTIVE: The aim of this research was to detect the presence of genes encoding beta-lactamases which can confer cefepime resistance on Enterobacter cloacae isolates, meaning that this antibiotic may be considered an important therapeutic alternative. MATERIALS AND METHODS: 28 E. cloacae isolates collected from 4 hospitals in Bogotá during 2003 were analysed. Extended spectrum beta-lactamases and cephalosporinase production were phenotypically determined. The presence of extended spectrum beta-lactamase genes was determined by PCR amplification. Mating assays were done to determine the possible transfer of bla-genes encoding cefotaximases. RESULTS: Microbiological tests detected 57% of extended-spectrum beta-lactamase-producing isolates. blaTEM, blasHv and blacTX-M genes were detected by PCR in 82% of the isolates. 7 out of 9 isolates carrying group 1 blacTX-M genes were in the resistant or intermediate range for cefepime. These isolates produced cefotaxime-resistant transconjugants. CONCLUSION: A relationship was found between resistance to cefepime and the presence of cefotaximases in E. cloacae. blacTx-M genes were present in 32% of the isolates, indicating an significant spread in the hospitals being studied. The facility of these genes' transfer between other species and Enterobacteria genera becomes an important reason for detecting them and controlling their spread in hospital settings. These results suggest a cautious use of cefepime for treating infection caused by E. cloacae strains which might produce CTX-M enzymes.

Anti-Bacterial Agents↗
PubMed2006

Quinoline derivatives as promising inhibitors of antibiotic efflux pump in multidrug resistant Enterobacter aerogenes isolates.

Efflux pumps protect the bacterial cell by expelling toxic compounds before they reach intracellular targets. Because this mechanism actively contributes to the resistance of a given bacterium to more than one class of antibiotics, molecules that are able to block the relevant efflux pump are of potential significance to combat drug resistance caused by efflux pumps. Different quinoline derivatives including alkoxy, alkylamino, thioalkoxy and chloroquinolines have been previously reported to make Enterobacter aerogenes resistant isolates that over express the mechanism of efflux, noticeably more susceptible to structurally unrelated antibiotics. In addition, various quinoline derivatives significantly increase the intracellular concentration of chloramphenicol as reported with other inhibitors, thereby suggesting the inhibition of the drug transport by AcrAB-TolC pump, which is fully active in the clinicaly resistant isolates investigated. Here, we discuss the respective properties of this molecular family, taking into account the recent insights into the structural data of AcrB pump.

ATP Binding Cassette Transporter, Subfamily B↗
PubMed1986

Enhancement of beta-lactamase production by glycine in Enterobacter cloacae ATCC.13047.

Beta-lactamase production in Enterobacter cloacae ATCC.13047 was enhanced by glycine, and the degree of enhancement was dependent on the concentration of glycine and the medium used. Maximum enhancement occurred with a concentration of 0.05 moles/l of glycine, and of 4 media examined enhancement was greatest on Isosensitest Agar. Enhanced beta-lactamase production evoked by glycine was compared with that following induction by various concentrations of cefoxitin. The amount of beta-lactamase produced under conditions of maximum glycine enhancement was higher than that produced by cultures fully induced by cefoxitin. Other differences in the characteristics of the enhancement of beta-lactamase production by glycine and beta-lactamase induced by cefoxitin suggested that the mechanisms of the 2 phenomena were different. The significance of this finding in terms of the interpretation of in vitro antimicrobial sensitivity tests with beta-lactam antibiotics is discussed.

Aerobiosis↗
PubMed1987

Enterobacter meningitis--treatment complicated by emergence of mutants resistant to cefotaxime.

A case of Enterobacter cloacae meningitis in a postoperative patient is reported. A slow response to cefotaxime necessitated the use of gentamicin and trimethoprim-sulfamethoxazole for cure. Two types of resistance in the strain of E. cloacae isolated to cefotaxime were demonstrated: an inducible beta-lactamase that likely was the cause of the poor response to cefotaxime and a constitutive beta-lactamase in a mutant strain detected by a disc susceptibility test.

Aged↗
PubMed1989

Enterobacter bacteremia: a review of 58 episodes.

This report reviews 58 episodes of enterobacter bacteremia in 55 patients during a 66-month period at a 750-bed community teaching hospital in USA. The median age of patients was 65 years and 38% of the patients were older than 70 years. All patients had underlying conditions: rapidly fatal (7%), ultimately fatal (33%) and non-fatal (60%). Nosocomial acquisition of bacteremia occurred in 72% of episodes. 20 episodes (34.5%) were polymicrobial bacteremia. The major portal of entry was the lungs (34%). The overall mortality was 69%. Factors that adversely influenced the mortality rate were nosocomial acquisition (p less than 0.02), rapidly fatal and ultimately fatal underlying conditions (p less than 0.02), and lungs as the portal of entry (p less than 0.05).

Adolescent↗
PubMed1989

Multiply beta-lactam resistant Enterobacter cloacae infections linked to the environmental flora in a unit for cardiothoracic and vascular surgery.

During the period March 1987-May 1988, postoperative infection or colonization with Enterobacter cloacae occurred in 9/379 (2.4%) patients who underwent cardiovascular surgery. Five of the patients were infected with multiply beta-lactam resistant E. cloacae, of whom 4 had been infected with an identical, resistant strain during intervals of months. This strain was also found in the environmental flora of the cardiovascular operating suite and in a sink reservoir in the surgery department. All 4 patients with the identical resistant strain had serious complications during the postoperative period with symptoms of septicaemia in 3, multiorgan failure and shock in 2, and mediastinitis in 3. The single resistant strain of a different serotype was also associated with severe postoperative complications. The 4 sensitive strains were all different serotypes. None caused septicaemia, one was associated with mediastinitis, another with an uncomplicated sternum infection, and 2 were from sputum. In the 3 latter patients with sensitive strains and few postoperative complications, cephalosporins had not been used during the pre- or postoperative period.

Adult↗
PubMed2003

NmcA carbapenem-hydrolyzing enzyme in Enterobacter cloacae in North America.

An imipenem-resistant Enterobacter cloacae isolate was recovered from the blood of a patient with a hematologic malignancy. Analytical isoelectric focusing, inhibitor studies, hydrolysis, induction assays, and molecular sequencing methods confirmed the presence of a NmcA carbapenem-hydrolyzing enzyme. This first report of NmcA detected in North America warrants further investigation into its distribution and clinical impact.

Adult↗
PubMed2006

Enterobacter cloacae outbreak and emergence of quinolone resistance gene in Dutch hospital.

An outbreak of Enterobacter cloacae infections with variable susceptibility to fluoroquinolones occurred in the University Medical Center Utrecht in the Netherlands in 2002. Our investigation showed that a qnrA1 gene was present in 78 (94%) of 83 outbreak isolates and that a qnrA1-encoding plasmid transferred to other strains of the same species and other species. The earliest isolate carrying this same plasmid was isolated in 1999. qnrA1 was located in a complex integron consisting of the intI1, aadB, qacEDelta1, sul1, orf513, qnrA1, ampR, qacEDelta1, and sul1 genes that were not described previously. On the same plasmid, 2 other class 1 integrons were present. One was a new integron associated with the bla(CTX-M-9) extended-spectrum beta-lactamase.

Anti-Bacterial Agents↗
PubMed2026

Genomic and functional characterization of novel therapeutic lytic bacteriophages targeting multidrug-resistant Enterobacter cloacae.

The alarming rates at which extensively drug-resistant (XDR) and pandrug-resistant (PDR) Enterobacter cloacae in hospitals are increasing has begun to severely limit treatment options, and thus the urgency for alternative interventions, including bacteriophage therapy. The purpose of the study was to isolate and molecularly characterize phages that can infect E. cloacae, and, furthermore, to assess the antimicrobial efficacy of the four novel lytic bacteriophages (MMRP1, MMRP2, MMRP3, and MMRP4) against antimicrobial-resistant E. cloacae isolates and to evaluate their potential as alternative therapeutic strategies. These novel phages were characterized by plaque morphology, transmission electron microscopy (TEM), host range testing, thermal and chloroform stability assays, bacterial reduction assays, and whole-genome sequencing (WGS). Among 27 clinical isolates, MDR, XDR, and PDR phenotypes were observed in 20 (74.1%), six (22.2%), and one (3.7%) isolates, respectively. All four phages produced clear lytic plaques (0.5-3.0 mm) with titers reaching up to 6 × 1010 PFU/mL, and the phage cocktail lysed 81.4% (22 of 27 isolates) of clinical isolates with high host specificity. TEM revealed that all four E. cloacae-infecting phages (MMRP1, MMRP2, MMRP3, and MMRP4) belong to the class Caudoviricetes, exhibiting icosahedral capsids, tailed morphology, and double-stranded DNA genomes, consistent with current ICTV classification criteria. Whole genome sequencing and comparative phylogenetic analysis further resolved the taxonomic placement of these phages at the family level, positioning MMRP1 within the family Demerecviridae and MMRP4 within the family Straboviridae. All phages were stable from -20 to 40 °C and were unaffected by exposure to chloroform. Phage cocktail reduced bacterial OD₆₀₀ to ≤ 0.3 within 4 h in the bacterial reduction test. WGS revealed large circular dsDNA genomes of ~132 kbp (MMRP1) and ~149 kbp (MMRP4), GC content of 38%, and modular architectures encoding structural, lytic, and replication gene modules. The most striking and highlighted suggestion that in vitro evaluation of MMRP1 and MMRP4 are highly recommended to more deeper future experimental studies to combat MDR E. cloacae nosocomial infections supported by genomic foundation and eventually, the possibility to be suitable for phage-engineering applications in clinical settings.

Enterobacter cloacae↗
PubMed1991

[Ecological behavior of plasmids and resistance to lead of Enterobacter agglomerans isolated from soil].

Soil samples taken monthly in 1990 from five parks outside Tokyo were examined for Enterobacter agglomerans. A total of 348 strains were isolated and 250 of them were tested for the presence of plasmids by DNA agarose gel electrophoresis. All the isolates carried at least two kinds of plasmids. Those isolated from July to September showed four or five kinds of plasmids (group I) and those isolated from January to June and from October to December showed two or three kinds of plasmids (group II). A majority of the plasmids detected were of 1,500 or fewer base pairs. The isolates were tested for the Pb2+ resistance; group I strains were more resistant to lead than group II strains. It is presumed that bacterial plasmids are related with the ecosystem of soil and the resistance to lead in E. agglomerans.

Drug Resistance, Microbial↗
PubMed2005

Identification of anrF gene, a homology of admM of andrimid biosynthetic gene cluster related to the antagonistic activity of Enterobacter cloacae B8.

AIM: To identify the gene (s) related to the antagonistic activity of Enterobacter cloacae B8 and to elucidate its antagonistic mechanism. METHODS: Transposon-mediated mutagenesis and tagging method and cassette PCR-based chromosomal walking method were adopted to isolate the mutant strain(s) of B8 that lost the antagonistic activity and to clone DNA fragments around Tn5 insertion site. Sequence compiling and open reading frame (ORF) finding were done with DNAStar program and homologous sequence and conserved domain searches were performed with BlastN or BlastP programs at www.ncbi.nlm.nih.gov. To verify the gene involved in the antagonistic activity, complementation of a full-length clone of the anrF gene to the mutant B8F strain was used. RESULTS: A 3 321 bp contig around the Tn5 insertion site was obtained and an ORF of 2 634 bp in length designated as anrF gene encoding for a 877 aa polyketide synthase-like protein was identified. It had a homology of 83% at the nucleotide level and 79% ID/87% SIM at the protein level, to the admM gene of Pantoea agglomerans andrimid biosynthetic gene cluster (AY192157). The Tn5 was inserted at 2 420 bp of the gene corresponding to the COG3319 (the thioesterase domain of type I polyketide synthase) coding region on B8F. The antagonistic activity against Xanthomonas oryzae pv. oryzae was resumed with complementation of the full-length anrF gene to the mutant B8F. CONCLUSION: The anrF gene obtained is related to the antagonistic activity of B8, and the antagonistic substances produced by B8 are andrimid and/or its analogs.

Base Sequence↗
PubMed2003

Enterobacter sakazakii: infectivity and enterotoxin production in vitro and in vivo.

Enterobacter sakazakii has been implicated as the causal organism in a severe form of neonatal meningitis, with reported mortality rates of 40 to 80%. Dried infant formula has been identified as a potential source of the organism in both outbreaks and sporadic cases. In this study, clinical and foodborne isolates of E. sakazakii were evaluated for enterotoxin production by the suckling mouse assay. In addition, suckling mice were challenged both orally and by intraperitoneal injection. Of 18 E. sakazakii strains evaluated, four were found to test positive for enterotoxin production. All strains of E. sakazakii were lethal to suckling mice at 10(8) CFU per mouse by intraperitoneal injection, while two strains caused death by the peroral route. In in vitro assays, CHO, Vero, and Y-1 cells demonstrated both cell lysis and rounding when exposed to E. sakazakii strain LA filtrates. This is the first report describing any putative virulence factors of E. sakazakii.

Administration, Oral↗
PubMed1980

Enterobacter cloacae outer membrane permeability to ceftizoxime (FK 749) and five other new cephalosporin derivatives.

The ability of ceftizoxime to penetrate the outer membrane was compared with those of five other new cephalosporins: cefotiam, cefuroxime, cefotaxime, cefmetazole and cefoxitin, using a clinical isolate of Enterobacter cloacae as a test strain. Estimation of permeability was performed by a method utilizing the inhibitory activities of the cephalosporins against beta-lactamase located in the periplasm. Of the cephalosporins tested, both ceftizoxime and cefmetazole gave remarkably high concentrations in the periplasm, several times higher than those of cefotaxime and cefoxitin and ten or more times higher than those of cefuroxime and cefotiam. The approximate permeability coefficient of ceftizoxime was also several times higher than those of cefotiam and cefmetazole and over ten times higher than those of cefoxitin, cefuroxime and cefotaxime.

Cefmetazole↗