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PubMed2003

The use of Enterobacter cloacae ATCC 43560 in the development of a two-phase partitioning bioreactor for the destruction of hexahydro-1,3,5-trinitro-1,3,5-s-triazine (RDX).

Research on the biodegradation of explosives has focussed exclusively on the treatment of contaminated soil and water. In the present work the anaerobic degradation of hexahydro-1,3,5-trinitro-1,3,5-s-triazine (RDX) by Enterobacter cloacae ATCC 43560 was investigated, and a two-phase partitioning bioreactor (TPPB) was developed for the destruction of pure, past-date munitions. TPPBs are characterized by a cell-containing aqueous phase, and an immiscible and biocompatible organic phase into which very large amounts of toxic and/or insoluble substrates can be dissolved. Based on equilibrium partitioning, the substrate is then transported to the cells, in response to their metabolic requirements, providing a means of demand-based substrate delivery, and high bioreactor productivity. Through consideration of the critical logP of E. cloacae, whether various classes of solvents could be used as sole carbon and energy sources, the capacity of various organics to dissolve RDX, and solvent cost, 2-undecanone was ultimately selected as the delivery solvent for the TPPB. Using this solvent, both batch and fed-batch operation of the TPPB were undertaken, and the volumetric degradation rate of RDX was found to be higher in this arrangement than any previous values reported in the literature. This work has demonstrated the potential of a method for the destruction of decommissioned munitions involving the dissolution of RDX in 2-undecanone, the use of the RDX-rich solvent as the second phase in a TPPB to degrade this explosive, and the subsequent recycling and re-use of the solvent.

Anaerobiosis↗
PubMed1987

Human isolates of Enterobacter intermedium.

The phenotypic characters of four clinical strains of Enterobacter intermedium (a species usually isolated from surface water and soil) are described and compared to those of E. cloacae and E. asburiae.

Child, Preschool↗
PubMed2003

Direct sequencing and PCR mapping of integrons reveals multiple class 1 integrons in the multiresistant strain Enterobacter cloacae SCH88040794.

We have characterized the variable region of three integrons found in the multiresistant strain Enterobacter cloacae SCH88040794, using polymerase chain reaction and direct sequencing of amplicons. The first integron has the gene cassette arrangement dfrAI-aadA1 in its variable region and the second aadB-oxa9. The third integron is In40 (aacA4-qacF-cmlB-oxa9), which has recently been described. The multiresistance expressed by this strain to amikacin, gentamicin, trimethoprim, sulfonamides, oxacillin, chloramphenicol and quaternary ammonium compounds is due, at least in part, to these three integrons.

Anti-Bacterial Agents↗
PubMed2003

Stable integration and expression of mosquito-larvicidal genes from Bacillus thuringiensis subsp. israelensis and Bacillus sphaericus into the chromosome of Enterobacter amnigenus: a potential breakthrough in mosquito biocontrol.

Previously, we have successfully integrated a spectinomycin/streptomycin resistance gene into Enterobacter amnigenus strain An11, a potential host for mosquito control, using in vivo recombination via homologous recombination (An11S4::Omega). We now report the successful transfer of two mosquito-larvicidal genes, cry4B from Bacillus thuringiensis subsp. israelensis and binary toxin genes from Bacillus sphaericus, into the host genome. To facilitate the screening procedure, the E. amnigenus derivative, An11S4::Omega, was used as a host. The integration of both toxin genes by two successive crossover events interrupted the Omega region yielding two integrants designated An11S4::cry4B and An11S4::Omega::bin, respectively. Differences in the integration efficiency of these toxin genes were observed. The presence of both genes in the target sites of the host genome was verified by PCR. Cry4B was expressed weakly from An11S4::cry4B, but no expression of the binary toxin gene could be detected from An11S4::Omega::bin. Nevertheless, these two integrants exhibited mosquito-larvicidal activity against Aedes and Culex, suggesting that both proteins were expressed, but at very low levels.

Animals↗
PubMed2003

Selenate reduction by Enterobacter cloacae SLD1a-1 is catalysed by a molybdenum-dependent membrane-bound enzyme that is distinct from the membrane-bound nitrate reductase.

Enterobacter cloacae SLD1a-1 is capable of reducing selenium oxyanions to elemental selenium under both aerobic and anaerobic conditions. In this study the enzyme that catalyses the initial reduction of selenate (SeO4(2-)) to selenite (SeO3(2-)) has been localised to isolated cytoplasmic membrane fractions. Experiments with intact cells have shown that the putative selenate reductase can accept electrons more readily from membrane-impermeable methyl viologen than membrane-permeable benzyl viologen, suggesting that the location of the catalytic site is towards the periplasmic side of the cytoplasmic membrane. Enzyme activity was enhanced by growing cells in the presence of 1 mM sodium molybdate and significantly reduced in cells grown in the presence of 1 mM sodium tungstate. Non-denaturing polyacrylamide gel electrophoresis (PAGE) gels stained for selenate and nitrate reductase activity have revealed that two distinct membrane-bound enzymes catalyse the reduction of selenate and nitrate. The role of this membrane-bound molybdenum-dependent reductase in relation to selenate detoxification and energy conservation is discussed.

Benzyl Viologen↗
PubMed2004

Diversity of chromosomal AmpC beta-lactamases from Enterobacter cloacae isolates in a Portuguese hospital.

Six clinical isolates of Enterobacter cloacae isolated in a Portuguese hospital, between April 1999 and November 2000, demonstrated resistance to almost all broad-spectrum cephalosporins, except to cefepime. These isolates were susceptible to quinolones and to aminoglycosides. Isoelectric focusing demonstrated production of beta-lactamases with pIs > 8.0 and by all six isolates, exhibiting a cephalosporinase phenotype. The results of pulsed field gel electrophoresis revealed that these isolates were genetically unrelated. The amino acid sequence of six AmpC beta-lactamases (Eclo1FF, Eclo6FF, Eclo9FF, Eclo10FF, Eclo11FF and Eclo15FF) shared 97-99% homology with the chromosomal AmpC beta-lactamase from E. cloacae P99 and 86-87% homology with those of two plasmid-mediated AmpC beta-lactamases, MIR-1 and ACT-1. This is the first report of chromosomal AmpC beta-lactamase production by E. cloacae isolates in a Portuguese hospital.

Amino Acid Sequence↗
PubMed2005

[Enterobacter cloacae spondylodiscitis through misuse of high-dose intravenous buprenorphine].

INTRODUCTION: We report a case of Enterobacter cloacae spondylodiscitis related to risk practices in intravenous drug addicts (IVDA). OBSERVATION: The patient, a former heroin addict, was receiving long-term, high-dose buprenorphine maintenance treatment. He had been misusing the treatment, injecting it daily for several months. The clinical course included several uncommon features that are usually found in IVDA patients: subacute infection, apyrexia, and minimal inflammatory syndrome. This infection also led to the discovery of his HIV infection. DISCUSSION: Any dorsolumbar pain in IVDA patients, including those receiving regular drug maintenance treatment and especially those with HIV infection, should suggest spondylodiscitis, because of these patients' enhanced sensitivity to infection and the frequent bacteremia caused by persistent or transitory relapse involving injection (exchange of material, reuse of needles, syringes, cotton swabs, and risk of contamination through the hands or saliva).

Adult↗
PubMedDate not supplied

Separation of Escherichia adecarboxylata from the "Erwinia herbicola-Enterobacter agglomerans" complex and from the other Enterobacteriaceae by nucleic acid and protein electrophoretic techniques.

The species Escherichia adecarboxylata was examined for DNA relatedness to the "Erwinia herbicola-Enterobacter agglomerans" complex and to other members of the family Enterobacteriaceae. DNA-DNA hybridizations (nitrocellulose filter method) showed that strains received as E. adecarboxylata were highly related to each other (73-100% homology). Three strains of E. agglomerans and one strain of E. herbicola showed, respectively, 77, 96, 97 and 92% relatedness with the labelled DNA of E. adecarboxylata. Two groups (E2 and E3) of "atypical coliforms" previously described by Gavini et al. (1983) showed high reassociation values (76-79% and 80-89%, respectively) with E. adecarboxylata. Most of these strains produced similar or nearly identical protein electrophoregrams. All these strains were therefore classified in E. adecarboxylata. This taxon yielded hybridization values lower than 53% with the previously described phenetic or genetic groups belonging to or related to the "herbicola-agglomerans" complex and values lower than 64% with 56 other species of the Enterobacteriaceae. It was concluded that E. adecarboxylata is a species different from E. agglomerans and the other species of the family Enterobacteriaceae. A new definition of the species E. adecarboxylata is presented.

Bacterial Proteins↗
PubMedDate not supplied

Genetic engineering of Enterobacter aerogenes with the Vitreoscilla hemoglobin gene: cell growth, survival, and antioxidant enzyme status under oxidative stress.

Hemoglobins in unicellular organisms, like the one here in the bacterium Vitreoscilla, have greater chemical reactivity than their homologues in multicellular organisms. They can catalyze redox reactions and may protect cells against oxidative stress. The ability of Vitreoscilla hemoglobin to complement deficiencies of terminal cytochrome oxidases in Escherichia coli also suggests that this hemoglobin can receive electrons during respiration. In this study, a recombinant strain of Enterobacter aerogenes engineered to produce the Vitreoscilla Hb was investigated with regard to its susceptibility to oxidative stress. The culture response to oxidative stress produced by exogenously applied hydrogen peroxide was characterized in terms of cell growth, survival and the activities of two key antioxidant enzymes (catalase and superoxide dismutase). The influence of the physiological state of the cells and different media upon these culture dynamics was determined. Results showed that the hemoglobin-expressing strain is quite distinct in terms of growth/survival properties and activity of antioxidant enzymes from that of non-hemoglobin counterparts.

Bacterial Proteins↗
PubMedDate not supplied

Relationship between the physiology of Enterobacter agglomerans CNCM 1210 grown anaerobically on glycerol and the culture conditions.

In a preliminary study, levels of activity of enzymes involved in anaerobic glycerol catabolism by Enterobacter agglomerans grown in batch cultures regulated in a pH range of 6.5-8.0 were monitored. That study showed that activities of key enzymes of the downstream metabolism of glycerol--glyceradehyde-3-phosphate dehydrogenase (GAP-DH), lactate dehydrogenase and pyruvate formate lyase--were strongly dependent on the culture pH. To investigate the influence of pH on the physiology of the strain, E. agglomerans was grown anaerobically in a continuous culture supplied with glycerol as the sole carbon source and regulated at pH 8. A complete biochemical analysis was performed and was compared with that previously described for the continuous culture regulated at pH 7. A limitation of the glycolytic flux at the level of GAP-DH was demonstrated at high dilution rate, resulting in an overflow metabolism through the 1,3-propanediol formation pathway. Increasing the specific rate of glycerol consumption also resulted in enhanced lactate production due to limitation by the pyruvate decarboxylation step. Finally, changing the culture pH significantly modified the enzymatic profile of E. agglomerans, and it enabled the stability of the culture to be increased by preventing the accumulation in the fermentation broth of 3-hydroxypropionaldehyde, an inhibitory metabolite, when the glycerol supply was suddenly increased.

Acetyltransferases↗
PubMed2000

Spectrum and transferability of beta-lactam resistance in hospital strains of Enterobacter isolated in Bratislava and Innsbruck.

The transferability and expression of beta-lactam resistance were compared in multiresistant clinical isolates of Enterobacter spp. collected from different hospitals in Bratislava, Slovakia (n = 15) and Innsbruck, Austria (n = 19) during 1996-1997. The strains from Bratislava were resistant to ampicillin, cefoxitin, cefotaxime, ceftazidime and ceftriaxone. All strains from Innsbruck were resistant to ampicillin and cefoxitin; 17 were also resistant to ceftazidime and aztreonam but the majority remained susceptible to cefotaxime and ceftriaxone. All strains were susceptible to cefepime and imipenem. The majority of the tested strains transferred resistance determinants to E. coli recipient by conjugation. Production of beta-lactamase including ESBL was the major mechanism of beta-lactam resistance. Large plasmids of 77-88 and 91 kb were confirmed in clinical isolates from Bratislava and Innsbruck.

Anti-Bacterial Agents↗
PubMed2002

Growth of cell-wall-deficient variants of Enterobacter cloacae facilitates beta-lactamase derepressed mutants.

The degree to which cell-wall-deficient bacteria (CWDB) are involved in the generation of beta-lactamase derepressed mutants (DM) was measured using Enterobacter cloacae 3624. The frequency of DM in non-permissive isotonic ticarcillin medium was compared with their frequency in hypertonic ticarcillin medium that supports CWDB growth. DM were resistant to extended spectrum penicillins and cephalosporins and had a basal beta-lactamase activity of >300 units/mg protein. Anaerobic growth of CWDB increased the relative risk of DM 2 x 10(6)-fold. Aerobic incubation produced fewer CWDB colonies but the risk of DM was still increased 400-fold over non-permissive controls. These results define a new role for CWDB as intermediaries in the emergence of resistance.

Anti-Bacterial Agents↗
PubMed2003

Thanatin activity on multidrug resistant clinical isolates of Enterobacter aerogenes and Klebsiella pneumoniae.

Efflux pumps protect bacterial cells by ejecting intracellular toxic molecules such as antibiotics, detergents and defensins that have penetrated the cell envelope. Some of these efflux pumps recognise structurally unrelated compounds (mdr pump) and account for the resistance of some organisms to two or more agents. It would be of interest to identify molecules that are able to circumvent the problems created by multidrug resistance phenotypes during antibiotic therapy. We have studied the activity of thanatin, a 21-residue cationic antimicrobial peptide produced by an insect, against three bacterial species. The antibacterial effect depended on the size of lipopolysaccharide side chains. In clinically resistant isolates of Enterobacter aerogenes and Klebsiella pneumoniae, the biological activity of thanatin is independent of the membrane permeability, possibly controlled by one or more porins, and/or the expression of drug efflux pumps, two mechanisms which confer high level antibiotic resistance. In addition, thanatin was able to improve the activity of structurally unrelated antibiotics (norfloxacin, chloramphenicol, tetracycline) on a multidrug- resistant E. aerogenes clinical isolate.

Anti-Bacterial Agents↗
PubMed2003

4-alkoxy and 4-thioalkoxyquinoline derivatives as chemosensitizers for the chloramphenicol-resistant clinical Enterobacter aerogenes 27 strain.

Enterobacter aerogenes is a Gram-negative bacteria frequently responsible for nosocomial respiratory tract infections. Strains resistant to chloramphenicol are frequently isolated. Alkoxy and thio-alkoxyquinolines have a potential to act as chemosensitizers that would render multi-drug-resistant (MDR) bacterial infections susceptible to antibiotics to which they were originally resistant. Several new quinoline derivatives have been prepared, characterized and studied for their ability to increase chloramphenicol sensitivity of E. aerogenes 27, a clinical strain that exhibits the MDR phenotype. Drugs investigated were either quinoline ethers or quinoline thio-ethers. Thio-ethers are much more efficient in increasing chloramphenicol sensitivity than other corresponding ethers. In particular, 4-piperidinoethylthio-quinoline increases the strain sensitivity to chloramphenicol by about 20 times at 2 mM concentration. Similarly, sensitivity to quinolone antibiotics dramatically increases. Because these quinoline derivatives act as inhibitors of the drug efflux pump responsible for bacterial resistance to chloramphenicol, they may serve as adjunct to conventional therapy of E. aerogenes infections.

Anti-Bacterial Agents↗
PubMed1997

ERIC-PCR typing profiles of Enterobacter cloacae are stable after development of advanced cephalosporin resistance.

Enterobacterial repetitive intergenic consensus (ERIC) sequence polymerase chain reaction genotype profiles of Enterobacter cloacae were prepared for paired isolates which were obtained before and after the high level expression of the chromosomal cephalosporinase. These profiles were homologous within strain pairs which suggests that the acquired resistance does not lead to major chromosomal alterations. ERIC genotypes are therefore reliable markers in epidemiological investigations where antibiogram modifications may arise as a consequence of antibiotic exposure and beta-lactamase over-production..

Bacterial Typing Techniques↗
PubMed1999

The effect of new disinfectant substances on the metabolism of Enterobacter cloacae.

The antimicrobial mechanism of 16 new commercially manufactured disinfectant substances on an Enterobacter cloacae strain was studied. The substances tested represent 11 quaternary ammonium salts (QAS) and five combinated QAS with other ingredients. The antimicrobial efficacy was characterized by influencing the growth of bacterial cells expressed by MIC and ED50 values as well as by the inhibition of the incorporation rate of [14C] adenine and [14C] leucine. The disinfectants are divided into three groups according to their efficacy. The first group comprised substances with strong inhibitory effect (MIC 0.006-0.048 mg l(-1)) such as triquart, topax 91, benzalkonium chloride, neoquat S, ID 213, and antibacteric P. The second group represented substances with good antibacterial efficacy (MIC 0.048-0.15 mg l(-1)), and the third group were substances with MIC values up to 0.195-0.39 mg l(-1). Cetrimide had low activity (MIC 3.12-6.25 mg l(-1)). The effect of substances studied on the biosynthetic processes expressed by R values (IC50 Ade:IC50 Leu) showed that these values were < 1 except ADL 007. Much lower IC50 Ade and IC50 Leu values of the disinfectant substances studied suggested interference of these substances with nucleic acid synthesis and proteins synthesis which was expressed by inhibition of both precursors. All substances except cetrimide caused an inhibition of the endogenous respiration. The highest inhibition was caused by benzalkonium chloride. This affected the respiration significantly in the presence of intermediators of the Krebs' cycle (glycerol, aspartate). The tested substance suppressed the growth of E. cloacae probably through interference with energy-yielding and energy-requiring processes of the cells.

Carbon Radioisotopes↗
PubMed1996

The role of N-actylglucosaminyl-1,6 anhydro N-acetylmuramyl-L-alanyl-D-glutamyl-meso-diaminopimelic acid-D-alanine for the induction of beta-lactamase in Enterobacter cloacae.

The mechanism of beta-lactamase induction in Enterobacter cloacae which is linked to the peptidoglycan recycling, was investigated by HPLC analysis of cell wall fragments in genetically defined cells. It is demonstrated here that the transmembrane protein AmpG transports not only the precursor muropeptide of M-tripeptide (N-acetylmuramyl-L-alanyl-D-glutamyl-meso-diaminopimelic acid), the D-tripeptide (N-actylglucosaminyl-1,6 anhydro N-acetylmuramyl-L-alanyl-D-glutamyl-meso-diaminopimelic acid), but also that of M-tetra-peptide (N-acetylmuramyl-L-alanyl-D-glutamyl-meso-diaminopimelic acid-D-alanine), the D-tetrapeptide (N-actylglucosaminyl-1,6 anhydro N-acetylmuramyl-L-alanyl-D-glutamyl-meso-diaminopimelic acid-D-alanine), into the cytoplasm. These findings indicate that probably also M-tetrapeptide and D-tetrapeptide are signal muropeptides for beta-lactamase induction. In fact, D-tetrapeptide, not D-tripeptide, increases upon imipenem treatment.

Acetylmuramyl-Alanyl-Isoglutamine↗
PubMed1999

Survival of Enterobacter cloacae and Pseudomonas paucimobilis in yoghurts manufactured from cow's milk and soymilk during storage at different temperatures.

The survival of two microbial contaminants, Enterobacter cloacae and Pseudomonas paucimobilis, in yoghurts manufactured from cow's milk and soymilk was investigated during storage for 45 days at 4 and 12 degrees C. Sensory panel tests performed before microbiological investigation, showed that the flavor of soy-yoghurts made with cocoa powder or malt added did not have the beany taste of soy beans. Both contaminants were significantly resistant to low pH values during storage for 32 days at 4 degrees C. The survival at 4 degrees C was remarkable in both plain and flavored yoghurts and a population close to 10(2) C.F.U./ml was observed after 38 days of storage. Experiments performed with soymilk yoghurts showed an enhanced survival of P. paucimobilis at 4 degrees C compared to the storage in cow's milk yoghurts; microbial values were close to 7-8 x 10(6) C.F.U./ml after 16 days. Soymilk exhibited a protective effect on L. delbrueckii subsp. bulgaricus and S. thermophilus at 12 degrees C and, compared to the survival in cow's milk yoghurts, a larger number of viable cells of both probiotic microorganisms (10(6) and 10(8) C.F.U./ml, respectively) were observed after 36 days of storage.

Animals↗