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PubMed1984

Characterization of two clinical, multiple-drug-resistant isolates of Enterobacter cloacae.

Two multiple drug resistant Enterobacter cloacae isolates (Nos. 460 and 493) varied phenotypically in bacteriocin susceptibility in the absence of significant O antigen variation. Both isolates were susceptible to chloramphenicol, nitrofurantoin, polymyxin B, nalidixic acid, norfloxacin, and enoxacin only. One isolate carried a non-conjugative resistance (R) plasmid, whereas the other isolate contained a conjugative, 'curable' R plasmid and a cryptic plasmid. Both wild-type isolates constitutively produced a chromosomal cephalosporinase (nitrocefin hydrolysis); 'cured' variants of E. cloacae isolate No. 493, which had become susceptible for lamoxactam, produced a cefazolin-inducible beta-lactamase. The two E. cloacae isolates, including their 'cured' variants, were of low-grade virulence for outbred NMRI mice. Both isolates differed somewhat in susceptibility to defibrinated human blood. Inhibitory (0.25 microgram/ml), but not subinhibitory (0.125 microgram/ml) concentrations of norfloxacin and enoxacin combined with human blood yielded additive effects against both E. cloacae isolates.

Adult↗
PubMed1989

[Nosocomial infections with resistant Enterobacter cloacae. Effects on the disease and the treatment].

We describe five patients with serious nosocomial infection caused by resistant Enterobacter cloacae. Four of the patients underwent cardiovascular operations and were infected with an identical, multiple beta-lactam resistant E cloacae strain. The fifth patient was admitted with several wounds and skin-ulcers containing both resistant and sensitive E cloacae of other types. The resistant strain caused septicemia in four patients; three developed multiorgan failure and one died. The most probable causes of these infections were an abundant and constant reservoir of E cloacae in a sink at the operating unit, periodical breakdown of barriere routines, and heavy use of cephalosporins.

Adult↗
PubMed2004

Location of the ureteral openings in the cloacas of tinamous, some ratite birds, and crocodilians: a primitive character.

Cloacas of 67 avian species, of both sexes, from various habitats and differing dietary habits, were examined macro- and microscopically to investigate possible variation in the location of the ureteral openings. Differing from most birds studied, in adult male Rhea americana and several tinamous species the ureters were found to open into the coprodeum. In these species the urodeum receives only the vas deferens or oviduct. Similarly, in crocodiles Caiman crocodilus yacare, but not in lizards Tropidurus montanus and snakes Crotalus durissus terrificus, the ureters empty into the coprodeum. This similarity between ancient birds (ratites and tinamous) and crocodiles may indicate a primitive character linking reptiles and birds. This unusual position of the ureteral orifice can represent an adaptation to facilitate urine collection into the coprodeum and large intestine. Another possibility is that this variation in ureter position is a male reproductive strategy to avoid the mixture of urine and semen in the cloaca. There were no evident correlations between the location of the ureteral openings and the birds' habitat, diet, or histology of the coprodeal mucosa. The occurrence of a phallus in eight species of birds was detected, as well as a peculiar vascularization related to the coprodeal epithelium of anseriformes. Together, these data add to the scarce information about the morphophysiology of the avian cloaca, and also contribute to clarify avian phylogenetic linkages.

Animals↗
PubMed2001

Quantitative structure-activity relationships in two-electron reduction of nitroaromatic compounds by Enterobacter cloacae NAD(P)H:nitroreductase.

Enterobacter cloacae NAD(P)H:nitroreductase (NR; EC 1.6.99.7) catalyzes the reduction of a series of nitroaromatic compounds with steady-state bimolecular rate constants (kcat/Km) ranging from 10(4) to 10(7) M(-1) s(-1). In agreement with a previously proposed scheme of two-step four-electron reduction of nitroaromatics by NR (Koder, R. L., and Miller, A.-F. (1998) Biochim. Biophys. Acta 1387, 395-405), 2 mol NADH per mole mononitrocompound were oxidized. An oxidation of excess NADH by polinitrobenzenes, including explosives 2,4,6-trinitrotoluene (TNT) and 2,4,6-trinitrophenyl-N-methylnitramine (tetryl), has been observed as a slower secondary process, accompanied by O2 consumption. This type of "redox cycling" was not related to reactions of nitroaromatic anion-radicals, but was caused by the autoxidation of relatively stable reaction products. The initial reduction of tetryl and other polinitrophenyl-N-nitramines by E. cloacae NR was analogous to a two-step four-electron reduction mechanism of TNT and other nitroaromatics. The logs kcat/Km of all the compounds examined exhibited parabolic dependence on their enthalpies of single-electron or two-electron (hydride) reduction, obtained by quantum mechanical calculations. This type of quantitative structure-activity relationship shows that the reactivity of nitroaromatics towards E. cloacae nitroreductase depends mainly on their hydride accepting properties, but not on their particular structure, and does not exclude the possibility of multistep hydride transfer.

Aniline Compounds↗
PubMed1994

Development of resistance by Enterobacter cloacae during therapy of pulmonary infections in intensive care patients.

The emergence of resistance during therapy and the efficacy of different antibiotic therapy regimens were studied in 38 intensive care patients suffering from pulmonary infections caused by Enterobacter cloacae. Every three days a fresh isolate was obtained from each patient and tested in vitro for susceptibility to 16 antibiotics by determination of the minimal inhibitory concentrations. During therapy with cefotaxime and tobramycin the E. cloacae strains from 47% of the patients became resistant to cefotaxime within 6 days. In all cases resistance encompassed all other broad-spectrum penicillins and cephalosporins tested, as well as aztreonam. Development of resistance regularly led to persistence of bacteria. Resistance to tobramycin, ciprofloxacin or imipenem was not observed. Treatment of 25 patients with persisting E. cloacae infections was successful in 17 out of 18 patients treated with imipenem and in 6 out of 7 patients receiving ciprofloxacin.

Adolescent↗
PubMed2005

Enterobacter cloacae, an obligatory endophyte of pollen grains of Mediterranean pines.

Enterobacter cloacae was found to be associated with the pollen of several Mediterranean pines. The bacterium was detected only in mature pollen of Pinus halepensis, P. brutia, and P. pinea. E. cloacae is considered to be an obligatory endophyte based on its occurrence in disinfected male cones and the successful inoculation of seedlings of the above 3 species with E. cloacae AS1 isolated from pollen of P. halepensis used as a model strain. Strain AS1 was able to produce indolyl-3-acetic acid (IAA) from L-tryptophan in culture, and this was probably the source of the increased IAA content in the germination medium of pollen. In addition, strain AS1 promoted adventitious root formation in mung bean (Vigna radiata) cuttings. However, it was not possible to obtain bacterium-free pollen to elucidate its role in pollen germination.

Colony Count, Microbial↗
PubMed2006

Purification, characterization, and gene cloning of 4-hydroxybenzoate decarboxylase of Enterobacter cloacae P240.

We found the occurrence of 4-hydroxybenzoate decarboxylase in Enterobacter cloacae P240, isolated from soils under anaerobic conditions, and purified the enzyme to homogeneity. The purified enzyme was a homohexamer of identical 60 kDa subunits. The purified decarboxylase catalyzed the nonoxidative decarboxylation of 4-hydroxybenzoate without requiring any cofactors. Its Km value for 4-hydroxybenzoate was 596 microM. The enzyme also catalyzed decarboxylation of 3,4-dihydroxybenzoate, for which the Km value was 6.80 mM. In the presence of 3 M KHCO3 and 20 mM phenol, the decarboxylase catalyzed the reverse carboxylation reaction of phenol to form 4-hydroxybenzoate with a molar conversion yield of 19%. The Km value for phenol was calculated to be 14.8 mM. The gene encoding the 4-hydroxybenzoate decarboxylase was isolated from E. cloacae P240. Nucleotide sequencing of recombinant plasmids revealed that the 4-hydroxybenzoate decarboxylase gene codes for a 475-amino-acid protein. The amino acid sequence of the enzyme is similar to those of 4-hydroxybenzoate decarboxylase of Clostridium hydroxybenzoicum (53% identity), VdcC protein (vanillate decarboxylase) of Streptomyces sp. strain D7 (72%) and 3-octaprenyl-4-hydroxybenzoate decarboxylase of Escherichia coli (28%). The hypothetical proteins, showing 96-97% identities to the primary structure of E. cloacae P240 4-hydroxybenzoate decarboxylase, were found in several bacterial strains.

Amino Acid Sequence↗
PubMed2006

Mutation in cyaA in Enterobacter cloacae decreases cucumber root colonization.

Strains of Enterobacter cloacae show promise as biological control agents for Pythium ultimum-induced damping-off on cucumber and other crops. Enterobacter cloacae M59 is a mini-Tn5 Km transposon mutant of strain 501R3. Populations of M59 were significantly lower on cucumber roots and decreased much more rapidly than those of strain 501R3 with increasing distance from the soil line. Strain M59 was decreased or deficient in growth and chemotaxis on most individual compounds detected in cucumber root exudate and on a synthetic cucumber root exudate medium. Strain M59 was also slightly less acid resistant than strain 501R3. Molecular characterization of strain M59 demonstrated that mini-Tn5 Km was inserted in cyaA, which encodes adenylate cyclase. Adenylate cyclase catalyzes the formation of cAMP and cAMP levels in cell lysates from strain M59 were approximately 2% those of strain 501R3. Addition of exogenous, nonphysiological concentrations of cAMP to strain M59 restored growth (1 mM) and chemotaxis (5 mM) on synthetic cucumber root exudate and increased cucumber seedling colonization (5 mM) by this strain without serving as a source of reduced carbon, nitrogen, or phosphorous. These results demonstrate a role for cyaA in colonization of cucumber roots by Enterobacter cloacae.

Adenylate Cyclase Toxin↗
PubMed2006

Occurrence of yeasts in cloacae of migratory birds.

Several species of yeast have been reported as pathogens in humans based on increases in immunodeficiency syndromes and as a result of immunosuppressant chemotherapy in cancer treatment. Domestic and wild birds are known to act as carriers of human pathogenic fungi. To gain additional information on the yeasts present in the cloacae of some species of migratory birds, 421 wild birds (24.39% out of 1726 birds caught in Romania, Hungary and Bulgaria) were sampled with the permission of the local judicial authority. The state of conservation of the birds (i.e. post-mortem alterations, colour of the mucosae etc.), along with their age and sex were determined. Samples were collected directly from the cloacae and cultured, and colonies were identified in each positive sample. Yeasts were isolated from 15.7% of the animals sampled, with the highest percentage found in coots (Fulica atra -58.8%) and the lowest in quails (Coturnix coturnix -1.7%). A total of 131 isolates belonging to 15 species of yeast were identified. Rhodotorula rubra was the yeast with the highest number of isolates (28.2%), followed by Cryptococcus albidus (18.4%), Candida albicans (9.2%), Trichosporon cutaneum (8.4%), Candida guilliermondii (6.1%), Candida tropicalis (6.1%) and other species. The present study represents the first survey on the occurrence of yeasts in the cloacae of migratory birds. The prevalence and species of yeasts isolated is discussed on the basis of the ecology, diet, and habitat of the birds.

Animals↗
PubMed1991

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↗
PubMed1991

Hygienic hand disinfection for the removal of epidemic vancomycin-resistant Enterococcus faecium and gentamicin-resistant Enterobacter cloacae.

An outbreak strain of Enterococcus faecium, bearing plasmid-mediated vancomycin resistance, and an epidemic gentamicin-resistant, multiply-resistant strain of Enterobacter cloacae, both survived well on fingertips of three volunteers for up to 30 minutes after inoculation. Hand disinfection of inoculated fingers with 60% isopropyl alcohol, or with alcoholic chlorhexidine, reliably gave a 4 log10 reduction of both test organisms. Enterobacter cloacae could not be detected in any finger washings, even by enrichment culture. Chlorhexidine digluconate and povidone-iodine were also effective against E. faecium and Ent. cloacae, giving 4 log10 reductions, but finger washings taken after disinfection yielded low counts of the test strains, e.g. less than 45 recoverable colony forming units (cfu) per finger. Handwashing with soap and water was the least reliable method. The epidemicity, serious antimicrobial multiple resistance and survival on finger-tips of these two strains justifies the use of handwashing agents which have maximum effect. The rapid bactericidal (and residual) activity of alcoholic chlorhexidine suggests that, on the basis of present evidence, it is the preferred agent for hygienic hand disinfection against such strains.

Adult↗
PubMed1994

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↗
PubMed2005

Prevalence of decreased susceptibility to carbapenems among Serratia marcescens, Enterobacter cloacae, and Citrobacter freundii and investigation of carbapenemases.

Between March and July 2002, total of 612 clinical isolates of Serratia marcescens, Enterobacter cloacae, and Citrobacter freundii (201 S. marcescens, 228 E. cloacae, and 183 C. freundii) were collected from 13 clinical laboratories in a nationwide distribution. Imipenem and meropenem minimum inhibitory concentrations (MICs) were determined using the agar dilution method according to the National Committee for Clinical Laboratory Standards guidelines. For the isolates with a decreased susceptibility to carbapenems (MICs of >or=2 microg/mL), isoelectric focusing, polymerase chain reaction (PCR) amplification of the carbapenemase genes (bla(IMP-1), bla(VIM-2), bla(SME-1), bla(OXA-23), bla(OXA-25), bla(KPC-1)), and sequencing were performed. The prevalence of S. marcescens, E. cloacae, and C. freundii with a decreased susceptibility to imipenem was 17.9% (36/201), 0.4% (1/228), and 0.5% (1/183), respectively, and to meropenem, it was 11.4% (23/201), 0% (0/228), and 0.5% (1/183), respectively. The bla(VIM-2) was the only carbapenemase detected, and was found in 0.5% (1/201) of S. marcescens and 0.5% (1/183) of C. freundii isolate.

Anti-Bacterial Agents↗
PubMed2006

Ceftobiprole medocaril (BAL5788) treatment of experimental Haemophilus influenzae, Enterobacter cloacae, and Klebsiella pneumoniae murine pneumonia.

Ceftobiprole (BAL9141) is an investigational cephalosporin active against methicillin- and vancomycin-resistant staphylococci administered as a water-soluble prodrug, ceftobiprole medocaril (BAL5788). Using an immunocompetent murine pneumonia model of Haemophilus influenzae, Enterobacter cloacae, or extended-spectrum beta-lactamase (ESBL) nonproducing or producing Klebsiella pneumoniae pneumonia, we compared results of treatment with ceftobiprole medocaril (71 mg/kg, sc, qid), ceftriaxone (50 mg/kg, im, bid), or cefepime (50 mg/kg, ip, q.i.d.). Results were expressed as median and 25th to 75th percentile log10 colony forming units per gram of lung tissue. Ceftobiprole, ceftriaxone, and cefepime were each more active than was no treatment and were equally active for treatment of experimental H. influenzae, E. cloacae, or ESBL-nonproducing K. pneumoniae pneumonia. For ESBL-producing K. pneumoniae, no differences were detected between no treatment and treatment with ceftobiprole, ceftriaxone, or cefepime. Ceftobiprole is active against H. influenzae, E. cloacae, and ESBL-nonproducing K. pneumoniae in an immunocompetent experimental murine pneumonia model.

Animals↗
PubMed2005

An Enterobacter cloacae toxin able to generate oxidative stress and to provoke dose-dependent lysis of leukocytes.

We investigated an Enterobacter cloacae strain exhibiting high hemolytic and leukotoxic activity. Monomeric and polymeric forms of the toxin showed similar effects on blood cells, although the polymer was more active than the monomer. Fluorescence microscopy revealed that both forms of the FITC-labeled toxin interacted with leukocytes, principally with neutrophils. Prelytic concentrations of polymeric and monomeric toxin significantly increased the production of reactive oxygen species (ROS) in neutrophils. Conversely, lytic concentrations of both toxin forms showed an increase followed by a decrease of ROS due to neutrophil damage. Monocytes did not show oxidative stress at all the toxin concentrations assayed. The toxin-neutrophil interaction at prelytic concentrations of toxin-stimulated ROS production and led to oxidative stress with subsequent cell death by apoptosis. However, high concentrations of E. cloacae toxin damaged leukocytes, producing lysis before the trigger of apoptosis, which suggests that the toxic effect is concentration dependent. The inhibition of oxidative stress observed with genistein and chloroquine suggests a potential involvement of the tyrosine kinase and nitric oxide synthesis pathways in E. cloacae toxin-mediated elevation of ROS.

Apoptosis↗
PubMed2005

Pore formation, polymerization, hemolytic and leukotoxic effects of a new Enterobacter cloacae toxin neutralized by antiserum.

A new toxin of Enterobacter cloacae was purified and studied by SDS-PAGE electrophoresis with the purpose of investigating its ability to generate polymers and their molecular mass. Monomer of 13.3 kDa and structures of multimeric mass were detected. The toxin of 66 kDa was the most abundant form of toxin. This polymer and the monomer were selected to examine blood cells damage. Membrane pores caused by both toxin forms seemed to be of similar dimension (estimated in 3.6 nm by experiments with osmotic protectors) and were able to lyse erythrocytes and leukocytes. The results obtained indicate that polymerization and pore formation are involved in the molecular events that participate in the cytotoxic effects of E. cloacae toxin. Immunization of rabbits with 13.3kDa toxin generated antibody response capable of inhibiting oxidative stress as well as hemolytic and leukotoxic effects. Immunoblotting indicated that monomer and polymer reacted with antihemolysin serum. The importance of E. cloacae toxin "in vivo" was studied in animals by means of assays performed in peritoneum of rats, inoculated with the hemolytic strain (C1) and a non-hemolytic variant (C4). Both strains stimulated infiltration of leukocytes in peritoneum, but C1 caused cell death and lysis wheras assays with C4 maintained the viability of leukocytes even within 5 h after extraction of samples.

Animals↗
PubMed2005

A toxaphene-degrading bacterium related to Enterobacter cloacae, strain D1 isolated from aged contaminated soil in Nicaragua.

Enterobacter sp. strain D1 is a facultative anaerobic, Gram-negative heterotrophic bacterium isolated from toxaphene-contaminated soil. This organism was identified and characterized through phylogenetic and taxonomic studies. Based on 16S rDNA analysis, the strain D1 was clustered closely with the species Enterobacter cloacae subsp. dissolvens (LMG 2683) and E. cloacae (ATCC 13047T). Strain D1 resembled these E. cloacae strains with respect to various biochemical and nutritional characteristics, but also exhibited differences. Moreover, strain D1 is able to grow and survive with toxaphene supplied in the medium in the range 3-96 mg/L. Amongst the chemical components of toxaphene, octachlorocamphenes, nonachlorobornanes and decachlorobornanes were seen to be rapidly metabolized, although levels of hexachlorocamphenes and heptachlorobornanes were found to be slowly degraded, and subsequently accumulated during the last stage of the cultivation.

Biodegradation, Environmental↗
PubMed1997

Total urogenital mobilization--an easier way to repair cloacas.

The surgical treatment of persistent cloaca is a serious challenge. The operation is technically difficult and the final results for urinary and fecal function are far from excellent. The repair of a cloaca includes, among other maneuvers, the separation of the vagina from the urinary tract. This step is a serious technical challenge and is very time consuming. Devascularization of these structures is the main source of complications such as urethro-vaginal fistula, vaginal stricture, and acquired vaginal atresia. To avoid these complications and to facilitate the cloacal repair, a new technical variation using total urogenital mobilization was performed in 11 patients. In this procedure, after the rectum is separated from the vagina, both the urethra and the vagina are mobilized together as a single unit. The surgical time spent during the reconstruction was reduced by approximately 70%. All patients recovered well from the operations and have been followed up for 1 to 14 months. The blood supply of the vagina and urethra in all cases remained excellent. No patient developed urethrovaginal fistula, vaginal stricture, or acquired vaginal atresia. The cosmetic appearance in these patients is superior to the one achieved with previous techniques. Although this maneuver may not render better urinary or fecal control, the urethra is more accessible for catheterization. These preliminary results suggest that the total urogenital mobilization maneuver provides a definite technical advance in the repair of cloaca malformations.

Child, Preschool↗