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Diversity of aminoglycoside resistance in Enterobacter cloacae in Greece.

Ninety Enterobacter cloacae strains isolated from 12 Greek hospitals were examined in terms of epidemiological types and resistance mechanisms. Using O serotyping 69% of the strains were assigned to a specific serotype and overall 16 different serotypes were identified. The combination of serotyping, phagetyping and biotyping efficiently discriminated most of the strains, indicating that single epidemic strains were not prevalent, although serotypes 3, 7, and group II predominated. Eight representative strains, all resistant to gentamicin, tobramycin, amikacin and netilmicin, were further examined for transferability and mechanisms of resistance. Aminoglycoside resistance was found to be transferable in most strains, and 13 R plasmids of 40-120 MDa molecular weight were detected. The enzymes detected consisted of three enzymes active against gentamicin [ANT(2h'), AAC(3)-I and AAC(3)-V]; three active against tobramycin [ANT(2"), AAC(3)-V and AAC(6')-I]; two active against netilmicin [AAC(3)-V and AAC(6')-I]; and one active against amikacin [AAC(6')-I]. APH(3') and ANT (3"), which modify neomycin and streptomycin plus spectinomycin respectively, were also found. Overall up to five aminoglycoside modifying enzymes were detected on the same R plasmid, AAC(6')-I plus ANT(2") being the most prevalent. The high incidence of multiresistance in Enterobacter cloacae and the fact that resistance is due to enzymatic inactivation of the antibiotics, indicate that in Greece this species might act as a gene pool for the spread of resistance to other bacteria of clinical relevance.

Aminoglycosides↗

Type I beta-lactamases of Enterobacter cloacae and resistance to beta-lactam antibiotics.

The interaction of type-I beta-lactamases from Enterobacter cloacae with diverse beta-lactam compounds was examined. The ability of penicillin and cefoxitin to induce beta-lactamase production in this strain was assessed. The effect of beta-lactamase inhibitors was measured on beta-lactamase extracts and on intact cells. E. cloacae 78 strain is a stably derepressed mutant showing limited susceptibility to a number of antibiotics except imipenem. Imipenem would therefore be the appropriate choice for therapy of infections caused by stably derepressed mutants of Enterobacter sp. producing type-I beta-lactamases.

Anti-Bacterial Agents↗

Effects of carbon source and Vitreoscilla hemoglobin (VHb) on the production of beta-galactosidase in Enterobacter aerogenes.

At fixed concentration (0.5%), lactose and galactose acted as inducers while glucose and other tested carbon sugars showed repression effects on beta-galactosidase production in Enterobacter aerogenes strain. The expression of Vitreoscilla hemoglobin gene (vgb) in this bacterial strain managed to overcome the repression effects as well as improving the induction of beta-galactosidase formation by carbon sources. In parallel, the bacterial O(2) consumption was increased correspondingly to the vgb induction of beta-galactosidase synthesis. When Enterobacter aerogenes strains were grown at the incubation temperature 42 degrees C, about 5-fold higher enzyme productivity was obtained than with a similar incubation at 37 degrees C. The bacterial growth expressed as biomass yield had a different optimum temperature and was not influenced to the same extent by variations in the carbon sources. These data are discussed in terms of proposed enhancement in beta-galactosidase productivity by vgb expression as well as its significance to improve the technology of whey processing.

Bacterial Proteins↗

Serological cross-reactivity of environmental isolates of Enterobacter, Escherichia, Stenotrophomonas, and Aerococcus with Shigella spp.-specific antisera.

Using protocols designed for the isolation of Shigella from environmental freshwater samples from different regions of Bangladesh, 11 bacterial strains giving rise to Shigella-like colonies on selective agar plates and showing serological cross-reaction with Shigella-specific antisera were isolated. Phylogenetic analyses revealed that three of the isolates were most closely related to Escherichia coli, four to Enterobacter sp., two to Stenotrophomonas, and two isolates belonged to the Gram-positive genus Aerococcus. The isolates cross-reacted with six different serotypes of Shigella and were, in each case, highly type-specific. Two of the isolates belonging to the Enterobacter and Escherichia genera gave extremely strong cross-reactivity with Shigella dysenteriae and Shigella boydii antisera, respectively. The Aerococcus isolates gave relatively weak but significant cross-reactions with S. dysenteriae. Western blot analysis revealed that a number of antigens from the isolates cross-react with Shigella spp. The results indicate that important Shigella spp. surface antigens are shared by a number of environmental bacteria, which have implications for the use of serological methods in attempts for the detection and recovery of Shigella from aquatic environments.

Antigens, Bacterial↗

Cluster of neonatal infections in Jerusalem due to unusual biochemical variant of Enterobacter sakazakii.

Reported here is a cluster of infections due to a nitrate-negative variant of Enterobacter sakazakii, which occurred among premature neonates at the Hadassah Hospital, Mount Scopus, Jerusalem, in December 1999-January 2000. Pulsed-field gel electrophoresis showed cluster isolates to be identical but unrelated to previous systemic isolates recovered in 1993 and 1998. The organism was not isolated from infant formula powder, but it was recovered from prepared formula and from a kitchen blender. Elimination of the environmental focus, a change to factory-prepared infant formula, and isolation of affected infants terminated the event. Faecal carriage of Enterobacter sakazakii was observed for up to 18 weeks, emphasising the potential for cross-infection.

Bacteremia↗

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↗

Immunological relationship among glyceraldehyde-3-phosphate dehydrogenases in the genera Enterobacter and Escherichia.

The comparative immunological study of glyceraldehyde-3-phosphate dehydrogenase (G-3-PDH) among Enterobacteriaceae carried out with an anti-Enterobacter cloacae G-3-PDH serum pointed out the large heterogeneity of the genera Enterobacter and Escherichia. The use of two-dimensional maps integrating our new data and previously acquired quantitative data confirmed these results.

Enterobacter↗

Resistance to imipenem, cefepime, and cefpirome associated with mutation in Omp36 osmoporin of Enterobacter aerogenes.

Enterobacter aerogenes develops increased multidrug resistance via a functional alteration of outer-membrane permeability associated with a decrease in porin function. We have sequenced the gene coding the major porin of Enterobacter aerogenes, omp36. The sequence shows a high similarity with the Klebsiella pneumoniae ompK36 gene and is closely related to the enterobacterial OmpC family. Sequence analysis of several Omp36 issued from clinical strains indicated variability in putative cell-surface exposed domains. Interestingly, substitution Gly112Asp was observed in the conserved eyelet L3 region of the porin produced by two strains, C and 3. This substitution is associated with a high general beta-lactam resistance observed in these isolates and with alteration of pore properties previously described in strain 3 porin [Mol. Microbiol. 41 (2001) 189]. This is the first genetic identification of impermeability-mediated resistance to beta-lactams in various clinical E. aerogenes strains.

Bacterial Outer Membrane Proteins↗

A solvent tolerant isolate of Enterobacter aerogenes.

A solvent tolerant strain of Enterobacter aerogenes was isolated from soil by cyclohexane enrichment. Presence of cyclohexane (20%) in culture media prolonged the lag phase and caused reduction in biomass. Transmission electron micrographs showed convoluted cell membrane and accumulation of solvent in case of the cells grown in cyclohexane. The Enterobacter isolate was able to grow in the range of organic solvents having log P above 3.2 and also in presence of mercury, thus showing potential for treatment of solvent rich wastes.

Cyclohexanes↗

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↗

Comparative susceptibilities of Klebsiella species, Enterobacter species, and Pseudomonas aeruginosa to 11 antimicrobial agents in a tertiary-care university hospital.

The in vitro activity of cefepime was compared versus that of 10 antimicrobial agents commonly used in the treatment of serious infections caused by common aerobic gram-negative bacteria: aztreonam, cefoperazone, ceftazidime, ceftriaxone, ciprofloxacin, gentamicin, imipenem, piperacillin, ticarcillin-clavulanic acid, and tobramycin. We tested 30 clinical isolates representing a cross section of Klebsiella and Enterobacter species and Pseudomonas aeruginosa collected at our tertiary-care university hospital. The most potent beta-lactams were imipenem and cefepime, which demonstrated significant activity against the majority of strains in all 3 genera of bacteria tested, as did ciprofloxacin and tobramycin. Ceftazidime was active against Pseudomonas aeruginosa but was less potent against Klebsiella and Enterobacter spp. Cefoperazone and ceftriaxone were less active than ceftazidime against Pseudomonas aeruginosa. Cefepime was found to be highly active against many resistant organisms that traditionally have been difficult to treat.

Anti-Bacterial Agents↗

An outbreak of Enterobacter cloacae associated with contamination of a blood gas machine.

Over a 3-month period, five cases of Enterobacter cloacae bacteraemia occurred on our neonatal unit. In at least three of these, isolation of the organism coincided with clinical deterioration and evidence of sepsis. In one case, the same strain was isolated from an abscess on the neonate's forearm. The isolates had identical sensitivity patterns being resistant to all beta-lactams tested except imipenem. The extended time course of the infections made cross-infection an unlikely explanation. Moreover, close questioning of the staff and observation of their practices with regard to blood culture collection, failed to reveal any likely mechanism for pseudobacteraemia. On extensive investigation of the environment to try to identify a potential source of the organism, a strain of Enterobacter cloacae, was isolated from the probe of the blood gas machine and the probe cover. No other environmental samples were found to harbour the organism. Subsequent typing procedures showed the blood gas isolate to be indistinguishable from the clinical isolates. Five neonates were successfully treated with imipenem and gentamicin. The exact mechanism whereby these bacteraemias occurred remains obscure. In one case, the baby had positive blood cultures within 2 h of being on the unit and contamination of the blood culture bottle by the doctor taking the culture was suspected. Most of the episodes, however, appeared clinically to be genuine septicaemias. When vigorous infection control procedures were instituted to prevent staff acquisition of the organism from the machine, cases on the unit ceased.

Anti-Bacterial Agents↗

Enterobacter sakazakii: a review.

Enterobacter sakazakii, previously referred to as a yellow-pigmented Enterobacter cloacae was designated as a unique species in 1980. This reclassification was based on differences from E. cloacae in DNA relatedness, pigment production and biochemical reactions. E. sakazakii has been implicated in a severe form of neonatal meningitis. Although studies have failed to identify an environmental source for the organism, dried-infant formula has been implicated in both outbreaks and sporadic cases of E. sakazakii meningitis. The high mortality rate (40-80%), the severity of the infection in infants, plus the scarcity of information on the ecology and pathogenicity of this organism warranted a review of the clinical and microbiological features of this putative foodborne pathogen.

Enterobacter↗

Detection of amp C in Enterobacter cloacae in China.

PCR amplification of 55 strains of Enterobacter cloacae indicated 51 of them had amp C structural gene verified by DNA sequence and Southern blotting. All PCR products were cleaved into 666- and 328-bp fragments by Kpn1 restriction enzyme. Imipenem was the most potent inducer for mRNA expression of amp C gene and beta-lactamase activity. The beta-Lactamase inhibitor R0481220 strongly inhibited Amp C beta-lactamases; 96.4% (53/55) of Enterobacter cloacae producing Amp C enzyme were susceptible to cefepime.

Bacterial Proteins↗

Natural antibiotic susceptibility of strains of the Enterobacter cloacae complex.

The natural susceptibility to 70 antibiotics of 104 strains of the Enterobacter cloacae complex was examined using a microdilution procedure in Isosensitest broth. One hundred and one clinical strains designated as 'E. cloacae' were identified as E. hormaechei (n=65), E. asburiae (n=20) and E. cloacae genospecies 1 or 2 (n=16). Apart from fosfomycin susceptibility, there were only minor differences in natural antibiotic susceptibility patterns. All species were naturally sensitive or naturally sensitive and intermediate to numerous beta-lactams (e.g. carbapenems, aztreonam, acylaminopenicillins, ticarcillin, and some 'modern' cephalosporins), quinolones, aminoglycosides, tetracyclines, antifolates, chloramphenicol, and nitrofurantoin. Uniform natural resistance was found in benzylpenicillin, oxacillin, amoxycillin, amoxycillin/clavulanate, cefoxitin, rifampicin, lincosamides, glycopeptides, streptogramins and fusidic acid. Enterobacter hormaechei was the species most susceptible to fosfomycin. beta-Lactam susceptibility patterns pointed to the presence of chromosomally-encoded AmpC enzymes in all taxa of the E. cloacae complex.

Anti-Bacterial Agents↗

Molecular epidemiology and antibiotic resistance of Enterobacter spp. from three distinct populations in Grampian, UK.

The distribution of Enterobacter spp. within the population of Aberdeen Royal Infirmary was compared with the outpatient population with regard to molecular epidemiology and antibiotic resistance. Enterobacter spp. from 60 patients and one environmental site were characterised as ITU, non ITU and outpatients' isolates. Thirty-five percent were blood culture isolates. Cefotaxime resistant strains in the hospital were frequent. Cefotaxime (64%) sensitive isolates were inducible for hyperproduction of Bush group 1 beta-lactamase. Isolates were further investigated by PFGE. Isolates (27%) were clonally related and typed in four clusters. Consecutive isolates were studied in selected patients showing minor genomic changes. One environmental isolate from a deep sink at ITU was related to a patient's isolate.

Anti-Bacterial Agents↗

Yersiniabactin and other siderophores produced by clinical isolates of Enterobacter spp. and Citrobacter spp.

We analyzed the ability of extraintestinal strains of Enterobacter spp. and Citrobacter spp. to employ different siderophore-mediated strategies of iron acquisition. All strains produced iron-chelating compounds. Cross-feeding assays indicated that most isolates of both Enterobacter spp. and Citrobacter spp. excreted catecholate siderophore enterobactin, less produced aerobactin, and single strains excreted hydroxamates different from aerobactin. Besides, we analyzed if the strains had the ability to produce the siderophore yersiniabactin coded by the Yersinia high-pathogenicity island (HPI). The presence of HPI genes was observed in single isolates of three species: E. cloaceae, E. aerogenes and C. koseri. A detailed polymerase chain reaction analysis revealed differences in the genetic organization of the HPIs; however, in a cross-feeding test we proved that yersiniabactin was produced and the island was functional.

Bacterial Proteins↗

Enterobacter sepsis in infants and children due to contaminated intravenous fluids.

Sixty-three cases of nosocomial sepsis occurring from April through October 1981, in a 500-bed pediatric hospital, were traced to bacterial contamination of intravenous fluid produced by a single manufacturer. Two species of uncommon blood stream pathogens, Enterobacter cloacae and Enterobacter agglomerans contaminated the fluid. Infections with these organisms might have contributed to the death of four patients; two who were immunosuppressed, one who was asplenic and one premature infant. Epidemiologic and laboratory investigations identified the site of contamination to be within the screw-caps of the bottles containing the intravenous fluid. Contamination occurred during insertion of the intravenous fluid administration set into the bottle. The "epidemic" terminated when the hospital discontinued the use of infusion fluids from that manufacturer. We conclude that intravenous fluids should be examined during outbreaks of nosocomial bacteremia due to unusual pathogens.

Adolescent↗