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PubMed1998

Molecular analysis of glycopeptide-resistant Enterococcus faecium isolates collected from Michigan hospitals over a 6-year period.

The purpose of this study was to evaluate the molecular relatedness of clinical isolates of glycopeptide-resistant Enterococcus faecium isolates collected from hospitals in Michigan. A total of 379 isolates used in this study were all vancomycin-resistant E. faecium isolates collected from 28 hospitals and three extended-care facilities over a 6-year period from 1991 to 1996. For the 379 isolates, there were 73 pulsed-field gel electrophoresis (PFGE) strain types. Within strain types, there were as many as six restriction fragment differences. Most isolates (70%) belonged to six strain types, which were designated M1 (36%), M2 (3%), M3 (18%), M4 (6%), M10 (4%), and M11 (3%). PFGE strain M1 was cultured from 135 patients in 13 hospitals during the period 1993 to 1996. Strain type M2 was cultured from 11 patients in two hospitals during the period 1991 to 1992 and was not observed after 1992. Strain type M3 was cultured from 70 patients in 10 hospitals during the period of 1994 to 1996. Both M4 and M10 were cultured from 23 patients in three hospitals and from 15 patients in two hospitals, respectively, during 1995 to 1996. M11 was cultured from 13 patients in four hospitals during 1996. A total of 23 of 28 hospitals had evidence of clonal dissemination of some isolates. Plasmid content and hybridization analysis done on 103 isolates from one hospital and two affiliated extended-care facilities indicated that the strains contained from one to eight plasmids. Mating experiments indicated transfer of vancomycin resistance from 94 of these isolates into plasmid-free E. faecium GE-1 at transfer frequencies of <10(-9) to 10(-4). Gentamicin resistance and erythromycin resistance were cotransferred at various frequencies. A probe for the vanA gene hybridized to the plasmids of 23 isolates and to the chromosomes of 72 isolates. A probe for the vanB gene hybridized to the chromosomes of 8 isolates. The results of this study suggest inter- and intrahospital dissemination of vancomycin-resistant E. faecium strains over a 6-year period in southeastern Michigan. The majority of isolates studied belonged to the same few PFGE strains, indicating that clonal dissemination was responsible for most of the spread of resistance that occurred.

Anti-Bacterial Agents↗

PubMedDate not supplied

Role of penicillin-binding protein 5 C-terminal amino acid substitutions in conferring ampicillin resistance in Norwegian clinical strains of Enterococcus faecium.

The importance of amino acid sequence differences in the C-terminal part and levels of mRNA expression of penicillin-binding protein 5 (PBP5) for ampicillin resistance in Enterococcus faecium was investigated. Seventeen isolates from Norwegian hospitalized patients (ampicillin MIC 0.064->256 mg/L) with different C-terminal pbp5 DNA sequences encoding 11 different amino acid sequences were analyzed with a 14C-radiolabeled penicillin- binding assay to PBP5 and with real-time PCR quantification of pbp5 mRNA expression. Using multiple logistic regression analysis the amino acid substitution Met 485 was linked to ampicillin MIC and levels of 14C-radiolabeled penicillin bound to PBP5; however, there were isolates with identical PBP5 alleles and different ampicillin MICs. There was no relation between the quantity of pbp5 mRNA transcripts and ampicillin resistance. The results cannot explain ampicillin resistance in Norwegian clinical strains of E. faecium and indicate that other factors besides the properties of the C-terminal part of PBP5 are most likely involved.

Amino Acid Substitution↗

PubMed2006

Nosocomial spread of Enterococcus faecium resistant to vancomycin and linezolid in a tertiary care medical center.

In May 2004 our institution encountered its first clinical isolate of linezolid-resistant, vancomycin-resistant Enterococcus faecium (LRVRE). Between October 2004 and July 2005, 40 patients from whom LRVRE organisms were recovered in clinical specimens were characterized. Epidemiologic investigation and pulsed-field gel electrophoresis patterns indicated a clonal outbreak related to nosocomial spread.

Academic Medical Centers↗

PubMed2003

Clinical isolates of Enterococcus faecium exhibit strain-specific collagen binding mediated by Acm, a new member of the MSCRAMM family.

A collagen-binding adhesin of Enterococcus faecium, Acm, was identified. Acm shows 62% similarity to the Staphylococcus aureus collagen adhesin Cna over the entire protein and is more similar to Cna (60% and 75% similarity with Cna A and B domains respectively) than to the Enterococcus faecalis collagen-binding adhesin, Ace, which shares homology with Acm only in the A domain. Despite the detection of acm in 32 out of 32 E. faecium isolates, only 11 of these (all clinical isolates, including four vancomycin-resistant endocarditis isolates and seven other isolates) exhibited binding to collagen type I (CI). Although acm from three CI-binding vancomycin-resistant E. faecium clinical isolates showed 100% identity, analysis of acm genes and their promoter regions from six non-CI-binding strains identified deletions or mutations that introduced stop codons and/or IS elements within the gene or the promoter region in five out of six strains, suggesting that the presence of an intact functional acm gene is necessary for binding of E. faecium strains to CI. Recombinant Acm A domain showed specific and concentration-dependent binding to collagen, and this protein competed with E. faecium binding to immobilized CI. Consistent with the adherence phenotype and sequence data, probing with Acm-specific IgGs purified from anti-recombinant Acm A polyclonal rabbit serum confirmed the surface expression of Acm in three out of three collagen-binding clinical isolates of E. faecium tested, but in none of the strains with a non-functional pseudo acm gene. Introduction of a functional acm gene into two non-CI-binding natural acm mutant strains conferred a CI-binding phenotype, further confirming that native Acm is sufficient for the binding of E. faecium to CI. These results demonstrate that acm, which encodes a potential virulence factor, is functional only in certain infection-derived clinical isolates of E. faecium, and suggest that Acm is the primary adhesin responsible for the ability of E. faecium to bind collagen.

Adhesins, Bacterial↗

PubMed1995

Conjugative mobilization of a vancomycin resistance plasmid by a putative Enterococcus faecium sex pheromone response plasmid.

Recent epidemiological evidence suggests that horizontal gene transfer may be an important mechanism for dissemination of vancomycin resistance. A filter mating survey of 21 VanA Enterococcus faecium isolates from The New York Hospital showed that 14 of these isolates transferred vancomycin resistance (Vmr) to the plasmid-free reference strain Enterococcus faecalis JH2-2. One isolate, E. faecium R7, was selected for further study based on its ability to transfer Vmr to strain JH2-2 in liquid culture. Analysis of the plasmid content of transconjugants revealed three general classes. The predominant class (28 of 47 transconjugants) contained two separate plasmids: pHKK702 and pHKK703. pHKK702 is a 41-kb plasmid that contains an element indistinguishable from the Vmr transposon Tn1546 and an element that hybridizes with an ermB probe from the Staphylococcus aureus erythromycin resistance transposon Tn551. pHKK703 is a 55-kb plasmid that hybridizes with probes for the sex pheromone response genes prgA, prgB, and prgX derived from the E. faecalis plasmid pCF10. The second group of transconjugants (18 of 47) contained various recombinant forms of pHKK702 and pHKK703, whereas a third transconjugant class contained only pHKK702 (1 of 47). Transconjugants that contained both pHKK702 and pHKK703 were able to efficiently transfer Vmr to recipient strains in broth or on filters. However, no transfer of Vmr was detected using the donor containing only pHKK702. The transfer of Vmr from the recombination-deficient derivative of E. faecalis JH2-2 [strain UV202(pHKK702, pHKK703)] was reduced 600-fold compared to that of JH2-2(pHKK702, pHKK703). We propose that pHKK703 functions as an E. faecium sex pheromone response plasmid that conjugatively mobilizes pHKK702, and that a major pathway for this mobilization may require donor-mediated recombination proficiency. This report provides the first example in which a plasmid containing a Tn1546-related element is conjugatively mobilized.

Anti-Bacterial Agents↗

PubMed2004

Toxicity evaluation for an Enterococcus faecium strain TM39 in vitro and in vivo.

Previously, we have screened lactic acid bacteria (LAB) strains from infant feces and evaluated their functional properties. We found a strain of Enterococcus faecium termed as TM39 which is acid and bile tolerant, able to adhere to the intestinal epithelium and with antagonistic activity against Helicobacter pylori. In this study, we demonstrate that strain TM39 is not vancomycin resistant, not invasive to human gastric carcinoma cell line TSGH 9201 and human intestinal epithelial cell line Int-407 in vitro. In addition, we have conducted the in vivo study to evaluate the toxicity of this E. faecium strain TM39 in Wistar rats. For such study, cells of strain TM39 were daily oral administrated with dose of 1 x 10(12), 5 x 10(11) and 2 x 10(10) CFU/kg of body weight, respectively, to the rats for 28 consecutive days. There were no adverse effects on the general condition, behavior, growth, feed and water consumption, hematology, clinical chemistry values, organ weights and histopathologic analysis of the rats. Results of this study demonstrate that consumption of strain E. faecium TM39, even in large quantities, is not associated with any obvious signs of toxicity in Wistar rats.

Animals↗

PubMed2002

Comparison of an automated ribotyping system to restriction endonuclease analysis and pulsed-field gel electrophoresis for differentiating vancomycin-resistant Enterococcus faecium isolates.

The RiboPrinter Microbial Characterization System was compared with pulsed-field gel electrophoresis (PFGE), restriction endonuclease analysis (REA), and epidemiological data for typing 45 vancomycin-resistant Enterococcus faecium (VRE) isolates. In 21 clinically related isolates, 90 to 100% were similar by PFGE and REA, but only 57% were similar by the RiboPrinter. In another eight clinically related isolates, three isolates similar by PFGE and REA were all unique by the RiboPrinter. In contrast, in 16 clinically unrelated isolates, the predominant RiboPrinter ribotype represented 50% of the strains, while the largest PFGE and REA clones represented less than 19% of the strains. These data suggest that the RiboPrinter is not reliable for VRE investigation.

Automation↗

PubMed1999

Glycopeptide resistance in Enterococcus faecium from broilers and pigs following discontinued use of avoparcin.

The use of the glycopeptide growth promoter avoparcin was discontinued in Denmark in 1995 following concerns that vancomycin-resistant Enterococcus faecium occurring as a result of its use could be transferred to humans via food. The present study is an analysis of results obtained by the continuous surveillance of an antimicrobial resistance in Denmark (DANMAP) with the aim of determining the effect of the ban on the occurrence of glycopeptide resistance among E. faecium isolated from broilers and pigs. Among isolates from broilers, the proportion that were resistant to glycopeptides has shown a statistically high significant decline between the end of 1995 and the first half of 1998, whereas in pigs the ban appears to have no such effect. One possible explanation is that the broiler industry generally uses all in-all out production compared with continuous production in pig herds. Alternatively, the results indicate that the different outcomes may result from different co-selection patterns in pigs and broilers. In pigs, the antimicrobials most commonly used favored co-selection of glycopeptide resistant strains of E. faecium while in broilers the antimicrobials most widely used selected for glycopeptide-susceptible strains. The results show that intervention to reduce antimicrobial resistance may not always be effective and preventing resistance problems therefore becomes essential.

Animals↗

PubMed2001

Joint effect of nisin, CO2, and EDTA on the survival of Pseudomonas aeruginosa and Enterococcus faecium in a food model system.

A study on the joint effect of either nisin or Nisaplin, headspace CO2 levels, and EDTA on the survival of Pseudomonas aeruginosa and Enterococcus faecium was carried out in a water-soluble fish muscle extract at 3 degrees C using a second-order rotatable factorial design. E. faecium was completely deactivated by all processing after 2 days of storage. In contrast, P. aeruginosa was much less susceptible to treatments, and cell death was satisfactorily described by two models. Nisin increased cell death, whereas Nisaplin (commercial form of nisin) was not suitable, as it caused undesirable interference, presumably due to its co-compounds. Interactions between Nisaplin or nisin and either EDTA or CO2 were found to be nonstatistically significant. Factors that could account for this unexpected lack of synergism are discussed. However, a statistically significant positive interaction was found between CO2 and EDTA. This finding could allow CO2 levels to be decreased and hence to reduce the main disadvantages of CO2 application, namely, exudation and acidification.

Animals↗

PubMed2005

Screening of virulence determinants in Enterococcus faecium strains isolated from breast milk.

In a previous study, the authors isolated lactic acid bacteria from breast milk of healthy mothers. Since some of the identified isolates belonged to the species Enterococcus faecium, the objective of this work was to evaluate their safety. The enterococcal strains were screened by polymerase chain reaction (PCR) and Southern hybridization for the presence of virulence determinants. The potential of the strains to acquire plasmids by conjugation was investigated by screening for genes involved in conjugation processes. Parallel, phenotypic assays were performed. Presence of genes conferring resistance to vancomycin was assessed by PCR. PCR amplifications and Southern hybridizations revealed that all the strains were clear of the majority of potential virulence determinants. None of the strains showed gelatinase activity, hemolysin production, or aggregation phenotype, and none carried the vanA or vanB genes. These findings suggest that milk of healthy mothers may be a source of avirulent E faecium isolates to the newborns.

Base Sequence↗

PubMed1993

Constitutively vancomycin-resistant Enterococcus faecium resistant to synergistic beta-lactam combinations.

Vancomycin resistance among enterococci has recently been recognized. Synergy between vancomycin and penicillin has been shown in vitro for isolates of Enterococcus faecium resistant to both of these antibiotics. We describe three isolates of vancomycin-resistant E. faecium which demonstrate unique phenotypic characteristics. The isolates exhibited high-level resistance to both vancomycin and teicoplanin, consistent with the VanA phenotype. However, resistance in these isolates could not be induced or cured, and mating experiments failed to detect a transfer of resistance. The combination of vancomycin and penicillin did not significantly change the MIC of penicillin for any of the three isolates. Immunoblotting with polyclonal anti-VanB antibody showed no reaction with the cellular proteins of these strains. Probing with a vanA oligonucleotide revealed hybridization with chromosomal but not plasmid DNA. The mechanism of constitutive resistance of those strains remains unclear. A second mutational change, perhaps involving PBP 5, may explain the presence of resistance to synergistic combination penicillin-vancomycin therapy. In vitro evaluation of penicillin-vancomycin should be carried out in all clinical cases where this therapeutic regimen is being considered.

Adult↗

PubMed2005

High rate of resistance to quinupristin-dalfopristin in Enterococcus faecium clinical isolates from Korea.

We tested the in vitro susceptibilities of 603 enterococcal isolates from eight tertiary-care hospitals in Korea. The quinupristin-dalfopristin resistance rate in Enterococcus faecium was very high (25 isolates, 10.0%). It was suggested that both clonal spread and the sporadic emergence of quinupristin-dalfopristin-resistant isolates may explain the high prevalence of quinupristin-dalfopristin resistance in Korea.

Anti-Bacterial Agents↗

PubMed2005

Effects of zinc oxide and Enterococcus faecium SF68 dietary supplementation on the performance, intestinal microbiota and immune status of weaned piglets.

The objective of this study was to determine the effects of zinc oxide (ZnO) and the probiotic Enterococcus faecium SF68 (Cylactin) dietary supplementation on the performance, intestinal microbiota and immune parameters of the weaned piglet reared under commercial conditions. The diets were devoid of antibiotic growth promoters (AGP). Two hundred and eight crossbred piglets were allocated to a 2 x 2 factorial experiment involving two levels of zinc oxide supplementation (0 or 3100 mg ZnO/kg feed), and two levels of E. faecium SF68 supplementation (0 or 1.4 x 10(9)CFU/kg feed (Cylactin ME10)). The diets were offered ad libitum for 20 days post-weaning. Piglet performance was assessed by calculating average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR) on a pen basis. In addition, components of the distal ileal digesta, tissue-associated and mesenteric lymph node (MLN) bacterial populations were enumerated and serum immunoglobulin G (IgG) and intestinal immunoglobulin A (IgA) concentrations were determined on days 6 and 20 post-weaning. Regression analysis was used to determine the relationship between the bacterial populations at the different sites. Supplementation of the post-weaning diet with either ZnO or E. faecium SF68 did not affect piglet performance. E. faecium SF68 did not affect gastrointestinal bacterial populations but did tend to reduce serum IgG (P<0.1) on day 20. Zinc oxide reduced anaerobic (P<0.05) and tended to decrease lactic acid (P<0.1) bacterial translocation to the MLN, and tended to increase intestinal IgA concentration (P<0.1) on day 20. Generally, luminal bacterial populations were found to be poor predictors of tissue-associated or MLN populations. ZnO and E. faecium SF68 dietary supplementation were ineffective under these trial conditions. Further investigations into the possible immunomodulator role of dietary ZnO are warranted.

Animals↗

PubMed1993

The activity of daptomycin on Enterococcus faecium protoplasts: indirect evidence supporting a novel mode of action on lipoteichoic acid synthesis.

The effect of daptomycin, an acidic lipopeptide antibiotic active against Gram-positives, was studied in Enterococcus faecium protoplasts. This antibiotic killed 99% of the protoplasts within 60 minutes of treatment, while vancomycin was ineffective, thus excluding peptidoglycan synthesis as the only target of the action of daptomycin. As previously seen with whole cells, in protoplasts lipoteichoic acid synthesis was the earliest and most strongly inhibited among types of macro-molecular synthesis. Radioactive daptomycin tightly bound only to the cytoplasmic membrane, in which the enzymes involved in lipoteichoic acid synthesis are located. These conclusions strongly support our previous proposal that daptomycin, though active against peptidoglycan synthesis, primarily inhibits lipoteichoic acid synthesis.

Anti-Bacterial Agents↗

PubMed1995

Induction of vancomycin resistance in Enterococcus faecium by non-glycopeptide antibiotics.

Bacitracin and other antibiotics that inhibit late stages in peptidoglycan biosynthesis induce vancomycin resistance in a high-level, inducibly vancomycin-resistant strain of Enterococcus faecium. Exposure to bacitracin led to synthesis of the lactate-containing UDP-MurNAc-pentadepsipeptide precursor required for vancomycin resistance. These findings indicate that inhibition of peptidoglycan biosynthesis can lead to induction of vancomycin resistance and raise the possibility that multiple signals may serve to induce resistance.

Anti-Bacterial Agents↗

PubMed2003

Linezolid-resistant, vancomycin-resistant Enterococcus faecium infection in patients without prior exposure to linezolid.

We describe 2 patients without prior exposure to linezolid who were infected with closely related strains of linezolid- and vancomycin-resistant Enterococcus faecium (LRVREF) that may have been hospital acquired. Polymerase chain reaction amplification of the domain V region of the 23S ribosomal RNA gene demonstrated the presence of the G2576U mutation previously reported to be associated with linezolid resistance. Nosocomial transmission of LRVREF is an ominous sign and underscores the importance of meticulous infection-control measures.

Acetamides↗

PubMed2004

Influence of diet containing lyophilized Enterococcus faecium M-74 with organic selenium on tumor incidence in Apc1638N mice.

The aim of the present study was to test the effect of long-term application of diet containing Enterococcus faecium M-74 with organic selenium on tumor induction in transgenic mice carrying mutation in Apc gene. Heterozygosity for the Apc1638N mutation in mice causes development of small intestine and gastric tumors. Feeding of Apc1638N transgenic mice with enriched diet with probiotic components during 8 months have shown a minor therapeutic effect on the clinical manifestations in small intestine in comparison with control group.

Adenocarcinoma↗

PubMed2002

Molecular typing and antimicrobial susceptibility of vancomycin-resistant Enterococcus faecium in Brazil.

OBJECTIVES: To characterize vancomycin-resistant enterococci (VRE) isolates and to evaluate the mode of dissemination of this pathogen in Brazil. DESIGN: We collected 22 vancomycin-resistant Enterococcus faecium isolates from 6 medical centers in Sao Paulo, Brazil, and 1 isolate from a medical center in Curitiba, Brazil. PARTICIPANTS: All Brazilian hospitals that had identified vancomycin-resistant E. faecium up to the beginning of this study (late 1999) contributed isolates to the study. METHODS: The isolates were susceptibility tested using the broth microdilution method and the E-test. The presence of vancomycin resistance genes (vanA, vanB, vanC1, vanC2-3, and vanD) was evaluated by polymerase chain reaction; molecular typing was performed by pulsed-field gel electrophoresis (PFGE). RESULTS: The vanA gene was demonstrated in all vancomycin-resistant E. faecium, except for 1 isolate. None of the vancomycin resistance genes cited above was detected in the isolate from Curitiba, which was the first vancomycin-resistant E. faecium described in Brazil. All isolates were resistant to ampicillin and teicoplanin. The main clone remains susceptible to doxycycline and chloramphenicol, but intermediate to quinupristin-dalfopristin. PFGE analysis demonstrated 7 major PFGE patterns. A unique PFGE pattern with 4 subtypes was detected in 17 isolates from 4 different hospitals. CONCLUSION: The results of our study indicate the occurrence of intra- and interhospital dissemination of VRE in Sao Paulo, Brazil.

Brazil↗