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[Multicenter trial of Enterococcus antibiotic susceptibility].

Antibiotic susceptibility of 446 Enterococcus isolates from 9 medical centres of Moscow and St. Petersburg was tested. Among the isolates 386 belonged to E.faecalis, 48 to E.faecium and 12 to the other species. All the isolates were susceptible to vancomycin. As for E.faecalis 84 and 85 per cent of the isolates were susceptible to ampicillin and ampicillin/sulbactam respectively (no production of beta-lactamases), the frequency of high resistance to aminoglycosides amounted to 44 per cent with respect to streptomycin and to 25 per cent with respect to gentamicin, 75 per cent of the isolates was susceptible to ciprofloxacin. As for E.faecium and the rare species of Enterococcus more than 70 per cent of the isolates was resistant to ampicillin and ampicillin/sulbactam, the frequency of high resistance to aminoglycosides exceeded 60 per cent, 17 and 25 per cent of the isolates were susceptible to ciprofloxacin.

Ampicillin↗

Investigation of an outbreak of vancomycin-resistant Enterococcus faecium in a low prevalence university hospital.

BACKGROUND: Until 1995, there were no cases of vancomycin resistant enterococcus (VRE) identified at our university hospital. From May 1995 to August 1996, we investigated a cluster of 10 cases of phenotypic class Van B Enterococcus faecium. METHODS: Patients were matched with controls who were on the same unit for at least 7 days prior to the case developing VRE. Control patients were age and sex matched if possible, and had duration of hospitalization at least as long as the number of days it took the patient to become VRE positive. We analyzed 16 independent risk factors using Epi-info version 6. Environmental cultures were obtained in the MICU where 5 of the patients were located. All 10 patient isolates and environmental isolates were analyzed by pulsed field gel electrophoresis (PFGE). RESULTS: PFGE confirmed the genetic relatedness of all 10 patient isolates and environmental isolates. The VRE-positive group was more likely to be immunosuppressed and to have exposure to 3 physicians. In the MICU, significant, P < 0.05) risk factors for VRE were higher Apache scores, location adjacent to a VRE case, duration of vancomycin and amino-glycoside use, duration of invasive catheter use, and diarrhea. Among the VRE-positive environmental cultures was a blood pressure cuff wash that was used on several patients. CONCLUSION: We hypothesize that a VRE strain was introduced into our hospital environment and was spread by personnel or contaminated equipment. As a consequence of this study, a hospital-wide VRE policy was implemented.

Adolescent↗

Vancomycin resistance is encoded on a pheromone response plasmid in Enterococcus faecium 228.

In Enterococcus faecium 228, vancomycin resistance is encoded on a 55-kilobase conjugative plasmid, pHKK100. This plasmid was transferred with high frequency into susceptible strains of Enterococcus faecalis and conferred responses to pheromones produced by E. faecalis and Streptococcus sanguis. pHKK100 is the first plasmid described that mediates both vancomycin resistance and pheromone response.

Conjugation, Genetic↗

Cloning and heterospecific expression of the resistance determinant vanA encoding high-level resistance to glycopeptides in Enterococcus faecium BM4147.

Fragments of plasmid pIP816, which confers high-level glycopeptide resistance in Enterococcus faecium BM4147, were cloned into a conjugative gram-negative-gram-positive shuttle vector. The resulting hybrids were transferred by conjugation from Escherichia coli to Enterococcus faecalis and Bacillus thuringiensis. A 4-kilobase EcoRI fragment from pIP816 was found to confer vancomycin resistance in these hosts but not in E. coli or Bacillus subtilis.

Cloning, Molecular↗

Tetracycline resistance heterogeneity in Enterococcus faecium.

The tetracycline (Tet) determinants, which encode resistance either to tetracyclines without minocycline (Tcr) or to tetracyclines including minocycline (Tcr-Mnr), of 30 wild-type clinical isolates of Enterococcus faecium were identified and localized. The Tet determinants were transferred by conjugation into a plasmid-free Enterococcus faecalis recipient at frequencies of 10(-6) to 10(-9) transconjugants per donor, as follows: Tcr, 6 strains; Tcr-Mnr, 14 strains; both Tcr and Tcr-Mnr, 6 strains; no detectable transfer, 4 strains. Classes L (Tcr phenotype) and M and O (Tcr-Mnr phenotype) of the Tet determinants were identified by DNA-DNA hybridization experiments. The Tet L determinant was plasmid-borne in 18 strains and was chromosomal in 2 strains. Tet M was chromosomal in 27 strains and plasmid-borne (pIP1534) in 1 strain; pIP1534 also carried Tet L. Tet M was located on Tn916-like elements in 22 strains and on a Tn916-modified element in 1 strain. Tet O was detected in only one strain in which it was plasmid-borne. Both Tet L and Tet M determinants were carried by 19 strains. One strain carried, in addition to chromosomal nonconjugative Tet L and Tet M determinants, a conjugative Tcr-Mnr marker which did not correspond to any Tet determinant tested in this study. These results attest to the genetic complexity of tetracycline resistance in E. faecium strains.

Chromosomes, Bacterial↗

Pfizer selective enterococcus agar overlay method for the enumeration of fecal streptococci by membrane filtration.

The use of Pfizer selective enterococcus (PSE) agar with the membrane filter technique for the enumeration of fecal streptococci is limited due to the inability of the characteristic black precipitate, indicative of esculin hydrolysis, to diffuse from the medium through the membrane. A modification of the membrane filter technique that consisted of placing the membrane on PSE agar and overlaying it with tempered PSE agar was evaluated by comparing recovery, selectivity, and other parameters with M-enterococcus and KF-streptococcus agars, two selective media routinely used with the membrane filter technique for the enumeration of fecal streptococci in water and wastewater. No statistically significant differences could be demonstrated in the recovery capabilities of the three media. Inasmuch as the PSE overlay technique requires only 24 h of incubation as opposed to 48 h for the other two media, this modification may have some merit in water pollution monitoring programs.

Agar↗

The life and times of the Enterococcus.

Enterococci are important human pathogens that are increasingly resistant to antimicrobial agents. These organisms were previously considered part of the genus Streptococcus but have recently been reclassified into their own genus, called Enterococcus. To date, 12 species pathogenic for humans have been described, including the most common human isolates, Enterococcus faecalis and E. faecium. Enterococci cause between 5 and 15% of cases of endocarditis, which is best treated by the combination of a cell wall-active agent (such as penicillin or vancomycin, neither of which alone is usually bactericidal) and an aminoglycoside to which the organism is not highly resistant; this characteristically results in a synergistic bactericidal effect. High-level resistance (MIC, greater than or equal to 2,000 micrograms/ml) to the aminoglycoside eliminates the expected bactericidal effect, and such resistance has now been described for all aminoglycosides. Enterococci can also cause urinary tract infections; intraabdominal, pelvic, and wound infections; superinfections (particularly in patients receiving expanded-spectrum cephalosporins); and bacteremias (often together with other organisms). They are now the third most common organism seen in nosocomial infections. For most of these infections, single-drug therapy, most often with penicillin, ampicillin, or vancomycin, is adequate. Enterococci have a large number of both inherent and acquired resistance traits, including resistance to cephalosporins, clindamycin, tetracycline, and penicillinase-resistant penicillins such as oxacillin, among others. The most recent resistance traits reported are penicillinase resistance (apparently acquired from staphylococci) and vancomycin resistance, both of which can be transferred to other enterococci. It appears likely that we will soon be faced with increasing numbers of enterococci for which there is no adequate therapy.

Drug Resistance, Microbial↗

Vancomycin-Resistant Enterococcus faecium: First Case in Brazil.

We report a fatal case of septicemia due to a vancomycin-resistant Enterococcus faecium in a 9 year old girl with aplastic anemia. The isolate was also resistant to ampicillin, teicoplanin, gentamicin (high level), and streptomycin (high level). We believe that this is the first case of vancomycin-resistant Enterococcus (VRE) reported from a clinical specimen in Brazil.

Journal Article↗

[Effectiveness of ClNa-esculin broth for the rapid identification of the Enterococcus genus].

The efficacy of the ClNa-esculine broth for the rapid identification (2 hours) of the species of the genus Enterococcus was evaluated and compared with the conventional biochemical methods requiring an incubation of 18-24 hours. Out of the 274 evaluated strains, 171 were positive for this investigation; 160 of them corresponded to Enterococcus spp (100%). The sensitivity of the test was 100%, the specificity 99.1%, the positive predictive value 99.3% and the negative predictive value 100%.

Culture Media↗

In vivo v in vitro susceptibility of enterococcus to trimethoprim-sulfamethoxazole. A pitfall.

Two patients with uncomplicated enterococcal urinary tract infections were treated with trimethoprim-sulfamethoxazole based on in vitro susceptibilities. Bacteremia developed in both patients and they recovered only after the cessation of trimethoprim-sulfamethoxazole administration and institution of therapy with penicillin G potassium or vancomycin hydrochloride plus streptomycin sulfate. Although the enterococcus may appear susceptible to trimethoprim-sulfamethoxazole in vitro, it escapes the antifolate activity of the drug in vivo by its unique ability to incorporate preformed exogenous folates. The practice by clinical microbiology laboratories of reporting the susceptibilities of the enterococcus to drugs other than the penicillins or vancomycin is misleading and potentially dangerous.

Drug Combinations↗

Identification of a novel insertion sequence in vanB2-containing Enterococcus faecium.

AIMS: The characterization of a novel insertion sequence (IS) in vanB2-containing Enterococcus faecium was conducted. METHODS AND RESULTS: Direct PCR amplification of ORFC region of Tn5382 from DNA extracted from vanB2-containing E. faecium, and sequence analysis were performed. A novel IS was identified. It is 1418 bp in length and contains one putative open reading frame that is similar to transposase. There exists inverted terminal repeats of 12 bp, but direct repeats are not present. According to high similarity to putative transposases of IS3 members, such as, IS150, IS861, IS1077 and IS911, we designated it ISEnfa3. SIGNIFICANCE AND IMPACT OF STUDY: Since ISEnfa3 was detected in all vanB2-containing strains examined so far, it could be used as a tool for epidemiological study.

Amino Acid Sequence↗

Mutations in ribosomal protein L16 and in 23S rRNA in Enterococcus strains for which evernimicin MICs differ.

Mutations in ribosomal protein L16 and in 23S rRNA were investigated in 22 Enterococcus strains of different species and for which the MICs of evernimicin differ (MICs, 0.023 to 16 micro g/ml). Amino acid changes (Arg56His, Ile52Thr, or Arg51His) in protein L16 were found in seven strains, and a nucleotide G2535A mutation in 23S rRNA was found in 1 strain among 13 for which the MICs are > or =1 micro g/ml.

Amino Acid Substitution↗

Multiple-antibiotic resistance of Enterococcus spp. isolated from commercial poultry production environments.

The potential impact of food animals in the production environment on the bacterial population as a result of antimicrobial drug use for growth enhancement continues to be a cause for concern. Enterococci from 82 farms within a poultry production region on the eastern seaboard were isolated to establish a baseline of susceptibility profiles for a number of antimicrobials used in production as well as clinical environments. Of the 541 isolates recovered, Enterococcus faecalis (53%) and E. faecium (31%) were the predominant species, while multiresistant antimicrobial phenotypes were observed among all species. The prevalence of resistance among isolates of E. faecalis was comparatively higher among lincosamide, macrolide, and tetracycline antimicrobials, while isolates of E. faecium were observed to be more frequently resistant to fluoroquinolones and penicillins. Notably, 63% of the E. faecium isolates were resistant to the streptogramin quinupristin-dalfopristin, while high-level gentamicin resistance was observed only among the E. faecalis population, of which 7% of the isolates were resistant. The primary observations are that enterococci can be frequently isolated from the poultry production environment and can be multiresistant to antimicrobials used in human medicine. The high frequency with which resistant enterococci are isolated from this environment suggests that these organisms might be useful as sentinels to monitor the development of resistance resulting from the usage of antimicrobial agents in animal production.

Animal Husbandry↗

[The trend and susceptibility to antibacterial agents of enterococcus species from urinary tract infections].

OBJECTIVE: To determine the extent of drug-resistance among Enterococcus species we investigated in vitro experiments. METHODS: Studies were carried out on pure cultured of enterococci isolated from 8,575 urine specimens between 1990 and 2002. We had determined test strains to three kinds of species, which posses the urinary pathogenesis. Both an EF-agars and an ADH decarboxylase test performed the identification and speciation of the strains of enterococci. In vitro drug-susceptibility tests of enterococci were performed against the following antibiotics: ampicillin (ABPC), cefpirome (CPR), cefozopran (CZOP), imipenem/cilastatin (IPM/CS), minocycline (MINO), levofloxacin (LVFX), vancomycin (VCM), sulfamethoxazone/trimethoprim (ST), by employing the method for dilution antimicrobial susceptibility tests for bacteria that grow aerobically recommended by Japan Society of Chemotherapy. These drug-susceptibilities were shown susceptible, intermittent and resistant in according to National Committee for Clinical Laboratory Standards (M100-S12). RESULTS: The most common species isolated was E. faecalis (84.4%), followed by E. faecium (9.9%) and E. avium (5.6%). In E. faecium and E. avium, the sensitivity to ABPC has tended to improve from 1999. This tendency inverse correlated to decreasing dosage of PCs. There was much difference of resistant rate to IPM/CS between each species, and no correlation to used dosage of CBPs. The rate of resistance to MINO did not change during this period. 60% of E. faecalis had sensitivity to LVFX and the rate did not change during this period. In E. faecium, whose resistant rate to LVFX was 90%, the sensitivity has been improved to over 25% from 2001. The improved tendency of E. faecium to LVFX has inverse proportion to decreasing dosage of NQs. With the exception of a little bit VRE (VCM resistant Enterococci), almost of them had sensitivity to VCM. CONCLUSION: The emergence of enterococci with alarming rates of resistance concomitantly to multi-drugs highlights the need for a more rational and restricted use of antimicrobials, in order to minimize the selection and spread of such strains. An early detection of these problem pathogens is also important for preventing any treatment failures.

Ampicillin Resistance↗

Characteristics of Enterococcus strains isolated in 1991-1992.

The paper presents the identification and resistance to antibiotics as well as the high-level resistance of 96 strains of enterococci isolated in 1991-1992 from patients admitted in the "Victor Babes" Hospital of Infectious Diseases. The identification of Enterococcus species using the Facklam-Collins schedule and several additional tests reveal 86.4% E. faecalis, 7.2% E. faecium, 4.1% E. gallinarum and 1.0% E. durans and 1% E. hirae. The high-level resistance to aminoglycosides was 69.8% for Streptomycin, 50%--Kanamycin and 21.8%--Gentamicin. The phenotype of resistance to Streptomycin, Kanamycin and Gentamicin was found in 19.8% of strains. 7.4% of enterococci had MIC values higher or equal to 32 mcg/ml to Penicillin and MIC > or = to 16 mcg/ml to Ampicillin. Six E. faecalis strains and two E. gallinarum strains were resistant to Vancomycin.

Drug Resistance, Microbial↗

Construction of plasmid vector for expression of bacteriocin N15-encoding gene and effect of engineered bacteria on Enterococcus faecalis.

A 6.09-kb plasmids vector pOri253 was constructed from the plasmid pIL253 (5.2 kb) and a 0.89-kb fragment of oriColE1 from pBluescript II KS. The bifunctional plasmid pOri253 conferred erythromycin resistance in both Escherichia coli and Enterococcus faecalis. It has unique sites for EcoRI, BamHI, SalI, and PstI derived from pIL253 and was lost at a low rate in E. faecalis JCM8726 when cultured in Man, Rogosa, & Sharpe broth without antibiotic. The lactococcal promoter P23 was inserted at one end of the pOri253 multicloning site. Gene expression was assessed by an entAI gene, which produced bacteriocin N15. The E. faecalis harboring constructed plasmid carrying P23 (pOrient23) had more antibacterial activity than parental E. faecalis JCM8726 and its clone harboring non-P23-containing plasmid (pOrient), as determined by means of an overlay method.

Bacteriocins↗

Characterization of bacteriocins from Enterococcus faecium with activity against Listeria monocytogenes.

Laboratory cultures and environmental isolates of bacteria were screened for antagonism towards Listeria monocytogenes using an agar spot test. Seven of the 163 strains that were tested, one Streptococcus bovis, one Enterococcus casseliflavus, two E. avium and three E. faecium, consistently displayed antilisterial activity. Cell-free, pH-neutralized supernatants prepared from the three E. faecium strains (JBL1061, JBL1083 and JBL1351) exhibited strong antilisterial activity against L. monocytogenes, and were subjected to more detailed analyses. The antagonistic factors produced by these three strains were sensitive to chloroform and several proteolytic enzymes, resistant to heat (121 degrees C, 20 min), and stable over a wide pH range (3.0-10.0). Moreover, they were listericidal without causing cell lysis. These data suggest that a bacteriocin(s) is involved in the inhibition of L. monocytogenes by E. faecium JBL1061, JBL1083 and JBL1351.

Bacteriocins↗

Modular design of the Enterococcus hirae muramidase-2 and Streptococcus faecalis autolysin.

The mature forms of the extracellular muramidase-2 of Enterococcus hirae and Streptococcus faecalis autolysin have very similar primary structures. Each consists of an active-site-containing N-terminal domain fused to a multiple-repeat C-terminal domain. Polypeptide segments occurring at equivalent places in these two bacterial wall lytic enzymes have homologues in two phage lysozymes and in three functionally unrelated proteins, illustrating the principle that protein molecules frequently are constructed from modules that are linked in a single polypeptide chain.

Amino Acid Sequence↗