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PubMed1997

Incidence and detection of multi-drug-resistant enterococci in Dublin hospitals.

In February 1994, an outbreak of vancomycin-resistant Enterococcus faecium (VREM) occurred in the oncology unit of a Dublin hospital. Between February and July 1994, VREM was isolated from 18 patients, one staff member and 14 environmental sites within the unit. The isolates also had high-level aminoglycoside and penicillin resistance. Three pulsed-field gel electrophoresis (PFGE) types were identified, two of them from multiple patients and environmental sites. Plasmid typing allowed subdivision of PFGE types. A retrospective study of enterococci isolated from blood cultures between January 1991 and January 1994 showed that, before the outbreak, fewer than 2% of isolates were vancomycin-resistant but that the incidence of high-level gentamicin resistance had increased from 17% to 60% and ampicillin resistance from 22% to 51%. Among clinically significant non-blood-culture enterococci isolated between September and December 1993, fewer than 1% were vancomycin-resistant, 13% were ampicillin-resistant and 44% highly gentamicin-resistant. None produced beta-lactamase. High-content gentamicin disks (120 micrograms) and low-content vancomycin disks (5 micrograms) allowed simple, reliable detection of resistant enterococci. MICs of vancomycin and teicoplanin determined by agar dilution and E-test agreed well, but values tended to be slightly lower by E-test.

Ampicillin Resistance↗
PubMed1975

Activity of amoxycillin against enterococci and synergism with aminoglycoside antibiotics.

Amoxycillin was more active in vitro than ampicillin or benzylpenicillin against clinical isolates of enterococci. All 55 strains tested were sensitive to the three penicillins but 15 strains showed a high level of resistance to streptomycin and two of these were also insensitive to kanamycin. All strains were sensitive to gentamicin, which was the most active of the aminoglycoside antibiotics. The penicillins showed pronounced bactericidal activity against the enterococci but failed to sterilise cultures of these organisms. Combinations of penicillins and aminoglycosides invariably produced synergistic bactericidal effects which resulted in sterilisation of cultures of these organisms. Combinations of penicillins and aminoglycosides invariably produced synergistic bactericidal effects which resulted in sterilisation of cultures of enterococci provided that the strain was sensitive to the aminoglycoside moiety of the antibiotic combination. Synergism was not observed with a combination of a penicillin and an aminoglycoside when the enterococcus was resistant to the aminoglycoside. The data reported suggest that amoxycillin may have certain advantages, compared with ampicillin or benzylpenicillin, for the treatment of enterococcal infections.

Amoxicillin↗
PubMed2005

Characterization of antibiotic resistance genes and virulence factors in faecal enterococci of wild animals in Portugal.

Antibiotic susceptibility was tested in 140 non-selected enterococci (73 Enterococcus faecalis, 45 E. faecium and 22 of other species) recovered from faecal samples of 77 wild animals in Portugal. Susceptibility testing for 11 antibiotics (vancomycin, teicoplanin, ampicillin, streptomycin, gentamicin, kanamycin, chloramphenicol, tetracycline, erythromycin, quinupristin-dalfopristin and ciprofloxacin) was determined by disk diffusion and agar dilution methods. Forty-four isolates (31.4%) showed susceptibility to all the antibiotics tested (5.5% of E. faecalis; 62.2% of E. faecium; and 78.6% of E. hirae). Neither ampicillin-resistance nor acquired-vancomycin-resistance was detected and 1.4% of the isolates showed high-level-resistance for gentamicin or streptomycin. Tetracycline and erythromycin resistances were shown in 28.6% and 20.1% of the isolates, respectively. Antibiotic resistance genes were studied by polymerase chain reaction (PCR) and sequencing and tet(M) + tet(L), erm(B) or aac(6')-aph(2'') genes were detected in most of tetracycline-, erythromycin- or gentamicin-resistant enterococci respectively. Genes encoding virulence factors were studied by PCR and a wide variety of virulence genes were detected in most of E. faecalis isolates but were rarely found in E. faecium and not detected in the other species. The prevalence of genes encoding virulence factors in E. faecalis was as follows: cpd (98.6%), gelE (75.3%), agg (30.1%), fsr (17.8%), ace (9.6%) and esp (4.1%). Low percentages of antibiotic resistance was found in the faecal enterococci of wild animals but a wide variety of virulence genes were detected among E. faecalis isolates although were rare in the other species.

Animals↗
PubMed1996

Reevaluation of contemporary laboratory methods for detection of antimicrobial resistance among enterococci.

OBJECTIVE: To evaluate broth microdilution, disk diffusion, Etest and Vitek Systems for susceptibility testing of enterococci. METHODS: Susceptibility testing of a panel of 149 enterococci (99 vancomycin-resistant) strains, using the study methods, was performed and the results compared. RESULTS: For vancomycin susceptibility testing, categorical agreement of disk diffusion, Etest and Vitek with the reference broth microdilution test was > 95%. For aminoglycoside and ampicillin testing, categorical agreement between Etest and Vitek was 98 to 100%. CONCLUSIONS: Disk diffusion, Etest and Vitek have acceptable performance for detection of vancomycin resistance of Van A and Van B phenotypes among enterococci.

Journal Article↗
PubMed1997

Comparison of agar-based media for primary isolation of glycopeptide-resistant enterococci.

OBJECTIVE: To compare four vancomycin-containing agar media for the isolation of glycopeptide-resistant enterococci (GRE) from clinical fecal specimens: kanamycin---aesculin---azide (KAA) agar; bile---aesculin---polymixin (BAP) agar; aztreonam---amphotericin blood (CBAA) agar; and neomycin blood (CBN) agar. METHODS: Fecal specimens from 125 patients were inoculated onto each medium. Media were examined for enterococci after incubation for up to 48 h. Enterococci were identified to species level, and glycopeptide phenotypes were determined by measuring minimum inhibitory concentrations of vancomycin and teicoplanin. RESULTS: GRE were isolated from 44/125 samples. Enterococcus faecalis and Enterococcus faecium isolates, expressing glycopeptide resistance of the VanA or VanB phenotypes, were recovered from 27/33 (82%) specimens on BAP medium, 26/33 (79%) on KAA medium, and 21/33 (64%) on CBN and CBAA media. Enterococcus gallinarum and Enterococcus casseliflavus isolates expressing low-level glycopeptide resistance (VanC phenotype) were recovered from 14/15 (93%) specimens on CBAA medium, 7/15 (47%) on KAA and CBN media, and 6/15 (40%) on BAP medium. CONCLUSIONS: The media tested in this study, with the exception of CBN medium, detected at least 75% of patients colonized by GRE. Further development of BAP, CBAA and KAA media is warranted to improve growth and selectivity.

Journal Article↗
PubMed1997

Colonization by vancomycin-resistant enterococci of the intestinal tract of patients in intensive care units from French general hospitals.

OBJECTIVE: To evaluate the prevalence of fecal carriage of vancomycin-resistant enterococci (VRE) by patients hospitalized in intensive care units from 24 French general hospitals. METHODS: Rectal swabs were obtained from 647 patients hospitalized in intensive care units during the month of June 1994 and plated on agar medium selective for vancomycin-resistant enterococci. The glycopeptide resistance phenotypes and genotypes of the enterococci detected were characterized. RESULTS: Thirty-two of 647 patients (4.9%) carried VRE. Thirteen strains (2%) were identified as Enterococcus faecium and 19 (2.9%) as Enterococcus gallinarum or Enterococcus casseliflavus. None of these strains was highly resistant to gentamicin. The E. gallinarum and E. casseliflavus strains contained the vanC1 and vanC2 genes, respectively. The E. faecium strains were highly resistant to vancomycin and teicoplanin and carried the vanA gene. No infection due to VRE was observed during the study period. Pulsed-field gel analysis of total DNA following digestion with SmaI or KspI from 13 VanA-type E. faecium strains revealed intra- and inter-hospital strain heterogeneity. However, the finding of isolates with indistinguishable pulsed-field types within the same ward and in two medical centers suggests patient-to-patient transmission or a common source. Four E. faecium strains were isolated within 48 h after admission of patients. CONCLUSIONS: These results indicate that VRE form part of the normal flora of patients and that, despite the actual scarcity of infections due to VRE, there is a potential risk for dissemination of these strains in French hospitals.

Journal Article↗
PubMed2004

Glycopeptide-resistance transferability from vancomycin-resistant enterococci of human and animal source to Listeria spp.

AIMS: The glycopeptide-resistance transferability from vancomycin-resistant enterococci (VRE) of clinical and animal origin to different species of Listeria was investigated. METHODS AND RESULTS: Of 36 matings, performed on membrane filter, the glycopeptide resistance was successfully transferred in six attempts, five with donors of animal origin and only one with donors from clinical source. The acquired glycopeptide resistance in Listeria transconjugants was confirmed by the presence of the conjugative plasmid band and by the amplification of the 732-bp fragment of vanA gene in transferred plasmids. CONCLUSIONS: Despite the lower number of bacteria used in this study, the source of enterococci influenced the outcome of mating. Moreover transferred VanA plasmid induced a different expression in Listeria transconjugants, suggesting that gene expression might be influenced by species affiliation of recipients. SIGNIFICANCE AND IMPACT OF THE STUDY: Our data strengthen the opinion that enterococci are an important source of resistance genes for Listeria via the transfer of movable genetic elements. As these strains are commonly found in the same habitats, a horizontal spread of glycopeptide resistance in Listeria spp. could be possible.

Animals↗
PubMed1999

Nucleotide sequence of IS1542, an insertion sequence identified within VanA glycopeptide resistance elements of enterococci.

IS1542 is an insertion sequence-like element which was originally identified in the orf2-vanR intergenic region of VanA resistance elements of glycopeptide-resistant Enterococcus faecium from hospital patients and from non-human sources in the UK. The nucleotide sequence of IS1542 was determined. It showed 81% homology with IS256 and contained an open reading frame sufficient to encode a 390 amino acid peptide predicted to have 87.2% identity with the transposase of IS256. By PCR, IS1542 was detected in the genomes of 26 of 28 (93%) VanA enterococci isolated from poultry in the UK and Ireland, but in only two of 65 (3%) glycopeptide-sensitive or VanC enterococci from hospital patients in the UK and Brazil. IS1542 may be a useful marker for evolutionary and epidemiological studies of VanA glycopeptide resistance in enterococci.

Amino Acid Sequence↗
PubMed1991

[Clinical features of enterococcal septicemia and antimicrobial susceptibilities for clinical isolates of enterococci in Nagoya University Hospital].

Enterococci are important causative organisms of nosocomial infections as well as methicillin resistant Staphylococcus aureus and Pseudomonas aeruginosa. Enterococcal bacteremia has been increased in Nagoya University Hospital from April 1983 to March 1990. We analyzed the clinical backgrounds, portal of entry and mortality for the patients with Enterococcal septicemia. Thirty-four cases with enterococcal septicemia (24 Enterococcus faecalis, 10 Enterococcus faecium) were subjected in this study. Twenty-seven cases were monomicrobial septicemia and 7 cases were polymicrobial septicemia. As to the portal of entry, intra-abdominal infections in 21 cases, urinary tract infections in 4 cases, respiratory tract infections or chest tubes in 2 cases, decbitus infection in 1 case, intravascular catheter in 1 case, and unknown sources in 5 cases. Total mortality of the septicemia was 44.1%. E. faecium and Enterococcus avium were much more resistant to antimicrobial agents than E. faecalis. Enterococci with high-level resistance to gentamicin (minimal inhibitory concentration, greater than 2000 micrograms/ml) accounted for 35% of the 26 clinical isolates of enterococci.

Adolescent↗
PubMed1992

Isolation rates and pathogenicity of enterococci in obstetric and gynecological operations.

Isolation of enterococci in patients undergoing obstetric and gynecological operations was studied as well as reviewing the postoperative infection due to this organism during the period from 1985 to 1990. 1) In 126 cases undergoing abdominal total hysterectomy, vaginal specimens were obtained before and after (3rd day) the operation. The isolation rates increased after the operation (before 16.7%, after 35.7%). They increased not only in the group using PIPC, CEZ, CEPR, CMZ, and LMOX by drip infusion but also in the group without prophylactic use of antibiotics. On the other hand in the group using CP vaginal suppositories, the isolation rate decreased. However no statistical proof was obtained as to antibiotics especially in regard to cephem drugs as the reason for the increase. 2) Enterococci were isolated from the surgical field during abdominal total hysterectomy in only 2.0% (n = 88). 3) Isolation rates of enterococci inside the transvaginal drain following radical hysterectomy (n = 30) reached 86.7%. 4) E. faecalis was isolated in 20.0% of the cases with wound infection (n = 25). However isolated Enterococcus strains were not regarded to be the causative organism. 5) There was one case of postoperative enterococcal septicemia in treating stage Ib adenocarcinoma of the uterine cervix.

Anti-Bacterial Agents↗
PubMed1977

Resistance to six aminoglycosidic aminocyclitol antibiotics among enterococci: prevalence, evolution, and relationship to synergism with penicillin.

Two hundred and three recent clinical isolates of enterococci were tested for susceptibility to streptomycin, kanamycin, amikacin, gentamicin, sisomicin, and tobramycin. Depending upon the source of the isolate, 36 to 54% of the enterococci demonstrated high-level resistance (minimal inhibitory concentration, >2,000 mug/ml) to streptomycin, 16 to 49% to kanamycin, and 0 to 14% to amikacin. None of the strains was highly resistant to gentamicin, sisomicin, or tobramycin. A comparison with isolates of enterococci obtained in 1968 revealed that there has been a decrease in prevalence of high-level resistance among organisms isolated from wound cultures in 1976. However, no decrease in resistance to streptomycin or kanamycin was demonstrated among blood or urine isolates. Penicillin, combined with gentamicin, sisomicin, or tobramycin, was synergistic against all 10 strains of Streptococcus faecalis subjected to formal testing. For streptomycin and kanamycin, the presence or absence of synergism with penicillin correlated with the absence or presence of high-level aminoglycoside resistance. High-level resistance to amikacin was seen in only 1 of the 10 strains. Nonetheless, combinations of penicillin plus amikacin failed to produce synergistic killing against 6 of the 10 strains. Indeed, the combination was synergistic only against those four strains that were susceptible to high levels of kanamycin.

Aminoglycosides↗
PubMed1987

Failure to demonstrate a consistent in vitro bactericidal effect of trimethoprim-sulfamethoxazole against enterococci.

Controversy exists as to the in vitro and in vivo activities of trimethoprim-sulfamethoxazole (TMP-SMX) against enterococci. In this study, we investigated the in vitro activity of TMP-SMX in the type of Mueller-Hinton broth previously reported to give the lowest MICs and MBCs with enterococci. In all instances, MICs were less than or equal to 0.5 microgram/ml. The majority of tests showed MBCs of greater than 32 micrograms/ml, although there was some effect from varying the inoculum and the length of incubation after subculturing. Minor differences were noted when tests were repeated and between the results from microdilution and macrodilution tests and those obtained by the time-kill method. These results, as well as other reports, suggest that TMP-SMX should not be considered a reliable bactericidal agent against enterococci.

Drug Combinations↗
PubMed1991

In vitro activity of RP 59500, a semisynthetic injectable pristinamycin, against staphylococci, streptococci, and enterococci.

The in vitro activity of RP 59500, a semisynthetic pristinamycin, was compared with the activities of vancomycin, oxacillin, ampicillin, gentamicin, ciprofloxacin, and rifampin against five Staphylococcus species, five Streptococcus species, and four Enterococcus species. For staphylococci, MICs were 0.13 to 1 microgram/ml and the MICs for 90% of the strains tested (MIC90s) were 0.13 to 0.5 microgram/ml; there were no differences between oxacillin-susceptible and -resistant strains. For streptococci, MICs were 0.03 to 4 micrograms/ml and MIC90s were 0.25 to 2 micrograms/ml; viridans group streptococci were the least susceptible streptococci. For enterococci, MICs were 0.25 to 32 micrograms/ml and MIC90s were 2 to 4 micrograms/ml; Enterococcus faecalis was the least susceptible. Vancomycin was the only comparative drug with consistent activity against all species of gram-positive cocci. With RP 59500, raising the inoculum 100-fold, lowering the pH of cation-adjusted Mueller-Hinton broth to 5.5, or omitting cation supplementation had little effect on MICs, but 50% serum increased MICs 2 to 4 dilution steps. The differences between MBCs and MICs were greater for staphylococci and enterococci than for streptococci. Time-kill studies with 24 strains indicated that RP 59500 concentrations 2-, 4-, and 16-fold greater than the MICs usually killed bacteria of each species at similar rates; reductions in CFU per milliliter were less than those observed with oxacillin or vancomycin against staphylococci and less than those observed with ampicillin against enterococci. RP 59500 antagonized the bactericidal activities of oxacillin and gentamicin against Staphylococcus aureus ATCC 29213 and that of ampicillin against E. faecalis ATCC 29212. Against the latter, combination with gentamicin was indifferent. RP 59500 has a broad spectrum of in vitro activity against gram-positive cocci; combining it with other drugs is not advantageous.

Anti-Bacterial Agents↗
PubMed1995

Activities of the semisynthetic glycopeptide LY191145 against vancomycin-resistant enterococci and other gram-positive bacteria.

LY191145 is the prototype of a series of compounds with activities against vancomycin-resistant enterococci derived by modification of the glycopeptide antibiotic LY264826. LY191145 had MICs for vancomycin- and teicoplanin-resistant enterococci of < or = 4 micrograms/ml for 50% of isolates and < or = 16 micrograms/ml for 90% of isolates. Its MICs for vancomycin-resistant, teicoplanin-susceptible enterococci were 1 to 8 micrograms/ml. LY191145 retains the potent activities of its parent compound against staphylococci and streptococci. In vivo studies in a mouse infection model confirmed these activities. This compound indicates the potential of semisynthetic glycopeptides as agents against antibiotic-resistant gram-positive bacteria.

Animals↗
PubMed1973

Synergism of vancomycin-gentamicin and vancomycin-streptomycin against enterococci.

The in vitro activity of vancomycin and combinations of vancomycin-gentamicin and vancomycin-streptomycin against enterococci was investigated. The minimal inhibitory concentration of vancomycin for 99 of 100 enterococcal strains isolated clinically was 3.12 mug or less/ml. When cultures of eight strains were incubated with vancomycin, regardless of the inoculum size (10(6), 10(5), or 10(4)) and concentration of vancomycin (10 or 20 mug/ml), there was no significant reduction in the number of viable enterococci at 6, 24, and 48 h. Gentamicin and streptomycin in concentrations attainable clinically were not effective against enterococci. Vancomycin combined with gentamicin or streptomycin was tested against 41 enterococcal strains. With the combination of vancomycin at 10 mug/ml and gentamicin at 4 mug/ml or vancomycin at 5 mug/ml and gentamicin at 4 mug/ml, synergism was demonstrated against all 41 strains at 6 h. The combination of vancomycin at 10 mug/ml and streptomycin at 10 mug/ml was only synergistic against 25 of 41 strains at 6 h, and only 22 of 41 strains were affected synergistically at 6 h by vancomycin at 5 mug/ml with streptomycin at 10 mug/ml. With few exceptions, the enhanced killing was more pronounced at 24 and 48 h. The combination of vancomycin and gentamicin or vancomycin and streptomycin (where in vitro studies demonstrate synergism) may be a useful alternate therapy in enterococcal endocarditis.

Bacteria↗
PubMed1996

In vitro activity of LY333328, an investigational glycopeptide antibiotic, against enterococci and staphylococci.

The in vitro activities of LY333328 were compared with those of vancomycin, teicoplanin, and quinupristin-dalfopristin (Synercid) against 219 strains of enterococci and staphylococci, including vancomycin-resistant enterococci and methicillin-resistant Staphylococcus aureus. MICs and MBCs were determined by a microtiter dilution protocol. LY333328 demonstrated superior activity against vancomycin-resistant enterococci and was the only antibiotic which was bactericidal. Its potency was comparable or superior to those of other antibiotics tested against methicillin-resistant staphylococci.

Anti-Bacterial Agents↗
PubMed1996

Semisynthetic glycopeptide antibiotics derived from LY264826 active against vancomycin-resistant enterococci.

Certain derivatives of the glycopeptide antibiotic LY264826 with N-alkyl-linked substitutions on the epivancosamine sugar are active against glycopeptide-resistant enterococci. Six compounds representing our most active series were evaluated for activity against antibiotic-resistant, gram-positive pathogens. For Enterococcus faecium and E. faecalis resistant to both vancomycin and teicoplanin, the MICs of the six semisynthetic compounds for 90% of the strains tested were 1 to 4 micrograms/ml, compared with 2,048 micrograms/ml for vancomycin and 256 micrograms/ml for LY264826. For E. faecium and E. faecalis resistant to vancomycin but not teicoplanin, the MICs were 0.016 to 1 micrograms/ml, compared with 64 to 1,024 micrograms/ml for vancomycin. The compounds were highly active against vancomycin-susceptible enterococci and against E. gallinarum and E. casseliflavus and showed some activity against isolates of highly vancomycin-resistant leuconostocs and pediococci. The MICs for 90% of the strains of methicillin-resistant Staphylococcus aureus tested were typically 0.25 to 1 micrograms/ml, compared with 1 microgram/ml for vancomycin. Against methicillin-resistant S. epidermidis MICs ranged from 0.25 to 2 micrograms/ml, compared with 1 to 4 micrograms/ml for vancomycin and 4 to 16 micrograms/ml for teicoplanin. The spectrum of these new compounds included activity against teicoplanin-resistant, coagulase-negative staphylococci. The compounds exhibited exceptional potency against pathogenic streptococci, with MICs of < or = 0.008 microgram/ml against Streptococcus pneumoniae, including penicillin-resistant isolates. In in vivo studies with a mouse infection model, the median effective doses against a challenge by S. aureus, S. pneumoniae, or S. pyogenes were typically 4 to 20 times lower than those of vancomycin. Overall, these new glycopeptides, such as LY307599 and LY333328, show promise for use as agents against resistant enterococci, methicillin-resistant S. aureus, and penicillin-resistant pneumococci.

Animals↗
PubMed1999

Antimicrobial susceptibility patterns of enterococci causing infections in Europe. The European VRE Study Group.

In vitro susceptibilities of 4,208 enterococci (83% Enterococcus faecalis isolates, 13.6% Enterococcus faecium isolates, and 3.4% isolates of other species) from patients in 27 European countries towards 16 antibiotics were determined. High-level resistance to gentamicin varied by country (range, 1 to 49%; mean, 22.6% +/- 12. 3%) and per species (19.7% E. faecalis isolates, 13.6% E. faecium isolates, 3.4% by other species). Vancomycin resistance was detected in 0.06% E. faecalis, 3.8% E. faecium, and 19.1% isolates of other species. All enterococci were susceptible to LY 333328 and everninomicin, and 25% of E. faecalis isolates and 85% of other enterococci were susceptible to quinupristin-dalfopristin. The MIC of moxifloxacin and trovafloxacin for ciprofloxacin-susceptible E. faecalis at which 90% of the isolates were inhibited was 0.25 to 0.5 microg/ml.

Anti-Bacterial Agents↗