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PubMed1998

Heterogeneity of resistance elements in clinical isolates of enterococci with high-level gentamicin resistance.

High-level resistance (minimum inhibitory concentration, MIC > 1,000 micrograms/ml) to gentamicin (HLGR) in enterococci is common in Taiwan. In this study, we investigated the distribution of gentamicin resistance elements in enterococci isolated at National Taiwan University Hospital in a 1-year period, and also examined the transfer and the genetic variability of the resistance elements of different isolates. Among 109 isolates tested, 43 (39%) HLGR isolates were identified. HLGR was most common in Enterococcus faecium isolates (7/15, 47%), followed by Enterococcus faecalis (34/80, 43%), Enterococcus avium (1/5, 20%), and Enterococcus casseliflavus (1/9, 11%). To understand the mechanism of resistance transfer, four isolates of E. faecalis and five isolates of E. faecium showing HLGR were studied. Transfer of resistance markers to a plasmid-free recipient strain of E. faecalis JH2-7 was observed, with transfer frequencies ranging from 10(-2) to 10(-8). All of the transconjugants contained plasmids, with sizes ranging from 45 kb to larger than 70 kb. At least three plasmid patterns were observed on digestion with HaeIII. Hybridization with a probe specific for the aac6'aph2" gentamicin resistance gene confirmed that all of these HLGR isolates carried a Gm(r) determinant, though the hybridization patterns of the plasmids from E. faecalis and E. faecium were different. Although many similarities exist among enterococcal Gm(r) determinants, the results suggest heterogeneity may occur in the flanking regions of resistance elements.

DNA Transposable Elements↗

PubMed2003

Multicentre evaluation of the in vitro activity of linezolid in the Western Pacific.

Multiresistance to antimicrobial agents is common in staphylococci and pneumococci isolates in the Western Pacific region. The activity of linezolid, a new oxazolidinone, was evaluated against a spectrum of Gram-positive species collected in the region. Eighteen laboratories from six countries in the Western Pacific examined the linezolid susceptibility of 2143 clinical isolates of Staphylococcus aureus, coagulase-negative staphylococci (CoNS) and Enterococcus spp. using broth microdilution or disc diffusion methodology. For Streptococcus pneumoniae (n = 351) and other streptococci (n = 83), Etest (AB Biodisk, Solna, Sweden) strips were used. Results were compared with other common and important antimicrobials. Linezolid-resistant strains were not detected among streptococci or staphylococci, including a significant proportion of S. aureus strains that were multiresistant. Almost all enterococci, including 14 vancomycin-resistant Enterococcus faecium, were linezolid susceptible. A small proportion of enterococci (0.8%) were intermediate to linezolid, and one strain of Enterococcus faecalis had a zone diameter of 20 mm (resistant). The linezolid MIC ranges (MIC(90)) of those strains tested by broth microdilution or Etest were: 1-4 mg/L (2 mg/L) for S. aureus, 0.5-4 mg/L (2 mg/L) for CoNS, 0.5-4 mg/L (2 mg/L) for Enterococcus spp., 0.12-2 mg/L (1 mg/L) for S. pneumoniae and 0.25-2 mg/L (1 mg/L) for Streptococcus spp. There was no difference in linezolid susceptibility between countries or between multiresistant and susceptible strains of each species monitored.

Acetamides↗

PubMed1995

Investigation of the reformulated Remel Synergy Quad plate for detection of high-level aminoglycoside and vancomycin resistance among enterococci.

We investigated the accuracy of the recently released Remel Synergy Quad plate, a commercially available agar screening method for detecting high-level aminoglycoside and vancomycin resistance among enterococci that is based on the National Committee for Clinical Laboratory Standards recommended guidelines (National Committee for Clinical Laboratory Standards, M7-A3, 1993). The Synergy Quad correctly determined the gentamicin and streptomycin resistance status for > or = 97% of 147 Enterococcus faecalis and Enterococcus faecium isolates tested. Detection of vancomycin resistance also was reliable, as no false susceptibility occurred with 36 vancomycin-resistant E. faecalis and E. faecium strains and false resistance occurred only once with the 47 susceptible strains tested. One strain each of Enterococcus gallinarum and Enterococcus casseliflavus failed to grow on the screen, but because the true nature and significance of resistance in such isolates is unknown the implication of their screen negativity is uncertain. In summary, the Remel Synergy Quad provides a highly accurate and convenient method for susceptibility testing of enterococci against gentamicin, streptomycin, and vancomycin.

Aminoglycosides↗

PubMed2004

Adhesion properties of enterococci to intestinal mucus of different hosts.

The adhesive capacity of selected enterococci to human, canine and porcine intestinal mucus was investigated in order to select for potential probiotic strains with good adhesive properties for human or animal use. The adhesion to the human intestinal mucus of the tested strains was found to range from log10 3.8 to log10 8.6 cfu per microtitre plate well. The highest adhesion to the human intestinal mucus was found among strains from horse faeces, dog faeces and dog feed. The adhesion to canine mucus was observed to range from log10 3.8 to log10 8.3 cfu/well, with the highest adhesive capacity among strains from dog faeces, horse faeces and dog's feed; on average log10 7.9, 7.3 and 7.0 cfu/well, respectively. Isolates from dogs did not bind at higher levels to canine mucus than to human mucus. A strong correlation was observed for the adhesion to human and canine intestinal mucus (p < 0.0001) and also between porcine and canine or human mucus (p < 0.05 for both). When comparing the adhesion of Enterococcus faecium and E. faecalis, no statistical significant differences were observed for any of the tested mucus types. The tested Enterococcus strains were found to exhibit a strain dependent on in vitro adhesion to human, canine and porcine intestinal mucus and did not exhibit host specificity in their adhesion, though some sources appeared to contain more adhesive strains than others. To our knowledge, this is the first report on the in vivo adhesion to intestinal mucus of a large number of enterococci from different sources.

Animal Feed↗

PubMed2003

Vancomycin-resistant Enterococcus faecalis colonization during recovery from Neisseria meningitidis cerebrospinal meningitis. Case report.

A 19-year-old man had been admitted to the Hospital because of septic shock and large scale suffusions all over the body. The pathogen had proved to be Neisseria meningitidis serogroup C. In his stabilization period two superinfectious attacks arose. One of them was a bacteremia, caused by a vancomycin-sensitive Enterococcus faecium. The second was a wound infection in his deep colliquating necrotised tissue of the heel. Vancomycin-resistant Enterococcus faecalis (VREF) was isolated from this lesion with some Gram-negative opportunistic pathogens. The strain contained the vanA gene. After systemic and topical treatment, furthermore plastic surgical interventions the patient recovered. This is the second report on VREF from Hungary colonizing/infecting a patient with an underlying disease.

Adult↗

PubMed1999

Proficiency of clinical laboratories in Spain in detecting vancomycin-resistant Enterococcus spp. The Spanish VRE Study Group.

Studies in a variety of U.S. clinical laboratories have demonstrated difficulty in detecting intermediate and low-level vancomycin-resistant enterococci (VRE). The misclassification of "at least intermediate resistant isolates" as vancomycin susceptible may have both clinical implications and a negative impact on measures to control the spread of VRE. No published study has assessed the ability of clinical laboratories in Europe to detect VRE. So, the apparent low prevalence of VRE in European hospitals may be, in part, secondary to the inability of these laboratories to detect all VRE. In an effort to assess European laboratories' proficiency in detecting VRE, we identified 22 laboratories in Spain and asked them to test four VRE strains and one susceptible enterococcal strain from the Centers for Disease Control and Prevention collection. Each organism was tested by the routine antimicrobial susceptibility testing method used by each laboratory. Overall, VRE were correctly identified in 61 of 88 (69.1%) instances. The accuracy of VRE detection varied with the level of resistance and the antimicrobial susceptibility method. The high-level-resistant strain (Enterococcus faecium; MIC, 512 microg/ml) was accurately detected in 20 of 22 (91. 3%) instances, whereas the intermediate-resistant isolate (Enterococcus gallinarum; MIC, 8 microg/ml) was accurately detected in only 11 of 22 (50%) instances. Classification errors occurred in 27 of 88 (30.9%) instances. Misclassification as vancomycin susceptible was the most common error (16 of 27 [59.3%] instances). Our study shows that the participating Spanish laboratories had an overall acceptable proficiency in detecting VRE but that a substantial proportion of VRE isolates with low or intermediate levels of resistance were not detected. We recommend that studies be conducted to validate laboratory proficiency testing as an important step in the prevention and control of the spread of antimicrobial resistance.

Centers for Disease Control and Prevention, U.S.↗

PubMed2001

Antimicrobial susceptibility patterns of enterococci in intensive care units in Sweden evaluated by different MIC breakpoint systems.

Three hundred and twenty-two (322) clinical isolates were collected from patients admitted to intensive care units (ICUs) at eight Swedish hospitals between December 1996 and December 1998. Of the isolates, 244 (76%) were Enterococcus faecalis, 74 (23%) were Enterococcus faecium and four (1%) were other Enterococcus spp. MICs of ampicillin, imipenem, meropenem, piperacillin/tazobactam, ciprofloxacin, trovafloxacin, clinafloxacin, gentamicin, streptomycin, vancomycin, teicoplanin, quinupristin/dalfopristin, linezolid and evernimicin were determined by Etest. Susceptible and resistant isolates were defined according to the species-related MIC breakpoints of the British Society for Antimicrobial Chemotherapy (BSAC), the National Committee for Clinical Laboratory Standards (NCCLS) and the Swedish Reference Group for Antibiotics (SRGA). Tentative breakpoints were applied for new/experimental antibiotics. Multidrug resistance among enterococci in ICUs is not uncommon in Sweden, particularly among E. faecium, and includes ampicillin resistance and concomitant resistance to fluoroquinolones. Almost 20% of E. faecalis isolates showed high-level resistance to gentamicin and concomitant resistance to fluoroquinolones. Vancomycin-resistant enterococci were only found sporadically. Among the new antimicrobial agents, linezolid and evernimicin showed the best activity against all enterococcal isolates. There was good concordance between the BSAC, NCCLS and SRGA breakpoints in detecting resistance. When applying the SRGA breakpoints for susceptibility, isolates were more frequently interpreted as intermediate. This might indicate earlier detection of emerging resistance using the SRGA breakpoint when the native population is considered susceptible, but with the risk that isolates belonging to the native susceptible population will be incorrectly interpreted as intermediate.

Anti-Bacterial Agents↗

PubMed1999

Probiotics shown to change bacterial community structure in the avian gastrointestinal tract.

Culturing and molecular techniques were used to monitor changes in the bacterial flora of the avian gastrointestinal (GI) tract following introduction of genetically modified (GM) and unmodified probiotics. Community hybridization of amplified 16S ribosomal DNA demonstrated that the bacterial flora of the GI tract changed significantly in response to the probiotic treatments. The changes were not detected by culturing. Although both GM and non-GM strains of Enterococcus faecium NCIMB 11508 changed the bacterial flora of the chicken GI tract, they did so differently. Probing the community DNA with an Enterococcus faecalis-specific probe showed that the relative amount of E. faecalis in the total eubacterial population increased in the presence of the non-GM strain and decreased in the presence of the GM probiotic compared with the results obtained with an untreated control group.

Aging↗

PubMed2000

Comparison of different reducing agents for enhanced detection of heat-injured Listeria monocytogenes.

The effect of different reducing agents (L-cysteine, Oxyrase, and Enterococcus faecium) and their combinations on the detection of heat-injured (62.8 degrees C, 7.5 min or 10 min) Listeria monocytogenes was examined. The incorporation of L-Cysteine (0.5 g/liter) yielded higher percentage detection than any of the other reducing agents (P < 0.05). The combination of Oxyrase (10 U/ml) and E. faecium (10(3) CFU/ml) synergistically enhanced the detection of L. monocytogenes heat-injured for 10 min at 62.8 degrees C (P < 0.05). Simultaneous addition of L-cysteine (0.5 g/liter) and Oxyrase (10 U/ml) significantly lowered the recovery of heat-injured L. monocytogenes (P < 0.05). Higher activities of Oxyrase (50 U/ml) inhibited the detection of heat-injured L. monocytogenes (P < 0.05). The rates of depletion of relative percentage O2 were in the order: L-cysteine (0.5 g/liter; 6.63%/ min) > Oxyrase (10 U/ml; 5.00%/min) > E. faecium (10(8) CFU/ml; 1.66%/min) > E. faecium (10(3) CFU/ml; 0.20%/min). The final levels of redox potential (Eh) achieved were -110.5 mV, -100 mV, -83.5 mV, and -25 mV for E. faecium (10(8) CFU/ml), L-cysteine, Oxyrase, and E. faecium (10(3) CFU/ml), respectively.

Animals↗

PubMed2002

Antimicrobial activities of naphthazarins from Arnebia euchroma.

Bioassay-directed fractionation of extract of Arnebia euchroma led to the isolation of alkannin (1), shikonin (2), and their derivatives (3-8) as the active principles against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE). The stereochemistry of alpha-methylbutyryl alkannin (8) is revealed for the first time, and the antimicrobial activity of 8 was compared with its corresponding diastereomer (9). The derivatives 3-9 showed stronger anti-MRSA activity [minimum inhibitory concentrations (MICs) ranged from 1.56 to 3.13 microg/mL] than alkannin or shikonin (MIC = 6.25 microg/mL). Anti-MRSA activity of derivatives was bactericidal with minimum bactericidal concentration (MBC)/MIC < or = 2. In a time-kill assay, the bactericidal activity against MRSA was achieved as rapidly as 2 h. The derivatives 3-9 were also active against vancomycin-resistant Enterococcus faecium (F935) and vancomycin-resistant Enterococcus faecalis (CKU-17) with MICs similar to those with MRSA. Aromatic ester derivatives were also synthesized for antimicrobial activity comparison. None of these compounds were active against Gram-negative bacteria tested. Their cytotoxicity was also evaluated on selected cancer cell lines, and they expressed their activity in the range 0.6-5.4 microg/mL (CD(50)). Our results indicate that the ester derivatives of alkannin are potential candidates of anti-MRSA and anti-VRE agents with antitumor activity.

Anti-Bacterial Agents↗

PubMed2003

Pharmacodynamic profile of daptomycin against Enterococcus species and methicillin-resistant Staphylococcus aureus in a murine thigh infection model.

OBJECTIVE: To describe the pharmacodynamic profile of daptomycin against methicillin-resistant Staphylococcus aureus (MRSA) and Enterococci species based on bacterial density in an immunocompromised mouse thigh infection model. MATERIALS AND METHODS: The pharmacodynamic (PD) profile of daptomycin was determined against two MRSA, one vancomycin-resistant Enterococcus faecium, and one vancomycin-susceptible Enterococcus faecalis using the immunocompromised murine thigh model. Efficacy was assessed by the change in log10 cfu in thighs after 24 h of drug treatment. RESULTS: Daptomycin produced a maximal kill of 4.5-5 log10 cfu against the MRSA and 1.5-2 log10 for the Enterococcus species. AUC/MIC was the most predictive of the PD parameters. Utilizing MICs determined in serum or broth in the calculation of the PD parameters had minimal effect on this correlation. AUCfree/MICbroth required for static effects with MRSA and Enterococcus species were 12-36 and 5-13, whereas 99% of maximal kill was achieved at ratios of 171-442 and 38-157, respectively. CONCLUSIONS: These data reveal the potent in vivo bactericidal activity of daptomycin against MRSA and Enterococcus species using clinically achievable drug exposures (dose 4-6 mg/kg per day) currently under investigation in man.

Animals↗

PubMed2002

In vitro assessment of urinary isolates of ampicillin-resistant enterococci.

OBJECTIVE: Susceptibility and minimum inhibitory concentration (MIC) studies of ampicillin-resistant enterococci (ARE) were performed with vancomycin, ciprofloxacin, and trovafloxacin. Ampicillin MICs were determined to make comparisons with achievable urinary concentrations of ampicillin. DESIGN: From July 1998 to April 1999, all enterococci isolated from urinary specimens were tested for susceptibility to ampicillin by disk diffusion. For all ARE, vancomycin, ciprofloxacin, and trovafloxacin susceptibilities were determined by use of either disk diffusion or the E-test. Ampicillin MICs were determined for these isolates by liquid agar microdilution testing. ARE were identified to the species level on the basis of biochemical reactions. SETTING: The study was performed at a university-affiliated tertiary care hospital. OUTCOME MEASURES: In vitro susceptibility studies and MIC determinations were performed in accordance with the National Committee for Clinical Laboratory Standards. RESULTS: A total of 310 urine samples were culture positive for enterococcus. Thirty (9.7%) unduplicated isolates were resistant to ampicillin. Of these, nine ARE isolates (30%) were also vancomycin resistant, whereas only 2 ampicillin-susceptible isolates were vancomycin resistant (p < 0.05). All ARE were resistant to ciprofloxacin, and 29 (96.7%) were resistant to trovafloxacin. Nine (30%), 18 (60%), and 3 (10%) isolates had an ampicillin MIC of 128, 256, and 512 micrograms/mL, respectively. Ampicillin MICs did not differ significantly between vancomycin-susceptible and -resistant isolates (p = 0.963). Twenty-seven isolates (90%) were identified as Enterococcus faecium; the other 3 were either Enterococcus avium or Enterococcus raffinosus. CONCLUSIONS: Ampicillin resistance is associated with resistance to vancomycin. Most ARE are resistant to fluoroquinolone antibiotics such as ciprofloxacin and trovafloxacin. Ampicillin MICs for ARE found in these urinary specimens were all within 1 dilution of 256 micrograms/mL, a concentration achievable in the urine with higher doses of oral amoxicillin or intravenous ampicillin. Additional studies are needed to assess the clinical implications of these data.

Adult↗

PubMed2000

Quinupristin-dalfopristin: an overview.

Synercid (RP 59500), the first injectable streptogramin antibiotic, is composed of two semisynthetic pristinamycin derivatives, quinupristin and dalfopristin. Individually, each component has bacteriostatic activity against staphylococci and streptococci, but together, the agents exhibit synergy, leading to bactericidal activity. The combination drug, however, is bacteriostatic against Enterococcus faecium and has poor activity against Enterococcus faecalis. Despite a short half-life, an extended postantibiotic effect allows the agent to be dosed every 8-12 hours. Both drugs are largely hepatically metabolized and excreted in bile. Although not metabolized by cytochrome P450 3A4, quinupristin-dalfopristin can inhibit agents that are metabolized through this pathway. Dosage adjustments may be necessary in patients with hepatic dysfunction. Alterations in renal function have minimal effects on the agent's pharmacokinetics. Adverse events include arthralgia, myalgias, and infusion-related pain. Based on available data, quinupristin-dalfopristin appears to have a role in treating severely ill patients with infections due to multiresistant gram-positive pathogens.

Animals↗

PubMed1999

Evaluation of the revised MicroScan dried overnight gram-positive identification panel to identify Enterococcus species.

The revised MicroScan Dried Overnight Gram-Positive Identification panel was evaluated for its efficacy at identifying Enterococcus species in comparison with conventional biochemical tests. Supplemental testing of ampicillin-susceptible Enterococcus faecium for motility and the ability to acidify methyl-alpha-D-glucopyranoside helped recognize E. gallinarum and increased the accuracy of the panel for identifying Enterococcus species to 98.5%.

Ampicillin Resistance↗

PubMed2002

Microbicidal efficacy of superheated steam. II. Studies involving E. faecium and spores of B. xerothermodurans and B. coagulans.

The authors' studies on the heat resistance of microorganisms and the superheating of steam (Spicher et al., Zbl. Hyg. Umweltmed. 201, 541-553, 1999) have been continued, using the vegetative bacterium, Enterococcus faecium, and spores of B. xerothermodurans and B. coagulans. The temperature of the saturated steam was 68 degrees C (E. faecium), 105 degrees C (B. xerothermodurans) and 110 degrees C (B. coagulans), respectively. The steam was superheated by 30-40 K, as a maximum. The test organisms had been fixed to fibre glass fleece. The time of exposure to saturated or superheated steam after which 50% of the bioindicators yielded no viable germs was used as a measure of resistance of the organisms. In the discussion, reference has also been made to the findings for B. subtilis and B. stearothermophilus spores obtained in the preceding study. E. faecium exhibited a maximum resistance when superheating the steam by 5 K. The superheated steam required a 74-fold exposure time compared to saturated steam of 68 degrees C. Resistance became gradually reduced as superheating was further increased. Even superheating by 30 K required a 11-fold exposure time. The bacterial spores exhibited maxima of resistance on superheating by 23-30 K. In these cases necessary exposure times were 119 (B. subtilis), 30 (B. xerothermodurans), and 4 times (B. coagulans, B. stearothermophilus) longer than those required in saturated steam. In the superheating range below 10 K, behaviour patterns varied. Thus, heat resistance may initially become reduced with increasing superheating (B. coagulans), remain on almost the same level as under exposure to saturated steam (B. stearothermophilus), reach a stage of weakly enhanced resistance (B. xerothermodurans), or approach a maximum of resistance in an almost linear mode (B. subtilis). It appears that there are differences between strains of one and the same bacterial species.

Adaptation, Physiological↗

PubMed1999

Comparison of synthesis and antibacterial activity of temporin A.

Temporin A is a small, basic, highly hydrophobic, antibacterial peptide found in the skin of the European red frog, Rana temporaria. It was synthesized twice by the FastMoc solid phase method using amino acids protected at the N(alpha)-position with either 9-fluorenylmethoxycarbonyl or 2-(4-nitrophenylsulfonyl)ethoxycarbonyl. The syntheses of temporin A demonstrates the difference between 2-(4-nitrophenylsulfonyl)ethoxycarbonyl and 9-fluorenylmethoxycarbonyl amino acids. The purified peptide showed also antibacterial activity against clinically important gram-positive bacteria. It was found to have a moderately good activity against both methicillin resistant and sensitive strains of Staphylococcus aureus, but a weaker activity against vancomycin resistant strains of Enterococcus faecium.

Animals↗

PubMed1998

Susceptibility patterns of Enterococcus spp. isolated in Poland during 1996.

Susceptibility of Enterococcus spp. isolated from various clinical specimens to different antimicrobial agents was evaluated. Of the 346 enterococcal isolates obtained from four regional Polish hospitals during 6 months of 1996, 261 (75.4%) were identified as Enterococcus faecalis, 75 (21.7%) as Enterococcus faecium and ten (2.9%) as other enterococcal species. High-level resistance to gentamicin was expressed by 33.4% of E. faecalis and 86.5% of E. faecium strains and corresponding streptomycin resistance by 43.9 and 82.4%, respectively. Over 80% of E. faecium isolates were resistant to ampicillin. None of the isolates was resistant to teicoplanin, however 7.9% of E. fecalis and 1.4% of E. faecium strains were moderately susceptible to vancomycin.

Anti-Bacterial Agents↗

PubMed2004

[Susceptibility of Enterococcus genus to new antimicrobial agents].

The aim of this study was to determine the prevalence of resistance to different antibiotics in 109 clinical strains of Enterococcus isolated consecutively in hospital over a four-month period in 2002. The strains were identified by species using the semiautomatic system Microscan walk away 40 (Dade Behring) and the API20 STREP system (Biomerieux). Three different methods for the susceptibility study were used: Sensititre, E-test and disc diffusion. The percentage of Enterococcus faecalis and Enterococcus faecium isolated was 92% and 8%, respectively. The following resistance to Enterococcus faecalis was detected: erythromycin (53%), telithromycin (36%), penicillin (2%), ampicillin (1%), vancomycin, teicoplanin and linezolid (0%), high level resistance to streptomycin (51%) and gentamicin (32%). A high percentage of strains resistant to penicillin and ampicillin was detected in E. faecium (six of nine strains). For two species, high susceptibility to linezolid and glycopeptides was found.

Drug Resistance, Bacterial↗