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PubMed1995

Resistance to aminoglycosides and other antibiotics among clinical isolates of Enterococcus spp.

We conducted a study on 81 clinical isolates of enterococci (65 Enterococcus faecalis and 16 Enterococcus faecium) collected consecutively over a one year period at our University Hospital. Of them, 10 (15%) Enterococcus faecalis and 6 (38%) Enterococcus faecium showed high level resistance to aminoglycosides, were all acquired in surgical units and in five cases they were responsible for invasive infections. Ampicillin and imipenem were active only against Enterococcus faecalis. Vancomycin and teicoplanin were bacteriostatic; however, teicoplanin MICs were lower and in 45% of cases its MBC values were within clinically achievable serum concentrations. Ciprofloxacin and cotrimoxazole were active in vitro.

Aminoglycosides↗

PubMed2000

Epidemiological, microbiological, clinical, and prognostic factors of bacteremia caused by high-level vancomycin-resistant Enterococcus species.

A case-control study was performed between 1994 and 1996 in order to study the epidemiological, microbiological, clinical, and prognostic features of high-level vancomycin-resistant enterococcal bacteremia. Seventeen consecutive patients who had clinically significant bacteremia due to vancomycin-resistant enterococci (vanA genotype: 16 Enterococcus faecalis, 1 Enterococcus faecium) were compared with 169 who had vancomycin-susceptible enterococcal bacteremia. The following were selected by multivariate analysis as independent risk factors that influenced the development of high-level vancomycin-resistant enterococcal bacteremia: prior glycopeptide therapy (P=0.049); inclusion in a hemodialysis program (P=0.046); prior therapy with corticosteroids or antineoplastic agents (P=0.029); and prior surgical treatment (P=0.022). The following other factors were selected by univariate analysis: tracheostomy (P=0.002); prolonged hospitalization (P=0.01); and any kind of puncture (P=0.02). The crude associated-mortality rate was 13.4%. Gene amplification of vanA was positive for 17 strains of enterococci. Pulsed-field gel electrophoresis of genomic DNA after SmaI digestion of vanA isolates revealed that one strain predominated (10 isolates), though at least four similar banding patterns were identified (6 isolates). The 16 strains closely related to the outbreak were investigated further. The surgical intensive care unit was the first and most involved service. The hospital outbreak of vanA vancomycin-resistant enterococcal bacteremia occurred between 1994 and 1995 and was caused by Enterococcus faecalis. This is believed to be the first and only such outbreak described in a Spanish hospital thus far.

Adolescent↗

PubMed2005

Antimicrobial resistance of enterococci in Turkey.

Susceptibility to ampicillin, penicillin, vancomycin and teicoplanin, high-level resistance to aminoglycosides (gentamicin and streptomycin) and beta-lactamase production were investigated among 264 consecutive clinical enterococcal isolates in Turkey. Disc diffusion test was used to detect resistance to ampicillin, penicillin, vancomycin and teicoplanin. High-level resistance to aminoglycosides was determined both by standard agar screening and by disc diffusion methods. The values of minimum inhibitory concentration (MIC) of each isolate for ampicillin, vancomycin and teicoplanin were determined by the microbroth dilution technique. The isolates were found to consist of Enterococcus faecalis (78%), Enterococcus faecium (9%) and Enterococcus spp. (12%). In all strains, the penicillin and ampicillin resistance ratios were 27% and 26%, respectively. Enterococcus faecalis was more susceptible to penicillin and ampicillin than the other strains. None of the strains were resistant to glycopeptides. High-level aminoglycoside resistance was found in 16% E. faecalis and 88% E. faecium for gentamicin, and 35% and 44%, respectively, for streptomycin. There were no differences between the two methods used to determine the aminoglycoside resistance rates in the enterococcal isolates. No beta-lactamase-producing isolates were detected in either species. In conclusion, to determine the resistance of enterococci to the penicillin group of drugs by the disc diffusion method, both penicillin and ampicillin discs should be evaluated. In serious enterococcal infections, before starting combined therapy, high-level aminoglycoside resistance should be investigated.

Aminoglycosides↗

PubMed1996

Comparative in vitro activity of levofloxacin and ofloxacin against gram-positive bacteria.

The in vitro activity of levofloxacin against 506 Gram-positive bacteria was compared with those of D(-)-ofloxacin, ofloxacin, ciprofloxacin, and sparfloxacin. Levofloxacin was generally twice as active as ofloxacin against these organisms (range, 0-3 twofold dilutions). Sparfloxacin appeared to have the greatest activity overall, but for several groups of organisms minimum inhibitory concentrations (MIC90s) of this compound were within one twofold dilution of those of levofloxacin. Resistance to levofloxacin (MIC > or = 8 micrograms/ml) was not encountered among streptococcal species, was rare in methicillin-susceptible staphylococci (1.7%), and was infrequent in vancomycin-susceptible Enterococcus faecalis and Enterococcus faecium (8.7%). Resistance was more common among vancomycin-resistant enterococci and methicillin-resistant staphylococci.

Anti-Bacterial Agents↗

PubMed2006

Antimicrobial resistance and the mechanisms implicated in faecal enterococci from healthy humans, poultry and pets in Portugal.

Antimicrobial resistance and the mechanisms implicated were studied in 440 enterococci (227 Enterococcus faecium, 177 Enterococcus faecalis, 32 Enterococcus hirae and 4 Enterococcus durans) recovered from 220 faecal samples of healthy humans, poultry and pets in Portugal. Higher levels of resistance were detected for ampicillin, tetracycline, erythromycin and chloramphenicol in poultry isolates (10.5%, 97%, 87.5% and 16%, respectively) compared with human isolates (0%, 26%, 31.5% and 5%, respectively); intermediate levels of resistance for these antibiotics were found in pet isolates. Thirty-three per cent of the E. faecium isolates of poultry origin showed quinupristin/dalfopristin resistance. High-level resistance to gentamicin or streptomycin was detected in 1-7% of isolates in our series of enterococci. The aac(6')-aph(2''), aph(3')-IIIa, erm(B) and tet(M) genes were demonstrated in most of the gentamicin-, kanamycin-, erythromycin- and tetracycline-resistant isolates, respectively. The vat(E) gene was found in 39% of the quinupristin/dalfopristin-resistant E. faecium isolates of poultry origin.

Animals↗

PubMed1999

Surveillance of antibiotic resistance in ICUs in southeastern Sweden. ICU Study Group of the South East of Sweden.

BACKGROUND: A study was designed to assess a computer-based program for continuous registration of antibiotic resistance, statistics concerning severity of illness, and consumption of antibacterial drugs. METHODS: The frequency of antibiotic resistance among bacteria in eight ICUs in southeastern Sweden was investigated yearly from 1995 through 1997. The antibiotic consumption in the ICUs was registered as defined daily doses (DDD) and compared to severity of illness (APACHE-II scores). RESULTS: There was a statistically significant increase in ampicillin resistance among Enterococcus spp. between 1996 and 1997, which was due to a shift from Enterococcus faecalis to Enterococcus faecium. A high prevalence of resistance among coagulase-negative staphylococci to oxacillin (approximately 70%), ciprofloxacin (approximately 50%), fucidic acid (approximately 50%) and netilmicin (approximately 30%) was seen in all ICUs during the whole study period. There was a statistically significant increase in ciprofloxacin resistance among Escherichia coli and Enterococcus spp. The resistance among Enterobacter spp. to cefotaxime decreased but this change was not statistically significant. Efforts were made to avoid betalactam antibiotics, except carbapenems, for treatment of infections caused by Enterobacter spp. and the consumption of cephalosporins decreased whereas the consumption of carbapenems increased. The total antibiotic consumption decreased by 2.5% during the study period. There was no correlation between APACHE II scores and antibiotic consumption. CONCLUSIONS: Each ICU within a hospital ought to have a program for "on-line" antibiotic resistance surveillance of drugs used in that unit so that changes in empirical treatment can be made when there is an increase in antibiotic-resistant isolates within that unit.

APACHE↗

PubMed2003

Species prevalence and antibacterial resistance of enterococci isolated in Kuwait hospitals.

This study investigated the species prevalence and antibacterial resistance among enterococci isolated in Kuwait hospitals. They consisted of 415 isolates of Enterococcus faecalis (85.3 %), Enterococcus faecium (7.7 %), Enterococcus casseliflavus (4.0 %), Enterococcus avium (1.2 %), Enterococcus durans (1.0 %), Enterococcus gallinarium (0.5 %) and Enterococcus bovis (0.2 %) isolated from urine (36.6 %), blood (10.4 %), wound swabs (11.0 %), stool samples (12.0 %), high vaginal swabs (9.0 %), endocervical swabs (3.0 %) and miscellaneous sources (18.0 %). All of them were susceptible to linezolid. Fifty-two (12.5 %) isolates were ampicillin resistant but none of them produced beta-lactamase. They were resistant to erythromycin (63.3 %), tetracycline (60.5 %), ciprofloxacin (40.0 %), chloramphenicol (28.0 %), vancomycin (2.6 %), and teicoplanin (2.6 %). Fourteen, 19 and 20 % of them expressed high-level resistance to gentamicin, kanamycin and streptomycin, respectively. All of the vancomycin-resistant strains carried the vanA phenotype and genotype. There was no evidence of clonal spread of the vancomycin-resistant isolates.

Acetamides↗

PubMed2004

[Vancomycin resistant enterococci in the Netherlands].

Enterococci (Enterococcus faecalis and Enterococcus faecium) are relatively avirulent enteric bacteria that usually only cause infections in immunocompromised patients. Antimicrobial treatment, however, is hampered as enterococci are intrinsically resistant to many antibiotics. For years, vancomycin was considered the last available antibiotic. Plasmid-mediated resistance against vancomycin among enterococci was first described in the nineteen-eighties and since then incidences of infection caused by vancomycin-resistant enterococci (VRE) have increased dramatically, especially in the United States. In 2000, three outbreaks of VRE occurred in hospitals in the Netherlands and a set of infection-control measures was proposed to limit further transmission. These measures were based on the simultaneous isolation of VRE from multiple patients. All three outbreaks were controlled by these measures and no new outbreaks in Dutch hospitals have been reported since then. Epidemiological studies have shown that hospital outbreaks on three continents were caused by a subpopulation of E. faecium, which is characterized by the presence of a potential virulence gene (variant esp) and resistance to amoxicillin. This 'hospital strain' of E. faecium has probably been prevalent within hospital settings for some time, but only became clinically relevant when it had acquired vancomycin-resistance. Current advice is to implement the set of infection control measures formulated in 2000, only in those patients colonized by amoxicillin-resistant VRE. The potential dangers of VRE were recently underlined by the proven transmission of the vancomycin-resistance gene from VRE to methicillin-resistant Staphylococcus aureus (MRSA) in two patients in the United States. It is in the interest of the patients that prevalence of VRE and MRSA in Dutch hospitals should be kept as low as possible.

Disease Outbreaks↗

PubMed1992

[Studies of bacterial indicators for water pollution--growth of Escherichia coli and enterococci in limited nutrient conditions].

Bacteria, that would be an ideal indicator for fecal contamination in environmental water, should not proliferate but at the same time should survive a little longer than enteropathogenic bacteria in the environmental water. While conventional methods have recommended Escherichia coli as an indicator, enterococci is recommended as a preferable indicator for the estimation of fecal contamination based on the following observation. The relationship between the specific growth rate of organism and the concentration of peptone was determined and the value of Ks, a parameter analogous to Michaelis-Menten constant, was estimated. Ks of Escherichia coli, Enterococcus faecalis and Enterococcus faecium were 3.8, 11.1 and 12.5 mg/ml respectively, indicating that Escherichia coli will grow at lower concentration of peptone than enterococci. Escherichia coli also grew in human fecal suspension and in filtrated (0.22 microns) river water, and while in those conditions enterococci did not grow or survive longer than Escherichia coli, survival time was longer than for Salmonella typhi and Shigella sonnei. It appears that enterococci reflect more accurately actual fecal contamination.

Enterococcus faecalis↗

PubMed1996

In vitro conjugative transfer of VanA vancomycin resistance between Enterococci and Listeriae of different species.

In a study designed to gain data on the in vitro transferability of vancomycin resistance from enterococci of the VanA phenotype to listeriae of different species, three clinical Enterococcus isolates-Enterococcus faecium LS10, Enterococcus faecalis LS4, and Enterococcus faecalis A3208, all harboring a plasmid that strongly hybridized with a vanA probe-were used as donors in transfer experiments. Strains of five Listeria species were used as recipients. From Enterococcus faecium LS10, glycopeptide resistance was transferred to Listeria monocytogenes, Listeria ivanovii, and Listeria welshimeri recipients, whereas no transfer occurred to Listeria seeligeri or Listeria innocua strains. From the two Enterococcus faecalis isolates, no transfer occurred to any Listeria recipient. MICs of both vancomycin and teicoplanin were > or = 256 mg/l for all transconjugants tested. Furthermore, all transconjugants harbored a plasmid that strongly hybridized with the vanA probe, with vanA consistently located in an EcoRI fragment of about 4 kb. Exposure of Listeria transconjugants to vancomycin resulted in synthesis of a membrane protein similar in size (39 kDa) to a vancomycin-induced membrane protein of Enterococcus faecium LS10. In retransfer experiments with Listeria transconjugants used as donors, glycopeptide resistance was transferred to all Listeria recipients tested, including strains of Listeria innocua and Listeria seeligeri, which were unable to receive the resistance from Enterococcus faecium LS10. The frequency of vanA transfer to listerial recipients was greater in retransfer experiments than in the primary matings. These findings suggest that the vanA resistance determinant might spread to the established pathogen Listeria monocytogenes, both directly from a resistant enterococcus and through strains of nonpathogenic Listeria species acting as intermediate resistance vehicles.

Anti-Bacterial Agents↗

PubMed2006

Enterococcus faecalis-mediated biomineralized biofilm formation on root canal dentine in vitro.

Enterococcus faecalis is the most predominant bacteria in teeth with failed root canal therapy and is found to survive harsh conditions prevailing in the root canals of endodontically treated teeth. This study aims to investigate the interaction between E. faecalis and root canal dentine substrate. Towards this end, tooth specimens were prepared and divided into two groups. The tooth specimens in group 1 were incubated with E. faecalis for periods of 2-, 4-, and 6-week intervals and the chemical composition of the biofilm was determined using X-ray diffraction and Fourier transform infrared (FTIR) spectroscopy. The tooth specimens in group 2 were incubated with E. faecalis for a period of 6 weeks and the topography and ultrastructure of the biofilm were examined using scanning electron microscopy (SEM), light microscopy, and laser confocal scanning microscopy. The sediments formed from the bacterial interaction on the dentine (in group 1) were also examined by SEM and FTIR. These experiments highlighted different stages in the interaction of E. faecalis with root canal dentine. Further, a bacterial-induced apatite reprecipitation on mature biofilm was also observed. This ability of E. faecalis to form such calcified biofilm on root canal dentine may be a factor that contributes to their persistence after endodontic treatment.

Biocompatible Materials↗

PubMed2006

Antihypertensive effect of peptides obtained from Enterococcus faecalis-fermented milk in rats.

Previous studies have demonstrated that milk fermented with Enterococcus faecalis decreases the systolic blood pressure (SBP) and the diastolic blood pressure (DBP) of spontaneously hypertensive rats. In this study, we evaluated the antihypertensive activity of the following peptide sequences: LHLPLP, LHLPLPL, LVYPFPGPIPNSLPQNIPP, VLGPVRGPFP, and VRGPFPIIV. These peptides isolated from E. faecalis-fermented milk showed in vitro angiotensin I-converting enzyme-inhibitory activity. Because the most potent angiotensin I-converting enzyme-inhibitory sequences were LHLPLP and LVYPFPGPIPNSLPQ-NIPP, we administered different doses of these peptides to spontaneously hypertensive rats. High doses of the remaining sequences were also administered to these animals. Water served as a negative control and captopril as a positive control. All products were administered orally. The SBP and DBP were measured before administration and also at 2, 4, 6, 8, and 24 h after administration. Before administration of the different products, spontaneously hypertensive rats showed SBP and DBP values of 218 +/- 2.5 and 157 +/- 5.9 mmHg, respectively (n = 30). The sequences LHLPLP, LVYPF-PGPIPNSLPQNIPP, VLGPVRGPFP, and VRGPFPIIV caused clear and significant decreases in SBP, DBP, or both in the animals. In particular, the antihypertensive effect could be clearly established when 2 or 3 mg/kg of LHLPLP was administered. These 2 doses of LHLPLP showed similar antihypertensive properties. Four hours after administration of captopril or the highest doses of the different peptides, the decreases in the SBP and the DBP (mmHg) were as follows: captopril (SBP = 52 +/- 5.8, DBP = 38.8 +/- 3.8), 3 mg/kg of LHLPLP (SBP = 25.3 +/- 8.2, DBP = 29.5 +/- 7.6), 6 mg/kg of LVYPFPGPIP-NSLPQNIPP (SBP = 14.9 +/- 3.7, DBP = 8.7 +/- 4.4), 10 mg/kg of LHLPLPL (SBP = 7.7 +/- 4.1, DBP = 9.4 +/- 3.1), 10 mg/kg of VLGPVRGPFP (SBP = 16.2 +/- 5.8, DBP = 21.64 +/- 3.2), and 10 mg/kg of VRGPFPIIV (SBP = 16.05 +/- 2.74, DBP = 9.19 +/- 3.49). The results obtained suggest that the sequences LHLPLP, LVYPFPGPIPNSLPQ-NIPP, VLGPVRGPFP, and VRGPFPIIV could be responsible, at least in part, for the antihypertensive properties described for E. faecalis-fermented milk.

Animals↗

PubMed1993

Antimicrobial susceptibility of enterococci in vitro.

Treating patients with enterococcal infections has become difficult because of the emergence of several resistant traits in the past decade, including high-level resistance to gentamicin, beta-lactamase production, non-beta-lactamase-associated penicillin resistance and vancomycin resistance. To determine the antimicrobial susceptibility and to survey the antibiotic resistance of enterococci, we tested 111 clinical isolates of enterococci from the National Taiwan University Hospital from 1990 to 1991. There were 102 isolates of Enterococcus faecalis, five of Enterococcus faecium, and four of other enterococcal species. All these isolates were tested in vitro by the agar dilution method for determination of the minimal inhibitory concentration (MIC), by the broth microdilution test for screening of high-level resistance to gentamicin and by the rapid chromogenic cephalosporin method for detection of beta-lactamase production. The effect of the combination of two antimicrobial agents against 20 of the 111 isolates, including strains with high-level and low-level resistance to gentamicin, were further checked by the checkerboard titration method. Among 17 antimicrobial agents, the most active agent was ampicillin, followed by vancomycin, ciprofloxacin, ofloxacin and penicillin. Although none included in these were beta-lactamase producers or vancomycin-resistant strains, there was a high proportion (62%) with high-level resistance to gentamicin among our isolates. There were 11 isolates, seven Enterococcus faecalis and four Enterococcus faecium, which had MICs beyond the clinically achievable level of penicillin (MIC > or = 64 micrograms/mL). Because these resistant isolates of enterococci have important therapeutic implications, periodic surveillance and MIC testing are suggested.

Anti-Bacterial Agents↗

PubMed1999

Comparison of simple and rapid methods for identifying enterococci intrinsically resistant to vancomycin.

Three different methodologies, reduction of litmus milk (LM) and acidification of arabinose (ARA), acidification of methyl-alpha-D-glucopyranoside (MGP), and rapid motility (RM), for differentiating isolates of Enterococcus casseliflavus and Enterococcus gallinarum (intrinsically vancomycin-resistant enterococci [IVRE]) from Enterococcus faecalis and Enterococcus faecium were evaluated. All 33 isolates of E. faecalis tested reduced LM within 4 h and were negative in all other tests, while the 53 isolates of E. faecium were ARA positive only. In contrast, 45 of 46 (98%) IVRE isolates examined (26 E. casseliflavus and 20 E. gallinarum isolates) acidified MGP, 41 of 46 (89%) were LM and ARA positive, and 45 of 46 (98%) were RM positive. Acidification of MGP was therefore the single most useful test for differentiating IVRE from vancomycin-resistant E. faecium and E. faecalis; however, a combination of LM-ARA and RM testing enabled the correct designation of organisms without the need for overnight incubation.

Anti-Bacterial Agents↗

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↗

PubMed1993

NADH binding site and catalysis of NADH peroxidase.

The structure of the complex between cofactor NADH and the enzyme NADH peroxidase from Streptococcus faecalis 10C1 (Enterococcus faecalis) has been determined by crystal soaking, X-ray data collection, model building of NADH and refinement at 0.24-nm resolution based on the known enzyme structure [Stehle, T., Ahmed, S. A., Claiborne, A. & Schulz, G. E. (1991) J. Mol. Biol. 221, 1325-1344]. Apart from NADH, the catalytic center of the enzyme contains FAD and a cysteine that shuttles between thiolate and sulfenic acid states. Unfortunately, this cysteine was irreversibly oxidized to a cysteine sulfonic acid in the established enzyme structure. Based on the geometry of the catalytic center, we discuss the stabilization of the oxidation-sensitive sulfenic acid and propose a reaction mechanism.

Binding Sites↗

PubMed1991

Structure of NADH peroxidase from Streptococcus faecalis 10C1 refined at 2.16 A resolution.

The crystal structure of NADH peroxidase (EC 1.11.1.1) from Streptococcus faecalis 10C1 (Enterococcus faecalis) has been refined to a resolution of 2.16 A using the simulated annealing method. The final crystallographic R-factor is 17.7% for all data in the resolution range 7 to 2.16 A. The standard deviations are 0.015 A in bond lengths and 3.0 degrees in bond angles for the final model, which includes all 447 amino acid residues, one FAD and 369 water molecules. The enzyme is a symmetrical tetramer with point group D2; the symmetry is crystallographic. The redox center of the enzyme consists of FAD and a cysteine (Cys42), which forms a sulfenic acid (Cys-SOH) in its oxidized state. A histidine (His10) close to Cys42 is likely to act as an active-site base. In the analyzed crystal, the enzyme was in a non-native oxidation state with Cys42 oxidized to a sulfonic acid Cys-SO3H. The chain fold of NADH peroxidase is similar to those of disulfide oxidoreductases. A comparison with glutathione reductase, a representative of this enzyme family, is given.

Amino Acid Sequence↗

PubMed2006

Isolation and identification of enterococci from seawater samples: assessment of their resistance to antibiotics and heavy metals.

A hundred Enterococcus strains were isolated from seawater samples collected from coastal areas of Istanbul. Isolates were identified to the species level using standard biochemical tests specified by Facklam and Collins. The species distribution was as follows Enterococcus faecalis (96%), Enterococcus gallinarum (3%) and Enterococcus solitarius (1%). The resistance of bacteria to both heavy metals (zinc [Zn], iron [Fe], cadmium [Cd], chrome [Cr], cobalt [Co]) and antibiotics (ampicillin 10 microg [AP], penicillin G 10 Units [PG], gentamycin 10 microg [GM], streptomycin 10 microg [S], chloramphenicol 10 microg [C], erythromycin 15 microg [E], kanamycin 30 microg [K], amikacin 30 microg [AK], nalidixic acid 30 microg [NA], and vancomycin 30 microg [VA]) was evaluated. None of the strains was resistant to VA. It was found that among the 100 isolates, those that exhibit resistance to antibiotics, particularly NA, S and K, were also resistant all the heavy metals tested. To our knowledge this is the first report focusing on determination of resistance of environmental enterococci found in Istanbul against heavy metals and antibiotics. Thus, combined expressions of antibiotic and heavy metal resistance may help to reinforce ecological and epidemiological studies and to determine the role of these strains in antibiotic and heavy metal resistance dissemination.

Anti-Bacterial Agents↗