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PubMed2004

Antibacterial efficacy of calcium hydroxide, iodine potassium iodide, betadine, and betadine scrub with and without surfactant against E faecalis in vitro.

OBJECTIVE: This study investigated the ability of endodontic irrigants and medicaments to eliminate Enterococcus faecalis from infected dentinal tubules, and whether their antimicrobial action was enhanced by surfactant. STUDY DESIGN: For the study, 5-mm dentin disks were sectioned from bovine incisor roots and infected with E faecalis. Lumens were instrumented, and 1 of 7 medicaments (10% Ca(OH) 2, Betadine, or IKI, each with or without surfactant, or Betadine Scrub) was used to flush and fill each lumen. Positive controls received saline. Specimens were incubated for 15 minutes or 24 hours. Quantitative microbiology of the remaining bacteria was performed and groups were compared using a 1-way ANOVA. RESULTS: The addition of surfactant did not enhance the antibacterial action of any medicament. When used as a 24-hour medicament, Ca(OH) 2 consistently failed to eliminate E faecalis, whereas both Betadine Scrub and IKI rendered 90% of samples sterile. IKI was the only agent shown to consistently eliminate E faecalis in a 15-minute time frame. CONCLUSION: Under the in vitro conditions of this study, IKI was able to eliminate E faecalis from bovine root dentin when used with a 15-minute contact time.

Analysis of Variance↗

PubMed2004

Bacterial resistance to antimicrobials in urinary isolates.

Escherichia coli accounted for about 80% of organisms in uncomplicated urinary tract infections (UTIs), followed by Staphylococcus spp. especially Staphylococcus saprophyticus, and Proteus mirabilis. Against E. coli isolates from patients with uncomplicated UTI, faropenem was the most effective. Up to 1999, fluoroquinolone-resistant isolates were not observed in patients with uncomplicated UTI, but in 2001 fluoroquinolone-resistant E. coli isolates emerged and accounted for about 8%. Various types of organisms were isolated in patients with complicated UTI. Enterococcus faecalis, E. coli, and Pseudomonas aeruginosa were the three most frequent organisms isolated. These three organisms accounted for 44.6%. Amongst oral agents, faropenem showed the lowest rate of resistance against E. coli followed by cephems. The rates of highly fluoroquinolone-resistant and cefpodoxime-resistant E. coli isolates increased rapidly from 1998 to 2001. Fluoroquinolone-resistant P. aeruginosa isolates accounted for about 40% in 2001. Against this species, amikacin was the most effective antimicrobials among all agents tested. About 17% of Pseudomonas were resistant to carbapenem. Eight milligram per litre of ampicillin inhibited all E. faecalis isolates; about 60% of Enterococcus faecium were resistant to ampicillin. The rates of levofloxacin-resistant isolates of E. faecalis and E. faecium were 38 and 97% respectively. UTIs caused by vancomycin resistant enterococci (VRE) are rare in Japan.

Anti-Bacterial Agents↗

PubMed2006

Detection and quantitation of E. faecalis by real-time PCR (qPCR), reverse transcription-PCR (RT-PCR), and cultivation during endodontic treatment.

Enterococcus faecalis is frequently recovered from refractory endodontic infections and has been implicated in endodontic treatment failures. This study compared real-time quantitative PCR (qPCR) assay to cultivation for E. faecalis detection and quantitation during endodontic treatment. A reverse-transcription PCR (RT-PCR) assay was also developed to detect the bacterium clinically in the viable but nonculturable (VBNC) state. Intra-canal samples (n = 87) were collected upon access, post-instrumentation/irrigation and postcalcium hydroxide treatment from 15 primary and 14 refractory infections involving 29 single-rooted teeth, and analyzed by the three methods. The bacterium was up to three times more prevalent in refractory than primary infections at each collection step. Overall, qPCR detected significantly more E. faecalis-positive teeth and samples than cultivation (p < 0.001). VBNC E. faecalis was detected by RT-PCR in four samples that were negative by cultivation. These findings support qPCR and RT-PCR as more sensitive methods than cultivation for detecting E. faecalis in endodontic infections.

DNA, Bacterial↗

PubMed2003

Construction of the mobilizable plasmid pMV158GFP, a derivative of pMV158 that carries the gene encoding the green fluorescent protein.

Plasmid pMV158 has been employed to construct cloning non-mobilizable vectors for various Gram-positive organisms. Here we report the construction of a mobilizable pMV158-based plasmid that harbors the gene encoding the green fluorescent protein under the control of a promoter inducible by maltose. The plasmid was mobilized between strains of Streptococcus pneumoniae as well as from S. pneumoniae to Lactococcus lactis or Enterococcus faecalis at the same frequency as its parental. Transconjugant that received the GFP-tagged plasmid could be detected by their fluorescence, which was especially high in E. faecalis cells.

Conjugation, Genetic↗

PubMed1998

Enterococcal endocarditis--a case treated with teicoplanin and amoxycillin.

The study aimed to determine the antibacterial therapy effective in the cure of endocarditis caused by Enterococcus faecalis resistant to clinically achievable levels of vancomycin. Isolation of the causative enterococcus had been achieved by direct inoculation of the resected valve into the culture medium in theatre. The patient was known to have had an aortic valve defect since childhood and had recently undergone splenectomy following trauma. Blood cultures were negative prior to valve replacement. A perivalvular abscess was noted at operation. In vitro minimal bactericidal results and serum activity were the basis of the postoperative choice of drugs. The minimal bactericidal level of teicoplanin was 250 micrograms/ml and that of amoxycillin 64 micrograms/ml. Neither is achievable with the advocated dosage. A combination of these two cell-wall-active agents successfully eliminated the infection. Acting at two different sites in the synthesis of the bacterial cell wall, teicoplanin and amoxycillin were found to be bactericidal in vitro at the trough levels of the antibiotics in the serum. The patient recovered fully.

Adult↗

PubMed1997

Comparative in-vitro activity of quinupristin/dalfopristin against Enterococcus spp.

The in-vitro activity of quinupristin/dalfopristin against Enterococcus spp. was compared with that of amoxycillin, vancomycin, telcoplanin and erythromycin. The susceptibility of 106 vancomycin-susceptible Enterococcus faecalis, 92 vancomycin-susceptible Enterococcus faecium and 14 vancomycin-resistant enterococci (VRE) was tested. Only one strain of vancomycin-susceptible E. faecium was not susceptible to < or = 0.5 mg/L quinupristin/dalfopristin; this strain required 4 mg/L. All strains of E. faecalis were inhibited by < or = 8 mg/L quinupristin/dalfopristin and all VRE strains were inhibited by < or = 2 mg/L. In contrast, teicoplanin and vancomycin showed inhibitory activity against E. faecalis and E. faecium but not against VRE. Amoxycillin was active against E. faecalis but not usually against E. faecium and showed variable activity against VRE; 38% of E. faecalis, 84% of E. faecium and 80% of VRE strains were resistant to erythromycin. The bactericidal activity of quinupristin/dalfopristin against E. faecium exceeded that of the comparator drugs as judged by MIC:MBC ratios and killing curves. The MBC99 of quinupristin/dalfopristin determined by a microdilution broth technique was < or = 1 mg/L for E. faecium. Four of the five strains of vancomycin-susceptible E. faecium and three of the five VRE strains tested with time-kill curves showed a > or = 2 log reduction in viable count after exposure to quinupristin/dalfopristin for 6 h.

Amoxicillin↗

PubMed2005

Control of Listeria monocytogenes in model sausages by enterocin AS-48.

In this work we describe the control of Listeria monocytogenes CECT 4032 in sausage by adding the enterocin AS-48 producer strains Enterococcus faecalis A-48-32 and Enterococcus faecium S-32-81, and also by adding a semi-purified preparation of the bacteriocin. Addition of preformed AS-48 caused a significant decrease (P<0.01) in the number of viable listeria even at the lowest bacteriocin concentration tested (112 AU/g). At a higher concentration (225 AU/g) listeria were below the detection level (1.99 log units/g) in meat at 3 days of incubation but growth of listeria was observed again after 9 days. For an AS-48 concentration of 450 AU/g, no viable listeria were detected after 6 and 9 days of incubation. When E. faecalis A-48-32 was used as inoculum at approximately 10(7) cfu/g, listeria counts decreased progressively from start of experiment, being below detection level at day 9. The best results were obtained with E. faecium S-32-81, since listeria were undetectable at 6 days of incubation. Bacteriocin concentrations in samples reached concentrations of 60 and 80 AU/g for strains A-48-32 and S-32-81, respectively. These results clearly indicate that AS-48 can be used in the control of L. monocytogenes in sausages.

Animals↗

PubMed1998

Antibiotic-resistant strains of Enterococcus isolated from Swedish and Danish retailed chicken and pork.

Two hundred and seventy-seven Enterococcus isolates representing the predominating enterococcal flora of retailed chicken and pork were identified by phenotypical features, and by random amplified polymorphic DNA. The resistance to nine different antibiotics was determined. Enterococcus faecium was the most frequently occurring species in both Swedish and Danish chicken. Enterococcus faecalis and unidentified groups of Enterococcus dominated in pork. Seventy-three per cent of the Enterococcus isolates from Swedish chicken were resistant to one or more of the tested antibiotics. The corresponding values for Swedish pork, Danish chicken and Danish pork were 9%, 55% and 14%, respectively. Tetracycline resistance was most frequent in isolates from Danish pork and Swedish chicken, while erythromycin resistance was most frequent in Danish chicken. Strains with acquired vancomycin resistance, mainly Ent. faecium, were found only on Danish chicken, except for one isolate from Danish pork. All vancomycin-resistant isolates contained the vanA gene. Vancomycin resistance could be transferred to a vancomycin-sensitive Ent. faecium strain from four of nine tested donor strains.

Animals↗

PubMed1999

Nosocomial bloodstream infections in United States hospitals: a three-year analysis.

Nosocomial bloodstream infections are important causes of morbidity and mortality. In this study, concurrent surveillance for nosocomial bloodstream infections at 49 hospitals over a 3-year period detected >10,000 infections. Gram-positive organisms accounted for 64% of cases, gram-negative organisms accounted for 27%, and 8% were caused by fungi. The most common organisms were coagulase-negative staphylococci (32%), Staphylococcus aureus (16%), and enterococci (11%). Enterobacter, Serratia, coagulase-negative staphylococci, and Candida were more likely to cause infections in patients in critical care units. In patients with neutropenia, viridans streptococci were significantly more common. Coagulase-negative staphylococci were the most common pathogens on all clinical services except obstetrics, where Escherichia coli was most common. Methicillin resistance was detected in 29% of S. aureus isolates and 80% of coagulase-negative staphylococci. Vancomycin resistance in enterococci was species-dependent--3% of Enterococcus faecalis strains and 50% of Enterococcus faecium isolates displayed resistance. These data may allow clinicians to better target empirical therapy for hospital-acquired cases of bacteremia.

Bacteremia↗

PubMed1995

Molecular epidemiology of enterococci with high-level resistance to aminoglycosides.

DNA-based methodologies are considerably more powerful than other phenotype-based typing systems, providing a finer level of epidemiological discrimination, differentiating both closely and distantly related independent isolates that otherwise may appear as identical. In this study, plasmid analysis and pulsed-field gel electrophoresis were used to compare 28 isolates of Enterococci (respectively 13 strains of Enterococcus faecalis and 15 strains of Enterococcus faecium) with high-level resistance to aminoglycosides, isolated in Catania (Italy). Plasmid profile analysis resolved 20 different patterns among 24 plasmid harboring strains; many isolates showed one or two plasmids of the same size, but different plasmid content. Analysis of the PFGE-based RFLP patterns after SmaI digestion of genomic DNA resolved 26 different clones from 28 isolates: particularly, it resolved two different clones from three isolates showing identical plasmid profiles, and it identified as a single clone two isolates exhibiting different plasmid profiles. Thus, on the basis of our PFGE-based RFLP analysis data, we concluded that all the strains included in the study were genetically unrelated with two exceptions.

Aminoglycosides↗

PubMed2000

Susceptibility to quinupristin/dalfopristin and other antibiotics of vancomycin-resistant enterococci from the UK, 1997 to mid-1999.

Susceptibility to quinupristin/dalfopristin and other antibiotics was studied for clinical isolates of vancomycin-resistant enterococci (VRE) referred by UK hospitals between January 1997 and June 1999. Single isolates of VRE from 858 patients in 136 hospitals were received, of which 76% were Enterococcus faecium and 21% were Enterococcus faecalis, the remainder comprising minor species. Most isolates were multi-resistant. After allowing for the effect of blood, which raised the MICs of quinupristin/dalfopristin four-fold, 98.3% of E. faecalis isolates and all the Enterococcus avium, Enterococcus casseliflavus and Enterococcus gallinarum appeared resistant to quinupristin/dalfopristin, whereas 98.8% of the E. faecium isolates and the single Enterococcus raffinosus isolate were susceptible.

Drug Therapy, Combination↗

PubMed2003

Nosocomial bloodstream infections in pediatric patients in United States hospitals: epidemiology, clinical features and susceptibilities.

BACKGROUND: We identified the predominant pathogens and antimicrobial susceptibilities of nosocomial bloodstream isolates in pediatric patients in the US Prospective surveillance for nosocomial bloodstream infections at 49 hospitals during a 6-year period [Surveillance and Control of Pathogens of Epidemiologic Importance (SCOPE)] detected 22 609 bloodstream infections, of which 3432 occurred in patients < or =16 years of age. RESULTS: Gram-positive organisms accounted for 65% of cases, Gram-negative organisms accounted for 24% of cases and 11% were caused by fungi. The overall crude mortality was 14% (475 of 3432) but notably higher for infections caused by Candida spp. and Pseudomonas aeruginosa, 20 and 29%, respectively. The most common organisms were coagulase-negative staphylococci (43%), enterococci, Staphylococcus aureus and Candida spp. (each, 9%). The mean interval between admission and infection averaged 21 days for coagulase-negative staphylococci, 25 days for S. aureus and Candida spp., 32 days for Klebsiella spp. and 34 days for Enterococcus spp. The proportion of methicillin-resistant S. aureus increased from 10% in 1995 to 29% in 2001. Vancomycin-resistance was seen in 1% of Enterococcus faecalis and in 11% of Enterococcus faecium isolates. CONCLUSION: Nosocomial BSI occurred predominantly in very young and/or critically ill children. Gram-positive pathogens predominated across all ages, and increasing antimicrobial resistance was observed in pediatric patients.

Age Distribution↗

PubMed1996

In vitro activities of U-100592 and U-100766, novel oxazolidinone antibacterial agents.

Oxazolidinones make up a relatively new class of antimicrobial agents which possess a unique mechanism of bacterial protein synthesis inhibition. U-100592 (S)-N-[[3-[3-fluoro-4-[4-(hydroxyacetyl)-1-piperazinyl]- phenyl]-2-oxo-5-oxazolidinyl]methyl]-acetamide and U-100766 (S)-N-[[3-[3-fluoro-4-(4-morpholinyl)phenyl]- 2-oxo-5-oxazolidinyl]methyl]-acetamide are novel oxazolidinone analogs from a directed chemical modification program. MICs were determined for a variety of bacterial clinical isolates; the respective MICs of U-100592 and U-100766 at which 90% of isolates are inhibited were as follows: methicillin-susceptible Staphylococcus aureus, 4 and 4 micrograms/ml; methicillin-resistant S. aureus, 4 and 4 micrograms/ml; methicillin-susceptible Staphylococcus epidermidis, 2 and 2 micrograms/ml; methicillin-resistant S. epidermidis, 1 and 2 micrograms/ml; Enterococcus faecalis, 2 and 4 micrograms/ml; Enterococcus faecium, 2 and 4 micrograms/ml; Streptococcus pyogenes, 1 and 2 micrograms/ml; Streptococcus pneumoniae, 0.50 and 1 microgram/ml; Corynebacterium spp., 0.50 and 0.50 micrograms/ml; Moraxella catarrhalis, 4 and 4 micrograms/ml; Listeria monocytogenes, 8 and 2 micrograms/ml; and Bacteroides fragilis, 16 and 4 micrograms/ml. Most strains of Mycobacterium tuberculosis and the gram-positive anaerobes were inhibited in the range of 0.50 to 2 micrograms/ml. Enterococcal strains resistant to vancomycin (VanA, VanB, and VanC resistance phenotypes), pneumococcal strains resistant to penicillin, and M. tuberculosis strains resistant to common antitubercular agents (isoniazid, streptomycin, rifampin, ethionamide, and ethambutol) were not cross-resistant to the oxazolidinones. The presence of 10, 20, and 40% pooled human serum did not affect the antibacterial activities of the oxazolidinones. Time-kill studies demonstrated a bacteriostatic effect of the analogs against staphylococci and enterococci but a bactericidal effect against streptococci. The spontaneous mutation frequencies of S. aureus ATCC 29213 were <3.8 x 10(-10) and <8 x 10(-11) for U-100592 and U-100766, respectively. Serial transfer of three staphylococcal and two enterococcal strains on drug gradient plates produced no evidence of rapid resistance development. Thus, these new oxazolidinone analogs demonstrated in vitro antibacterial activities against a variety of clinically important human pathogens.

Acetamides↗

PubMed2000

Quinupristin-dalfopristin resistance among gram-positive bacteria in Taiwan.

To understand quinupristin-dalfopristin resistance among clinical isolates of gram-positive bacteria in Taiwan, where this agent is not yet available for clinical use, we evaluated 1,287 nonduplicate isolates recovered from January 1996 to December 1999 for in vitro susceptibility to quinupristin-dalfopristin and other newer antimicrobial agents. All methicillin-susceptible Staphylococcus aureus (MSSA) isolates were susceptible to quinupristin-dalfopristin. High rates of nonsusceptibility to quinupristin-dalfopristin (MICs, >/=2 microg/ml) were demonstrated for the following organisms: methicillin-resistant S. aureus (MRSA) (31%), coagulase-negative staphylococci (CoNS) (16%), Streptococcus pneumoniae (8%), viridans group streptococci (51%), vancomycin-susceptible enterococci (85%), vancomycin-resistant Enterococcus faecalis (100%), vancomycin-resistant Enterococcus faecium (66%), Leuconostoc spp. (100%), Lactobacillus spp. (50%), and Pediococcus spp. (87%). All isolates of MSSA, MRSA, S. pneumoniae, and viridans group streptococci were susceptible to vancomycin and teicoplanin. The rates of nonsusceptibility to vancomycin and teicoplanin were 5 and 7%, respectively, for CoNS, ranging from 12 and 18% for S. simulans to 0 and 0% for S. cohnii and S. auricularis. Moxifloxacin and trovafloxacin had good activities against these isolates except for ciprofloxacin-resistant vancomycin-resistant enterococci and methicillin-resistant staphylococci. In Taiwan, virginiamycin has been used in animal husbandry for more than 20 years, which may contribute to the high rates of quinupristin-dalfopristin resistance.

Acetamides↗

PubMed2000

Efficacy of telithromycin (HMR 3647) against enterococci in a mouse peritonitis model.

We used a mouse peritonitis model to evaluate the in vivo efficacy of telithromycin (HMR 3647) (TEL) and erythromycin (ERY) against four strains of Enterococcus faecalis and three strains of Enterococcus faecium with differing susceptibilities to TEL. TEL was highly active in vivo against Ery-susceptible (Ery(s)) and -intermediate (Ery(i)) strains (MIC of TEL = 0.015 to 0.062 microg/ml) and showed less efficacy against Ery-resistant (Ery(r)) isolates (MIC of TEL = 4 to 16 microg/ml), although this was overcome in part by a second subcutaneous dose. Quinupristin-dalfopristin was also noted to have less efficacy against Ery(r) versus Ery(s) or Ery(i) E. faecium strains, but this difference was reduced by intravenous administration. In conclusion, TEL was more potent in vivo against enterococci than was ERY; its activity was lowered by the presence of erm(B)-mediated Ery(r).

Animals↗

PubMed2006

Use of the Phoenix automated system for identification of Streptococcus and Enterococcus spp.

The Phoenix system (Becton Dickinson Diagnostic Systems, Sparks, MD) was evaluated for identification (ID) to the species level of streptococci and enterococci. Two hundred clinical isolates were investigated: beta-hemolytic streptococci (n = 50), Streptococcus pneumoniae organisms (n = 46), viridans group streptococci (n = 31), Enterococcus faecium (n = 36), Enterococcus faecalis (n = 25), and other catalase-negative cocci (n = 12). The API system (bioMérieux, Marcy l'Etoile, France) was used as a comparator. Molecular methods (sequencing of 16S rRNA and zwf and gki genes and ddl gene amplification) were used to investigate discordant results. Upon resolution of discrepancies, correct species ID was achieved by the Phoenix system for 121/129 (93.8%) streptococci and 63/70 (90.0%) enterococci. Excellent results were obtained for S. pneumoniae (45/45) and beta-hemolytic streptococci (49/50). With regard to viridans streptococci, the accuracy of the Phoenix system was 83.9%. Among the latter organisms, the best performance was obtained with isolates of the Streptococcus sanguinis group and Streptococcus anginosus group; problems were instead encountered with the Streptococcus mitis group. Four E. faecium and three E. faecalis isolates were misidentified as Enterococcus casseliflavus/Enterococcus gallinarum or Enterococcus durans. Thus, these isolates were identified only at the genus level. Compared with commercially available systems, the Phoenix system appears a reliable diagnostic tool for identifying clinically relevant streptococci and enterococci. The SMIC/ID-2 panel proved particularly effective for beta-hemolytic streptococci and pneumococci.

Automation↗

PubMedDate not supplied

Increasing prevalence of high-level gentamicin resistance among Enterococci isolated in Greece.

High-level gentamicin-resistant enterococci (HLGRE) (MIC >512 microg/ml) were frequently recovered in the AHEPA University Hospital, Thessaloniki, Greece, during a 15-month period (67/158, 42.4%). PCR testing showed that high-level gentamicin resistance was attributable to the presence of the aac(6')-aph(2") gene, while the recently described genes aph (2")-Ic and aph(2")-Id were not detected. High-level gentamicin resistance could be transferred with varying frequency from 41 of 57 Enterococcus faecalis and 5 of 10 Enterococcus faecium isolates. All transconjugants acquired high-level resistance to streptomycin. HLGRE cause therapeutic problems in our region, since none of the clinically available aminoglycosides can be used in severe infections.

Anti-Bacterial Agents↗

PubMed2005

In vitro activity of daptomycin versus linezolid and vancomycin against gram-positive uropathogens and ampicillin against enterococci, causing complicated urinary tract infections.

OBJECTIVES: The existing therapeutic options for complicated urinary tract infections (UTI) caused by gram-positive uropathogens are not always optimal. Therefore, newer antimicrobials have to be assessed. METHODS: The antimicrobial activity of daptomycin was tested versus linezolid, vancomycin, and ampicillin (enterococci on ly), against pathogens from three different collections: (1) Uropathogens from hospitalized urological patients with complicated and/or hospital-acquired UTIs of the Urologic Clinic, Hospital St. Elisabeth, Straubing. (2) Uropathogens from a multicenter study comprising 37 urological centers throughout Germany. (3) Methicillin-resistant Staphylococcus aureus (MRSA) isolates of patients and staff within the Hospital St. Elisabeth, Straubing. Genotyping of the latter isolates was performed by pulsed-field gel electrophoresis. The minimal inhibitory concentrations (MIC) of daptomycin, linezolid, vancomycin, and ampicillin (only tested against enterococci) were determined by an agar dilution method using a multipointer with an inoculum of 10(4) CFU per point. RESULTS: For all methicillin-susceptible Staphylococcus aureus (n = 25), MRSA (n = 49), methicillin-susceptible coagulase-negative staphylococci (n = 129), methicillin-resistant coagulase-negative staphylococci (n = 33), for Enterococcus faecalis (n = 289), and for Enterococcus faecium (n = 4) the MICs ranged up to 2 mg/l (daptomycin, linezolid), up to 4 mg/l (vancomycin), and up to 8 mg/l (ampicillin, enterococci only) indicating that all strains were susceptible to the antibiotics tested. CONCLUSIONS: According to the in vitro activity daptomycin may be considered a promising antibacterial agent for the treatment of complicated UTI caused by gram-positive uropathogens. Thus, daptomycin should be evaluated in a clinical study.

Acetamides↗