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PubMed1998

[Gentamicin resistance of enterococcus genus strains isolated from clinical material].

The occurrence of High Level Gentamicin Resistance (HLGR) was tested among the strains of Enterococcus genus isolated from material potent to microbiological laboratory. The investigation included 184 strains of enterococci, and among them 178 strains of Enterococcus faecalis (96.7%) and 6 strains of Enterococcus faecium (3.3%). HLGR was detected with disks-diffusion method with disks containing 120 micrograms of gentamicin. The screen method with MHA-2 containing 500 mg/l of gentamicin in doubtful results was used. The resistance to high concentration of gentamicin was detected in 29 strains of enterococci (15.76%). The highest percentage of HLGR was proved among strains of enterococci isolated from pyogenic material of hospitalised patients (27.3%).

Anti-Bacterial Agents↗

PubMed2002

[Evaluation of microecology of colonic contents in patients with chronic pancreatitis].

The paper deals with microbiocenosis in the colonic content in 60 patients with chronic pancreatitis during its exacerbation. An analysis has indicated that in the colonic content there are the following microorganisms: Bifidobacterium sp., 10(8)-10(10) CFU/g; Lactobacillus sp., 10(1)-10(5) CFU/g; Escherichia coli, 10(8)-10(10) CFU/g; Enterococcus faecalis, 10(6)-10(10) CFU/g; Enterococcus faecium, 10(6)-10(10) CFU/g; the family Enterobacteriaceae (mainly Klebsiella-Enterobacter sp.), 10(1)-10(6) CFU/g; yeasty (including the Candida genus) and mold fungi, 10(1)-10(5)/g; Staphylococci, 10(1)-10(3) CFU/g. Comparing the findings with those of normal cenosis according to different authors' data suggests that there are no apparently impairments in the levels of some microoranisms, the representives of microbiocenosis in the colonic content in the examinees.

Adolescent↗

PubMed1991

Enumeration by a miniaturized method of Escherichia coli, Streptococcus bovis and enterococci as indicators of the origin of faecal pollution of waters.

Counts of Escherichia coli, faecal streptococci and enterococci were made on faecal specimens from human and animal origin and urban raw sewage waters, with microtiter plates containing selective substances. Escherichia coli was more numerous than faecal streptococci and enterococci in 80% of the samples regardless of the origin. Consequently the use of the ratio E. coli/faecal streptococci to distinguish human from animal origin of faecal pollution is questionable. Enterococcus faecalis was predominant in human and poultry faeces, Streptococcus bovis was typical of the bovine faeces and to a lesser extent also of pig faeces whereas Enterococcus durans, Ent. hirae and Ent. faecium did not characterize any faecal source. Streptococcus bovis could be distinguished in the microtiter plate by its inability to reduce triphenyl tetrazolium chloride (TTC) in the medium.

Animals↗

PubMed2005

Effect of E. faecalis on the release of serine proteases elastase and cathepsin G, and collagenase-2 (MMP-8) by human polymorphonuclear leukocytes (PMNs).

AIM: To measure the release of hydrolytic enzymes [elastase, cathepsin G and collagenase-2 (MMP-8)] from human polymorphonuclear leukocytes (PMNs) during interaction with strains of Enterococcus faecalis isolated from endodontic infections. METHODOLOGY: Six E. faecalis strains isolated from treatment resistant cases of apical periodontitis were included in the study. Overnight cultures of the microbes were used for the experiments. PMNs were isolated using the Ficoll Paque technique, and their vitality was assessed throughout the experiments by the Trypan Blue exclusion test. A known amount of microbes and PMNs were mixed in PBS at +37 degrees C in air, and the release of elastase, cathepsin G and MMP-8 was measured at 0, 20, 60 and 120 min after initiation of incubation. The activities of elastase and cathepsin G were analysed by spectrophotometer assays using specific synthetic peptide substrates, and MMP-8 by western immunoblotting quantitated by computer densitometry. PMNs incubated in buffer without any added microbes served as negative controls, cells incubated with 5 ng mL(-1) phorbol myristic acetate (PMA) served as a positive control. The 95% confidence interval was used to compare the relative amount of elastase and cathepsin G released from the samples. RESULTS: One E. faecalis strain induced a similar or higher elastase, cathepsin G and MMP-8 release than the positive control, whereas the other five strains induced only moderate or no release of the three enzymes examined as compared with the negative and positive controls. Western immunoblot revealed that released MMP-8 had molecular sizes of 60 and 75 kDa representing active and latent forms of MMP-8. In addition, >110 kDa high molecular size and a fragmented 20-30 kDa MMP-8 species could be observed. CONCLUSIONS: The majority of the E. faecalis strains induced little or no release of hydrolytic enzymes from the PMN cells. The finding may partly explain the clinical observation that root canal infections dominated by E. faecalis are usually symptom free.

Blotting, Western↗

PubMed1998

Infective endocarditis presenting as polyarthritis.

We report the case of a patient who complained of arthralgias and arthritis 1 month before the onset of fever or other signs of infective endocarditis. In 2 months she developed an additive, asymmetrical polyarthritis with fever (febrile polyarthritis). Splenomegaly was present. Two-dimensional echocardiography showed no vegetations or other findings suggesting endocardial involvement. Initially, four blood cultures showed no microorganisms, then six of nine subsequent blood cultures grew highly gentamicin-resistant Enterococcus faecalis.

Adult↗

PubMed2005

Antimicrobial activity of daptomycin tested against clinical strains of indicated species isolated in North American medical centers (2003).

Daptomycin is a cyclic lipopeptide approved for use by the US Food and Drug Administration (FDA) for the treatment of complicated skin and skin structure infections caused by Staphylococcus aureus, groups A and B beta-hemolytic streptococci, and vancomycin-susceptible Enterococcus faecalis. We evaluated the daptomycin spectrum against these pathogens by testing 2759 clinical strains consecutively collected in more than 30 hospitals located across the United States and Canada. The isolates were susceptibility tested against daptomycin and many comparators by the reference broth microdilution method. Daptomycin was very active against indicated species with the highest MIC results being 1, 2, and 0.5 microg/mL for S. aureus, E. faecalis, and beta-hemolytic streptococci, respectively. All isolates tested were considered susceptible to daptomycin, according to Clinical and Laboratory Standards Institute and FDA breakpoints, and daptomycin was the most potent (lowest MIC90) among selected antimicrobials commonly used to treat Gram-positive infections. Resistance to oxacillin or vancomycin did not influence daptomycin activity against S. aureus or E. faecalis.

Academic Medical Centers↗

PubMed2007

Comparison of the antimicrobial efficacy of 1.3% NaOCl/BioPure MTAD to 5.25% NaOCl/15% EDTA for root canal irrigation.

The purpose of this study was to compare the antimicrobial efficacy of 1.3% NaOCl/BioPure MTAD to 5.25% NaOCl/15% EDTA for root canal irrigation. Twenty-six bilaterally matched pairs of human teeth were collected. The teeth were incubated with Enterococcus faecalis for 4 weeks. The teeth were divided into two experimental groups and one positive control group. The canals were instrumented and irrigated with either 5.25% NaOCl/15% EDTA or 1.3% NaOCl/BioPure MTAD. Bacterial samples were collected after instrumentation/irrigation and after additional canal enlargement. Statistical analysis of the data using the Wilcoxon Signed Rank test showed significant differences between the experimental groups. The first bacterial samples revealed growth in 0 of 20 samples with 5.25% NaOCl/15% EDTA irrigation and in 8 of 20 samples with 1.3% NaOCl/BioPure MTAD irrigation. Samples taken after additional canal enlargement revealed growth in 0 of 20 samples in 5.25% NaOCl/15% EDTA and in 10 of 20 samples in 1.3% NaOCl/BioPure MTAD group. This investigation showed consistent disinfection of infected root canals with 5.25% NaOCl/15% EDTA. The combination of 1.3% NaOCl/BioPure MTAD left nearly 50% of the canals contaminated with E. faecalis.

Anti-Infective Agents↗

PubMed1994

Evidence for direct-acting oxidative genotoxicity by reduction products of azo dyes.

The intestinal flora forms a complex ecosystem that metabolizes dietary and endogenous nutrients under primarily anaerobic conditions. The ingestion of azo dyes has been proposed as one source of potential genotoxic agents. Many intestinal bacteria are able to reduce the azo bond (termed azofission), which liberates the substituted naphthol compounds. The standard Ames test has not demonstrated mutagenicity either by various common food colorings or by their reduced end products in Salmonella typhimurium strains TA98 and TA100. In contrast, genetic toxicity was demonstrated in the Escherichia coli differential kill assay and in S. typhimurium TA102 for the reduced dyes. The superoxide free radical was produced by the azo dyes only after reduction by the intestinal bacteria Enterococcus faecalis and Bacteroides thetaiotaomicron.

Azo Compounds↗

PubMed1992

Molecular cloning and analysis of the gene encoding the NADH oxidase from Streptococcus faecalis 10C1. Comparison with NADH peroxidase and the flavoprotein disulfide reductases.

The gene encoding the streptococcal flavoprotein NADH oxidase (NOXase), which catalyzes the four-electron reduction of O2-->2H2O, has been cloned and sequenced from the genome of Streptococcus (Enterococcus) faecalis 10C1 (ATCC 11700). The deduced NOXase protein sequence corresponds to a molecular mass of 48.9 kDa and contains three previously sequenced cysteinyl peptides obtained with the purified enzyme. In Escherichia coli, the expressed nox gene produced a catalytically active product, which retained its immunoreactivity to affinity-purified NOXase antisera. Alignment of the NOXase protein sequence with that of streptococcal NADH peroxidase (NPXase) revealed that the proteins are 44% identical. Among the most highly conserved segments is a sequence containing Cys42; this residue is known to exist as a stabilized cysteine-sulfenic acid (Cys-SOH) in NPXase and serves as the non-flavin redox center. In addition, three previously identified NPXase segments, known to be involved in FAD and NAD(P)-binding in other pyridine nucleotide-linked flavoprotein oxidoreductases, are strongly conserved in NOXase. Overall, the extensive homology observed between NOXase and NPXase suggests that the monomer chain fold of the oxidase closely resembles that of the peroxidase. Both sequences share limited but significant homology to those of glutathione reductase and other members of the flavoprotein disulfide reductase family. These and other considerations suggest that these two unusual streptococcal flavoproteins constitute a distinct class of FAD-dependent oxidoreductases, the flavoprotein peroxide reductases, easily contrasted with enzymes such as glutathione reductase and thioredoxin reductase.

Amino Acid Sequence↗

PubMed2002

Distribution of enterococci in Hong Kong.

OBJECTIVES: To study the distribution of hospital isolates of enterococci from urines, bile, blood and body fluids and to evaluate different methods for the identification of enterococci. METHODS: Enterococci isolated from urine, bile, blood and body fluids collected during 1997 and 1998 were identified by polymerase chain reaction (PCR), API 20 Strep and conventional biochemical tests. RESULTS: A total of 498 non-duplicate enterococci were studied: 398 and 43 isolates from urine and bile, respectively, 49 from blood, two from cerebrospinal fluid and six from body fluids. Both API 20 Strep and PCR gave the same identification results for 240 Enterococcus faecalis isolates, 45 E. faecium isolates and one isolate each of E. gallinarum and E. Casseliflavus. These isolates were re-defined by conventional biochemical tests. PCR could correctly identify 303 (98%) isolates while API 20 Strep could only correctly identify 287 (93%) isolates (99% of E. faecalis and 57-87% of the other Enterococcus sp.). Thus, PCR was used in the identification of the remaining isolates and the identity of isolates other than E. faecalis was subsequently confirmed by biochemical tests. CONCLUSIONS: The majority of enterococci isolated was E. faecalis (81%) while only 15% were E. faecium and 4% the other enterococcal species. PCR could correctly identify E. faecalis while the identity of other enterococcal species had to be confirmed by biochemical tests.

Bacterial Typing Techniques↗

PubMed1989

Quality control parameters and interpretive criteria for in vitro susceptibility tests with the macrolide azithromycin. Collaborative Antimicrobial Susceptibility Testing Group.

Quality control parameters for broth microdilution and disk diffusion susceptibility tests were defined and the interpretive criteria for disk diffusion tests reviewed. For interpretation of tests with 15 micrograms azithromycin disks, the following criteria are recommended: greater than or equal to 19 mm for the susceptible category (MIC less than or equal to 2.0 micrograms/ml) and less than or equal to 15 mm for the resistant category (MIC greater than or equal to 8.0 micrograms/ml). Using these criteria, there was 97% overall agreement between broth dilution and disk diffusion tests; Haemophilus influenzae isolates were susceptible to azithromycin by both methods. The quality control strain Staphylococcus aureus ATCC 25923 gave zones of 21 to 26 mm in diameter in a six-laboratory collaborative study. In azithromycin broth microdilution tests the following MIC control limits are recommended: Escherichia coli ATCC 25922, 2.0-8.0 micrograms/ml; Staphylococcus aureus ATCC 29213, 0.25-1.0 micrograms/ml; and Enterococcus faecalis ATCC 29212, 1.0-4.0 micrograms/ml.

Azithromycin↗

PubMed2002

Construction of an integration-proficient vector based on the site-specific recombination mechanism of enterococcal temperate phage phiFC1.

The genome of temperate phage phiFC1 integrates into the chromosome of Enterococcus faecalis KBL 703 via site-specific recombination. In this study, an integration vector containing the attP site and putative integrase gene mj1 of phage phiFC1 was constructed. A 2,744-bp fragment which included the attP site and mj1 was inserted into a pUC19 derivative containing the cat gene to construct pEMJ1-1. E. faecalis KBL 707, which does not contain the bacteriophage but which has a putative attB site within its genome, could be transformed by pEMJ1-1. Southern hybridization, PCR amplification, and DNA sequencing revealed that pEMJ1-1 was integrated specifically at the putative attB site within the E. faecalis KBL 707 chromosome. This observation suggested that the 2,744-bp fragment carrying mj1 and the attP site of phage phiFC1 was sufficient for site-specific recombination and that pEMJ1-1 could be used as a site-specific integration vector. The transformation efficiency of pEMJ1-1 was as high as 6 x 10(3) transformants/microg of DNA. In addition, a vector (pATTB1) containing the 290-bp attB region was constructed. pATTB1 was transformed into Escherichia coli containing a derivative of the pET14b vector carrying attP and mj1. This resulted in the formation of chimeric plasmids by site-specific recombination between the cloned attB and attP sequences. The results indicate that the integration vector system based on the site-specific recombination mechanism of phage phiFC1 can be used for genetic engineering in E. faecalis and in other hosts.

Bacteriophages↗

PubMed2006

Antibiotic resistant fecal isolates of Enterococci among unselected patients outside the clinical sector: an epidemiological study from Southern Germany.

PURPOSE: The aim of the study was to assess the prevalence and determinants of antibiotic-resistant Enterococci in a large group of outpatients in Southern Germany. METHODS: Stool samples were collected from 497 unselected patients aged 40-75 years attending general practitioners. Enterococcus faecium (E. faecium) and Enterococcus faecalis (E. faecalis) were cultured and minimal inhibitory concentrations of antibiotics used inside and outside the clinical sector were tested. RESULTS: E. faecium and E. faecalis could be identified and cultured in 60 (12.4%) and 205 (41.2%) of the stool samples, respectively. Under non-selective culture conditions no vancomycin-resistant Enterococcus (VRE) isolate was found. Only E. faecium isolates showed resistance to fluoroquinolones, 40% were resistant to ciprofloxacin. The prevalences of E. faecium resistance to ampicillin and doxycycline were 3.3% and 13.3%, respectively, whereas 0.5% and 29.6% of the E. faecalis isolates were resistant to ampicillin and doxycycline, respectively. Antibiotic use during the last 3 months was significantly associated with antibiotic resistance (to either ampicillin, imipenem, or doxycycline) of E. faecalis isolates (OR: 2.9; CI: 1.2-6.8). CONCLUSIONS: Prevalences of resistance were generally lower than and patterns of resistance were quite different from previous investigations in the clinical setting. Recent antibiotic use was associated with increased colonization with resistant strains.

Adult↗

PubMed2001

Antibacterial diterpenes from the roots of Salvia blepharochlaena.

The roots of Salvia blepharochlaena have yielded two new diterpenoids, blephaein (1) and O-methylpisiferic acid methyl ester (2), together with eight known diterpenoids. The structures of the new compounds were established by spectroscopic analysis and by some chemical reactions. Potent antibacterial activity was exhibited by the known compounds horminone (9) and 7-acetylhorminone (10) against Staphylococcus aureus ATCC 6538 P, Staphylococcus epidermidis ATCC 12226, and Bacillus subtilis ATCC 6633. Horminone was also found to be active against Enterococcus faecalis ATCC 29212.

Anti-Bacterial Agents↗

PubMed1993

Serological investigation into the association between Streptococcus bovis and colonic cancer.

AIMS: To determine if there was an increase in antibody titre to Streptococcus bovis in patients with colonic cancer, and if this might be a useful marker of the presence for colonic cancer. METHODS: Serum samples from 16 patients and 16 age matched controls were tested by immunoblot and enzyme linked immunosorbent assay (ELISA) against antigen preparations from two strains of S bovis and one strain of Enterococcus faecalis. RESULTS: No distinction between cancer patients and controls could be made using immunoblots. ELISA did show an increase in antibodies to S bovis, but there was a greater increase in antibodies to E faecalis. The increase in antibody titres was greatest with antigens which had been treated with periodate, and was therefore thought not to be caused by antibody to the shared group D carbohydrate antigen. CONCLUSION: It may be possible to construct a test for the detection of colonic cancer based on the detection of antibody to S bovis or E faecalis, though considerable further development of this concept is required.

Antibodies, Bacterial↗

PubMed2006

Biofilm-associated proteins.

Although exopolysaccharides are important and often essential compounds of the biofilm matrix, recent evidences suggest that a group of surface proteins plays a leading role during the development of the microbial communities. The first member of this group of proteins was described in a Staphylococcus aureus bovine mastitis isolate and was named Bap, for biofilm-associated protein. Later on, other surface proteins homologous to Bap and involved in biofilm development have been described in many gram-positive and gram-negative bacteria. In this review, we have summarized our knowledge about three members of this group of proteins: Bap of S. aureus, Esp of Enterococcus faecalis and BapA of Salmonella enterica ser. Enteritidis.

Animals↗

PubMed2000

In vitro yeast infection of human dentin.

An in vitro model was developed for investigation of Candida albicans penetration into human dentinal tubules. The model consisted of a dentin disc mounted between two cuvettes that each had a circular opening facing the disc. The cuvettes were filled with Tryptic-Soy-Broth, and the pulpal side cuvette was inoculated with C. albicans and incubated at 37 degrees C in air until growth occurred in the uninoculated cuvette or up to 30 days. The system was also used with Enterococcus faecalis. Completely glue-covered dentin specimens served as negative controls. Brown & Brenn-stained histological preparations of the specimens were examined with light microscopy. The time needed before growth occurred in the uninoculated cuvette showed great variation with C. albicans, whereas E. faecalis penetrated within 1 to 5 days of incubation. Slight penetration both by hyphae and yeast cells was observed in specimens inoculated with C. albicans, whereas specimens inoculated with E. faecalis showed deep and effective penetration. This study demonstrates the penetration of dentin as a possible pathway of infection by C. albicans. However, dentin penetration by C. albicans was slow and limited in comparison with E. faecalis.

Candida albicans↗

PubMed2003

The antimicrobial effect of MTAD: an in vitro investigation.

Pulp and periradicular diseases are of microbial origin. To effectively clean the root canal system a disinfecting agent must be able to penetrate into difficult-to-reach areas and kill microorganisms with minimal damage to the host tissues. The purpose of this investigation was to test the ability of a mixture of a tetracycline isomer, an acid, and a detergent (MTAD) to kill Enterococcus faecalis and compare its efficacy to that of sodium hypochlorite (NaOCl) and ethylene diamine tetraacetic acid (EDTA). The zones of inhibition and minimum inhibitory concentrations were measured for these solutions. Measurement of zones of inhibition and determination of the minimum inhibitory concentrations showed that MTAD is as effective as 5.25% NaOCl and significantly more effective than EDTA (p < 0.0001). Furthermore, MTAD is significantly more effective in killing E. faecalis than NaOCl when the solutions are diluted (p < 0.0001). Measurement of the minimum inhibitory concentrations demonstrated that although MTAD is still effective in killing E. faecalis at 200x dilution, NaOCl ceases to exert its antibacterial activity beyond 32x dilution. EDTA did not exhibit any antibacterial activity. Based on the results of this study, it seems that MTAD is an effective solution in eradicating E. faecalis.

Chelating Agents↗