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PubMed1994

Characterization of antibiotic resistance in Streptococcus suis.

Twenty one isolates of Streptococcus suis were screened for antibiotic resistance by growth on antibiotic-containing media, by measuring minimum inhibitory concentrations, by hybridization to specific DNA and oligonucleotide probes for antibiotic resistance genes, and by PCR. The isolates were from a slaughter house survey of respiratory pathogens in Norwegian pigs in 1986. Fifteen isolates were resistant to tetracycline, with MICs ranging from 4-128 micrograms/ml. Genes coding for the Tet O and Tet M determinants were detected in eight and five isolates, respectively. Genes coding for other Gram positive Tet determinants, Tet K, Tet L, and Tet P, were not detected. One isolate was constitutive resistant to erythromycin with MIC of 128 micrograms/ml. Five other isolates carried inducible erythromycin resistance. All these isolates, and five others, were positive in a PCR assay for erythromycin resistance, and hybridized with the Erm C and/or Erm B probes. No resistance against chloramphenicol (5 micrograms/ml) or rifampin (10 micrograms/ml) could be could be detected, but five isolates were resistant to streptomycin (250 micrograms/ml), four isolates were resistant to kanamycin (10 micrograms/ml), and one isolate was resistant to fusicic acid (10 micrograms/ml). In mating experiments with Enterococcus faecalis JH2-2 as recipient, tetracycline, erythromycin, and kanamycin genes were transferred separately to the recipient strain at a rate of 10(-7) transconjugants/recipient cell.

Animals↗

PubMed2004

In vitro evaluation of the effectiveness of irrigants and intracanal medicaments on microorganisms within root canals.

AIM: To evaluate in vitro the effectiveness of sodium hypochlorite (NaOCl), chlorhexidine (CHX) and five intracanal medicaments on microorganisms within root canals. METHODOLOGY: Ninety-six human single-rooted extracted teeth were used. After removing the crowns, canal preparation was completed and the external root surfaces were coated with epoxy resin. Following sterilization, the teeth were contaminated with Candida albicans and Enterococcus faecalis, and were incubated at 37 +/- 1 degrees C for 7 days. The teeth were divided according to the irrigant solution or intracanal medicament: group 1, sterile physiologic solution (SPS) and calcium hydroxide (Ca(OH)2) paste; group 2, SPS and camphorated paramonochlorophenol (CPMC); group 3, SPS and tricresol formalin; group 4, SPS and CaOH2 + CPMC paste; group 5, SPS and PMC furacin; group 6, 2.5% NaOCl without intracanal medication; group 7, 2.0% CHX without intracanal medication and group 8, SPS without intracanal medication (control group). Microbiological samples were collected with sterile paper points, and bacterial growth was determined. The data were submitted to the analysis of variance (anova, P = 0.05). RESULTS: For C. albicans, groups 3 and 8 were statistically less effective than groups 1, 2, 4 and 5 (Kruskal-Wallis (K-W) = 65.241; gl = 7; P = 0.001). For E. faecalis, groups 6 and 8 were statistically less effective than groups 1-4 and 7 (K-W = 61.048; gl = 7; P = 0.001). CONCLUSIONS: Ca(OH)2 + CPMC paste was the most effective intracanal medicament for the elimination of the two microorganisms; 2.0% CHX solution was more effective than 2.5% NaOCl against E. faecalis.

Calcium Hydroxide↗

PubMed2000

Chromosomal integration of the green fluorescent protein gene in lactic acid bacteria and the survival of marked strains in human gut simulations.

An integration vector was constructed to allow introduction of the gfp gene into the chromosomes of Gram-positive bacteria. Integration depends on homologous recombination between a short 458-nt sequence of the tet(M) gene in the vector and a copy of Tn916 in the host chromosome. Strains of Lactococcus lactis IL1403, Enterococcus faecalis JH2-SS, and Streptococcus gordonii DL1 stably marked with single chromosomal copies of the gfp were readily visualised by epifluorescence microscopy. The marked L. lactis strain survived poorly in a continuous culture system inoculated with human faecal flora, while the laboratory E. faecalis strain was lost at approximately the dilution rate of the fermenter.

Bacteria↗

PubMed1991

Relative contributions of host and microbial factors in bacterial translocation.

To study the relative contributions of host and microbial factors in bacterial translocation, germfree mice were mono-associated with either Proteus mirabilis, Escherichia coli, or Enterococcus faecalis. Germfree mice included T-cell-deficient nude mice and normal littermates, natural killer cell-deficient beige mice and normal littermates, and triply immunodeficient mice with beige, T-cell, and B-cell mutations and their littermates. Each bacterial species colonized the cecum in similarly high numbers. Bacteria were recovered from the mesenteric lymph node of every mouse in inconsistent numbers, eg, greater numbers of P mirabilis and E coli were recovered from T-cell-deficient nude mice than from their normal littermates, but the opposite was observed with E faecalis. Comparing the three bacterial species resulted in relatively consistent observations, eg, the incidence of E faecalis translocation to the liver was greater than that of E coli or P mirabilis translocation. Thus, the identity of the translocating microbe significantly affected the recovery of viable translocating bacteria.

Animals↗

PubMed2005

Discovery of a biologically active thiostrepton fragment.

Design, synthesis, and biological evaluation of several domains of the thiopeptide antibiotic thiostrepton led to the discovery of a biologically active fragment. The biological properties of this novel small organic molecule include antibiotic activity against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus faecalis (VREF) bacterial strains, as well as cytotoxic action against a number of cancer cell lines.

Animals↗

PubMed2003

Isolation and structure of palstatin from the Amazon tree Hymeneae palustris(1).

Bioassay (P388 lymphocytic leukemia cell line and human cancer cell lines) guided separation of an extract prepared from the leaves of Hymenaea palustris Ducké led to the isolation of six cancer cell growth inhibitory flavonoids (1-6). The structures were elucidated by HRMS and 1D and 2D NMR spectral analysis. The new flavonolignan 1 designated palstatin proved to be a methoxy structural modification of 5'-methoxyhydnocarpin-D (2). Flavones 1-4 inhibited growth of the pathogenic bacteria Enterococcus faecalis and/or Neisseria gonorrhoeae.

Animals↗

PubMed2002

Infective endocarditis after transrectal prostatic biopsy.

We report the first case of infective endocarditis secondary to transrectal prostatic biopsy, occurring 2 weeks later in a patient with no obvious risk factors. Enterococcus faecalis was isolated in blood and urine cultures. This case suggests that infective endocarditis can be associated with transrectal procedures.

Aged↗

PubMed1995

Incidence of hemolysin, gelatinase, and aggregation substance among enterococci isolated from patients with endocarditis and other infections and from feces of hospitalized and community-based persons.

The presence of hemolysin, gelatinase, and aggregation substance (by use of a probe known to hybridize to most pheromone-responsive plasmids) was determined in 192 isolates of Enterococcus faecalis from patients with endocarditis or other infections and fecal isolates from hospitalized patients or healthy volunteers, and in 86 non-E. faecalis isolates. Hemolysin was more common in nonendocarditis clinical isolates and in hospital fecal isolates (37% and 31%, respectively) than among endocarditis and community fecal isolates (16% and 20%, respectively). Gelatinase and aggregation substance, respectively, were found in 54% and 52% of isolates from endocarditis, in 58% and 72% of isolates from other infections, in 62% and 56% of hospital fecal isolates, and in 27% and 30% of fecal isolates from healthy volunteers. All 86 non-E. faecalis enterococcal isolates were negative for these traits. The absence of hemolysin, gelatinase, or the aggregation substance gene in > 45% of endocarditis E. faecalis isolates suggests that while these traits may play a role in virulence, other properties are also important.

Bacterial Proteins↗

PubMed1997

Use of an excision reporter plasmid to study the intracellular mobility of the conjugative transposon Tn916 in gram-positive bacteria.

An excision reporter plasmid was constructed to characterize the intracellular mobility of Tn916 in various Gram-positive bacteria. The reporter component of this plasmid is a chloramphenicol-resistance gene which has been insertionally inactivated with the integrative vector pAT112 containing the attachment site of Tn916. Tn916-mediated excision of pAT112, to produce clones resistant to chloramphenicol, was detected in Enterococcus faecalis BM4110, Listeria monocytogenes L028-Str and Streptococcus gordonii BM120, but not in Lactococcus lactis MG1363-RF or in Streptococcus pneumoniae BM124, and always depended upon the ability of the bacterial host to generate circular forms of the transposon. The results suggest that (i) the excision event, although required, is not sufficient for conjugal transfer to occur and (ii) there is no linear relationship between the donor potential of E. faecalis strains and either the excision frequency of pAT112 or the copy number of Tn916 circular intermediates per cell in these hosts. Excision of pAT112 occurred mainly during the late exponential phase of growth of E. faecalis and L. monocytogenes and this recombination event was not stimulated by heat shock, salt and alcohol stresses or by the presence of tetracycline in the medium.

Chloramphenicol Resistance↗

PubMed2006

Phenotypic and genotypic identification of enterococci isolated from canals of root-filled teeth with periapical lesions.

The objectives of the present study were to identify enterococcal species isolated from the canals of root-filled teeth with periapical lesions using biochemical and molecular techniques, and to investigate the genetic diversity of the isolates. Twenty-two Enterococcus strains, isolated from the canals of root-filled teeth with persisting periapical lesions, were identified to species level using rapid ID 32 STREP galleries and partial 16S rDNA sequencing. To subtype the strains, genomic DNA from the isolates was analyzed by pulsed field gel electrophoresis (PFGE) after digestion with SmaI. Intragenic regions of two genes, ace and salA, were sequenced for further differentiation of the isolates. All strains were identified as Enterococcus faecalis by both commercial kit and partial 16S rDNA sequencing. PFGE with SmaI of 22 isolates demonstrated 18 macrorestriction profiles, whereas 13 distinct genotypes were identified after analysis of the ace and salA composite sequences. Most of the isolates from distinct patients had different PFGE profiles. Moreover, in two cases, different E. faecalis strains were found in different root-filled teeth from the same mouth. E. faecalis was the only enterococcal species isolated from the canals of root-filled teeth with periapical lesions. Genetic heterogeneity was observed among the E. faecalis isolates following PFGE and sequence-based typing method. Furthermore, the genetic diversity within root canal strains was similar to previous reports regarding E. faecalis isolates from different clinical and geographic origins.

Amino Acid Sequence↗

PubMed1987

Serum bactericidal activity and postantibiotic effect in serum of patients with urinary tract infection receiving high-dose amikacin.

Ten patients received a 30-min infusion of amikacin (30 mg/kg) on day 1 and 15 mg/kg on day 2. Mean serum creatinine was 1.1 +/- 0.3 (standard deviation) mg/dl before and 1.0 +/- 0.3 mg/dl 3 days after the second infusion. Mean serum amikacin concentrations before, at the end of infusion, and 1, 6, 12, and 24 h after 30 and 15 mg/kg were 0, 157, 79, 31, 16, 5, 5, 85, 51, 19, 12, and 5 mg/liter, respectively. Five strains each of Staphylococcus aureus, Staphylococcus epidermidis susceptible and resistant to oxacillin, Streptococcus (Enterococcus) faecalis, corynebacterium sp. strain JK, Listeria monocytogenes, Mycobacterium fortuitum (three strains), Klebsiella pneumoniae, Serratia marcescens, Acinetobacter calcoaceticus, and Pseudomonas aeruginosa were tested. Serum bactericidal activities (SBAs) were greater than or equal to 1:8 in greater than or equal to 80% of the sera 1 and 6 h after 30 mg/kg and in greater than or equal to 60% of the sera 1 and 6 h after 15 mg/kg against Staphylococcus aureus and Staphylococcus epidermidis susceptible to oxacillin, A. calcoaceticus, and K. pneumoniae. L. monocytogenes, Serratia marcescens, and P. aeruginosa had lower SBAs. Very low or no activity was observed against oxacillin-resistant staphylococci and Streptococcus faecalis. The study of the killing rate in serum confirmed these results. Postantibiotic effect was studied by incubating a strain from each species in serum samples obtained 1 and 6 h after both regimens for 0.5, 1, or 2 h. The duration of postantibiotic effect depended on the duration of contact and the concentration of amikacin for the following organisms: oxacillin-susceptible staphylococci, L. monocytogenes, P. aeruginosa, A. calcoaceticus, K. pneumoniae, and Serratia marcescens. M. fortuitum was killed after 30 min of contact. No postantibiotic effect was observed with Streptococcus faecalis, Corynebacterium sp. strain JK, or oxacillin-resistant staphylococci. Amikacin at 30 mg/kg provided high levels and SBAs against susceptible pathogens. Prolonged postantibiotic effects were observed. No signs of nephrotoxicity occurred.

Acinetobacter↗

PubMed1992

Characterization of culturability, protistan grazing, and death of enteric bacteria in aquatic ecosystems.

Nonstained bacteria (NSB), rhodamine-stained bacteria (RSB), and fluorescence-labeled bacteria (FLB) were prepared from two enteric bacterial species, Escherichia coli and Enterococcus faecalis. Counts of CFU of NSB and RSB and total numbers of RSB and FLB were monitored over time, both in the presence and in the absence of natural microbiota. In the presence of natural microbiota, no differences were observed between CFU counts of NSB and RSB, but RSB total numbers were 1 to 4 orders of magnitude higher than CFU numbers. Therefore, the use of standard bacteriological media causes an important underestimation of the total number of enteric bacteria. In the absence of natural microbiota, the total numbers of NSB, RSB, and FLB remained constant over time. These results showed that RSB are a reliable indicator of the decay in both the total number and the CFU of enteric bacteria in natural water samples. By using RSB, enteric bacteria were classified as culturable cells, nonculturable cells (or somnicells), and dead cells in the presence of natural microbiota. In the presence of natural microbiota, differences between RSB and FLB direct counts were detected for E. coli, but not for E. faecalis. These differences were explained by size-selective grazing. Thus, protistan grazing was found to be the main cause of the decrease in total numbers of enteric bacteria in our experiments.

Bacteria↗

PubMed1994

Genetic and molecular studies of a composite chromosomal element (Tn3705) containing a Tn916-modified structure (Tn3704) in Streptococcus anginosus F22.

The plasmid-free Streptococcus anginosus F22 contained a conjugative element, Tn3705, encoding resistance to erythromycin (Emr) and tetracycline-minocycline (Tcr-Mnr). We mapped a chromosomal region (> 52 kb) of F22, corresponding to the internal part of Tn3705. Molecular analysis of Tn3705 revealed it to be a composite structure: it included in its central part a transposon designated Tn3704 (20.3 kb +/- 0.5 kb), which had a modified structure in comparison with that of Tn916 and on which the Emr Tcr-Mn4 markers were localized. Tn3705 inserted from F22 into the chromosome of various streptococcal transconjugants as well as that of Enterococcus faecalis transconjugants without changing its structure. In contrast, from the chromosome of an E. faecalis::Tn3705 transconjugant only Tn3704 inserted, at various sites, into another E. faecalis chromosome. Sugar fermentations occurred after the insertion of Tn3704 into the chromosome of an asaccharolytic E. faecalis strain. Transposition of only Tn3704 from the chromosome of E. faecalis::Tn3705 onto pIP964, an E. faecalis hemolysin plasmid, yielded two different pIP964 derivatives. The size of the entire element Tn3705 was estimated to be about 70.0 kb by pulsed-field electrophoresis.

Chromosomes, Bacterial↗

PubMed2001

Bactericidal properties of the chloroform fraction from rhizomes of Aristolochia paucinervis Pomel.

The deffated chloroform fraction (APRC) obtained from the rhizomes of Aristolochia paucinervis Pomel (Aristolochiaceae) has a high bacteriostatic activity against bacterial strains like Clostridium perfringens ATCC 13124 and Enterococcus faecalis ATCC 29212. Here, we report the bactericidal activity of APRC against both strains which was evaluated by using time-to kill assays. The results showed that APRC produced an intense time-dependent bactericidal effect against C. perfringens, achieving over a 24 h-period a 5log10-unit decrease in CFU/ml at a concentration > or =1.25 x MIC. In contrast, when tested against E. faecalis, APRC exhibited a concentration-dependent killing activity at concentrations of 1.25 x MIC and 2.5 x MIC, yielding to a decrease of 1.5 and 2.5log10-unit in CFU/ml at 4 h, respectively. However, substantial regrowth of E. faecalis occurred within 24 h. Ultrastructural alterations were observed for both exposed microorganisms by scanning and transmission electron microscopy.

Anti-Bacterial Agents↗

PubMed2002

Substantive antimicrobial activity in chlorhexidine-treated human root dentin.

OBJECTIVE: The aim of this in vitro study was to assess the substantive antimicrobial activity of different medicaments in human root dentin. STUDY DESIGN: Canals of 98 roots were enlarged to standard size and medicated for 7 days with the following: (1) 2% chlorhexidine (CHX) gel, (2) 0.2% CHX gel, (3) 2% CHX solution, (4) Ca(OH)(2), (5) Ca(OH)(2)+ 0.2% CHX gel, (6) 2% CHX solution + a 25% CHX-containing controlled-release device, (7) saline, and (8) gel vehicle. After medication, canals were inoculated with Enterococcus faecalis for 21 days. Dentin samples were collected with Gates-Glidden burs into brain heart infusion broth, and bacterial growth was assessed with spectrophotometric analysis of optical density after 72 hours of incubation. RESULTS: Mean optical densities were significantly lower for groups with 2% CHX (1, 3, and 6) when compared with those of the controls (P < .05, analysis of variance with the Tukey test). Other groups did not differ significantly from the controls. CONCLUSIONS: Canal dressing for 1 week with 2% CHX may provide residual antimicrobial activity against E faecalis.

Analysis of Variance↗

PubMed2004

Polymerase chain reaction identification of microorganisms in previously root-filled teeth in a South Korean population.

A large body of evidence indicates that microorganisms are the primary causative agents of endodontic treatment failures. This study intended to assess the occurrence of nine putative endodontic pathogens in root-filled teeth associated with periradicular lesions in a South Korean population using a culture-independent molecular approach. Fourteen root-filled teeth with persistent periradicular diseases were selected for retreatment. After removal of the root canal filling, the canals were sampled, and a polymerase chain reaction assay using taxon-specific oligonucleotide primers was used for microbial detection. Bacteria were present in all cases, as revealed by amplification using ubiquitous 16S rDNA primers. The most frequently detected taxon was Enterococcus faecalis (64%), followed by Streptococcus spp. (21%) and Tannerella forsythensis (14%). The results of this study using a highly sensitive identification method are concurrent with those from other geographical locations using diverse identification methods in that E. faecalis is the main species found in cases of root-filled teeth associated with periradicular lesions.

Adolescent↗

PubMed2000

Carbamate kinase can replace in vivo carbamoyl phosphate synthetase. Implications for the evolution of carbamoyl phosphate biosynthesis.

The exclusive involvement of carbamate kinase (CK) in fermentative ATP production and of carbamoyl phosphate synthetase (CPS) in the production of carbamoyl phosphate (CP) for pyrimidines and arginine biosynthesis was challenged by the finding of CK as the only activity synthesising CP in the archaea Pyrococcus furiosus and Pyrococcus abyssi. We now show that CK can replace CPS in vivo: transformation of Escherichia coli devoid of the CPS gene with plasmids encoding the CK from P. furiosus or from Enterococcus faecalis (which uses CK for making ATP) restores the ability of CPS-deficient E. coli to grow in the absence of arginine and uracil if ammonia and bicarbonate are present.

Ammonia↗

PubMedDate not supplied

Vitrectomy for endophthalmitis after intraocular lens implantation.

PURPOSE: To report the results of vitrectomy and intraocular lens (IOL) removal for the treatment of endophthalmitis after IOL implantation. METHODS: We reviewed 14 eyes of 14 patients who underwent pars plana vitrectomy because of postoperative endophthalmitis. Culture results, surgical methods, and visual outcome are presented. RESULTS: The cultures grew Enterococcus faecalis (n = 3), Staphylococcus epidermidis (n = 2), Propionibacterium acnes (n = 1), and gram-negative bacillus (n = 3). The eyes infected with E. faecalis had poor visual outcome. Eleven eyes treated by the combination of pars plana vitrectomy and IOL removal did not have a recurrence. The remaining 3 eyes on which only vitrectomy was performed had a recurrence, and the additional procedures consisting of vitrectomy and IOL removal could result in eradicating endophthalmitis. CONCLUSIONS: A higher rate of E. faecalis was detected and these eyes had severe inflammation and poor visual outcome. Combined vitrectomy and IOL removal may be a more certain method to prevent recurrence.

Aged↗