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[Well water pollution in the Khombole district: research on the contamination by organochlorine pesticide residues and organic substances (feces)].

The study realized in the district of Khombole (SENEGAL) has permit to estimate the contamination levels of wells waters used by the populations. The research and the dosage of the organichlorine pesticide residues, nitrites and nitrates and microbiologic analysis have been done on 19 wells chosen after a drawning of lots. The organochlorine pesticide residues which have been found prove that the wells are permanently exposed to these chemical substances which don't constitute nevertheless a major risk for the populations health. The results of our research proved also that there is a real risk of intoxication with the nitrogen oxides. In effects more than 50% of the wells have revealed nitrates contents up to the indicative value (25 mg/l). As for the nitrates, with a few exceptions (5/18), the contents are superior to the authorized norm (0.1 mg/l). By another way the bacteriologic analysis has revealed in the one hand a DBT (Total Bacterian Count) up to 10,000 germs/l for all the wells, and in the other hand the presence of Escherichia coli, Enterococcus faecalis that confirm a faecal contamination.

Colony Count, Microbial↗

Antibiotic bisanthraquinones produced by a streptomycete isolated from a cyanobacterium associated with Ecteinascidia turbinata.

Chemical studies of a streptomycete isolated from a cyanobacterium associated with the tropical tunicate Ecteinascidia turbinata led to the bioassay-guided purification of two antibacterial bisanthraquinone metabolites and a cytotoxic artifact. The structures, including relative configurations of these octacyclic compounds, were established by spectroscopic analyses. Their potent antibacterial properties (IC(50) = 0.15-130 microM) versus methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus faecalis and cytotoxic effects against HCT-116 cells are presented.

Animals↗

Antimicrobial activity of fusidic acid and disk diffusion susceptibility testing criteria for gram-positive cocci.

The in vitro activity of fusidic acid was assessed and was compared with those of cloxacillin, cefamandole, vancomycin, teicoplanin, ofloxacin, ciprofloxacin, pefloxacin, and fleroxacin against 500 gram-positive cocci: 151 Staphylococcus aureus, 197 coagulase-negative staphylococci, and 152 Enterococcus faecalis strains. All clinical isolates were concomitantly tested by disk diffusion and agar dilution procedures as outlined by the National Committee for Clinical Laboratory Standards. The results with fusidic acid were further analyzed by regression line and error rate-bounded methods. With control American Type Culture Collection organisms, the values were within the limits of the National Committee for Clinical Laboratory Standards or published limits. The incidence of resistance to fusidic acid was 0.7% for S. aureus, 2.5% for coagulase-negative staphylococci, and 99.3% for E. faecalis. The correlation coefficient between the results of disk diffusion and agar dilution tests with fusidic acid was 0.90. Current interpretive criteria for susceptibility to fusidic acid (i.e., MIC of < 2 micrograms/ml and inhibitory zone of 20 mm) gave 1% false susceptibility (all strains being E. faecalis). This error rate is practically eliminated if a zone diameter of 21 mm is considered the breakpoint for susceptibility.

Drug Resistance, Microbial↗

[Antibacterial activity of ofloxacin in urine for 4 days after a single oral dose of 400 mg].

Antibacterial activity of ofloxacin in urine after a single oral dose of 400 mg was evaluated in ten healthy female volunteers. Urine was collected over six periods, i.e., 0-6 h, 6-12 h, 12-24 h, 24-48 h, 48-72 h, and 72-96 h postdose. Ofloxacin levels were assayed in all samples using a microbiological method and HPLC. Urinary ofloxacin MICs were determined for five bacterial strains recovered from urine, two E. coli strains of which one was susceptible and the other resistant to nalidixic acid (Nal-A), one Klebsiella pneumoniae resistant to nalidixic acid (Nal-B), one Staphylococcus saprophyticus strain, and one Enterococcus faecalis strain; MICs were 0.06, 0.25, 1, 0.25, and 2 mg/L, respectively. Mean urinary ofloxacin levels by the microbiological method during the six collection periods were 193.3 +/- 30.3, 138.1 +/- 31, 53.2 +/- 7.3, 8.3 +/- 0.8, 1.4 +/- 0.2, and 0.6 +/- 0.1 mg/L, respectively. HPLC provided similar results: 216.7 +/- 31.6, 130.7 +/- 20.5, 56.5 +/- 7.1, 8.3 +/- 0.9, 1.5 +/- 0.3, and 0.5 +/- 0.05 mg/L, respectively. Mean urinary ofloxacin excretion over 96 h was 67.4 +/- 3.6% of the dose by the microbiological method was 72.5 +/- 2.5% of the dose by HPLC. On the first day, bacteriostatic activity of urine against enterobacteria exceeded 32 and was greater than 8192 for the nalidixic acid-susceptible E. coli strain; on the next day, overall values were equal or greater than 8 for the nalidixic acid-resistant E. coli and K. pneumoniae strains. Bacteriostatic activity was equal to or greater than 32 for the S. saprophyticus strain during the first two days and equal to 8 on the first day and 4 on the second day for the E. faecalis strain.

Administration, Oral↗

Production of bacteriolytic enzymes as a tool for characterizing enterococci.

Bacteriolytic enzymes secreted by log-phase cultures of enterococci (Enterococcus faecalis, Ent. faecium, Ent. durans, Ent. hirae, Ent. casseliflavus, Ent. avium, Ent. mundtii) were analysed by means of a zymogram technique to resolve activities according to the size of their polypeptide component and their specificities towards different substrates. Heterogeneous patterns of lytic activity were observed with different species. For each test substrate, homogeneous patterns of lytic activities were observed with strains of the same species, except for Ent. faecalis strains, which showed heterogeneous lytic patterns even towards the same substrate, and could be divided into at least four different groups according to their lytic pattern. No lytic activity was common to all strains tested. Results of zymogram analysis of Enterococcus bacteriolytic enzymes were consistent with current knowledge on enterococcal taxonomy, indicating that this analytical approach may be a useful tool for fine-tuned characterization of different enterococcal strains.

Bacterial Typing Techniques↗

Identification of a region that influences host range of the streptococcal conjugative plasmid pIP501.

pIP501 is a member of a group of conjugative plasmids that are self-transmissible to a wide variety of streptococci as well as to other gram-positive bacteria. Several pIP501 restriction fragment deletion derivatives have been isolated and characterized. In this paper we describe one such derivative (pVA1702) which was conjugally proficient but had a limited host range. The loss of host range ability was seen as decreased conjugal transfer from Enterococcus faecalis to Streptococcus sanguis and was coincident with the deletion of a 4.5-kb DNA fragment. Transformation of pVA1702 into S. sanguis also was dramatically reduced as compared to its progenitor, suggesting the 4.5-kb fragment encoded a factor(s) necessary for stable maintenance in this host but not in E. faecalis. These observations suggest that pIP501 employs specific mechanisms enabling its maintenance in certain gram-positive bacteria.

Chromosome Deletion↗

Experimental dental caries on gnotobiotic inbred mice.

The purpose of this study in mono-infected gnotobiotic BALB/cA and C3H/HeN mice was to evaluate the cariogenicity of Enterococcus faecalis. The caries incidence and mean caries score in the BALB/cA mice were significantly higher than those in the C3H/HeN. In both of the mouse strains, the mean number of E. faecalis isolated from the cecum content was almost the same, however, the mean number of E. faecalis from the maxilla of BALB/cA was significantly higher than that of C3H/HeN. These results indicate that C3H/HeN has some factors that prevent E. faecalis from attaching to the tooth surfaces.

Animals↗

Transferable erythromycin resistance in Listeria spp. isolated from food.

An erythromycin-resistant (Emr) Listeria innocua and an Emr Listeria monocytogenes isolate both carried ermC genes, which code for rRNA methylases. The ermC genes were transferable by conjugation to recipient L. monocytogenes, Listeria ivanovii, and Enterococcus faecalis but did not appear to be associated with conjugative plasmids.

Chromosomes, Bacterial↗

1H NMR determination of intracellular volume in cell suspensions.

A 1H NMR method has been developed for determining the intracellular and extracellular volumes in a cell suspension. The method is quick, simple, and inexpensive. A comparison of the ratios of the water and Tris buffer resonances in a cell suspension and in a buffer solution gives the intracellular volume. The most important precaution to take is to ensure that coil loading is identical in both solutions and that the NMR signal is not saturating. The method was validated with a 20% polyethylene glycol solution. A comparison with radiolabel methods for volume determination found that the radiolabel probe of extracellular volume did not penetrate the cell wall water of Enterococcus faecalis, resulting in an overestimation of the intracellular volume, and that tritiated water probably exchanged with macromolecules, causing an underestimation of intracellular volume.

Enterococcus faecalis↗

Novel 4-N-substituted aryl pent-2-ene-1,4-dione derivatives of piperazinyloxazolidinones as antibacterials.

A few substituted piperazinylphenyloxazolidinone compounds 6-13 having substitution on the distant nitrogen atom of piperazine ring scaffold have been synthesized and evaluated for their antibacterial activity in Gram-positive bacteria. A few compounds showed superior in vitro antibacterial activity against Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, and Streptococcus pyogenes than linezolid and eperezolid.

Anti-Bacterial Agents↗

Enterococcal plasmid transfer: sex pheromones, transfer origins, relaxases, and the Staphylococcus aureus issue.

Certain conjugative plasmids in Enterococcus faecalis encode a mating response to peptide sex pheromones encoded on the chromosome of potential recipient (plasmid-free) strains. The pheromone precursors correspond to the precursors of surface lipoproteins with the mature peptides coming from the last 7-8 residues of the related signal sequences. Processing that gives rise to the pAD1-related peptide involves a chromosome-encoded metalloprotease (Eep) that is believed to operate within the cytoplasmic membrane. Mutations in the determinants for cAD1 and cAM373, cad and camE, respectively, do not affect cell viability; and when the related plasmid is present, the pheromone response is normal. A cAM373-like activity is produce by Staphylococcus aureus, but the corresponding lipoprotein determinant (camS) is unrelated to the enterococcal determinant (camE). pAD1 has two origins of transfer, oriT1 and oriT2 and encodes a relaxase (TraX), which has been shown to specifically nick in oriT2. pAM373 has a site, oriT, that is similar to oriT2 of pAD1. Both sites (oriT2 of pAD1 and oriT of pAM373) have a series of short direct repeats (5-6 bp with 5-6 bp-spacings) adjacent to a long inverted repeat (140 bp). The direct repeats differ significantly and confer specificity to the two systems. pAD1 and pAM373 are both able to mobilize the nonconjugative plasmid pAMalpha1, which encodes two relaxases that are involved in transfer. Relevant information concerning the possible movement of vancomycin resistance from E. faecalis to S. aureus in a clinical environment is discussed.

Anti-Bacterial Agents↗

Trehalose-6-phosphate phosphorylase is part of a novel metabolic pathway for trehalose utilization in Lactococcus lactis.

Lactococcus lactis splits phosphorylated trehalose by the action of inorganic phosphate-dependent trehalose-6-phosphate phosphorylase (TrePP) in a novel catabolic pathway. TrePP was found to catalyze the reversible conversion of trehalose 6-phosphate into beta-glucose 1-phosphate and glucose 6-phosphate by measuring intermediate sugar phosphates in cell extracts from trehalose-cultivated lactococci. According to native PAGE and SDS-PAGE, TrePP was shown to be a monomeric enzyme with a molecular mass of 94 kDa. Reaction kinetics suggested that the enzyme follows a ternary complex mechanism with optimal phosphorolysis at 35 degrees C and pH 6.3. The equilibrium constants were found to be 0.026 and 0.032 at pH 6.3 and 7.0, respectively, favoring the formation of trehalose 6-phosphate. The Michaelis-Menten constants of TrePP for trehalose 6-phosphate, inorganic phosphate, beta-glucose 1-phosphate, and glucose 6-phosphate were determined to be 6, 32, 0.9, and 4 mm, respectively. The TrePP-encoding gene, designated trePP, was localized in a putative trehalose operon of L. lactis. This operon includes the gene encoding beta-phosphoglucomutase in addition to three open reading frames believed to encode a transcriptional regulator and two trehalose-specific phosphotransferase system components. The identity of trePP was confirmed by determining the N-terminal amino acid sequence of TrePP and by its overexpression in Escherichia coli and L. lactis, as well as the construction of a lactococcal trePP knockout mutant. Furthermore, both TrePP and beta-phosphoglucomutase activity were detected in Enterococcus faecalis cell extract, indicating that this bacterium exhibits the same trehalose assimilation route as L. lactis.

Amino Acids↗

Dentinal tubule disinfection using three calcium hydroxide formulations.

The objective of this study was to determine the antibacterial efficacy of three calcium hydroxide (CH) formulations using an in vitro model of Enterococcus faecalis dentinal tubule infection. CH mixed with water (CH), CH mixed with iodine-potassium iodide (CH+IKI), and CH mixed with iodoform and silicone oil (Metapex) were tested. Human cylindrical dentin specimens infected with E. faecalis were filled with disinfectants and incubated for 1 week. Dentin powder samples collected with ISO 018 burs showed a statistically significant reduction in E. faecalis for all three experimental groups in comparison with untreated control specimens. Statistically significant differences were also found between the three experimental groups. Metapex was the most effective dentinal tubule disinfectant, followed by CH+IKI and then CH. Similar results were observed at greater dentin tubule depths (ISO 021 burs) with the exception that intracanal treatment with CH resulted in significantly higher numbers of E. faecalis in comparison with untreated control specimens.

Anti-Infective Agents, Local↗

Antimicrobial efficacy of ozonated water, gaseous ozone, sodium hypochlorite and chlorhexidine in infected human root canals.

AIM: To determine the antimicrobial efficacy of ozonated water, gaseous ozone, sodium hypochlorite and chlorhexidine in human root canals infected by Enterococcus faecalis. METHODOLOGY: Thirty human maxillary anterior teeth were prepared and inoculated with E. faecalis for 60 days. Eppendorf tubes were connected to the coronal portion of the teeth. Urethane hoses were attached to the tubes and to the entrance of a peristaltic pump. The exit of the apparatus corresponded to the apical portion of the root canals. The test irrigating solutions were ozonated water, gaseous ozone, 2.5% sodium hypochlorite (NaOCl), 2% chlorhexidine that circulated at a constant flow of 50 mL min(-1) for 20 min. Samples from the root canals were collected and immersed in 7 mL Letheen Broth (LB), followed by incubation at 37 degrees C for 48 h. Bacterial growth was analysed by turbidity of the culture medium and subculture on a specific nutrient broth. A 0.1 mL inoculum obtained from LB was transferred to 7 mL of brain heart infusion and incubated at 37 degrees C for 48 h. Bacterial growth was checked by turbidity of the culture medium carried out in triplicate. RESULTS: No solution used as an irrigant over a 20-min contact time demonstrated an antimicrobial effect against E. faecalis. CONCLUSION: The irrigation of infected human root canals with ozonated water, 2.5% NaOCl, 2% chlorhexidine and the application of gaseous ozone for 20 min was not sufficient to inactivate E. faecalis.

Anti-Infective Agents, Local↗

The antimicrobial effect of calcium hydroxide in root canals pretreated with 5% iodine potassium iodide.

Calcium hydroxide (CH) is often used as a routine interappointment dressing during the endodontic treatment of teeth with apical periodontitis. However, it fails to consistently produce sterile root canals. The present study was set up to find out whether an antimicrobial strategy including the use of CH could be made more effective if: 1) canals were pretreated with 5% iodine potassium iodide (IPI), and 2) the dressing period was extended up to 2 months. Fifty human teeth, with radiographically verified apical periodontitis, were microbiologically sampled. After chemomechanical preparation the canals were pretreated with IPI for 3-7 days. Teeth where microorganisms persisted were then treated with CH for 2 months. Following instrumentation and dressing with IPI, 43 bacterial strains were recovered in 22 of the teeth. Samples obtained after the CH dressing period disclosed growth of 13 facultative and two strict anaerobic strains in 10 teeth. Enterococcus faecalis was identified in two specimens. In conclusion, the present study gave no evidence for an increased antimicrobial effect of CH if it was left for longer periods in the root canal. Although pretreatment with IPI from a quantitative point of view did not seem to add antimicrobial power, it might reduce the frequency of persisting strains of E. faecalis.

Adult↗

Renal pharmacokinetic changes of gentamicin during enterococcal pyelonephritis.

In the present study, the renal pharmacokinetics of gentamicin was investigated in pyelonephritic rats infected with Streptococcus (Enterococcus) faecalis. Four days after the induction of infection, animals were given either a single dose of gentamicin or two daily injections for 7 days. The treated animals were evaluated from 1 h to 6 months posttreatment. After a single injection, limited pharmacokinetic variations were observed, whereas after 14 injections infected kidneys demonstrated significantly higher concentrations and a more extended renal elimination phase of the antibiotic. Analysis of the area under the curve of the concentration in kidney versus time showed more marked renal accumulation by the infected and long-term treated animals than by normal animals or those receiving only one injection of aminoglycoside. Renal function remained normal in both the infected and normal animals treated with this aminoglycoside. These results demonstrate that S. faecalis pyelonephritis disturbs the renal handling of gentamicin and may increase the susceptibility of the kidney to aminoglycosides.

Animals↗

Pharmacodynamic interaction between RP 59500 and gram-positive bacteria infecting fibrin clots.

The fibrin clot penetration and in vivo bactericidal activity of RP 59500, a new semisynthetic streptogramin, for two Staphylococcus aureus strains (one methicillin resistant and the other methicillin susceptible), two Staphylococcus epidermidis strains (one methicillin resistant and the other methicillin susceptible), and one Enterococcus faecalis strain were evaluated. The clots, inserted subcutaneously, were infected with a mean of 10(8) CFU of the pathogen per g. For each strain, groups of four rabbits received a single intravenous injection of 50 mg of RP 59500 per kg of body weight over 30 min. The mean peak level of RP 59500 in serum in the infected rabbits was 61.9 +/- 6.3 micrograms/ml. The drug was detectable in serum at a level of 0.8 micrograms/ml up to 4 h after administration. The mean peak fibrin clot drug level at 1 h was 3.3 +/- 0.1 micrograms/g. At 6 h, the level in clots was 1.2 +/- 0.1 micrograms/g. The mean half-life in serum in infected rabbits was 0.34 +/- 0.01 h, while in clots the drug exhibited a longer half-life of 3.8 +/- 0.4 h. In vivo, this new streptogramin sterilized the clots infected with the two S. aureus strains studied in less than 1 h and induced a marked reduction in colony counts of the two S. epidermidis strains studied for up to 24 h. The activity of the streptogramin against E. faecalis was limited. These results suggest that RP 59500 should be further evaluated for the treatment of infection with methicillin-resistant staphylococci.

Animals↗