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An in vitro investigation of the cariogenic potential of oral streptococci.

Whilst the importance of the mutans streptococci in the aetiology of dental caries is clear, a number of studies have described caries development in their absence. This investigation aimed to assess the cariogenic potential of Streptococcus gordonii, Strep. sanguis, Strep. vestibularis and Enterococcus faecalis in comparison with Strep. mutans and Strep. sobrinus, using a recently described in vitro model. In the presence of a 146 mM sucrose solution and powdered hydroxyapatite, each species was incubated for 5 h, after which acid anion production, calcium release and change in pH were measured. It was possible to assign each species to one of three groups, with E. faecalis, Strep. gordonii, Strep. sanguis exhibiting low, Strep. vestibularis intermediate, and Strep. mutans and Strep. sobrinus high in vitro cariogenic potential. This assay could be used as a screening test to assess the potential cariogenicity of a range of bacterial species.

Acetates↗

Effect of tetracycline on transfer and establishment of the tetracycline-inducible conjugative transposon Tn916 in the guts of gnotobiotic rats.

We have investigated the transfer of Tn916 among strains of Enterococcus faecalis OG1 colonizing in the intestines of gnotobiotic rats. This animal model allows a low limit of detection and efficient colonization of the chosen bacteria. The animals continuously received tetracycline in drinking water. A tetracycline-sensitive recipient strain was allowed to colonize the animals before the resistant donor was introduced. The numbers of donors, recipients, and transconjugants in fecal samples and intestinal segments were estimated. The bioavailable amounts of tetracycline in fecal samples and intestinal segments were monitored by using bacterial biosensors carrying a transcriptional fusion of a tetracycline-regulated promoter and a lacZ reporter gene. Chromosomal locations of Tn916 in transconjugants isolated either from the same animal or from different animals were compared by Southern blot analysis. Our results indicated that selection for the resistant phenotype was the major factor causing higher numbers of transconjugants in the presence of tetracycline. Tetracycline-sensitive E. faecalis cells colonized the intestine even when the concentrations of tetracycline in feces and intestinal luminal contents exceeded growth-inhibitory concentrations. This suggests the existence of tetracycline-depleted microhabitats in the intestinal environment.

Animals↗

Binding properties of Streptococcus gordonii SspA and SspB (antigen I/II family) polypeptides expressed on the cell surface of Lactococcus lactis MG1363.

The oral bacterium Streptococcus gordonii expresses two cell wall-associated polypeptides, designated SspA (1,542 amino acid residues) and SspB (1,462 amino acid residues), that have 70% sequence identity. These polypeptides are members of the antigen I/II family of oral streptococcal adhesins and mediate the binding of streptococci to salivary glycoproteins, collagen, and other oral microorganisms such as Actinomyces naeslundii. To determine if SspA and SspB have differential binding properties, the coding sequences of the sspA and sspB genes were cloned into expression plasmid vector pTREX1-usp45LS to generate pTREX1-sspA and pTREX1-sspB, respectively, and the Ssp polypeptides were displayed on the cell surface of Lactococcus lactis MG1363. Lactococcal cells expressing similar levels of surface SspA or SspB polypeptide were then compared for their abilities to adhere to a range of antigen I/II polypeptide substrates. More than twice as many L. lactis cells expressing SspA bound to immobilized salivary agglutinin glycoprotein (SAG) as did L. lactis cells expressing SspB. In contrast, lactococci expressing SspB adhered twice as well as lactococci producing SspA to collagen type I and to Candida albicans. The binding of A. naeslundii to lactococci was only weakly enhanced by surface expression of Ssp polypeptides. L. lactis(pTREX1-sspB) cells bound in greater numbers to SAG than did Enterococcus faecalis JH2-2 cells expressing SspB from pAM401EB-5. The results suggest that SspA and SspB have markedly different binding affinities for their oral substrates and thus may function to promote site diversity in colonization by S. gordonii.

Adhesins, Bacterial↗

Synthesis and antibacterial evaluation of certain quinolone derivatives.

A number of 7-substituted quinolone derivatives were synthesized and evaluated for antibacterial and cytotoxic activities. Preliminary results indicated that most compounds tested in this study demonstrated better activity against methicillin-resistant Staphylococcus aureus than norfloxacin. Among them, 1-(4-amino-2-fluorophenyl)-6-fluoro-1,4-dihydro-7-[4-[2-(4-methoxyphenyl)-2-hydroxyiminoethyl]-1-piperazinyl]-4-oxo-3-quinolinecarboxylic acid (11d) and its ketone precursor 10d exhibited significant activities against Klebsiella pneumoniae, methicillin-resistant S. aureus, erythromycin- and ampicillin-resistant Streptococcus pneumoniae, and vancomycin-resistant Enterococcus faecalis. Due to strong cytotoxicities of 11d (a mean log GI(50) of -5.40), compound 10d, with good antibacterial activities and low cytotoxicities (a mean log GI(50) of -4.67), is a more potential drug candidate.

Anti-Bacterial Agents↗

Solar inactivation of faecal bacteria in water: the critical role of oxygen.

Suspensions of the faecal indicator bacteria Escherichia coli and Enterococcus faecalis were incubated in full sunlight in plastic bottles containing either (i) air-equilibrated (oxygenated) water or (ii) anaerobic (deoxygenated) water. A rapid decrease in cfu ml-1 was observed for actively growing and stationary phase cells of both types of faecal bacteria when illuminated under aerobic conditions, with Ent. faecalis showing the greater enhancement in the rate of inactivation in air-equilibrated water. The demonstration of an oxygen requirement for the inactivation of faecal bacteria in sunlight indicates that solar-based water disinfection systems are likely to require fully aerobic conditions in order to function effectively.

Aerobiosis↗

Chemomechanical reduction of the bacterial population in the root canal after instrumentation and irrigation with 1%, 2.5%, and 5.25% sodium hypochlorite.

Given the importance of bacteria in the development of periradicular lesions, the eradication of the root canal infection is paramount in endodontic treatment. This study evaluated the in vitro intracanal bacterial reduction produced by instrumentation and irrigation with 1%, 2.5%, and 5.25% sodium hypochlorite (NaOCl) or saline solution. Root canals inoculated with Enterococcus faecalis were instrumented and irrigated with the solutions tested. Canals were sampled before and after preparation. After serial dilution, samples were plated onto Mitis salivarius agar, and the colony-forming units grown were counted. Inhibitory effects of the three NaOCl solutions on E. faecalis were also evaluated by means of the agar diffusion test. All test solutions significantly reduced the number of bacterial cells in the root canal (p < 0.05). There was no significant difference between the three NaOCl solutions tested (p > 0.05). Nonetheless, all NaOCl solutions were significantly more effective than saline solution in reducing the number of bacterial cells within the root canal (p < 0.05). The three NaOCl concentrations showed large zones of inhibition against E. faecalis. The results of this study suggest that regular exchange and the use of large amounts of irrigant should maintain the antibacterial effectiveness of the NaOCl solution, compensating for the effects of concentration.

Analysis of Variance↗

Introduction of Tn916 and pAM beta 1 into Streptococcus bovis JB1 by conjugation.

The transposon Tn916 and self-mobilizing plasmid pAM beta 1 were conjugated from Enterococcus faecalis to the ruminal bacterium Streptococcus bovis JB1. Transconjugants were identified by resistance to tetracycline (Tn916) or erythromycin (pAM beta 1) and by Southern hybridization analyses. Transfer frequencies were 7.0 x 10(-6) and 1.0 x 10(-6) per recipient cell for Tn916 and pAM beta 1, respectively. The transconjugants JB1/Tn916 and JB1/pAM beta 1 were used as donors for matings with E. faecalis, Bacillus subtilis, and the ruminal bacterium Butyrivibrio fibrisolvens. While pAM beta 1 was successfully transferred to all three organisms, Tn916 was transferred only into B. subtilis and B. fibrisolvens at very low frequencies. This is the first report of conjugal DNA transfers between two ruminal organisms.

Conjugation, Genetic↗

Survival of enterococci and Tn916-like conjugative transposons in soil.

The persistence of Enterococcus faecalis, fecal enterococci from swine waste, and Tn916-like elements was determined following inoculation into autoclaved and native soil microcosms. When cells of E. faecalis CG110 (Tn916) were inoculated into native microcosms, enterococcal viability in the soil decreased approximately 5 orders of magnitude (4.8 x 10(5) CFU/g soil to < 10 CFU/g) after 5 weeks. In autoclaved microcosms, the viability of E. faecalis decreased by only 20% in 5 weeks. In contrast, the content of Tn916, based on PCR of DNA extracts from soil microcosms, decreased by about 20% in both native and autoclaved microcosms. Similar results were obtained when the source of fecal enterococci and Tn916-like elements was swine waste. Because the concentration of Tn916-independent E. faecalis DNA (the D-alanine D-alanine ligase gene), based on PCR, decreased to nearly undetectable levels (at least 3 orders of magnitude) after 5 weeks in the native microcosms, the evidence suggests Tn916 stability in the soil results from en masse transfer of the transposon to the normal soil microflora and not survival of E. faecalis DNA in the soil system. Results from denaturing gradient gel electrophoresis suggest that multiple forms of Tn916 occur in swine waste, but only forms most like Tn916 exhibit stability in the soil.

Animals↗

Biocontrol of psychrotrophic enterotoxigenic Bacillus cereus in a nonfat hard cheese by an enterococcal strain-producing enterocin AS-48.

Bacillus cereus is a food poisoning bacterium of great concern, especially in milk products. In this study, we describe the efficient control of the psychrotrophic and toxigenic strain B. cereus LWL1 in milk and in a nonfat hard cow's cheese by the enterocin AS-48 producer strain Enterococcus faecalis A-48-32 (Bac+). No viable B. cereus cells were detected after 72 h incubation in milk coinoculated with the AS-48-producing strain and B. cereus. Diarrheic toxin production was also markedly inhibited by the Bac+ strain to eightfold lower levels compared with control cultures of B. cereus. In cheeses manufactured by inoculation with a commercial starter (about 6.8 log CFU/ml) and B. cereus (about 4 log CFU/ml), the latter reached 6.27 log CFU/g after 5 days of maturation, and approximately 8 log CFU/g after 15 days. However, in cheeses made from milk inoculated with the starter along with a mixture of E. faecalis-B. cereus (2/1 ratio), counts of B. cereus decreased by approximately 1.0, 2.0, 4.32, and 5.6 log units with respect to control cheeses after 5, 10, 15, and 30 days of ripening, respectively. Growth of E. faecalis A-48-32 was associated with enterocin AS-48 production and persistence in cheese. Interestingly, growth of starter cultures was not affected by the Bac+ strain, and neither was lactic acid production. These results clearly indicate that E. faecalis A-48-32 produced satisfactory amounts of bacteriocin in cheese and support the potential use of AS-48-producing strains as culture adjuncts to inhibit B. cereus during cheese manufacture and ripening.

Animals↗

Influence of systemic inflammatory response syndrome on host resistance against bacterial infections.

OBJECTIVE: To determine the relationship between systemic inflammatory response syndrome (SIRS) and host innate immunities against bacterial infections. DESIGN: Controlled animal study. SETTING: University research laboratory. SUBJECTS: Male BALB/c mice, 8-10 wks of age. INTERVENTIONS: Morbidity and mortality rates of severe SIRS mice were compared with those of mild SIRS mice after infection with Enterococcus faecalis or methicillin-resistant Staphylococcus aureus (MRSA) or exposure to infectious complications induced by cecal ligation and puncture (CLP). In addition, a function of effector cells related to antibacterial innate immunities for these infections was analyzed in these two groups. Furthermore, SCIDbgMN mice (SCIDbg mice depleted of antibacterial effector cells) were reconstituted with effector cells from mild or severe SIRS mice and exposed to various infections. MEASUREMENTS AND MAIN RESULTS: Severe SIRS mice were greatly susceptible to E. faecalis, MRSA, and CLP-induced sepsis. On the other hand, as compared with normal mice, mild SIRS mice were resistant to these infections. All of SCIDbgMN mice inoculated with peritoneal macrophages (PMphi) from severe SIRS mice died after infection with E. faecalis or MRSA, whereas all SCIDbgMN mice inoculated with PMphi from mild SIRS mice survived after the same infection. SCIDbgMN mice that were inoculated with PMphi from normal mice and exposed to E. faecalis, MRSA, or CLP survived at rates of 50%, 50%, or 60%, respectively. PMphi from mild SIRS mice exhibited typical properties for classically activated macrophages (CAMphi), whereas those from severe SIRS mice exhibited typical properties for alternatively activated macrophages (AAMphi). CONCLUSIONS: Mphi-associated host antibacterial innate immunities are greatly influenced by SIRS levels. CAMphi, effector cells for the antibacterial innate immunity against E. faecalis, MRSA, and CLP-induced sepsis, are induced in mild SIRS mice. AAMphi with no antibacterial capabilities are generated in mice with severe SIRS. Induction of CAMphi may protect severe SIRS patients against infections.

Analysis of Variance↗

Antibacterial activity of a 1,8-naphthyridine quinolone, PD 131628.

A new 1,8-naphthyridine quinolone antimicrobial, PD131628, was found to be as, or more active than ciprofloxacin or ofloxacin against Escherichia coli and considerably more active than the two 4-quinolone agents against staphylococci in terms of both MIC and OBC (the optimum bactericidal concentration, at which the rate of kill is greatest). In common with ciprofloxacin and ofloxacin, the rate of kill of PD131628 against Enterococcus faecalis was considerably slower than against the other three species tested. Bacterial protein synthesis, RNA synthesis and cell division were not required for the bactericidal activity of PD131628 against E. coli or the staphylococci, although the lethality of PD131628 in the absence of these activities was reduced.

Anti-Infective Agents↗

Culture-independent analysis of indomethacin-induced alterations in the rat gastrointestinal microbiota.

Nonsteroidal anti-inflammatory drugs (NSAIDs) are commonly prescribed for a variety of inflammatory conditions; however, the benefits of this class of drugs are accompanied by deleterious side effects, most commonly gastric irritation and ulceration. NSAID-induced ulceration is thought to be exacerbated by intestinal microbiota, but previous studies have not identified specific microbes that contribute to these adverse effects. In this study, we conducted a culture-independent analysis of approximately 1,400 bacterial small-subunit rRNA genes associated with the small intestines and mesenteric lymph nodes of rats treated with the NSAID indomethacin. This is the first molecular analysis of the microbiota of the rat small intestine. A comparison of clone libraries and species-specific quantitative PCR results from rats treated with indomethacin and untreated rats revealed that organisms closely related to Enterococcus faecalis were heavily enriched in the small intestine and mesenteric lymph nodes of the treated rats. These data suggest that treatment of NSAID-induced ulceration may be facilitated by addressing the microbiological imbalances.

Animals↗

Sufentanil modifies the antibacterial activity of bupivacaine and ropivacaine.

PURPOSE: The purpose of our study was to investigate the effect on the growth of Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), and Enterococcus faecalis (E. faecalis) of bupivacaine at a final concentration of 0.77 mg.mL(-1), ropivacaine at 1.2 mg.mL(-1), and sufentanil at 0.38 and 0.5 microg.mL(-1) (alone or in combination with bupivacaine and ropivacaine). METHODS: The strains were diluted to approximately 3 x 10(4) cfu.mL(-1) in Mueller-Hinton broth. The anesthetics (0.5 mL) were incubated with the bacterial suspensions (0.5 mL) for 24 hr at 37 degrees C. RESULTS: Bupivacaine inhibited the growth of E. coli (59 +/- 0.8%; P < 0.05) and S. aureus (22 +/- 3.6%; P < 0.05). Ropivacaine also inhibited the growth of E. coli (41 +/- 1.2%; P < 0.05) and S. aureus (25.5 +/- 4.1%; P < 0.05). Both anesthetics were ineffective against E. faecalis. Sufentanil only inhibited S. aureus (13.8 +/- 3.1%; P < 0.05) at a concentration of 0.5 microg.mL(-1). Sufentanil modified the antibacterial activity of bupivacaine and ropivacaine. It increased the inhibitory effect of bupivacaine on E. faecalis and S. aureus by 10 +/- 2.1% (P < 0.05) and on E. coli by 7% (P < 0.05). Sufentanil did not increase the inhibitory effect of ropivacaine on the growth of S. aureus. On the other hand, sufentanil reduced the inhibitory effect of ropivacaine on E. coli by 11% (P < 0.05). CONCLUSION: Both bupivacaine and ropivacaine alone or combined with sufentanil inhibited the growth of E. coli and S. aureus. E. faecalis was partially sensitive to a bupivacaine + sufentanil mixture. Sufentanil had a partial synergistic effect on bupivacaine and a partial antagonistic effect on ropivacaine's antibacterial activity.

Amides↗

Antibacterial activity of crude methanolic extract and its fractions of aerial parts of Anthemis tinctoria.

The antibacterial activity of the methanolic extract and its fractions of aerial parts of Aniheinis tinctoria (Asteraceae) was investigated against representative gram-positive Staphylococcus aureus (ATCC 25923) and Enterococcus faecalis (ATCC 29212) and gram-negative strains Escherichia coli (ATCC 25922) and Pseudomonas aeruginosa (ATCC 27853). The activity was concentrated mainly in the dichloromethane (DCM) and hexane fractions of crude methanolic extract. The 5 mg of DCM extract per disk produced 15-16 mm of inhibition zone against S. aureus and P. aeruginosa, however, no activity was found against E. faecalis and E. coli. The hexane fraction showed activity against S. aureus, P. aeruginosa and E. faecalis. As DCM fraction showed the highest antibacterial activity in the disk diffusion assay, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values of only this fraction was determined against S. aureus and P. aeruginosa. These values were found to be in the range of 1.25 to 10 mg/ml.

Anthemis↗

Successful transplantation of organs retrieved from a donor with enterococcal endocarditis.

We report on a patient with an extensive cerebral infarction secondary to enterococcal endocarditis that, in spite of adequate antibiotic treatment, evolved to brain death. The patient was evaluated as a potential organ donor; renal and liver function were normal and both liver and kidneys appeared normal on ultrasonographic examination. When negativity of serial blood and urine cultures was ascertained, liver and kidneys were retrieved for transplantation. The organs were transplanted into three recipients with good results after 14 months of follow-up. All of the recipients received antibiotic prophylaxis against Enterococcus faecalis. None of them has presented infectious complications to date. This case emphasizes that patients with enterococcal endocarditis may be potential organ donors provided both donor and recipients are adequately treated. This is especially important when organs are urgently needed.

Adult↗

Distribution, antibiotic susceptibility and tolerance of bacterial isolates in culture-positive cases of endocarditis in The Netherlands.

During a two-year period data were collected nationwide in The Netherlands on 438 episodes of bacterial endocarditis (BE) in 432 patients. Of the strains isolated in these patients 419 were available for analysis. Of these, 326 were isolated in native valve endocarditis (NVE) and 93 in prosthetic valve endocarditis (PVE). Viridans streptococci, staphylococci and enterococci together constituted 87% of the isolates. More than 46% of the viridans streptococci consisted of Streptococcus sanguis. Enterococcus faecalis and Staphylococcus aureus were the predominant species in the late form of PVE. The majority of the viridans streptococci and haemolytic streptococci were highly susceptible to penicillin. Five of 35 strains of coagulase negative staphylococci were resistant to methicillin. Eleven percent of a random sample of the streptococci collected were tolerant to penicillin. After repeated exposure to a concentration gradient of an appropriate beta-lactam antibiotic, this figure increased to 49%. Of the staphylococci, 5-6% of the strains were tolerant before induction and 16-20% after induction. Of the Enterococcus strains (n = 40), 12.5% showed high-level resistance to one or more aminoglycoside.

Drug Resistance, Microbial↗

Comparative in vitro activity of cefpiramide, a new parenteral cephalosporin.

The in vitro activity of cefpiramide was compared with that of nine other cephalosporins by determining the MIC values by means of an agar dilution method for 300 representative clinical isolates of nonfastidious bacteria. Cefpiramide had a broad-spectrum of activity, similar to that of the third-generation cephalosporins. As judged by the MIC50 and the MIC90 values, cefpiramide was one of the most active cephalosporins against Pseudomonas aeruginosa (MIC50 2.9 micrograms/ml, MIC90 64 micrograms/ml), staphylococci (MIC50 1.2 micrograms/ml, MIC90 10 micrograms/ml) and Enterococcus faecalis (MIC50 13 micrograms/ml, MIC90 43 micrograms/ml). Cefpiramide had moderate activity against Enterobacteriaceae (MIC50 4 micrograms/ml, MIC90 108 micrograms/ml), comparable to that of the older second-generation cephalosporins. Ticarcillin-resistant strains of gram-negative rods were inhibited by higher concentrations of cefpiramide than ticarcillin-susceptible strains.

Bacteria↗

In-vitro comparison of the post-antibiotic effect of vancomycin and teicoplanin.

The post-antibiotic effect (PAE) of teicoplanin was compared with that of vancomycin for five selected Gram-positive cocci. Two concentrations of each antibiotic were investigated, with the test organisms being exposed to each for both 1 and 2 h. At a concentration of 25 mg/l, teicoplanin had a greater PAE than vancomycin for the methicillin resistant staphylococci and Enterococcus faecalis strains tested. At a concentration of 5 mg/l, teicoplanin gave equal effects to those of vancomycin except for the strain of methicillin-resistant Staphylococcus aureus (MRSA) tested. The variation in the antibiotic exposure time gave only small changes in the PAE, except for the MRSA tested.

Enterococcus faecalis↗

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