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Previous studies have demonstrated the acrylamide-removing properties of probiotic monocultures; however, potential advantages of consortia over monocultures in reducing the dietary exposure to acrylamide have not been proven. Hence this work aims to assess the acrylamide (AA)-binding properties of bacterial consortia, consisting of either probiotic strains and / or representative bacteria of duodenal microbiota, exposed to simulated gastrointestinal conditions (SGC). The AA binding capacity of ten probiotic strains (PS) and six duodenal strains (NDS) was evaluated under different conditions; then, three different consortia (PS, NDS, and PS + NDS) were assessed under SGC.Among individual PS, Bacillus coagulans GBI-30, Lactobacillus fermentum J23, L. pentosus J37 and J24, and L. casei Shirota, exhibited the highest AA-binding capacity (80-87%), while Bifidobacterium catenulatun ATCC27676, Streptococcus salivarius subsp. thermophilus ATCC19258, and S. gallolyticus ATCC9809 were the best (ca. 68%) NDS monocultures. Probiotic strain consortia showed higher (P 0.05) AA binding capacity ( 90%) than monoculture bacteria. Conversely, individual NDS cultures displayed higher (P 0.05) binding capacity than NDS consortia (60%). A significant reduction (P 0.05) in AA removal capacity was observed when consortia were exposed to SGC, PS consortia being the most effective (60% removal).These results suggest that consortia of specific PS could play an important role in reducing the intestinal availability of acrylamide. © 2021 Society of Chemical Industry.

Background The aim of the study was to observe the antimicrobial activity of Porphyromonas gingivalis and Treponema denticola as well as the effect on reducing volatile sulfur compounds (VSCs). Materials and methods After P. gingivalis and T. denticola were cultured with or without Streptococcus salivarius K12 and M18, VSCs were measured by Oral Chroma. In order to analyze the mechanism for malodor control, the antimicrobial activity of S. salivarius K12 and M18 against P. gingivalis and T. denticola was assessed. SPSS 21.0 was used for data analysis with the Kruskal–Wallis and Jonckheere–Terpstra tests. Mann–Whitney test was applied for post hoc analysis. Results P. gingivalis and T. denticola VSC levels were reduced by high concentrations of S. salivarius K12 and M18 during coculture. The concentrations were lower than those of single culture (p < .05). An antimicrobial effect was detected on P. gingivalis, and T. denticola by 50% S. salivarius K12 and M18. The spent culture medium and whole bacteria of S. salivarius K12 and M18 reduced the levels of VSCs below the amount in a single culture of P. gingivalis and T. denticola (p < .05). Conclusion S. salivarius K12 and M18 decreased the levels of VSCs originating from P. gingivalis and T. denticola.

The members of the infant microbiome are governed by feeding method (breastmilk vs. formula). Regardless of the source of nutrition, a competitive growth advantage can be provided to commensals through prebiotics - either human milk oligosaccharides (HMOs) or plant oligosaccharides that are supplemented into formula. To characterize how prebiotics modulate commensal - pathogen interactions, we have designed and studied a minimal microbiome where a pathogen, Streptococcus agalactiae engages with a commensal, Streptococcus salivarius. We discovered that while S. agalactiae suppresses the growth of S. salivarius via increased lactic acid production, galacto-oligosaccharides (GOS) supplementation reverses the effect. This result has major implications in characterizing how single species survive in the gut, what niche they occupy, and how they engage with other community members.

AIM To investigate whether glycosaminoglycans (GAGs) binding to high-dose LL37 eliminates its cytotoxicity to dental pulp cells (hDPCs) whilst retaining undiminished antimicrobial and LPS-neutralizing abilities. METHODOLOGY hDPCs were stimulated with varying concentrations of LL37, and their cell viability was analysed by MTT. Then, high-dose LL37 (10 μmol L-1 ) was bound to varying concentrations of three GAGs, heparin, chondroitin sulphate and hyaluronic acid, and their cytotoxic effects on hDPCs and antimicrobial effects were evaluated and compared. Furthermore, the LPS-neutralizing ability of heparin (5 μg mL-1 )-LL37 (10 μmol L-1 ) complexes, which were found to be less cytotoxic for hDPCs with undiminished antimicrobial ability, was investigated. Statistical analysis was performed using one-way analysis of variance (anova), followed by Dunnett's test. P values below 0.05 were considered significant. RESULTS LL37 significantly reduced the cell viability of hDPCs in a dose-dependent manner (P

SUMMARY Bacterial adhesion is a critical step for colonization of the host. The pioneer colonizer and commensal bacterium of the human gastrointestinal tract, Streptococcus salivarius, has strong adhesive properties but the molecular determinants of this adhesion remain uncharacterized. Serine-rich repeat (SRR) glycoproteins are a family of adhesins that fulfill an important role in adhesion. In general, Gram-positive bacterial genomes have a unique SRR glycoprotein-encoding gene. We demonstrate that S. salivarius expresses three large and glycosylated surface-exposed proteins — SrpA, SrpB and SrpC — that show characteristics of SRR glycoproteins and are secreted through the accessory SecA2/Y2 system. Two glycosyltransferases — GtfE/F — encoded outside of the secA2/Y2 locus, unusually, perform the first step of the sequential glycosylation process, which is crucial for SRR activity. We show that SrpB and SrpC play complementary adhesive roles involved in several steps of the colonization process: auto-aggregation, biofilm formation and adhesion to a variety of host epithelial cells and components. We also show that at least one of the S. salivarius SRR glycoproteins is important for colonization in mice. SrpA, SrpB and SrpC are the main factors underlying the multifaceted adhesion of S. salivarius and therefore play a major role in host colonization. This article is protected by copyright. All rights reserved.

Objectives/Hypothesis Silicone voice prostheses are most frequently used in voice rehabilitation of laryngectomized patients. However, the functional device lifetimes are limited due to formation of mixed biofilms. Existing in vitro models simulating biofilm formation are restricted to only short-term periods. Study Design The goal of this study was to determine the effect of carboxymethyl chitosan on mixed biofilm formation of fungi and bacteria on silicone over a long-term period. Methods Mixed species biofilms of Candida albicans, Candida tropicalis, Lactobacillus gasseri, Streptococcus salivarius, Rothia dentocariosa, and Staphylococcus epidermidis were cultivated on the surfaces of medical-grade silicone with and without addition of carboxymethyl chitosan. Biofilm kinetics was monitored using specially designed image analysis software to calculate the percentual surface covering of each platelet. Biofilm architecture was investigated by scanning electron microscopy. Results A cover of living mixed biofilm could be generated over 22 days on silicone and the maximum of 22% biofilm surface covering at day 22. However, less than 4% surface coverage was observed on the carboxymethyl chitosan-treated plates in the testing period. Scanning electron microscopy confirms that, on surfaces treated by carboxymethyl chitosan, the biofilm was less dense. In addition, there were fewer layers of cells and profuse cellular debris, together with degrading and morphologically altered yeast cells. Conclusion Carboxymethyl chitosan may serve as a possible antibiofilm agent to limit biofilm formation on voice prostheses. Level of Evidence N/A. Laryngoscope, 2016

Rationale Chronic kidney disease (CKD) and periodontitis (PD) are important health issues. There is a large variety of microorganisms related to the pathogenesis of periodontitis, and optimising the time and the cost of laboratory assays to detect these organisms is highly valuable in the medical field. Methods Bacteria were isolated from saliva and oral biofilm of 30 adolescents and young adults with definite medical and dental diagnosis of CKD and PD, respectively, and proteins were extracted for microorganism identification by means of the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) technique. Results The results showed that the most incident microorganisms were Actinomyces dentalis (43%), Acinetobacter ursingi (60%), Aggregatibacter actinomycetencomitans (60%), Corynebacterium argentoctens (63%), Staphylococcus aureus (93%), Streptococcus salivarius (97%) and Tannerella forsythensis (43%). The analysis of oral biofilm showed higher incidences for Actinomyces dentalis (33%), Acinetobacter ursingi (50%), Aggregatibacter actinomycetencomitans (50%), Corynebacterium argentoctens (70%), Pseudomonas aeruginosa (40%), Staphylococcus aureus (73%) and Streptococcus salivarius (87%). Conclusions Based on these results, we concluded that the MALDI Biotyper protocol proves useful as a rapid and reliable assay for distinguishing different microorganisms possibly related to CKD and PD. Copyright © 2016 John Wiley & Sons, Ltd.

We report streptococcal dysbiosis in acute diarrhoea irrespective of aetiology. Compared with 20 healthy local controls, 71 Bangladeshi children hospitalized with acute diarrhoea (AD) of viral, mixed viral/bacterial, bacterial and unknown aetiology showed a significantly decreased bacterial diversity with loss of pathways characteristic for the healthy distal colon microbiome (mannan degradation, methylerythritol phosphate and thiamin biosynthesis), an increased proportion of faecal streptococci belonging to the Streptococcus bovis and Streptococcus salivarius species complexes, and an increased level of E. coli-associated virulence genes. No enteropathogens could be attributed to a subgroup of patients. Elevated lytic coliphage DNA was detected in 2 out of 5 investigated enteroaggregative E. coli (EAEC)-infected patients. Streptococcal outgrowth in AD is discussed as a potential nutrient-driven consequence of glucose provided with oral rehydration solution. ispartof: ENVIRONMENTAL MICROBIOLOGY vol:20 issue:6 pages:2256-2269 ispartof: location:England status: published

As a highly prevalent multifactorial disease, dental caries afflicts a large proportion of the world's population. As teeth are constantly bathed in saliva, the constituents and properties of this oral fluid play an essential role in the occurrence and progression of dental caries. Various inorganic (water and electrolytes) and organic (proteins and peptides) components may protect teeth from dental caries. This occurs via several functions, such as clearance of food debris and sugar, aggregation and elimination of microorganisms, buffering actions to neutralize acid, maintaining supersaturation with respect to tooth mineral, participation in formation of the acquired pellicle and antimicrobial defense. Modest evidence is available on the associations between dental caries and several salivary parameters, including flow rate, buffering capacity and abundance of mutans streptococci. Despite some controversial findings, the main body of the literature supports an elevated caries prevalence and/or incidence among people with a pathologically low saliva flow rate, compromised buffering capacity and early colonization or high titer of mutans streptococci in saliva. The evidence remains weak and/or inconsistent on the association between dental caries and other saliva parameters, such as other possible cariogenic species (Lactobacillus spp., Streptococcus sanguis group, Streptococcus salivarius, Actinomyces spp. and Candida albicans), diversity of saliva microbiomes, inorganic and organic constituents (electrolytes, immunoglobulins, other proteins and peptides) and some functional properties (sugar clearance rate, etc.). The complex interactions between salivary components and functions suggest that saliva has to be considered in its entirety to account for its total effects on teeth.

The inhibition of bacterial growth through effective non-toxic antimicrobial substances is of great importance for the prevention and therapy of implant infections in various medical disciplines. For the evaluation of a therapeutic window of silver nanoparticles (AgNPs), their bactericidal properties were tested in agar composites and colloids on four medical relevant bacteria. Therefore, we produced AgNPs using high-power nanosecond laser ablation in water showing a log-normal particle diameter distribution centered at 17 nm. Bacteria were incubated with AgNP concentrations ranging from 5 to 70 µg · mL−1 and the growth rate was recorded. Additionally, cytotoxic effects of AgNPs on human gingival fibroblasts were examined. The experiments demonstrated that laser-synthesized AgNPs resulted in a significant bacterial growth inhibition of more than 80% at the indicated concentrations in a solid agar model (Pseudomonas aeruginosa 10 µg · mL−1, Streptococcus salivarius 10 µg · mL−1, Escherichia coli 20 µg · mL−1, Staphylococcus aureus 70 µg · mL−1). In a planktonic bacteria model, the growth of the tested bacteria was significantly delayed by the addition of AgNPs at a concentration of 35 µg · mL−1. The cytotoxic assays showed limited adverse effects on human fibroblasts at concentrations of less than 20 µg · mL−1. The present study illustrates the strong antibacterial effects of ligand-free, laser-generated AgNPs that exhibit moderate cytotoxic effects, resulting in a therapeutically applicable concentration of AgNPs for medical purposes between 10 and 20 µg · mL−1.

International Journal of Paediatric Dentistry 2011; 21: 401–406 Background. Early in life, vaginally delivered infants exhibit a different composition of the gut flora compared with infants delivered by caesarean section (C-section); however, it is unclear whether this also applies to the oral cavity. Aim. To investigate and compare the oral microbial profile between infants delivered vaginally and by C-section. Design. This is a cross-sectional case–control study. Eighty-four infants delivered either vaginally (n = 42) or by C-section (n = 42) were randomly selected from the 2009 birth cohort at the County Hospital in Halmstad, Sweden. Medically compromised and premature children (

In the present work, the anticariogenic activities of nine labdane type-diterpenes and four sesquiterpenes were investigated. Among these metabolites, (-)-copalic acid (CA) was the most active compound displaying MIC values very promising (ranging from 2.0 to 6.0 µg/mL) against the main microorganisms responsible for dental caries: Streptococcus salivarius, S. sobrinus, S. mutans, S. mitis, S. sanguinis and Lactobacillus casei. Time kill assays performed with CA against the primary causative agent (S. mutans) revealed that, in the first 12 h, this compound only inhibits the growth of the inoculum (bacteriostatic effect). However, its bactericidal effect is clearly noted thereafter (between 12 and 24 h). Also, CA did not show a synergistic effect when combined with the anticariogenic gold standard (chlorhexidine, CHD) in the checkerboard assays against S. mutans. In conclusion, the results points out CA as an important metabolite in the search for new effective anticariogenic agents. Copyright © 2010 John Wiley & Sons, Ltd.

Summary Oral viridans group streptococci are the major commensal bacteria of the supragingival oral biofilm and have been detected in human atheromatous plaque. Atherosclerosis involves an ongoing inflammatory response, reportedly involving chronic infection caused by multiple pathogens. The aim of this study was to examine the invasion of human aortic endothelial cells (HAECs) by oral viridans group streptococci and the subsequent cytokine production by viable invaded HAECs. The invasion of HAECs by bacteria was examined using antibiotic protection assays and was visualized by confocal scanning laser microscopy. The inhibitory effects of catalase and cytochalasin D on the invasion of HAECs were also examined. The production of cytokines by invaded or infected HAECs was determined using enzyme-linked immunosorbent assays, and a real-time polymerase chain reaction method was used to evaluate the expression of cytokine messenger RNA. The oral streptococci tested were capable of invading HAECs. The number of invasive bacteria increased with the length of the co-culture period. After a certain co-culture period, some organisms were cytotoxic to the HAECs. Catalase and cytochalasin D inhibited the invasion of HAECs by the organism. HAECs invaded by Streptococcus mutans Xc, Streptococcus gordonii DL1 (Challis), Streptococcus gordonii ATCC 10558 and Streptococcus salivarius ATCC 13419 produced more cytokine(s) (interleukin-6, interleukin-8, monocyte chemoattractant protein-1) than non-invaded HAECs. The HAECs invaded by S. mutans Xc produced the largest amounts of cytokines, and the messenger RNA expression of cytokines by invaded HAECs increased markedly compared with that by non-invaded HAECs. These results suggest that oral streptococci may participate in the pathogenesis of atherosclerosis.

We previously reported that detection of Streptococcus salivarius is feasible for proving the presence of saliva in a forensic sample. Here, a simple and rapid method for the detection of S. salivarius in forensic samples was developed that uses loop-mediated isothermal amplification (LAMP). The LAMP primer set was designed using S. salivarius-specific sequences of glucosyltransferase K. To simplify the procedure, the sample was prepared by boiling and mutanolysin treatment only, and the entire analytical process was completed within 2.5 h. The cut-off value was set at 0.1 absorbance units, measured at 660 nm, upon termination of the reaction. S. salivarius was identified in all saliva samples, but was not detected in other body fluids or on the skin surface. Using this method, S. salivarius was successfully detected in various mock forensic samples. We therefore suggest that this approach is useful for the identification of saliva in forensic practice.

We evaluated the antimicrobial activity of 25 monoketone curcuminoids (MKCs) against a representative panel of cariogenic bacteria in terms of their minimum inhibitory concentration (MIC) values. Curcumin A (10) displayed promising activity against Streptococcus mutans (MIC = 50 μg/ml) and Streptococcus mitis (MIC = 50 μg/ml) as well as moderate activity against S. sanguinis (MIC = 100 μg/ml), Lactobacillus casei (MIC = 100 μg/ml), and Streptococcus salivarius (MIC = 200 μg/ml). Results indicated higher activity of compound 10 than that of its bis-β-diketone analog. Additionally, compounds 3a (1,5-bis(4-methylphenyl)pentan-3-one) and 7b (1,5-bis(4-bromophenyl)pentan-3-ol) were moderately active against S. mitis (MIC = 100 μg/ml) and S. salivarus (MIC = 200 μg/ml).

Introduction: The effects of Streptococcus salivarius on the competence-stimulating peptide (CSP)-dependent biofilm formation by Streptococcus mutans were investigated. Methods: Biofilms were grown on 96-well microtiter plates coated with salivary components in tryptic soy broth without dextrose supplemented with 0.25% sucrose. Biofilm formations were stained using safranin and quantification of stained biofilms was performed by measuring absorbance at 492 nm. Results: S. mutans formed substantial biofilms, whereas biofilms of S. salivarius were formed poorly in the medium conditions used. Furthermore, in combination cultures, S. salivarius strongly inhibited biofilm formation when cultured with S. mutans. This inhibition occurred in the early phase of biofilm formation and was dependent on inactivation of the CSP of S. mutans, which is associated with competence, biofilm formation, and antimicrobial activity of the bacterium, and is induced by expression of the comC gene. Comparisons between the S. mutans clinical strains FSC-3 and FSC-3ΔglrA in separate dual-species cultures with S. salivarius indicated that the presence of the bacitracin transport ATP-binding protein gene glrA caused susceptibility to inhibition of S. mutans biofilm formation by S. salivarius, and was also associated with the regulation of CSP production by com gene-dependent quorum sensing systems. Conclusion: It is considered that regulation of CSP by glrA in S. mutans and CSP inactivation by S. salivarius are important functions for cell-to-cell communication between biofilm bacteria and oral streptococci such as S. salivarius. Our results provide useful information for understanding the ecosystem of oral streptococcal biofilms, as well as the competition between and coexistence of multiple species in the oral cavity.

Introduction: Microbial interactions are considered important in the adhesion process of pathogenic bacteria in the oral cavity. This study addressed the hypothesis that a streptococcal biofilm influences the hard tissue colonization by the periodontopathogen Aggregatibacter actinomycetemcomitans under hydrodynamic conditions. Methods: The colonization of a green-fluorescent-protein-labelled A. actinomycetemcomitans strain on surfaces coated with a streptococcal biofilm, was monitored in real time using a confocal laser scanning microscope-mounted flow cell. Culture and quantitative polymerase chain reaction data were obtained in parallel from a Modified Robbins Device. Results: Colonization of A. actinomycetemcomitans was inhibited by the four tested streptococci (Streptococcus sanguinis, Streptococcus cristatus, Streptococcus salivarius, and Streptococcus mitis). The most inhibiting species was S. sanguinis. Conclusion: These results confirmed the hypothesis that some bacterial species influence A. actinomycetemcomitans colonization of hard surfaces in vitro under hydrodynamic conditions.

Five yogurt batches were prepared with banana puree and sugar combinations and ABT-2 (Streptococcus salivarius ssp. thermophilus, Lactobaccilus acidophilus and Bifidobacterium spp.) as starter culture. The addition of fruit influenced (P

Survival and growth of Enterobacter sakazakii during processing and storage of plain yogurt were investigated. Preheated rehydrated milk was inoculated with a cocktail culture of E. sakazakii (103 cfu/mL of milk) and/or with thermophilic yogurt starter culture of Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus salivarius ssp. thermophilus. The inoculated milk was incubated at 40C for 5 h, then the samples were cooled and subsequently stored at 4C for up to 7 days. The results showed that E. sakazakii grew at an early stage of fermentation but declined at the end of the process. There was no significant difference between the populations of E. sakazakii in the presence or absence of lactic acid bacteria during the first 4 h of the incubation period but there was significant difference during the last hour of the incubation period. The populations of E. sakazakii decreased significantly during cooling and storage of yogurt (pH 4.2−4.7) compared with nonfermented milk samples at 4C. The presence of E. sakazakii did not have a significant effect on the growth of LAB during fermentation and storage of yogurt. The results obtained from this study indicate that the pH of yogurt and storage temperature were critical to the survival and growth of E. sakazakii in the manufacture of plain yogurt. PRACTICAL APPLICATIONS Enterobacter sakazakii prevalence in milk products and the production environment has been documented. The results obtained from this study may be of use to dairy producers to manufacture safe products using thermophilic lactic acid bacteria. These bacteria decreased the pH of milk in less than 5 h, resulting in E. sakazakii reduction. pH of yogurt during the fermentation process is a critical control point that should be monitored to produce safe products.

BACKGROUND: γ-Aminobutyric acid with several well-known physiological functions is biosynthesized via the irreversible α-decarboxylation of L-glutamate catalysed by glutamate decarboxylase (GAD). Although Streptococcus salivarius ssp. thermophilus has been widely applied to the dairy, the characterization of its GAD has not been reported. In this paper, the purification and the characterization of S. salivarius ssp. thermophilus GAD were investigated. RESULTS: GAD was purified 22-fold from crude protein extracts with a yield of 7.8% in five steps. The final preparation gave a single band on SDS-PAGE. The molecular weight of GAD determined by SDS-PAGE and gel filtration was 46.9 kDa and 103.6 kDa, respectively, indicating that the enzyme exists as a dimmer of homological subunits. The optimum temperature and pH of GAD was 55 °C and pH 4.0, respectively. The enzyme reacted only with L-glutamate among 19 α-amino acids with apparent Km at 2.3 mmol L−1 and did not react with D-glutamic acid. Activity of the enzyme could significantly be activated by 5 mmol L−1 of BaCl2 and inhibited by FeSO4, ZnSO4, CuSO4, MnSO4, Na2SO4, AgNO3, CoCl2, LiCl and KCl, respectively. The N-terminal amino acid sequence of GAD was NH2-MNEKLFREI. CONCLUSION: Both the characterization and the deduced amino sequence (ABI31651) showed the purified enzyme was a novel GAD. Copyright © 2008 Society of Chemical Industry

The culture conditions for glutamate decarboxylase (GAD) production under submerged fermentation by Streptococcus salivarius ssp. thermophilus were investigated. The results indicated the optimum culture medium was composed as follows: 15.0 g L−1 of peptone, 12.5 g L−1 of beef extract, 12.5 g L−1 of sucrose, 1.03 g L−1 of dipotassium hydrogen phosphate, 5 g L−1 of sodium acetate, 2 g L−1 of ammonium dibasic citrate, 2.12 g L−1 of calcium chloride, 1 g L−1 of Tween 80, and initial pH 6.79. The optimum culture temperature and time were 37 °C and 12 h, respectively. Under these conditions, GAD production was 257.46 ± 5.12 U, which was about 1.45-fold that of Man–Rogosa–Sharpe broth. Copyright © 2008 Society of Chemical Industry

Introduction: Periodontitis results from a shift in the subgingival micro. ora into a more pathogenic direction with Porphyromonas gingivalis, Prevotella intermedia, and Actinobacillus actinomycetemcomitans considered as periodontopathogens. In many cases, treatment procures only a temporary shift towards a less pathogenic microflora. An alternative treatment could be the deliberate colonization of pockets with antagonistic microorganisms to control the adhesion of periodontopathogens. The aim of this study was to identify bacterial strains that reduce adhesion of periodontopathogens to surfaces. Methods: Streptococcus sanguinis, Streptococcus crista, Streptococcus salivarius, Streptococcus mitis, Actinomyces naeslundii, and Haemophilus parainfluenzae were evaluated as potential antagonists against P. gingivalis ATCC 33277, P. intermedia ATCC 49046, and A. actinomycetemcomitans ATCC 43718 as periodontopathogens. Adhesion of periodontopathogens to the bottom plate of a parallel plate flow chamber was studied in the absence (control) and the presence of pre-adhering antagonistic strains up to a surface coverage of 5%. Results: The largest reduction caused by antagonistic strains was observed for P. gingivalis. All antagonistic strains except S. crista ATCC 49999 inhibited the adhesion of P. gingivalis by at least 1.6 cells per adhering antagonist, with the largest significant reduction observed for A. naeslundii ATCC 51655 (3.8 cells per adhering antagonist). Adhering antagonists had a minimal effect on the adhesion of A. actinomycetemcomitans ATCC 43718. Intermediate but significant reductions were perceived for P. intermedia, most notably caused by S. mitis BMS. Conclusion: The adhesion of P. gingivalis was inhibited best by antagonistic strains, while S. mitis BMS appeared to be the most successful antagonist.

Experimental conditions were established for the extraction, electrophoresis and detection of urease isoenzymes from Streptococcus salivarius. Thiol concentrations were critical and ureases from different strains varied in ease of dissociation. A characteristic pattern was obtained for 30 ureolytic S. salivarius strains isolated from; saliva (6), dental plaque (12), artificial dental plaque (6) and non-oral sources (6), and also for a ureolytic Streptococcus bovis from artificial plaque. One non-oral S. salivarius strain had ureases with slightly slower mobility. The electrophoretic pattern and mobility of ureases extracted from human mixed salivary bacteria were identical to those from S. salivarius except for an additional set of urease bands from unknown species of bacteria. There were no ureases from saliva matching those from Staphylococcus epidermidis--a contributor to ureolysis in artificial plaque. We conclude that there is considerable biochemical homogeneity among S. salivarius ureases and possibly other ureolytic streptococci. In saliva, urea is metabolized mainly by streptococcal ureases.

We show in this article that the transport of glucose, mannose and fructose by the phosphoenolpyruvate: mannose phosphotransferase system of oral streptococci requires the participation of a protein component that we have called IIIman. This protein was purified from Streptococcus salivarius by chromatography on DEAE-cellulose, DEAE-TSK, hydroxyapatite, and Dyematrex Green A. The purified protein migrated as a 38,900 molecular weight protein on a sodium dodecyl sulfate polyacrylamide gel. However, electrophoretic analysis of phosphoproteins and Western blot experiments indicated the presence in membrane-free cellular extracts of S. salivarius of 2 different forms of IIIman having molecular weights of 38,900 and 35,200. The presence of the high-molecular-weight form of IIIman was observed by immunodiffusion, Western blot and phosphorylation by [32]PEP in S. salivarius, Streptococcus mutans, Streptococcus sobrinus, and Streptococcus lactis but not in Streptococcus faecium, Staphylococcus aureus, Bacillus subtilis and Lactobacillus casei. Antibodies directed against the IIIman of S. salivarius did not react with the IIIman of Escherichia coli.

The antimicrobial properties of cements prepared from mechanically activated tetracalcium phosphate (maTTCP) were tested with the agar diffusion test using Streptococcus salivarius, Staphylococcus epidermidis, and a clinically isolated plaque mixture. All maTTCP cements showed a significantly higher antimicrobial potency as revealed by inhibition zones of ∼3–5 mm width, compared with a commercial Ca(OH)2//salicylate cement which only produced small inhibition zones around the cement specimens of 1.5 mm or less. This behavior was explained by the formation of amorphous Ca(OH)2 during setting of maTTCP cements, which is thought to have a higher solubility and may release more OH− ions than conventional Ca(OH)2//salicylate cements. In fact, the pH value in the agar gel around the specimens was higher in the case of maTTCP cements (7.8–8.7) compared with the Ca(OH)2//salicylate control (7.0–8.0). The maTTCP cements did not affect the photoactivation of resin-based composites, and their antimicrobial activity is making them interesting candidates for the use as pulp-capping agents, endodontic sealers, or cavity liners in dentistry. © 2007 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2007

Aim Recent UK National Institute for Health and Clinical Excellence guidelines state that there is no longer a need for oral antibiotic prophylaxis in patients undergoing dental procedures who are at risk of infective endocarditis (IE), and advocate the importance of maintaining good oral health. As viridans group streptococci (VGS) are common etiological agents of IE and inhabitants of the mouth, the purpose of this study was to examine the efficacy of common high-street mouthwashes against four classes of VGS organisms (salivarius, mitis, anginosus, and mutans groupings). Methods The survival of VGS, Streptococcus gordonii (National Collection of Type Cultures [NCTC] 7865), Streptococcus intermedius (NCTC 11324), Streptococcus mutans (NCTC 10449), Streptococcus oralis (NCTC 11427), Streptococcus pneumoniae (NCTC 7465, NCTC 7978, & American Type Culture Collection 49619) and Streptococcus salivarius (NCTC 8618) was assessed in vitro following treatment of approximately 107 c.f.u. in planktonic state with four mouthwashes. Results No organisms were culturable following 1-min exposure, and were not recovered following non-selective enrichment following incubation in Brain Heart Infusion broth supplemented with 0.8% (w/v) yeast extract. Conclusions These data indicate that such mouthwashes are able to completely kill VGS organisms tested in planktonic solution, where their use would promote good oral hygiene in patients at risk of IE.

It is difficult to distinguish mutans streptococci on the species level, and even more so on the subspecies level. Intact cell matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) (ICM) was applied to reference strains of five of the species of the mutans group (Streptococcus criceti, Streptococcus downei, Streptococcus mutans, Streptococcus ratti, Streptococcus sobrinus), nonmutans streptococci (Streptococcus oralis, Streptococcus mitis, Streptococcus salivarius, and Streptococcus sanguinis), and 177 mutans streptococci isolated from saliva of 10 children. From the analysis of the reference strains, readily distinguishable ICM mass spectra were obtained for the different species. Based on multivariate statistical analysis, a correct and unambiguous assignment was made of the spectra of 159 isolated mutans streptococci to S. mutans and 16 isolates to S. sobrinus. Two isolates were sorted out and were identified by sequencing of their 16S rRNA genes as Streptococcus anginosus. In addition, ICM indicated a misclassification for some reference strains (AHT, V 100 and E 49) and re-classified AHT and E 49 as S. ratti and V 100 as S. sobrinus. This was confirmed by 16S rDNA sequencing. Based on a statistical similarity analysis of the spectra of reference strains and a quantitative assessment of the reproducibility of ICM, the isolates identified as either S. mutans or S. sobrinus were phenotyped on the subspecies level. In the population of the clinical isolates, 14 unambiguously different S. mutans and three different S. sobrinus phenotypes were detected. ICM proved to be a powerful tool for a differentiation of mutans streptococci down to the subspecies level.

Zinc is a known inhibitor of acid production by mutans streptococci. Our primary objective was to extend current knowledge of the physiologic bases for this inhibition and also for zinc inhibition of alkali production by Streptococcus rattus FA-1 and Streptococcus salivarius ATCC 13419. Zinc at concentrations as low as 0.01-0.1 mm not only inhibited acid production by cells of Streptococcus mutans GS-5 in suspensions or in biofilms but also sensitized glycolysis by intact cells to acidification. Zinc reversibly inhibited the F-ATPase of permeabilized cells of S. mutans with a 50% inhibitory concentration of about 1 mm for cells in suspensions. Zinc reversibly inhibited the phosphoenolpyruvate: sugar phosphotransferase system with 50% inhibition at about 0.3 mm ZnSO4, or about half that concentration when the zinc-citrate chelate was used. The reversibility of these inhibitory actions of zinc correlates with findings that it is mainly bacteriostatic rather than bactericidal. Zinc inhibited alkali production from arginine or urea and was a potent enzyme inhibitor for arginine deiminase of S. rattus FA-1 and for urease of S. salivarius. In addition, zinc citrate at high levels of 10-20 mm was weakly bactericidal.

Twenty-three of 86 strains of lactic acid bacteria transformed linoleic acid into hydroxy FA. Two distinct conversion pathways were in operation. Two strains of Lactobacillus acidophilus and a strain of Pediococcus pentosaceus produced 13(S)-hydroxy-9-octadecenoic acid 113(S)-OH 18:11 and 10,13dihydroxyoctadecanoic acid (10,13-OH 18:0) as main and minor products, respectively, whereas 13 strains, including L. casei subsp. casei, L. paracasei subsp. paracasei, L. rhamnosus, L. lactis subsp. cremoris, and Streptococcus salivarius subsp. thermophilus produced 10-hydroxy-12-octadecenoic acid (10-OH 18:1). Seven strains of L. plantarum converted linoleic acid to 10-hydroxyoctadecanoic acid (10-OH 18:0) through 10-OH 18:1. Linoleic acid at 2 g/L was converted by L. acidophilus IFO13951T to 1.3 g of 13(S)-OH 18:1 and 0.09 g of 10,13-OH 18:0 in 7 d. Lactobacillus paracasei subsp. paracasei JCM 1111 produced 10-OH 18:1 in 91% yield, and L. plantarum JCM 8341, 10-OH 18:0 in 59% yield from linoleic acid (2 g/L) under optimal conditions. To our knowledge, this is the first report on the production of 13(S)-OH 18:1 by lactic acid bacteria other than ruminal bacteria, and of 10,13-OH 18:0 by any bacteria.

Streptococcus salivarius is divided into two serological subgroups that carry either fibrils or firnbriae. Although fimbriae have been observed on up to 50% of S. salivarius strains in the human oral cavity, no function has yet been assigned to them. To determine whether S. salivarius fimbriae have a role in adhesion, we examined the ability of S. salivarius to coaggregate with selected microorganisms involved in periodontal diseases. Our results show that S. salivarius coaggregated with Fusobacterium nucleatum, Porphyromonas gingivalis, and Prevotella intermedia. However, only fimbriated S. salivarius cells were able to coaggregate with P. intermedia, suggesting a specific role for these structures in the interaction. Heat treatment, sensitivity to sugars, amino acids, and EDTA, as well as protease treatment were also used to further characterize coaggregation between S. salivarius and periodontopathogens.

Thirty eight strains of oral streptococci were divided into six types by two-dimensional gel electrophoresis (2-DE) followed by glycosyltransferase (GLT) activity stain: Type 1, Streptococcus mutans; Type 2, Streptococcus rattus; Type 3, Streptococcus sobrinus and Streptococcus downei; Type 4, Streptococcus cricetus; Type 5, Streptococcus salivarius; and Type 6, Streptococcus sanguis, Streptococcus oralis and Streptococcus gordonii. In Types 1, 2 and 5, two glucosyltransferases synthesizing water-insoluble (GTF-I) and water-soluble glucans (GTF-S) and a fructosyltransferase (FTF) were detected. In Types 3 and 4, GTF-I and two GTF-Ss were detected. Only one GTF-S was detected in Type 6. The 2-DE patterns for these six types were clearly distinguished from each other based on the kind, number and location of GLTs in gel.

Background and Objectives One of the main advantages of laser surgery is it's bactericidal effect which reduces the risk of postoperative infections. Several study designs have been set to investigate this effect. Aim of this study was to research if the bactericidal effect of laser tool was affected from several factors in vitro studies. Study Design/Materials and Methods To determinate and investigate the bactericidal effect of laser in an original model, α-hemolytic streptococcus, Bacterioides fragilis, Neisseria, Streptococcus salivarius, Staphylococcus aureus, and Candida albicans were prepared in 104, 106 and 108 inoculum and placed in Mueller-Hinton Broth which have five different proportions of sheep blood. Samples which exposed with various energy levels of Nd:YAG laser were spread on agar plates, and at the end of an incubation time the colonization counted comparatively. The lowest energy level without colonization was accepted as minimal bactericidal energy level. Results Highest minimum bactericidal energy level is used for α-hemolytic streptococcus and lowest values for neisseria. Bactericidal effect decreased on suspensions, of which population of microorganisms are high and hemoglobin concentration was high in the broth. Conclusions These findings suggest that the Nd:YAG laser has a higher bactericidal effect when sheep blood is added to the media. Factors like population and type of bacteria in the irradiated suspension affect minimum bactericidal energy level. Lasers Surg. Med. 32:197–202, 2003. © 2003 Wiley-Liss, Inc.

A pH-sensitive fluorescent dye, 2', 7'-bis-(2-carboxyethyl)-5 and 6-carboxyfluorescein (BCECF), was used to determine intracellular pH (pH(in)). The efflux of BCECF loaded into oral streptococcal cells was determined after incubation of the cells at 35 degrees C for 20 min in the presence and absence of glucose. In the absence of glucose, the fluorescence of intracellular BCECF in Streptococcus mutans, Streptococcus sanguis, Streptococcus salivarius and Streptococcus sobrinus decreased only very slightly, indicating that the dye could be useful for pH(in) determination. In the presence of glucose, however, the fluorescence decreased by 57%. Thus, the pH(in) of S. mutans cells was measured by the BCECF method in the absence of glucose at various acidic pH levels by adding lactic, acetic and hydrochloric acids to the cell suspensions. The pH(in) was almost equal to the extracellular pH (pH(out)) for pH(out) values of between 8 and 5, indicating that protons permeated easily across the S. mutans cell membrane. For pH(out) between 5 and 4, pH(in) was constant at around 5, suggesting that the cell membrane was impermeable to protons, or that a cytoplasmic buffering system functioned. pH(in) decreased at pH(out) values of < 4. The constant pH(in) at acidic pH(out) levels could protect intracellular components, such as proteins, against acidification by sugar fermentation.

Dextranase (Dex) is an enzyme that hydrolyzes glucan, a polymer of glucose synthesized from sucrose by glucosyltransferases (GTFs). By comparing amino acid sequences of Dexs and GTFs, we found that the Dex enzymes of Streptococcus mutans, Streptococcus sobrinus, Streptococcus downei and Streptococcus salivarius had similar amino acid sequences to those of the catalytic sites of GTFs of mutans streptococci. We therefore examined the amino acid essential in Dex catalysis by molecular genetic approaches in this study. Site-directed mutagenesis was used to convert the Asp-385 of the Dex molecule of S. mutans Ingbritt to Glu, Asn, Thr or Val. Replacement of Asp-385 with any of the amino acids resulted in complete disappearance of Dex activity. However, replacement of other Asp residues did not affect the enzyme activity. The inactive enzymes still retained dextran-binding ability. These results suggest that Asp-385 of the Dex of S. mutans Ingbritt was essential for enzyme activity and the catalytic and substrate-binding sites were located at different sites within the Dex molecule.

Objectives:Nasopharyngeal colonization is the initial step in pathogenesis of pneumococcal diseases, including otitis media. Previous studies suggested that Streptococcus salivarius, a commercial probiotic, could be a potential pharyngeal probiotic. We investigated the effects of S salivarius on pneumococcal colonization and otitis media by in vitro and in vivo models of infection and colonization pattern of S salivarius by pilot study.Methods:Descriptive study conducted at Melbourne University and Murdoch Childrens Research Institute, Melbourne from 2011 to 2013. High, medium, or low numbers of S salivarius were added before, with, or after pneumococcal administration to human epithelial (CCL-23) cells in vitro. The percent colonization of pneumococci was determined after 3 hours of incubation. In experiments using infant mice, repeated pernasal administration of S salivarius (~107 CFU/dose) was used as an intervention in pneumococcal colonization model and otitis media. Colonization of upper respiratory...

OBJECTIVE To compare the microbial flora of adenoids from patients with recurrent acute otitis media (AOM) and patients with obstructive sleep apnea (OSA). MATERIALS AND METHODS This study was prospective and controlled. Adenoids were obtained from children undergoing adenoidectomy for recurrent AOM (n=7) or OSA (n=13). Specimens were processed for total deoxyribonucleic acid (DNA) isolation. 16s DNA 454-pyrosequencing was performed on AOM (n=5) and OSA (n=5) specimens. All specimens were analyzed by real-time polymerase chain reaction for the quantification of the oral commensal bacteria, Streptococcus salivarius. RESULTS All adenoid specimens had evidence of microbes. Haemophilus influenzae, Moraxella catarrhalis, Streptococcus pneumoniae, Pseudomonas aeruginosa, and Staphylococcus aureus were among the dominant species in all samples. S. salivarius, Prevotella sp. and Terrahaemophilus aromaticivorans were more common on adenoids from OSA patients (p

A surface protein antigen (PAc) of Streptococcus mutans, particularly the A-region of the molecule, has been reported to interact with salivary components on the tooth surface. It might be a candidate antigen inducing the production of antibodies against the adherence of S. mutans to the tooth surface. We investigated the effects of monoclonal antibodies (MoAbs) obtained by immunization of synthetic PAc peptides that completely correspond to the amino acid sequence of part of the A-region. These MoAbs recognize several core B-cell epitopes in the sequence. Two (KH5 and SH2) of these antibodies reacted with both S. mutans and Streptococcus sobrinus, but not with Streptococcus sanguis, Streptococcus salivarius, Porphyromonas gingivalis or Lactobacillus casei. They clearly inhibited the real-time adherence of S. mutans to salivary components in a biosensor. KH5, which showed a real-time inhibition (71%), also significantly prevented the recolonization of S. mutans on the tooth surface in rats. These results suggested that the core B-cell epitope (-Y---L--Y----) recognized by KH5 was the essential sequence in the antigenic epitopes of PAc protein recognized specifically by the inhibitory antibody. Therefore, the amino acid residues were found to be important in the initial attachment of S. mutans to the tooth surface. These results provide for the mechanism of PAc molecule in the initial attachment of S. mutans on the tooth surface and more effective designs for the removal of S. mutans and S. sobrinus from the oral cavity.

Background: Acetaldehyde is a known local carcinogen in the digestive tract in humans. Bacterial overgrowth in the hypochlorhydric stomach enhances production of acetaldehyde from ethanol in vivo after alcohol ingestion. Therefore, microbially produced acetaldehyde may be a potential risk factor for alcohol-related gastric and cardiac cancers. This study was aimed to investigate which bacterial species and/or groups are responsible for acetaldehyde formation in the hypochlorhydric human stomach and to characterize their alcohol dehydrogenase (ADH) enzymes. Methods: After 7 days of treatment with 30 mg of lansoprazole twice a day, a gastroscopy was performed on eight volunteers to obtain hypochlorhydric gastric juice. Samples were cultured and bacteria were isolated and identified; thereafter, their acetaldehyde production capacity was measured gas chromatographically by incubating intact bacterial suspensions with ethanol at 37°C. Cytosolic ADH activities, Km values, and protein concentration were determined spectrophotometrically. Results: Acetaldehyde production of the isolated bacterial strains (n= 51) varied from less than 1 to 13,690 nmol of acetaldehyde/109 colony-forming units/hr. ADH activity of the strains that produced more than 100 nmol of acetaldehyde/109 colony-forming units/hr (n= 23) varied from 3.9 to 1253 nmol of nicotinamide adenine dinucleotide per minute per milligram of protein, and Km values for ethanol ranged from 0.65 to 116 mM and from 0.5 to 3.1 M (high Km). There was a statistically significant correlation (r= 0.64, p < 0.001) between ADH activity and acetaldehyde production from ethanol in the tested strains. The most potent acetaldehyde producers were Neisseria and Rothia species and Streptococcus salivarius, whereas nearly all Stomatococcus, Staphylococcus, and other Streptococcus species had a very low capacity to produce acetaldehyde. Conclusions: This study demonstrated that certain bacterial species or groups that originate from the oral cavity are responsible for the bulk of acetaldehyde production in the hypochlorhydric stomach. These findings provide new information with the respect to the local production of carcinogenic acetaldehyde in the upper digestive tract of achlorhydric human subjects.

Urease synthesis in Streptococcus salivarius is induced by an acid environment, carbohydrate and a high growth rate. We now report that both cysteine and sulfide above 1 mM strongly suppress S. salivarius urease levels. Close structural relatives of cysteine (cysteamine, ethanedithiol and penicillamine) at 5 mM buffered to pH 7.0 also caused urease suppression, but thiols in general (2-mercaptoethanol, dithiothreitol and glutathione) did not. In cultures buffered below pH 5.9, the cysteine-induced urease suppression was lifted substantially, but the sulfide suppression increased, suggesting involvement of different processes. Urease activity was inhibited 50% by 5 mM mercaptoethanol but unaffected by 5 mM cysteine or sulfide, hence modification of enzyme activity by thiols is not directly related to suppression of their levels in culture. Cysteine, arising primarily through protein hydrolysis which also raises the pH, could be a surrogate pH feedback signal for nearby alkaline conditions, and sulfide may reflect activity of periodontopathic plaques.

This study investigated the oral microbiota of children undergoing liver transplantation. Oral swabs were taken using a standardized procedure from 27 children before liver transplantation and at 3 and 100 days post-transplantation and from 27 healthy controls at baseline and 90 days. Viridans streptococci, yeasts, staphylococci, enterococci and Enterobacteriaceae were enumerated and identified using conventional techniques. The oral microbiota of the patients changed significantly immediately post-transplantation, but by the final examination, it had returned to baseline levels. The oral microbiota of the controls did not change significantly. The numbers and proportions of Streptococcus salivarius, Streptococcus sanguis and Streptococcus gordonii as percentages of the total streptococcal counts and of the total anaerobic counts decreased significantly 3 days post-transplantation (P < or = 0.006). There were no significant changes in the numbers and proportions of Streptococcus oralis and Streptococcus mitis isolated pre- and post-transplantation. The isolation frequencies and numbers of yeasts were significantly higher in patients than controls. Staphylococci were isolated in low numbers from all children. Enterococci and Enterobacteriaceae were isolated infrequently from patients.

The metabolism of urea by urease enzymes of oral bacteria profoundly influences oral biofilm pH homeostasis and oral microbial ecology. The purpose of this study was to gain insight into the regulation of expression of the low pH-inducible urease genes in populations of Streptococcus salivarius growing in vitro in biofilms and to explore whether urease regulation or the levels of urease expression in biofilm cells differed significantly from planktonic cells. Two strains of S. salivarius harbouring urease promoter fusions to a chloramphenicol acetyltransferase (cat) gene were used: PurelCAT, containing a fusion to the full-length, pH-sensitive promoter; or Pureldelta100CAT, a constitutively derepressed deletion derivative of the urease gene promoter. The strains were grown in a Rototorque biofilm reactor in a tryptone-yeast extract-sucrose medium with or without pH control. Both CAT and urease activities in biofilms were measured at 'quasi-steady state' and after a 25mM glucose pulse. The results showed that CAT expression in PurelCAT was repressed at relatively neutral pH values, and that expression could be induced by acidic pH after carbohydrate challenge. Biofilms of PurelCAT grown at low pH, without buffering, had about 20-fold higher CAT levels, and only a modest further induction could be elicited with carbohydrate pulsing. The levels of CAT in biofilms of PurelCAT grown in buffered medium were slightly higher than those reported for planktonic cells cultured at pH 7.0, and the levels of CAT in Purel-CAT growing at low pH or after induction were similar to those reported for fully induced planktonic cells. CAT activity in Pureldelta100CAT was constitutively high, regardless of growth conditions. Interestingly, urease activity detected in biofilms of the parent strain, S. salivarius 57.1, could be as much as 130-fold higher than that reported for fluid chemostat cultures grown under similar conditions. The higher level of urease activity in biofilms was probably caused by the accumulation of the stable urease enzyme within biofilm cells, low pH microenvironments and the growth phase of populations of cells in the biofilm. The ability of S. salivarius biofilm cells to upregulate urease expression in response to pH gradients and to accumulate greater quantities of urease enzyme when growing in biofilms may have a significant impact on oral biofilm pH homeostasis and microbial ecology in vivo. Additionally, S. salivarius carrying the pH-sensitive urease gene promoter fused to an appropriate reporter gene may be a useful biological probe for sensing biofilm pH in situ.

Parabens were found to be potent inhibitors of alkali production from arginine by oral streptococci such as Streptococcus rattus, Streptococcus sanguis and Streptococcus gordonii. For example, 2 μmol butylparaben per ml completely and irreversibly inhibited arginolysis by intact cells of S. rattus FA-1 and was lethal for the organism. In contrast, butylparaben was not a very effective inhibitor of ureolysis by intact cells of Streptococcus salivarius 57.I, although it did kill the cells. Butylparaben irreversibly inhibited the cytoplasmic enzymes arginine deiminase, carbamate kinase and urease in permeabilized cells or isolated form. However, inhibition of arginolysis by intact cells appeared to be due primarily to irreversible inhibition of transport systems for arginine uptake, because butylparaben added to intact cells did not reduce levels of arginine deiminase when the cells were subsequently permeabilized after washing. The insensitivity of ureolysis by intact cells to butylparaben can be related to the known high permeability of cell membranes to urea and the cytoplasmic location of urease. The potency of butylparaben as an inhibitior of arginolysis or glycolysis and as a lethal agent was found to be greater at acid pH that at neutral or alkaline pH.

Background : Although 5-aminosalicylic acid (5-ASA) oral compounds are the standard maintenance treatment for ulcerative colitis in remission, some patients cannot use them because of side-effects. Clinical and experimental observations have suggested the potential role of probiotics in inflammatory bowel disease therapy. Aim : To evaluate the effects on intestinal microflora and the clinical efficacy of a new probiotic preparation in patients with ulcerative colitis in remission. Patients and methods : Twenty patients with ulcerative colitis, intolerant or allergic to 5-ASA, have been treated with a new probiotic preparation (VSL # 3, CSL, Milan, Italy) containing 5 × 1011 cells/g of 3 strains of bifidobacteria, 4 strains of lactobacilli and 1 strain of Streptococcus salivarius ssp. thermophilus. Two doses of 3 g were administered o.d. for 12 months. Faecal samples for stool culture were obtained from the patients at the beginning of the trial and after 10, 20, 40, 60, 75, 90 days, 12 months and at 15 days after the end of the treatment. The following bacterial groups have been evaluated in the faeces: total aerobic and anaerobic bacteria, enterococci, Streptococcus thermophilus, lactobacilli, bifidobacteria, Bacteroides, clostridia, coliforms. Patients were assessed clinically every two months, and assessed endoscopically at 6 and 12 months or in relapse. Results : Faecal concentrations of Streptococcus salivarius ssp. thermophilus, lactobacilli and bifidobacteria increased significantly in all patients, compared to their basal level, from the 20th day of treatment (P

Adamo HL, Buruiana R, Schertzer L, Boylan RJ. A comparison of MTA, Super-EBA, composite and amalgam as root-end filling materials using a bacterial microleakage model. International Endodontic Journal, 32, 197‐203, 1999. Aim The aim of this study was to compare traditional and newly developed root-end filling materials for resistance to bacterial microleakage. Methodology Sixty extracted single-rooted teeth were randomly divided into five groups for root-end filling with mineral trioxide aggregate, Super-EBA, TPH composite resin with ProBond dentine bonding agent, Dispersalloy amalgam with and without ProBond, and positive and negative control groups. Root canals were instrumented using the step-back technique and simulated root-end resections performed. Root-end filling materials were placed in 3 mm ultrasonic retropreparations. Nail varnish was applied to all external root surfaces to the level of the resected root ends to prevent lateral microleakage. Samples were sterilized in an ethylene oxide sterilizer for 12 h. Using a newly designed model system, the apical 3‐4 mm of the roots were immersed in BHI culture medium with phenol red indicator within culture chambers. The coronal access of each specimen was inoculated every 48 h with a suspension of Streptococcus salivarius. Culture media were observed every 24 h for colour change indicating bacterial contamination. Media demonstrating colour change were plated for S. salivarius. Samples were observed for 12 weeks. Results At 4 weeks 10% of specimens from each experimental group had evidence of leakage. At 8 weeks 20% of specimens filled with amalgam without dentine bonding agent, Super-EBA and MTA had evidence of leakage. At 12 weeks minor differences between materials were noted. Conclusions Under the conditions of the study, despite some variations, there were no statistically significant differences in rate of microleakage among the five groups tested at either 4, 8 or 12 weeks.

Aim This study was undertaken to determine the antimicrobial effect of various endodontic irrigants against six selected microorganisms MethodologyStaphylococcus aureus, Enterococcus faecalis, Streptococcus salivarius, Str. pyogenes, Escherichia coli and Candida albicans were included in the study. Pre-sterilized Whatman paper discs, 6 mm in diameter and soaked with the test solution, were prepared and placed onto the previously seeded agar Petri plates. Each plate was incubated aerobically. A zone of inhibition was recorded for each plate and the results were analysed statistically. Results 5.25% NaOCl was effective against all test microorganisms with a substantial zone of inhibition. Saline was always ineffective. Decreased concentration of NaOCl significantly reduced its antimicrobial effect. Cresophene showed a significantly larger (P > 0.05) average zone of inhibition compared to the other experimental irrigants. Alcohol had smaller but not significantly different zones of inhibition than chlorhexidine. Conclusions 5.25% NaOCl was superior in its antimicrobial abilities compared with other irrigants used. A reduced concentration of NaOCl (0.5%) resulted in significantly decreased antimicrobial effects. When compared with 21% alcohol, 0.5% NaOCl and 2% chlorhexidine, paramonochlorophenol (cresophene) showed a greater antimicrobial effect.