Association of free amino acids with caries experience and mutans streptococci levels in whole saliva of children with early childhood caries

Objective: The aim of the present study was to identify the free amino acid content in whole saliva of children with (CE) and without early childhood caries (CF) (ECC), correlating these findings with caries experience and mutans streptococci (MS) levels in saliva.Design: Seventy-eight healthy children, both genders, 6-71 months age, were selected to participate in the study. Following examination for dmft scores calculation, unstimulated whole saliva was collected from all 78 participants, stored at -80 degrees C, and used for amino acid analysis, on a Biochem 20 plus amino acid analyzer. Stimulated whole saliva was collected from 52 children, transported, diluted and plated on MSB agar medium for detection of MS in cfu/mL.Results: Forty different free amino acids were identified in whole saliva, with great variation in their concentration. A statistically significant relation was found between caries experience and the presence of free proline and glycine. While proline (p = 0.0182) was more frequently absent in the CF group, the absence of glycine (p = 0.0397) was more often observed in the CE group. In the presence of higher levels of MS, free glycine reduced the risk of experiencing dental caries (p = 0.0419). Conversely, the presence of proline was found to increase the risk of experiencing the disease (p = 0.0492).Conclusions: The presence of free proline and absence of free glycine in children with ECC, highly contaminated with MS, increased the chances of experiencing dental caries in the present population. Further studies are needed to better understand this phenomenon. (C) 2008 Elsevier Ltd. All rights reserved.

Estreptomicina aumenta a aderência em células epiteliais de Bacteroides melaninogenicus associado às lesões peridentárias da cara inchada dos bovinos

Cara inchada of cattle (CI) is a periodontitis which occurs in calves in several regions of Brazil. Its progressive lesions, caused mainly by Bacteroides melaninogenicus and Actinomyces pyogenes, could lead to loss of teeth, malnutrition and possibly death of affected animals. For further analysis of the pathogenesis of this disease the influence of streptomycin on the adherence of the causative bacteria to oral epithelial cells of cattle was determined. Production of streptomycin by soil bacteria (Actinomycetes) in Brazil had been suggested by several authors. In our studies, streptomycin in subinibitory concentrations increased tenfold the rate of adherence with B. melaninogenicus (from 0.4 to 4.0 bacteria per bovine epithelial cell). On the contrary, streptomycin did not influence significantly the adherence of A. pyogenes nor streptococci of the viridans group. Thus, streptomycin could be a contributing factor to the development of the periodontal infections with B. melaninogenicus.

Correlation among mutans streptococci counts, dental caries, and IgA to Streptococcus mutans in saliva

Two-hundred and forty individuals were studied, divided into five groups as follows: caries-free children, children with caries, children with rampant caries, young adults with and without caries. Whole stimulated saliva was collected and all individuals were investigated for DMFT/dmft according to the WHO criteria and the simplified oral hygiene index (OHI-S). Quantitative analysis of the total aerobic flora and mutans streptococci in saliva was performed. Also, the level of salivary anti-S. mutans IgA was determined by ELISA. Children with rampant caries showed the highest OHI-S value. The highest total counts of microorganisms were found in the group of children with caries. No statistically significant differences were observed for salivary flow, OHI-S and microorganism counts between the groups of young adults. No correlation between mutans streptococci counts and anti-Streptococcus mutans IgA levels was observed in the studied groups. A correlation between increased anti-Streptococcus mutans IgA levels and caries-free status was observed among young adults but not among children.

Arbitrarily primed polymerase chain reaction for fingerprinting the genotype identification of mutans streptococci in children with Down syndrome

This study investigated the possible intrafamilial similarity of mutans streptococcal strains in some families with a child with Down syndrome using chromosomal DNA fingerprinting. The isolates were genotyped using arbitrarily primed polymerase chain reaction with the OPA 02 and OPA 03 primers. The results showed that five children with Down syndrome harbored mutans streptococci genotypes different from those of their mothers. A matching of genotypes was observed within the control pair (mother/child without Down syndrome). After six months, new samples were collected from all participants. Analysis showed that samples from children with Down syndrome were colonized by a new strain of Streptococcus mutans that did not match the previously collected one. The results suggest the S. mutans indigenous bacteria change more than once in children with Down syndrome.

Effect of 0.4% stannous fluoride gel on Streptococci mutans in relation to elastomeric rings and steel ligatures in orthodontic patients

Background: Patients with fixed orthodontic appliances often experience an absolute increase in the number of Streptococci mutans colony-forming units (cfu). The aim of this investigation was to study the development of biofilm and S. mutans cfu in connection with stainless steel ligatures and elastomeric rings in orthodontic patients treated with and without 0.4% stannous fluoride gel (SFG). Material: Forty-seven patients were divided into 2 groups: those treated with 0.4% SFG for 4 minutes (experimental) and those without 0.4% SFG (control). In each patient, elastomeric rings were used for ligation on 1 side of the dental arch midline, and stainless steel ligatures were used on the opposite side. Saliva samples were collected before and after appliance placement. At 15 and 30 days after appliance placement, biofilm samples from the stainless steel ligatures and the elastomeric rings were collected and subjected to microbiologic procedures and scanning electron microscopy (SEM) analysis. Results: The numbers of S. mutans cfu in the saliva and biofilm were not statistically different between the teeth fitted with elastomeric rings and stainless steel ligatures, or between the experimental and control groups. SEM analysis showed biofilm formation on both ligature ties. Conclusions: Topical application of 0.4% SFG in orthodontic patients with elastomeric rings or stainless steel ligatures does not cause a significant decrease in S. mutans cfu in the saliva and biofilm. Copyright © 2005 by the American Association of Orthodontists.

Elimination of intracanal infection in dogs' teeth with induced periapical lesions after rotary instrumentation: Influence of different calcium hydroxide pastes

The aim of this study was to evaluate the antiseptic efficacy of rotary instrumentation associated with calcium hydroxide-based pastes prepared with different vehicles and antiseptics. Chronic periapical lesions were experimentally induced in 72 premolar root canals of four dogs. Under controlled asepsis, after initial microbiological sampling (A1), the root canals were instrumented using the ProFile system in conjunction with 5.25% sodium hypochlorite and the intracanal medication was placed. Four experimental groups were formed according to the pastes used: group 1- Calen (n=18), group 2- Calen+CPMC (n=20), group 3- Ca(OH)2 p.a.+ anaesthetic solution (n=16) and group 4- Ca(OH)2 p.a.+ 2% chlorhexidine digluconate (n=18). After 21 days, the pastes were removed; the canals were emptied and 96 hours later a second microbiological sample was obtained (A2). The incidence of positive microbiological cultures and the number of cfus in stages A1 and A2 were compared statistically by the Wilcoxon test while the influence of the different treatments in intracanal infection was evaluated by Kruskal-Wallis test at 5% significance level (p<0.05). Large numbers of strict and facultative anaerobes, and viridans group streptococci were found in 100% of root canals of A1 samples. Among A2 samples, all treatments showed significant reduction of cfus and positive cultures (p<0.05), but only groups 3 and 4 showed 100% of root canals free of microorganisms. Rotary instrumentation plus NaOCl 5.25% associated with intracanal medication produced a drastic reduction or elimination of intracanal microbiota, whose performance was not influenced by the nature of the vehicle or the antiseptic added to the Ca(OH)2 p.a.

Influence of preventive dental treatment on mutans streptococci counts in patients undergoing head and neck radiotherapy

The aim of this study was to evaluate the influence of chlorhexidine gluconate, sodium fluoride and sodium iodine on mutans streptococci counts in saliva of irradiated patients. Forty-five patients were separated into three experimental groups and received chlorhexidine (0.12%), sodium fluoride (0.5%) or sodium iodine (2%), which were used daily during radiotherapy and for 6 months after the conclusion of the treatment. In addition, a fourth group, composed by 15 additional oncologic patients, who did not receive the mouthwash or initial dental treatment, constituted the control group. Clinical evaluations were performed in the first visit to dental clinic, after initial dental treatment, immediately before radiotherapy, after radiotherapy and 30, 60, 90 days and 6 months after the conclusion of radiotherapy. After clinical examinations, samples of saliva were inoculated on SB20 selective agar and incubated under anaerobiosis, at 37oC for 48 h. Total mutans streptococci counts were also evaluated by using real-time PCR, through TaqMan system, with specific primers and probes for S. mutans and S. sobrinus. All preventive protocols were able to reduce significantly mutans streptococci counts, but chlorhexidine gluconate was the most effective, and induced a significant amelioration of radiotherapy side effects, such as mucositis and candidosis. These results highlights the importance of the initial dental treatment for patients who will be subjected to radiotherapy for head and neck cancer treatment.

Recovery of mutans streptococci on MSB, SB-20 and SB-20M agar media

Objective: The recovery of mutans streptococci in saliva and dental biofilm samples depends, in part, on the culture medium used. In this study, we compared (i) the culture media Sucrose-Bacitracin agar (SB-20), Modified SB-20 (SB-20M) and Mitis Salivarius Bacitracin agar (MSB) in the count of colony forming units (cfu) of mutans streptococci and (ii) in the morphological and biochemical differentiation between Streptococcus mutans and Streptococcus sobrinus. Design: Samples of non-stimulated saliva from 20 children were plated on SB-20, SB-20M and MSB, and incubated in microaerophilia at 37 °C for 72 h. Identification of microorganisms was based on analysis of colony morphology under stereomicroscopy. The biochemical identification of colonies was done by biochemical tests using sugar fermentation, resistance to bacitracin and hydrogen peroxide production. Results: There was no significant difference (p > 0.05) in the number of cfu of mutans streptococci recovered on SB-20 and SB-20M agar. Comparing the media, SB-20 and SB-20M yielded a larger number of mutans streptococci colonies (p < 0.05) and were more effective than MSB in the identification of S. sobrinus (p < 0.05), but not of S. mutans (p > 0.05). Conclusion: There was no significant difference between SB-20 and SB-20M culture media in the count of mutans streptococci, demonstrating that the replacement of sucrose by coarse granular cane sugar did not alter the efficacy of the medium. Compared with MSB, SB-20 and SB-20M allowed counting a larger number of mutans streptococci colonies and a more effective morphological identification of S. sobrinus. © 2012 Elsevier Ltd.

Antimicrobial activity of the synthetic peptide Lys-a1 against oral streptococci

The peptide LYS-[TRP6]-Hy-A1 (Lys-a1) is a synthetic derivative of the peptide Hy-A1, initially isolated from the frog species Hypsiboas albopunctatus. According to previous research, it is a molecule with broad antimicrobial activity. The objective of this study was to evaluate the antimicrobial activity of the synthetic peptide Lys-a1 (KIFGAIWPLALGALKNLIK- NH2) on the planktonic and biofilm growth of oral bacteria. The methods used to evaluate antimicrobial activity include the following: determination of the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) in microtiter plates for growth in suspension and quantification of biomass by crystal violet staining and counting of colony forming units for biofilm growth. The microorganisms Streptococcus oralis, Streptococcus sanguinis, Streptococcus parasanguinis, Streptococcus salivarius, Streptococcus mutans and Streptococcus sobrinus were grown in Brain Heart Infusion broth at 37 °C under atmospheric pressure with 10% CO2. The peptide was solubilized in 0.1% acetic acid (v/v) at various concentrations (500-1.9 μg mL-1). Chlorhexidine gluconate 0.12% was used as the positive control, and BHI culture medium was used as the negative control. The tested peptide demonstrated a remarkable antimicrobial effect, inhibiting the planktonic and biofilm growth of all strains tested, even at low concentrations. Thus, the peptide Lys-a1 is an important source for potential antimicrobial agents, especially for the control and prevention of microbial biofilms, which is one of the most important factors in cariogenic processes. © 2012 Elsevier Inc.

The effect of chlorhexidine on plaque index and mutans streptococci in orthodontic patients: A pilot study

Aim: The purpose of this study was to assess chlorhexidine effects on plaque index and salivary levels of mutans streptococci (MS) when used as the immersion solution for removable orthodontic appliances and added to their acrylic resin composition. Methods: Forty-five patients (6 to 12 years old) were randomly assigned into three groups with 15 patients each. Group I (control)—without orthodontic appliances disinfection; Group II—removable orthodontic appliances which had been immersed in 0.12% chlorhexidine digluconate overnight (8 hours), and Group III—orthodontic appliances in which 0.12% chlorhexidine digluconate solution had been incorporated into their resin composition. Saliva was collected for quantification of MS and evaluation of plaque index was performed before and after installation of orthodontic appliance at 0, 2, 4, 6, 8, and 10 weeks. Data were analyzed by using analysis of variance. Results: Number of MS colonies in saliva and plaque index showed no statistically differences among groups at the different periods (p > 0.05). Conclusions: It could be concluded that chlorhexidine incorporation into the acrylic resin of removable orthodontic appliances at 0.12% concentration and immersion of the appliance into 0.12% chlorhexidine solution were not effective in reducing plaque index and the number of MS in saliva.

Are mutans streptococci detected in preschool children a reliable predictive factor for dental caries risk? A systematic review

Research suggests that mutans streptococci play an important role in cariogenesis in children but the usefulness of bacterial testing in risk assessment is unknown. Our objective was to summarize the literature assessing the association of mutans streptococci and dental caries in preschool children, (Pre)Medline (1966-2003), Embase (1980-2003), the Cochrane Register of Controlled Trials (2003, issue 3), and reference lists of included studies were searched. All abstracts found by the electronic searches (n = 981) were independently scrutinized by 2 reviewers. Minimal requirements for inclusion were assessment of preschool children without caries at baseline, reporting of mutans streptococci present in saliva or plaque at baseline and assessment of caries presence after a minimum of 6 months of follow-up. Participants' details, test methods, methodological characteristics and findings were extracted by one reviewer and cross-checked by another. Homogeneity was tested using chi2 tests. Results of plaque and saliva testing were pooled separately using a fixed effects model. Methodological quality of reports was low. Out of 9 studies included, data from 3 reports on plaque test assessment alone (n = 300) and from 4 reports on saliva test assessment alone (n = 451) were available for pooled analysis. The pooled risk ratio (95% CI) was 3.85 (2.48-5.96) in studies using plaque tests and 2.11 (1.47-3.02) in those using saliva testing. Presence of mutans streptococci, both in plaque or saliva of young caries-free children, appears to be associated with a considerable increase in caries risk. Lack of adjustment for potential confounders in the original studies, however, limits the extent to which interpretations for practice can be made.

Brain injury in experimental neonatal meningitis due to group B streptococci

We have characterized the pattern of brain injury in a rat model of meningitis caused by group B streptococci (GBS). Infant rats (12-14 days old; n = 69) were infected intracisternally with 10 microliters of GBS (log10(2.3) to 4.5 colony-forming units). Twenty hours later, illness was assessed clinically and cerebrospinal fluid was cultured. Animals were either immediately euthanized for brain histopathology or treated with antibiotics and examined later. Early GBS meningitis was characterized clinically by severe obtundation and seizures, and histopathologically by acute inflammation in the subarachnoid space and ventricles, a vasculopathy characterized by vascular engorgement, and neuronal injury that was most prominent in the cortex and often followed a vascular pattern. Incidence of seizures, vasculopathy and neuronal injury correlated with the inoculum size (p < 0.01). Early injury was almost completely prevented by treatment with dexamethasone. Within days after meningitis, injured areas became well demarcated and showed new cellular infiltrates. Thirty days post-infection, brain weights of infected animals treated with antibiotics were decreased compared to uninfected controls (1.39 +/- 0.18 vs 1.64 +/- 0.1 g; p < 0.05). Thus, GBS meningitis in this model caused extensive cortical neuronal injury resembling severe neonatal meningitis in humans.

[Endocarditis prophylaxis with amoxicillin, clindamycin or erythromycin? Serum bacteriostatic and bactericidal effect against Streptococcus viridans]

The effectiveness of antibiotic prophylaxis against endocarditis was assessed by testing the bactericidal and bacteriostatic action in serum of 12 healthy volunteers who had taken the recommended antibiotics according to laid down guidelines. Blood was obtained from these subjects every two hours for 12 hours after oral intake of amoxicillin (3 g as a single dose), clindamycin (600 mg a single dose) or erythromycin (1.5 g plus another 0.5 g after six hours), the serum being tested against three Strep. viridans strains. Two of the three strains were "tolerant", i.e. in relation to the minimal inhibition concentration (MIC) their growth was inhibited, but--in relation to the minimal bactericidal concentration (MBC)--they were not killed. A bacteriostatic effect by amoxicillin and clindamycin was demonstrated by a micromethod against all three strains during the 12-hour period. But erythromycin did not achieve bacteriostasis in all serum samples. A bactericidal effect was demonstrated only in those samples that contained amoxicillin, and then only against the non-tolerant of the three strains. These results support the view that amoxicillin and clindamycin are effective in the prophylaxis against Strep. viridans bacteraemia because of their constant bacteriostatic effect, as measured in serum.

[Perianal erythema in a child--group A streptococci?!]

Perianal streptococcal dermatitis is a common disease. The typical clinical picture includes perianal erythema, pruritus, painful defaecation and bloody stools. The diagnosis is made by a swab taken from the affected skin with bacterial culture. Therapy consists of penicillin for 10 days. Screening for affected persons in contact with the patient is indicated because perianal streptococcal dermatitis is known to be highly contagious. Relapse is common and therefore follow-up visits are recommended. In case of relapse, a first or second generation cephalosporin may be considered.