Induction of neutralizing antibodies in mice immunized with an amino-terminal polypeptide of Streptococcus mutans P1 protein produced by a recombinant Bacillus subtilis strain

The oral pathogen Streptococcus mutans expresses a surface protein, P1, which interacts with the salivary pellicle on the tooth surface or with fluid-phase saliva, resulting in bacterial adhesion or aggregation, respectively. P1 is a target of protective immunity. Its N-terminal region has been associated with adhesion and aggregation functions and contains epitopes recognized by efficacious antibodies. In this study, we used Bacillus subtilis, a gram-positive expression host, to produce a recombinant N-terminal polypeptide of P1 (P1(39-512)) derived from the S. mutans strain UA159. Purified P1(39-512) reacted with an anti-full-length P1 antiserum as well as one raised against intact S. mutans cells, indicating preserved antigenicity. Immunization of mice with soluble and heat-denatured P1(39-512) induced antibodies that reacted specifically with native P1 on the surface of S. mutans cells. The anti-P1(39-512) antiserum was as effective at blocking saliva-mediated aggregation of S. mutans cells and better at blocking bacterial adhesion to saliva-coated plastic surfaces compared with the anti-full-length P1 antiserum. In addition, adsorption of the anti-P1 antiserum with P1(39-512) eliminated its ability to block the adhesion of S. mutans cells to abiotic surfaces. The present results indicate that P1(39-512), expressed and purified from a recombinant B. subtilis strain, maintains important immunological features of the native protein and represents an additional tool for the development of anticaries vaccines.

Longitudinal study of the influence of removable partial denture and chemical control on the levels of Streptococcus mutans in saliva

Some studies have evaluated the salivary levels of mutans streptococci (MS) in removable partial denture (RPD) users. Saliva samples (2.0 mL) were obtained from 31 patients in six periods: (T0): immediately before installation of RPD; (T8): 8 days after T0; (T48): 48 days after T0; (T92): 92 days after T0; (T140): 140 days after T0 and (T189): 189 days after T0. The samples were vortexed and serially diluted from 10(-1) to 10(-6) in 0.05 m phosphate buffer (pH 7.4). From each dilution, 0.025 mL was plated on Mitis Salivarius Bacitracin (MSB). The plates were incubated in 5% CO2 at 37 degreesC for 72 h. There was an increase (t -test, P < 0.05) in the number of MS between periods T0 and T48 (mean/s.d., CFU mL(-1) of saliva): T0: 2.26/4.43 x 10(6) and T48: 0.47/1.48 x 10(8) . After this, intensive treatment with CHX was accomplished in 29 patients. Saliva samples were obtained after treatment in four periods: (T24 h): 24 h after T0; (T14): 14 days after T24 h; (T28): 28 days after T24 h, and (T63): 63 days after T24 h. The number of MS in saliva did not decrease (t -test, P > 0.05). A new CHX formulation was applied in 15 patients. Saliva samples were obtained in periods: (T0): before new CHX application; (T24 h): 24 h after T0 and (T82): 82 days after T0. The new CHX reduced MS levels in saliva: (mean/s.d., CFU mL(-1) of saliva): T0: 6.64/8.47 x 10(6) and T24 h: 3.2/4.27 x 10(5) (sign rank, P < 0.05). In conclusion, there was a significant increase in the number of MS in saliva after the installation of RPD. The intensive treatment with a properly formulated CHX was effective in the reduction of MS, between 24 h and 82 days after its application.

Effect of Psidium cattleianum leaf extract on Streptococcus mutans viability, protein expression and acid production

Plants naturally produce secondary metabolites that can be used as antimicrobials. The aim of this study was to assess the effects of Psidium cattleianum leaf extract on Streptococcus mutans. The extract (100%) was obtained by decoction of 100 g of leaves in 600 ml of deionized water. To assess killing, S. mutans biofilms were treated with water (negative control) or various extract dilutions [ 100, 50, 25% (v/v) in water] for 5 or 60 min. To evaluate the effect on protein expression, biofilms were exposed to water or 1.6% (v/v) extract for 120 min, proteins were extracted and submitted to 2-dimensional difference gel electrophoresis. Differentially expressed proteins were identified by mass spectrometry. The effect of 1.6% (v/v) extract on acid production was determined by pH measurements and compared to a water control. Viability was similar after 5 min of treatment with the 100% extract or 60 min with the 50% extract (about 0.03% survival). There were no differences in viability between the biofilms exposed to the 25 or 50% extract after 60 min of treatment (about 0.02% survival). Treatment with the 1.6% extract significantly changed protein expression. The abundance of 24 spots was decreased compared to water (p < 0.05). The extract significantly inhibited acid production (p < 0.05). It is concluded that P. cattleianum leaf extract kills S. mutans grown in biofilms when applied at high concentrations. At low concentrations it inhibits S. mutans acid production and reduces the expression of proteins involved in general metabolism, glycolysis and lactic acid production. Copyright (C) 2008 S. Karger AG, Basel.

Influence of sample type and collection method on Streptococcus mutans and Lactobacillus spp. counts in the oral cavity

Objectives: This study compared three methods of Streptococcus mutans and Lactobacillus spp. detection in the oral cavity: saliva swab (SS)-sample of stimulated saliva collected with swab; whole saliva (WS)-sample of 2 ml of stimulated saliva; and the dental plaque method (DP)-plaque sample of all dental surfaces.Methods: Thirty children were included in this study. In the first 15 children, the SS and WS methods were carried out before the dental plaque collection, and in the following 15, the sequence was inverted to evaluate possible interference of the methods sequence. The samples were diluted and inoculated in SB20 and Rogosa agar, respectively for S. mutans and Lactobacillus spp., at 37 degrees C for 48 h.Results: the results (cfu/mL) of S. mutans were analysed by the statistical Friedman's test. The levels of Lactobacillus spp. were analysed by descriptive statistics due to the high proportion of zero counts in the culture. In the first sequence of methods, the number of S. mutans counted for the SS method was inferior to DP and WS (P < 0.05), and the results for the WS and DP methods were similar. The detection of Lactobacillus spp. was observed just by the WS (100 %) and SS (14.3 %) methods. However, in the second experimental set the number of S. mutans detected by the DP method was similar to those of the SS and WS, however, the WS method showed higher values than SS (P < 0.05). A greater number of Lactobacillus spp. was detected by the WS method (100 %), followed by SS (55.5 %) and DP (33.3 %).Conclusions: the dental plaque collection and the sample of stimulated whole saliva presented similar results in the S. mutans count. The most suitable method to detect the Lactobacillus spp. level in the oral cavity is the stimulated whole saliva method. (c) 2004 Elsevier Ltd. All rights reserved.

Fatigue and Fluoride Corrosion on Streptococcus mutans Adherence to Titanium-Based Implant/Component Surfaces

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Genetic polymorphism of Streptococcus mutans in Brazilian family members

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Caries risk tests and salivary levels of immunoglobulins to Streptococcus mutans and Candida albicans in mouthbreathing syndrome patients

The aim of this study was to compare microbiological and salivary variables possibly related to caries risk in treated and untreated mouthbreathing syndrome (MBS) children and control children. Thirty control children, 30 mouthbreathers and 25 treated mouthbreathers were studied for the numbers of lactobacilli, mutans streptococci and yeasts in their saliva. Snyder's test, salivary flow and buffering capacity were also evaluated. Levels of immunoglobulins to Candida albicans and Streptococcus mutans in the saliva were quantified using ELISA. Considering the results obtained for the microbiological and salivary caries risk tests, no significant differences were observed among the proportions of patients with small/negative and high/moderate caries risk in the studied groups. The level of IgG to S. mutans was significantly higher in the treated MBS group in relation to MBS patients. on the other hand, the median anti-S. mutans IgM level was lower in the treated MBS patients than in the other groups. For the studied anti-Candida immunoglobulins, IgM level was significantly lower in the treated MBS group than in the other groups. No differences were observed for anti-S. mutans and anti-Candida IgA levels among the groups. The findings suggest that mouthbreathing cannot be considered a risk factor for dental caries. Copyright (C) 2003 S. KargerAG, Basel.

Susceptibility of Candida albicans, Staphylococcus aureus, and Streptococcus mutans biofilms to photodynamic inactivation: an in vitro study

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Streptococcus mutans Biofilm Adhesion on Composite Resin Surfaces After Different Finishing and Polishing Techniques

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

In Vitro Antibacterial Activity of Adhesive Systems on Streptococcus Mutans

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Detecção de anticorpo anticoração em camundongos Balb/c imunizados com Streptococcus mutans

Anticorpos para antígenos cardíacos foram analisados por ELISA em 14 soros de camundongos Balb/c hiperimunizados com Streptococcus mutans, inativado pelo formaldeído. Os níveis de anticorpos da classe IgG anticoração e antimiosina elevaram-se significativamente nos animais imunizados quando comparados com os controles, especialmente no grupo A, imunizado e reestimulado com antígenos solúveis de S. mutans. Neste grupo, os resultados do Western Blot mostraram reatividade com miosina cardíaca e uma banda de 35 kDa. A análise histológica dos corações dos animais do grupo B, imunizado e reestimulado com antígenos de superfície do microrganismo, demonstrou a presença de degeneração celular, tipo hidrópica e hialina e focos inflamatórios constituídos de linfócitos e macrófagos no miocárdio e pericárdio. Os resultados deste trabalho reforçam a hipótese da existência de mimetismo antigênico entre tecido cardíaco e S. mutans e chamam a atenção para o risco de desenvolvimento de anticorpos reativos com antígenos próprios induzidos por vacina anticárie com componentes estreptocócicos.

IgA anti-Streptococcus mutans em crianças com e sem cárie dentária

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of oral environment stabilization procedures on Streptococcus mutans counts in pregnant women

O objetivo desta pesquisa foi determinar o efeito da adequação do meio bucal sobre a contagem de Streptococcus mutans em gestantes de alto risco à cárie, participantes de um programa de prevenção em uma instituição de ensino público, antes e após adequação do meio bucal. Amostras de saliva de 30 gestantes (18 a 43 anos) que procuraram atendimento na Clínica de Odontologia Preventiva da FOAr-UNESP foram coletadas antes e após procedimentos de adequação do meio bucal, e examinadas para observação e contagem das UFCs de S. mutans. Foi demonstrado que houve diminuição na quantidade das UFCs (p<0,0001) entre as amostras. em relação à faixa etária, 70,0% das gestantes tinham entre 18 e 30 anos de idade e 30,0% pertenciam à faixa etária de 31 a 43 anos. Dados relativos ao período da gestação revelaram que 73,4% estavam no 2º trimestre e 13,3% estavam igualitariamente no 1º e 3º trimestres. A adequação do meio bucal se mostrou eficaz na diminuição das UFCs de S. mutans presentes na saliva de gestantes de alto risco de cárie. Este procedimento é simples e extremamente efetivo, atendendo às necessidades de tratamento básico de gestantes que procuram atendimento odontológico no serviço público de saúde.