Contaminação cruzada em escovas dentais por Streptococcus mutans

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Aderência de Streptococcus mutans em materiais restauradores e seus efeitos em cárie secundária in vitro E in situ avaliados por microscopia de luz polarizada e espectroscopia por dispersão de raios-X (EDS)

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

Pesquisa de Streptococcus Agalactiae em gestantes residentes em Belém-Pará

Na infecção por Streptococcus agalactiae são reconhecidas duas formas neonatais, a de início precoce, cujo quadro clínico é caracterizado por bacteremia com envolvimento pulmonar, meningite é a manifestação clínica predominante. Considerando-se a gravidade da patologia, o desconhecimento da incidência desta bactéria em gestantes residentes na região Norte do Brasil e a importância do seu diagnóstico em exames pré-natais, é fundamental a determinação da ocorrência dos estreptococos do B neste referido grupo populacional. Portanto, este estudo objetivou a realização do diagnóstico laboratorial de Streptococcus agalactiae no trato genital feminino de gestantes, no último trimestre de gestação, determinando a incidência e alertando sobre a importância do diagnóstico no exame pré-natal. O estudo foi realizado no período de fevereiro a agosto de 2002, em 50 gestantes voluntárias residentes e domiciliadas na cidade de Belém-Pará, procedentes do setor de Tocoginecologia da Universidade Federal do Pará, e a identificação da bactéria foi realizada através da bacterioscopia e da cultura do conteúdo vaginal. Das 50 gestantes estudadas, sete (14 %) apresentavam cultura positiva para Streptococcus agalactiae. Destas, duas (28,6 %) eram primigestas e cinco (71,4 %) secundigestas. Os resultados obtidos indicaram a presença significativa da bactéria, indicando a necessidade da adoção de medidas profiláticas da infecção por este agente, devido às altas taxas de morbidade e mortalidade associadas ao estreptococo do grupo B.

Topografia superficial e aderência de Streptococcus mutans no titânio após imersão e escovação com dentifrícios fluoretado e não-fluoretado simulando 10 anos de uso

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo da eficácia da terapia fotodinâmica, mediada pelos fotossensibilizadores Photodithazine® e Curcumina, sobre biofilmes multi-espécies formados por Streptococcus mutans, Candida albicans e Candida glabrata

Pós-graduação em Reabilitação Oral - FOAR

Resposta imune de tilápias do Nilo (Oreochromis niloticus) vacinadas contra Streptococcus agalactiae

Pós-graduação em Medicina Veterinária - FCAV

Ocorrência, sorotipagem e rsistência antimicrobiana de Streptococcus suis em funcionários e ambente de abatedouro de sínos

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

Efeito de soluções fluoretadas contendo xilitol e sorbitol no número de estreptococos do grupo mutans na saliva de seres humanos

The objective of this study was to assess the effect of 0.05% sodium fluoride solutions containing 2.5% or 12.5% xylitol on the number of Streptococcus mutans in the human mouth. Fifty boys between 8 and 16 years of age participated in this double-blind crossover study. Of the original 50 boys, 33 finished the study. Participants were randomly divided into four groups. The following solutions were employed: placebo solution; 0.05% sodium fluoride solution; 0.05% sodium fluoride + 2.5% xylitol + 2% sorbitol; 0.05% sodium fluoride + 12.5% xylitol + 2% sorbitol. Each solution was used for a 28-day period (20 mL/day, twice a day), with a 10-day washout period between solutions. There were no significant differences (P = 0.32) between the two xylitol-containing solutions (2.5% vs. 12.5%) concerning the number of Streptococcus mutans. However, there was a significant difference between these two xylitol-containing solutions and the sodium fluoride and placebo solutions (P < 0.001). Our results suggest that the 0.05% sodium fluoride solutions containing either 2.5% or 12.5% xylitol caused a significant reduction in the number of Streptococcus mutans.

Imunohistoquímica em órgãos de tilápias-do-Nilo imunizadas com antígeno insolúvel de Streptococcus agalactiae

Pós-graduação em Medicina Veterinária - FCAV

Inativação fotodinâmica em biofilme de Streptococcus mutans sobre bráquetes metálicos e cerâmicos: um estudo in vitro

Pós-graduação em Odontologia Restauradora - ICT

Revisiting molecular diagnostics of Streptococcus pneumoniae

Streptococcus pneumoniae is a human pathobiont that colonizes the nasopharynx. S. pneumoniae is responsible for causing non-invasive and invasive disease such as otitis, pneumonia, meningitis, and sepsis, being a leading cause of infectious diseases worldwide. Due to similarities with closely related species sharing the same niche, it may be a challenge to correctly distinguish S. pneumoniae from its relatives when using only non-culture based methods such as real time PCR (qPCR). In 2007, a molecular method targeting the major autolysin (lytA) of S. pneumoniae by a qPCR assay was proposed by Carvalho and collaborators to identify pneumococcus. Since then, this method has been widely used worldwide. In 2013, the gene encoding for the ABC iron transporter lipoprotein PiaA, was proposed by Trzcinzki and collaborators to be used in parallel with the lytA qPCR assay. However, the presence of lytA gene homologues has been described in closely related species such as S. pseudopneumoniae and S. mitis and the presence of piaA gene is not ubiquitous between S. pneumoniae. The hyaluronate lyase gene (hylA) has been described to be ubiquitous in S. pneumoniae. This gene has not been used so far as a target for the identification of S. pneumoniae. The aims of our study were to evaluate the specificity, sensitivity, positive predicted value (PPV) and negative predicted value (NPV) of the lytA and piaA qPCR methods; design and implement a new assay targeting the hylA gene and evaluate the same parameters above described; analyze the assays independently and the possible combinations to access what is the best approach using qPCR to identify S. pneumoniae. A total of 278 previously characterized strains were tested: 61 S. pseudopneumoniae, 37 Viridans group strains, 30 type strains from other streptococcal species and 150 S. pneumoniae strains. The collection included both carriage and disease isolates. By Mulilocus Sequence Analysis (MLSA) we confirmed that strains of S. pseudopneumoniae could be misidentified as S. pneumoniae when lytA qPCR assay is used. The results showed that as a single target, lytA had the best combination of specificity, sensitivity, PPV and NPV being, 98.5%, 100.0%, 98.7% and 100.0% respectively. The combination of targets with the best values of specificity, sensibility, PPV and NPV were lytA and piaA, with 100.0%, 93.3%, 97.9% and 92.6%, respectively. Nonetheless by MLSA we confirmed that strains of S. pseudopneumoniae could be misidentified as S. pneumoniae and some capsulated (23F, 6B and 11A) and non-capsulated S. pneumoniae were not Identified using this assay. The hylA gene as a single target had the lowest PPV. Nonetheless it was capable to correctly identify all S. pneumoniae.

Immunogenicity and protective efficacy of an anti-Streptococcus pyogenes vaccine candidate in multiple animal species

Streptococcus pyogenes, also known as Group A Streptococcus (GAS) has been associated with a range of diseases from the mild pharyngitis and pyoderma to more severe invasive infections such as streptococcal toxic shock. GAS also causes a number of non-suppurative post-infectious diseases such as rheumatic fever, rheumatic heart disease and glomerulonephritis. The large extent of GAS disease burden necessitates the need for a prophylactic vaccine that could target the diverse GAS emm types circulating globally. Anti-GAS vaccine strategies have focused primarily on the GAS M-protein, an extracellular virulence factor anchored to GAS cell wall. As opposed to the hypervariable N-terminal region, the C-terminal portion of the protein is highly conserved among different GAS emm types and is the focus of a leading GAS vaccine candidate, J8-DT/alum. The vaccine candidate J8-DT/alum was shown to be immunogenic in mice, rabbits and the non-human primates, hamadryas baboons. Similar responses to J8-DT/alum were observed after subcutaneous and intramuscular immunization with J8-DT/alum, in mice and in rabbits. Further assessment of parameters that may influence the immunogenicity of J8-DT demonstrated that the immune responses were identical in male and female mice and the use of alum as an adjuvant in the vaccine formulation significantly increased its immunogenicity, resulting in a long-lived serum IgG response. Contrary to the previous findings, the data in this thesis indicates that a primary immunization with J8-DT/alum (50ƒÊg) followed by a single boost is sufficient to generate a robust immune response in mice. As expected, the IgG response to J8- DT/alum was a Th2 type response consisting predominantly of the isotype IgG1 accompanied by lower levels of IgG2a. Intramuscular vaccination of rabbits with J8-DT/alum demonstrated that an increase in the dose of J8-DT/alum up to 500ƒÊg does not have an impact on the serum IgG titers achieved. Similar to the immune response in mice, immunization with J8-DT/alum in baboons also established that a 60ƒÊg dose compared to either 30ƒÊg or 120ƒÊg was sufficient to generate a robust immune response. Interestingly, mucosal infection of naive baboons with a M1 GAS strain did not induce a J8-specific serum IgG response. As J8-DT/alum mediated protection has been previously reported to be due to the J8- specific antibody formed, the efficacy of J8-DT antibodies was determined in vitro and in vivo. In vitro opsonization and in vivo passive transfer confirmed the protective potential of J8-DT antibodies. A reduction in the bacterial burden after challenge with a bioluminescent M49 GAS strain in mice that were passively administered J8-DT IgG established that protection due to J8-DT was mediated by antibodies. The GAS burden in infected mice was monitored using bioluminescent imaging in addition to traditional CFU assays. Bioluminescent GAS strains including the ‘rheumatogenic’ M1 GAS could not be generated due to limitations with transformation of GAS, however, a M49 GAS strain was utilized during BLI. The M49 serotype is traditionally a ‘nephritogenic’ serotype associated with post-streptococcal glomerulonephritis. Anti- J8-DT antibodies now have been shown to be protective against multiple GAS strains such as M49 and M1. This study evaluated the immunogenicity of J8-DT/alum in different species of experimental animals in preparation for phase I human clinical trials and provided the ground work for the development of a rapid non-invasive assay for evaluation of vaccine candidates.

Streptococcus pneumoniae infection suppresses allergic airways disease by inducing regulatory T cells

An inverse association exists between some bacterial infections and the prevalence of asthma. We investigated whether Streptococcus pneumoniae infection protects against asthma using mouse models of ovalbumin (OVA)-induced allergic airway disease (AAD). Mice were intratracheally infected or treated with killed S. pneumoniae before, during or after OVA sensitisation and subsequent challenge. The effects of S. pneumoniae on AAD were assessed. Infection or treatment with killed S. pneumoniae suppressed hallmark features of AAD, including antigen-specific T-helper cell (Th) type 2 cytokine and antibody responses, peripheral and pulmonary eosinophil accumulation, goblet cell hyperplasia, and airway hyperresponsiveness. The effect of infection on the development of specific features of AAD depended on the timing of infection relative to allergic sensitisation and challenge. Infection induced significant increases in regulatory T-cell (Treg) numbers in lymph nodes, which correlated with the degree of suppression of AAD. Tregs reduced T-cell proliferation and Th2 cytokine release. The suppressive effects of infection were reversed by anti-CD25 treatment. Respiratory infection or treatment with S. pneumoniae attenuates allergic immune responses and suppresses AAD. These effects may be mediated by S. pneumoniae-induced Tregs. This identifies the potential for the development of therapeutic agents for asthma from S. pneumoniae.

Evaluation of novel Streptococcus pyogenes vaccine candidates incorporating multiple conserved sequences from the C-repeat region of the M-protein

A major challenge for Streptococcus pyogenes vaccine development is the identification of epitopes that confer protection from infection by multiple S. pyogenes M-types. Here we have identified and characterised the distribution of common variant sequences from individual repeat units of the C-repeat region (CRR) of M-proteins representing 77 different M-types. Three polyvalent fusion vaccine candidates (SV1, SV2 and SV3) incorporating the most common variants were subsequently expressed and purified, and demonstrated to be alpha-helical by Circular Dichroism (CD), a secondary conformational characteristic of the CRR in the M-protein. Antibodies raised against each of these constructs recognise M-proteins that vary in their CRR, and bind to the surface of multiple S. pyogenes isolates. Antibodies raised against SV1, containing five variant sequences, also kill heterologous S. pyogenes isolates in in vitro bactericidal assays. Further structural characterisation of this construct demonstrated the conformation of SV1 was stable at different pHs, and thermal unfolding of SV1 a reversible process. Our findings demonstrate that linkage of multiple variant sequences into a single recombinant construct overcomes the need to embed the variant sequences in foreign helix promoting flanking sequences for conformational stability, and demonstrates the viability of the polyvalent candidates as global S. pyogenes vaccine candidates.