Orientadores: Jaime A. Cury, Hyun Koo / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-03T18:51:49Z (GMT). No. of bitstreams: 1 Hayacibara_MitsueFujimaki_D.pdf: 743722 bytes, checksum: a03e5c389ce657201cdf9b2b48b9185b (MD5) Previous issue date: 2004 / Doutorado

Orientador: Reginaldo Bruno Gonçalves / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-03T16:11:11Z (GMT). No. of bitstreams: 1 Klein_MarliseInez_M.pdf: 917669 bytes, checksum: d317b17fbe041f50859121aa604f6bed (MD5) Previous issue date: 2003 / Resumo: Os objetivos deste estudo foram avaliar, a partir do momento da colonização inicial da cavidade bucal de crianças por S. mutans e S. sobrinus, a diversidade e a estabilidade de genótipos destas espécies, assim como investigar a ocorrência de transmissão vertical de mãe para filho e/ou horizontal entre crianças que compartilham o mesmo ambiente. Os voluntários foram 16 pares mãe-filho, onde as crianças foram acompanhadas por 19,2 meses em média (menor e maior período de acompanhamento, foram, respectivamente 14,8 e 24,7 meses). Amostras foram coletadas bimestralmente de quatro sítios distintos: saliva, dorso da língua, rodete gengival e placa bacteriana (quando da presença de dentes). Amostras de saliva das mães foram coletada em uma única ocasião. As amostras foram cultivadas sob condições adequadas em meio de cultura MSB (mitis salivarius bacitracina). Em cada colheita foram isoladas colônias com morfologia típica, as quais foram identificadas pela técnica da reação em cadeia da polimerase (PCR). O perfil genético dos isolados identificados como S. mutans e S. sobrinus foi analisado pela técnica de reação em cadeia da polimerase usando primers arbitrários (AP-PCR), utilizando-se os primers arbitrários OPA-02 e OPA-13. Os resultados mostraram que no momento da aquisição, as crianças avaliadas estavam colonizadas por 1 a 4 genótipos distintos de S. mutans (n=16 crianças) e apenas 1 genótipo de S. sobrinus (n=12 crianças). Ocorreu transmissão vertical de S. mutans e de S. sobrinus em 13 e 5 dos 16 pares avaliados, respectivamente. Entretanto, não se detectou ocorrência de transmissão horizontal entre as crianças que compartilhavam o mesmo ambiente. Na avaliação longitudinal observou-se um aumento do número de genótipos que colonizavam a cavidade bucal das crianças, sendo que alguns genótipos de S. mutans e S. sobrinus permaneceram estáveis e outros, idênticos ou não aos da mãe, foram adquiridos. Concluí-se que ocorreu estabilidade e também rotatividade de genótipos de S. mutans e S. sobrinus na cavidade bucal de crianças durante o período avaliado; e que a principal fonte das espécies avaliadas foi a maternal, porém, rotas de transmissão alternativas parecem existir / Abstract: The aims of this study were to evaluate the genotypic diversity and stability of S. mutans and S. sobrinus strains isolated from children oral cavity at time of initial acquisition; and analyze the pattern of vertical transmission of these bacteria from mother to their child among children attending public nursery schools. The subjects were motherchild pairs, and the children were monitored through 19.2 mean months (the smaller and the higher follow-up examination time were 14.8 and 24.7 months, respectively). Samples of four different sites (saliva, tongue dorsum, gengiva, and dental plaque, if teeth were present) were collected bimonthly. Saliva samples from the mothers were collected once. The samples were isolated and cultivated in MSB medium (agar mitis salivarius with bacitracina). Typical morphotype colonies were isolated and submitted to amplification by the technique of polymerase chain reaction (PCR) for identification. The arbitrarily primed polymerase chain reaction (AP-PCR) method was performed with two arbitrary primers, OPA-02 and OPA-13, for the genotypic characterization of S. mutans and S. sobrinus. At the time of acquisition, the children harbored between 1 to 4 distinct genotypes of S. mutans (n=16 children) and only 1 genotype of S. sobrinus (n=12 children). The presence of matching genotypes of S. mutans and S. sobrinus in 13 and 5 from 16 mother-child pairs, respectively, suggested vertical transmission. No horizontal transmission has observed since no matching of S. mutans and/or S. sobrinus genotypes was observed between children attending the same nursery. This longitudinal study showed an increase in genotypic diversity in the oral cavity of children S. mutans and S. sobrinus genotypes, that were or not similar to the mothers have persisted during follow-up and newly strains were also acquired. In conclusion, S. mutans and S. sobrinus stability and turnover occur in children oral cavity during this study; and the major source of this microorganism was maternal, however alternative transmission sources may exist / Mestrado / Microbiologia e Imunologia / Mestre em Biologia Buco-Dental

Made available in DSpace on 2014-06-12T23:01:30Z (GMT). No. of bitstreams: 2 arquivo8544_1.pdf: 706882 bytes, checksum: 7f552d13edf81608cb94ae198a8dccde (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2005 / Neste estudo foi investigada a capacidade antimicrobiana in vitro do extrato hidroalcoólico de alecrim (Rosmarinus officinalis Linn.) sobre bactérias orais planctônicas. Foram utilizados o extrato puro, suas diluições de 1/2 à 1/512, clorexidina e cepas padrão de Streptococcus mitis ATCC 98811, Streptococcus sanguinis ATCC 10556, Streptococcus mutans ATCC 25175, Streptococcus sobrinus ATCC 27609 e Lactobacillus casei ATCC 7469. Inicialmente, foi determinada a CIM do extrato de alecrim pelo método de difusão em meio sólido sobre as cepas supracitadas. Em seguida, foi determinada a CIMA do extrato sobre as cepas padrão de Streptococcus mitis ATCC 98811, Streptococcus sanguinis ATCC 10556, Streptococcus mutans ATCC 25175, Streptococcus sobrinus ATCC 27609, na presença de sacarose a 5%. Todos os ensaios foram realizados em duplicata e a clorexidina foi utilizada como solução controle. A CIM variou de 66,5 mg/mL a 266 mg/mL e a CIMA de 16 mg/mL a 33,2 mg/mL. Foi observado que o extrato hidroalcoólico puro de alecrim e diluído até 1/16 apresentou ação antimicrobiana sobre as cepas padrão ensaiadas, exceto sobre Streptococcus mitis ATCC 98811. Não foi observado efeito inibitório de aderência sobre Streptococcus sanguinis ATCC 10556. Streptococcus mutans ATCC 25175 e Lactobacillus casei ATCC 7469 foram as cepas mais sensíveis à ação antimicrobiana in vitro do extrato de alecrim

Tesis para optar al grado de Magíster en Bioquímica área de Especialización en Bioquímica Clínica / Streptococcus agalactiae (SGB) es una bacteria Gram positiva anaeróbica facultativa capaz de colonizar el tracto genitourinario de un 15 a un 40% de mujeres sexualmente activas. En la mayoría de los casos la infección con este patógeno es asintomática pero en neonatos puede originar sepsis, meningitis y neumonía. A partir del año 1990 el CDC (Center for Disease Control and Prevention) ha implementado una serie de protocolos de prevención dirigidos a mujeres embarazadas los cuales constan de pruebas microbiológicas realizadas entre las 35 y 37 semanas de gestación y la aplicación de antibióticos intraparto en mujeres colonizadas. A pesar de que medidas han tenido un alto impacto en la disminución de la incidencia de la infección por SGB este patógeno persiste como el único y más frecuente aislado desde la sangre o del fluido cerebroespinal de infantes menores de 3 meses de edad y de mujeres con infecciones intraparto en Estados Unidos y en otros países industrializados. Numerosos estudios realizados a neonatos sanos nacidos de madres colonizadas con SGB indican que estos tienen niveles superiores de anticuerpos IgG transferidos transplacentalmente que aquellos que desarrollaron la infección. Basado en este hecho actualmente se trabaja arduamente en el desarrollo de una vacuna capaz proteger a mujeres embarazadas y sus infantes. En la actualidad, vacunas en base a polisacáridos conjugados están en Fase clínica III, con el inconveniente de que la vacuna ensayada será serotipo dependiente y solamente inducirá una respuesta inmune protectora contra los serotipos incluidos en su formulación (3 de 10 serotipos). Vacunas en base a proteínas conservadas de SGB se encuentran en estudio, una de ellas es la proteína SIP (Surface Immunogenic Protein), con la cual se trabaja en la Sección de Biotecnología del ISPCH. En este estudio se evaluará la respuesta inmune protectora de un prototipo de vacuna en base a la proteína recombinante SIP de SGB, evaluando parámetros inmunológicos que permitan demostrar la inducción de inmunidad protectora en modelo ratón

BACKGROUND AND AIMS OF THE THESIS: Group A β-haemolytic Streptococcus (GAS) also known as Streptococcus pyogenes, is responsible for a wide range of invasive and non-invasive GAS diseases. Prevalence and incidence data on GAS from developing countries are largely lacking when compared with industrialised nations. This thesis sought to (1) establish the South African arm of the AFROStrep biorepository and clinical database for patients with invasive and non-invasive GAS infection, (2) identify and summarize all published studies of laboratory-confirmed GAS infection in Africa, (3) describe, from national laboratory data, the incidence of invasive and non-invasive GAS in South Africa and, (4) conduct a prospective, surveillance study in order to determine the molecular epidemiology of GAS in Cape Town, South Africa over a 12-month period. METHODS: A systematic review was conducted on population-based studies reporting on the prevalence of laboratory-confirmed GAS infection among patients living in Africa (Study 1). A retrospective study of the incidence of GAS infection was conducted on data obtained from the National Health Laboratory Service between 2003 – 2015 (Study 2). The AFROStrep registry and biorepository (based in Cape Town) was established and through passive surveillance, laboratory confirmed invasive and non-invasive GAS cases were collected over a 12-month period. The molecular analysis of invasive and non-invasive infection was determined using emm type sequencing to provide insight into vaccine development (Study 3). RESULTS AND DISCUSSION: The pooled prevalence of GAS pharyngitis in Africa was determined to be 21% (95% CI, 17% to 26%). The incidence rates of laboratory-confirmed non-invasive GAS infection in the South African public sector appears to have declined over the last 13 years. Given the possibility that the lower incidence of invasive and non-invasive GAS infection found in our study is due to infrequent submission of specimens for microbiological culture by health practitioners, our findings may be an underestimate of the true burden of disease in South Africa. In our prospective surveillance study, 46 different emm types were identified. The most prevalent emm types were M76 (16% of isolates), M81 (10%), M80 (6%), M43 (6%), and M183 (6%) and were almost evenly distributed between invasive and non-invasive GAS isolates. When compared against the putative 30-valent vaccine under development, four of our most prevalent emm types are not included; vaccine coverage (i.e. vaccine type and non-vaccine type-killing) for non-invasive and invasive GAS infection in our setting was 60% and 59% respectively, notably lower than coverage in developed countries. CONCLUSION: This work provides evidence for a significantly high prevalence of GAS pharyngitis in Africa. While GAS surveillance in South Africa indicates a declining incidence of GAS disease in parts of the country over the last thirteen years, the findings may be an underestimate of the true burden of disease, demonstrating the need for accurate and comprehensive surveillance of GAS in South Africa. Finally, this research showed a low potential vaccine coverage in our setting and thus, emphasises the need for a reworking of the potential vaccine formulation to improve coverage in areas where the burden of disease is high.

Streptococcus pneumoniae is a bacterium responsible for a spectrum of diseases including lobar pneumonia, meningitis, otitis media, and sinusitis. Invasive pneumococcal disease is responsible for significant morbidity and mortality across the world. Concerted efforts led to the development of two vaccinations, Pneumova × 23 and Prevnar 13, for the prevention of pneumococcal disease. The Advisory Committee on Immunization Practices of the US Centers for Disease Control and Prevention provides vaccination schedules for predisposed adults, but the proposed schedules remain a challenge to health-care providers. We performed a systematic review in PubMed and these specialty group websites to present the pathophysiology of pneumococcal disease, outline different pneumococcal vaccinations, and condense recommendations for vaccination administration.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

No description available.

Streptococcus pneumoniae is an asymptomatic colonizer of the upper respiratory tract as well as an opportunistic pathogen. Colonization is prerequisite to causing disease in a host, and it often involves formation of biofilms. There are currently two vaccines available against pneumococcus, both of which focus on preventing invasive disease by targeting the polysaccharide capsule of the most invasive serotypes. We hypothesized that by using membrane proteins expressed during the biofilm state, we can formulate an effective vaccine against colonization which would lead to an overall decrease in disease incidence. To do this, we selected protein candidates expressed during biofilm growth based on their ability to elicit an IgA response in human serum. Selected proteins (SP_0459, SP_1114, and SP_1702) were identified and used for further experiments. The proteins identified in this study will be paired with other immunogenic proteins to determine a successful vaccine formulation targeting colonization of Streptococcus pneumoniae.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.