The Biology of Streptococcus mutans.

As a major etiological agent of human dental caries, Streptococcus mutans resides primarily in biofilms that form on the tooth surfaces, also known as dental plaque. In addition to caries, S. mutans is responsible for cases of infective endocarditis with a subset of strains being indirectly implicated with the onset of additional extraoral pathologies. During the past 4 decades, functional studies of S. mutans have focused on understanding the molecular mechanisms the organism employs to form robust biofilms on tooth surfaces, to rapidly metabolize a wide variety of carbohydrates obtained from the host diet, and to survive numerous (and frequent) environmental challenges encountered in oral biofilms. In these areas of research, S. mutans has served as a model organism for ground-breaking new discoveries that have, at times, challenged long-standing dogmas based on bacterial paradigms such as Escherichia coli and Bacillus subtilis. In addition to sections dedicated to carbohydrate metabolism, biofilm formation, and stress responses, this article discusses newer developments in S. mutans biology research, namely, how S. mutans interspecies and cross-kingdom interactions dictate the development and pathogenic potential of oral biofilms and how next-generation sequencing technologies have led to a much better understanding of the physiology and diversity of S. mutans as a species.

Whole genome sequence and phenotypic characterization of a Cbm+ serotype e strain of Streptococcus mutans.

We report the whole genome sequence of the serotype e Cbm+ strain LAR01 of Streptococcus mutans, a dental pathogen frequently associated with extra-oral infections. The LAR01 genome is a single circular chromosome of 2.1 Mb with a GC content of 36.96%. The genome contains 15 phosphotransferase system gene clusters, seven cell wall-anchored (LPxTG) proteins, all genes required for the development of natural competence and genes coding for mutacins VI and K8. Interestingly, the cbm gene is genetically linked to a putative type VII secretion system that has been found in Mycobacteria and few other Gram-positive bacteria. When compared with the UA159 type strain, phenotypic characterization of LAR01 revealed increased biofilm formation in the presence of either glucose or sucrose but similar abilities to withstand acid and oxidative stresses. LAR01 was unable to inhibit the growth of Strpetococcus gordonii, which is consistent with the genomic data that indicate absence of mutacins that can kill mitis streptococci. On the other hand, LAR01 effectively inhibited growth of other S. mutans strains, suggesting that it may be specialized to outcompete strains from its own species. In vitro and in vivo studies using mutational and heterologous expression approaches revealed that Cbm is a virulence factor of S. mutans by mediating binding to extracellular matrix proteins and intracellular invasion. Collectively, the whole genome sequence analysis and phenotypic characterization of LAR01 provides new insights on the virulence properties of S. mutans and grants further opportunities to understand the genomic fluidity of this important human pathogen.

Chemical composition and antifungal potential of Brazilian propolis against Candida spp.

Propolis is known to have biological properties against numerous microorganisms of clinical interest. This study aimed to determine the chemical composition and antifungal activity of Brazilian propolis (types 3 and 13) against Candida spp. and their effects on the morphology of preformed and mature Candida biofilms. Samples of propolis (3 and 13) collected by Apis mellifera honeybees were obtained from different regions in Brazil. Ethanolic extracts of propolis (EEP) were prepared, fractionated and submitted to chemical analysis by GC/MS. The extracts and their hexane, dichloromethane and ethyl acetate fractions were tested for their ability to inhibit Candida spp. (C. albicans, C. dubliniensis, C. glabrata, C. kruzei, C. tropicalis and C. parapsilosis) by determination of the minimum inhibitory and fungicidal concentrations (MIC/MFC). Additionally, their effects on morphology of preformed and mature biofilms were observed by scanning electron microscopy. The phenolic compounds p-coumaric acid, caffeic acid phenethyl ester (CAPE), kaempferol and quercetin were identified in the EEP-3 and its bioactive dichloromethane fraction; and isoflavonoids such as medicarpin, vestitol and formononetin were found in the EEP-13, and triterpenes in its bioactive hexane fraction. The EEP-3 and EEP-13 and their bioactive fractions showed MIC values ranging from 0.2 to 125µg/mL and MFC values between 125 and 500µg/mL. The EEP and fractions were predominantly fungistatic agents. All extracts and fractions disrupted biofilm structures at 500µg/mL and amorphous areas with cell damage were clearly observed in preformed and mature biofilms. Propolis types 3 and 13 have strong anti-Candida activity and should be considered as promising candidates to treat oral and systemic candidiasis.

Clinical applicability of natural product(s)-containing mouthwashes as adjunctive treatment of biofilm-induced gingivitis: a systematic review / Aplicabilidade clínica de colutórios à base de produtos naturais como tratamento adjuvante da gengivite induzida por biofilme: uma revisão sistemática

Natural products have emerged as an effective and low-cost alternative for treating various diseases of the oral cavity. This study aimed to evaluate, through a systematic literature review, if there is scientific evidence ensuring the safe and effective use of natural product(s)-containing mouthwashes as adjunctive treatment of biofilm-induced gingivitis. Searches were conducted in the databases Medline, SciELO, LILACS and Cochrane Library, by using combinations of the key words gingivitis/natural products/phytotherapy/mouthwash, in English, Portuguese and Spanish. Studies published until September 2010 were considered. Four examiners analyzed independently: study design and phase, methodological quality (Jadad scale - JE), experimental product and its concentration, dosing interval and time of usage, as well as employed statistical analysis and clinical outcome of interest. From the 503 articles found, 08 were included in the final review as phase II, controlled, randomized and blind clinical trials, scoring 4 (25%) and 5 (75%) in JE. The main natural products assessed were: Azadirachta indica, Garcinia mangostana, Lippia sidoides, Salvadora persica and Sesamum indicum whose concentration, dosing interval, time of usage and adverse effects varied according to each study. The Plaque and Gingival Index were most employed, as well as α = 5% and paired t, Student's t, Wilcoxon and Mann-Whitney tests. A total of 62.5% and 50% of the products significantly reduced supragingival biofilm and gingivitis, respectively. Mouthwashes containing the essential oil from the leaves of L. sidoides (1%) and the extract from the leaves of A. indica (25%) can be indicated as adjunctive treatment of biofilm-induced gingivitis.

Os produtos naturais têm surgido como alternativa eficaz e de baixo custo para o tratamento de várias doenças da cavidade oral. Objetivou-se avaliar, a partir de revisão sistemática da literatura, se há evidências científicas garantindo a utilização segura e eficaz de antissépticos bucais contendo produto(s) natural(is) como tratamento adjuvante da gengivite induzida por biofilme. Foram realizadas buscas nas bases de dados Medline, SciELO, LILACS e Cochrane Library, através de combinações usando as palavras-chave gengivite/produtos naturais/ fitoterápicos/bochechos, em Inglês, Português e Espanhol. Consideraram-se os estudos publicados até setembro de 2010. Quatro examinadores analisaram separadamente: desenho e fase do estudo, qualidade metodológica (escala de Jadad - EJ), produto experimental e a concentração, intervalo de administração e tempo de uso, bem como a análise estatística empregada e os resultados clínicos de interesse. Foram encontrados 503 artigos dos quais 08 foram incluídos na revisão final como sendo ensaios clínicos fase II, controlados, randomizados e cegos, marcando 4 (25%) e 5 (75%) na EJ. Os principais produtos naturais avaliados foram Azadirachta indica, Garcinia mangostana, Lippia sidoides, Salvadora persica e Sesamum indicum, cuja concentração, intervalo de administração, tempo de uso, e efeitos adversos, variaram de acordo com cada estudo. Índice de placa e Índice Gengival foram os mais utilizados, bem como α=5% e testes t-pareado, t-Student, Wilcoxon e Mann-Whitney. 62,5% e 50% dos produtos reduziram significativamente a presença de bioflme supragengival e gengivite, respectivamente. Os colutórios contendo o óleo essencial das folhas de L. sidoides (1%) e o extrato das folhas de A. indica (25%) podem ser indicados como tratamento adjuvante da gengivite induzida por biofilme.