Antimicrobial activity of fractions and subfractions of Elaeagia utilis against microorganisms of importance in dental caries

La caries dental es una enfermedad infecciosa multifactorial que conduce a la destrucción del tejido duro dental. El principal objetivo de la investigación en plantas medicinales es la búsquedade compuestos con actividad antimicrobiana para su posterior uso en estrategias de prevención o control de enfermedades infecciosas. El objetivo de este estudio fue evaluar la actividad antimicrobiana de fracciones y subfracciones obtenidas de la planta Elaeagia utilis contra Streptococcus mutans, Streptococcus sobrinus y Lactobacillus acidophilus. El materialvegetal fue colectado en la ciudad de Albán (Cundinamarca-Colombia) situada a una altitud de 2245 metros sobre el nivel del mar. Mediante el método de maceración en frio de hojas de E. utilis se obtuvieron dos extractos, uno en éter de petróleo y otro en etanol. Del extracto etéreo se obtuvieron fraccionesmediante cromatografía en columna al vacío y al extracto etanólico se le realizó fraccionamiento líquido/líquido continuo. La evaluación de la actividad antimicrobiana de las fracciones ysubfracciones se realizó por el método de difusión en pozo. A una concentración de 10 mg/pozo, múltiples fracciones obtenidas de los dos extractos presentaron actividad antimicrobianasobre S. mutans, S. sobrinus y L. acidophilus. De las fracciones del extracto etanólico se destaca la fracción diclorometano, por presentar mayor actividad antimicrobiana, razón por lo cual se subfracciona y se obtienen tres subfracciones con actividad inhibitoria.La subfracción más activa fue eOH:H2O (Bp) con una concentración mínima inhibitoria de 0.1 mg/pozo sobre las 3bacterias en estudio. En esta subfracción se determinaron terpenos, sesquiterpenlactonas y compuestos fenólicos simples. En conclusión, en este estudio se presenta el potencial antimicrobiano de fracciones y subfracciones obtenidas de extractos dehojas de E. utilis contra microorganismos de importancia encaries dental.

Dental caries is a multifactorial infectious disease that leads tothe destruction of dental hard tissue. The main goal of researchinto medicinal plants is to seek compounds with antimicrobialactivity for subsequent use in prevention strategies and controlof infectious diseases. The aim of this study was to evaluate theantimicrobial activity of fractions and subfractions obtainedfrom Elaeagia utilis against Streptococcus mutans, Streptococ-cus sobrinus and Lactobacillus acidophilus. The plant materialwas collected in the town of Alban (Cundinamarca, Colombia),which is located at an altitude of 2245 meters above sea level.Two extracts were obtained by cold maceration of E. utilis leavesin (a) petroleum ether extract and (b) ethanol extract. Fractionswere obtained from the petroleum ether extract by column vacu-um chromatography, and from the ethanol extract by continuousliquid / liquid partitioning. The antimicrobial activity of frac-tions and subfractions was evaluated by the well diffusionmethod. At a concentration of 10 mg /well, several fractions fromboth extracts showed antimicrobial activity against S. mutans, S.sobrinus and L. acidophilus. Among the ethanol extract frac-tions, the dichloromethane fraction had notably greaterantimicrobial activity. It was sub-partitioned, yielding three sub-fractions with inhibitory activity, of which the most active wasMeOH: H2O (Bp) with minimum inhibitory concentration 0.1mg /well on the 3 study bacteria. Terpenes, sesquiterpenlactonesand simple phenolic compounds were identified in it. In conclu-sion, this study shows the antimicrobial potential of fractionsand subfractions obtained from extracts of E. utilis leaves againstbacteria that are important in dental caries.

Experiência de cárie dentária e presença de estreptococos do grupo mutans em indivíduos com fissuras labio-palatinas: estudo transversal controlado / Dental caries experience and presence of mutans streptococci in subjects with cleft lip and palate: controlled cross-sectional study

Este estudo transversal controlado comparou a experiência de cárie e a presença de estreptococos do grupo mutans em adolescentes e adultos jovens com fissura labial e/ou palatina (FL/P) com um grupo controle sem fissura. Verificou também a associação entre experiência de cárie dentária e variáveis biológicas e ambientais relacionadas à doença. Foram incluídos 30 pacientes com FL/P (grupo caso) e 30 sem FL/P (grupo controle), com idade entre 12 e 21 anos, pareados por gênero, idade, condições de vida e utilização de dispositivo ortodôntico. Foram coletadas informações sobre: renda familiar per capita, contato com fluoretos, hábitos de higiene, acesso ao tratamento odontológico, escolaridade materna e contato com sacarose. No exame clínico foram registrados os índices de placa visível, de sangramento gengival, CPOD e CPOS, e as superfícies com lesões ativas de cárie. Foi coletado biofilme supra-gengival a partir do qual foram realizados procedimentos para isolamento e identificação por PCR de S. mutans e de S. Sobrinus. Os dados foram analisados utilizando o programa estatístico SPSS Statistics Data Editor 17.0, com nível de significância de 5%. Os dados clínicos foram analisados pelo teste de McNemar para comparar as variáveis qualitativas dicotômicas, pelo teste t pareado para as variáveis quantitativas com distribuição normal e pelo teste de Wilcoxon para as quantitativas com distribuição não-normal. Realizou-se comparação entre os grupos para a prevalência dos estreptococcos do grupo mutans. A associação entre a experiência de cárie, e as variáveis independentes foi verificada por análise multivariada por regressão logística backward stepwise conditional. Não houve diferença significativa entre os grupos caso e controle para higiene bucal e contato com fluoretos. Os grupos diferiram significativamente quanto às variáveis: renda per capita (p=0,007), presença de cárie ativa (p=0,024), componente cariado do CPOS (p=0,040), índice de placa visível (p=0,000) e sangramento gengival (p=0,002). Foi observada presença de S. mutans em 83,34% das amostras de biofilme do grupo caso e em 86,67% das do grupo controle; o S. sobrinus estava presente em 6,66% das amostras de biofilme do grupo caso e em 10% do grupo controle. Não houve associação entre a presença de S. mutans ou de S. sobrinus e experiência de cárie. A presença de FL/P mostrou-se associada à cárie apenas quando esta foi medida pelo CPOS. O uso de aparelho ortodôntico mostrou associação positiva com o CPOD, CPOS, componente restaurado do CPOD e CPOS. Os indivíduos com FL/P mostraram maior experiência de cárie que os sem FL/P. Dentre os fatores estudados que podem interferir no processo da doença, os fatores ambientais mostraram-se mais importantes que os fatores biológicos na determinação da experiência de cárie.

This controlled cross-sectional study aimed to compare the caries experience and presence of mutans streptococci in adolescents and young adults with cleft lip and / or palate (CL/P) with a control group without clefts. The association between dental caries experience and biological and environmental factors was also assessed. The study included 30 patients with CL/P (case group) and 30 without CL/P (control group), aged between 12 and 21 years, matched by gender, age, living conditions and use of orthodontic device. Data of family income, contact with fluorides, hygiene, access to dental care, maternal education and contact with sucrose were collected. Visible plaque index, gingival bleeding, DMFT and DMFS, and surfaces with active caries lesions were recorded. Microorganisms were isolated from the biofilm collected to identification of S. mutans and S. Sobrinus by PCR. The data were analyzed using the statistical program SPSS 17.0 Data Editor, with a significance level of 5%. The clinical data were analyzed by the McNemar test to compare dichotomous qualitative variables or by the paired t test for quantitative variables with normal distribution or the Wilcoxon test for quantitative variables with non-normal distribution. The groups were compared for the prevalence of mutans streptococci. Backward stepwise conditional logistic regression test the association between caries experience and independent variables. There was no significant difference between case and control groups for oral hygiene and contact with fluoride. The groups differed significantly with respect to variables: per capita income (p = 0.007), surfaces with active caries (p = 0.024), the decay component of the DMFS index (p = 0.040), visible plaque index (p = 0.000) and gingival bleeding (p = 0.002). S. mutans was observed in 83.34% of the biofilm samples in the case group and 86.67% of the control group; S. sobrinus was present in 6.66% of the biofilm samples in the case group and 10% in the control group. There was no association between the presence of S. mutans or S. sobrinus and caries. The presence of CL/P was associated with caries only when it was measured by DMFS. The use of orthodontic apparatus was positively associated with the DMFT, DMFS, the filled components of DMFT and DMFS. Subjects with CL/P showed a higher caries experience than those without CL/P. Among the studied factors that may influence the caries process, environmental factors were more important than biological factors in determining disease.

Development of Species-Specific PCR Primers for the Detection of Streptococcus sobrinus

This study was undertaken to develop species-specific forward and universal reverse PCR primers for the detection of Streptococcus sobrinus. These primers target the variable regions of the 16S ribosomal RNA coding gene (rDNA) and their specificity was tested against 10 strains of S. sobrinus strains and 20 different species of oral bacteria using serial dilutions of the purified genomic DNA of S.sobrinus ATCC 33478T . Our data show that species-specific amplicons were obtained from all the S. sobrinus strains tested but not from other species. Both direct and nested PCR could detect as little as 400 pg and 4 fg of genomic DNA from S. sobrinus ATCC 33478T , respectively. This result suggests that these PCR primers are highly specific and sensitive and applicable to the detection of S. sobrinus.

Effect of propolis on cariogenesis of main cariogenic streptococci and their fluoride-resistant strains / 第三军医大学学报

Objective To evaluate the effects of propolis on the growth of S.mutans ATCC 25175(S.m),S.sobrinus 6715(S.s) and their fluoride-resistant strains(S.m-FR,S.s-FR),and the change of their glucosyltransferase(GTF).Methods S.m and S.s were induced with sodium fluoride by minimum inhibitory concentration(MIC) to induce fluoride-resistant strains(S.m-FR and S.s-FR).Fluid dilution method was used to observe the effects of different levels of propolis on growth of S.m,S.m-FR,S.s and S.s-FR.Chemistry enzyme analysis was used to detect the influence of propolis on the enzyme activity of GTF.Results MIC of propolis for S.m,S.m-FR,S.s and S.s-FR were respectively 0.39,0.78,0.20 and 0.39 g/L.MBC of propolis for them were respectively 0.78,1.56,1.56,and 1.56 g/L.GTF of S.m,S.m-FR,S.s,and S.s-FR was decreased gradually with the increase of concentration of propolis.There were significant differences among total sample groups,and each experimental group and positive control group as well(P

Streptococcus mutans isolates identified by colony morphology,biochemical tests and DNA G+C mol% / 实用口腔医学杂志

砄bjective:To investigate the consistency of Streptococcus mutans and Streptococcus sobrinus isolates identified by colony morphology,biochemical tests and DNA base content analysis.Methods:46 clinical isolates (30 of S.mutans and 16 of S.sobrinus ) were identified by colony morphology,biochemical tests and determination of DNA base contents with reversed phase high performance liquid chromatography. S.mutans Ingbritt and S.sobrinus 6715 were used as the controls. Results:The DNA contents (G+C mol%) of S.mutans and S.sobrinus clinical isolates, identified by colony morphdogy and biochemical test, were 37.52 and 45.19 respectively,consistent with the expected 36～38 and 44～46 mol percent of G+C.Conclusion: S.mutans and S.sobrinus isolates,identified by colony morphology and biochemical tests ,were consistent with the DNA G+C content analysis. S.mutans and S.sobrinus can be easily and reliably identified by colony morphology and biochemical tests.