Silvana Soléo Ferreira dos Santos

Effectiveness of Photodynamic Therapy Against Enterococcus faecalis, With and Without the Use of an Intracanal Optical Fiber: An In Vitro Study

BACKGROUND AND OBJECTIVE Photodynamic therapy (PDT) is a new technique introduced in endodontics that combines the action of a photosensitizer (dye) and a low intensity light source. Currently, there are no PDT studies evaluating the microbial disinfection of root canals in order to compare the effects of light delivery systems in the photosensitizer activation. The aim of this study was to evaluate the PDT effectiveness in reducing Enterococcus faecalis, with and without the aid of an intracanal optical fiber. METHODS Extracted single-rooted teeth were selected, instrumented, inoculated with E. faecalis and divided into six groups: one control group (untreated), one conventionally-treated group (1% NaOCl irrigation) and four PDT-treated groups. Irradiation (diode laser) was performed with (OF) or without an intracanal optical fiber (NOF) using two different irradiation times: 1 min and 30 sec (IT(90)) or 3 min (IT(180)). Samples were collected before and after testing procedures and CFU/mL was determined. RESULTS The greatest reduction of E. faecalis (99.99%) was achieved with irrigation with 1% NaOCl. PDT also significantly reduced E. faecalis in the following decreasing order: OF/IT(180), NOF/IT(180), OF/IT(90) and NOF/IT(90), with no significant statistical difference among the groups. CONCLUSIONS These results suggest that PDT was effective against E. faecalis, regardless of the use of an intracanal optical fiber.

Effects of probiotic bacteria on Candida presence and IgA anti-Candida in the oral cavity of elderly.

Imbalance in the resident microbiota may promote the growth of opportunistic microorganisms, such as yeasts of Candida genus and the development of diseases, especially in aged people. This study evaluated whether the consumption of the probiotic Yakult LBz(®) (Lactobacillus casei and Bifidobacterium breve) was able to influence on the specific immunological response against Candida and on the presence of these yeasts in the oral cavity of 42 healthy aged individuals. Saliva samples were collected before and after the probiotic use for 30 days, 3 times a week. The samples were plated in Dextrose Saboraud Agar with chloramphenicol, the colony-forming units (CFU/mL) were counted and the Candida species were identified. Anti-Candida IgA analysis was conducted using the ELISA technique. ANOVA and Students t-test were used for normally distributed data and the Wilcoxon test was used for data with non-normal distribution (α=0.05). The results showed a statistically significant reduction (p<0.05) in Candida prevalence (from 92.9% to 85.7%), in CFU/mL counts of Candida and in the number of non-albicans species after consumption of the probiotic. Immunological analysis demonstrated a significant increase (p<0.05) in anti-Candida IgA levels. In conclusion, probiotic bacteria reduced Candida numbers in the oral cavity of the elderly and increased specific secretory immune response against these yeasts, suggesting its possible use in controlling oral candidosis.

Stability of antimicrobial activity of peracetic acid solutions used in the final disinfection process

The instruments and materials used in health establishments are frequently exposed to microorganism contamination, and chemical products are used before sterilization to reduce occupational infection. We evaluated the antimicrobial effectiveness, physical stability, and corrosiveness of two commercial formulations of peracetic acid on experimentally contaminated specimens. Stainless steel specimens were contaminated with Staphylococcus aureus, Escherichia coli, Candida albicans, blood, and saliva and then immersed in a ready peracetic acid solution: 2% Sekusept Aktiv (SA) or 0.25% Proxitane Alpha (PA), for different times. Then, washes of these instruments were plated in culture medium and colony-forming units counted. This procedure was repeated six times per day over 24 non-consecutive days. The corrosion capacity was assessed with the mass loss test, and the concentration of peracetic acid and pH of the solutions were measured with indicator tapes. Both SA and PA significantly eliminated microorganisms; however, the SA solution was stable for only 4 days, whereas PA remained stable throughout the experiment. The concentration of peracetic acid in the SA solutions decreased over time until the chemical was undetectable, although the pH remained at 5. The PA solution had a concentration of 500-400 mg/L and a pH of 2-3. Neither formulation induced corrosion and both reduced the number of microorganisms (p = 0.0001). However, the differences observed in the performance of each product highlight the necessity of establishing a protocol for optimizing the use of each one.

Antimicrobial activity of Uncaria tomentosa against oral human pathogens

Uncaria tomentosa is considered a medicinal plant used over centuries by the peruvian population as an alternative treatment for several diseases. Many microorganisms usually inhabit the human oral cavity and under certain conditions can become etiologic agents of diseases. The aim of the present study was to evaluate the antimicrobial activity of different concentrations of Uncaria tomentosa on different strains of microorganisms isolated from the human oral cavity. Micropulverized Uncaria tomentosa was tested in vitro to determine the minimum inhibitory concentration (MIC) on selected microbial strains. The tested strains were oral clinical isolates of Streptococcus mutans, Staphylococcus spp., Candida albicans, Enterobacteriaceae and Pseudomonas aeruginosa. The tested concentrations of Uncaria tomentosa ranged from 0.25-5% in Müeller-Hinton agar. Three percent Uncaria tomentosa inhibited 8% of Enterobacteriaceae isolates, 52% of S. mutans and 96% of Staphylococcus spp. The tested concentrations did not present inhibitory effect on P. aeruginosa and C. albicans. It could be concluded that micropulverized Uncaria tomentosa presented antimicrobial activity on Enterobacteriaceae, S. mutans and Staphylococcus spp. isolates.

Staphylococcus spp. in the oral cavity and periodontal pockets of chronic periodontitis patients

Staphylococcus spp. nao sao usualmente isolados a partir da cavidade bucal. Quando presentes, sao considerados pertencentes a microbiota transitoria. Individuos que apresentam doenca periodontal representam possiveis reservatorios dessas bacterias oportunistas na cavidade bucal. O uso de antibioticos para o tratamento da doenca periodontal ou outras infeccoes pode predispor o aumento do numero de Staphylococcus spp. na boca, pois estes adquirem facilmente resistencia aos antibioticos, podendo resultar em superinfeccao. O objetivo deste estudo foi verificar a presenca de Staphylococcus spp. na cavidade bucal e nas bolsas periodontais de pacientes com periodontite cronica; identificar as cepas isoladas; verificar a relacao entre a presenca de Staphylococcus spp. na cavidade bucal e presenca de bolsa periodontal. Participaram deste estudo 88 pacientes, entre 25 e 60 anos de idade e apresentando periodontite cronica, com pelo menos dois sitios com profundidade de sondagem maior ou igual a 5mm. Apos anamnese e exame clinico periodontal foram feitas coletas de material da bolsa periodontal com cones de papel e da cavidade bucal por meio de bochechos. Do total de pacientes 37,50% apresentaram Staphylococcus spp. na bolsa periodontal e 61,36% na cavidade bucal, sendo que 27,27% apresentaram a bacteria nos 2 sitios. S. epidermidis foi a especie mais prevalente para bolsa periodontal (15,9%) e cavidade bucal (27,27%). Nao houve diferenca estatistica significante quanto a presenca desses microrganismos entre as faixas etarias e aumento da profundidade de sondagem. A presenca de bacterias oportunistas na cavidade bucal pode representar dificuldades para a manutencao do tratamento periodontal.

Influence of probiotics on Candida presence and IgA anti-Candida in the oral cavity

Probiotics are defined as microorganisms that promote benefits to host health, mainly by regulating resident microbiota. Disequilibrium in microbiota can favor the growth of opportunist microorganisms and the development of pathologies, like candidosis caused by yeasts of the Candida genus. This work evaluated whether probiotics consumption was able to influence a specific immunological response to Candida and the presence of these yeasts in the oral cavity. Saliva samples were collected from healthy individuals and plated in Dextrose Saboraud Agar with chloramphenicol. Individuals presenting Candida in the oral cavity used the probiotic Yakult LB® for 20 days, after which new collections and identifications were performed. Anti-Candida IgA analysis was conducted using the ELISA technique. Analysis of the results showed a significant reduction in Candida prevalence (46%) and mean Candida CFU/mL counts (65%). The Candida species identified were C. albicans (98%) and C. tropicalis (2%), before and after probiotics consumption. Immunological analysis demonstrated a significant reduction in anti-Candida IgA levels after probiotics use, probably due to less antigenic stimulation. In conclusion, in the individuals studied, probiotics use significantly reduced the amount of Candida in the oral cavity, possibly due to competition between the yeasts rather than by specific secretory immune response stimulation.

Effectiveness of three instrumentation systems to remove Enterococcus faecalis from root canals

AIM To assess the effectiveness of three systems of mechanical preparation to reduce Enterococcus faecalis within root canals. METHODOLOGY Twenty-four human single-rooted canine teeth were standardized to a length of 17 mm and the canal contents removed using a size 20 K-file, as the last apical file. After irrigation and sterilization, the canals were contaminated with E. faecalis and incubated for 21 days at 37 °C with 5% CO(2). Then, the teeth were divided into three groups for mechanical preparation with: ProTaper rotary system, ProTaper manual system and manual K-files. Samples of the root canal contents, before and after the debridement, were collected with sterile paper points for 1 min. Then, the samples were diluted and plated in Brain Heart Infusion (BHI) agar. The colony-forming units were counted and the percentage reduction calculated. The reduction and log CFU mL(-1) were compared between groups using Wilcoxon nonparametric test and two-way analysis of variance, respectively. RESULTS There was a significant reduction in the number of CFU/mL (P = 0.000) before and after debridement for all the systems used. However, there was no significant difference between the systems. CONCLUSION All the three instrumentation systems reduced E. faecalis counts to a similar degree.

Staphylococcus spp., Enterobacteriaceae and Pseudomonadaceae oral isolates from Brazilian HIV-positive patients. Correlation with CD4 cell counts and viral load

The aim was to evaluate the presence of Staphylococcus spp., Enterobacteriaceae and Pseudomonadaceae in the oral cavities of HIV-positive patients. Forty-five individuals diagnosed as HIV-positive by ELISA and Western-blot, and under anti-retroviral therapy for at least 1 year, were included in the study. The control group constituted 45 systemically healthy individuals matched to the HIV patients to gender, age and oral conditions. Oral rinses were collected and isolates were identified by API system. Counts of microorganisms from HIV and control groups were compared statistically by a Mann-Whitney test (α=5%). The percentages of individuals positive for staphylococci were similar between the groups (p=0.764), whereas for Gram-negative rods, a higher percentage was observed amongst HIV-positive (p=0.001). There was no difference in Staphylococcus counts between HIV and control groups (p=0.1008). Counts were lower in the oral cavities of patients with low viral load (p=0.021), and no difference was observed in relation to CD4 counts (p=0.929). Staphylococcus aureus was the most frequently isolated species in HIV group, and Staphylococcus epidermidis was the prevalent species in the control group. Significantly higher numbers of enteric bacteria and pseudomonas were detected in the oral cavities of the HIV group than in the control (p=0.0001). Enterobacter cloacae was the most frequently isolated species in both groups. Counts of enteric bacteria and pseudomonas were significantly lower in patients with low CD4 counts (p=0.011); however, there was no difference relating to viral load. It may be concluded that HIV group showed greater species diversity and a higher prevalence of Enterobacteriaceae/Pseudomonadaceae.

Enterobacteriaceae and pseudomonadaceae on the dorsum of the human tongue

Objective: The aim of this study was to correlate the presence of Enterobacteriaceae, Pseudomonadaceae, Moraxellaceae and Xanthomonadaceae on the posterior dorsum of the human tongue with the presence of tongue coating, gender, age, smoking habit and denture use. Material and Methods: Bacteria were isolated from the posterior tongue dorsum of 100 individuals in MacConkey agar medium and were identified by the API 20E system (Biolab-Mérieux). Results: 43% of the individuals, presented the target microorganisms on the tongue dorsum, with greater prevalence among individuals between 40 and 50 years of age (p = 0.001) and non-smokers (p=0.0485). Conclusions: A higher prevalence of Enterobacteriaceae and Pseudomonadaceae was observed on the tongue dorsum of the individuals evaluated. There was no correlation between these species and the presence and thickness of tongue coating, gender and presence of dentures.

In vitro minocycline activity on superinfecting microorganisms isolated from chronic periodontitis patients

Chronic periodontitis is the most common type of periodontitis and it is associated with various species of microorganisms. Enteric rods, Pseudomonas, Staphyloccocus and Candida have been retrieved from periodontal pockets of patients with chronic periodontitis and correlated to cases of superinfection. Local or systemic antibiotic therapy is indicated to reinforce the effects of the conventional mechanical therapy. Minocycline has been suggested as one of the most effective drugs against periodontal pathogens. The aim of this work was to evaluate the minimal inhibitory concentration (MIC) of minocycline on superinfecting microorganisms isolated from the periodontal pocket and the oral cavity of individuals with chronic periodontitis. Isolates of Enterobacteriaceae (n = 25), Staphylococcus spp. (n = 25), Pseudomonas aeruginosa (n = 9) and Candida spp. (n = 25) were included in the study. Minimal inhibitory concentrations (MIC) of minocycline were determined using the Müeller-Hinton agar dilution method. Staphylococcus spp. isolates were the most sensitive to minocycline with a MIC of 8 microg/mL, followed by Enterobacteriaceae with a MIC of 16 microg/mL. The concentration of 16 microg/mL inhibited 96% of Candida spp. isolates. The MIC for 88.8% of the isolates of Pseudomonas aeruginosa was 128 microg/mL. A concentration of 1,000 microg/mL was not enough to inhibit 100% of the tested isolates.