Efeito de extratos vegetais de Hamamelis virginiana, Persea americana, Cynara scolymus L e Stryphnodendron barbatiman Msobre a expressão fenotípica dos fatores de virulência em biofilmes de Candida albicans / Effect of Hamamelis virginiana, Persea americana, Cynara scolymus L and Stryphnodendron barbatiman M plant extracts on the phenotypic expression of virulence factors in biofilms of the Candida albicans

Objetivo: Analisar a expressão fenotípica de fatores de virulência em biofilmes de Candida albicans frente a extratos glicólicos de plantas. Material e Métodos: Os biofilmes de Candida albicans (ATCC 18804) obtidos a partir de incubação de 48 horas foram expostos por 5 minutos e 24 horas a diferentes concentrações de extratos glicólicos de Hamamelis virginiana e Persea americana, Cynara scolymus L e Stryphnodendron barbatiman M, a fim de verificar a ação antifúngica da proteinase, fosfolipase e hemolisina. Resultados: Todos os extratos foram eficazes na redução do biofilme. Em contato por 5 minutos. os extratos reduziram 50% do biofilme. Após 24 horas. o extrato de Persea americana apresentou o biofilme em 90%, seguido de Cynara scolymus, que o interrompeu em 85%. Houve mudança na intensidade da proteinase após 5 minutos e 24 horas, com uma atividade enzimática média de 0,69 em comparação com o controle de 0,49. Cynara scolymus foi o extrato com maior concentração média de 100 mg/ml; a intensidade da fosfolipase foi alterada com Stryphnodendron barbatiman sendo mais efetivo em 24 horas em relação ao controle (p< 0,0001). A secreção de hemolisina foi modificada por Hamamelis virginiana (12,5 mg/ml) após 5 minutos de exposição e em 24 horas. todos os extratos foram capazes de causar alterações na secreção. Conclusão: Os extratos testados apresentam potencial antifúngico em biofilmes de Candida albicans, implicando em redução significativa dos fatores de virulência. Assim, estes podem ser indicados como uma ferramenta terapêutica alternativa para reduzir a morbidade dessas infecções, já que em ambos os tempos de exposição investigados, eles foram capazes de reduzir a secreção enzimática do fungo (AU)

Objective: Analyze the phenotypic expression of virulence factors in Candida albicans biofilms against plant glycolicextracts. Material and Methods: The biofilms of Candida albicans (ATCC 18804) obtained from incubation for 48 hours were exposed for 5 minutes and 24 hours to different concentrations of glycolic extracts of Hamamelis virginiana and Persea americana, Cynara scolymus L and Stryphnodendron barbatiman M, in order to verify the antifungal activity of the proteinase, phospholipase and hemolysin. Results: All extracts were effective in reducing biofilm. In contact for 5 minutes. the extracts reduced 50% of the biofilm. After 24 hours, the Persea americanaextract showed the biofilm at 90%, followed by Cynara scolymus, which interrupted it at 85%, There was a change in proteinase intensity after 5 minutes and 24 hours. with an average enzymatic activity of 0.69 compared to the control of 0.49. Cynara scolymus was the extract with the highest mean concentration of 100 mg/ml; the phospholipase intensity was changed with Stryphnodendron barbatiman being more effective in 24 hours compared to the control (p< 0.0001). The hemolysin secretion was modified by Hamamelis virginiana (12.5 mg/ml) after 5 minutes of exposure, and in 24 hours. all extracts were capable to cause changes in secretion. Conclusion: The tested extracts have antifungal potential in Candida albicans biofilms, implying a significant reduction in virulence factors. Thus, these can be indicated as an alternative therapeutic tool to reduce the morbidity of these infections, as in both investigated exposure times. they were able to reduce theenzymatic secretion of the fungus (AU)

Candida albicans Mrv8, is involved in epithelial damage and biofilm formation.

Candida albicans is the most common human fungal pathogen that can cause superficial and deep-seated infections in susceptible individuals. Despite its medical importance, the vast majority of C. albicans genes remain of unknown function. Here, we report a role for the lineage-specific gene, MRV8, in host pathogen interactions, mycelial microcolony maturation and biofilm formation. In silico analysis indicated that MRV8 encodes a four-pass transmembrane protein unique to the closely related pathogens C. albicans and Candida dubliniensis. Deletion of MRV8 did not affect C. albicans adherence to, or initial invasion into human oral epithelia, but inhibited mycelial development and strongly reduced epithelial damage. mrv8Δ/Δ cells exhibited a media-dependent defect in biofilm formation and mutant biofilm metabolic activity was enhanced by cyclosporin A. mrv8Δ/Δ biofilms were more tolerant to treatment with caspofungin, but not to fluconazole or amphotericin B. Co-stimulation with calcium chloride and calcofluor white rescued biofilm growth in the presence of caspofungin, and this rescue-effect was Mrv8-dependent. Together, our data demonstrate an important role for a lineage-specific gene (MRV8) in C. albicans biofilm formation, drug tolerance and host-pathogen interactions.

Biofilms of Candida albicans and Streptococcus sanguinis and their susceptibility to antimicrobial effects of photodynamic inactivation.

This study evaluated the effects of photodynamic inactivation (PDI) on single and multi-species biofilms, compounds by Candida albicans and Streptococcus sanguinis. Biofilms were formed, on microplate of 96 wells, by suspensions of C. albicans (ATCC 18804) and S. sanguinis (ATCC 7073) adjusted in 107 cells/mL, followed by incubation of 48 h (with 5% CO2). The effects of the photosensitizer erythrosine (ER) at 400 µM for 5 min and green light-emitting diode (LED - 532 ±â€¯10 nm) for 3 min, alone and conjugated, were evaluated. After normality test, results was analysed by Tukey´s test (P < 0.05). PDI group promoted reductions of 1.07 and 0.39 log10, respectively, in biofilms of C. albicans alone and in association with S. sanguinis. Biofilms of S. sanguinis alone were more sensitive, with reduction of 4.48 log10. When in association with the yeast, S. sanguinis have a decrease of 2.67 log10. SEM analysis revealed a decrease in bacterial and fungal structures of biofilms treated with PDI. In conclusion PDI promoted significant microbial reductions in both species of microorganisms grown on mixed biofilms. This study is one of the pioneers to evaluate the antimicrobial action of PDI on biofilms of S. sanguinis and C. albicans, demonstrating a way to control these microorganisms of clinical importance.

Effect of ozonized olive oil on oral levels of Candida spp. in patients with denture stomatitis / Efeito do óleo de oliva ozonizado sobre os níveis orais de Candida spp. em pacientes com estomatite protética

Objective: The aim of this study was to evaluate the effect of ozonized olive oil (OZ) on the oral levels of Candida spp. in patients with denture stomatitis. Material and Methods: In vitro tests were performed to validate antifungal activity and to standardize OZ conditions. Antifungal activity was screened against C. albicans and five non-albicans species (C. tropicalis, C. dubliniensis, C. krusei, C. guilliermondii, and C. parapsilosis). Also, the effects on C. albicans planktonic and biofilm were evaluated. After validation, OZ was included in a therapeutic protocol of denture stomatitis in vivo. Thirty patients used OZ and 20 used sodium bicarbonate (SB) for 14 days. After 7 and 14 days, clinical evaluation, isolation and identification of yeasts were performed. Isolates were identified by phenotypic and genotypic tests. Ozonated oil showed in vitro antifungal activity against all species of Candida. Ozonated oil reduced the number of viable cells in C. albicans biofilms. Oral candidal levels were lower in relation to baseline both after after 14 days of treatment with SB and OZ. Results: A total of 493 Candida spp. isolates was obtained and 80% were identified as C. albicans. Remission of denture stomatitis was observed in all patients after 7 days of treatment in both groups. Conclusion: Within the limits of the study we can conclude that ozonized olive oil can be a new alternative for the control of biofilm in patients with denture stomatitis. (AU)

Objetivo: O objetivo deste estudo foi avaliar o efeito do óleo ozonizado (OZ) sobre os níveis orais de Candida spp. em pacientes com estomatite protética. Material e Métodos: Testes in vitro foram realizados para validar a atividade antifúngica e padronizar as condições do OZ. A atividade antifúngica foi avaliada contra C. albicans e cinco espécies não-albicans (C. tropicalis, C. dubliniensis, C. krusei, C. guilliermondii e C. parapsilosis). Além disso, os efeitos sobre C. albicans planctônico e biofilme foram avaliados. Após validação, o OZ foi incluído em um protocolo terapêutico de estomatite protética in vivo. Trinta pacientes usaram OZ e 20 usaram bicarbonato de sódio (SB) por 14 dias. Após 7 e 14 dias, foram realizadas avaliações clínicas, isolamento e identificação de leveduras. Os isolados foram identificados por testes fenotípicos e genotípicos. O óleo ozonizado mostrou atividade antifúngica in vitro contra todas as espécies de Candida. O óleo ozonizado reduziu o número de células viáveis em biofilmes de C. albicans. Os níveis orais de candidíase foram menores em relação aos valores basais após 14 dias de tratamento com SB e OZ. Resultados: Um total de 493 Candida spp. isolados foram obtidos e 80% foram identificados como C. albicans. A remissão da estomatite protética foi observada em todos os pacientes após 7 dias de tratamento em ambos os grupos. Conclusão: Dentro dos limites do estudo podemos concluir que o óleo de oliva ozonizado pode ser uma nova alternativa para o controle do biofilme em pacientes com estomatite protética.(AU)

Production of virulence factors in Candida strains isolated from patients with denture stomatitis and control individuals.

The aim of this study was to evaluate the production of virulence factors in Candida isolates from the oral cavities of 50 patients with different degrees of denture stomatitis (DS, type I, II and III) and 50 individuals without signs of DS. We evaluated the enzymatic and hemolytic activities, the biofilm formation, and the cell surface hydrophobicity (CSH) in all isolates. Germ tube (GT) production was also evaluated in Candida albicans and Candida dubliniensis isolates. In C. albicans and C. dubliniensis the secretion of hemolysin and GT production was significantly different between isolates from patients with DS and individuals without DS. No significant difference was observed in the production of virulence factors by Candida glabrata isolates. Candida isolates expressed a wide range of virulence factors. However, in the majority of isolates from the type III lesions, the production of the virulence factors was higher than for the other groups.

Oral microbial colonization in children with sickle cell anaemia under long-term prophylaxis with penicillin.

BACKGROUND AND OBJECTIVE: Sickle cell anaemia (SCA) is the most frequent haematological hereditary disease. Children with SCA are submitted to long-term prophylactic therapy with penicillin, but little is known about its impact on oral microflora. The aim of this study was to evaluate the oral microbial colonization of paediatric patients with SCA. DESIGN: Forty children (4-11yrs old) with SCA (genotype SS) under long-term prophylactic treatment with penicillin were included in the study. Age/gender-matched control group of healthy children was also included. Scores of dmft/DMFT (number of decayed (D), missing (M), or filled (F) teeth; dmft, for primary dentition; DMFT, for permanent dentition) were obtained and stimulated saliva was sampled. Salivary flow rate and buffering capacity were evaluated. Counts of microorganisms (mutans streptococci, lactobacilli and yeasts) were determined by plating method. Yeasts were identified by API 20C AUX and PCR. RESULTS: Mean dmft/DMFT values were similar in the studied groups (SCA 2.13/1.60 and control 2.38/1.3). Although no significant differences between cariogenic microorganism counts were observed, significantly higher yeasts oral levels were observed in SCA group. Controls showed lower salivary buffering capacity. Candida albicans was the most frequently isolated species in both groups. Candida famata, Candida parapsilosis and Candida tropicalis were also isolated from controls. Candida dubliniensis, Candida rugosa and Candida sphaerica were found only in SCA group. CONCLUSIONS: Based on the results, it could be concluded that paediatric patients with SCA showed significantly higher oral level of yeasts. Uncommon fungal species were found in SCA group. Similar caries prevalence and counts of lactobacilli and streptococci in relation to controls were observed.

Opportunistic microorganisms in individuals with lesions of denture stomatitis.

The aim of this study was to isolate, quantify, identify, and compare opportunistic microorganisms (Candida and Staphylococcus genera and Enterobacteriaceae/Pseudomonadaceae families) from prosthesis-fitting surfaces, the hard palate, and mouth rinses of individuals wearing removable maxillary prosthesis with (50) and without (50) lesions of denture stomatitis (DS). The strains were collected and identified using phenotypic, biochemical and molecular tests. The counts of microorganisms were significantly higher in the group of individuals with DS (P < 0.05). C. albicans was the most frequently isolated yeast species in both groups, following by C. tropicalis and C. glabrata. Six isolates were identified as C. dubliniensis. S. aureus and S. epidermidis were the most frequent Staphylococcus species in both groups. Klebsiella pneumoniae was the predominant species in both groups. The association between Candida spp. and bacteria isolated in this study with DS suggests that these microorganisms may play important roles in the establishment and persistence of this disease.

Effects of eating disorders on oral fungal diversity.

BACKGROUND: The eating disorders anorexia and bulimia nervosa can cause several systemic and oral alterations related to poor nutrition and induced vomiting; however, the oral microflora of these patients is poorly studied. OBJECTIVE: The aim of this study was to evaluate fungal microflora in the oral cavity of these patients by culture-dependent and culture-independent methods. STUDY DESIGN: Oral rinse samples were cultured to assess the prevalence of Candida species, and the isolates were identified by API system. Microorganism counts were compared by the Mann-Whitney test (5%). Ribotyping, a type of molecular analysis, was performed by sequencing the D1/D2 regions of 28S rRNA. RESULTS: Our results demonstrated that the eating disorder group showed higher oral Candida spp. prevalence with culture-dependent methods and higher species diversity with culture-independent methods. CONCLUSIONS: Eating disorders can lead to an increased oral Candida carriage. Culture-independent identification found greater fungal diversity than culture-dependent methods.

Molecular identification of Pichia guilliermondii, Debaryomyces hansenii and Candida palmioleophila.

Traditional phenotypic methods and commercial kits based on carbohydrate assimilation patterns are unable to consistently distinguish among isolates of Pichia guilliermondii, Debaryomyces hansenii and Candida palmioleophila. As result, these species are often misidentified. In this work, we established a reliable method for the identification/differentiation of these species. Our assay was validated by DNA sequencing of the polymorphic region used in a real-time PCR assay driven by species-specific probes targeted to the fungal ITS 1 region. This assay provides a new tool for pathogen identification and for epidemiological, drug resistance and virulence studies of these organisms.

Molecular identification of Pichia guilliermondii, Debaryomyces hansenii and Candida palmioleophila

Traditional phenotypic methods and commercial kits based on carbohydrate assimilation patterns are unable to consistently distinguish among isolates of Pichia guilliermondii, Debaryomyces hansenii and Candida palmioleophila. As result, these species are often misidentified. In this work, we established a reliable method for the identification/differentiation of these species. Our assay was validated by DNA sequencing of the polymorphic region used in a real-time PCR assay driven by species-specific probes targeted to the fungal ITS 1 region. This assay provides a new tool for pathogen identification and for epidemiological, drug resistance and virulence studies of these organisms.

Opportunistic microorganisms in patients undergoing antibiotic therapy for pulmonary tuberculosis

Antimicrobial therapy may cause changes in the resident oral microbiota, with the increase of opportunistic pathogens. The aim of this study was to compare the prevalence of Candida, Staphylococcus, Pseudomonas and Enterobacteriaceae in the oral cavity of fifty patients undergoing antibiotic therapy for pulmonary tuberculosis and systemically healthy controls. Oral rinsing and subgingival samples were obtained, plated in Sabouraud dextrose agar with chloramphenicol, mannitol agar and MacConkey agar, and incubated for 48 h at 37ºC. Candida spp. and coagulase-positive staphylococci were identified by phenotypic tests, C. dubliniensis, by multiplex PCR, and coagulase-negative staphylococci, Enterobacteriaceae and Pseudomonas spp., by the API systems. The number of Candida spp. was significantly higher in tuberculosis patients, and C. albicans was the most prevalent specie. No significant differences in the prevalence of other microorganisms were observed. In conclusion, the antimicrobial therapy for pulmonary tuberculosis induced significant increase only in the amounts of Candida spp.

HAART therapy does not reduce the proteinase and phospholipase secretion by oral Candida albicans isolated from HIV-positive patients

Production of exoenzymes, specifically the proteinase and phospholipase, is considered one of the most important of pathogenicity mechanisms of C. albicans, which is crucial for tissue invasion. This study aimed at evaluating the production of these exoenzymes in 50 oral C. albicans isolates from HIV-positive (HIV+) patients treated with highly active anti-retroviral therapy (HAART), and from 50 control individuals. For testing the production of phospholipase and proteinase, the culture media containing egg yolk and bovine albumin were used, respectively. The results were obtained by measuring the diameter of the colony and divided by the diameter of colony plus the precipitation zone, defined as Pz. Data were statistically analyzed by Student’s t test (5). Statistically significant difference (p = 0.001) was observed between the mean values of Pz for proteinase in isolates from HIV+ patients (Pz = 0.358±0.295) and from control group (Pz = 0.660±0.370). The same results were observed for phospholipase production (Pz = 0.399±0.227 for HIV+ group; Pz =0.635±0.292 control group). Both enzymes were highly produced by C. albicans isolated from HIV+ patients when compared with those secreted by C. albicans obtained from control group, suggesting that HAART did not reduce the secretion of these enzymes by this pathogenic fungus infecting HIV+ patients.

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.

Opportunistic microorganisms in patients undergoing antibiotic therapy for pulmonary tuberculosis.

Antimicrobial therapy may cause changes in the resident oral microbiota, with the increase of opportunistic pathogens. The aim of this study was to compare the prevalence of Candida, Staphylococcus, Pseudomonas and Enterobacteriaceae in the oral cavity of fifty patients undergoing antibiotic therapy for pulmonary tuberculosis and systemically healthy controls. Oral rinsing and subgingival samples were obtained, plated in Sabouraud dextrose agar with chloramphenicol, mannitol agar and MacConkey agar, and incubated for 48 h at 37°C. Candida spp. and coagulase-positive staphylococci were identified by phenotypic tests, C. dubliniensis, by multiplex PCR, and coagulase-negative staphylococci, Enterobacteriaceae and Pseudomonas spp., by the API systems. The number of Candida spp. was significantly higher in tuberculosis patients, and C. albicans was the most prevalent specie. No significant differences in the prevalence of other microorganisms were observed. In conclusion, the antimicrobial therapy for pulmonary tuberculosis induced significant increase only in the amounts of Candida spp.

Evaluation of alternative methods for the disinfection of toothbrushes.

The aim of this study was to evaluate alternative methods for the disinfection of toothbrushes considering that most of the previously proposed methods are expensive and cannot be easily implemented. Two-hundred toothbrushes with standardized dimensions and bristles were included in the study. The toothbrushes were divided into 20 experimental groups (n = 10), according to microorganism considered and chemical agent used. The toothbrushes were contaminated in vitro by standardized suspensions of Streptococcus mutans, Streptococcus pyogenes, Staphylococcus aureus or Candida albicans. The following disinfectants were tested: 0.12% chlorhexidine digluconate, 50% white vinegar, a triclosan-containing dentifrice solution, and a perborate-based tablet solution. The disinfection method was immersion in the disinfectant for 10 min. After the disinfection procedure, the number of remaining microbial cells was evaluated. The values of cfu/toothbrush of each group of microorganism after disinfection were compared by Kruskal-Wallis ANOVA and Dunn's test for multiple comparisons (5%). The chlorhexidine digluconate solution was the most effective disinfectant. The triclosan-based dentifrice solution promoted a significant reduction of all microorganisms' counts in relation to the control group. As to the disinfection with 50% vinegar, a significant reduction was observed for all the microorganisms, except for C. albicans. The sodium perborate solution was the less effective against the tested microorganisms. Solutions based on triclosan-containing dentifrice may be considered effective, nontoxic, cost-effective, and an easily applicable alternative for the disinfection of toothbrushes. The vinegar solution reduced the presence of S. aureus, S. mutans and S. pyogenes on toothbrushes.

Evaluation of alternative methods for the disinfection of toothbrushes

The aim of this study was to evaluate alternative methods for the disinfection of toothbrushes considering that most of the previously proposed methods are expensive and cannot be easily implemented. Two-hundred toothbrushes with standardized dimensions and bristles were included in the study. The toothbrushes were divided into 20 experimental groups (n = 10), according to microorganism considered and chemical agent used. The toothbrushes were contaminated in vitro by standardized suspensions of Streptococcus mutans, Streptococcus pyogenes, Staphylococcus aureus or Candida albicans. The following disinfectants were tested: 0.12 percent chlorhexidine digluconate, 50 percent white vinegar, a triclosan-containing dentifrice solution, and a perborate-based tablet solution. The disinfection method was immersion in the disinfectant for 10 min. After the disinfection procedure, the number of remaining microbial cells was evaluated. The values of cfu/toothbrush of each group of microorganism after disinfection were compared by Kruskal-Wallis ANOVA and Dunn's test for multiple comparisons (5 percent). The chlorhexidine digluconate solution was the most effective disinfectant. The triclosan-based dentifrice solution promoted a significant reduction of all microorganisms' counts in relation to the control group. As to the disinfection with 50 percent vinegar, a significant reduction was observed for all the microorganisms, except for C. albicans. The sodium perborate solution was the less effective against the tested microorganisms. Solutions based on triclosan-containing dentifrice may be considered effective, nontoxic, cost-effective, and an easily applicable alternative for the disinfection of toothbrushes. The vinegar solution reduced the presence of S. aureus, S. mutans and S. pyogenes on toothbrushes.

Oral distribution of Candida species and presence of oral lesions in Brazilian leprosy patients under multidrug therapy.

OBJECTIVE: The aim of this study was to evaluate the prevalence of Candida spp. and presence of oral lesions in Brazilian leprosy patients under multidrug therapy (MDT). METHODS: Thirty-eight individuals (18 males and 20 females, median age 53 years) clinically and microbiologically diagnosed as leprosy (lepromatous variant), and under MDT for at least 45 days were studied. The control group constituted by 38 healthy individuals (median age 53.5), matched to the test group in relation to age, gender and oral conditions. Oral rinses were collected and the Candida identification was performed by phenotypic tests. The existence of Candida dubliniensis among the isolates was analyzed using a validated multiplex PCR assay. Twenty-nine leprosy patients were examined intra-orally for the presence of lesions. Data were analyzed by z- and Mann-Whitney tests (alpha = 5%). RESULTS: Yeast carriage rate between leprosy patients (65.8%) and controls (47.4%) was similar (P = 0.099), and no significant difference between yeast counts was observed (P = 0.1004). Candida albicans was the most frequently isolated species in both groups. In the leprosy group, Candida tropicalis and Candida parapsilosis were also identified. In the control group, we additionally identified Candida tropicalis, Candida glabrata and Candida kefyr. Candida dubliniensis was not detected. No leprosy-related oral lesion was registered. CONCLUSION: Within the limits of the study, we concluded that Brazilian leprosy patients under MDT showed similar levels of carriage and Candida species distribution in relation to the controls.

Avaliação da microbiota bucal de pacientes com anorexia nervosa e bulimia nervosa / Evaluation of oral microbiota of patients with Anorexia Nervosa and Bulimia Nervosa

Os Transtornos alimentares (TA) como Anorexia Nervosa (AN) e Bulimia Nervosa (BN) são acompanhados de inúmeras alterações sistêmicas e bucais relacionadas ao comprometimento do estado nutricional e às práticas compensatórias inadequadas para o controle do peso. O objetivo deste estudo foi avaliar diversidade microbiológica existente na cavidade bucal de pacientes com estes transtornos, por meio de técnicas de cultivo e utilizando métodos moleculares independentes de cultivo. Foram incluídos no estudo 32 pacientes anoréxicos e 27 bulímicos, pareados com 59 indivíduos controle. Amostras de enxágüe bucal foram semeadas para a avaliação da prevalência de leveduras do gênero Candida, estafilococos, enterococos, estreptococcos do grupo mutans (EGM), lactobacilos, enterobactérias/pseudomonas. Espécies de Candida, estafilococos, enterococos, enterobactérias/pseudomonas foram identificadas pelo sistema API. Amostras de biofilme supragengival foram coletadas e utilizadas somente nos procedimentos moleculares. As contagens de microrganismos nos grupos foram comparadas por ANOVA/Mann-Whitney (5%). Houve diferença estatisticamente significantes (p<0,05) para as contagem de leveduras do gênero Candida, estafilococos, enterococos, EGM e lactobacilos entre o grupo TA e controle, mas não houve diferenças significativas para a prevalência de enterobactérias/pseudomonas (p=0,312). Pequena diferença entre os grupos foi observada na diversidade de espécies dos microrganismos estudados pelo método de cultivo. Avaliação molecular foi realizada pela ribotipagem por seqüenciamento do 16S rRNA bacteriano e regiões D1/D2 do 28S rRNA. Foram avaliados cerca de 3000 clones do grupo TA e 1500 clones do controle. Sessenta e duas espécies ou filotipos de bactérias foram detectados, sendo que 22 identificações foram encontrados somente no grupo de estudo, 6 apenas no grupo controle e 34 em ambos os grupos.

Eating disorders such as nervous Anorexia and Bulimia Nervosa have several clinical and oral alterations related to the nutritional state involvement and the inadequate compensatory practices for weight control. The aim of this study was to evaluate the microbial diversity in the oral cavity of patients with Anorexia Nervosa and Bulimia nervosa by cultivation techniques and cultivation independent molecular methods. The study included 32 patients and 27 bulimic anorexics, matched with 59 control subjects. Oral rinse samples were cultured to assess the prevalence of Candida species, staphylococci, enterococci, streptococci mutans (EGM), lactobacilli, Enterobacteriaceae / Pseudomonas. Candida species, staphylococci, enterococci, Enterobacteriaceae / Pseudomonaswere identified by API systems. Supragingival biofilm samples were collected and used only in molecular procedures. Counts of microorganisms in the groups were compared by ANOVA / Mann-Whitney (5%). There was a statistically significant (p <0.05) for the counting of yeasts, staphylococci, enterococci, and lactobacilli EGM between TA and control groups, but there were no significant differences inthe prevalence of Enterobacteriaceae / Pseudomonas (p = 0.312). Few differences between the groups were observed in the species diversity of organisms studied by the method of cultivation. Molecular analysis was performed by ribotyping by sequencing the 16S rRNA bacterial and D1/D2 regions of 28S rRNA. About 3000 clones of the TA group and 1500 clones of control were evaluated. Sixty-two species or filotypes of bacteria were detected, with 22 identifications were found only in the study group, only 6 in the control group and 34 in both groups. Microorganisms related to caries and periodontal diseases were found in both groups.