Enzyme production by Candida albicans and Candida dubliniensis in periodontal HIV-positive patients receiving and not receiving antiretroviral therapy / Producción enzimática de Candida albicans y Candida dubliniensis de pacientes con periodontitis VIH+ con y sin tratamiento antirretroviral

ABSTRACT Candida dubliniensis (Cd) and Candida albicans (Ca) are the most frequently isolated yeasts in HIV+ patients. Some of the enzymes produced by these yeasts are considered virulence factors since they contribute to pathogenicity of Candida spp. The aim of the present study was to compare production of enzymes such as phospholipase (Ph), proteinase (P), and hemolysin (H) by Cd and Ca strains isolated from periodontal HIV-positive patients receiving and not receiving highly active antiretroviral therapy (HAART). Subgingival biofilm samples were obtained using paper points, and a sample of oral mucosa was taken using a swab. Phenotypic and molecular methods were used to isolate 39 strains of Candida, including 25 strains of Cd and 14 strains of Ca, obtained from 33 periodontal pocket samples and 6 oral mucosa samples collected from 15 HIV+ patients (8 receiving and 7 not receiving HAART). Malt egg-yolk agar, albumin agar and blood agar were used to evaluate pH, P and H production respectively. The strains were inoculated in duplicate and incubated at 37 ºC. Colony and halo diameters were measured. A greater proportion of Ca was observed in patients not receiving HAART, and a higher proportion of Cd was observed in those under HAART, Chi2 p< 0.001. Phospholipase production was observed in 92.9% percent of isolated Ca strains but in none of the isolated Cd strains. Proteinase production was high in Ca and Cd strains isolated from patients not receiving HAART. Hemolysin production was observed in all the studied strains, though it was significantly higher (p=0.04) in Ca and Cd strains isolated from patients not receiving HAART. To sum up, the proportion of Candida dubliniensis strains was highest in the subgingival biofilm of patients receiving HAART, and Cd strains were found to express fewer virulence factors than Ca strains.

RESUMEN Las levaduras más aisladas en pacientes VIH+ son Candida dubliniensis (Cd) y Candida albicans (Ca). Algunas de sus enzimas constituyen factores de virulencia ya que favorecen la diseminación tisular. El objetivo fue comparar la producción de enzimas como fosfolipasa (F), proteinasa (P) y hemolisina (H) en cepas de Cd y Ca aisladas de pacientes VIH+ tratados y no tratados con antirretrovirales (TARGA). Se realizó la toma del biofilm de placa subgingival con conos de papel y la muestra de la mucosa bucal con hisopo. Se aislaron y tipificaron por métodos fenotípicos y moleculares 39 cepas: 25 de Cd y 14 Ca, obtenidas 33 de bolsas periodontales y 6 de mucosa bucal de 15 pacientes VIH+ (8 con y 7 sin tratamiento). Se utilizó agar malta con yema de huevo, agar albúmina y agar sangre para demostrar la producción de F, P y H, respectivamente. Se inocularon por duplicado e incubaron a 37°C. Se midieron los diámetros de las colonias y los de hidrólisis alrededor de las mismas. Se observó mayor proporción de Ca en los pacientes sin tratamiento y mayor proporción de Cd en los con tratamiento; Chi2 p< 0.001. El 92,9% de las Ca estudiadas, fueron productoras de fosfolipasa. En tanto que ninguna Cd produjo la enzima. En cuanto a la producción de proteinasa se observa una alta producción tanto en las cepas de Ca, como en las Cd aisladas en los pacientes no tratados. Todas las cepas estudiadas produjeron hemolisina, observándose una diferencia estadísticamente significativa (p=0,04) en ambas especies a favor de la alta producción de la enzima en las cepas obtenidas de pacientes no tratados. Podemos concluir que en el biofilm subgingival, en los pacientes bajo TARGA, se aíslan mayor proporción de Candida dubliniensis las cuales expresan menos factores de virulencia.

Assessment of the oral microbiome in the development of recurrent oral aphthae in patients with gastrointestinal pathology

Chronic recurrent oral aphthae in residents living in an ecologically unfavourable region are characterized by a permanent course and prolonged recovery processes of regeneration of pathological elements of the oral mucosa. Using the microbiological method and modern test systems, it has been found that on the surface of aphthae an extremely diverse state of the oral microbiota is determined and its types are diverse. Trigger mechanisms have been determined. The role of representatives of various types of microorganisms - enterococci, staphylococci, streptococci, yeast-like fungi of the genus Candida (C. albicans) and obligate-anaerobes in the development of recurrent oral aphthae has been established. The data obtained can serve as an indication for the development of modern treatment and preventive measures regarding this category of patients.

Detection of intracellular Helicobacter pylori in Candida. SPP from neonate oral swabs

SUMMARY BACKGROUND: There is evidence of detection of Helicobacter pylori (H. pylori) in the stool of newborns and in the yeast that colonizes the oral cavity of this age group. However, there is a lack of research to confirm it. This study proposes to determine the existence of the bacteria at an early age, specifically in newborns. OBJECTIVE: To identify intracellular H. pylori in oral yeasts and to detect antigens of the bacteria in newborn stools. METHODOLOGY: Cross-sectional and descriptive study. Samples were obtained from infants (oral swab and meconium). Identification of yeast species was performed using the following techniques: CHROMagar Candida, Germinal Tube Test and API Candida Identification System, then the yeasts were observed by light microscopy and fluorescence. Detection of H. pylori antigen in meconium and PCR were performed to amplify specific genes of the bacterium (rRNA16S, cagA, vacA s1a, vacA s1b, vacA s2, vacA m1, vacA m2 and dupA). RESULTS: Intracellular H. pylori was detected in yeast of the species Candida glabrata (C. glabrata) isolated from an oral swab of a newborn. CONCLUSION: The results of this study evidenced the existence of intracellular H. pylori in newborns.

RESUMO ANTECEDENTES: Há evidências de detecçâo de Helicobacter pylori (H. pylori) em fezes de recém-nascidos, como também dentro de leveduras que colonizam a cavidade oral dessa faixa etária. No entanto, faltam investigações que confirmem esses achados. OBJETIVO: Identificar H. pylori intracelular em leveduras de origem oral e detectar antígenos dessa bactéria em fezes neonatais. METODOLOGIA: Estudo transversal e descritivo. As amostras foram obtidas de bebês (zaragatoa oral e mecônio). As identificações das espécies de leveduras foram realizadas utilizando as seguintes técnicas: CHROMagar Candida, teste de tubo germinativo e sistema de identificação API Cândida. As leveduras foram observadas por microscopía óptica e fluorescência. Realizou-se a detecçâo de antígeno de H. pylori em mecônio e PCR para a amplificação de genes específicos desta bactéria (rRNA16S, cagA, vacA s1a, vacA s1b, vacA s2, vacA m1, vacA m2 e dupA). RESULTADOS: Foi detectado H. pylori intracelular em leveduras da espécie Candida glabrata (C. glabrata) isoladas a partir de zaragatoas oral de um recém-nascido. CONCLUSÃO: Os resultados deste estudo evidenciaram a existência interna de levedura de H. pylori em recém-nascidos.

In vitro antimicrobial effect of bioadhesive oral membrane with chlorhexidine gel

Abstract This study aimed to evaluate in vitro the antimicrobial effect of a bioadhesive chitosan-based oral membrane with chlorhexidine for local treatment of infections in the oral tissues. Five oral membranes of different compositions were tested: 5% chitosan (G1); 5% chitosan ± 0.2% chlorhexidine (G2), 5% chitosan ± 0.6% chlorhexidine (G3), 5% chitosan ± 1.0% chlorhexidine (G4), and 5% chitosan ± 2.0% chlorhexidine (G5). Also, five gel types were tested according to the following compositions: 5% chitosan gel (G6), 0.2% chlorhexidine gel (G7), 2.0% chlorhexidine gel (G8), 5% chitosan gel ± 0.2% chlorhexidine gel (G9), and 5% chitosan gel ± 2.0% chlorhexidine gel (G10). The antimicrobial action of the samples was tested against Candida albicans and Streptococcus mutans through antibiogram by measuring the inhibition halos. Data were statistically analyzed by Kruskal-Wallis and one-way ANOVA followed by Tukey test (p<0.05). The 2.0% chlorhexidine membrane (G5) and the disks containing 2.0% chlorhexidine gel (G8) showed the greatest inhibition halos for both microorganisms, with statistically significant difference when compared to others tested groups (p=0.008) only for Candida albicans inhibitions results. All the other formulations of membranes and gels showed inhibition halos, but without statistically significant difference. The bioadhesive chitosan-based oral membrane with 2% chlorhexidine and 2% chlorhexidine gel were the most effective in inhibiting the tested microorganisms.

Resumo O objetivo deste estudo foi avaliar in vitro o efeito antimicrobiano de uma bandagem oral bioadesiva de quitosana com clorexidina para o tratamento de infecções dos tecidos orais. Cinco bandagens de diferentes composições foram testadas: Quitosana 5% (G1); Quitosana 5% ± clorexidina a 0,2% (G2), Quitosana 5% ± clorexidina a 0,6% (G3), Quitosana 5% ± clorexidina a 1,0% (G4) e Quitosana 5% ± clorexidina a 2,0% (G5). Foram testados também 5 tipos de géis nas seguintes composições: Gel de Quitosana 5% (G6), Gel de clorexidina a 0,2% (G7), Gel de clorexidina a 2,0% (G8), Gel de Quitosana 5% ± clorexidina a 0,2% (G9) e Gel de Quitosana 5% ± clorexidina a 2,0% (G10). A ação antimicrobiana das amostras foi testada contra Candida albicans e Streptococcus mutans por meio do antibiograma, medindo o halo de inibição. Os dados foram analisados pelo teste de Kruskal-Wallis e ANOVA a um critério seguido pelo teste de Tukey (p<0,05). A membrana com 2,0% de clorexidina (G5) e os discos contendo gel com 2,0% de clorexidina (G8) apresentaram os maiores halos de inibição para os dois microrganismos, com diferença estatisticamente significativa em relação aos demais grupos testados (p=0,008) apenas nos resultados de inibição de C. albicans. Todas as outras formulações de membranas e géis apresentaram halo de inibição, mas sem diferença estatisticamente significativa. A bandagem oral bioadesiva de quitosana com gel de 2% de clorexidina foi a mais efetiva em inibir os microrganismos testados.

Factores relacionados a candidiasis oral en niños y adolescentes con VIH, caracterización de especies y susceptibilidad antifúngica / Factors related to oral candidiasis in HIV children and adolescents, species characterization and antifungal susceptibility

Resumen Introducción: Se desconocen los factores asociados a la candidiasis oral en población pediátrica con infección por VIH de los países en desarrollo. Objetivo: Identificar los factores asociados a la colonización por Candida, candidiasis oral y la susceptibilidad in vitro a antifúngicos, en niños y adolescentes con infección por VIH institucionalizados en la ciudad de Tijuana, México. Materiales y Métodos: Se examinó la cavidad oral de 30 niños y adolescentes con infección por VIH, se obtuvo una muestra de la mucosa oral para identificar las especies de Candida mediante cultivo y auxonograma. La susceptibilidad a los antifúngicos se determinó de acuerdo al CLSI. Los indicadores del estado inmunológico y falla virológica se clasificaron conforme a la OMS. Resultados: Se identificaron seis especies de Candida, 53% colonizantes y 47% causantes de candidiasis. Los factores asociados a candidiasis fueron alta carga viral (p = 0,001), menor recuento de LTCD4+ (p = 0,002) y esquema TARAA (p ≤ 0,014). La especie prevalente fue C. glabrata (33%); sin embargo, C. albicans (27%) fue más resistente a fluconazol (p = 0,001). Las especies resistentes a itraconazol se identificaron en esquemas que incluyen un INNTR (p = 0,041). Conclusiones: Los niños y adolescentes con infección por VIH institucionalizados mostraron una prevalencia elevada de Candida spp. colonizante y resistencia a los antifúngicos relacionada con los INNTR .

Background: Factors associated with candidiasis and colonization in HIV-positive children and adolescents in developing countries are not well understood. Aim: To identify the factors associated with oral Candida colonization and candidiasis in institutionalized HIV-positive children and adolescents in Tijuana, México, as well as the response of the isolates to antifungals. Materials and Methods: Sample of the oral mucosa of 30 HIV positive children and adolescents were obtained to isolate and identify Candida species by culture and metabolic profile. Antifungal drugs susceptibility was determined according to CLSI. Indicators of immunological and virologic failure were classified in accordance to WHO criteria. Results: Six Candida species were identified from oral mucosa, 53% colonizers and 47% in candidiasis. Factors associated with candidiasis and oral colonization were viral load (p = 0,001), CD4+ counts (p = 0,002) and HAART regimen (p ≤ 0,014). The most prevalent species was C. glabrata (33%), but C. albicans (27%) was more resistant to fluconazole (p = 0,001). Itraconazol resistant species were identified in regimens that include an NNRTI (p = 0,041). Conclusion: HIV-positive children and adolescents living in an orphanage showed high prevalence of colonizing Candida spp. and resistance to antifungals, related to NNRTI.

Phenotypic and genotypic detection of Candida albicans and Candida dubliniensis strains isolated from oral mucosa of AIDS pediatric patients

ABSTRACT The aim of this study was to assess a collection of yeasts to verify the presence of Candida dubliniensis among strains isolated from the oral mucosa of AIDS pediatric patients which were initially characterized as Candida albicans by the traditional phenotypic method, as well as to evaluate the main phenotypic methods used in the discrimination between the two species and confirm the identification through genotypic techniques, i.e., DNA sequencing. Twenty-nine samples of C. albicans isolated from this population and kept in a fungi collection were evaluated and re-characterized. In order to differentiate the two species, phenotypic tests (Thermotolerance tests, Chromogenic medium, Staib agar, Tobacco agar, Hypertonic medium) were performed and genotypic techniques using DNA sequencing were employed for confirmation of isolated species. Susceptibility and specificity were calculated for each test. No phenotypic test alone was sufficient to provide definitive identification of C. dubliniensis or C. albicans, as opposed to results of molecular tests. After amplification and sequencing of specific regions of the 29 studied strains, 93.1% of the isolates were identified as C. albicans and 6.9% as C. dubliniensis. The Staib agar assay showed a higher susceptibility (96.3%) in comparison with other phenotypic techniques. Therefore, genotypic methods are indispensable for the conclusive identification and differentiation between these species.

Candida spp. adherence to oral epithelial cells and levels of IgA in children with orthodontic appliances

Adhesion and colonization of the oral cavity by Candida albicans is an initial step in candidosis. Orthodontic and other oral appliances seem to favor candidal presence. The aim of this work was to compare the presence of Candida species in saliva, their adherence to oral epithelial cells, and the levels of anti-C. albicans IgA in children with or without orthodontic appliances. This study included 30 children 5 to 12 years old (9.1 ± 1.7 years old) who were users of removable orthodontic devices for at least 6 months and 30 control children of similar ages (7.7 ± 1.5 years old). The presence of yeast species in the saliva was evaluated by microbiological methods. Candida species were identified using phenotypic methods. Anti-C. albicans IgA levels in saliva were analyzed by ELISA. The yeasts adhering to oral epithelial cells were assessed by exfoliative cytology. No statistically significant differences were observed for saliva yeast counts and anti-C. albicans IgA levels between the studied groups. Children with orthodontic devices exhibited more yeast cells adhering to oral epithelial cells and a higher percentage of non-albicans species relative to the control group. In conclusion, orthodontic appliances may favor the adherence of Candida to epithelial cells but do not influence the presence of these yeasts in saliva, and the levels of anti-C. albicans IgA do not correlate with yeast adherence or presence of Candida in the oral cavity.

Resposta imune inata na estomatite protética: interação in vitro entre células epiteliais de palato humano e Candida albicans / Innate imune response in denture stomatitis: in vitro interaction between human palatal epithelial cells and Candida albicans

A presença de Candida albicans nos biofilmes microbianos da superfície interna das próteses totais superiores está relacionada com uma doença inflamatória no palato, a estomatite protética. Constituinte da defesa inata do hospedeiro, o epitélio bucal, por sua vez, tem a capacidade de reconhecer e reagir aos fatores fúngicos a fim de evitar a invasão pelo microrganismo. O objetivo deste trabalho foi avaliar in vitro o efeito direto e indireto de C. albicans viável sobre as células epiteliais de palato humano (CEPH) ao longo do tempo. Objetivamos correlacionar os eventos de agressão, apoptose e invasão das CEPH provocados pelo fungo, com as respostas de defesa epitelial mediante produção de óxido nítrico (NO) e expressão gênica do peptídeo antimicrobiano β-defensina 2 (hBD-2). Material e Métodos: As CEPH foram obtidas, parte pelo método explante e parte pelo método enzimático, e mantidas em co-cultivo sobre uma camada de sustentação feederlayer (fibroblastos gengivais humanos mitoticamente inativados). Após desafios das CEPH com C. albicans ATCC 90028 por contato direto fungo-epitélio (D.D.) e indireto pelo sobrenadante da cultura do fungo hifal (D.I.), proporções de desafio de 0,01/1; 0,025/1 e 0,1/1 levedura/queratinócito (FUN/EPI) e tempos experimentais de 3, 6 e 10 h foram determinados; via ensaios de viabilidade celular por imunofluorescência (LIVE/DEAD), e análise qualitativa da invasão celular pelo fungo por meio do método colorimétrico com laranja de acridina. A apoptose epitelial foi determinada pela marcação nuclear fluorescente com Hoechtst 33258. A produção de óxido nítrico (NO) e a expressão de RNAm de hBD-2 foram avaliados por reação colorimétrica de Griess e RT-qPCR, respectivamente. Os resultados foram expressos como média ± desvio padrão e submetidos aos testes estatísticos ANOVA Fatorial, Teste de Contraste; ou Teste de Mann-Whitney (p<0,05). Resultados: Em 3 h, foi detectado aumento da apoptose das células epiteliais em relação ao...

The presence of the fungus Candida albicans in the microbial biofilm underlying maxillary prosthesis is related to an inflammatory reaction of the palatal mucosa, the denture stomatitis. As a component of the host innate defense, the oral epithelium has the ability to recognize and react to fungal factors in order to prevent the microrganism invasion. The aim of this study was to in vitro evaluate the direct and indirect effect of viable C. albicans on the human palatal epithelial cells (HPEC) over time. The aggressive events, such as apoptosis and HPEC invasion by the fungus, were correlated with epithelial defense responses through the nitric oxide (NO) production and antimicrobial peptides β-defensin (hBD-2) mRNA expression. Methods: The HPEC were obtained by explant and enzymatic methods, and were maintained in co-culture on a feeder-layer support (mitotically inactivated human gingival fibroblasts). After the HPEC challenges with C. albicans ATCC 90028 by direct contact fungus-epithelium (D.D.) and indirect contact by supernatant from hyphal fungus (D.I.), defiance ratios of 0.01/1, 0.025/1 and 0.1/1 yeast/keratinocyte (FUN/EPI) and experimental times of 3, 6 and 10 h were determined. These conditions were standardized by cell viability immunofluorescence assay (LIVE/DEAD), and cell invasion qualitative analysis (colorimetric method with acridine orange). The apoptotic cells were determined by fluorescent nuclear staining with Hoechtst 33258. The nitric oxide (NO) production and hBD-2 gene expression were evaluated by Griess colorimetric reaction and RT-qPCR, respectively. The results were expressed as mean ± standard deviation and were analyzed using the factorial ANOVA, Contrast Test; or Mann-Whitney Test (p<0,05). Results: At 3 h, the apoptotic epithelial cells under 0.1/1 FUN/EPI increased compared to epithelium unchallenged (p<0,05) that remained over time with increasing concentration and independent of D.D. and D.I. The onset...

Fungal infections of the oral mucosa.

Fungal infections in humans occur as a result of defects in the immune system. An increasing emergence in oral Candidal and non-Candidal fungal infections is evident in the past decade owing to the rise in the immunodeficient and immunocompromised population globally. Oral Candidal infection usually involves a compromised host and the compromise may be local or systemic. Local compromising factors include decreased salivation, poor oral hygiene, wearing dentures among others while systemic factors include diabetes mellitus, nutritional deficiency, HIV infection/AIDS and others. Oral candidiasis is generally a localized infection and rarely appears as a systemic fungal disease whereas oral non-Candidal fungal infections are usually signs of disseminated disease. Some of the non-Candidal fungi that were once considered exotic and geographically restricted are now seen worldwide, beyond their natural habitat, probably attributed to globalization and travels. Currently infections from these fungi are more prevalent than before and they may present either as primary oral lesions or as oral manifestations of systemic mycoses. This review discusses the various predisposing factors, clinical presentations, clinical differential diagnosis, diagnosis and management of oral candidiasis, as well as briefly highlights upon a few of the more exotic non-Candidal fungi that infect the oral mucosa.

Candida bucal en niños mexicanos con VIH/sida, desnutrición o marginación social / Oral candida in Mexican children with malnutrition, social marginalization, or HIV/AIDS

OBJETIVO: Determinar la frecuencia de Candida en cavidad bucal de niños con riesgo de desarrollar infecciones oportunistas y establecer si existe asociación entre la frecuencia de esta colonización bucal y tres tipos de población en riesgo. MÉTODOS: Se estudiaron cuatro grupos de población infantil de México: grupo VIH/sida bajo terapia antirretroviral altamente activa (TAAA) (35 niñas y 25 niños); grupo desnutrición (26 niñas y 29 niños); grupo tarahumara (37 niñas y 20 niños), una de las poblaciones étnicas más pobres del país, y grupo control (8 niñas y 21 niños aparentemente sanos). Los niños con VIH/sida fueron inmunológica y virológicamente clasificados según los criterios de EC-Clearinghouse, mientras que la desnutrición fue determinada a través del índice peso/talla de la Organización Mundial de la Salud. Se tomó una muestra de la mucosa bucal con hisopo estéril, que fue incubada en agar dextrosa Sabouraud y en CHROMagar-Candida®. Las especies de Candida se confirmaron con la prueba API ID32C. RESULTADOS: Los grupos VIH/sida y desnutrición mostraron la frecuencia más alta de Candida spp. (51,7 por ciento y 38,2 por ciento, respectivamente) mientras que el grupo tarahumara presenta una frecuencia semejante a la del grupo control (17,5 por ciento vs 10,3 por ciento). Respecto a las especies de Candida, el grupo desnutrición mostró la mayor diversidad: C. albicans, C. tropicalis, C. krusei y C. glabrata. CONCLUSIONES: Los infantes con inmunodeficiencia y con desnutrición requieren de estrategias diseñadas para disminuir la colonización bucal candidal y disminuir el riesgo de infecciones oportunistas.

OBJECTIVE: Determine the frequency of candida in the oral cavity of children with a risk of developing opportunistic infections, and establish if there is an association between the frequency of this oral colonization and three categories of at-risk populations. METHODS: Four infant population groups in Mexico were studied: an HIV/AIDS group undergoing highly active antiretroviral therapy (35 girls and 25 boys); a malnourished group (26 girls and 29 boys); a group from the Tarahumara indigenous people, one of the poorest ethnic populations in the country (37 girls and 20 boys); and a control group (8 girls and 21 boys in apparently good health). The children with HIV/AIDS were immunologically and virologically classified according to the EC Clearinghouse criteria, while malnutrition was determined through the World Health Organization's weight/height index. A sample of oral mucosa was taken with a sterile swab, which was incubated in Sabouraud dextrose agar and in Candida CHROMagar®. The species of candida were confirmed through the API ID32C test. RESULTS: The HIV/AIDS and malnutrition groups showed the higher frequency of Candida spps (51.7 percent and 38.2 percent, respectively), while the frequency level in the Tarahumara group was similar to that of the control group (17.5 percent versus 10.3 percent). With regard to the species of candida, the malnutrition group had the greatest diversity: C. albicans, C. tropical, C. krusei, and C. glabrata. CONCLUSIONS: The children with HIV/AIDS and malnutrition require strategies designed to reduce oral candidal colonization and reduce the risk of opportunistic infections.

A retrospective study of oral manifestations in patients with paracoccidioidomycosis

South American blastomycosis, paracoccidioidomycosis (Pb mycosis) or Lutz disease is an endemically fungal infection in Latin America. It is caused by the dimorphic fungus Paracoccidioides brasiliensis and may cause oral mucosal lesions. The incidence of Pb mycosis oral lesions was evaluated in patients assisted at a Brazilian Dental School's Specialized Oral Diagnosis Service with special focus on the different clinical forms of these lesions, its location, patients' occupation, deleterious habits, and diagnosis methodology. Students' and professionals' initial diagnoses were compared with the definitive diagnosis. Lesions were detected 31 cases (18 patients). The results show that 88.8% of the patients were male with a mean age of 50 years and 39% work(ed) with activities related to agriculture. As much as 88.9% were smokers and 72.2% were alcohol users. Exfoliative cytology was performed in 66.6% of the patients. Oral mucosa (30%), gingiva (16.6%) and lips 16.6% were the most common sites of Pb mycosis oral lesions. Comparing the initial with the definitive diagnosis made by the professionals their accuracy was 33% (6 out of 18 patients). Students' diagnosis was more accurate demonstrating 72.5% of initial correct diagnosis (13 out of 18). Statistical analysis by ANOVA (α=0.05, SPSS WIN) demonstrated a significant difference between the diagnosis of Pb mycosis made by students and professionals when considering initial diagnosis and final diagnosis (after histopathological analysis) (p=0.25). Incisional biopsy and exfoliate cytology are efficient for an early diagnosis of this disease in mouth. Students' training in diagnosis of oral pathologies to recognize lesions is urgent to improve public health.

A blastomicose sul americana, paracoccidiodomicose (Pb micose) ou doença de Lutz é uma infecção fúngica endêmica na América Latina. É causada pelo fungo dimórfico Paracoccidiodes brasiliensis, e pode causar lesões na mucosa oral. Nós avaliamos a incidência de lesões orais de Pb micose nos pacientes atendidos em um Centro Especializado em Diagnóstico Oral com foco principal nas diferentes formas clinicas das lesões, suas localidades, ocupação dos pacientes, hábitos deletérios e diagnóstico e metodologia, e foram detectados 31 casos (18 pacientes). Os resultados mostraram que 88,8% dos pacientes eram do sexo masculino com idade média de 50 anos de idade, e 39% trabalham, ou trabalhavam, com atividades relacionadas à agricultura. Observou-se que 88,9% eram fumantes e 72,2% ingeriam álcool. Citologia esfoliativa foi feita em 66,6%. Mucosa jugal foi acometida em 30%, gengiva, e lábios 16% (cada um) foram os locais mais comuns de lesões orais da Pb micose. Comparando o diagnóstico inicial com o definitivo feito pelos profissionais, a acurácia foi de 33%; o diagnóstico dos estudantes foi mais preciso demonstrando 72,5% do diagnóstico inicial correto com diferença estatística significante (p=0,25) através do teste ANOVA do SPSS WIN com nível de significância de 5%. Biópsia incisonal e citologia esfoliativa são eficientes para um diagnóstico precoce desta doença na boca; o treinamento dos estudantes em diagnóstico para reconhecer as patologias orais é urgente para melhorar a saúde pública.

In vitro adherence of Candida albicans isolated from patients with chronic periodontitis

Adherence is considered an extremely important virulence factor in yeast. OBJECTIVE: The aim of this study was to analyze the adherence to epithelial cells of C. albicans isolated from patients with chronic periodontitis in comparison to healthy patients. MATERIAL AND METHODS: Candida albicans cells isolated from individuals with chronic periodontitis (n=25) and healthy controls (n=25) were included in this study. Suspensions of C. albicans (10(6) cells/mL) and epithelial cells (10(5) cells/mL) were mixed and incubated at 37ºC for 1 h. The number of yeasts adhered to 25 epithelial cells was counted. RESULTS: The number of C. albicans cells adhered to epithelial cells was statistically higher in the chronic periodontitis group than in the control group (Student's t-test, p=0.000). CONCLUSION:The results of the present study suggest a higher Candida adherence of samples isolated from patients with chronic periodontitis.

Efectividad antimicrobiana del colutorio de matricaria recutita, en funcionarios de la Facultad de Odontología de la Universidad del Desarrollo, Chile / Antimicrobial Effectiveness of Matricaria recutita Mouthwash in the Dentistry Faculty Employees of Universidad del Desarrollo, Chile

El colutorio de manzanilla es un producto natural, no produce tinciones, alteraciones del sabor, no es toxico y puede ser utilizado por pacientes embarazadas, adultos mayores y niños. Se realizó un estudio experimental, con el objetivo de determinar la efectividad antimicrobial del colutorio en base a el extracto de Matricaria recutita l, tipo manzanilla primavera Puelche, el cual fue fabricado por la Dirección de Ciencias Básicas de la Facultad de Odontología de la Universidad del Desarrollo, Chile. Mediante un muestreo no probabilístico se reclutó 32 pacientes, funcionarios de la Facultad de Odontología. La metodología consignó la aplicación del Colutorio de Manzanilla (CM) en 5 pacientes (grupo experimental), Suero Fisiológico (SF) en 6 pacientes (control negativo) y Clorhexidina al 0,12 por ciento (CX) en 7 pacientes (control positivo). Se tomó muestras de la mucosa del carrillo y de superficie vestibular del primer molar superior, previo a la aplicación del colutorio y en 7 intervalos de tiempo; se cultivó en condiciones de aerobiosis a 37 °C por 48 horas. Luego se realizó el recuento bacteriano. Los resultados se analizaron mediante ANOVA para varianzas homogéneas y Kruskall-Wallis para varianzas heterogéneas. Se demostró que el recuento bacteriano no presenta diferencias significativas en los tiempos analizados para mucosa (CM pvalue: 0,2507. CX pvalue: 0,1769. SF pvalue: 0,9397) y para diente (CM pvalue: 0,2540. CX pvalue: 0,2859. SF pvalue: 0,3471), observándose que ningún resultado otorga un pvalue < a 0,05, en los 7 tiempos de aplicación. De esta manera se concluyó que la frecuencia de uso clínico del CM, presenta una mayor disminución de carga bacteriana cada 4 a 6 horas.

Chamomilemouthwashis a natural product,does not causestains, taste disturbances,is non-toxic andcan be used bypregnant patients, the elderlyand children. An experimental studywas performedwith the aimof determining theantimicrobial effectivenessofmouthwashproduced fromMatricariarecutitaextractl, Puelche Manzanilla Primavera Type, which wasproducedby the Department ofBasic Sciences, of the Dentistry Faculty of the Universidad del Desarrollo.Thirty two patients were recruited using anon-probabilitysampling, employees from theSchool of Dentistry.The methodologyof work was defined by the application of chamomilemouthwash(CM) in 5 patients(experimental group),saline(SF) in 6patients (negative control) and0.12 percent chlorhexidine(CX) in 7patients (positive control).Samples were taken from the cheek mucosa and upper first molar bucal surface, prior to the application of chamomille mouthwash and 7 time intervals. The samples were grown under aerobic conditions at 37°C for 48 hours. This was followed by bacterial count. The results, like homogeneous variance, were analyzed by ANOVA and Kruskall-Wallis to heterogeneous variances.It was shown thatthe bacterial countdid not differ significantly with the timeanalyzed formucosa (CMpvalue:0.2507. CX pvalue: 0.1769, Pvalue SF: 0.9387 and tooth (CM pvalue <0.2540). CX pvalue: 0.2859. pvalue SF: 0.3471) showing that no resultgives apvalue<0.05,within 7days ofapplication. Thusit was concludedthat the frequency ofclinical use ofchamomilemouthwash, presentsa greater reduction inbacterial loadevery 4 to6 hours.

Antibiotic susceptibility of Staphylococcus aureus isolates in oral mucosa and pockets of patients with gingivitis-periodontitis

La cavidad bucal y el interior de la bolsa subgingival constituyen nichos ecológicos propicios para albergar microorganismos que podrían actuar como patógenos oportunistas, como el Staphylococcus aureus y enparticular S. aureus resistente a la meticilina (SARM). La detección temprana de portadores reviste importancia para la salud pública. El objetivo de nuestro trabajo fue determinar por métodos fenotípicos y genotípicos la meticilino resistencia de cepas de S. aureus aisladas de mucosa bucal y bolsa subgingival y bolsa subgingival de 102 pacientes con enfermedad gingivoperiodontal. Se observó una prevalencia de S. aureus en bolsa subgingival del 10,8 por ciento (n=11) y en mucosa bucal del 19,5 por ciento (n=20). Se obtuvieron 31 aislamientos de S. aureus de los cuales 13 fueorn mec A positivos y 18 eran mec A negativos. La detección del gen mec A por PCR se utilizó como método de referencia para comparar los resultados de métodos fenotípicos para determinar la resistencia a meticilina. La detección rápida y exacta de S. aureus por métodos microbiológicos fenotípicos y genotípicos es relevante para evaluar la colonización y prevenir la propagación del SARM.

Detection of selected bacterial species in intraoral sites of patients with chronic periodontitis using multiplex polymerase chain reaction

OBJECTIVE: The aim of this study was to detect the prevalence of selected bacterial species in intraoral sites of patients with chronic periodontitis (CP) using multiplex polymerase chain reaction (PCR). METHODOLOGY: Samples were collected from the tongue dorsum, buccal mucosa, supragingival and subgingival plaque and saliva of 30 patients with untreated CP. Multiplex PCR was used to determine prevalence rates, which were then compared using a chi-square test. Significance level was set at p<0.05. Mean and standard deviation values were used to evaluate variations in prevalence according to site. RESULTS: The prevalence of S. mutans was 70 percent in saliva; 60 percent in samples collected from the tongue dorsum; 50 percent in samples collected from the buccal mucosa; 56.5 percent in the supragingival plaque; and 53.5 percent in the subgingival plaque. The prevalence of E. faecalis ranged from 3.5 percent to 13.5 percent in all intraoral microenvironment. The highest prevalence of P. gingivalis was found in subgingival plaque (53.5 percent), and of P. intermedia in supragingival plaque (33.5 percent), subgingival plaque (30 percent) and tongue dorsum (33.5 percent). The prevalence of bacteria did not vary significantly among the intraoral sites. CONCLUSIONS: All studied bacteria were identified in intraoral sites. S. mutans, P. gingivalis and P. intermedia had high prevalence rates, but the prevalence of E. faecalis was low. Multiplex PCR proved to be an adequate method for epidemiological studies.

Enhanced susceptibility of Candida albicans to chlorhexidine under anoxia

Aim: Periodontal pockets can be colonized not only by bacteria, but also by Candida albicans. However, its role in periodontitis is unknown. This study evaluated the inhibitory performance of chlorhexidine digluconate under normoxic and anoxic conditions against 16 strains of C. albicans from periodontal pockets and other 20 from the oral mucosa. Methods: Strains were grown in normoxia and anoxia to adapt themselves to the different atmospheric conditions. Microdilution-based assays were carried out to determine the minimum concentrations of chlorhexidine that may restrain the conditioned candidal strains, in normoxia (normoxic MIC) and anoxia (anoxic MIC). The Mann-Whitney U test was used to evaluate the antimicrobial effect of chlorhexidine on C. albicans under normoxic and anoxic conditions (α = 0.05). Results: The normoxic MIC of chlorhexidine varied broadly from 150 to 1200 µg/mL, whereas its anoxic MIC varied narrower from 2.34 to 37.5 µg/mL. Regarding the origins of strains, no statistically significant differences (p > 0.05) were found. Conclusions: These results indicate that anoxic environmental conditions, compatible with periodontal pockets, tend to enhance C. albicans susceptibility to chlorhexidine.

Biotypes of Candida albicans isolates from the oral mucosa of HIV seropositive and control subjects / Biótipos de amostras de Candida albicans isoladas da mucosa oral de sujeitos HIV soropositivos e controles

Purpose: This study evaluated the Candida albicans biotypes from oral mucosa according to some host variables, such as HIV infection; medication use - protease inhibitors (PI), non protease inhibitors (NPI) or no medication (NM); dental prosthesis wearing (PW) or not (NPW); and yeast variables (activity levels of protease and phospholipase). Methods: Samples from the oral mucosa of 193 HIV+ subjects and 205 HIV- subjects were collected by means of sterile swabs and seeded onto Sabouraud dextrose agar. The isolates were identified by microculture on slide, germ tube formation, auxanogram, and zimogram. Ninety-two isolates were obtained from HIV+ individuals: 49 from patients under PI, 31 from patients under NPI and 12 from patients with no medication. The control group comprised 63 isolates from HIV- patients. Results: From the 95 possible C. albicans biotypes, 46 were identified in the sample, and the most prevalent were: 10122 (46.4%); 11122 (38.01%); 01031 (42.4%); 00022 (40%); (P<0.01 - Spearman correlation test). No difference was detected among PI, NPI, and NM groups. The control group exhibited intermediate enzymatic activity level from dentate isolates, and high protease activity level amongst isolates from prosthesis wearers.Conclusion: It was not possible to detect any inhibitory action of PI drugs on the enzymatic activity of C. albicans.

Objetivo: Este estudo avaliou biótipos de isolados de Candida albicans da mucosa oral utilizando-se variáveis do hospedeiro: infecção pelo HIV; medicação - uso de inibidores de protease (IP), não inibidores (NIP), ausência de medicação (SM); e uso ou não de prótese. Da levedura foram utilizadas as variáveis relativas à produção de protease e fosfolipase (nível de atividade). Metodologia: O material da mucosa bucal de 193 sujeitos HIV+ e 205 sujeitos HIV- foi coletado com swab estéril e semeado em Agar Sabouraud dextrose. A identificação dos isolados foi obtida pelos testes de micro cultivo em lâmina, tubos germinativos, auxanograma e zimograma. Utilizaram-se 92 amostras isoladas de indivíduos HIV+, sendo 49 de pacientes IP, 31 de pacientes NIP e 12 de pacientes sem uso de medicação (SM). O controle constou de 63 amostras isoladas de indivíduos HIV-. Resultados: Dos 95 diferentes biótipos possíveis, obteve-se 46, sendo os mais prevalentes: 10122 (46,4%); 11122 (38,01%); 01031 (42,4%); 00022 (40%); (P<0,01 - teste de correlação de Spearman). Não houve nenhuma diferença entre os subgrupos IP, NIP e SM. As amostras isoladas dos controles exibiram atividade intermediária de protease (dentados) e alta atividade de protease (portadores de próteses). Conclusão: Não foi possível constatar interferência dos IP na atividade enzimática de C. albicans.

Cell surface hydrophobicity and adherence of a strain of group B streptococci during the post-antibiotic effect of penicillin / Hidrofobicidade de superfície celular e aderência de uma estirpe de estreptococos do grupo B durante o efeito pós-antibiótico da penicilina

The minimum inhibitory concentration and post-antibiotic effects of an antimicrobial agent are parameters to be taken into consideration when determining its dosage schedules. The in vitro post-antibiotic effects on cell surface hydrophobicity and bacterial adherence were examined in one strain of group B streptococci. Exposure of the microorganism for 2 h at 37 °C to 1 x MIC of penicillin induced a PAE of 1.1 h. The cell surface charge of the Streptococcus was altered significantly during the post-antibiotic phase as shown by its ability to bind to xylene: hydrophobicity was decreased. Bacterial adherence to human buccal epithelial cells was also reduced. The results of the present investigation indicate that studies designed to determine therapeutic regimens should evaluate the clinical significance of aspects of bacterial physiology during the post-antibiotic period.

A concentração mínima inibitória e os efeitos pós-antibióticos (EPA) de um agente antimicrobiano são parâmetros que devem ser levados em consideração quando da determinação do esquema de dosagem. Os efeitos pós-antibióticos in vitro na hidrofobicidade de superfície celular e na aderência foram pesquisados em uma amostra de estreptococos do grupo B. A exposição do microrganismo por 2 h a 37 °C a 1 x CMI de penicilina induziu um EPA de 1,1 h. A carga da superfície celular da bactéria foi alterada significativamente durante a fase pós-antibiótica revelada através da capacidade de ligação ao xileno, indicada pela diminuição da hidrofobicidade. A aderência bacteriana às células epiteliais bucais humanas também foi reduzida. Os resultados da investigação demonstram que estudos clínicos destinados a determinar regimes terapêuticos deveriam incluir o conhecimento da fisiologia bacteriana durante o período pós-antibiótico.

Etiology and outcome of oral mucosal lesions in children on chemotherapy for acute lymphoblastic leukemia.

Microbiological cultures were taken from oral cavity and blood in 100 mucositis episodes in 70 children with acute lymphoblastic leukemia (ALL). Oral mucositis was commonest in neutropenic children during induction chemotherapy. Fungal organisms (n=39) were commonest isolate from mucosa followed by bacteria (n=28). Isolation of organism from oral cavity had no association with those isolated from blood. Herpes serology was positive in 16% episodes compared to 2% of controls. Obtaining cultures from oral lesions is useful in appropriate management of lesions and thereby possibly preventing systemic spread.

Oral candidiasis in HIV+ patients under treatment with protease inhibitors

The purpose of this work was to evaluate the influence of Protease Inhibitors (PI) on the occurrence of oral candidiasis in 111 HIV+ patients under PI therapy (Group A). The controls consisted of 56 patients that were not using PI drugs (Group B) and 26 patients that were not using any drugs for HIV therapy (Group C). The patient's cd4 cell counts were taken in account for the correlations. One hundred and ninety three patients were evaluated. The PI did not affect the prevalence of oral candidiasis (p = 0.158) or the frequency of C. albicans isolates (p = 0.133). Patients with lower cd4 cell counts showed a higher frequency of C. albicans isolates (p = 0.046) and a greater occurrence of oral candidiasis (p = 0.036).