Contaminação microbiana em colírios de pacientes em tratamento de glaucoma / Microbial contamination in eye drops of patients in glaucoma treatment

Resumo Objetivos: Avaliar o grau de contaminação por fungos e bactérias e o modo de conservação destes colírios hipotensores por parte dos pacientes no ambulatório de Glaucoma da Santa Casa de Ribeirão Preto. Métodos: Foram selecionados aleatoriamente cinquenta e cinco pacientes, em seguimento no ambulatório, e após consentimento dos mesmos os colírios eram coletados e enviados via correio para análise por microbiologista e patologista em até 72 horas. Foi analisado 0,5ml aproximadamente das medicações e os pacientes respondiam a um questionário simples sobre o método de conservação e se consideravam estes adequados. Resultados: Dos 55 colírios analisados, cinco (9,01%) estavam com seu conteúdo líquido contaminado. Entre os microrganismos isolados haviam 4 bactérias Gram negativas, sendo 1 (1,8%) por Serratia marcescens, 1 (1,8%) Pseudomonas aeruginosa e 2 (3,6%) Stenotrophomas maltophilia. Um colírio estava contaminado pelo fungo Cândida ssp Todos pacientes do estudo julgam seus métodos de armazenamento e instilação adequados. Os pacientes que tiveram os colírios positivados eram convocados para exame clínico e passavam por novo questionário pelo investigador. Conclusão: O tempo de abertura dos frascos e os métodos de conservação influenciam na contaminação dos medicamentos, todos os colírios com crescimento de microrganismos no presente estudo estavam abertos entre 30 e 90 dias. O fato de que a maioria dos pacientes levam seus colírios em tarefas cotidianas, aumenta a exposição dos frascos e podem ser um fator relevante para determinar a contaminação destas medicações.

Abstract Objetives: To assess the degree of fungal and bacterial contamination of hypotensive eye drops and the way these are preserved by the patients at the Glaucoma outpatient clinic of Santa Casa Hospital in Ribeirão Preto. Methods: Fifty-five patients were randomly assigned to follow-up in the outpatient clinic and, after their consent, an eye drop was collected per patient and later sent by mail for analysis by microbiologist and pathologist in up to 72 hours. Approximately 0.5ml of the medications were analyzed and the patients were asked to answer a simple questionnaire on the method of drug conservation and whether they considered it adequate. Results: Of the 55 analysed eye drops, five (9.01%) had their liquid contents contaminated. Among the microorganisms isolated there were 4 Gram negative bacteria, 1 (1.8%) by Serratia marcenses, 1 (1.8%) Pseudomonas aeruginosa and 2 (3.6%) Stenotrophomas maltophilia. An eye drop was contaminated by the fungus Candida ssp. All the patients in the study judged their methods of storage and instillation appropriate. The patients who had the positive coliria were summoned for clinical examination and passed through a new questionnaire by the investigator. Conclusion: The time and methods of preservation influence the contamination of medicinal products. All the eye drops that presented growth of microorganisms in the present study were open between 30 and 90 days. The fact that most patients take their eye drops on daily tasks increases the exposure of the bottles and can be a relevant fact to determine the contamination of these medications.

Evaluation of the frequency of Candida spp. in hospitalized and non-hospitalized subjects / Avaliação da frequência de candida spp. em indivíduos hospitalizados e não hospitalizados

Abstract The aim of this study was to evaluate the frequency of Candida species between a non-hospitalized and a hospitalized population. For this purpose, samples of saliva were sampled through sterile swabs, moistened in peptone water and rubbed in the oral cavity of 140 individuals, from which, 70 were hospitalized patients from the Medical Clinic of a Teaching Hospital and the other 70 were non-hospitalized subjects. All saliva samples were plated in Sabouraud Dextrose agar added with Chloramphenicol and incubated at 36 °C for 48 hours. The morphology identification was performed through macroscopic and microscopic characterization, the CHROMagar Candida medium and the VITEK® system Yeast Biochemical Card (bio Mérieux SA, France). The results showed a colonization of Candida spp. in 85.7% the hospitalized individuals, where the species found were C. albicans (60%), C. tropicalis (23.4%), C. krusei (3.3%) and Candida spp. (13.3%). In the non-hospitalized individuals the colonization by Candida spp was 47.1%, and the species found were: C. albicans (45.5%), C.krusei (9.1%), C. guilliermondii (9.1% %), C. tropicalis (3.0%), C. famata (3.0%) and Candida spp. (30.3%). In spite of their presence in oral cavity in both groups, Candida spp. was more frequently isolated in hospitalized individuals, who were 6.73 times more likely to have this fungus in the oral cavity and were 3.88 times more likely to have Candida albicans.

Resumo O objetivo deste estudo foi avaliar a frequência de espécies de Candida entre uma população de indivíduos não-hospitalizados e hospitalizados. Para isto, amostras de saliva foram coletadas através de swabs estéreis, umedecidas em água de peptona e friccionadas na cavidade bucal de 140 indivíduos, dos quais 70 eram pacientes internados em uma Clínica Médica de um Hospital Escola e os outros 70 eram indivíduos não hospitalizados sem contato com ambiente hospitalar. Todas as amostras de saliva foram plaqueadas em ágar Sabouraud dextrose adicionadas de cloranfenicol e incubadas a 36 °C durante 48 horas. A identificação morfológica foi realizada através da caracterização macroscópica e microscópica, com o meio CHROMagar Candida e do sistema VITEK® Biochemical Card (bio Mérieux SA, França). Os resultados mostraram uma colonização de Candida spp. em 85,7% dos indivíduos hospitalizados, onde as espécies encontradas foram: C.albicans (60%), C. tropicalis (23,4%), C. krusei (3,3%) e Candida spp. (13,3%). Nos indivíduos não-hospitalizados a colonização por Candida spp foi de 47,1%, e as espécies encontradas foram: C. albicans (45,5%), C. krusei (9,1%), C. guilliermondii (9,1%), C. tropicalis (3,0%), C. famata (3,0%) e Candida spp. (30,3%). Apesar de sua presença na cavidade oral em ambos os grupos, Candida spp. foi mais freqüentemente isolada em indivíduos hospitalizados, que foram 6,73 vezes mais propensos a ter este fungo na cavidade oral e foram 3,88 vezes mais propensos a ter Candida albicans.

Role of Candida species from HIV infected children in enamel caries lesions: an in vitro study

Abstract Objectives This study analyzed the capacity of Candida spp. from dental biofilm of HIV infected (HIV+) children to demineralize primary molar enamel in vitro by Transversal Microhardness (TMH), Polarized Light Microscopy (PLM) and the quantity of calcium ions (Ca2+) released from the enamel. Material and Methods Candida spp. samples were isolated from the supragingival biofilm of HIV+ children. A hundred and forty (140) enamel blocks were randomly assigned to six groups: biofilm formed by C. albicans (Group 1); mixed biofilm formed by C. albicans and C. tropicalis (Group 2); mixed biofilm formed by C. albicans and C. parapsilosis (Group 3); mixed biofilm formed by C. albicans, C. parapsilosis and C. glabrata (Group 4); biofilm formed by C. albicans ATCC (Group 5) and medium without Candida (Group 6). Enamel blocks from each group were removed on days 3, 5, 8 and 15 after biofilm formation to evaluate the TMH and images of enamel were analyzed by PLM. The quantity of Ca2+ released, from Groups 1 and 6, was determined using an Atomic Absorption Spectrophotometer. The SPSS program was used for statistical analysis and the significance level was 5%. Results TMH showed a gradual reduction in enamel hardness (p<0.05) from the 1st to 15th day, but mainly five days after biofilm formation in all groups. The PLM showed superficial lesions indicating an increase in porosity. C. albicans caused the release of Ca2+ into suspension during biofilm formation. Conclusion Candida species from dental biofilm of HIV+ children can cause demineralization of primary enamel in vitro.

Quercetin and rutin as potential agents antifungal against Cryptococcus spp / Quercetina e rutina: potenciais agentes para terapia antifúngica

Abstract Amphotericin B is a fungicidal substance that is treatment of choice for most systemic fungal infections affecting as cryptococcosis the immunocompromised patients. However, severe side effects have limited the utility of this drug. The aim of this study was to evaluate the antifungal effect of the combination of amphotericin B with quercetin or rutin and as a protective of citotoxic effect. The antifungal activity to amphotericin B, quercetin and rutin alone and in combination was determined in Candida sp and Cryptococcus neoformans strains. Cytotoxicity test on erythrocytes was performed by spectrophotometric absorbance of hemoglobin. The amphotericin B MIC was reduced when used in combination with quercetin or rutin to C. neoformans ATCC strain and reduced when combined with rutin to a clinical isolate of C. neoformans. In addition, the combination of quercetin with amphotericin B may reduce the toxicity of amphotericin B to red blood cells. Our results suggest that quercetin and rutin are potential agents to combine with amphotericin B in order to reduce the amphotericin dose to lessen side effects and improve antifungal efficacy.

Resumo A anfotericina B é uma substância fungicida e é o tratamento de escolha para a maioria das infecções fúngicas sistêmicas que afetam os pacientes imunocomprometidos, como a criptococose. No entanto, as severas reações adversas têm limitado a utilização desta droga. O objetivo deste estudo foi avaliar o efeito antifúngico e o potencial efeito protetor de citotoxicidade da combinação de anfotericina B com quercetina ou rutina. A atividade antifúngica de anfotericina B, quercetina e rutina, isoladamente e em combinação foi determinada em cepas de Candida sp e Cryptococcus neoformans. O teste de citotoxicidade em eritrócitos foi realizado por espectrofotometria, através da determinação da absorbância da hemoglobina. A concentração inibitória mínima da anfotericina B foi reduzida quando utilizada em combinação com a quercetina e rutina em C. neoformans ATCC e reduzida quando combinados com rutina em um isolado clínico de C. neoformans. Além disso, a combinação de quercetina com anfotericina B pode reduzir a toxicidade da droga em eritrócitos. Os resultados sugerem que quercetina e rutina são potenciais agentes para combinação com anfotericina B, a fim de reduzir a dose de anfotericina, diminuindo os efeitos colaterais e melhorando sua eficácia antifúngica.

Antifungal activity of propolis against Candidaspecies isolated from cases of chronic periodontitis

This research evaluated the fungistatic and fungicidal activities of red propolis alcoholic extract (RPAE) against different Candida species isolated from chronic periodontitis cases, and compared with chlorhexidine (CHX). Nineteen samples of Candida species (C. albicans [n = 12], C. tropicalis [n = 5] andC. glabrata[n = 2]) isolated from chronic periodontitis cases were analyzed. The fungistatic and fungicidal activity of both RPAE and CHX were evaluated using fluconazole and C. parapsilosis (ATCC 6258) as a control. Fungistatic activity was analyzed based on the Clinical and Laboratory Standards Institute (CLSI) reference procedure to determine the minimum inhibitory concentrations. Fungicidal activity was established according to the absence of fungal growth on Sabouraud Dextrose Agar medium. The fungistatic and fungicidal activities of RPAE were observed, respectively, at 32-64 μg/mL and 64-512 μg/mL for C.albicans, 64 μg/mL and 64-256 μg/mL for C. glabrata, and 32-64 μg/mL and 64 µg/mL for C. tropicalis. CHX fungistatic activity was observed at concentrations of 0.003-1.92 µg/mL for C. albicans, 1.92 µg/mL for C. glabrata, and 0.03-1.92 µg/mL for C. tropicalis. Fluconazole fungistatic activity ranged between 1-64 μg/mL, and fungicidal activity occurred at 8-64 μg/mL, for the threeCandida species analyzed. All the Candidaspecies were susceptible to RPAE antifungal activity, but five samples ofC.albicans, one ofC.tropicalis and one ofC.glabrata were resistant to fluconazole antifungal activity. CHX showed fungistatic activity against all the Candida species analyzed. The antifungal potential of these substances suggests that they can be applied as an alternative treatment for diseases affected by these species.

Use of mueller-iinton broth and agar in the germ tue test / Utilização de ágar e caldo Mueller-Hinton na prova do tubo germinativo

Candida albicans is often isolated from clinical samples, thus its presumptive differentiation from other species of the same genus can be based on its ability to form the germ tube in human serum. Nevertheless, there are two other species that share this characteristic: C. dubliniensis and C. africana. The aim of this study was to compare four different substrates to perform the germ tube (GT) test. The Candida spp. isolates were identified using a manual system (135 C. albicans, 24 C. tropicalis and one C. dubliniensis). The germ tube test was performed with fresh, previously frozen serum and Mueller-Hinton (MH) broth and agar. GT was observed in 96% (130/136) of the isolates through the fresh serum technique, 94% (128/136) through previously frozen serum, 92% (125/136) in MH agar, and 90% (122/136) in MH broth. The sensitivity of each test was higher than 90%, with 100% specificity. Both the MH agar and broth were able to identify the true positives, and false positives were not found. However, some C. albicans isolates were not identified. MH agar and broth may be used in laboratory for the rapid presumptive identification of C. albicans, as an alternative method for germ tube test.

Candida albicans é frequentemente isolada em amostras clínicas, assim a sua diferenciação presuntiva de outras espécies do gênero pode ser baseada na habilidade em formar o tubo germinativo em soro humano. Entretanto, existem outras duas espécies que também possuem essa característica, C. dubliniensis e C. africana. O objetivo foi comparar quatro diferentes substratos para a realização da prova do tubo germinativo (TG). Utilizou-se isolados de Candida spp. identificados através de meio manual (135 C. albicans, 24 C. tropicalis e um C. dubliniensis). A prova do tubo germinativo foi realizada utilizando soro previamente congelado e fresco, caldo e ágar Mueller-Hinton (MH). O TG através da técnica do soro a fresco foi observado em 96% (130/136), 94% (128/136) através do soro previamente congelado, 92% (125/136) no ágar e 90% (122/136) no caldo MH. A sensibilidade de cada teste foi maior que 90% e especificidade de 100%. Tanto o caldo quanto o ágar MH foram capazes de identificar apenas os verdadeiros positivos e não ocorrendo falsos positivos, porém deixaram de identificar alguns isolados de C. albicans. O ágar e o caldo MH podem ser utilizados na rápida e presuntiva identificação laboratorial de C. albicans, como uma alternativa para o teste do tubo germinativo.

New cultive medium for bioconversion of C5 fraction from sugarcane bagasse using rice bran extract

The use of hemicellulosic hydrolysates in bioprocesses requires supplementation as to ensure the best fermentative performance of microorganisms. However, in light of conflicting data in the literature, it is necessary to establish an inexpensive and applicable medium for the development of bioprocesses. This paper evaluates the fermentative performance of Scheffersomyces (Pichia) stipitis and Candida guilliermondii growth in sugarcane bagasse hemicellulosic hydrolysate supplemented with different nitrogen sources including rice bran extract, an important by-product of agroindustry and source of vitamins and amino acids. Experiments were carried out with hydrolysate supplemented with rice bran extract and (NH4)2SO4; peptone and yeast extract; (NH4)2SO4, peptone and yeast extract and non-supplemented hydrolysate as a control. S. stipitis produced only ethanol, while C. guilliermondii produced xylitol as the main product and ethanol as by-product. Maximum ethanol production by S. stipitis was observed when sugarcane bagasse hemicellulosic hydrolysate was supplemented with (NH4)2SO4, peptone and yeast extract. Differently, the maximum xylitol formation by C. guilliermondii was obtained by employing hydrolysate supplemented with (NH4)2SO4 and rice bran extract. Together, these findings indicate that: a) for both yeasts (NH4)2SO4 was required as an inorganic nitrogen source to supplement sugarcane bagasse hydrolysate; b) for S. stipitis, sugarcane hemicellulosic hydrolysate must be supplemented with peptone and yeast extract as organic nitrogen source; and: c) for C. guilliermondii, it must be supplemented with rice bran extract. The present study designed a fermentation medium employing hemicellulosic hydrolysate and provides a basis for studies about value-added products as ethanol and xylitol from lignocellulosic materials.

Production and characterization of an extracellular lipase from Candida guilliermondii

Extracellular lipases from the endophytic yeast Candida guilliermondii isolated from castor leaves (Ricinus communis L.) were produced using low-cost raw materials such as agro-industrial residues and applying them in the esterification of oleic acid for evaluating their potential use in biodiesel production. After partial purification using ammonium sulfate, the enzyme was characterized and presented higher activity (26.8 ± 1.5 U mL-1) in the presence of 5 mmol L-1 NaCl at 30 ºC and pH 6.5. The production through submerged fermentation was formerly performed in 150 mL erlenmeyer flasks and, once the enzyme production was verified, assays in a 14 L bioreactor were conducted, obtaining 18 ± 1.4 U mL-1. The produced enzyme was applied in the oleic acid esterification under different solvents: hexane, cyclohexane or cyclohexanone) and different acid:alcohol molar ratios. Higher ester conversion rate (81%) was obtained using hexane and the molar ratio of 1:9 was the best conditions using methanol. The results suggest the potential for development of endophytic yeast in the production of biocatalyst through submerged fermentation using agroindustrial residues as culture medium.

Queijo minas frescal suplementado com probiótico para o controle de Candida sp. isolados da cavidade bucal de usuários de próteses totais - estudo duplo-cego randomizado / Minas frescal cheese supplemented with probiotics for control of Candida sp. isolated from oral cavity of complete denture wearers - double-blind randomized trial

Os probióticos são organismos vivos que quando ingeridos em quantidades adequadas, conferem benefícios aos indivíduos. Um destes benefícios é a capacidade de inibir o crescimento de micro-organismos patogênicos como Candida. Para avaliar a redução ou inibição de Candida em usuários de próteses totais promovida pela ingestão de probióticos, foram recrutados 135 pacientes, dos quais 59 permaneceram até o final do período experimental de 8 semanas. A ingestão dos probióticos ocorreu através de sua incorporação em queijo minas frescal. Foram selecionados pacientes usuários de próteses totais, uni ou bimaxilares albergando Candida na cavidade bucal. As amostras foram coletadas através de um enxaguado bucal 1 semana antes do início do experimento e ao final do período experimental de 8 semanas. Foi determinado o número de UFC de Candida/ml da amostra. Os sujeitos da pesquisa (n=59) receberam embalagens com 20g de queijo minas frescal a cada 14 dias por 8 semanas consecutivas. Os participantes foram divididos aleatoriamente em três grupos, experimental 1 e 2, e controle. Os grupos experimentais receberam os queijos suplementados com probióticos, respectivamente, Lactobacillus acidophilus NCFM e Lactobacillus rhamnosus Lr-32, e o grupo controle recebeu o queijo sem suplementação probiótica. Os queijos possuíam semelhança quanto a aroma e cor. Além da quantificação de Candida, foram realizados testes para identificação das espécies e avaliação da atividade enzimática da fosfolipase e proteinase. Os resultados inferem que os probióticos L. acidophilus NCFM e L. rhamnosus Lr-32 suplementados em queijo minas frescal foram capazes de reduzir os níveis de Candida bucal, em pacientes usuários de próteses totais, após ingestão diária dos queijos por 8 semanas consecutivas. Antes e após 8 semanas do experimento, Candida albicans foi a espécie mais isolada dos pacientes nos três grupos avaliados...

Probiotics are living organisms which when ingested in adequate amounts confer benefits to individuals. One of these benefits is the ability to inhibit the growth of pathogenic microorganisms such as Candida. Reduction or inhibition of Candida isolated from complete denture wearers, uni or bimaxillary, was evaluated by ingestion of minas frescal cheese supplemented with probiotics. 135 patients were recruited, who 59 remained until the end of the experiment. The participants were patients in treatment at the Faculdade de Odontologia da Universidade de São Paulo. The samples were collected through a rinsed mouthwash at the base line and 8 weeks of experiment, before the installation of the new prosthesis. The samples were inoculated on Saubouraud agar dextrose with chloramphenicol plates, incubated for 24-48 hours, in order to quantify in colony-forming units per milliliter (CFU/mL) of Candida presents in the oral cavity of these patients. Subjects who presented fungus positive culture received packages containing 20g of minas frescal cheese every 14 days for 8 consecutive weeks. The participants were divided into three groups: two experimental and one control. The experimental groups received the cheeses supplemented with probiotics Lactobacillus acidophilus NCFM and Lactobacillus rhamnosus Lr-32. The control group received the cheese without probiotic supplementation. The cheeses had similarity as flavoring and coloring. In addition to the quantification of Candida, tests for species identification, phospholipase and proteinase activities were carried out. The results infer that L. acidophilus NCFM and L. rhamnosus Lr-32 supplemented in minas frescal cheese were able to reduce the levels of oral Candida in complete denture wearers, after daily intake of cheeses for 8 weeks of the experiment. At baseline and after 8 weeks of the experiment Candida albicans was the most frequently isolated from denture wearers in all groups...

Evaluation of chromogenic media and seminested PCR in the identification of Candida species

Identification of Candida cultured from various clinical specimens to the species level is increasingly necessary for clinical laboratories. Although sn PCR identifies the species within hours but its cost-effectiveness is to be considered. So there is always a need for media which help in the isolation and identification at the species level. The study aimed to evaluate the performance of different chromogenic media and to compare the effectiveness of the traditional phenotypic methods vs. seminested polymerase chain reaction (sn PCR) for identification of Candida species. One hundred and twenty seven Candida strains isolated from various clinical specimens were identified by conventional methods, four different chromogenic media and sn PCR. HiCrome Candida Differential and CHROMagar Candida media showed comparably high sensitivities and specificities in the identification of C. albicans, C. tropicalis, C. glabrata and C. krusei. CHROMagar Candida had an extra advantage of identifying all C. parapsilosis isolates. CHROMagar-Pal's medium identified C. albicans, C. tropicalis and C. krusei with high sensitivities and specificities, but couldn't identify C. glabrata or C. parapsilosis. It was the only medium that identified C. dubliniensis with a sensitivity and specificity of 100%. Biggy agar showed the least sensitivities and specificities. The overall concordance of the snPCR compared to the conventional tests including CHROMAgar Candida in the identification of Candida species was 97.5%. The use of CHROMAgar Candida medium is an easy and accurate method for presumptive identification of the most commonly encountered Candida spp.

In vitro evaluation of vincristine and fluconazole combination against Candida

Infections associated with cancer are a major scourge and cause of substantial morbidity and mortality in cancer patients. The aim of present study was to appraise the in vitro activity of anticancer agent vincristine and antifungal fluconazole alone and in combination against Candida spp. Results were interpreted in terms of fractional inhibitory concentration index [FICI]. Antifungal activity of fluconazole showed marked synergism when used in combination with vincristine, with FICI ranging from 0.25-0.5 against different Candida spp. Although, the use of vincristine with fluconazole is always disputed due to its side effects including decreased peristalsis, but the present research can help to perform suitability analysis of fluconazole use in life threatening invasive candidiasis associated with cancer patients. In addition, the synergism in antifungal activity after using with vincristine also warrants further research in the direction of minimizing adverse reaction associated with combined use of fluconazole and vincristine

Onicomicosis: epidemiología, agentes causales y evaluación de los métodos diagnósticos de laboratorio / Onychomycoses: epidemiology, causative agents and assessment of diagnostic laboratory methods

Desde marzo de 2007 hasta marzo de 2011 se estudiaron prospectivamente 414 pacientes con onicodistrofias en un laboratorio privado de Esquel. La prevalencia de onicomicosis de pie fue del 78 %; la de mano, del 58 %. Los principales agentes etiológicos fueron Trichophyton rubrum, Candida spp. y Trichophyton mentagrophytes. El desarrollo de dermatofitos prevaleció en las onicopatías de pie y el de Candida spp. en las de uñas de mano (ambos, p < 0,05). En las onicomicosis candidiásicas predominaron especies diferentes a Candida albicans. Las onicomicosis fueron más frecuentes en los hombres que en las mujeres. A su vez, en los hombres hubo más aislamientos de T. rubrum en pies (p < 0,05) y mayor proporción de exámenes directos (ED) y cultivos positivos (ambos, p < 0,05). La correlación entre los resultados del ED y del cultivo fue del 68 %. El rédito de ambos métodos se asoció a un mayor tamaño de la lesión ungueal. El ED fue más efectivo en onicodistrofias que superaban los 5 años de evolución. La positividad del cultivo fue independiente de la cronicidad de la onicodistrofia.

Since March 2007 to March 2011, 414 patients with onychopathies were prospectively analyzed. Prevalence of the toenail and fingernail mycoses was 78 % and 58 %, respectively. The major etiological agents were Trichophyton rubrum, Candida spp. and Trichophyton mentagrophytes. Dermatophytes were more frequently cultured from toenails, whereas Candida spp. from fingernails (both, p < 0.05). In candidal onychomycosis, species different from C. albicans were prevalent. A higher prevalence of toenail and fingernail mycoses, a predominance of T. rubrum in toenails (p < 0.05), and greater positivity in the direct examination (DE) and in culture (both, p < 0.05) were more frequently observed in men than in women. The correlation between DE and culture was 68 %. DE and culture yields were associated with a greater size lesion. DE was more effective in onycodystrophies with duration of more than 5 years. Culture positivity was independent of nail affection chronicity.

Nasal carriage of Staphylococcus aureus and Candida species in immunocompetent individuals

El propósito de este trabajo fue analizar la prevalencia de Staphylococcus aureus y especies de Candida en muestras demucosa nasal de 100 individuos inmunocompetentes de ambos sexos con edades entre 18-70 años, durante el examen clínico estomatológico. Las muestras fueron obtenidas con hisoposestériles sobre la mucosa de ambas fosas nasales. Se realizó observación en fresco, tinción de Gram, Giemsa y cultivos en medios selectivos y diferenciales a 37ºC para el aislamiento e identificación de los microorganismos seleccionados, pruebas bioquímicas convencionales y equipos comerciales y estudios moleculares mediante la prueba de PCR. Con un termo higrómetro digital se midió la temperatura ambiente cuyo promedio en el momento de la toma en el consultoriofue de 25±2 oC y la humedad relativa ambiente fue del 66±11 por ciento.S aureus se aisló en el 38 por ciento de las muestras y dentro del mismo, 4 por ciento fueron meticilino resistentes (MRSA) siendo genéticamente 2 por ciento de la Comunidad (MRSA-CA) y 2 por ciento Hospitalarios (MRSA-HA).En el 23 por ciento de las muestras fue identificada Candida siendo la especie prevalente C. albicans: 19 por ciento y en menor proporción C. dubliniensis: 3 opr ciento , C. krusei: 1 por ciento. Se registró una asociación significativa entre Candida y S. aureus (Chi-cuadrado=27,75; gl=1; (p<0,001). La cavidad nasal constituye un reservorio yla identificación de género y especie contribuye a la adecuada vigilancia epidemiológica.

Microbial colonization in orthodontic mini-implants

Peri-implant inflammation contributes for loss of secondary stability of orthodontic mini-implants. The investigation of microbial colonization in this area would benefit its control, and consequently favor the long-term success of mini-implants. Therefore, the aim of this study was to determine the establishment and the evolution of microbial colonization process in orthodontic mini-implants for 3 months, since the time of their installation. One-hundred and fifty samples collected from 15 mini-implants were investigated from baseline up to 3 months. The biological material was obtained from peri-implant area using paper points. Nonspecific, Streptococcus spp, Lactobacillus casei and Candida spp colonizations were analyzed by cell growth methods. Porphyromonas gingivalis colonization was observed by 16S rDNA-directed polymerase chain reaction. Data from cell growth were submitted to the Wilcoxon sign rank test and results from molecular analysis were presented in a descriptive way. There was no significant difference in the microbial colonization among the examined time intervals, except for Streptococcus spp, between baseline and 24 h, which characterized the initial colonization in this time interval. Lactobacillus casei and Candida spp colonizations were insignificant. No Porphyromonas gingivalis was detected among the analyzed samples. The microbial colonization of mini-implants did not significantly change during the study. However, it should be monitored by orthodontists, since it is an important factor for mini-implants success.

A inflamação peri-implantar contribui para a perda da estabilidade secundária dos mini-implantes ortodônticos. A investigação da colonização microbiana desta área beneficiaria o seu controle e, consequentemente, favoreceria o sucesso dos mini-implantes a longo prazo. Portanto, o objetivo dos autores foi determinar o estabelecimento e evolução do processo de colonização microbiana em mini-implantes ortodônticos por três meses desde a instalação. Cento e cinquenta amostras coletadas de 15 mini-implantes foram investigadas desde o tempo inicial até 3 meses. O material biológico foi obtido da área peri-implantar com auxílio de cones de papel absorvente. As colonizações inespecíficas de Streptococcus spp, Lactobacillus casei e Candida spp foram analisadas por métodos de crescimento celular. A colonização por Porphyromonas gingivalis foi observada por meio da reação em cadeia da polimerase 16S rDNA. Os dados do crescimento celular foram submetidos ao teste de Wilcoxon sign rank e os resultados da biologia molecular foram apresentados de modo descritivo. Não houve diferença estatisticamente significante da colonização microbiana entre os intervalos de tempo avaliados, exceto para Streptococcus spp entre os tempos inicial e 24 h, o que caracterizou o início da colonização neste intervalo de tempo. As colonizações por Lactobacillus casei e Candida spp foram insignificantes. Não foi detectada a presença de Porphyromonas gingivalis nas amostras analisadas. A colonização microbiana nos mini-implantes não se alterou significativamente durante o estudo. No entanto, deve ser monitorada por ortodontistas, uma vez que é um fator importante para o sucesso dos mini-implantes.

Differentiation between Candida albicans and Candida dubliniensis using hypertonic Sabouraud broth and tobacco agar / Diferenciação entre Candida albicans e Candida dubliniensis usando caldo Sabouraud hipertônico e ágar Tabaco

INTRODUCTION: Opportunistic fungal infections in immunocompromised hosts are caused by Candida species, and the majority of such infections are due to Candida albicans. However, the emerging pathogen Candida dubliniensis demonstrates several phenotypic characteristics in common with C. albicans, such as production of germ tubes and chlamydospores, calling attention to the development of stable resistance to fluconazole in vitro. The aim of this study was to evaluate the performance of biochemistry identification in the differentiating between C. albicans and C. dubliniensis, by phenotyping of yeast identified as C. albicans. METHODS: Seventy-nine isolates identified as C. albicans by the API system ID 32C were grown on Sabouraud dextrose agar at 30°C for 24-48h and then inoculated on hypertonic Sabouraud broth and tobacco agar. RESULTS: Our results showed that 17 (21.5%) isolates were growth-inhibited on hypertonic Sabouraud broth, a phenotypic trait inconsistent with C. albicans in this medium. However, the results observed on tobacco agar showed that only 9 (11.4%) of the growth-inhibited isolates produced characteristic colonies of C. dubliniensis (rough colonies, yellowish-brown with abundant fragments of hyphae and chlamydospores). CONCLUSIONS: The results suggest that this method is a simple tool for screening C. albicans and non-albicans yeast and for verification of automated identification.

INTRODUÇÃO: Infecções fúngicas oportunistas em hospedeiros imunocomprometidos são causadas por espécies de Candida, cuja maioria das infecções se deve a Candida albicans. Entretanto, o patógeno emergente Candida dubliniensis demonstra várias características fenotípicas em comum com C. albicans, tais como produção de tubo germinativo e clamidósporos, solicitando atenção por desenvolver resistência in vitro estável ao fluconazol. O objetivo do presente estudo foi avaliar a performance da identificação bioquímica na diferenciação entre C. albicans e Candida dubliniensis, analisando fenotipicamente leveduras previamente identificadas como C. albicans. MÉTODOS: Setenta e oito isolados identificados como C. albicans pelo sistema API ID 32C foram cultivados em ágar Sabouraud dextrose a 30°C por 24-48h e em seguida inoculados em caldo hipertônico Sabouraud e agar tabaco. RESULTADOS: Nossos resultados mostraram que 17 (21,5%) isolados tiveram o crescimento inibido no caldo hipertônico Sabouraud, característica fenotípica inconsistente para C. albicans neste meio de cultura. Entretanto, os resultados observados em ágar tabaco mostraram que somente 9 (11,4%) dos isolados inibidos produziram colônias características de C. dubliniensis (colônias rugosas, marrom-amarelada com fragmentos de hifas e abundantes clamidósporos). CONCLUSÕES: Os resultados obtidos sugerem que este é um instrumento simples para triagem entre leveduras de C. albicans e não-albicans, bem como confirmação de identificação automatizada.

Productividad y selectividad del medio de cultivo a partir de guayaba agria (psidium araca) en el crecimiento de levaduras nativas del género candida sp / Productivity and selectivity of culture medium from the sour guava (psidium araca) in the growth of native yeast candida sp

El presente trabajo se llevó a cabo para evaluar la eficiencia del medio de cultivo a partir de guayaba agria (Psidium araca) frente a medios comerciales en el crecimiento de tres cepas nativas: Candida guillermondii, Candida famita y Candida sp. Se evaluó el crecimiento microbiano a diferentes concentraciones de fruta, 5, 10, 25 y 50% p/v, tomando como control los medios comerciales: Malta, Sabouraud y agar papa dextrosa (PDA). La productividad y selectividad del medio de guayaba agria fue determinada mediante el método Ecométrico en un tiempo de 48 horas. Los análisis estadísticos aplicados para evaluar y comparar el crecimiento de las cepas en los medios comerciales y en el medio de guayaba agria a diferentes concentraciones demostraron lo siguiente: Candida guillermondii presentó crecimiento mayor o igual a 25 y 50% p/v comparado con los medios comerciales; Candida famata y Candida sp presentaron mejores crecimientos al 5% p/v, con respecto a los diferentes medios comerciales. Los resultados demostraron que el medio de cultivo es altamente productivo y no selectivo, lo que representa una alternativa en la conservación, el mantenimiento y el desarrollo de las levaduras estudiadas.

This work was carried out to evaluate the efficiency of the culture medium from sour guava (Psidium araca) against commercial media in the growth of three native strains: Candida guillermondii, Candida famata and Candida sp. Microbial growth was evaluated at different concentrations of fruit, 5, 10, 25, 50% w /v, using as control the commercial media: Malta, Sabouraud and PDA (Potato Dextrose Agar). The productivity and selectivity of the sour guava medium was determined by the Ecometric method in a time of 48 hours. The applied statistical analysis to evaluate and compare growth of strains in commercial culture medium and in the medium from sour guava at different concentrations showed: Candida guillermondii grew greater than or equal to 25 and 50% w / v compared with commercial medium, Candida famata and Candida sp showed better growth at 5% w / v, with respect to commercial medium. The results showed that the medium is highly productive and non-selective representing an alternative to the conservation, maintenance and development of the yeasts.

Long-term efficacy of denture cleansers in preventing Candida spp. biofilm recolonization on liner surface

This study evaluated the long-term efficacy of denture cleansers against Candida spp. biofilm recolonization on liner surface. Specimens were fabricated of a poly(methyl methacrylate)-based denture liner and had their surface roughness evaluated at baseline and after cleansing treatments. C. albicans or C. glabrata biofilms were formed on liner surface for 48 h, and then the specimens were randomly assigned to one of cleaning treatments: two alkaline peroxides (soaking for 3 or 15 min), 0.5 percent sodium hypochlorite (10 min) or distilled water (control; 15 min). After the treatments, the specimens were sonicated to disrupt the biofilm, and residual cells were counted (cell/mL). Long-term effectiveness of the cleaning processes was determined by submitting a set of cleaned specimens to biofilm growth conditions for 48 h followed by estimation of cell counts. The topography of specimens after cleaning treatments was analyzed by SEM. Data were analyzed by ANOVA and Tukey's test (α; = 0.05). Results of cell count estimation showed significant differences in cleanliness among the treatments (p < 0.001), and it could be observed by SEM. However, no significant difference (p > 0.05) was observed among the Candida species regarding the recolonization condition. Alkaline denture cleansers showed similar cleaning performance and both differed from the control (p < 0.001). Sodium hypochlorite was the only treatment that removed biofilm efficiently, since no viable cells were found after its use. In conclusion, alkaline peroxide denture cleansers were not effective in removing Candida spp. biofilm from denture liner surfaces and preventing biofilm recolonization.

Identification of candida spp. by phenotypic tests and PCR

The correct identification of Candida species is of great importance, as it presents prognostic and therapeutical significance, allowing an early and appropriate antifungical therapy. The purpose of this study was to identify isolates of Candida spp. from oral mucosa of 38 patients with oral candidosis evaluated in 2004 by phenotypic methods and PCR, discriminating C. albicans from the other Candida species. The tests used for phenotypic analysis were germ-tube and chlamydoconidia production, culture in CHROMAgarTM Candida, carbohydrate assimilation test, growth at 45ºC and culture in Tween 80 agar. Genotypic confirmation was performed by PCR. Phenotypic tests showed that 63.2 percent strains formed germ-tubes, 73.7 percent produced chlamydoconidia, and 63.2 percent showed green colonies in chromogenic medium, presumptively indicating C. albicans or C. dubliniensis. The carbohydrate assimilation test confirmed these results. A total of 21 percent strains were identified as C. krusei and 13.2 percent were indicative of C. tropicalis. Of these later strains, three produced chlamydoconidia. The association of other phenotypic tests with culture in Tween 80 agar identified 95.8 percent of strains as C. albicans and 4.2 percent as C. dubliniensis. All 24 strains indicative of C. albicans and C. dubliniensis were confirmed by PCR as C. albicans.

Use of Pseudomonas stutzeri and Candida utilis in the improvement of the conditions of Artemia culture and protection against pathogens

To evaluate the effect of two bacterial strains isolated from Artemia cysts and yeast (Candida utilis) on the survival, growth and total biomass production of its larvae, challenge tests were performed with Candida utilis, Pseudomonas stutzeri and Pasteurella haemolityca. In addition, a pathogenic strain of Vibrio alginolyticus was tested for comparative purposes. Pseudomonas stutzeri and Candida utilis have no impact on survival, but enhance growth and total biomass production of the larvae. However, we noted that Pasteurella haemolityca affect negatively Artemia larvae. The adhesion and antagonism assay demonstrates that Candida utilis and Pseudomonas stutzeri are fairly adherent and play an important role in the enhancement of the protection of Artemia culture against pathogens. On the basis of these results, it's suggested that it's possible to use Candida utilis and Pseudomonas stutzeri, potential candidates, as probiotic for the culture of Artemia larvae.