As infecções causadas por espécies de Candida são problema de grande impacto para a saúde pública, devido à alta incidência em pacientes hospitalizados e como causa de mortalidade. O presente estudo teve como objetivo avaliar a frequência de Candida spp. isoladas de pacientes hospitalizados, assim como a sensibilidade aos antifúngicos e o polimorfismo genético por RAPD-PCR. Os microrganismos incluíram isolados de hemocultura, líquido abdominal e ponta de cateter venoso central de pacientes internados no Hospital de Clínicas da Universidade Federal de Uberlândia, região do Triângulo Mineiro, Minas Gerais, Brasil, no período de julho de 2010-junho de 2011. Os testes de sensibilidade aos antifúngicos foram realizados por microdiluição em caldo e na análise por RAPD-PCR foram utilizados os oligonucleotídeos OPA09, OPB11, e OPE06. Dos 63 isolados, 18 (28,5%) foram C. albicans, 20 (31,7%) C. parapsilosis, 14 (22,2%) C. tropicalis, quatro (6,4%) C. glabrata, quatro (6,4%) C. krusei, dois (3,3%) C. kefyr, e um (1,6%) C. lusitaniae. Resistência in-vitro à anfotericina B foi observada em 12,7% dos isolados. Não foi observada resistência in-vitro aos azólicos, exceto para os isolados de C. krusei. Os oligonucleotídeos OPA09 e OPB11 possibilitaram distinguir diferentes espécies. Isolados de C. albicans apresentaram seis clusters e o complexo C. parapsilosis, cinco clusters, com o iniciador OPA09, por RAPD-PCR, mostrando a variabilidade genética daquelas espécies. Conclui-se que o complexo C. parapsilosis foi a espécie mais frequente, e a maioria dos isolados foi sensível in vitro aos antifúngicos testados. Alto polimorfismo genético foi observado para os isolados de C. albicanse complexo C. parapsilosis, principalmente com o oligonucleotídeo OPA09.

Infections by Candida species are a high-impact problem in public health due to their wide incidence in hospitalized patients. The goal of this study was to evaluate frequency, susceptibility to antifungals, and genetic polymorphism of Candida species isolated from clinical specimens of hospitalized patients. The Candida isolates included in this study were obtained from blood cultures, abdominal fluids, and central venous catheters (CVC) of hospitalized patients at the Clinical Hospital of the Federal University of Uberlândia during the period of July 2010 - June 2011. Susceptibility tests were conducted by the broth microdilution method. The RAPD-PCR tests used employed initiator oligonucleotides OPA09, OPB11, and OPE06. Of the 63 Candida isolates, 18 (28.5%) were C. albicans, 20 (31.7%) were C. parapsilosis complex species, 14 (22.2%) C. tropicalis, four (6.4%) C. glabrata, four (6.4%) C. krusei, two (3.3%) C. kefyr, and one (1.6%) C. lusitaniae. In vitro resistance to amphotericin B was observed in 12.7% of isolates. In vitroresistance to azoles was not detected, except for C. krusei. The two primers, OPA09 and OPB11, were able to distinguish different species. Isolates of C. albicans and C. parapsilosis complex species presented six and five clusters, respectively, with the OPA09 marker by RAPD-PCR, showing the genetic variability of the isolates of those species. It was concluded that members of the C. parapsilosis complex were the most frequent species found, and most isolates were susceptible to the antifungals amphotericin B, flucozanole, and itraconazole. High genetic polymorphisms were observed for isolates of C. albicans and C. parapsilosis complex species, mainly with the OPA09 marker.

Various organisms have been characterized by molecular methods, including fungi of the genus Cryptococcus. The purposes of this study were: to determine the discriminatory potential of the RAPD (Random Amplified Polymorphic DNA) primers, the pattern of similarity of the Cryptococcus species, and discuss their useful application in epidemiological studies. We analyzed 10 isolates of each specie/group: C. albidus, C. laurentii complex, C. neoformans var. grubii, all from environmental source, and two ATCC strains, C. neoformans var. grubii ATCC 90112, and C. neoformans var. neoformans ATCC 28957 by RAPD-PCR using the primers CAV1, CAV2, ZAP19, ZAP20, OPB11 and SEQ6. The primers showed a good discriminatory power, revealing important differences between them and between species; the SEQ6 primer discriminated a larger number of isolates of three species. Isolates of C. laurentii showed greater genetic diversity than other species revealed by all six primers. Isolates of C. neoformans were more homogeneous. Only the primer CAV2 showed no amplification of DNA bands for C. albidus. It was concluded that the use of limited number of carefully selected primers allowed the discrimination of different isolates, and some primers (e.g., CAV2 for C. albidus) may not to be applied to some species.

Foi avaliada a suscetibilidade in vitro de dermatófitos aos antifúngicos itraconazol, fluconazol e cetoconazol, pelos métodos macro e microdiluição em caldo, de acordo com as recomendações do CLSI, com algumas modificações. Foram estudados 20 isolados clínicos de lesões de unha e pele, sendo quatro Trichophyton mentagrophytes e 16 T. rubrum. A concentração inibitória mínima (CIM) para itraconazol variou de < 0,03 a 0,25 µg/mL pelo método da macrodiluição, e de < 0,03 a 0,5 µg/mL pela microdiluição em caldo; de 0,5 a 64 µg/mL e de 0,125 a 16 µg/mL para fluconazol, respectivamente, pela macro e microdiluição; e de < 0,03 a 0,5 µg/mL por ambos os métodos para cetoconazol. A concordância entre os dois métodos (considerando ± uma diluição) foi de 70% para itraconazol, 45% para fluconazol e 85% para cetoconazol. Conclui-se que os isolados estudados foram inibidos por concentrações relativamente baixas dos antifúngicos testados, e os dois métodos apresentam boa concordância, especialmente para itraconazol e cetoconazol.

The in vitro susceptibility of dermatophytes to the azole antifungals itraconazole, fluconazole and ketoconazole was evaluated by broth macro and microdilution methods, according to recommendations of the CLSI, with some adaptations. Twenty nail and skin clinical isolates, four of Trichophyton mentagrophytes and 16 of T. rubrum were selected for the tests. Itraconazole minimal inhibitory concentrations (MIC) varied from < 0.03 to 0.25 µg/mL in the macrodilution and from < 0.03 to 0.5 µg/mL in the microdilution methods; for fluconazole, MICs were in the ranges of 0.5 to 64 µg/mL and 0.125 to 16 µg/mL by the macro and microdilution methods, respectively, and from < 0.03 to 0.5 µg/mL by both methods for ketoconazole. Levels of agreement between the two methods (± one dilution) were 70% for itraconazole, 45% for fluconazole and 85% for ketoconazole. It is concluded that the strains selected were inhibited by relatively low concentrations of the antifungals tested and that the two methodologies are in good agreement especially for itraconazole and ketoconazole.

A capacidade de Cryptococcus spp produzir melanina em meios contendo compostos fenólicos é amplamente utilizada na identificação destas espécies no laboratório. O objetivo do presente trabalho foi comparar a produção desse pigmento em quatro meios de cultura por Cryptococcus sp. Foram testadas 16 cepas de Cryptococcus neoformans, 17 de Cryptococcus albidus, 13 de Cryptococcus laurentii, e 2 de Cryptococcus uniguttulatus nos meios: ágar batata e cenoura, ágar alpiste, ágar semente de girassol e ágar L-dopa. A produção de melanina foi avaliada com base na pigmentação das colônias, e demonstrada em 5 dias de incubação por 93,8% das cepas de Cryptococcus neoformans nos meios ágar batata e cenoura, ágar semente de girassol e ágar L-dopa. Dos isolados de Cryptococcus albidus, 29,4% produziram o pigmento em ágar batata e cenoura e L-dopa, 11,8% em ágar alpiste, e 36% em ágar girassol. De Cryptococcus laurentii, 53,8% produziram em batata e cenoura e em semente de girassol, 61,5% em L-dopa, 84,6% em ágar alpiste. Somente uma cepa de Cryptococcus uniguttulatus produziu fracamente o pigmento em ágar batata e cenoura.

The capacity of Cryptococcus spp to produce melanin in media containing phenol compounds is widely used for identifying these species in the laboratory. The aim of the present study was to compare the production of this pigment by Cryptococcus spp. in four culture media. Sixteen strains of Cryptococcus neoformans, 17 of Cryptococcus albidus, 13 of Cryptococcus laurentii and two of Cryptococcus uniguttulatus were tested in the following media: potato-carrot agar, Niger seed agar, sunflower seed agar and L-dopa agar. The melanin production was evaluated on the basis of colony pigmentation. Its production after five days of incubation was demonstrated by 93.8% of the strains of Cryptococcus neoformans in the media of potato-carrot agar, sunflower seed agar and L-dopa agar. From the isolates of Cryptococcus albidus, 29.4% produced the pigment in potato-carrot agar and L-dopa agar, 11.8% in Niger seed agar and 36% in sunflower seed agar. From Cryptococcus laurentii, 53.8% produced the pigment in potato-carrot agar and sunflower seed agar, 61.5% in L-dopa agar and 84.6% in Niger seed agar. Only one strain of Cryptococcus uniguttulatus presented slight production of the pigment, in potato-carrot agar.

Este trabalho teve como objetivo, avaliar a presença de dermatófitos, especificamente em unhas, pés e mãos de estudantes universitários com e sem lesões sugestivas de dermatofitose. Foram coletadas 280 amostras dessas regiões, das quais 31 (11,1%) apresentaram positividade apenas pelo exame direto, e 20 (7,1%) tiveram, além do exame direto positivo, crescimento de dermatófito, mediante cultivo da amostra biológica. T. rubrum foi o dermatófito isolado com maior freqüência (80%), seguido por T. mentagrophytes (20%). Considerando os sítios analisados neste trabalho, a ocorrência de dermatófitos foi observada em 10,4% nas unhas dos pés, 5% nas escamas de pés, 2,5% nas unhas das mãos e apenas 0,4% nas escamas das mãos.

The objective of this paper was to evaluate the occurrence of dermatophytes, specifically in the nails, feet and hands of university students with and without lesions. Two hundred and eighty samples were collected; 31 (11.1%) were positive by direct examination, while only 20 (7.1%) showed dermatophyte growth in culture, as well as direct positive examination. Trichophyton rubrum was the most frequently isolated (80%) dermatophyte followed by T. mentagrophytes (20%). Considering the sites analyzed, dermatophyte occurrence was: 10.4% in toenails, 5% in foot skin, 2.5% in fingernails and 0.4% in hand skin.

Infections by Cryptococcus strains other than C. neoformans have been detected in immunocompromised patients. Of these strains, three are considered human pathogens: C. albidus, C. laurenttii, and C. uniguttulatus. This study deals with the in vitro susceptibility of Cryptococcus to drugs such as amphotericin B, itraconazole, fluconazole, and 5-fluorocytosine. Environmental Cryptococcus isolates (50) distributed as follows: C. neoformans var. neoformans (16), C. albidus (17), C. laurentii (14), and C. uniguttulatus (3) were evaluated by the micro and macrodilution techniques, according to EUCAST and NCCLS recommendations, respectively. Considering both methodologies the respective minimal inhibitory concentrations (MIC) were 0.125 and 2 µg/ml for amphotericin B, 0.06 and 8 µg/ml for itraconazole, and 0.5 and more than 64 µg/ml for fluconazole and 5-fluorocytosine. Agreement percentages for the two methodologies were 100% for amphotericin B and fluconazole for all the strains tested. For itraconazole, the agreement percentage was 81.3% in the C. neoformans strain and 100% for all the others. All species had a agreement percentage of 94.1 to 100% when susceptibility to 5-fluorocytosine was tested. It is concluded that environmental isolates of C. neoformans var. neoformans, C. albidus, C. laurentii, and C. uniguttulatus may show high MICs against certain drugs, suggesting in vitro primary resistance to the antifungals tested.

Foram avaliadas quanto a produção de exoenzimas fosfolipase e proteinase, 79 amostras de Candida isoladas da cavidade bucal de pacientes com lesões bucais características de candidose e indivíduos com boca clinicamente normal, atendidos na Faculdade de Odontologia de Ribeirão Preto USP. Dentre as cepas de C. albicans isoladas de lesões bucais, a fosfolipase e proteinase foram detectadas em, respectivamente, 83,3% e 66,7%. C. tropicalis e C. parapsilosis produziram somente proteinase. Quanto às cepas isoladas de nichos sem lesão, do total de 32 C. albicans, 71,9% apresentaram fosfolipase e 68,7% proteinase. C. tropicalis apresentou apenas a enzima proteinase, C. glabrata, C. krusei, C. guilliermondii e Candida spp, não apresentaram nenhuma das exoenzimas. Entre as amostras de C. albicans de ambos os grupos, o enzimotipo 22 (fosfolipase positiva e proteinase fracamente positiva) foi prevalente. Enzimotipos diferentes foram detectados em amostras da mesma espécie provenientes de mesmo paciente.

The production of phospholipase and proteinase exoenzymes was evaluated in seventy nine samples of Candida isolated from the oral cavity of patients with oral lesions characteristic of candidosis and from individuals presenting a clinically normal mouth, attended at the University of Dentistry of Ribeirão Preto USP. Among the strains of C. albicans isolated from oral lesions, the phospholipase and proteinase were detected in 83.3% and 66.7%, respectively. C. tropicalis and C. parapsilosis produced only proteinase. Regarding the isolated strains from niches without lesions, out of a total of 32 C. albicans, 71.9% presented phospholipase and 68.7% proteinase. C. tropicalis only presented the enzyme proteinase, C. glabrata, C. krusei, C. guilliermondii and Candida spp did not present any of the exoenzymes. Among the samples of C. albicans from both groups, the enzymotype 22 (positive phospholipase and proteinase weakly positive), was prevalent. Different enzymotypes of the same species were detected in samples collected from the same patient.

Foram avaliados 50 indivíduos, de ambos os sexos e faixa etária média de 42,5 anos, da clínica de Semiologia da FORP-USP, objetivando-se isolar e identificar leveduras na cavidade bucal, com e sem lesão, e determinar a DIM dos produtos comerciais Própolis (Apis-Flora) e Periogard (Colgate) frente às cepas isoladas. Leveduras do gênero Candida foram detectadas na saliva de 9(47,4%) indivíduos com boca clinicamente sadia, 18/81,8% portadores de lesões bucais e de 4/44,4% pacientes com desvio de normalidade; sendo detectadas em 19/86,4% lesões. No grupo com candidose bucal, respectivamente de língua e lesão, isolou-se C.tropicalis (8% e 10,7%), C.glabrata 4% e 3,6%) e C.parapsilosis (2% e 3,6%), além de C.albicans (71,4% e 67,8%) como espécie única e prevalente. A contagem de ufc total/ml de saliva demonstrou um valor médio no grupo com candidose bucal (171,5% x 10(3)) maior do que no controle (72,6 x 10(3)) e portadores de anormalidade (8,3 x 10(3)). A maioria das cepas testes 67/70 (95,7%) foi sensível aos anti-sépticos, sendo que a Própolis apresentou uma DIM igual a 1:20 para 54/70 (77,1%) e, o Periogard uma DIM de 1:160 para 42/70 (60%) cepas de nichos sadios; semelhante ao obtido com cepas de lesões bucais. Resultados diferentes ocorreram, principalmente, entre espécies diferentes. Os resultados indicam a possibilidade de se empregar os anti-sépticos Própolis e Periogard (clorexidina), na prevenção e na terapêutica da candidose bucal.

Fifty individuals of both sexes aged on average 45.2 years were evaluated at the Semiology Clinic of FORP-USP in order to isolate and identify yeasts from the oral cavity, with and without lesions, and to determine the maximal inhibitory dilution (MID) of the commercial products Propolis (Apis-Flora) and Periogard (Colgate) against the strains isolated. Yeasts of the genus Candida were detected in the saliva of 9/19 (47.4%) individuals with a clinically healthy mouth, 18/22 (81.8%) of individuals with oral lesions, and in 4/9 (44.4%) of patients with deviation from normality, and were detected in 19/22 (86.4%) of the lesions. In the group with oral candidiasis, we isolated in tongue and lesion, respectively for each specie: C.tropicalis (8% and 10.7%), C.glabrata (4% and 3.6%) and C.parapsilosis (2% and 3.6%), in addition to C.albicans (71.4% and 67.8%) as the only species and the prevalent. The total cfu counts/ml saliva showed a higher mean value in the group with oral candidiasis (171.5% x 10(3)) than in the control group (72.6 x 10(3)) or the group with abnormalities (8.3 x 10(3)). Most of the test strains 67/70 (95.71%) were sensitive to the antiseptics, with Propolis presenting a MID of 1:20 for 54/70/77.1%), and Periogard a MID of 1:160 for 42/70 (60%) strains from healthy sites, results similar to those obtained with strains from oral lesions. Different results were mainly observed among different species. The results indicate the possibility of using the antiseptics Propolis and Periogard (chlorhexidine) for the prevention and treatment of oral candidiasis.