Molecular epidemiology of carbapenem-resistant Acinetobacter baumannii from southern Brazil / Epidemiologia molecular de Acinetobacter baumannii resistente aos carbapenêmicos provenientes do sul do Brasil / Epidemiología molecular de Acinetobacter baumannii resistente a carbapenem del sur de Brasil

Background and Objectives: carbapenem resistance in Acinetobacter baumannii has reached extremely high levels worldwide, and class D OXA-type carbapenemases are the main associated mechanism. This study aimed to assess the phenotypic and molecular profile of clinical carbapenem-resistant A. baumannii (CRAb) isolates from a southern Brazilian border region. Methods: A. baumannii species was identified by the presence of the blaOXA-51 gene, and the susceptibility profile was determined by broth microdilution. The main carbapenemases were investigated by PCR and the molecular typing was performed by PFGE. Results: during the study, a total of 36 CRAb were recovered, of which 85.7% were from respiratory tract samples from ICU patients. High level resistance to were found in contrast to 100% of susceptibility for polymyxin B. The blaOXA-23 gene was present in 34 isolates and was the only one detected other than blaOXA-51. Molecular typing revealed the presence of four clonal strains, two of them endemic during the period of the study. Conclusion: to the best of our knowledge, our study brings the first data about resistance profile in Acinetobacter in the western border of southern Brazil and make aware of endemic clones of CRAb-producing-OXA-23 in this region of state, contributing for the construction of the national epidemiologic scenario of CRAb.(AU)

Justificativa e Objetivos: a resistência aos carbapenêmicos em Acinetobacter baumannii atingiu níveis extremamente altos em todo o mundo, e as carbapenemases do tipo OXA classe D são o principal mecanismo associado. O objetivo deste estudo foi avaliar o perfil fenotípico e molecular de isolados clínicos de A. baumannii resistentes aos carbapenêmicos (CRAb) de uma região de fronteira do sul do Brasil. Métodos: a espécie A. baumannii foi identificada através da presença do gene blaOXA-51, e o perfil de sensibilidade foi determinado por microdiluição em caldo. As principais carbapenemases foram investigadas por PCR, e a tipagem dos isolados de CRAb foi realizada por PFGE. Resultados: durante o período do estudo, 36 CRAb foram recuperados, dos quais 85,7% foram provenientes de amostras do trato respiratório de pacientes de UTI. Uma elevada resistência a aminoglicosídeos e fluoroquinolonas foi encontrada em contraste com 100% de sensibilidade a polimixina B. O gene blaOXA-23 foi encontrado em 34 isolados e foi o único detectado além do blaOXA-51. A tipagem molecular revelou a presença de quatro linhagens clonais, duas delas endêmicas ao longo do período do estudo. Conclusão: nosso estudo traz os primeiros dados sobre o perfil de resistência em Acinetobacter na fronteira oeste do sul do Brasil e alerta para a presença de clones endêmicos de CRAb produtores de OXA-23 nessa região, contribuindo para a construção do cenário epidemiológico nacional de CRAb.(AU)

Justificación y Objetivos: la resistencia a carbapenémicos en Acinetobacter baumannii ha alcanzado niveles extremadamente altos en todo el mundo y las carbapenemases OXA de clase D son el principal mecanismo asociado. El objetivo de este estudio fue evaluar el perfil fenotípico y molecular de los aislados clínicos de A. baumannii resistentes a carbapenémicos (CRAb) de una región fronteriza en el sur de Brasil. Métodos: la especie A. baumannii se identificó a través de la presencia del gen blaOXA-51 y el perfil de sensibilidad se determinó por microdilución en caldo. Las principales carbapenemasas fueron investigadas por PCR y la tipificación se hizo con PFGE. Resultados: durante el período de estudio, se recuperaron 36 CRAb, 85,7% de muestras del tracto respiratorio de pacientes de la UCI. Se encontró una alta resistencia a los aminoglucósidos y las fluoroquinolonas en contraste con 100% de sensibilidad a polimixina B. El gen blaOXA-23 se encontró en 34 aislamientos y fue el único detectado además de blaOXA-51. La tipificación molecular reveló la presencia de cuatro cepas clonales, dos de ellas endémicas durante el período de estudio. Conclusiones: hasta donde sabemos, nuestro estudio trae los primeros datos sobre el perfil de resistencia en Acinetobacter en la frontera oeste del sur de Brasil y reconoce los clones endémicos de CRAb productores de OXA.(AU)

Antimicrobial susceptibility of Salmonella Gallinarum and Salmonella Pullorum isolated from ill poultry in Brazil / Suscetibilidade a antimicrobianos de Salmonella Gallinarum e Salmonella Pullorum isolados de aves doentes no Brasil

ABSTRACT: Salmonella Gallinarum (S. Gallinarum) and Salmonella Pullorum (S. Pullorum) are poultry host-specific, agents of fowl typhoid and pullorum disease, respectively. These biovars cause septicemic infections, resulting in high mortality. Outbreaks are frequently reported worldwide, causing losses due to the elimination of infected flocks and treatments. The use of antimicrobial agents is frequent in poultry farms to prevent or treat gastrointestinal infections. In the present research it was evaluated the antimicrobial susceptibility of 50 S. Gallinarum and S. Pullorum isolates, from outbreaks that occurred between 1987 to 1991 and 2006 to 2013. The comparison of the susceptibility profiles showed that all isolates were susceptible to β-lactams. All isolates from 1987-1991 were susceptible to all antibiotics tested except NAL and CIP (78%). The susceptibility profile of S. Gallinarum (2006 - 2013 period) was the following NAL (58%), CIP (63%), ENR (67%), TET (92%), FFC (96%) and SXT (96%). S. Pullorum isolates (2006 - 2013 period) showed the following susceptibility rates to NAL (65%), CIP (71%), ENR (94%) and TET (94%). All isolates were susceptible to β-lactams tested, however, resistance to quinolones and fluoroquinolones increased over time. Furthermore, low levels of resistance to other antibiotics were found in recent isolates, such as tetracyclines.

RESUMO: Salmonella Gallinarum (S. Gallinarum) e Salmonella Pullorum (S. Pullorum) são patógenos hospedeiro-específico de aves, agentes do tifo aviário e pulorose, respectivamente. Estes biovares causam infecções septicêmicas, resultando em alta mortalidade. Surtos são frequentemente relatados em diversos países, causando prejuízos devido à eliminação de lotes infectados e tratamentos. Agentes antimicrobianos são utilizados frequentemente em granjas avícolas para prevenir ou tratar infecções gastrointestinais. No presente trabalho, foi avaliada a susceptibilidade antimicrobiana de 50 isolados de S. Gallinarum e S. Pullorum, obtidos durante surtos que ocorreram entre 1987 a 1991 e entre 2006 a 2013. A comparação dos perfis de sensibilidade mostrou que todas as amostras são sensíveis aos β-lactâmicos. Todos os isolados de 1987-1991 foram sensíveis a todos os antibióticos testados, exceto NAL e CIP (78%). O perfil de susceptibilidade de S. Gallinarum (surtos de 2006 a 2013) foi NAL (58%), CIP (63%), ENR (67%), TET (92%), FFC (96%) e SXT (96%). Isolados de S. Pullorum (surtos de 2006 a 2013) apresentaram as seguintes taxas de sensibilidade: NAL (65%), CIP (71%), ENR (94%) e TET (94%). Todas as amostras foram sensíveis ao β-lactâmicos testados, no entanto, a resistência às quinolonas e fluoroquinolonas aumentou ao longo do tempo. Além disso, baixos níveis de resistência a outros antibióticos foram encontrados em isolados recentes, tais como as tetraciclinas.

FREQUENCY OF Candida SPECIES IN A TERTIARY CARE HOSPITAL IN TRIANGULO MINEIRO, MINAS GERAIS STATE, BRAZIL / Frequência de espécies de Candida em hospital terciário do Triângulo Mineiro, Minas Gerais, Brasil

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 vitro resistance 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.

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. albicans e complexo C. parapsilosis, principalmente com o oligonucleotídeo OPA09.

Prior oropharyngeal colonization and ventilator-associated pneumonia

This study evaluated the relationship between previous colonization of the oropharynx and development of ventilator-associated pneumonia through the classification of genomic fingerprint pattern by pulsed-field gel electrophoresis of both oxacillin-resistant and oxacillin-susceptible Staphylococcus aureus isolates obtained from hospitalized patients in an intensive care unit.

Sensibilidade de isolados de candida spp. a antifúngicos por disco-difusão em ágar e microdiluição em caldo / Antifungal susceptibility of isolates of candida spp. by disk diffusion and broth microdilution

Os testes de sensibilidade aos antifúngicos realizados pelo método de disco-difusão em ágar são práticos e bem conhecidos pelos profissionais do laboratório de microbiologia, entretanto apresentam particularidades que os diferem dos testes realizados para bactérias. O objetivo deste trabalho foi comparar as técnicas de disco-difusão em ágar e microdiluição em caldo na determinação da sensibilidade in vitro de isolados de Candida spp. a antifúngicos. Foram analisados 63 isolados clínicos de leveduras, que incluíram as espécies Candida parapsilosis complex (n = 20), Candida albicans (n = 18), Candida tropicalis (n = 14), Candida glabrata (n = 4), Candida krusei (n = 4), Candida kefyr (n = 2) e Candida lusitaniae (n = 1). As técnicas de disco-difusão em ágar e de microdiluição em caldo foram utilizadas para testar a sensibilidade em relação aos antifúngicos fluconazol, itraconazol e anfotericina B. A sensibilidade ao voriconazol foi determinada somente pela técnica de disco-difusão. Os halos ao redor dos discos de fluconazol variaram de 14 mm a 50 mm, e a CIM de 0,125 µg/mL a 32 µg/mL; para itraconazol, os halos variaram de 9 mm a 27 mm e a CIM de 0,03 µg/mL a 0,25 µg/mL; para anfotericina B, 9 mm a 21mm e 0,5 µg/mL a 2 µg/mL, respectivamente; para voriconazol, o diâmetro dos halos variaram de 19 mm a 50 mm. Para as três espécies, C. albicans, C. parapsilosis e C. tropicalis, a técnica de disco-difusão apresentou boa concordância com a microdiluição, especialmente em relação ao fluconazol, representando, assim, um recurso importante para os laboratórios reportarem os resultados dos testes de sensibilidade dos isolados dessas espécies ao fluconazol.

Antimicrobial susceptibility tests performed by disk diffusion method are practical and well known by professionals that work in the microbiology laboratory. The disk diffusion methodology used to verify the susceptibility of fungi to antifungal agents, however, has characteristics that differ from the tests for bacteria. The objective of this study was to evaluate the disk diffusion method to determine the in vitro susceptibility to antifungal agents of Candida species. We analyzed 63 clinical isolates of yeasts, which included Candida parapsilosis complex species (n = 20), Candida albicans (n = 18), Candida tropicalis (n = 14), Candida glabrata (n = 4), Candida krusei (n = 4), Candida kefyr (n = 2) and Candida lusitaniae (n = 1). The susceptibility tests to antifungal drugs was performed by disk diffusion methods and broth microdilution for antifungal fluconazole, itraconazole and amphotericin B. Voriconazole was used to test the susceptibility only by the disk diffusion method. The inhibition halos of growth around disks of fluconazole ranged from 14 mm to 50 mm and the MIC from 0.125 µg/mL to 32 µg/mL, for itraconazole, halos ranged from 9 mm to 27 mm and the MIC from 0.03 µg/mL to 0.25 µg/mL, for amphotericin B, 9 mm to 21 mm and 0.5 µg/mL to 2 µg/mL, respectively. The diameter of voriconazole disks varied from 19 mm to 50 mm. For the three species, C. albicans, C. parapsilosis and C. tropicalis, the disk diffusion method showed good agreement with the microdilution, especially to fluconazole, thus representing an important resource for medical laboratories reporting results of susceptibility testing of isolates of these species to fluconazole.

Comparison of primers for RAPD-PCR from environmental isolates of Cryptococcus neoformans, Cryptococcus albidus and Cryptococcus laurentii complex

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.

Dermatophyte susceptibilities to antifungal azole agents tested in vitro by broth macro and microdilution methods / Suscetibilidade in vitro de dermatófitos a azóis pelos métodos macro e microdiluição em caldo

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 percent for itraconazole, 45 percent for fluconazole and 85 percent 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.

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 por cento para itraconazol, 45 por cento para fluconazol e 85 por cento 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.

Avaliação da produção de melanina por espécies de Cryptococcus em quatro diferentes meios de cultura / Evaluation of melanin production by Cryptococcus species in four different culture media

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 por cento 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 por cento produziram o pigmento em ágar batata e cenoura e L-dopa, 11,8 por cento em ágar alpiste, e 36 por cento em ágar girassol. De Cryptococcus laurentii, 53,8 por cento produziram em batata e cenoura e em semente de girassol, 61,5 por cento em L-dopa, 84,6 por cento 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 percent 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 percent produced the pigment in potato-carrot agar and L-dopa agar, 11.8 percent in Niger seed agar and 36 percent in sunflower seed agar. From Cryptococcus laurentii, 53.8 percent produced the pigment in potato-carrot agar and sunflower seed agar, 61.5 percent in L-dopa agar and 84.6 percent in Niger seed agar. Only one strain of Cryptococcus uniguttulatus presented slight production of the pigment, in potato-carrot agar.

Atividade enzimática, produção de slime e sensibilidade a antifúngicos de Candida sp / Enzymatic activity, slime production and antifungal agent sensitivity of Candida sp

A habilidade de Candida spp secretar enzimas extracelulares e slime tem sido associada como fatores de patogenicidade. Do total de 37 cepas de Candida sp, 100 por cento foram produtoras de proteinase, 83,8 por cento fosfolipase, 64,9 por cento slime e 100 por cento sensíveis ao fluconazol e itraconazol. Foram encontradas 17 tipagens (enzima/slime). Esta metodologia apresentou um bom índice discriminatório (D=0,93) podendo ser utilizado na caracterização fenotípica das leveduras.

Abilith of Candida spp to secrete extracellular enzymes and slime has been associated as pathogenicity factors. Out of a total of 37 strains of Candida sp, 100 percent were proteinase producers, 83.8 percent were phospholipase producers, 64.9 percent were slime producers and 100 percent were sensitive to fluconazole and itraconazole. Seventeen typings (enzymes/slime) were found. This methodology presented a good discrimination rate (D = 0.93) and could be used for phenotypic characterization of yeasts.

Spread of a methicillin-resistant staphylococcus aureus clone isolated from bacteremia in elderly patients in a Brazilian general hospital

A study of the genomic diversity of MRSA strains isolated from elderly patients with infection/colonization in three repeated prevalence, cross sectional studies was performed in the 1999-2000 period. In this study, 13 MRSA isolates from blood cultures and 5 from rectal and nare cultures were obtained from 18 patients (13 elderly and 5 adults). Most of the patients were being treated with two or more antimicrobials (83.3%), had insertion of invasive devices (88.9%) and were managed in ICU (Intensive Care Unit) and/or surgical units (66.7%). MIC (Minimum Inhibitory Concentration) data showed that 88.9% of the MRSA strains were resistant to high concentrations of oxacillin (MIC > 256 ìg/mL), 94.5% of the MRSA carried the mecA gene in their genome, and most (65.0%) of the isolates were indistinguishable according to their DNA finger­printing generated by PFGE (Pulsed-field gel electrophoresis). Although PFGE typing was performed with a few MRSA isolates, our results demonstrate that one MRSA clone was associated with infection/colonization in patients with an obvious connection among five out of eleven patients who stayed in the same clinic and ICU during the same period. Hospital acquired infection, a major “silent epidemy”, is associated with prolonged hospital stay and high mortality rate and its cause must be better evaluated.

Identification and molecular characterization of Van A-type vancomycin-resistant Enterococcus faecalis in Northeast of Brazil

The isolation of vancomycin resistant enterococci (VRE) in Brazil has rapidly increased, following the world wide tendency. We report in the present study the first isolation of vancomycin resistant Enterococcus faecalis (VRE) in the Northeast of Brazil. The four VRE isolates were characterized for antimicrobial susceptibility, genotypic typing by macro restriction of chromosomal DNA followed by pulsed-field gel electrophoresis and for characterization of the Tn1546-like element and plasmid contents. The isolates showed resistance to multiple antibiotics and a single genotype profile, suggesting the dissemination of a single clone among the patients. Tn1546 associated to genetic elements as plasmids shows the importance of infection control measures to avoid the spreading of glycopetide resistance by conjugative transfer of VanA elements.

Ocorrência de dermatófitos em amostras de unhas, pés e mãos coletadas de estudantes universitários / Occurrence of dermatophyte, in nails, feet and hands of universitary students

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 por cento) apresentaram positividade apenas pelo exame direto, e 20 (7,1 por cento) tiveram, além do exame direto positivo, crescimento de dermatófito, mediante cultivo da amostra biológica. T. rubrum foi o dermatófito isolado com maior freqüência (80 por cento), seguido por T. mentagrophytes (20 por cento). Considerando os sítios analisados neste trabalho, a ocorrência de dermatófitos foi observada em 10,4 por cento nas unhas dos pés, 5 por cento nas escamas de pés, 2,5 por cento nas unhas das mãos e apenas 0,4 por cento nas escamas das mãos.

In vitro susceptibility to antifungal agents of environmental Cryptococcus spp. isolated in the city of Ribeirão Preto, São Paulo, Brazil

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 æg/ml for amphotericin B, 0.06 and 8 æg/ml for itraconazole, and 0.5 and more than 64 æg/ml for fluconazole and 5-fluorocytosine. Agreement percentages for the two methodologies were 100 percent for amphotericin B and fluconazole for all the strains tested. For itraconazole, the agreement percentage was 81.3 percent in the C. neoformans strain and 100 percent for all the others. All species had a agreement percentage of 94.1 to 100 percent 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.