A new culture medium for recovering the agents of Cryptococcosis from environmental sources.

The isolation of Cryptococcosis agents from environmental samples may be difficult due to the presence of groups of fast-growing fungi. We propose a new culture medium based on a modification of Dichloran Rose-Bengal Chloramphenicol Medium (DRBCm) to detect colonies of Cryptococcus neoformans. Our results indicate that DRBCm is superior to the classical Bird Seed Agar in its ability to detect colonies of C. neoformans.

A new culture medium for recovering the agents of Cryptococcosis from environmental sources

The isolation of Cryptococcosis agents from environmental samples may be difficult due to the presence of groups of fast-growing fungi. We propose a new culture medium based on a modification of Dichloran Rose-Bengal Chloramphenicol Medium (DRBCm) to detect colonies of Cryptococcus neoformans. Our results indicate that DRBCm is superior to the classical Bird Seed Agar in its ability to detect colonies of C. neoformans.

Diferentes intervalos de tempo de leitura do sistema API 20C AUX® na identificação de leveduras de interesse médico / Different reading time intervals of the API 20C AUX® system for identifying the yeasts of medical interest

Infecções de corrente sanguínea por leveduras do gênero Candida são uma das principais causas de morbidade e mortalidade em pacientes imunocomprometidos. Candida albicans permanece a espécie mais isolada nestas infecções e é de fácil e rápida identificação. Contudo, existem outras espécies, como C. parapsilosis, C. tropicalis, C. glabrata e C. krusei, que são encontradas com menor frequência e que necessitam de maior período de tempo e de metodologias comerciais automatizadas ou semi-automatizadas para sua identificação. Neste estudo foram analisadas 146 cepas de leveduras quanto à capacidade do Sistema API 20C AUX® (Biomerieux®, França) em identificar corretamente o gênero e a espécie de microrganismos em diferentes períodos de leitura, visando-se a liberação do resultado em menor tempo. C. parapsilosis, C. guilliermondii, C. pelliculosa, C. colliculosa, Rhodotorula mucilaginosa, Saccharomyces cerevisae, Trichosporon mucoides e T. asahii foram as leveduras cujos resultados finais puderam ser liberados nos períodos de tempo de 96, 120 e 144 h. Oitenta por cento das C. glabrata e 69 % das C. tropicalis também foram identificadas nos períodos além do tempo estabelecido. Com os resultados obtidos é possível antecipar a identificação do gênero e de algumas espécies de leveduras...

Comparison of the broth microdilution (BMD) method of the European Committee on Antimicrobial Susceptibility Testing and the Clinical Laboratory Standards Institute BMD method for non-Candida albicans and non-C. tropicalis bloodstream isolates from eleven tertiary hospitals in São Paulo state, Brazil.

We aim in this study to provide levels of susceptibility of 162 bloodstream isolates of non-Candida albicans and non-C. tropicalis species from a sentinel program conducted in 11 hospitals in Brazil. Additionally, we compared the broth microdilution (BMD) method of the European Committee of Susceptibility Testing (EUCAST) with Clinical Laboratory Standards Institute (CLSI) BMD method for fluconazole, itraconazole, voriconazole, and amphotericin B. The study included 103 C. parapsilosis, 38 C. glabrata, 8 C. orthopsilosis, and 7 C. krusei isolates, and single isolates of Pichia anomala, C. famata, C. lusitaniae, C. kefyr, C. guilliermondii, and C. metapsilosis. Of note, we observed cross-resistance between fluconazole and voriconazole for two isolates being one C. parapsilosis and one C. glabrata. Good essential agreement (EA) was observed between the EUCAST and the CLSI results for C. parapsilosis and for fluconazole, itraconazole, voriconazole, and amphotericin B, respectively: 98%, 99%, 98%, and 97%. Otherwise, for C. glabrata, the EA for fluconazole was 84.2% and for voriconazole 89.4%. Because data from Brazil are scarce, our results contribute to the consolidation of the database of candidemia agents and monitoring of trends in the profile of drug resistance.

Indoor and outdoor atmospheric fungal spores in the São Paulo metropolitan area (Brazil): species and numeric concentrations.

The aim of this study was to estimate the indoor and outdoor concentrations of fungal spores in the Metropolitan Area of Sao Paulo (MASP), collected at different sites in winter/spring and summer seasons. The techniques adopted included cultivation (samples collected with impactors) and microscopic enumeration (samples collected with impingers). The overall results showed total concentrations of fungal spores as high as 36,000 per cubic meter, with a large proportion of non culturable spores (around 91% of the total). Penicillium sp. and Aspergillus sp. were the dominant species both indoors and outdoors, in all seasons tested, occurring in more than 30% of homes at very high concentrations of culturable airborne fungi [colony forming units(CFU) m(-3)]. There was no significant difference between indoor and outdoor concentrations. The total fungal spore concentration found in winter was 19% higher than that in summer. Heat and humidity were the main factors affecting fungal growth; however, a non-linear response to these factors was found. Thus, temperatures below 16 degrees C and above 25 degrees C caused a reduction in the concentration (CFU m(-3)) of airborne fungi, which fits with MASP climatalogy. The same pattern was observed for humidity, although not as clearly as with temperature given the usual high relative humidity (above 70%) in the study area. These results are relevant for public health interventions that aim to reduce respiratory morbidity among susceptible populations.

Cerebral aspergillosis due to Aspergillus fumigatus in AIDS patient: first culture-proven case reported in Brazil.

Cerebral aspergillosis is a rare cause of brain expansive lesion in AIDS patients. We report the first culture-proven case of brain abscess due to Aspergillus fumigatus in a Brazilian AIDS patient. The patient, a 26 year-old male with human immunodeficiency virus (HIV) infection and history of pulmonary tuberculosis and cerebral toxoplasmosis, had fever, cough, dyspnea, and two episodes of seizures. The brain computerized tomography (CT) showed a bi-parietal and parasagittal hypodense lesion with peripheral enhancement, and significant mass effect. There was started anti-Toxoplasma treatment. Three weeks later, the patient presented mental confusion, and a new brain CT evidenced increase in the lesion. He underwent brain biopsy, draining 10 mL of purulent material. The direct mycological examination revealed septated and hyaline hyphae. There was started amphotericin B deoxycholate. The culture of the material demonstrated presence of the Aspergillus fumigatus. The following two months, the patient was submitted to three surgeries, with insertion of drainage catheter and administration of amphotericin B intralesional. Three months after hospital admission, his neurological condition suffered discrete changes. However, he died due to intrahospital pneumonia. Brain abscess caused by Aspergillus fumigatus must be considered in the differential diagnosis of the brain expansive lesions in AIDS patients in Brazil.

Cerebral aspergillosis due to Aspergillus fumigatus in AIDS patient: first culture - proven case reported in Brazil

La aspergilosis cerebral es una causa rara de lesión expansiva cerebral en pacientes con SIDA. Presentamos el primer reporte de un absceso cerebral causado por Aspergillus fumigatus en un paciente brasileño con SIDA. El paciente, de 26 años de edad, presentaba antecedentes de infección por el virus de la inmunodeficiencia humana (VIH), tuberculosis pulmonar y toxoplasmosis cerebral. Manifestó fiebre, tos, disnea y dos episódios de convulsiones. La tomografía computadorizada (TC) demostró una lesión hipodensa parasagital y bi-parietal con realce periférico e importante efecto de masa. Se inició tratamiento anti-Toxoplasma. Tres semanas después, el paciente evidenció confusión mental y una nueva TC de cráneo mostró aumento de la lesión. Se realizó biopsia cerebral con drenaje de 10 mL de material purulento. El examen micológico directo reveló hifas hialinas septadas. Se inició anfotericina B deoxicolato. La cultura del material demostró presencia de Aspergillus fumigatus. En los siguientes dos meses el paciente fue sometido a otras tres cirugías, insertándose un catéter de drenaje y administrándose anfotericina B intralesional. Tres meses después de la admisión hospitalaria, la condición neurológica del paciente sufrió discretos cambios. Sin embargo, falleció debido a neumonia intrahospitalaria. Aunque muy raros, los abscesos cerebrales causados por Aspergillus fumigatus deben ser considerados en el diagnóstico diferencial de las lesiones expansivas cerebrales en pacientes con SIDA.

Extracellular enzymatic activity and serotype of Cryptococcus neoformans strains isolated from AIDS patients in Brazil.

One hundred and fifty-one Cryptococcus neoformans strains isolated from AIDS patients in Brazil and maintained in the Adolfo Lutz Institute (São Paulo, Brazil) were tested for phospholipase, protease and other extracellular enzymatic activities and their serotypes determined. Production of extracellular phospholipase and protease was tested by the agar plate methods. Determination of extracellular enzyme profile of the strains was performed by using the API-ZYM kit system, which can test 19 different enzymes. The serotypes were determined by cell agglutination using the Crypto-check method. Among the 151 strains, 147 were identified as serotype A and four strains were serotype AD. Production of extracellular phospholipase and protease was extensive and observable at early stages of incubation. All of the tested strains were positive for the production of both enzymes. In the API-ZYM tests, more than 90 % of the 151 tested strains were positive for esterase C4 (No. 3), esterase lipase C8 (No. 4), leucine arylamidase (No. 6), phosphatase acid (No. 11), naphtol-AS-BI-phosphohydrolase (No. 12), alpha-glucosidase (No. 16) and beta-glucosidase (No. 17). Differences in enzymatic activities between the Brazilian strains and strains isolated in other countries were observed. The phospholipase, protease and other enzyme activities may play a role in host tissue invasion by C. neoformans.

Infecçäo hospitalar por leveduras em unidade de terapia intensiva infantil: abordagem epidemiológica / Nosocominal infection by yeats in pediatric intensive care unit

A incidência de infecçöes fúngicas tem aumentado no mundo todo nos últimos anos. Estas infecçöes säo causas importantes de morbidade e mortalidade, particularmente, em pacientes de risco internados em unidades de terapia intensiva. Objetivos: (1) Estudar a distribuiçäo de espécies de leveduras, em estado de colonizaçäo e infecçäo, isoladas de pacientes de uma UTI infantil; (2) Estudar o perfil de sensibilidade destas espécies e (3) identificar os genótipos das leveduras. Material e Métodos: A leveduras foram identificadas com kit de API 20C aux (Biolab Merieux, Fr) e provas fisiológicas adicionais. PFGE foi usado para genotipagem e a sensibilidade foi verificada segundo M27A (NCCLS 1997) modificado por EUCAST. Resultados: A distribuiçäo das 82 amostras de leveduras obtidas de 34 pacientes foi: Candida Albicans (55 por cento), C.tropicalis e C.parapsilosis (15 por cento), C.Krusei (9 por cento), Trichosporon asahii (5 por cento) e C. glabrata (1 por cento). C. Albicans (45 por cento) e C.parapsilosis (36 por cento) foram as espécies mais envolvidas em infecçäo hospitalar (IH). As amostras foram sensíveis a anfoB (100 por cento), FLUCO (83,7 por cento) e ITRA (90 por cento), entretanto C.tropicalis apresentou alta resistência (46,7 por cento) aos antifúngicos azólicos. Em 6 pacientes a cariotipagem permitiu verificar a correspondência entre os genótipos dos agentes etiológicos de IH e aqueles das cepas colonizadoras. Conclusöes: Candida Albicans e C. parapsilosis, sensíveis aos antifúngicos testados, foram os agentes mais freqüentes em episódios de IH. A tipagem molecular permitiu associar os subtipos de cepas colonizadoras com os agentes etiológicos de infecçäo em 6 pacientes

Rhodotorula Rubra em cabelos humanos simulando quadro de piedra branca / Similar white piedra by Rodotorula rubra

Resumo: Rhodotorula rubra é levedura oportunista, esando associada a uma variedade de processos patológicos no homem, é frequêntemente encontrada como contaminante da pele, pulmäo, urina, fezes e sistema nervoso central e com grande significado quando presente no sangue. As determinantes humanas causadas por Rhodotorula têm sido relatadasmas näo documentadas. O presente trabalho relata pela primeira vez em nosso meio, o isolamento desse microrganismo, como agente etiológico de infecçäo de cabelos humanos que ao exame clínico, simula pidra branca. Cabelos apresentando nódulos claros foram colhidos de paciente com 18 anos de idade. Em seguida classificados com KOH a 20 (por cento) entre lâmina e lamínula e observados no microscópio óptico comum e na microscopia eletrônicade varredura. Os cabelos foram semeados em ágar Sabnouraud-glicose contendo 200 mg/l de clonranfenicol, incubados à 25 graus C por 5 dias. Após o isolamento da levedura, a mesma foi identificada utilizando-se técnicas micológicas usuais. O exame microscópico dos pêlos infectados revelou em algumas regiöes o aumento do volume do cabelo, característico da tricorrexe nodosa. Em outras regiöes os pêlos já se apresentavam fragmentados e, com relativa facilidade podia-se observar leveduras arredondadas com brotamento em base estreita. A microscopia eletrônica de varredura pode-se observar mais detalhadamente a descriçäo acima. A levedura isolada foi identificada como Rhodotorula rubra, sem atividade lipolítica. É importante ressaltar que a referida levedura foi isolada repetidas vezes em cultura pura dos cabelos infectados. Este achado clínicoe laboratorial deve seer considerado, assim como deve merecer a devida atençäo dos pesquisadoresos isolamentos de R. rubra os quais permitiräo um correto diagnóstico micológico e terapêutico adequada (au)