Molecular typing and antifungal susceptibility of clinical sequential isolates of Cryptococcus neoformans from São Paulo State, Brazil

The antifungal susceptibility profiles and the genetic variability of 83 sequential clinical isolates of Cryptococcus neoformans, including four Cryptococcus gattii isolates, obtained from 38 São Paulo AIDS patients with cryptococcal meningitis were assessed by electrophoretic karyotyping and random amplified polymorphic DNA (RAPD) analysis. The majority of the Cryptococcus neoformans isolates were highly susceptible to amphotericin B and fluconazole. Twenty percent of the minimum inhibitory concentration values for amphotericin B varied from 0.5 to 1 mu g mL(-1). For fluconazole, 22% occurred in the range 8-16 mu g mL(-1). Sequential isolates from nine patients showed a trend towards lower susceptibility to fluconazole, flucytosine, itraconazole and amphotericin B. The results of molecular typing by electrophoretic karyotyping and RAPD analysis showed the presence of 22 electrophoretic karyotypes (EK) and 15 RAPD profiles that were highly correlated. Our results provided evidence for the occurrence of genetic changes in some strains associated with microevolution during the course of infection. We also observed both microevolution and simultaneous coinfection with two distinct Cryptococcus neoformans strains in one patient. In some patients, we found changed EK- and RAPD patterns in association with increased MIC values.

Genotyping, serotyping and determination of mating-type of Cryptococcus neoformans clinical isolates from São Paulo State, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Composition and antifungal activity against Candida albicans, Candida parapsilosis, Candida krusei and Cryptococcus neoformans of essential oils from leaves of Piper and Peperomia species

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Molecular typing of IberoAmerican Cryptococcus neoformans isolates

A network was established to acquire basic knowledge of Cryptococcus neoformans in IberoAmerican countries. To this effect, 340 clinical, veterinary, and environmental isolates from Argentina, Brazil, Chile, Colombia, Mexico, Peru, Venezuela, Guatemala, and Spain were typed by using M13 polymerase chain reaction-fingerprinting and orotidine monophosphate pyrophosphorylase (URA5) gene restriction fragment length polymorphsm analysis with Hhal and Sau961 in a double digest. Both techniques grouped all isolates into eight previously established molecular types. The majority of the isolates, 68.2% (n=232), were VNI (var. grubii, serotype A), which accords with the fact that this variety causes most human cryptococcal infections worldwide. A smaller proportion, 5.6% (n=19), were VNII (var. grubii, serotype A); 4.1% (n=14), VNIII (AD hybrid), with 9 isolates having a polymorphism in the URA5 gene; 1.8% (n=6), VNIV (var. neoformans, serotype D); 3.5% (n=12), VGI; 6.2% (n=21), VGII; 9.1% (n=31), VGIII, and 1.5% (n=5) VGIV, with all four VG types containing var. gattii serotypes B and C isolates.

Sensibilidade à antifúngicos sintéticos e naturais anti-Cryptococcus neoformans determinados por citometria de fluxo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Perfil fenotípico e de expressão de proteínas de Cryptococcus neoformans após tratamento com substâncias obtidas da planta Pterogyne nitens

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Caracterização do gene ERG11 envolvido na resistência de Cryptococcus neoformans e Cryptococcus gatti as drogas azólicas e identificação de novos alvos a partir de produtos naturais

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

First isolation of Cryptococcus gattii molecular type VGII and Cryptococcus neoformans molecular type VNI from environmental sources in the city of Belém, Pará, Brazil

Cryptococcus neoformans and Cryptococcus gattii are important agents of meningoencephalitis in humans in the city of Belém. This clinical data suggests that the region may be a highly endemic area for the pathogenic Cryptococcus species within the state of Pará (PA), Northern Brazil. Preliminary analysis of 11 environmental samples from the city of Belém showed two positive locations, including a hollow of a kassod tree (Senna siamea) colonized simultaneously by C. gattii molecular type VGII and C. neoformans molecular type VNI, and a birdcage in a commercial aviary positive for C. neoformans, molecular type VNI. This is the first evidence of an environmental occurrence of molecular types VNI and VGII in PA.

Neurocriptococose pediátrica no Estado do Pará: espectro de achados tomográficos na infecção por Cryptococcus neoformans var. gattii

Este estudo mostra o espectro de lesões cerebrais, através de tomografia computadorizada, na neurocriptococose da infância, por Cryptococcus neoformans var. gattii, no Estado do Pará. Analisamos os achados tomográficos de onze crianças (menores de 13 anos de idade), com infecção comprovada do sistema nervoso central por Cryptococcus neoformans var. gattii, internados no Hospital Universitário João de Barros Barreto, Belém PA, entre janeiro de 1992 a dezembro de 2000. A neurocriptococose foi definida pela identificação de leveduras encapsuladas ao exame microscópico, isolamento de Cryptococcus neoformans do líquor e identificação positiva da variedade gattii, através do meio composto de canavanina, glicina e azul-de-bromotimol com, pelo menos, o estudo tomográfico no momento do diagnóstico. A idade das crianças estudadas variou entre 6 a 12 anos, com média de 8,8 anos. Cincos eram meninos e seis meninas. Os principais achados clínicos foram cefaléia, febre e rigidez de nuca (n=11), náuseas e vômitos (n=10). O tempo médio entre o início dos sintomas e o diagnóstico foi de 4,2 semanas (variando de 2 a 8 semanas). Todas as tomografias de crânio foram anormais. Em todos os pacientes foram observados nódulos hipodensos. As demais anormalidades tomográficas observadas foram: 6 pacientes com hidrocefalia, 9 com atrofia difusa e 5 com associação de hidrocefalia e atrofia difusa. Descreve-se, pela primeira vez, achados tomográficos em série de casos de neurocriptococose por Cryptococcus neoformans var. gattii em crianças imunocompetentes. Este estudo mostra que esta infecção desenvolve múltiplos nódulos hipodensos, especialmente na região dos gânglios da base e na substância branca cerebral. As lesões, aparentemente progridem para importante atrofia da substância branca, com dilatação ventricular e proeminência de sulcos cerebrais, provavelmente, conseqüência de hidrocefalia compensatória. Em geral, estes pacientes apresentaram alterações leves do córtex cerebral.

Cryptococcus neoformans de fontes ambientais e biopropecção de extratos de plantas com atividade saneante

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Tipagem molecular, perfis de sensibilidade e caracterização de transcritos diferencialmente expressos durante a infecção de Cryptococcus neoformans

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Composition and antifungal activity against Candida albicans, C. parapsilopisis, Candida krusei and Cryptococcus neoformans of essential oils from leaves of Piper and Peperomia species

This study was addressed to investigate the composition and antifungal activity of essential oils from leaves of Piperaceae species (Piper aduncum, Piper amalago, Piper cernuum, Piper diospyrifolium, Piper crassinervium, Piper gaudichaudianum, Piper solmsianum, Piper regnellii, Piper tuberculatum, Piper umbelata and Peperomia obtusifolia) against Candida albicans, C. parapsilosis, C. krusei and Cryptococcus neoformans. The essential oils from P. aduncum, P. gaudichaudianum and P. solmsianum showed the highest antifungal activity against Cryptococcus neoformans with the MIC of 62.5, 62.5 and 3.9 mg.mL-1, respectively. The oil from P. gaudichaudianum showed activity against C. krusei with MIC of 31.25 mg.mL-1.

Estudo dos mecanismos de resistência e virulência de isolados de Cryptococcus neoformans e Cryptococcus gattii e quantificação de genes de bomba de efluxo pós tratamento com Galatos de Alquila

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

The homeostasis of iron, copper, and zinc in paracoccidioides brasiliensis, cryptococcus neoformans var. Grubii, and cryptococcus gattii: a comparative analysis

Iron, copper, and zinc are essential for all living organisms. Moreover, the homeostasis of these metals is vital to microorganisms during pathogenic interactions with a host. Most pathogens have developed specific mechanisms for the uptake of micronutrients from their hosts in order to counteract the low availability of essential ions in infected tissues. We report here an analysis of genes potentially involved in iron, copper, and zinc uptake and homeostasis in the fungal pathogens Paracoccidioides brasiliensis, Cryptococcus neoformans var. grubii, and Cryptococcus gattii. Although prior studies have identified certain aspects of metal regulation in Cryptococcus species, little is known regarding the regulation of these elements in P. brasiliensis. We also present amino acid sequences analyses of deduced proteins in order to examine possible conserved domains. The genomic data reveals, for the first time, genes associated to iron, copper, and zinc assimilation and homeostasis in P. brasiliensis. Furthermore, analyses of the three fungal species identified homologs to genes associated with high-affinity uptake systems, vacuolar and mitochondrial iron storage, copper uptake and reduction, and zinc assimilation. However, homologs to genes involved in siderophore production were only found in P. brasiliensis. Interestingly, in silico analysis of the genomes of P. brasiliensis Pb01, Pb03, and Pb18 revealed significant differences in the presence and/or number of genes involved in metal homeostasis, such as in genes related to iron reduction and oxidation. The broad analyses of the genomes of P. brasiliensis, C. neoformans var. grubii, and C. gattii for genes involved in metal homeostasis provide important groundwork for numerous interesting future areas of investigation that are required in order to validate and explore the function of the identified genes and gene pathways.