Environmental isolation of black yeast-like fungi involved in human infection

The present study focuses on potential agents of chromoblastomycosis and other endemic diseases in the state of Paraná, Southern Brazil. Using a highly selective protocol for chaetothyrialean black yeasts and relatives, environmental samples from the living area of symptomatic patients were analysed. Additional strains were isolated from creosote-treated wood and hydrocarbon-polluted environments, as such polluted sites have been supposed to enhance black yeast prevalence. Isolates showed morphologies compatible with the traditional etiological agents of chromoblastomycosis, e.g. Fonsecaea pedrosoi and Phialophora verrucosa, and of agents of subcutaneous or systemic infections like Cladophialophora bantiana and Exophiala jeanselmei. Some agents of mild disease were indeed encountered. However, molecular analysis proved that most environmental strains differed from known etiologic agents of pronounced disease syndromes: they belonged to the same order, but mostly were undescribed species. Agents of chromoblastomycosis and systemic disease thus far are prevalent on the human host. The hydrocarbon-polluted environments yielded yet another spectrum of chaetothyrialean fungi. These observations are of great relevance because they allow us to distinguish between categories of opportunists, indicating possible differences in pathogenicity and virulence.

In Vitro Susceptibility of Environmental Isolates of Exophiala dermatitidis to Five Antifungal Drugs

Several dematiaceous fungi frequently isolated from nature are involved in cases of superficial lesions to lethal cerebral infections. Antifungal susceptibility data on environmental and clinical isolates are still sparse despite the advances in testing methods. The objective of this study was to examine the activities of 5-flucytosine, amphotericin B, itraconazole, voriconazole and terbinafine against environmental isolates of Exophiala strains by minimum inhibition concentration (MIC) determination. The strains were obtained from hydrocarbon-contaminated soil, ant cuticle and fungal pellets from the infrabuccal pocket of attine gynes. Broth microdilution assay using M38-A2 reference methodology for the five antifungal drugs and DNA sequencing for fungal identification were applied. Terbinafine was the most active drug against the tested strains. It was observed that amphotericin B was less effective, notably against Exophiala spinifera, also studied. High MICs of 5-flucytosine against Exophiala dermatitidis occurred. This finding highlights the relevance of studies on the antifungal resistance of these potential opportunistic species. Our results also contribute to a future improvement of the standard methods to access the drug efficacy currently applied to black fungi. © 2012 Springer Science+Business Media Dordrecht.

Fungal diversity associated with Brazilian energy transmission towers

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

Selective factors involved in oil flotation isolation of black yeasts from the environment

The oil flotation isolation technique has been successfully applied to recover chaetothyrialean black yeasts and relatives from the environment. The selective mechanisms playing a role in isolation are unknown. The fungi concerned are supposed to occupy specialized microniches in nature, taking advantage of (1) oligotrophism. Mineral oil as a main selective agent may be based on (2) hydrophobicity or on (3) assimilation. All three hypotheses are tested in this paper. Results show that cell wall hydrophobicity is unlikely to be a selective factor. Incubation under poor nutrient conditions provides competitive advantage for black yeasts, especially for Exophiala strains, which are subsequently enriched by mineral oil which enhances growth in this group of fungi. Incubation under mineral media and mineral oil can be used as selective factor.

Isolation and identification of black yeasts by enrichment on atmospheres of monoaromatic hydrocarbons

Black yeast members of the Herpotrichiellaceae present a complex ecological behavior: They are often isolated from rather extreme environments polluted with aromatic hydrocarbons, while they are also regularly involved in human opportunistic infections. A selective technique to promote the in vitro growth of herpotrichiellaceous fungi was applied to investigate their ecophysiology. Samples from natural ecological niches and man-made environments that might contain black yeasts were enriched on an inert solid support at low humidity and under a controlled atmosphere rich in volatile aromatic hydrocarbons. Benzene, toluene, and xylene were provided separately as the sole carbon and energy source via the gas phase. The assayed isolation protocol was highly specific toward mesophilic Exophiala species (70 strains of this genus out of 71 isolates). Those were obtained predominantly from creosote-treated railway ties (53 strains), but isolates were also found on wild berries (11 strains) and in guano-rich soil samples (six strains). Most of the isolates were obtained on toluene (43 strains), but enrichments on xylene and benzene also yielded herpotrichiellaceous fungi (17 and 10 isolates, respectively). Based upon morphological characterizations and DNA sequences of the full internal transcriber spacers (ITS) and the 8.5S rRNA genes, the majority of the obtained isolates were affiliated to the recently described species Exophiala xenobiotica (32 strains) and Exophiala bergeri (nine strains). Members of two other phylogenetic groups (24 and two strains, respectively) somewhat related to E. bergeri were also found, and a last group (three strains) corresponded to an undescribed Exophiala species. © 2010 The Author(s).

Modelagem matemática da doença do caranguejo letárgico via ondas viajantes

Pós-graduação em Biometria - IBB

Influência das condições de cultivo e métodos de extração na produção de metabólitos antioxidantes por fungos isolados do litoral paulista

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Avaliação do método de Iwatsu et al., (1981) para isolamento de leveduras negras do solo, degradadoras de hidrocarbonetos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Fungos negros derivados de Atta spp: diversidade e fatores de virulência

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

Leaf-cutting ants: an unexpected microenvironment holding human opportunistic black fungi

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

Micro-fungos de interesse para o setor de petróleo, gás e biocombustíveis

Oil is a hydrocarbon mixture of various sizes, including saturated and aromatic compounds. Natural gas is a mixture of gaseous hydrocarbons and its main component is methane. In our society, the great demand for these fuels requires fast extraction, transportation and refining, increasing the number of accidents that compromise the environment. Oil is a finite resource and it is necessary to reduce the problems related to the question concerning environmental pollution which has encouraged the search for alternative fuel sources in our country. So today we have two major biofuels: ethanol and biodiesel. Concurrently, many studies have been done directed toward the isolation of microorganisms capable of degrading petrochemical industrial wastes, most of them using as a source of isolation soil and water collected in a contaminated environment. Isolation from alternative substrates has emerged as a new strategy that has provided satisfactory results. In this work, we present the leaf-cutter ants of the Attini tribe as a source for the isolation of micro-fungi with the potential for hydrocarbon degradation. These insects have a social way of life and a highly specialized system of intra and interspecific communication, which is based on the recognition of individuals through volatile chemical compounds, the majority hydrocarbons, stored in their exoskeleton. The micro-environment exoskeleton of Attini ants (genus Atta) used in this work proved to be a rich source of microbial biodiversity, as other studies have found. The flotation isolation technique applied here allowed the achievement of 214 micro-fungi, 118 representatives of the dematiaceous fungi group and 96 hyaline filamentous fungi. They were submitted to toluene degradation tests and at least one strain of each genus presented good results, namely Teratosphaeria, Exophiala, Cladosporium, Penicillium, Aspergillus... (Complete abstract click electronic access below)

Fungos fitopatógenos associados às formigas cortadeiras (Formicidae: Tribo Attini)

The ants of genus Atta belong to the Attine tribe (order Hymenoptera, family Formicidae, subfamily Myrmicinae) and are commonly known as leaf-cutting ants for having the habit of cutting several vegetable species used as substrate for growing mutualistic fungus (Agaricales: Lepiotaceae). Recent studies showed that, in addition to that, other fungi may occur in the nests in a dorment state or participate in the functioning dynamic of this symbiosis. Researches related to surveys of fungus biodiversity in nests of different Atta species have found important phytopathogenic representatives. In Brazil, studies about integrated management of plagues, developed by Embrapa Meio Ambiente (Embrapa Environment), point out the need of higher investments in projects that involve the phytopathogenic transmission by insects in order to reduce costs to control them or minimize environmental impact. The purpose of this study was to broaden the knowledge about the ecology of these fungi, isolating and identifying species associated with Attine tribe ants, thus understanding the scope of pathogenic and phytopathogenic species spread by these ants. For that reason, gynes were collected from Atta laevigata and Atta capiguara anthills located at Unesp Botucatu (São Paulo, Brazil) campus. In order to isolate the fungus, the mineral oil floating technique was used. The identification of the isolated fungi was done based on microscopic and molecular characteristics using DNA ribosomal sequencing. The most highly abundant genera found so far were: Cladosporium, Exophiala, Penicillium, Acremonium, Phialophora and Teratosphaeria. Representatives of the genera Exophiala, Phialophora and Cladosporium may be human pathogens, whereas Teratosphaeria and Penicillium are related to diseases in Eucalyptus and citric fruits, respectively. The results show that these ants may host important fungal species besides the ones already... (Complete abstract click electronic access below)

Ocorrência de fungos negros em ambientes domésticos

Black fungi are able to adapt to extreme environmental conditions, such as: high temperatures, the presence of toxic chemical substances and lack of nutrients. Besides, they are also potential pathogens to humans. The natural environment of many black fungi is still unknown and some studies are being conducted to evaluate the biodiversity of this group and their different habitats. This study aimed to isolate black fungi in domestic environments and facilities, such as toothbrushes, fridge sealing rubbers, bathroom strainers and divisions, windows, wall tiles and bath sponge. For the collection, material surfaces were scratched with a scalpel and the resulting fragments were sewed in Mycosel agar (DifcoTM), supplemented with actidione to inhibit the growth of highly-sporulating fungi. Plates were incubated at 25ºC for three weeks. The 46 isolated fungi were maintained on MA2% slants at 8ºC and cryopreserved at -80ºC. Fungal identification was performed through the analysis of macro and microscopic features and ITS rDNA sequencing. The following black fungi taxa were found: Ascomycota sp., Cladosporium spp., Dothideomycete sp., Exophiala alcalophila, Ochroconis mirabilis and Rhinocladiella atrovirens. Non-melanized fungi were also found, such as Geosmithia sp., Penicillium sp. and Rhodotorula mucilaginosa. The temperature tests showed that isolated black fungi were not able to grow at 37°C, however, this temperature proved to be fungistatic to 43% of them. According to literature, all black fungi isolated in this study are opportunistic pathogens and additional studies are necessary to evaluate the risk that these micro-organisms offer to health, once they were isolated from domestic environments