Black yeasts from the slope sediments of Bay of Bengal: phylogenetic and functional characterization

Occurrence of black yeasts in the slope sediments of Bay of Bengal was investigated during FORV Sagar Sampada cruises 236 and 245. The black yeast population was found to be very scanty in the area and the isolates could be obtained from 200m to 1000m depth regions in the slope sediments. The isolates were identified as Hortaea werneckii by Internal Transcribed Spacer (ITS) sequencing. The biodegradation potential of these strains was found to be very high with all the strains exhibiting protease, lipase and amylase production. The optimum growth conditions were pH 8, salinity 30 ppt and temperature 30oC. The pigment melanin, in these organisms was identified to be of dihydroxynaphthalene type by NMR. The melanin was found to exhibit inhibitory activity against different human and fish pathogens. Melanin degrading enzyme could also be extracted from these organisms

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).

Marine Yeasts from the Slope Sediments of Arabian Sea and Bay of Bengal

The present study provides an account of the occurrence and diversity of marine yeasts in the slope sediments of Arabian Sea and Bay of Bengal. It also gives a clear idea about the role of yeasts in the benthic realm of marine ecosystem. The lipolytic potential of the organisms indicate the presence of rich lipid moieties in the study area. The isolates, Candida sp. SD 302 and Pichia guilliermondii SD 337 were proved to have potential oil degrading property and can be employed as bioremediators of oil spill after further characterization. The black yeasts isolated during the study area were found to have high commercial value by virtue of the by-products obtained from them. The melanin and the melanin degrading enzyme extracted from these organisms are potential bioactive materials for application in cosmetology.

Yeasts associated with nests of the leaf-cutting ant Atta sexdens rubropilosa Forel, 1908

A total of 137 yeasts associated with the leaf-cutting ant Atta sexdens rubropilosa Forel, 1908 were characterized, being selected 93 for analysis. Twenty four species belonging to seven genera(Candida, Cryptococcus, Rhodotorula, Sporobolomyces, Tremella, Trichosporon, Pichia) were isolated from the different analysed material. The genus Candida was widely distributed, with C. homilentoma, C. colliculosa-like, C. famata and C. colliculosa being the most prevalent. A few isolates did not fit the standard descriptions and probably some of them could be new biotypes or even new species. Three strains of black yeasts were also isolated, and four others were identified as being Candida spp. The effective number of yeast species was higher in newer sponge. The origin, distribution and relative importance of these microorganisms for the ants are discussed.

Ascomycetous Yeasts Associated with Naturally Occurring Fruits in a Tropical Rain Forest

Fruits from twenty different species of angiosperms were collected during the period from November, 1991 to January, 1992. Two hundred and two strains of yeasts and yeast-like fungi were isolated, of which 74 % showed ascomycetic affinity. Candida was the predominant genus, followed by (in descending order of occurrence): Cryptococcus, Klœckera, Sporobolomyces, Pichia, Hanseniaspora and Bullera. Black yeasts and other strains showing basidiomycetic affinity were also isolated. The genus Candida represented the highest number of identified species and the greatest variety of associated substrates. Among the ascomycetes and their anamorphs, 38 species were identified, with Klœckera apiculata being the most frequent among the isolates and the one which occurred in the largest variety of substrates. Some of the biotypes designated as Candida sp. A, B, C, D, E, F, G, H, I, and Pichia sp. did not correspond to the standard species description found in the literature, and may represent new species. The strains of yeasts isolated in this study were characterized and incorporated into the Tropical Culture Collection of the Fundação Tropical de Pesquisas e Tecnologia André Tosello, Campinas, São Paulo.

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.

Cladophialophora saturnica sp nov., a new opportunistic species of Chaetothyriales revealed using molecular data

While many members of the black yeasts genus Cladophialophora have been reported to cause diseases in humans, understanding of their natural niche is frequently lacking. Some species can be recovered from the natural environment by means of selective isolation techniques. The present study focuses on a Cladophialophora strain that caused an interdigital tinea nigra-like lesion in a HIV-positive Brazilian child. The fungal infection was successfully treated with oxiconazole. Similar strains had been recovered from the environment in Brazil, Uruguay and the Netherlands. The strains were characterized by sequencing the Internal Transcribed Spacer (ITS) regions and the small subunit (SSU) of the nuclear ribosomal RNA gene, as well as the elongation factor 1-alpha (EF1) gene. Since no match with any known species was found, it is described as the new species, Cladophialophora saturnica.

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

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

Novel Phialophora species from leaf-cutting ants (tribe Attini)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Statnote 36: Do the data fit the Poisson distribution

In previous Statnotes, many of the statistical tests described rely on the assumption that the data are a random sample from a normal or Gaussian distribution. These include most of the tests in common usage such as the ‘t’ test ), the various types of analysis of variance (ANOVA), and Pearson’s correlation coefficient (‘r’) . In microbiology research, however, not all variables can be assumed to follow a normal distribution. Yeast populations, for example, are a notable feature of freshwater habitats, representatives of over 100 genera having been recorded . Most common are the ‘red yeasts’ such as Rhodotorula, Rhodosporidium, and Sporobolomyces and ‘black yeasts’ such as Aurobasidium pelculans, together with species of Candida. Despite the abundance of genera and species, the overall density of an individual species in freshwater is likely to be low and hence, samples taken from such a population will contain very low numbers of cells. A rare organism living in an aquatic environment may be distributed more or less at random in a volume of water and therefore, samples taken from such an environment may result in counts which are more likely to be distributed according to the Poisson than the normal distribution. The Poisson distribution was named after the French mathematician Siméon Poisson (1781-1840) and has many applications in biology, especially in describing rare or randomly distributed events, e.g., the number of mutations in a given sequence of DNA after exposure to a fixed amount of radiation or the number of cells infected by a virus given a fixed level of exposure. This Statnote describes how to fit the Poisson distribution to counts of yeast cells in samples taken from a freshwater lake.

Development of a novel PCR assay for the identification of the black yeast, Exophiala (Wangiella) dermatitidis from adult patients with cystic fibrosis (CF)

Cystic fibrosis (CF) patients may suffer increased morbidity and mortality through colonisation, allergy and invasive infection from fungi. The black yeast, Exophiala dermatitidis (synonym Wangiella dermatitidis) has been found with increasing frequency in sputum specimens of CF patients, with reported isolation rates ranging from 1.1 to 15.7%. At present, no diagnostic PCR exists to aid with the clinical laboratory detection and identification of this organism. A novel species-specific PCR-based assay was developed for the detection of E. dermatitidis, based on employment of rDNA operons and interspacer (ITS) regions between these rDNA operons. Two novel primers, (designated ExdF & ExdR) were designed in silico with the aid of computer-aided alignment software and with the alignment of multiple species of Exophiala, as well as with other commonly described yeasts and filamentous fungi within CF sputum, including Candida. Aspergillus and Scedosporium. An amplicon of approximately 455 by was generated, spanning the partial ITS I region - the complete 5.8S rDNA region - partial ITS2 region, employing ExdF (forward primer [16-mer], 5'-CCG CCT ATT CAG GTC C-3' and ExdR (reverse primer [16-mer], 5'-TCT CTC CCA CTC CCG C-3', was employed and optimised on extracted genomic DNA from a well characterised culture of E. dermatitidis, as well as with high quality genomic DNA template from a further 16 unrelated fungi, including Candida albicans, C. dubliniensis, C. parapsilosis, C. glabrata, Scedosporium apiospermum, Penicillium sp., Aspergillus fumigatus, Aspergillus versicolor, Pichia guilliermondii, Rhodotorula sp., Trichosporon sp., Aureobasidium pullulans, Fusarium sp., Mucor hiemalis, Bionectria ochroleuca, Gibberella pulicaris. Results demonstrated that only DNA from E. dermatitidis gave an amplification product of the expected sire, whilst none of the other fungi were amplifiable. Subsequent employment of this primer pair detected this yeast from mycological cultures from 2/50 (4%) adult CF patients. These two patients were the only patients who were previously shown to have a cultural history of E. dermatitidis from their sputum. E. dermatitidis is a slow-growing fungus, which usually takes up to two weeks to culture in the microbiology laboratory and therefore is slow to detect conventionally, with the risk of bacterial overgrowth from common co-habiting pan- and multiresistant bacterial pathogens from sputum. namely Pseudomonas aeruginosa and Burkholderia cepacia complex organisms, hence this species-specific PCR assay may help detect this organism from CF sputum more specifically and rapidly. Overall, employment of this novel assay nay help in the understanding of the occurrence. aetiology and epidemiology of E. dermatitidis, as an emerging fungal agent in patients with CF. (C) 2008 European Cystic Fibrosis Society. Published by Elsevier B.V. All rights reserved.

FT-IR analysis of acid black dye biodegradation using saccharomyces cerevisiae immobilized with treated sugarcane bagasse

Textile industries use large amounts of water in dyeing processes and a wide variety of synthetic dyes. A small concentration of these dyes in the environment can generate highly visible pollution and changes in aquatic ecosystems. Adsorption, biosorption, and biodegradation are the most advantageous dye removal processes. Biodegradation occurs when enzymes produced by certain microorganisms are capable of breaking down the dye molecule. To increase the efficiency of these processes, cell immobilization enables the reuse of the immobilized cells and offers a high degree of mechanical strength, allowing metabolic processes to take place under adverse conditions. The aim of the present study was to investigate the use of Saccharomyces cerevisiae immobilized in activated sugarcane bagasse for the degradation of Acid Black 48 dye in aqueous solutions. For such, sugarcane bagasse was treated with polyethyleneimine (PEI). Concentrations of a 1 % S. cerevisiae suspension were evaluated to determine cell immobilization rates. Once immobilization was established, biodegradation assays for 240 h with free and immobilized yeast in PEI-treated sugarcane bagasse were evaluated by Fourier transform infrared spectrophotometry. The results indicated a probable change in the dye molecule and the possible formation of new metabolites. Thus, S. cerevisiae immobilized in sugarcane bagasse is very attractive for biodegradation processes in the treatment of textile effluents. © 2013 Springer Science+Business Media Dordrecht.

The foraging behaviour of the Black rat (Rattus rattus) within Australian macadamia orchards

The black rat (Rattus rattus) has been shown to be the primary species responsible for causing significant crop losses within the Australian macadamia industry. This species success within macadamia orchards is directly related to the flexibility expressed in its foraging behaviour. In this paper a conceptual foraging model is presented which proposes that the utilisation of resources by rodents within various components of the system is related not only to their relative abundance, but also to predator avoidance behaviour. Nut removal from high predation risk habitats during periods of low resource abundance in low risk compartments of the system is considered an essential behaviour that allows high rodent densities to be maintained throughout the year.