Abstract Xylose is the most abundant pentose sugar in lignocellulosic biomass and can be biologically converted into economically important products by microorganisms such as yeasts. This study aimed to characterize the bioconversion of xylose into xylitol and ethanol using four strains of Candida materiae belonging to the Spathaspora clade, which includes yeast species with the potential to convert D-xylose to xylitol and ethanol. Fermentation assays were performed in YPX medium (yeast extract, peptone, xylose 5%) under two oxygen-limiting conditions at inoculum concentrations of 1, 5 and 10 g L-1. The best results were obtained using a 5 g L-1 inoculum under moderate oxygen-limiting conditions. C. materiae UFMG-CM-Y480 produced 32.23 g L-1 of xylitol, yielding 0.743 g g-1 within 36 h. Different combinations of xylose and glucose (25/25 g L-1, 40/10 g L-1, 10/40 g L-1) were also used to verify the influence of sugars on C. materiae metabolism. Xylitol was preferentially produced in medium containing 40 g L-1 xylose. C. materiae UFMG-CM-Y480 produced 26.20 g L-1 of xylitol with a yield of 0.646 g g-1 in 36 h. Our results showed that C. materiae UFMG-CM-Y480 may convert D-xylose to xylitol with a better fermentative performance than other xylitol-producing yeasts.

ABSTRACT Different yeast strains from forests located in southern Brazil, with potential to produce carotenoids, were isolated. Three microorganisms were selected as potential carotenoid producers. Sporiodiobolus pararoseus, Rhodotorula mucilaginosa and Pichia fermentans were grown in Yeast Malt (YM) medium and the carotenoids produced identified as cryptoxanthin and β-carotene. In order to reduce production costs, agroindustrial residues were used in the formulation of medium A (parboiled rice water and crude glycerol) and medium B (parboiled rice water and sugar cane molasses). The highest carotenoid production was obtained with S. pararoseus. It reached 905.30 μ gL-1 (122.82 μg g-1) in YM medium, 820 μg L-1 (68.04 μg g-1) in medium B and 710 μg L-1 (86.46 μg g-1) in medium A. R. mucilaginosa exhibited the best performance in medium B (360 μg L-1 and 30.16 μg g-1) and a new microorganism - P. fermentans - reached 48% (medium A) and 78% (medium B) of the value found in YM medium. Therefore, the agroindustrial residues under evaluation, which replaced the commonly used nitrogen and carbon sources in culture media, enabled the isolated yeasts to yield carotenoids.

BACKGROUND Opportunistic pathogenic yeast species are frequently associated with water habitats that have pollution sources of human or animal origin. Candida albicans has already been suggested as a faecal indicator microorganism for aquatic environments. OBJECTIVES The goal of this study was to investigate the occurrence of C. albicans and other opportunistic yeasts in sand and seawater samples from beaches in Brazil to assess their correlation with Escherichia coli, and to characterise the pathogenic potential of the yeast isolates. METHODS Opportunistic species (yeasts that grow at 37ºC) were isolated from sand and seawater samples from eight beaches in Brazil during the summer and the winter. Opportunistic yeast species were evaluated for their susceptibility to antifungal drugs, virulence factors, and the in vitro and in vivo biofilm formation. Strains were selected to carry out virulence tests using BALB/c mice. FINDINGS Several water samples could be classified as inappropriate for primary contact recreation in relation to E. coli densities. C. albicans was isolated in low densities. Of the 144 opportunistic yeasts evaluated, 61% displayed resistance or dose-dependent sensitivity to at least one tested drug, and 40% produced proteinase. Strains of C. albicans and Kodamaea ohmeri exhibited the highest rates of adhesion to buccal epithelial cells. All the C. albicans strains that were tested were able to undergo morphogenesis and form a biofilm on catheter fragments in both in vitro and in vivo experiments. It was possible to confirm the pathogenic potential of three of these strains during the disseminated infection test. MAIN CONCLUSIONS The identification of opportunistic yeast species in seawater and sand samples from Brazilian beaches suggest a potential risk to the health of people who use these environments for recreational purposes.

ABSTRACT The present study aimed to identify yeasts species isolated from fish feed and the intestinal tracts of tambatinga fish (Colossoma macropomum × Piaractus brachypomum) cultivated in a Brazilian fish farm. Twenty tambatingas and 30 fish feed samples from different brands were acquired from two commercial establishments in the state of Piauí, Brazil. The sampled guts were divided into three equal parts, namely the anterior, medium and posterior portions, totaling 60 samples. Molecular identification was performed by PCR amplification and sequencing of the D1/D2 regions of the large rRNA subunit gene. Sixteen Candida nivariensis strains in the gut were identified. The fish feed samples showed a higher variability of yeast species, with the following species isolated: Hyphopichia burtonii (23.08%), Lodderomyces elongisporus (15.39%) and Trichosporon asahii (15.39%), Candida nivariensis (7.69%), C. orthopsilosis (7.69%), C. parapsilosis (7.69%), Rhodotorula minuta (7.69%), Sterigmatomyces elviae (7.69%), Cryptococcus liquefaciens (7.69%). Yeast found in feed, however, was not always isolated from tambatinga gut. Molecular identification allowed for the isolation of yeast species not previously reported in fish feed and gut samples.

BACKGROUND Endophytic fungi, present mainly in the Ascomycota and Basidiomycota phyla, are associated with different plants and represent important producers of bioactive natural products. Brazil has a rich biodiversity of plant species, including those reported as being endemic. Among the endemic Brazilian plant species, Vellozia gigantea (Velloziaceae) is threatened by extinction and is a promising target to recover endophytic fungi. OBJECTIVE The present study focused on bioprospecting of bioactive compounds of the endophytic fungi associated with V. gigantea, an endemic, ancient, and endangered plant species that occurs only in the rupestrian grasslands of Brazil. METHODS The capability of 285 fungal isolates to produce antimicrobial and antimalarial activities was examined. Fungi were grown at solid-state fermentation to recover their crude extracts in dichloromethane. Bioactive extracts were analysed by chromatographic fractionation and NMR and displayed compounds with antimicrobial, antimycobacterial, and antimalarial activities. FINDINGS Five fungi produced antimicrobial and antimalarial compounds. Extracts of Diaporthe miriciae showed antifungal, antibacterial, and antimalarial activities; Trichoderma effusum displayed selective antibacterial activity against methicillin-resistant Staphylococcus aureus and Mycobacterium intracellulare; and three Penicillium species showed antibacterial activity. D. miriciae extract contained highly functionalised secondary metabolites, yielding the compound epoxycytochalasin H with high antimalarial activity against the chloroquine-resistant strain of Plasmodium falciparum, with an IC50 approximately 3.5-fold lower than that with chloroquine. MAIN CONCLUSION Our results indicate that V. gigantea may represent a microhabitat repository hotspot of potential fungi producers of bioactive compounds and suggest that endophytic fungal communities might be an important biological component contributing to the fitness of the plants living in the rupestrian grassland.

ABSTRACT Fatty acid methyl esters (FAMEs) were obtained from vegetable oils of soybean, corn and sunflower. The current study was focused on evaluating the antifungal activity of FAMEs mainly against Paracoccidioides spp., as well as testing the interaction of these compounds with commercial antifungal drugs and also their antioxidant potential. FAMEs presented small IC50 values (1.86-9.42 μg/mL). All three FAMEs tested showed antifungal activity against isolates of Paracoccidioides spp. with MIC values ranging from 15.6-500 µg/mL. Sunflower FAMEs exhibited antifungal activity that extended also to other genera, with an MIC of 15.6 μg/mL against Candida glabrata and C. krusei and 31.2 μg/mL against C. parapsilosis. FAMEs exhibited a synergetic effect with itraconazole. The antifungal activity of the FAMEs against isolates of Paracoccidioides spp. is likely due to the presence of methyl linoleate, the major compound present in all three FAMEs. The results obtained indicate the potential of FAMEs as sources for antifungal and antioxidant activity.

Fungi of the genus Paracoccidioides are responsible for paracoccidioidomycosis. The occurrence of drug toxicity and relapse in this disease justify the development of new antifungal agents. Compounds extracted from fungal extract have showing antifungal activity. Extracts of 78 fungi isolated from rocks of the Atacama Desert were tested in a microdilution assay against Paracoccidioides brasiliensis Pb18. Approximately 18% (5) of the extracts showed minimum inhibitory concentration (MIC) values≤ 125.0 µg/mL. Among these, extract from the fungus UFMGCB 8030 demonstrated the best results, with an MIC of 15.6 µg/mL. This isolate was identified as Aspergillus felis (by macro and micromorphologies, and internal transcribed spacer, β-tubulin, and ribosomal polymerase II gene analyses) and was grown in five different culture media and extracted with various solvents to optimise its antifungal activity. Potato dextrose agar culture and dichloromethane extraction resulted in an MIC of 1.9 µg/mL against P. brasiliensis and did not show cytotoxicity at the concentrations tested in normal mammalian cell (Vero). This extract was subjected to bioassay-guided fractionation using analytical C18RP-high-performance liquid chromatography (HPLC) and an antifungal assay using P. brasiliensis. Analysis of the active fractions by HPLC-high resolution mass spectrometry allowed us to identify the antifungal agents present in the A. felis extracts cytochalasins. These results reveal the potential of A. felis as a producer of bioactive compounds with antifungal activity.

The aim of the present study was to determine fungal genera, Aspergillus, Pénicillium and Fusarium species and aflatoxin B1 contamination from raw materials and finished feed intended for fish farm localized in Piaui, Brazil. Aspergillusflavus and P. citrinum were isolated with a high relative density from all samples. In general, a high percent of samples exceeded the levels proposed as feed hygienic quality limits (CFU g-1) according to Good Manufacture Practice. Aflatoxin B1 was analyzed by enzyme-linked immunosorbent assay. All raw materials and finished feed showed aflatoxin B1 levels. Although in this study AFB1 levels below recommended limits (20 μg kg-1) were found, it is important to emphasize the feed intake with toxin in low concentrations along time, since it produce chronic deleterious effects in animal production. This fact requires periodic monitoring to prevent the occurrence of chronic aflatoxicosis in aquaculture, to reduce the economic losses and to minimize hazards to animal health.

El objetivo de este estudio fue determinar géneros fúngicos, especies de Aspergillus, Pénicillium y Fusarium y la contaminación de aflatoxina B1 (AFB1) en materias primas y alimento terminado destinado a la cría de peces en el estado de Piauí, Brasil. Aspergillus flavus y P. citrinum fueron aisladas con alta densidad relativa. En general, un alto porcentaje de muestras excedieron los niveles fúngicos propuestos como límite de calidad higiénica (CFU g-1) según las buenas prácticas de manufactura. Aflatoxina B1 fue analizada por ensayos inmuno-enzimáticos. Todas las materias primas y el alimento terminado mostraron niveles de contaminación con AFB1. Aunque en este estudio los niveles de AFB1 estuvieron por debajo del límite recomendado (20 μg de toxina kg-1 de alimento), es importante enfatizar la ingesta de alimento con bajas concentraciones de toxina a lo largo del tiempo ya que produce efectos crónicos en animales de producción. Este factor requiere monitoreo periódico para prevenir la incidencia de aflatoxicosis crónica en acuacultura, reducir las pérdidas económicas y minimizar los riesgos de la salud animal.

Aiming to identify new sources of bioactive secondary metabolites, we isolated 82 endophytic fungi from stems and barks of the native Brazilian tree Caesalpinia echinata Lam. (Fabaceae). We tested their ethyl acetate extracts in several in vitro assays. The organic extracts from three isolates showed antibacterial activity against Staphylococcus aureus and Escherichia coli [minimal inhibitory concentration (MIC) 32-64 μg/mL]. One isolate inhibited the growth of Salmonella typhimurium (MIC 64 μg/mL) and two isolates inhibited the growth of Klebsiella oxytoca (MIC 64 μg/mL), Candida albicans and Candida tropicalis (MIC 64-128 μg/mL). Fourteen extracts at a concentration of 20 μg/mL showed antitumour activities against human breast cancer and human renal cancer cells, while two isolates showed anti-tumour activities against human melanoma cancer cells. Six extracts were able to reduce the proliferation of human peripheral blood mononuclear cells, indicating some degree of selective toxicity. Four isolates were able to inhibit Leishmania (Leishmania) amazonensis and one isolate inhibited Trypanosoma cruzi by at least 40% at 20 μg/mL. The trypanocidal extract obtained from Fusarium sp. [KF611679] culture was subjected to bioguided fractionation, which revealed beauvericin as the compound responsible for the observed toxicity of Fusarium sp. to T. cruzi. This depsipeptide showed a half maximal inhibitory concentration of 1.9 μg/mL (2.43 μM) in a T. cruzi cellular culture assay.

The aims of this work was to characterise indigenous Saccharomyces cerevisiae strains in the naturally fermented juice of grape varieties Cabernet Sauvignon, Grenache, Tempranillo, Sauvignon Blanc and Verdejo used in the São Francisco River Valley, northeastern Brazil. In this study, 155 S. cerevisiae and 60 non-Saccharomyces yeasts were isolated and identified using physiological tests and sequencing of the D1/D2 domains of the large subunit of the rRNA gene. Among the non-Saccharomyces species, Rhodotorula mucilaginosa was the most common species, followed by Pichia kudriavzevii, Candida parapsilosis, Meyerozyma guilliermondii, Wickerhamomyces anomalus, Kloeckera apis, P. manshurica, C. orthopsilosis and C. zemplinina. The population counts of these yeasts ranged among 1.0 to 19 x 10(5) cfu/mL. A total of 155 isolates of S. cerevisiae were compared by mitochondrial DNA restriction analysis, and five molecular mitochondrial DNA restriction profiles were detected. Indigenous strains of S. cerevisiae isolated from grapes of the São Francisco Valley can be further tested as potential starters for wine production.

The ripening process of Serro Minas cheese, one of the most popular cheeses produced with raw milk in Brazil, was studied over the course of 60 days of ripening during dry and rainy seasons. Brazilian legislation prohibits the production of cheese from raw milk unless it was submitted to a maturation period greater than 60 days. However Minas Serro cheese is sold within a few days of ripening. A total of 100 samples of Serro cheese were obtained from five farms; 50 samples were collected during the dry season (winter in Brazil) and 50 samples were collected during the rainy season (summer in Brazil). From each farm, ten cheeses were collected during each season after two days of ripening. Our results showed high levels of total and fecal coliforms at the beginning of the ripening period (approximately 4 Log MPN/g with 3 days of ripening) that decreased with 60 days of ripening reaching almost 1.5 Log MPN/g. Contamination by coagulase-positive staphylococci was reduced by the end of the ripening period. Salmonella spp. was not detected. The staphylococcal enterotoxins B and C were detected in 1% and 4% of the cheeses, respectively, after 30 days of ripening. These results suggest that the ripening process was not effective in eliminating staphylococcal enterotoxins from the cheese. However, none of the investigated strains of Staphylococcus spp. isolated from Serro cheese produced enterotoxins A, B, C or D. The high pathogen and coliform levels at the beginning of the ripening process for the cheese produced during both seasons indicate the need for improvement of the sanitation of the manufacturing conditions.

The aim of this work was to study the yeast populations and the main hygienic-sanitary microbial indicators in water buffalo mozzarella produced and commercialized in Minas Gerais, Brazil. Forty-two water buffalo mozzarella samples were purchased from retail outlets in Belo Horizonte. In addition, five samples of consecutive starter cultures, curd before acidification, acidified curd and mozzarella were collected at an industry in the city of Oliveira. Only three of the five water samples analyzed were suitable for consumption according to Brazilian sanitary standards. Four milk samples were highly contaminated with fecal coliforms, and did not meet the minimal hygienic-sanitary standards according to Brazilian regulations. Only one sample of buffalo muzzarela purchased from retail outlets exceeded the limit for coagulase-positive Staphylococcus. Eleven samples showed counts of thermotolerant coliforms higher than5x 10³ CFU.g-1, but still lower than the maximum permitted by the Brazilian laws. Salmonella spp. and Listeria monocytogenes were not isolated. Debaryomyces hansenii, Candida lusitaniae and C. parapsilosis were the prevalent yeast species isolated from cheese. Among samples from the production stages, the acidified curd presented the highest numbers of yeasts, with C. catenulata being the most frequent species isolated. Some opportunistic yeast species such as C. guilliermondii, C. tropicalis, C. parapsilosis, C. lusitaniae, C. catenulata, C. rugosa and C. krusei occurred in the mozzarella cheese samples analyzed. The mozzarella cheese presented a low microbial load as compared to other cheese already studied, and the yeast biota included species typical of cheese and also opportunistic pathogens.

Yeast communities were assessed in 14 rivers and four lakes from the Doce River basin in Brazil, during the rainy and dry seasons of the years 2000 and 2001. Water samples were collected at the subsurface in all sites. The following physical and chemical parameters were measured: temperature, dissolved oxygen, pH, electrical conductivity, total phosphorus, ortho-phosphate, ammonium, nitrate, nitrite and total nitrogen and the counts of faecal coliforms and heterotrophic bacteria were carried out to characterize the aquatic environmental sampled. The yeast counts were higher in aquatic environments with the highest counts of coliform and heterotrophic bacteria. These environments receive a high influx of domestic and industrial waste. A total of 317 isolates identified in forty eight yeast species were recorded in the sites sampled and the specie Aureobasidium pullulans were found in eleven out of eighteen sites sampled and some opportunistic pathogens such as the yeast species Candida krusei were isolated only in the polluted rivers with a positive correlation with the biotic and abiotic parameters that indicate sewage contamination.

O mecanismo de redução eletroquímica de citrinina em acetonitrila mais (C4H9)4NClO4 0,1 mol L-1 tanto em eletrodo de carbono vítreo (GC) não modificado quanto em eletrodo GC modificado com nanotubos de carbono de paredes múltiplas foi investigado por voltametria cíclica e eletrólise a potencial controlado. Os resultados obtidos permitiram inferir um mecanismo de eletroredução complexo com etapas químicas e eletroquímicas, acopladas à reação inicial de transferência de elétron. Citrinina apresentou pico catódico único que corresponde no mínimo a duas etapas de redução envolvendo um elétron e duas reações químicas homogêneas, segundo mecanismo de autoprotonação do tipo ECEC (transferência de elétron acoplada com aceitação de próton). As reações químicas se originam nas transferências de próton intermolecular do substrato para os seus produtos intermédios básicos de redução, caracterizando um mecanismo de autoprotonação. Análise cinética por procedimentos de simulação de resultados voltamétricos permitiu uma caracterização completa do mecanismo de redução heterogêneo de citrinina.

Electrochemical reduction mechanism of citrinin in acetonitrile plus 0.1 mol L-1 (C4H9)4NClO4 at both bare and modified with multi-walled carbon nanotube glassy carbon (GC) electrodes was investigated by cyclic voltammetry and controlled potential electrolysis. Results allowed to infer a complex electroreduction mechanism with chemical and electrochemical steps coupled to the initial electron transfer reaction. Citrinin shows a single cathodic peak that corresponds at least to two electron reduction steps and two homogeneous chemical reactions, conforming to an ECEC (electron-transfer coupled with proton acceptance) self-protonation mechanism. The chemical reactions are originated in the intermolecular proton transfers from the substrate to its basic reduction intermediates, featuring a self-protonation mechanism. Kinetics analysis by simulation procedures of voltammetric results permitted a fully characterization of the mechanism of citrinin heterogeneous reduction.

The diversity of yeasts collected from different sites in Antarctica (Admiralty Bay, King George Island and Port Foster Bay and Deception Island) and their ability to produce extracellular enzymes and mycosporines were studied. Samples were collected during the austral summer season, between November 2006 and January 2007, from the rhizosphere of Deschampsia antarctica, ornithogenic (penguin guano) soil, soil, marine and lake sediments, marine water and freshwater from lakes. A total of 89 isolates belonging to the following genera were recovered: Bensingtonia, Candida, Cryptococcus, Debaryomyces, Dioszegia, Exophiala, Filobasidium, Issatchenkia (Pichia), Kodamaea, Leucosporidium, Leucosporidiella, Metschnikowia, Nadsonia, Pichia, Rhodotorula, and Sporidiobolus, and the yeast-like fungi Aureobasidium, Leuconeurospora and Microglossum. Cryptococcus victoriae was the most frequently identified species. Several species isolated in our study have been previously reported to be Antarctic psychophilic yeasts, including Cr. antarcticus, Cr. victoriae, Dioszegia hungarica and Leucosporidium scottii. The cosmopolitan yeast species A. pullulans, C. zeylanoides, D. hansenii, I. orientalis, K. ohmeri, P. guilliermondii, Rh. mucilaginosa, and S. salmonicolor were also isolated. Five possible new species were identified. Sixty percent of the yeasts had at least one detectable extracellular enzymatic activity. Cryptococcus antarcticus, D. aurantiaca, D. crocea, D. hungarica, Dioszegia sp., E. xenobiotica, Rh. glaciales, Rh. laryngis, Microglossum sp. 1 and Microglossum sp. 2 produced mycosporines. Of the yeast isolates, 41.7% produced pigments and/or mycosporines and could be considered adapted to survive in Antarctica. Most of the yeasts had extracellular enzymatic activities at 4ºC and 20ºC, indicating that they could be metabolically active in the sampled substrates.