Antifungal Efficacy during Candida krusei Infection in Non-Conventional Models Correlates with the Yeast In Vitro Susceptibility Profile

The incidence of opportunistic fungal infections has increased in recent decades due to the growing proportion of immunocompromised patients in our society. Candida krusei has been described as a causative agent of disseminated fungal infections in susceptible patients. Although its prevalence remains low among yeast infections (2-5%), its intrinsic resistance to fluconazole makes this yeast important from epidemiologic aspects. Non mammalian organisms are feasible models to study fungal virulence and drug efficacy. In this work we have used the lepidopteran Galleria mellonella and the nematode Caenorhabditis elegans as models to assess antifungal efficacy during infection by C. krusei. This yeast killed G. mellonella at 25, 30 and 37°C and reduced haemocytic density. Infected larvae melanized in a dose-dependent manner. Fluconazole did not protect against C. krusei infection, in contrast to amphotericin B, voriconazole or caspofungin. However, the doses of these antifungals required to obtain larvae protection were always higher during C. krusei infection than during C. albicans infection. Similar results were found in the model host C. elegans. Our work demonstrates that non mammalian models are useful tools to investigate in vivo antifungal efficacy and virulence of C. krusei. © 2013 Scorzoni et al.

A comparison between neutron-fluence measurements using metal-activation monitors and standard glasses calibrated via thin uranium-fission monitors and via ηq method

The two fundamental approaches to fission-track dating involve either an explicit determination of the thermal neutron fluence (φ-method) or a calibration against age standards (ζ-method). The neutron fluence measurements are carried out with metal-activation monitors or with uranium-fission monitors, co-irradiated with the samples. Uranium-fission monitors consist of either a thin mono-atomic) film, or a thick fission source (standard uranium glass) irradiated against a muscovite external track detector. In this work, different techniques for performing neutron-fluence measurements were compared: based on thin-film calibration, based on thick-source calibration, and based on gamma spectrometry of co-irradiated metal monitors (Au, Co). The results suggest that more experiments are needed to make all calibrations consistent, including new measurements of the length of etched induced tracks in mica. Also the standard glass calibration carried out with thin films should be confirmed with a greater number of calibrating irradiations. © 2013 Elsevier Ltd. All rights reserved.

Yeast proteins originated from the production of brazilian bioethanol quantification and content

The purpose of this work was to determine the levels of protein and the amino acid distribution in the cell mass of yeast strains (Saccharomyces sensu stricto) originated from Brazilian bioethanol industries. The protein was analyzed with the Kjeldahl method and the amino acids, by using high-performance liquid chromatography (HPLC). The percentages of the protein found ranged from 39 to 49%. The results show that in spite of some variation in numbers between the different yeast strains, all of them presented an amino acid profile similar to the one in the literature for S. cerevisae. The amino acids that have occurred in the largest amounts were: aspartic, glutamic acids and lysine, and those in the lowest amounts were: cysteine and methionine. Although the characteristics of the feedstock used and the process conditions are determinant of the protein values obtained in dry mass, this work elucidates that the intrinsic properties of the yeast strain influence these values.

Evaluation of Rice Bran Extract as a Nitrogen Source for Improved Hemicellulosic Ethanol Production from Sugarcane Bagasse by New Xylose-Fermenting Yeast Strains Isolated from Brazilian Forests

Xylose is the main sugar in hemicellulosic hydrolysates and its fermentation into ethanol by microorganisms is influenced by nutritional factors, such as nitrogen source, vitamins and other elements. Rice bran extract (RBE) is an inexpensive nitrogen source primarily consisting of high amount of protein. This study evaluates the potential of RBE as a nitrogen source for the hemicellulosic ethanol production from sugarcane bagasse dilute acid hydrolysate by novel yeast strains Scheffersomyces shehatae (syn. Candida shehatae) CG8-8BY and Spathaspora arborariae UFMG-HM19.1A, isolated from Brazilian forests. Two different media formulations were used for inoculum preparation and production medium, using yeast extract and RBE as nitrogen sources. S. shehatae CG8-8BY showed ethanol production of 17.0 g/l with the ethanol yield (0.33 g/g) and fermentation efficiency (64 %) from medium supplemented with RBE. On the other hand, S. arborariae presented 5.4 g/l of ethanol production with ethanol yield (0.14 g/g) and fermentation efficiency (21 %) in a fermentation medium supplemented with RBE. Appropriate media formulation is an important parameter to increase the productivity of bioconversion process and RBE proved to be an efficient and inexpensive nitrogen source to supplement sugarcane bagasse hemicellulosic hydrolysate for second generation ethanol production. © 2013 Society for Sugar Research & Promotion.

Effect of plasma and/or yeast extract on performance and intestinal morphology of piglets from 7 to 63 days of age

The objective of this study was to evaluate the effect of diets supplemented with plasma and/or yeast extract on performance (daily weight gain [DWG], daily feed intake [DFI] and feed conversion [FC]) and intestinal morphology of piglets from 7 to 63 days of age. A total of 288 piglets aged 7 days and weighing 2.57±0.05 kg were studied. A randomized block design consisting of four experimental diets, six repetitions and 12 piglets per experimental unit was adopted. The pre-starter I (7 to 21 days), pre-starter II (22 to 35 days) and starter I (36 to 49 days) diets were supplemented as follows: control diet (CD): no plasma or yeast extract; plasma (PL) diet: addition of 6%, 4% and 2% plasma; yeast extract (YE) diet: addition of 6%, 4% and 2% yeast extract; plasma + yeast extract (PL+YE) diet: addition of 3%, 2% and 1% plasma and yeast extract each. From 50 to 63 days of age all piglets received the same diet. No difference in performance was observed from 7 to 21 days and from 7 to 28 days of age, whereas DWG was higher from 7 to 35 days in piglets receiving the PL+YE diet (268, 278, 271 and 288 g/day for CD, PL, YE and PL+YE, respectively). From 7 to 49 days and from 7 to 63 days, DWG (330 and 519 g/day, respectively) and DFI (307 and 647 g/day) were higher in animals receiving the PL-YE diet when compared with those consuming CD (DWG: 295 and 486 g/day; DFI: 266 and 594 g/day). No significant differences in intestinal morphology were observed between piglets receiving the different diets. The combination of plasma and yeast extract elevates DWG, but does not affect the intestinal morphology of piglets from 7 to 63 days of age. © 2013 Sociedade Brasileira de Zootecnia.

Microsatellite marker-based assessment of the biodiversity of native bioethanol yeast strains

Although many Brazilian sugar mills initiate the fermentation process by inoculating selected commercial Saccharomyces cerevisiae strains, the unsterile conditions of the industrial sugar cane ethanol fermentation process permit the constant entry of native yeast strains. Certain of those native strains are better adapted and tend to predominate over the initial strain, which may cause problems during fermentation. In the industrial fermentation process, yeast cells are often exposed to stressful environmental conditions, including prolonged cell recycling, ethanol toxicity and osmotic, oxidative or temperature stress. Little is known about these S. cerevisiae strains, although recent studies have demonstrated that heterogeneous genome architecture is exhibited by some selected well-adapted Brazilian indigenous yeast strains that display high performance in bioethanol fermentation. In this study, 11 microsatellite markers were used to assess the genetic diversity and population structure of the native autochthonous S. cerevisiae strains in various Brazilian sugar mills. The resulting multilocus data were used to build a similarity-based phenetic tree and to perform a Bayesian population structure analysis. The tree revealed the presence of great genetic diversity among the strains, which were arranged according to the place of origin and the collection year. The population structure analysis revealed genotypic differences among populations; in certain populations, these genotypic differences are combined to yield notably genotypically diverse individuals. The high yeast diversity observed among native S. cerevisiae strains provides new insights on the use of autochthonous high-fitness strains with industrial characteristics as starter cultures at bioethanol plants. © 2013 John Wiley & Sons, Ltd.

Further investigation of the initial fission-track length and geometry factor in apatite fission-track thermochronology

The external detector method (EDM) is a widely used technique in fission track thermochronology (FTT) in which two different minerals are concomitantly employed: spontaneous tracks are observed in apatite and induced ones in the muscovite external detector. They show intrinsic differences in detection and etching properties that should be taken into account. In this work, new geometry factor values, g, in apatite, were obtained by directly measuring the ρed/ρis ratios and independently determined [GQR]ed/is values through the measurement of projected lengths. Five mounts, two of which were large area prismatic sections and three samples composed of random-orientation pieces have been used to determine the g-values. A side effect of applying EDM is that the value of the initial confined induced fission track, L0, is not measured in routine analyses. The L 0-value is an important parameter to quantify with good confidence the degree of annealing of the spontaneous fission tracks in unknown-age samples, and is essential for accurate thermal history modeling. The impact of using arbitrary L0-values on the inference of sample thermal history is investigated and discussed. The measurement of the L0-value for each sample to be dated using an extra irradiated apatite mount is proposed. This extra mount can be also used for determining the g value as an extension of the ρed/ρis ratio method. Eight apatite samples from crystalline basement, with grains at random orientation, were used to determine the g-values. The results found are statistically in agreement with the values found for apatite samples (from Durango, Mexico) measured in prismatic section and also measured at random orientation. There was no observable variation in efficiency regarding crystal orientation, showing that it is relatively safe using non-prismatic grains, especially in samples with paucity of grains, as it is the case of most basin samples. Implications for the ζ-calibration and for the calibration of the direct (spectrometer-based) fission-track dating are also discussed.

Yeast diversity isolated from grape musts during spontaneous fermentation from a brazilian winery

Saccharomyces and non-Saccharomyces yeast species from a winery located in Brazil were identified by ribosomal gene-sequencing analysis. A total of 130 yeast strains were isolated from grape surfaces and musts during alcoholic fermentation from Isabel, Bordeaux, and Cabernet Sauvignon varieties. Samples were submitted to PCR-RFLP analysis and genomic sequencing. Thirteen species were identified: Candida quercitrusa, Candida stellata, Cryptococcus flavescens, Cryptococcus laurentii, Hanseniaspora uvarum, Issatchenkia occidentalis, Issatchenkia orientalis, Issatchenkia terricola, Pichia kluyveri, Pichia guilliermondii, Pichia sp., Saccharomyces cerevisiae, and Sporidiobolus pararoseus. A sequential substitution of species during the different stages of fermentation, with a dominance of non-Saccharomyces yeasts at the beginning, and a successive replacement of species by S. cerevisiae strains at the final steps were observed. This is the first report about the yeast distribution present throughout the alcoholic fermentation in a Brazilian winery, providing supportive information for future studies on their contribution to wine quality. © 2013 Springer Science+Business Media New York.

Estrogenic and mutagenic activities of crotalaria pallida measured by recombinant yeast assay and ames test

Background: Crotalaria pallida Ailton is a plant belonging to the Fabaceae family, popularly known as rattle or rattlesnake and used in traditional medicine to treat swelling of the joints and as a vermifuge. Previous pharmacological studies have also reported anti-inflammatory, antimicrobial and antifungal activities. Nevertheless, scientific information regarding this species is scarce, and there are no reports related to its possible estrogenic and mutagenic effects. Thus, the purpose of the present study was to investigate the estrogenic potential of C. pallida leaves by means of the Recombinant Yeast Assay (RYA), seeking an alternative for estrogen replacement therapy during menopause; and to reflect on the safe use of natural products to assess the mutagenic activity of the crude extract from C. pallida leaves, the dichloromethane fraction and stigmasterol by means of the Ames test.Methods: The recombinant yeast assay with the strain BY4741 of Saccharomyces cerevisiae, was performed with the ethanolic extract, dichloromethane fraction and stigmasterol isolated from the leaves of C. pallida. Mutagenic activity was evaluated by the Salmonella/microsome assay (Ames test), using the Salmonella typhimurium tester strains TA100, TA98, TA97 and TA102, with (+S9) and without (-S9) metabolization, by the preincubation method.Results: All samples showed estrogenic activity, mainly stigmasterol. The ethanolic extract from C. pallida leaves showed mutagenic activity in the TA98 strain (-S9), whereas dichloromethane fraction and stigmasterol were found devoid of activity.Conclusion: Considering the excellent estrogenic activity performed by stigmasterol in the RYA associated with the absence of mutagenic activity when evaluated by the Ames test, stigmasterol becomes a strong candidate to be used in hormone replacement therapy during menopause. © 2013 Boldrin et al.; licensee BioMed Central Ltd.

Susceptibility profile of a Brazilian yeast stock collection of Candida species isolated from subjects with Candida-associated denture stomatitis with or without diabetes

Objective: This study investigated the susceptibility of 198 clinical isolates of Candida species against caspofungin, amphotericin B, itraconazole, and fluconazole. Study Design: Suspensions of the microorganisms were spread on Roswell Park Memorial Institute (RPMI) agar plates. Etest strips were placed on the plates, and the minimal inhibitory concentration (MIC) was read after incubation (48 h at 37°C). Data were analyzed by a factorial analysis of variance and a 2 × 2 post hoc test (α = .05). Results: C glabrata showed the highest MIC values (P < .001) against caspofungin, itraconazole, and fluconazole. For amphotericin B, the MIC values of C tropicalis and C glabrata (P = .0521) were higher than those of C albicans (P < .001). Itraconazole was the least effective antifungal; 93.3% of the C glabrata isolates, 3.3% of the C albicans, and 1.3% of the C tropicalis were resistant. All microorganisms were susceptible to caspofungin and amphotericin B. Conclusions: Caspofungin and amphotericin B should be recommended as an effective alternative for the management of oral Candida infections when treatment with topical or other systemic drugs has definitely failed. © 2013 Elsevier Inc. All rights reserved.

Evolution of the South Atlantic passive continental margin in southern Brazil derived from zircon and apatite (U-Th-Sm)/He and fission-track data

The area between São Paulo and Porto Alegre in southeastern Brazil plays a key area to understand and quantify the evolution of the South Atlantic passive continental margin (SAPCM) in Brazil. In this contribution, we present new thermochronological data attained by fission-track and (U-Th-Sm)/He analysis on apatites and zircons from metamorphic, sedimentary and intrusive rocks. The zircon fission-track ages range between 108.4 (15.0) and 539.9 (68.4). Ma, the zircon (U-Th-Sm)/He ages between 72.9 (5.8) and 525.1(2.4). Ma, whereas the apatite fission-track ages range between 40.0 (5.3) and 134.7 (8.0). Ma, and the apatite (U-Th-Sm)/He ages between 32.1 (1.5) and 93.0 (2.5). Ma. The spatial distribution of these ages shows three distinct blocks with a different evolution cut by old fracture zones. While the central block exhibits an old stable block, the Northern and especially the Southern block underwent complex post-rift exhumation. The sample of the Northern block shows two distinct cooling phases in the Upper Cretaceous and the Paleogene to Neogene. After sedimentation of the Permian sandstones the samples of the Central block were never heated up over 100. °C with a following moderate to fast cooling phase in Cretaceous to Eocene time and a fast cooling between Oligocene to Miocene. The five thermal models obtained in the Southern block indicate a complex evolution with three cooling phases. The exhumation events of the three blocks correspond with the Paraná-Etendekka event, the alkaline intrusions due to the Trinidad hotspot, and the evolution of the continental rift basins in SE Brazil and are, therefore, most likely to be the major force for the post-rift evolution of the passive continental margin in SE Brazil, which therefore corresponds to the three main phases of the Andean orogeny. © 2013 Elsevier B.V.

Evaluation of Estrogenic Potential of Flavonoids Using a Recombinant Yeast Strain and MCF7/BUS Cell Proliferation Assay

Phytoestrogens are of interest because of their reported beneficial effects on many human maladies including cancer, neurodegeneration, cardiovascular disease and diabetes. Furthermore, there is a search for compounds with estrogenic activity that can replace estrogen in hormone replacement therapy during menopause, without the undesirable effects of estrogen, such as the elevation of breast cancer occurrence. Thus, the principal objective of this study was to assess the estrogenic activity of flavonoids with different hydroxylation patterns: quercetin, kaempferol, luteolin, fisetin, chrysin, galangin, flavone, 3-hydroxyflavone, 5-hydroxyflavone and 7-hydroxyflavone via two different in vitro assays, the recombinant yeast assay (RYA) and the MCF-7 proliferation assay (E-screen), since the most potent phytoestrogens are members of the flavonoid family. In these assays, kaempferol was the only compound that showed ERα-dependent transcriptional activation activity by RYA, showing 6.74±1.7 nM EEQ, besides acting as a full agonist for the stimulation of proliferation of MCF-7/BUS cells. The other compounds did not show detectable levels of interaction with ER under the conditions used in the RYA. However, in the E-screen assay, compounds such as galangin, luteolin and fisetin also stimulated the proliferation of MCF-7/BUS cells, acting as partial agonists. In the evaluation of antiestrogenicity, the compounds quercetin, chrysin and 3-hydroxyflavone significantly inhibited the cell proliferation induced by 17-β-estradiol in the E-screen assay, indicating that these compounds may act as estrogen receptor antagonists. Overall, it became clear in the assay results that the estrogenic activity of flavonoids was affected by small structural differences such as the number of hydroxyl groups, especially those on the B ring of the flavonoid. © 2013 Resende et al.

Starmerella aceti f.a., sp nova, an ascomycetous yeast species isolated from fungus garden of the leafcutter ant Acromyrmex balzani

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

An automatic method for segmentation of fission tracks in epidote crystal photomicrographs

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