Sporotrichosis is a subacute or chronic mycosis caused worldwide by the dimorphic species complex, Sporothrix schenckii. We studied 85 isolates recovered in Brazil to verify their identification and evaluate their in vitro antifungal susceptibility patterns. Based on phenotypic tests (microscopic features, ability to grow at 30°C and 37°C, colony diameters, as well as assimilation of sucrose and raffinose) and molecular assays (amplification of a fragment of the calmodulin gene), the strains were identified as S. schenckii, S. brasiliensis and S. globosa, with a predominance of S. schenckii isolates. There was 37.7% disagreement between the phenotypic and genotypic identification methodologies. In general, terbinafine was the most active drug, followed by ketoconazole and itraconazole, and the less active fluconazole and voriconazole. Five isolates (one S. globosa and four S. schenckii) were found to be itraconazole-resistant strains but, in general, there were no differences in the in vitro antifungal susceptibility profiles among the Sporothrix species.
Zygosaccharomyces bailii BCV 08, a yeast isolated from red wine barrels in Brazil, was evaluated as co-starter in fermentations with Saccharomyces cerevisiae. Z. bailii BCV 08 was preliminarily shown to produce high levels of esters, and the production was optimized in bench and bioreactor scales using grape must. White wine vinifications were conducted with mixed cultures containing different proportions of Z. bailii BCV 08 and an enological strain of S. cerevisiae. In all trials that contained Z. bailii BCV 08, the production of ethyl esters was enhanced in comparison to the vinification control. Our results clearly show the potential of Z. bailii BCV 08 as a mixed starter with S. cerevisiae in order to increase the aromatic complexity of wine.
We report a case of an 80-year-old Brazilian man, farmer, with lesions on the dorsum of the hand. A direct mycological examination, cultivation and microculture slide observation was performed. The sequencing of ITS1-5.8S rDNA-ITS2 region was carried out and the etiological agent confirmed as Exophiala spinifera. The in vitro susceptibility of this isolate to antifungal agents alone and in combination was evaluated. This is the third case of phaeohyphomycosis caused by Exophiala spinifera in Brazil.
Propriedades funcionais-digestivas e nutricionais da polpa-refinada-desidratada de maçã (PRM) foram avaliadas em ratos Wistar, tendo como padrão de fibra o farelo de trigo (FT). A PRM apresentou 91,91% de fibra alimentar/dietária, mais que o dobro do FT (43,69%), sendo 82,27% de fração insolúvel e 9,64% da fração solúvel; 8,20% de proteínas; 0,57% de lipídeos; 2,04% de cinzas (minerais); e ausência de carboidratos digestíveis. A incorporação na dieta dos ratos de PRM ou do padrão FT em proporções de 5%, 15% ou 25% produziu, nos animais, efeitos funcionais-digestivos e nutricionais próprios de fibra alimentar insolúvel. Na concentração de 5% (PRM e FT forneceram 4,6% e 2,2% de fibra alimentar, respectivamente) as duas fontes de fibra produziram, com exceção da densidade de fezes seca, efeitos semelhantes. No entanto, em concentrações de 15% ou 25%, a PRM resultou em mais defecações, maior peso de fezes seca e densidade, porém, produziu fezes seca de menor volume. Com base no resultado da pesquisa, bem como na quantidade industrial disponível, conclui-se que a "polpa-refinada" de maçã poderá constituir-se em fonte alternativa potencial de fibra alimentar para a formulação de alimentos para consumo em dieta normal, mas, principalmente para alimentos especiais que devem apresentar propriedades funcionais-digestivas relacionadas à fibra alimentar.
In microbiology, identification of all isolates by sequencing is still unfeasible in small research laboratories. Therefore, many yeast diversity studies follow a screening procedure consisting of clustering the yeast isolates using MSP-PCR fingerprinting, followed by identification of one or a few selected representatives of each cluster by sequencing. Although this procedure has been widely applied in the literature, it has not been properly validated. We evaluated a standardized protocol using MSP-PCR fingerprinting with the primers (GTG)5 and M13 for the discrimination of wine associated yeasts in South Brazil. Two datasets were used: yeasts isolated from bottled wines and vineyard environments. We compared the discriminatory power of both primers in a subset of 16 strains, choosing the primer (GTG)5 for further evaluation. Afterwards, we applied this technique to 245 strains, and compared the results with the identification obtained by partial sequencing of the LSU rRNA gene, considered as the gold standard. An array matrix was constructed for each dataset and used as input for clustering with two methods (hierarchical dendrograms and QAPGrid layout). For both yeast datasets, unrelated species were clustered in the same group. The sensitivity score of (GTG)5 MSP-PCR fingerprinting was high, but specificity was low. As a conclusion, the yeast diversity inferred in several previous studies may have been underestimated and some isolates were probably misidentified due to the compliance to this screening procedure.
Chromoblastomycosis is a chronic cutaneous and subcutaneous mycosis. The management of this infection continues to be challenging because there is no consensus on the therapeutic regimen. We report here a case of a 69-year-old male patient with cauliflower-like lesions on his left leg and foot. He had already been treated with itraconazole at a dose of 200 mg/day for 5 months, with mycological cure for all the affected areas. However, the lesions relapsed at both sites, and treatment with itraconazole was resumed at the dose previously used. Initially, direct mycological examination, cultural, and microculture slide observation were performed. Afterward, sequencing of the ITS1-5.8S rDNA-ITS2 region of the fungal DNA and evaluation of its susceptibility to antifungal agents alone and in combination were performed. In direct mycological examination, the presence of sclerotic cells was verified, and the fungus was identified as Fonsecaea based on cultural and microscopic examinations. Identification as Fonsecaea monophora was confirmed after sequencing of the ITS region and phylogenetic analysis. The isolate was susceptible to itraconazole and terbinafine. The combinations of amphotericin B and terbinafine and terbinafine and voriconazole were synergistic. The use of drugs for which the causative agent is susceptible to singly or in combination may be an alternative for the treatment of mycosis. Furthermore, the identification of the agent by molecular techniques is important for epidemiological purposes. To the best of our knowledge, this is the first case of relapsed chromoblastomycosis caused by F. monophora in Brazil.
The occurence of Listeria species in refrigerated chicken carcasses was evaluated, comparing the conventional culture methodology of FDA, modified by the introduction of a secondary enrichment step prior plating, and the Clearview TM rapid method (Oxoid, UK Ltd). Forty-eight refrigerated whole chicken carcasses purchased from supermarkets in Florianópolis, SC, Brazil, were analysed. Listeria species occured in 21 (43.7%) samples. Using the Clearview method, 17 (35.4%) samples were positive for Listeria species. Of these isolates, 11 (23%) were L. monocytogenes, 4 (8.3%) L. innocua, 1 (2.1%) L. welshimeri and 1 (2.1%) L. seeligeri. Using the conventional culture methodology of FDA (with modifications), 14 (29.2%) samples were positive for Listeria species. Among these 7 (14.6%) were L. monocytogenes, 6 (12.5%) L. innocua and 1 (2.1%) L. seeligeri. With the Clearview rapid method plus API Listeria for identification, results were confirmed to Listeria species level within 115-139 h. Using the conventional culture method of FDA (with modifications) plus API Listeria, results were confirmed within 120-160 h. However, the Clearview method could indicate the presence of Listeria organisms in only 43 h. Results given by the methods were in moderate concordance and the differences between them were not significant (C.I. = 95%).
Chromoblastomycosis (CBM) is a chronic cutaneous and subcutaneous infection caused by melanized fungal species. We quantified the extractable melanin of 77 strains of CBM agents distributed within five genera. Moreover, resistance to oxidative stress was evaluated in strains exposed or not to the melanin inhibitor tricyclazole. The median percentage of melanin mass extracted from dry fungal mass varied from 0.69 (Rhinocladiella similis) to 3.81 (Phialophora americana). Inhibition of melanin synthesis decreased survival rates to hydrogen peroxide. Together, these data highlight the importance of melanin in CBM agents.
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