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This study was aimed to evaluate the pattern of cellulase biosynthesis from Aspergillus fumigatus ABK9 under submerged fermentation. Production was increased concomitantly with fungal growth up to 72 h and reached maximum (Xmax –6.72 g/l) with specific growth rate (µmax) of 0.126/h. Highest specific rate of enzyme production (qp) was found at initial medium pH of 5.0 and incubation temperature of 30°C. At the same time, in the presence of 2-deoxy-D-glucose concentration of 0.5 mg/ml, the production of cellulolytic enzymes, viz, carboxymethyl cellulase activity (CMCase), filter paper degrading activity (FPase) and β-glucosidase activity reached maximum of 132.2, 21.3 and 28.9 U/ml, respectively. Cellulase biosynthesis was induced in respect to higher volumetric production rate (Qp), specific rate of enzymes production (qp, U/g biomass/h) and enzyme/biomass yield (YE/X) when grown in carboxymethyl cellulose in comparison to other saccharides as sole carbon source. Induction ratios (IR) of cellulases were between 12.3 and 24.4 in the presence of 1.5% (w/v) CMC in the culture media. The strain was quite resistant to catabolic repression by glucose up to 0.4% (w/v). Cellulases production was greatly influenced in the presence of yeast extract and potassium dihydrogen phosphate (KH₂PO₄) as nitrogen and phosphate sources in the culture media. C/N ratio of 10.0 and C/P ratio of 4.0 proved to be the best for the production of enzyme cocktail. Along with the high production yield, the crude enzymes showed a promising cellulose hydrolyzing efficiency of rice straw, indicating the enzyme could be beneficial for its large scale industrial exploitation.
Many species of microfungi are reported in aquatic ecosystems with different frequency. Their number constantly fluctuates depending on the concentration of environmental and anthropogenic factors. Drinking water, tap and bottled, is essential for the proper functioning of the human body. It is also the main component of food and hence it should be safe for human health and free of contaminants. The mycological purity of tap water in two large cities in the region (Olsztyn and Ostrołęka) and a small village (Gągławki) as well as bottled, medium-mineralized and curative water stored under different conditions were tested. The laboratory investigations followed a pathway applied in diagnostic mycological laboratories. The conducted tests demonstrated that microfungi were found in tap water originating from the cities and in bottled water. The rural water supply system was free from contaminations. Eighteen species of microfungi were identified in tap water from Olsztyn and 9 species in tap water from Ostrołęka. In bottled water, 13 fungal species were detected. Exophiala spinifera and Debaryomyces hansenii were recorded in the water supply systems of both cities, while one common species, i.e. Aspergillus fumigatus, was identified in tap water from Ostrołęka and in bottled water. The conducted studies have significant practical implications, for instance in sanitary and epidemiological water evaluation and in medicine in the context of analysing the quality of drinking water in reference to health resorts and nosocomial infections.
Celem pracy było badanie i określenie przydatności poddanego mutagenizacji szczepu Aspergillus fumigatus do wytwarzania inwertazy i inulinazy w warunkach wgłębnych hodowli wstrząsanych. Wszystkie filtraty pohodowlane pochodzące z hodowli badanych mutantów, w porównaniu z próbą odniesienia, jaką był filtrat pohodowlany pochodzący z hodowli szczepu rodzimego tego grzyba strzępkowego, cechowały się wyższymi aktywnościami enzymatycznymi zarówno zewnątrzkomórkowej inulinazy jak i inwertazy. Aktywność zewnątrzkomórkowej inulinazy, w porównaniu ze szczepem rodzimym, wzrosła o 1,5-4 razy, natomiast aktywność zewnątrzkomórkowej inwertazy wzrosła 1,5-10,5-krotnie. Najaktywniejszym mutantem Aspergillus fumigatus okazał się mutant po szoku oksydacyjnym oraz mutant wyizolowany na α-D-glukozie. Mutant po stresie oksydacyjnym produkował inwertazę o najwyższej aktywności tj. 21,13 μmol/cm3xmin w 8 dobie hodowli, natomiast najwyższa aktywność inulinazy przypadała na 5 dobę hodowli i wynosiła 9,6 μmol/cm3xmin. Najwyższe aktywności inwertazy i inulinazy dla mutanta wyizolowanego na α-D-glukozie wynosiły odpowiednio 11,17 μmol/cm3xmin w 7 dobie hodowli i 5,71 μmol/cm3xmin w 4 dobie hodowli.
The frequency of severe systemic fungal diseases has increased in the last few decades. The clinical use of antibacterial drugs, immunosuppressive agents after organ transplantation, cancer chemotherapy, and advances in surgery are associated with increasing risk of fungal infections. Opportunistic pathogens from the genera Candida and Aspergillus as well as pathogenic fungi from the genus Cryptococcus can invade human organism and may lead to mucosal and skin infections or to deep-seated mycoses of almost all inner organs, especially in immunocompromised patients. Nowadays, there are some effective antifungal agents, but, unfortunately, some of the pathogenic species show increasing resistance. The identification of fungal virulence factors and recognition of mechanisms of pathogenesis may lead to development of new efficient antifungal therapies. This review is focused on major virulence factors of the most common fungal pathogens of humans: Candida albicans, Aspergillus fumigatus and Cryptococcus neoformans. The adherence to host cells and tissues, secretion of hydrolytic enzymes, phenotypic switching and morphological dimorphism contribute to C. albicans virulence. The ability to grow at 37°C, capsule synthesis and melanin formation are important virulence factors of C. neoformans. The putative virulence factors of A. fumigatus include production of pigments, adhesion molecules present on the cell surface and secretion of hydrolytic enzymes and toxins.
Badaniom na aktywność przeciwgrzybiczą wobec trzech patogennych grzybów: Candida albicans, Rhodotorula rubra, Aspergillus fumigatus, poddano wyciągi octanowe z ziela pięciu krajowych gatunków rodziny Dipsacaceae: Dipsacus silvestris L., Knautia arvensis Coult., Scabiosa ochroleuca L., Succisa pratensis Mnch. i Succisella inflexa Beck. Najbardziej aktywne okazały się ekstrakty z Dipsacus silvestris, Succisa pratensis i Succisella inflexa, słabsze działanie wykazała Knautia arvensis, najsłabsze - Scabiosa ochroleuca. Chromatograficznie (HPLC) stwierdzono, że w badanych wyciągach dominują związki poli- fenolowe z przewagą kompleksu flawonoidów, w tym luetoliny i jej 7-glukozydu, apigeniny, kwer- cetyny, kemferolu oraz fenolokwasy: chlorogenowy, protokatechowy, p-kumarowy i p-hydroksy- benzoesowy. Otrzymane wyniki stanowią podstawę do dalszych badań mających na celu izolację i identyfikację związków odpowiedzialnych za powyższe działanie.
W pracy przedstawiono efekt działania różnych stężeń fosforanu mocznika na 6 szczepów grzybów środowiskowych różniących się intensywnością wzrostu, sposobem zarodnikowania i typem tworzonych zarodników (Penicillium italicum, Aspergillus fumigatus, Aspergillus niger, Cladosporium resinae, Mucor hiemalis, Rhizopus nigricans). Fosforan mocznika już w stężeniu 3% w sposób statystycznie istotny ograniczał wzrost masy grzybni wszystkich testowanych grzybów.
The contamination of dried medicinal plants with microscopic fungi has been the subject of many studies. However, no data on extracellular enzyme activities of xerophilic fungi contaminating the plants have been found in the literature. Therefore, the objective of our study was to determine extracellular enzyme profiles of fast-growing xerophilic fungi, i.e. Aspergillus flavus, A. fumigatus, A. melleus, A. nidulans, A. niger, A. parasiticus and Trichothecium roseum isolated from dried medicinal plants from herbal shops in Szczecin, Poland. Solid media and the API ZYM® test were used for measuring enzyme activities. Among the fungi, A. melleus had the highest hydrolytic activity on milk, gelatin, starch, tributyrin, rapeseed oil and biodiesel oil agars, while A. fumigatus showed extremely high stimulation index values on rapeseed oil and biodiesel oil agars. The stimulation index increased during a 5-day incubation period. In the API ZYM® test A. nidulans showed the highest hydrolase activity. Among the hydrolases, ß-glucosidase activity was the highest, followed by acid phosphatase, N-acetyl-ß-glucosaminidase and naphthol-AS-BI-phosphohydrolase activities. The fungi contaminating dried medicinal plants are able to utilize a number of substrates and, therefore, possess high biodeterioration potential. Due to the ability to degrade hydrocarbons, fungal isolates from dried medicinal plants can be used for biotechnological purposes, e.g. in air biofiltration and waste or soil bioremediation.
The majority of mycotoxins produced by Aspergillus fungi are immunosuppressive agents, and their cytotoxicity may impair defense mechanisms in humans. The objective of the study was evaluation of the cytotoxicity of fungi isolated from an environment where inpatients with impaired immunity were present. The materials comprised 57 fungal strains: Aspergillus fumigatus, Aspergillus niger. Aspergillus ochraceus, Aspergillus flavus, Aspergillus versicolor and Aspergillus ustus isolated from hospital rooms in Cracow. The cytotoxicity of all the strains was evaluated using the MTT test (3-(4,5-dimethylthiazol-2-yl) 2,5 diphenyltetrazolium bromide). To emphasize the differences in cytotoxicity among the particular strains, variance analysis (ANOVA) and Tukey's difference test were used. Out of 57 Aspergillus strains tested, 48 (84%) turned out to be cytotoxic. The cytyotoxicity was high (+++) in 21 strains, mainly in A. fumigatus. The least cytotoxic were A. niger fungi, this being statistically significant (p<0,05). To protect a patient from the adverse effects of mycotoxins, not only his or her immunity status should be evaluated but also the presence of fungi in hospital environment and their cytotoxicity should be monitored (possible exposure).
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