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Aspergillus fumigatus DBM 4057 biofilm formation is inhibited by chitosan, in contrast to baicalein and rhamnolipid

机译:与黄ical素和鼠李糖脂相反,烟曲霉DBM 4057生物膜的形成被壳聚糖抑制

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摘要

The biofilms of filamentous-forming fungi are a novel and still insufficiently understood research topic. We have studied Aspergillus fumigatus, an ubiquitous opportunistic pathogenic fungus, as a representative model for a study of biofilm formation by filamentous fungi and for assessing the potential anti-biofilm activity of natural substances. The activity of antibiotic amphotericin B and selected natural substances: baicalein, chitosan and rhamnolipid was studied. The minimum suspension inhibitory concentrations ( MIC) were determined and the biofilm susceptibility was investigated by determining the metabolic activity of sessile cells ( XTT assay) and total biofilm biomass ( crystal violet staining). Significant time-dependent differences in substances' anti-biofilm activity were observed. Images of A. fumigatus biofilm were obtained by Cellavista automatic light microscope and spinning disc confocal microscopy. Baicalein and rhamnolipid were not found as suitable substances for inhibition of the A. fumigatus biofilm formation, as neither of the substances inhibited the sessile cells metabolic activity or the total biofilm biomass even at tenfold MIC after 48 h. In contrast, chitosan at 10 9 MIC ( 25 mu g mL(-1)), suppressed the biofilm metabolic activity by 90 % and the total biofilm biomass by 80 % even after 72 h of cultivation. Amphotericin B inhibited only 14 % of total biofilm biomass ( crystal violet staining) and 35 % of metabolic activity ( XTT assay) of adherent cells under the same conditions. Our results therefore suggest chitosan as potential alternative for treating A. fumigatus biofilm-associated infections.
机译:丝状真菌的生物膜是一个新颖的,但仍未被充分了解的研究课题。我们已经研究了烟曲霉,一种普遍存在的机会致病真菌,作为研究丝状真菌生物膜形成和评估天然物质潜在抗生物膜活性的代表性模型。研究了抗生素两性霉素B和选定的天然物质:黄ical素,壳聚糖和鼠李糖脂的活性。确定最低悬浮抑制浓度(MIC),并通过确定无柄细胞的代谢活性(XTT分析)和总生物膜生物量(结晶紫染色)研究生物膜敏感性。观察到物质的抗生物膜活性存在明显的时间依赖性差异。通过Cellavista自动光学显微镜和旋转盘共聚焦显微镜获得烟曲霉生物膜的图像。黄ical素和鼠李糖脂没有被发现是抑制烟曲霉生物膜形成的合适物质,因为即使在48小时后,即使在十倍的MIC下,黄ical素和鼠李糖脂都不能抑制无柄细胞的代谢活性或总生物膜生物量。相反,即使在培养72小时后,壳聚糖在10 9 MIC(25μg mL(-1))下的生物膜代谢活性也被抑制90%,总生物膜生物量被抑制80%。在相同条件下,两性霉素B仅抑制总生物膜生物量的14%(结晶紫染色)和35%的粘附细胞代谢活性(XTT分析)。因此,我们的结果表明壳聚糖可以作为治疗烟曲霉生物膜相关感染的潜在替代品。

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