| A Reliable Assay to Evaluate the Virulence of Aspergillus nidulans Using the Alternative Animal Model Galleria mellonella (Lepidoptera)
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Author:
Date:
2017-06-05
[Abstract] The greater wax moth Galleria mellonella has emerged as an effective heterologous host to study fungal pathogenesis and the efficacy of promising antifungal drugs (Mylonakis et al., 2005; Li et al., 2013). Here, a methodology describing the Aspergillus nidulans infection in G. mellonella larvae, along with insect survival analysis, is reported. This protocol allowed the distinction between virulent A. nidulans strains (such as TNO2A3), which induced high larval mortality rates, to those in which gene deletion was accompanied by reduced pathogenicity such as ∆gcsA and ∆sdeA (Fernandes et al., 2016).
[摘要] 作为一种有效的异源宿主，越来越多的蜡蛾已经出现了一种有效的异源宿主，用于研究真菌发病机制和有希望的抗真菌药物的功效（Mylonakis等人，2005; Li＆ et al。2013）。这里，描述了一种描述构巢曲霉感染的方法。 mellonella 幼虫与昆虫存活分析一起报道。该协议允许区分有毒的 A。造成高幼虫死亡率的构巢组织菌株（如TNO2A3）与基因缺失伴随着致病性降低的病例如ΔGCA和ΔSAA（Fernandes 等人，2016）。
背景 -G。 mellonella 是一种廉价的模型，易于处理，其先天免疫反应与哺乳动物免疫系统分享功能相似性。此外，感染真菌突变菌株的幼虫和小鼠表现出相似的存活率（Brennan等人，2002）。因此，幼虫构成了一种方便的动物宿主，用于在真菌发病机理分析中代替脊椎动物的使用。尽管昆虫模型具有所有优点，但只有少数报告显示了G中曲霉菌感染的作用。蜡螟。该协议描述了一种有效的方法，用于分析G中的构巢曲霉发病机制。 mellonella 幼虫。
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| Preparation of Candida albicans Biofilms for Transmission Electron Microscopy
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Author:
Date:
2013-07-20
[Abstract] Transmission Electron Microscopy is a form of microscopy that allows for imaging of distinct portions of an individual cell. For Candida albicans biofilms, it is often used to visualize the cell walls of fixed samples of yeast and hyphae. This protocol describes how to grow, harvest, and fix Candida albicans biofilms in preparation for Transmission Electron Microscopy.
[摘要] 透射电子显微术是允许对单个细胞的不同部分成像的显微镜检查的形式。 对于白色念珠菌生物膜，它通常用于可视化酵母和菌丝的固定样品的细胞壁。 此协议描述如何生长，收获和修复白色念珠菌生物膜准备透射电子显微镜。
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| Preparation of Candida albicans Biofilms Using an in vivo Rat Central Venous Catheter Model
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Author:
Date:
2013-07-20
[Abstract] In vivo biofilms grown on medical devices are necessary to understand the interactions of the fungal biofilm and the host environment in which it is most commonly found. This protocol describes a way to grow Candida albicans biofilms on the interior lumen of central venous catheters surgically implanted into rats, which mimics quite well the clinical cases of biofilms found on human central venous catheters. These infected catheters can then be studied via a multitude of different experiments, including cell counting by plating, imaging the catheters under light or electron microscopy, or comparing the relative content of in vivo biofilms to in vitro biofilms and planktonic cultures. These biofilms also provide enough high quality RNA for ...
[摘要] 在医疗装置上生长的体内生物膜是必要的，以了解真菌生物膜和其中最常见的宿主环境的相互作用。 该协议描述了在手术植入大鼠中的中心静脉导管的内腔上生长白假丝酵母生物膜的方法，其相当好地模拟在人中心静脉导管上发现的生物膜的临床情况。 然后可以通过多种不同的实验研究这些感染的导管，包括通过电镀的细胞计数，在光学或电子显微镜下对导管成像，或者将体内生物膜的相对含量与 体外生物膜和浮游培养物。 这些生物膜还提供足够高质量的RNA进行转录分析。
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