Methods for visualising active microbial benzene degraders in in situ microcosms
Methods for visualising active microbial benzene degraders in in situ microcosms
复制标题
DOI：
10.1007/s00253-014-6037-4
复制
发表时间：
2014
影响因子：
5
通讯作者：
C. Schurig;C. Mueller;C. Höschen;A. Prager;E. Kothe;Henrike Beck;A. Miltner;M. Kästner
中科院分区：
文献类型：
--
作者：
C. Schurig;C. Mueller;C. Höschen;A. Prager;E. Kothe;Henrike Beck;A. Miltner;M. Kästner
自然衰减可能是污染场地生物修复的一个具有成本效益的选择，但需要了解现场条件下降解微生物的活动。为了将微生物活性与污染物降解菌的空间分布联系起来，我们结合了最近改进的原位微宇宙方法，即所谓的“直接推动细菌陷阱”（DP-BACTRAP），并对污染人工湿地样品进行了纳米级二次离子质谱（NanoSIMS）分析。这种方法是基于最初无菌的微观世界，用13 C标记的苯作为微生物的碳源和能源。微宇宙被直接引入人工湿地，在那里它们被沉积物中的土著微生物定殖。在田间培养后，通过NanoSIMS、扫描电子显微镜（SEM）和荧光显微镜对样品进行分析，以便在来自微观世界的未受干扰的样品上观察13 C标记的微生物生物量。随着开发的方法，我们成功地可视化苯降解微生物的固体材料具有高表面积通过NanoSIMS。此外，我们可以证明NanoSIMS分析具有高度复杂地形的未嵌入多孔介质的可行性，这通常被认为不会导致足够的结果。
Natural attenuation maybe a cost-efficient option for bioremediation of contaminated sites but requires knowledge about the activity of degrading microbes under in situ conditions. In order to link microbial activity to the spatial distribution of contaminant degraders, we combined the recently improved in situ microcosm approach, so-called ‘direct-push bacterial trap’ (DP-BACTRAP), with nano-scale secondary ion mass spectrometry (NanoSIMS) analysis on samples from contaminated constructed wetlands. This approach is based on initially sterile microcosms amended with13C-labelled benzene as a source of carbon and energy for microorganisms. The microcosms were introduced directly in the constructed wetland, where they were colonised by indigenous microorganisms from the sediment. After incubation in the field, the samples were analysed by NanoSIMS, scanning electron microscopy (SEM) and fluorescence microscopy in order to visualise13C-labelled microbial biomass on undisturbed samples from the microcosms. With the approach developed, we successfully visualised benzene-degrading microbes on solid materials with high surface area by means of NanoSIMS. Moreover, we could demonstrate the feasibility of NanoSIMS analysis of unembedded porous media with a highly complex topography, which was frequently reasoned to not lead to sufficient results.