Gao Xin-Lei, Shao Ming-Fei, Xu Yi-Sheng, Luo Yi, Zhang Kai, Ouyang Feng, Li Ji
Shenzhen Graduate School, State Key Laboratory of Urban Water Resource and Environment, Harbin Institute of TechnologyShenzhen, China; Shenzhen Key Laboratory of Water Resource Utilization and Environmental Pollution ControlShenzhen, China.
State-Key Laboratory of Chemical Engineering, East China University of Science and Technology Shanghai, China.
Front Microbiol. 2016 Dec 1;7:1891. doi: 10.3389/fmicb.2016.01891. eCollection 2016.
Recovering microorganisms from environmental samples is a crucial primary step for understanding microbial communities using molecular ecological approaches. It is often challenging to harvest microorganisms both efficiently and unselectively, guaranteeing a similar microbial composition between original and separated biomasses. A magnetic nanoparticles (MNPs) based method was developed to effectively separate microbial biomass from glass fiber pulp entrapped bacteria. Buffering pH and nanoparticle silica encapsulation significantly affected both biomass recovery and microbial selectivity. Under optimized conditions (using citric acid coated FeO, buffering pH = 2.2), the method was applied in the pretreatment of total suspended particle sampler collected bioaerosols, the effective volume for DNA extraction was increased 10-folds, and the overall method detection limit of microbial contaminants in bioaerosols significantly decreased. A consistent recovery of the majority of airborne bacterial populations was demonstrated by in-depth comparison of microbial composition using 16S rRNA gene high-throughput sequencing. Surface charge was shown as the deciding factor for the interaction between MNPs and microorganisms, which helps developing materials with high microbial selectivity. To our knowledge, this study is the first report using MNPs to separate diverse microbial community unselectively from a complex environmental matrix. The technique is convenient and sensitive, as well as feasible to apply in monitoring of microbial transport and other related fields.
从环境样本中回收微生物是使用分子生态学方法理解微生物群落的关键第一步。高效且无选择性地收获微生物通常具有挑战性,要确保原始生物量和分离后的生物量之间具有相似的微生物组成。开发了一种基于磁性纳米颗粒(MNPs)的方法,以有效地从玻璃纤维纸浆包裹的细菌中分离微生物生物量。缓冲pH值和纳米颗粒二氧化硅包封对生物量回收和微生物选择性均有显著影响。在优化条件下(使用柠檬酸包覆的FeO,缓冲pH = 2.2),该方法应用于总悬浮颗粒采样器收集的生物气溶胶的预处理,DNA提取的有效体积增加了10倍,生物气溶胶中微生物污染物的整体方法检测限显著降低。通过使用16S rRNA基因高通量测序对微生物组成进行深入比较,证明了大多数空气传播细菌种群的一致回收率。表面电荷被证明是MNPs与微生物之间相互作用的决定性因素,这有助于开发具有高微生物选择性的材料。据我们所知,本研究是首次使用MNPs从复杂环境基质中无选择性地分离不同微生物群落的报告。该技术方便、灵敏,并且在微生物运输监测和其他相关领域的应用中也是可行的。