Federal Institute for Occupational Safety and Health, Nöldnerstrasse 40-42, 10317 Berlin, Germany.
Division of Microbiology and Animal Hygiene, Department of Animal Sciences, University of Goettingen, Burckhardtweg 2, 37077 Göttingen, Germany.
Ann Work Expo Health. 2017 Jul 1;61(6):633-642. doi: 10.1093/annweh/wxx038.
It is already known that occupational exposure to bioaerosols or organic dust could be harmful for occupants, but mostly the correlation to occurring bacteria is missing. Especially, cultivation of bacteria from bioaerosols is important to get an insight on occurring and possibly infectious bacteria. These measures are highly time consuming and cost intensive. Therefore, to monitor bacterial diversity in bioaerosol samples and to avoid isolation procedures, an approach was applied using a combination of cultivation and cloning-based approach. Preliminary validation of the method was determined using 11 different bacterial strains. After DNA extraction and 16S rRNA gene amplification of grown colonies, subsequent cloning and sequencing was conducted. Initially, to figure out a suitable DNA extraction method, applicable for different airborne bacteria, four DNA extraction protocols were compared. Significantly, best results were determined using the FAST DNA®Spin Kit for Soil with respect to DNA quantity and quality of bacterial cultures. Cloning approach from a mixture of amplified 16S rRNA genes of 11 isolates and following sequence data analysis shows a recovery of all strains when five clones per bacterial strain were analysed. The results clearly demonstrate that a combination of cultivation-based approaches and cloning processes can simplify bioaerosol monitoring of viable and probably infectious bacteria. The implementation of the present method into practice allows a simple and preventive investigation of bioaerosols at work places.
已知职业性接触生物气溶胶或有机粉尘可能对从业人员有害，但发生的细菌与这一情况的相关性大多尚未可知。特别是，从生物气溶胶中培养细菌对于了解存在的和可能具有传染性的细菌非常重要。这些措施非常耗时且耗费成本。因此，为了监测生物气溶胶样本中的细菌多样性并避免分离程序，采用了一种结合培养和基于克隆的方法。使用 11 种不同的细菌菌株对该方法进行了初步验证。在对生长菌落进行 DNA 提取和 16S rRNA 基因扩增后，进行了随后的克隆和测序。最初，为了确定适用于不同空气传播细菌的合适 DNA 提取方法，比较了四种 DNA 提取方案。使用 FAST DNA®Spin Kit for Soil 提取 DNA 时，细菌培养物的 DNA 数量和质量明显最佳。对 11 个分离株的扩增 16S rRNA 基因混合物进行克隆方法处理，并对随后的序列数据分析显示，当每个细菌株系分析 5 个克隆时，可回收所有菌株。结果清楚地表明，培养为基础的方法和克隆过程的结合可以简化对生物气溶胶中存活的和可能具有传染性的细菌的监测。本方法在实践中的实施允许在工作场所对生物气溶胶进行简单的预防性调查。
