Núñez Andrés, Amo de Paz Guillermo, Ferencova Zuzana, Rastrojo Alberto, Guantes Raúl, García Ana M, Alcamí Antonio, Gutiérrez-Bustillo A Montserrat, Moreno Diego A
Escuela Técnica Superior de Ingenieros Industriales, Universidad Politécnica de Madrid (ETSII-UPM), Madrid, Spain.
Departamento de Biología Vegetal II, Universidad Complutense de Madrid (UCM), Madrid, Spain.
Appl Environ Microbiol. 2017 Jun 16;83(13). doi: 10.1128/AEM.00472-17. Print 2017 Jul 1.
Pollen, fungi, and bacteria are the main microscopic biological entities present in outdoor air, causing allergy symptoms and disease transmission and having a significant role in atmosphere dynamics. Despite their relevance, a method for monitoring simultaneously these biological particles in metropolitan environments has not yet been developed. Here, we assessed the use of the Hirst-type spore trap to characterize the global airborne biota by high-throughput DNA sequencing, selecting regions of the 16S rRNA gene and internal transcribed spacer for the taxonomic assignment. We showed that aerobiological communities are well represented by this approach. The operational taxonomic units (OTUs) of two traps working synchronically compiled >87% of the total relative abundance for bacterial diversity collected in each sampler, >89% for fungi, and >97% for pollen. We found a good correspondence between traditional characterization by microscopy and genetic identification, obtaining more-accurate taxonomic assignments and detecting a greater diversity using the latter. We also demonstrated that DNA sequencing accurately detects differences in biodiversity between samples. We concluded that high-throughput DNA sequencing applied to aerobiological samples obtained with Hirst spore traps provides reliable results and can be easily implemented for monitoring prokaryotic and eukaryotic entities present in the air of urban areas. Detection, monitoring, and characterization of the wide diversity of biological entities present in the air are difficult tasks that require time and expertise in different disciplines. We have evaluated the use of the Hirst spore trap (an instrument broadly employed in aerobiological studies) to detect and identify these organisms by DNA-based analyses. Our results showed a consistent collection of DNA and a good concordance with traditional methods for identification, suggesting that these devices can be used as a tool for continuous monitoring of the airborne biodiversity, improving taxonomic resolution and characterization together. They are also suitable for acquiring novel DNA amplicon-based information in order to gain a better understanding of the biological particles present in a scarcely known environment such as the air.
花粉、真菌和细菌是室外空气中存在的主要微观生物实体，会引发过敏症状并传播疾病，在大气动态变化中发挥着重要作用。尽管它们很重要，但尚未开发出一种在大都市环境中同时监测这些生物颗粒的方法。在此，我们评估了使用赫斯特型孢子捕捉器，通过高通量DNA测序来表征全球空气传播生物群，选择16S rRNA基因区域和内部转录间隔区进行分类鉴定。我们表明，这种方法能很好地代表空气生物学群落。两个同步工作的捕捉器的操作分类单元（OTU），在每个采样器收集的细菌多样性中占总相对丰度的>87%，真菌为>89%，花粉为>97%。我们发现显微镜传统表征与基因鉴定之间有良好的对应关系，通过后者获得了更准确的分类鉴定，并检测到了更丰富的多样性。我们还证明，DNA测序能准确检测样本间生物多样性的差异。我们得出结论，应用于用赫斯特孢子捕捉器获得的空气生物学样本的高通量DNA测序能提供可靠结果，并且可以很容易地用于监测城市地区空气中存在的原核生物和真核生物实体。检测、监测和表征空气中存在的广泛多样的生物实体是困难的任务，需要不同学科的时间和专业知识。我们评估了使用赫斯特孢子捕捉器（一种在空气生物学研究中广泛使用的仪器），通过基于DNA的分析来检测和识别这些生物。我们的结果显示DNA收集一致，与传统鉴定方法有良好的一致性，这表明这些设备可作为持续监测空气传播生物多样性的工具，同时提高分类分辨率和表征能力。它们也适用于获取基于新型DNA扩增子的信息，以便更好地了解像空气这样鲜为人知的环境中存在的生物颗粒。
