Rapid monitoring of bioaerosols in Urban, Agricultural and Industrial Environments

城市、农业和工业地区排放的废气对当地空气质量的影响日益受到关注。然而，对空气中微生物的特性、分布和丰度的了解仍处于起步阶段。接触传染性细菌和病毒生物体后，会产生有害的健康影响。事实上，生物气溶胶和挥发性有机化合物对人类暴露的风险仍然难以实时量化。人们还日益关注意外或故意向环境释放生物材料及其对人类/动物健康和经济的相关影响。因此，快速检测生物气溶胶是一项迫切的民用和军事需求。2012年，政府发起了一项“布莱克特评估”（blackkett Review），以解决以下问题：“在未来15年内，哪些技术或能力能够实现对各种生物制剂的快速、广域监测？”由于暴露的风险与微生物的浓度和类型直接相关，如果要评估不同环境中感染物种的时空趋势，就需要更清晰的表征、量化和监测方法。目前的监测方法是劳动时间密集型的，昂贵的，在捕获足够数量的生物气溶胶表征的生物材料方面效率低下。此外，目前还没有建立标准化的方案，这往往导致低估微生物负荷的多样性和数量。因此，需要对生物气溶胶材料有更深入的了解。下一代测序（NGS）促进了生物气溶胶材料在精细尺度上的表征。但是，这些方法的使用没有关于使用哪种过滤器的指导方针；采样速率、时间周期或提取方法。此外，简单地描述气溶胶微生物组只是了解不同环境下生物气溶胶中发生的微生物过程的第一步。元转录组学提供了对群落如何响应其环境变化的理解，可能更适合于分析复杂的生物气溶胶群落及其与环境中生物/非生物因素的相互作用。该项目的总体目标是开发一个“生物工具包”，用于对城市、农业和工业环境中的生物气溶胶进行高可信度、广域的生物检测和生物监测。具体而言，该项目旨在开发用于检测生物气溶胶的快速，高通量样品捕获，浓缩和制备的新技术。该项目将结合NGS（变质）和化学标记分析来表征不同环境下的气溶胶微生物群。这将为识别、量化和监测生物气溶胶中的关键病原体提供一种强有力的、具有成本效益的、敏感的方法。我们将使用亚转录组学来鉴定气溶胶微生物组的功能多样性，并为气溶胶样品中支持细菌多样性的过程提供见解。我们将研究功能多样性如何在环境类型和背景下变化，并提供有关总细菌多样性的额外系统发育信息。最终用户受益者的一个主要项目产出将是一个优化的网络系统，用于在城市、农业和工业环境中使用便携式“现场”微型仪器对生物气溶胶（特别是细菌病原体）进行快速和响应性的广域实时监测（即检测、表征和量化）。我们将建立一个微生物挥发性有机标志物数据库，用于快速表征来自不同来源的生物气溶胶。该项目还使人们更好地了解来自不同来源的生物气溶胶对人类接触的影响，这与NERC战略直接相关。

The impact of emissions from urban, agricultural and industrial areas on local air quality is a growing concern. However, understanding the identities, distribution and abundance of airborne microorganisms remains in its infancy. Deleterious health effects can arise following exposure to infective bacterial and viral organisms. Indeed, the risk to human exposure from bioaerosols and volatile organic compounds are still difficult to quantify in real time. There is also growing concern about the accidental or deliberate release of biological materials in the environment and the associated impacts on human/animal health and the economy. Thus, rapid bioaerosol detection is an urgent civilian and military requirement. In 2012, the Government instigated a "Blackett Review" to address the question "Which technologies or capabilities will enable rapid, wide-area surveillance of a broad spectrum of biological agents in the next 15 years?" As the risk of exposure is directly linked to the concentration and type of microorganisms, much clearer characterisation, quantification and monitoring methods are needed if the temporal and spatial trends of infectious species are to be evaluated in different environments. Current monitoring methods are labour-time intensive, expensive and inefficient at capturing sufficient amounts of biomaterial for bioaerosol characterisation. Furthermore, there is currently no standardised protocol established which often leads to an underestimation of the diversity and quantity of microbial load. Thus, a more in-depth understanding of bioaerosol material is required. Next Generation Sequencing (NGS) has facilitated the characterisation of bioaerosol material at the fine-scale. However, such methods are being used with no guidelines as to the type of filter to be used; sampling flow rate, time period, or extraction methods. Furthermore, simply characterising the aerosol microbiome is only the first step to understanding the microbial processes occurring in bioaerosols in different environments. Metatranscriptomics which provides an understanding of how communities respond to changes in their environment may be more appropriate to analysing complex bioaerosol communities and their interactions with biotic/ abiotic factors in the environment.The overall aim of this project is to develop a 'bio-toolkit' for high-confidence, wide-area biodetection and biomonitoring of bioaerosols from urban, agricultural and industrial environments. Specifically, the project aims to develop novel techniques for rapid, high-throughput sample capture, concentration and preparation for detecting bioaerosols. The project will utilise a combination of NGS (metagenetic) and chemical marker analysis to characterise the aerosol microbiome across environments. This will provide a robust, cost-effective, sensitive approach to identify, quantify and monitor key pathogens in bioaerosols. We will use metatranscriptomics to identify the functional diversity of the aerosol microbiome and provide insights into the processes supporting bacterial diversity in aerosol samples. We will examine how functional diversity changes across environment type and context and provide additional phylogenetic information on total bacterial diversity. A major project output for end-user beneficiaries will be an optimised network system for the rapid and responsive wide-area real-time monitoring (i.e. detection, characterisation and quantification) of bioaerosols (specifically bacterial pathogens) across urban, agricultural and industrial environments using portable 'in-field' micro-instrumentation. We will produce a database of microbial volatile organic markers for the rapid characterisation of bioaerosols from different sources. The project also provides a better understanding of the impact of bioaerosols from different sources on human exposure which is of direct relevance to the NERC strategy.