Mapping heterotrophic functions to microbial population structures in wastewater resource recovery processes for optimization and prediction of antimicrobial resistance dissemination.

下水道系统和废水资源回收设施 (WRRF) 中的微生物群落通过流入废水驱动的迁移而相互连接。下水道移民如何塑造 WRRF 中的微生物群落仍然存在争议，但它可能受到特定生物过程（例如，需氧与厌氧）和感兴趣群体的代谢功能的调节。这些考虑因素对于开发促进资源回收的新生物工艺非常重要，这些生物工艺越来越多地利用厌氧技术。它们对于预测通过 WRRF 传播抗菌素耐药基因 (ARG) 也具有重要意义。城市废水被认为是抗微生物药物的热点，当前的抗菌素耐药性危机要求修改废水处理设施的运行，这些设施的设计初衷并不是减少抗微生物药物的传播。这些问题是我的研究计划的核心，其长期目标是在了解微生物生态的基础上开发新型废水资源回收生物工艺，并为整个城市废水系统的合理设计和优化提供理论框架。短期内，该提案旨在更详细地研究下水道和 WRRF 生物过程之间的移民界面。将开发和调整多种工具以实现以下短期目标，包括：（1）开发新的合成废水来操纵独立于移民的生物过程微生物群落，（2）使用新的基于流式细胞术的技术来揭示跨移民界面异养生物的代谢功能，（3）使用多重ARG扩增子测序工具来追踪跨界面的特定等位基因序列变体的命运。结果将提供信息来优化随流入废水迁移的微生物群落功能的生物过程，并预测 ARG 等新兴污染物的命运。影响 本工作将培养博士生2名、硕士生2名、本科生5名。他们在废水工程和微生物群落分析/宏基因组学方面的培训将使他们能够很好地进入日益广泛的跨学科劳动力队伍，并在工艺开发和公共卫生领域获得机会。从技术角度来看，这项工作将为评估有关下水道和 WRRF 之间移民界面的许多问题提供平台。 WRRF 行业不仅负责开发更高效的生物工艺，而且还必须满足人们对 WRRF 作为 ARG 传播和重组热点的日益认识。下水道和WRRF 中的微生物群落对于这两个方面都至关重要。在这种情况下，本提案的框架将告知移民过程中发挥作用的机制，并允许更广泛地预测微生物群落对不断变化的 WRRF 程序的反应。

Microbial communities in sewer systems and in the Wastewater Resource Recovery Facilities (WRRFs) are connected through immigration driven by the influent wastewater. How sewer immigration shapes the microbial communities in WRRFs is still debated, but it is likely modulated by the specific bioprocess (e.g., aerobic vs. anaerobic) and the metabolic functions of the populations of interest. These considerations are important for the development of new bioprocesses that promote resource recovery, which increasingly utilize anaerobic technologies. They are also significant for the prediction of the dissemination of antimicrobial resistance genes (ARGs) through WRRFs. Municipal wastewater is recognized as a hotspot of ARGs, and the current antimicrobial resistance crisis demands to revise the operation of wastewater facilities that are not designed to reduce the dissemination of ARGs. These issues are at central to my research program with the long-term objective of developing novel wastewater resource recovery bioprocesses based on an understanding of their microbial ecology and to provide the theoretical framework for the rational design and optimization of the urban wastewater systems as a whole. In the short-term, this proposal aims to look in more details at the immigration interface between sewer and WRRF bioprocesses. Several tools will be developed and adapted to reach the following short-term goals including: (1) the development of new synthetic wastewater to manipulate the bioprocesses microbial community independently of immigration, (2) the use new flow cytometry-based techniques to reveal the metabolic functions heterotrophs across the immigration interface, (3) the use of a multiplex ARG amplicon sequencing tools to trace fate of specific allelic sequence variants across the interface. The results will provide information to optimize bioprocesses in function of the microbial communities immigrating with the influent wastewater, and to predict the fate of emerging pollutants like ARGs. Impact This work will train 2 PhDs, 2 Master's and 5 undergraduate students. Their training in both wastewater engineering and microbial community analyses/metagenomics will equip them well to enter an increasingly broad interdisciplinary workforce with opportunities in process development and public health. From a technical perspective, this work will provide the platform to evaluate a number of questions with respect to the immigration interface between sewers and WRRFs. The WRRF industry is not only charged with the development of more efficient bioprocesses, but also has to content with an increasing awareness of WRRFs as hotspots for the dissemination and recombination of ARGs. Microbial communities in sewers and WRRFs are critical for both these aspects. In this context, the framework of this proposal will inform the mechanism at play during immigration, and allow a wider prediction of microbial community responses to changing WRRF procedures.