Antibiotic resistance gene propagation: in situ rates and networks of horizontal gene transfer in wastewater

抗生素抗性基因传播：废水中水平基因转移的原位速率和网络

• 批准号: 1805901
• 负责人: Lauren Stadler
• 金额: $ 33万
• 依托单位: William Marsh Rice University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-15 至 2022-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1805901&HistoricalAwards=false
• 关键词: Antibiotic; resistance; gene; propagation; situ

A major challenge to combating the global health crisis of antibacterial resistance is that antibiotic resistance genes (ARGs) can be shared among bacteria through a process called horizontal gene transfer (HGT). ARGs are present in wastewater, and there is little understanding of ARG health impacts potentially conveyed by recycling wastewater. Little is known regarding the conditions that result in HGT of ARGs from environmental bacteria such as those used to treat wastewater and pathogenic bacteria found in clinical infections. The goals of this project are to decipher how pathogens acquire ARGs from environmental bacteria present in water and wastewater systems and to understand how ARGs are propagated in water and wastewater microbial communities. This research will develop biosensors to monitor HGT of ARGs to understand how simple operational parameters in a wastewater treatment plant impact the reduction or proliferation of ARGs. If successful, the results of this project will identify methods of antibiotic resistance transfer in the environment and identify ways to halt this transfer during water treatment processes, protecting public health and the Nation's water supply.Understanding the controls over horizontal gene transfer (HGT) in microbial communities found in the environment would impart an unprecedented ability to manage the growing threat of antibiotic resistance. While a variety of technologies are available for obtaining static snapshots of bacteria that have acquired antibiotic resistance genes (ARGs) through HGT, these existing approaches do not provide dynamic information on the pathways and rates of gene flow within complex communities that experience a changing environment. These approaches also cannot easily differentiate between living and dead bacteria. Two emerging tools will be leveraged to obtain this information: (1) gas-reporting biosensors that report on in situ conjugation events; and (2) a high-throughput, culture-independent method for determining the host-range of ARGs in a mixed community. These tools will be applied in bioreactors treating domestic wastewater to better understand how operational controls impact ARG propagation rates and host range. The objectives of this research are to (1) develop biosensors that report on HGT in situ by coupling the synthesis of an enzyme that produces a rare volatile gas to broad-range plasmid transfer; (2) use these tools in bench-scale wastewater bioreactors to monitor HGT rates across a community under various reactor conditions; and (3) characterize the host range of the engineered plasmids and of a suite of environmentally-relevant ARGs under different bioreactor operational conditions. The results of this research will advance the knowledge of the mechanisms that govern HGT of ARGs in wastewater treatment and water reuse systems that will inform management strategies to protect human health.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

打击全球抗菌耐药性全球健康危机的主要挑战是，可以通过称为水平基因转移（HGT）的过程在细菌中共享抗生素耐药基因（ARGS）。 ARGS存在于废水中，对回收废水可能传达的ARG健康影响几乎没有理解。关于导致环境细菌（例如用于治疗临床感染中的废水和致病细菌）的hgt的疾病，几乎不知所动。该项目的目标是破译病原体如何从水和废水系统中存在的环境细菌中获取ARG，并了解如何在水和废水微生物群落中传播ARGS。这项研究将开发生物传感器来监测HGT的ARG，以了解废水处理厂中的简单操作参数如何影响ARG的减少或扩散。如果成功的话，该项目的结果将确定环境中抗生素耐药性转移的方法，并确定在水处理过程中停止这种转移的方法，保护公共卫生和国家的供水。理解对环境中建立的水平基因转移（HGT）的控制能力将赋予抗衰老抗伯伯的抗近亲抗体的抗原抗性能力。尽管有多种技术可用于获得通过HGT获得抗生素耐药基因（ARGS）的细菌的静态快照，但这些现有方法并未提供有关经历不断变化环境的复杂社区中基因流的途径和速率的动态信息。这些方法也无法轻易区分生命和死细菌。将利用两种新兴工具来获取此信息：（1）报告原位共轭事件的气体报告的生物传感器； （2）一种与文化无关的方法，用于确定混合社区中ARG的宿主范围。这些工具将应用于处理家庭废水的生物反应器中，以更好地了解操作控制如何影响ARG的传播率和宿主范围。这项研究的目的是（1）通过耦合一种酶的合成，从而开发出对HGT原位报告的生物传感器，该酶的合成产生了罕见的挥发性气体与宽范围的质粒转移； （2）在台式废水生物反应器中使用这些工具在各种反应堆条件下监测社区的HGT率； （3）在不同的生物反应器操作条件下，工程质粒的宿主范围和与环境相关的ARG的宿主范围。这项研究的结果将促进对废水处理和水再利用系统中ARG的机制的了解，这些机制将为保护人类健康的管理策略提供信息。该奖项反映了NSF的法定任务，并被认为是值得通过基金会的知识分子优点和更广泛影响的审查标准来通过评估来支持的。