Collaborative Research: Impacts of biocides associated with hydraulic fracturing on aquatic microbial communities.

水力压裂（HF），通常被称为“水力压裂”，是一种利用高压水、砂和化学物质破裂岩石，释放石油和天然气的石油和天然气回收方法。这一过程改变了美国的能源工业，但需要更多的研究来研究HF对环境的影响。朱尼亚塔学院与密歇根理工大学和田纳西大学诺克斯维尔分校的研究人员合作，将重点研究杀菌剂对环境的影响，杀菌剂是氟化氢中最常用的一些化学物质。杀菌剂的设计目的是杀死微生物，在HF中用于保护设备免受微生物腐蚀，并保持石油和天然气的质量。然而，一旦接触到杀菌剂，自然产生的微生物就会对这些化学物质产生抗药性。由于杀菌剂耐药性与微生物对药物产生耐药性的能力之间存在这种联系，因此澄清与HF相关的工业杀菌剂对环境的影响以及导致杀菌剂耐药性的过程非常重要。如果取得成功，这项研究将产生更好的战略，以确定和减轻杀菌剂对环境和公众健康的潜在影响，保护国家的水安全，同时使使用一种重要的原始能源材料成为可能。本研究的目的是更好地了解氟化氢对环境的影响，以及在氟化氢操作中使用工业杀菌剂如何促进水生微生物群落的抗微生物药物耐药性（AMR）的发展。先前的研究表明，受HF操作影响的河流中微生物群落组成发生了变化，并且受HF影响的河流对杀菌剂的耐受性增加。本研究选择了宾夕法尼亚州的几条河流，基于它们接收由于泄漏或靠近活性HF井而产生的HF废水。另一组没有主动HF操作的流被选择作为控制设置。这项工作的最初目标将是研究观察到的高HF活性流的变化与HF废水的潜在释放之间的联系。化学示踪剂、同位素特征、杀菌剂浓度和杀菌剂分解产物将在HF影响的溪流中进行测量，并与HF废水和对照溪流进行比较。我们还将利用高通量测序技术研究这3种水生生态系统中的微生物群落组成和功能，以阐明HF操作对生物地球化学循环的影响，并评估微生物作为HF影响敏感生物指标的潜力。除了调查HF操作对河流的影响和AMR的后果外，还将进行研究，利用从HF废水和受HF影响的河流中分离出的抗杀菌剂菌株，研究对各种工业杀菌剂的生物抗性机制。转录组学将用于评估微生物对工业杀菌剂的反应。将在这些环境中确定多年的抗生物杀灭剂抗性菌株和抗菌素耐药性基因水平，以调查HF活性对抗微生物药物抗性菌株的发展和持续存在的长期影响。该项目将深入了解HF活性、杀菌剂耐药性以及在这些环境中控制杀菌剂命运的途径之间的联系。如果成功的话，这项研究可以作为未来检测杀菌剂污染的分子诊断工具开发的基础。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Hydraulic fracturing (HF), commonly called "fracking", is a method for recovery of oil and gas using high pressure water, sand, and chemicals to fracture rocks, releasing oil and gas. This process transformed the U.S. energy industry, but more research is needed to study the environmental impacts of HF. This project at Juniata College, in collaboration with researchers at Michigan Technological University and the University of Tennessee Knoxville, will focus on the environmental impacts of biocides, some of the most commonly used chemicals in HF. Biocides are designed to kill microbes and are used in HF to protect equipment from microbial corrosion and to preserve the quality of oil and gas. However, once exposed to biocides, naturally-occurring microorganisms can become resistant to these chemicals. Because of this link between biocide resistance and the ability of microbes to become resistant to medications, it is important to clarify the impact of industrial biocides associated with HF on the environment, and the processes leading to biocide resistance. If successful, this research will lead to better strategies to identify and mitigate the potential effects of biocides on the environment and public health, protecting the Nation's water security while enabling the use of an important source of raw energy materials. The goal of this study is to better understand the environmental impacts of HF and how the use of industrial biocides in HF operations may contribute to development of antimicrobial resistance (AMR) in aquatic microbial communities. Previous work demonstrated altered microbial community compositions in streams impacted by HF operations as well as increased tolerance to biocides in streams impacted by HF. Several streams in Pennsylvania were selected for this study based on their reception of HF wastewater due to spills or close proximity to active HF wells. Another set of streams with no active HF operations were selected as control settings. The initial objective of this work will be to study links between the observed changes in high HF active streams and potential releases of HF wastewater. Chemical tracers, isotopic signatures, biocide concentrations, and biocide breakdown products will be measured in HF-impacted streams and compared to both HF wastewater and control streams. The microbial community composition and functions within these 3 general aquatic ecosystems will also be studied using high-throughput sequencing to elucidate the impact of HF operations on biogeochemical cycling and assess the potential for microbes to be used as sensitive bio-indicators of HF impacts. In addition to investigating the impact of HF operations on streams and the consequences of AMR, studies will be performed to investigate the biological mechanism for resistance to various industrial biocides using biocide resistant strains isolated from HF wastewater and HF-impacted streams. Transcriptomics will be employed to assess the microbial response to industrial biocides. The levels of biocide-resistant strains and AMR genes will be determined in these environments over multiple years to investigate the long-term impact of HF activities on the development and persistence of strains resistant to antimicrobials. This project will provide insights into links between HF activity, biocide resistance, and the pathways controlling the fate of biocides in these settings. If successful, this research could serve as the foundation for the development of future molecular diagnostic tools for detecting biocide contamination.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.