A pipeline carrying unconventional oil and gas (OG) wastewater spilled approximately 11 million liters of wastewater into Blacktail Creek, North Dakota, USA. Flow of the mix of stream water and wastewater down the channel resulted in storage of contaminants in the hyporheic zone and along the banks, providing a long-term source of wastewater constituents to the stream. A multi-level investigation was used to assess the potential effects of oil and brine spills on aquatic life. In this study, we used a combination of experiments using a native fish species, Fathead Minnow (Pimephales promelas), field sampling of the microbial community structure, and measures of estrogenicity. The fish investigation included in situ experiments and experiments with collected site water. Estrogenicity was measured in collected site water samples, and microbial community analyses were conducted on collected sediments. During the initial post-spill investigation, February 2015, performing in situ fish bioassays was impossible because of ice conditions. However, microbial community (e.g., the presence of members of the Halomonadaceae, a family that is indicative of elevated salinity) and estrogenicity differences were compared to reference sites and point to early biological effects of the spill. We noted water column effects on in situ fish survival 6 months post-spill during June 2015. At that time, total dissolved ammonium (sum of ammonium and ammonia, TAN) was 4.41 mg NH₄/L with an associated NH₃ of 1.09 mg/L, a concentration greater than the water quality criteria established to protect aquatic life. Biological measurements in the sediment defined early and long-lasting effects of the spill on aquatic resources. The microbial community structure was affected during all sampling events. Therefore, sediment may act as a sink for constituents spilled and as such provide an indication of continued and cumulative effects post-spill. However, lack of later water column effects may reflect pulse hyporheic flow of ammonia from shallow ground water. Combining fish toxicological, microbial community structure and estrogenicity information provides a complete ecological investigation that defines potential influences of contaminants at organismal, population, and community levels. In general, in situ bioassays have implications for the individual survival and changes at the population level, microbial community structure defines potential changes at the community level, and estrogenicity measurements define changes at the individual and molecular level. By understanding effects at these various levels of biological organization, natural resource managers can interpret how a course of action, especially for remediation/restoration, might affect a larger group of organisms in the system. The current work also reviews potential effects of additional constituents defined during chemistry investigations on aquatic resources.

一条输送非常规石油和天然气 (OG) 废水的管道将大约 1100 万升废水泄漏到美国北达科他州的黑尾溪。溪流水和废水的混合物沿着渠道向下流动，导致污染物储存在潜流区和沿河岸，为溪流提供了废水成分的长期来源。多层次调查用于评估石油和盐水泄漏对水生生物的潜在影响。在这项研究中，我们使用了本地鱼类 Fathead Minnow ( Pimephales promelas)、微生物群落结构的现场采样和雌激素性测量。鱼类调查包括原位实验和收集现场水的实验。在收集的现场水样中测量了雌激素性，并对收集的沉积物进行了微生物群落分析。在 2015 年 2 月的初步泄漏后调查期间，由于结冰条件，无法进行原位鱼类生物测定。然而，微生物群落（例如，盐单胞菌科成员的存在，表明盐度升高的一个家族）和雌激素差异与参考地点进行了比较，并指出了泄漏的早期生物学效应。我们注意到 2015 年 6 月泄漏后 6 个月水柱对原地鱼类生存的影响。当时，总溶解铵（铵和氨的总和，₄ /L 与相关的 NH ₃ 1.09 毫克/升，浓度高于为保护水生生物而制定的水质标准。沉积物中的生物测量确定了泄漏对水生资源的早期和长期影响。在所有采样事件中，微生物群落结构都受到影响。因此，沉积物可能充当泄漏成分的汇，因此提供了泄漏后持续和累积影响的指示。然而，后期水柱效应的缺乏可能反映了来自浅层地下水的氨脉动潜流流动。结合鱼类毒理学、微生物群落结构和雌激素性信息，可以进行完整的生态调查，确定污染物在生物体、种群和群落层面的潜在影响。一般来说，原位生物测定对个体生存和种群水平的变化有影响，微生物群落结构定义了社区水平的潜在变化，雌激素测量定义了个体和分子水平的变化。通过了解这些不同级别的生物组织的影响，自然资源管理者可以解释一个行动过程，尤其是补救/恢复行动，可能如何影响系统中更大的生物群。目前的工作还审查了在化学调查期间定义的其他成分对水生资源的潜在影响。通过了解这些不同层次的生物组织的影响，自然资源管理者可以解释一个行动过程，尤其是修复/恢复，可能如何影响系统中更大的生物群。目前的工作还审查了在化学调查期间定义的其他成分对水生资源的潜在影响。通过了解这些不同级别的生物组织的影响，自然资源管理者可以解释一个行动过程，尤其是补救/恢复行动，可能如何影响系统中更大的生物群。目前的工作还审查了在化学调查期间定义的其他成分对水生资源的潜在影响。