Preliminary values and standard operating procedures for determining fecal pollution sources in Utah watersheds

We collected water samples from the Logan River, Red Butte Creek, and Provo River in Utah in June 2016 to assess the amount of human, ruminant, and dog fecal contamination in these watersheds. Samples were collected in sterile 1L bottles at sites where the iUTAH NSF EPSCoR program has installed aquatic sensor stations along a mountain-to-urban gradient in each watershed. After collection, we filtered samples through 0.2um PES filters, extracted the sample DNA using a MoBio PowerSoil DNA extraction kit, and analyzed the samples via qPCR.

By identifying the major sources of fecal pollution along urbanization gradients in Utah, we hope to inform efforts to manage fecal pollution in Utah's watersheds. In addition to creating generating raw qPCR values that show the relative contributions of each type (human, ruminant, and dog) to fecal pollution in the rivers sampled, we selected primer sets that we hope will be adopted as standard in future fecal pollution assessment efforts. We also created a standard operating procedure for the creation of qPCR standards, which will allow future researchers to determine total numbers of fecal indicator bacteria present in water samples.

Funded by the iUTAH NSF EPSCoR program (National Science Foundation, iUTAH-innovative Urban Transitions and Aridregion Hydro-sustainability, NSF Award Number 1208732) and conducted at Brigham Young University.

于2016年6月，我们对犹他州的洛根河、红岩溪和普罗沃河进行了水样采集，以评估这些流域中人类、反刍动物和犬类粪便污染的程度。水样于无菌1升瓶中收集，采集地点位于每个流域内，iUTAH NSF EPSCoR项目（国家科学基金会，iUTAH-创新城市转型与干旱地区水可持续性，国家科学基金会资助编号1208732）沿山脉至城市梯度安装了水生传感器站。采集后，我们通过0.2微米的PES滤膜过滤样本，利用MoBio PowerSoil DNA提取试剂盒提取样本DNA，并通过实时定量PCR（qPCR）技术进行分析。通过识别犹他州城市化梯度中粪便污染的主要来源，我们旨在为管理犹他州流域的粪便污染提供信息。此外，我们创建了展示每种类型（人类、反刍动物和犬类）对河流样本中粪便污染相对贡献的原始qPCR值，并选择了有望在未来粪便污染评估工作中被采纳为标准的引物组合。我们还制定了一套标准操作程序，用于qPCR标准的创建，这将允许未来的研究人员确定水样中存在的粪便指示菌的总数。该项目由iUTAH NSF EPSCoR项目资助，并在杨百翰大学执行。