Using Transcriptomic Tools to Evaluate Biological Effects Across Effluent Gradients at a Diverse Set of Study Sites in Minnesota, USA

The aim of this study was to explore the utility of “omics” approaches in monitoring aquatic environments where complex, often unknown stressors make chemical-specific risk assessment untenable. We examined changes in the fathead minnow (<i>Pimephales promelas</i>) ovarian transcriptome following 4-day exposures conducted at three sites in Minnesota (MN, USA). Within each site, fish were exposed to water from three locations along a spatial gradient relative to a wastewater treatment plant (WWTP) discharge. After exposure, site-specific impacts on gene expression in ovaries were assessed. Using an intragradient point of comparison, biological responses specifically associated with the WWTP effluent were identified using functional enrichment analyses. Fish exposed to water from locations downstream of the effluent discharges exhibited many transcriptomic responses in common with those exposed to the effluent, indicating that effects of the discharge do not fully dissipate downstream. Functional analyses showed a range of biological pathways impacted through effluent exposure at all three sites. Several of those impacted pathways at each site could be linked to potential adverse reproductive outcomes associated with the hypothalamic–pituitary–gonadal (HPG) axis in female fathead minnows, specifically signaling pathways associated with oocyte meiosis, TGF-beta signaling, gonadotropin-releasing hormone (GnRH) and epidermal growth factor receptor family (ErbB), and gene sets associated with cyclin B-1 and metalloproteinase. The utility of this approach comes from the ability to identify biological responses to pollutant exposure, particularly those that can be tied to adverse outcomes at the population level and those that identify molecular targets for future studies