Transcriptome signatures of wastewater effluent exposure in larval zebrafish vary with seasonal mixture composition in an effluent-dominated stream

Authors

Emma B. Meade, University of Wisconsin-Milwaukee
Luke R. Iwanowicz, U.S. Geological Survey
Nicklaus Neureuther, University of Wisconsin-Milwaukee
Gregory H. LeFevre, University of Iowa, IIHR-Hydroscience & Engineering
Dana W. Kolpin, U.S. Geological Survey
Hui Zhi, University of Iowa, IIHR-Hydroscience & Engineering
Shannon M. Meppelink, U.S. Geological Survey
Rachael F. Lane, U.S. Geological Survey
Angela Schmoldt, University of Wisconsin-Milwaukee
Aurash Mohaimani, University of Wisconsin-Milwaukee
Olaf Mueller, University of Wisconsin-Milwaukee
Rebecca D. Klaper, University of Wisconsin-Milwaukee

Date of this Version

9-23-2022

Citation

Science of the Total Environment 856 (2023) 159069. http://dx.doi.org/10.1016/j.scitotenv.2022.159069

Comments

U.S. government works are not subject to copyright.

Abstract

Wastewater treatment plant (WWTP) effluent-dominated streams provide critical habitat for aquatic and terrestrial organisms but also continually expose them to complex mixtures of pharmaceuticals that can potentially impair growth, behavior, and reproduction. Currently, few biomarkers are available that relate to pharmaceutical-specific mechanisms of action. In the experiment reported in this paper, zebrafish (Danio rerio) embryos at two developmental stages were exposed to water samples from three sampling sites (0.1 km upstream of the outfall, at the effluent outfall, and 0.1 km below the outfall) during base-flow conditions from two months (January and May) of a temperate-region effluent-dominated stream containing a complex mixture of pharmaceuticals and other contaminants of emerging concern. RNA-sequencing identified potential biological impacts and biomarkers of WWTP effluent exposure that extend past traditional markers of endocrine disruption. Transcriptomics revealed changes to a wide range of biological functions and pathways including cardiac, neurological, visual, metabolic, and signaling pathways. These transcriptomic changes varied by developmental stage and displayed sensitivity to variable chemical composition and concentration of effluent, thus indicating a need for stage-specific biomarkers. Some transcripts are known to be associated with genes related to pharmaceuticals that were present in the collected samples. Although traditional biomarkers of endocrine disruption were not enriched in either month, a high estrogenicity signal was detected upstream in May and implicates the presence of unidentified chemical inputs not captured by the targeted chemical analysis. This work reveals associations between bioeffects of exposure, stage of development, and the composition of chemical mixtures in effluent-dominated surface water. The work underscores the importance of measuring effects beyond the endocrine system when assessing the impact of bioactive chemicals in WWTP effluent and identifies a need for non-targeted chemical analysis when bioeffects are not explained by the targeted analysis.