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| Status |
Public on Aug 01, 2023 |
| Title |
Transcriptome signatures of wastewater effluent exposure in larval zebrafish vary with seasonal mixture composition in an effluent-dominated stream |
| Organism |
Danio rerio |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
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.
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| Overall design |
We investigate associations between gene expression in zebrafish larvae (3 and 6 dpf) and evolving seasonal pharmaceutical mixtures in a wastewater effluent-dominated stream. Embryos were exposed to stream water samples at three sites: 100 meters above the effluent outfall, at the the WWTP effluent outfall, and 100 meters below the effluent outfall. There were 5 replicate exposures per site with 20 individual per replicate for separate 3- and 6-day exposures. Samples used for RNA sequencing met the following quality control parameters: OD260/OD280 ratio 1.8-2.0 and OD260/OD230 ratio 2.0-2.2 and a RIN >7.
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| Contributor(s) |
Meade EB, Iwanowicz LR, Neureuther N, LeFevre GH, Kolpin DW, Zhi H, Meppelink SM, Lane RF, Schmoldt A, Mohaimani A, Mueller O, Klaper RD |
| Citation(s) |
36174698 |
| Submission date |
Jul 02, 2021 |
| Last update date |
Nov 15, 2023 |
| Contact name |
Rebecca Klaper |
| E-mail(s) |
rklaper@uwm.edu
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| Phone |
4143821700
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| Organization name |
University of Wisconsin - Milwaukee
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| Department |
School of Freshwater Sciences
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| Lab |
Klaper
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| Street address |
600 E Greenfield
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| City |
Milwaukee |
| State/province |
WI |
| ZIP/Postal code |
53204 |
| Country |
USA |
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| Platforms (1) |
| GPL14875 |
Illumina HiSeq 2000 (Danio rerio) |
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| Samples (58)
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| Relations |
| BioProject |
PRJNA743294 |
| SRA |
SRP326662 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE179335_RAW.tar |
50.6 Mb |
(http)(custom) |
TAR (of TSV) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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