Morphological and behavioral alterations in zebrafish larvae after exposure to contaminated river sediments collected in different weather conditions

Wastewater treatment plants (WWTPs) are the primary source of micropollutants in aquatic ecosystems. Many micropollutants tend to bind to sediments and persist until remobilizion by bioturbation or flood events. Advanced ef-fluent treatment by ozonation has been proven to eliminate most micropollutants.The present study characterizes sediments' toxic potential regarding zebrafish embryo development, which highly complex nervous system is vulnerable to exposure to neurotoxic substances. Furthermore, behavioral changes can be induced even at low pollutant concentrations and do not cause acute toxicity.The study area includes stretches of the main waterbody, the Wurm River (sampling sites W1-W5), and its tributary the Haarbach River (sampling sites H1, and H2) in North-Rhine Westphalia, Germany. Both waterbodies serve as recipi-ents of WWTPs' effluents. The effluent entering the Haarbach River is conventionally treated, while the Wurm River receives ozonated effluent from the Aachen-Soers WWTP. Seven sampling sites up-and downstream of the WWTPs were investigated in June of two subsequent years. The first sampling campaign in 2017 was characterized by pro-longed dry weather. The second sampling campaign in 2018 occurred after prolonged rain events and the release of the rainwater overflow basin.Direct exposure of zebrafish embryos to native sediments using the sediment contact test represented an ecologically realistic scenario and showed no acute sublethal effects. Exposure of the zebrafish embryo to freeze-dried sediments representing the ecotoxicological status of sediments during flood events unfolded acute sublethal toxicity. Behavioral studies with zebrafish larvae were an essential part of environmental neurotoxicity testing. Zebrafish larvae exposed to sediments' concentrations causing no acute effects led to behavioral changes signalizing neurotoxic substances in sed-iments. Polyaromatic hydrocarbons, polychlorinated biphenyls, and nitroaromatic compounds were identified as po-tential toxicity drivers, whereby the rainwater overflow basin served as a possible source of pollution. Mixture toxicity, effect-directed analysis, and further sediment monitoring are needed.

Automated summary

Wastewater treatment plants are the primary source of micropollutants in aquatic ecosystems. Advanced effluent treatment using ozonation has been proven effective in eliminating most micropollutants. This study focuses on the toxic potential of sediments in relation to zebrafish embryo development and behavior. Sublethal effects and neurotoxic substances were observed even at low pollutant concentrations. Polyaromatic hydrocarbons, polychlorinated biphenyls, and nitroaromatic compounds were identified as potential drivers of toxicity.