Metabolomic responses to pre-chlorinated and final effluent wastewater with the addition of a sub-lethal persistent contaminant in Daphnia magna.

Consumer products such as perfluorooctanesulfonic acid (PFOS) and pharmaceuticals (PCPPs) enter aquatic ecosystems through inefficient removal during wastewater treatment. Often, the sterilization process of wastewater includes the addition of sodium hypochlorite that can react with PCPPs and other organic matter (i.e., dissolve organic matter) to generate disinfection by-products and can cause the final effluent to be more harmful to aquatic organisms. Here, we exposed Daphnia magna to two stages of wastewater, the pre-chlorinated wastewater (PreCl) and the final effluent. In addition, we exposed D. magna, to the final effluent with a concentration gradient of added PFOS, to investigate if this persistent contaminant altered the toxicity of the final effluent. After 48 h of contaminant exposure, we measured the daphnids metabolic responses to the different stages of wastewater treatment, and with the addition of PFOS, utilizing proton nuclear magnetic resonance spectroscopy and liquid chromatography tandem mass spectrometry. We found few significant changes to the metabolic profile of animals exposed to the PreCl wastewater; however, animals exposed to the final effluent displayed increases in many amino acids and decreases in some sugar metabolites. With the addition of PFOS to the final effluent, the metabolic profile shifted from increased amino acids and decreased sugar metabolites and energy molecules especially at the low and high concentrations of PFOS. Overall, our results demonstrate the metabolome is sensitive to changes in the final effluent that are caused by sterilization, and with the addition of a persistent contaminant, the metabolic profile is further altered.