Venlafaxine in Embryos Stimulates Neurogenesis and Disrupts Larval Behavior in Zebrafish.

Venlafaxine, a widely prescribed antidepressant, is a selective serotonin and norepinephrine reuptake inhibitor in humans, and this drug is prevalent in municipal wastewater effluents. While studies have shown that this drug affects juvenile fish behavior, little is known about the developmental impact on nontarget aquatic animals. We tested the hypothesis that venlafaxine deposition in the egg, mimicking maternal transfer of this antidepressant, disrupts developmental programming using zebrafish (Danio rerio) as a model. Embryos (1-4 cell stage) were microinjected with either 1 or 10 ng venlafaxine, which led to a rapid reduction (90%) of this drug in the embryo at hatch. There was a concomitant increase in the concentration of the major metabolite o-desmethylvenlafaxine during the same period. Embryonic exposure to venlafaxine accelerated early development, increased hatching rate and produced larger larvae at 5 days post fertilization. Also, there was an increase in neuronal birth in the hypothalamus, dorsal thalamus, posterior tuberculum, and the preoptic region, and this corresponded with a higher spatial expression of nrd4, a key marker of neurogenesis. The venlafaxine-exposed larvae were less active and covered shorter distance in a light and dark behavioral test compared to the controls. Overall, zygotic exposure to venlafaxine disrupts early development, including brain function, and compromises larval behavior, suggesting impact of this drug on developmental programming in zebrafish.

Combining chemical, bioanalytical and predictive tools to assess persistence, seasonality, and sporadic releases of organic micropollutants within the urban water cycle.

Using a novel liquid chromatography-tandem mass spectrometry method with large volume direct injection and quantitation via isotope dilution, we evaluated the presence of 55 organic micropollutants in wastewater effluents, and locations within the Bow River and Elbow River watersheds in and around the city of Calgary, Alberta, Canada. In addition to establishing baseline micropollutant data for water utility operations, our study aimed to enhance our understanding of micropollutant behavior in the urban water cycle, assess the contributions of three wastewater treatment plants (WWTPs) to downstream receiving waters, explain the potential causes of total estrogenicity measured using the yeast-estrogen screen assay (YES), and prioritize a subset of substances for continuous monitoring. With data spanning 48 months and 95 river km, our results indicate the extensive persistence of metformin (antidiabetic), seasonality of N,N­diethyl-m-toluamide (DEET, insect repellant), O-desmethylvenlafaxine (antidepressant metabolite), and sulfamethoxazole (antibiotic) in source waters, and sporadic detections of a well-known perfluoroalkyl substance (PFOA). The seasonality of pharmaceuticals at the sentinel downstream monitoring site appeared to coincide with river dilution while that of DEET was likely attributable to peak usage during the warmer months. Steroidal estrogens were rarely detected in wastewater effluents although total estrogenicity via YES was evident, suggesting the presence of less potent but more abundant non-steroidal estrogens (e.g., flame retardants, bisphenols, and phthalates). A conservative mass balance analysis suggests that the largest WWTP (serving a population of >1 million) consistently contributed the highest load of micropollutants, with the exception of metformin, which appeared to be influenced by a smaller WWTP (serving 115,000) that operates a different activated sludge process. We consider metformin, sucralose, diclofenac, and venlafaxine as more effective conservative tracers of wastewater pollution due to their notably higher concentrations and persistence in the Bow River compared to carbamazepine and caffeine, respectively. Finally, hierarchical clustering revealed a close association between E. coli and caffeine, supporting the use of caffeine as an indicator of short-term, untreated anthropogenic inputs. Overall, this study yields valuable insights on the presence, behavior, and sources of organic micropollutants in the urban water cycle and identifies indicators of anthropogenic impacts that are useful for prioritizing future monitoring campaigns in Calgary and elsewhere.