Multiple Stressors in the Environment: The Effects of Exposure to an Antidepressant (Venlafaxine) and Increased Temperature on Zebrafish Metabolism

Aquatic organisms are continuously exposed to multiple environmental stressors working cumulatively to alter ecosystems. Wastewater-dominated environments are often riddled by a myriad of stressors, such as chemical and thermal stressors. The objective of this study was to examine the effects of an environmentally relevant concentration of a commonly prescribed antidepressant, venlafaxine (VFX) [1.0 μg/L], in addition to a 5°C increase in water temperature on zebrafish metabolism. Fish were chronically exposed (21 days) to one of four conditions: (i) 0 μg/L VFX at 27°C; (ii) 1.0 μg/L VFX at 27°C; (iii) 0 μg/L VFX at 32°C; (iv) 1.0 μg/L VFX at 32°C. Following exposure, whole-body metabolism was assessed by routine metabolic rate (RMR) measurements, whereas tissue-specific metabolism was assessed by measuring the activities of major metabolic enzymes in addition to glucose levels in muscle. RMR was significantly higher in the multi-stressed group relative to Control. The combination of both stressors resulted in elevated pyruvate kinase activity and glucose levels, while lipid metabolism was depressed, as measured by 3-hydroxyacyl CoA dehydrogenase activity. Citrate synthase activity increased with the onset of temperature, but only in the group treatment without VFX. Catalase activity was also elevated with the onset of the temperature stressor, however, that was not the case for the multi-stressed group, potentially indicating a deleterious effect of VFX on the anti-oxidant defense mechanism. The results of this study highlight the importance of multiple-stressor research, as it able to further bridge the gap between field and laboratory studies, as well as have the potential of yielding surprising results that may have not been predicted using a conventional single-stressor approach.This study was supported by the Natural Sciences and Engineering Research Council of Canada (NSERC: RGPIN-2015-05643) as well as the Canada Foundation for Innovation (CFI 34317)

Impacts of wastewater treatment plant effluent on energetics and stress response of rainbow darter (Etheostoma caeruleum) in the Grand River watershed

The final publication is available at Elsevier via https://doi.org/10.1016/j.cbpb.2017.11.011. © 2018. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/The objective of this study was to assess the effects of municipal wastewater treatment plant effluent on the energetics and stress response of rainbow darter (Etheostoma caeruleum). Male and female rainbow darter were collected upstream and downstream of the Waterloo WWTP in the Grand River watershed, ON, Canada. To assess the effects of wastewater treatment plant effluent on whole-body and tissue specific metabolic capacity, closed-chamber respirometry and muscle-enzyme activity analyses were performed. Plasma cortisol was also collected from fish before and after an acute air-exposure stressor to evaluate the cortisol stress response in fish exposed to additional stressors. Male and female rainbow darter collected downstream of the effluent had higher oxygen consumption rates, while differences in enzyme activities were primarily associated with sex rather than collection site. No impairment in the cortisol stress response between downstream and upstream fish was observed, however baseline cortisol levels in female fish from the downstream site were significantly higher compared to other baseline groups. Stress-induced cortisol levels were also higher in female fish from both sites when compared to their male counterparts. Overall, this study demonstrates that chronic exposure to WWTP effluent impacts whole-body metabolic performance. This study was also able to demonstrate that sex-differences are a key determinant of various metabolic changes in response to physiological stress, thereby, providing a novel avenue to be considered and further explored.Natural Sciences and Engineering Research Council (Grant RGPIN-2015-05643)Canada Foundation for Innovation (Grant CFI 34317

Impacts on Metabolism and Gill Physiology of Darter Species (Etheostoma spp.) That Are Attributed to Wastewater Effluent in the Grand River

The effluent from municipal wastewater treatment plants is a major point source of contamination in Canadian waterways. The improvement of effluent quality to reduce contaminants, such as pharmaceuticals and personal care products, before being released into the environment is necessary to reduce the impacts on organisms that live in the river downstream. Here, we aimed to characterize the metabolic and gill physiological responses of rainbow (Etheostoma caeruleum), fantail (Etheostoma flabellare), and greenside (Etheostoma blennioides) darters to the effluent in the Grand River from the recently upgraded Waterloo municipal wastewater treatment plant. The routine metabolism of darters was not affected by effluent exposure, but some species had increased maximum metabolic rates, leading to an increased aerobic scope. The rainbow darter aerobic scope increased by 2.2 times and the fantail darter aerobic scope increased by 2.7 times compared to the reference site. Gill samples from effluent-exposed rainbow darters and greenside darters showed evidence of more pathologies and variations in morphology. These results suggest that darters can metabolically adjust to effluent-contaminated water and may also be adapting to the urban and agricultural inputs. The modification and damage to the gills provide a useful water quality indicator but does not necessarily reflect how well acclimated the species is to the environment due to a lack of evidence of poor fish health.This research was supported through the Natural Science and Engineering Research Council (#RGPIN-2015-05643) and the Canadian First Research Excellence Fund Global Water Futures initiative (PI: Mark Servos). R.H. was supported by a Queen Elizabeth II Graduate Scholarship in Science and Technology

Tissue storage affects lipidome profiling in comparison to in vivo microsampling approach

Low-invasive in vivo solid-phase microextraction (SPME) was used to investigate the lipid profiles of muscle tissue of living fish. Briefly, mixed mode SPME fibers were inserted into the muscle for 20 min extraction, and then the fibers were desorbed in an optimal mixture of solvents. The obtained lipid profile was then compared and contrasted to that obtained with employment of ex vivo SPME and solid-liquid extraction (SLE) from fish muscle tissue belonging to the same group of fish, following a one-year storage period. Ex vivo SPME analysis of stored muscle samples revealed 10-fold decrease in the number of detected molecular features in comparison to in vivo study. Moreover, in vivo microsampling enabled the identification of different classes of bioactive lipids, including fatty acyls, not present in the lipid profile obtained through ex vivo SPME and SLE, suggesting the alterations occurring in the unbound lipid fraction of the system under study during the storage and also indicating the advantage of the in vivo extraction approach.This work was supported by Environment Canada through the Environmental Damages Fund (Grant EC-129114)

In vivo microsampling to capture the elusive exposome

Bessonneau, V., Ings, J., McMaster, M., Smith, R., Bragg, L., Servos, M., & Pawliszyn, J. (2017). In vivo microsampling to capture the elusive exposome. Scientific Reports, 7, 44038. The final and definitive publication is available through Nature publishing Group via: http://dx.doi.org/10.1038/srep44038Loss and/or degradation of small molecules during sampling, sample transportation and storage can adversely impact biological interpretation of metabolomics data. In this study, we performed in vivo sampling using solid-phase microextraction (SPME) in combination with non-targeted liquid chromatography and high-resolution tandem mass spectrometry (LC-MS/MS) to capture the fish tissue exposome using molecular networking analysis, and the results were contrasted with molecular differences obtained with ex vivo SPME sampling. Based on 494 MS/MS spectra comparisons, we demonstrated that in vivo SPME sampling provided better extraction and stabilization of highly reactive molecules, such as 1-oleoyl-sn-glycero-3-phosphocholine and 1-palmitoleoyl-glycero-3-phosphocholine, from fish tissue samples. This sampling approach, that minimizes sample handling and preparation, offers the opportunity to perform longitudinal monitoring of the exposome in biological systems and improve the reliability of exposure-measurement in exposome-wide association studies.Environment Canada, Environmental Damages Fund (Grant EC-129114) provided to Environment Canada through the Joint Oil Sands Monitoring Program

Effect of Background Water Matrices on Pharmaceutical and Personal Care Product Removal by UV-LED/TiO2

In this study, we evaluated the effectiveness of UV-LED-irradiated TiO2 in removing 24 commonly detected PPCPs in two water matrices (municipal wastewater effluent and Suwannee River NOM–synthetic water) and compared their performance with that of ultrapure water. Relatively fast removal kinetics were observed for 29% and 12% of the PPCPs in ultrapure water and synthetic surface water, respectively (kapp of 1–2 min−1). However, they all remained recalcitrant to photocatalysis when using wastewater effluent as the background matrix (kapp < 0.1 min−1). We also observed that the pH-corrected octanol/water partition coefficient (log Dow) correlated well with PPCP degradation rate constants in ultrapure water, whereas molecular weight was strongly associated with the rate constants in both synthetic surface water and wastewater. The electrical energy per order (EEO) values calculated at the end of the experiments suggest that UV-LED/P25 can be an energy-efficient method for water treatment applications (2.96, 4.77, and 16.36 kW h m−3 in ultrapure water, synthetic surface water, and wastewater effluents, respectively). Although TiO2 photocatalysis is a promising approach in removing PPCPs, our results indicate that additional challenges need to be overcome for PPCPs in more complex water matrices, including an assessment of photocatalytic removal under different background water matrices.Canada Research Chair||Natural Sciences and Engineering Research Council||Strategic Project Grant (STPGP 430654-12)||Schwartz-Resiman Foundatio

Utilizing UV-LED pulse width modulation on TiO2 advanced oxidation processes to enhance the decomposition efficiency of pharmaceutical micropollutants

The final publication is available at Elsevier via https://doi.org/10.1016/j.cej.2018.12.065. © 2018 This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/The presence of pharmaceutical and personal care products (PPCPs) in aquatic systems has been a growing cause for concern. Advanced oxidation processes such as UV/TiO2 (ultraviolet light/titanium dioxide) can break down PPCPs into smaller constituents, reducing the pharmaceutical activity. However, this process is limited by low photonic efficiency under UV systems. Controlled periodic illumination (CPI) is a promising solution to overcome the issues concerning low photonic efficiencies. Using a CPI controlled UV-LED/TiO2 process, a mixture of eighteen PPCP compounds were analyzed for their degradation removal on porous titanium – titanium dioxide (PTT) substrates. The kinetic rate constants of PPCPs may be analyzed using multiple regression analysis with parameters such as net charge at experimental pH, solubility, and molecular weight. Negatively charged PPCP compounds were found to have the highest removal compared to neutral and positively charged compounds due to electrostatic attraction forces. Decreasing the duty cycle under CPI or the UV-LED illumination period did not significantly change the individual and cumulative PPCP compound removal, suggesting that the CPI controlled UV-LED/TiO2 processes using PTT substrates were effective in reducing energy requirements without sacrificing removal performance.Natural Sciences and Engineering Research Council [STPGP430654-12]Schwartz-Resiman FoundationWaterloo-Technion Research Co-operation Progra

Author Correction: c-Rel orchestrates energy-dependent epithelial and macrophage reprogramming in fibrosis

Correction to: Nature Metabolism https://doi.org/10.1038/s42255-020-00306-2, published online 9 November 2020. In the version of this article initially published, in the ×40 diseased human kidney images in Supplementary Fig. 1, the FSGS image duplicated the DN image. The error has been corrected in the HTML version of the article

c-Rel orchestrates energy-dependent epithelial and macrophage reprogramming in fibrosis

Fibrosis is a common pathological feature of chronic disease. Deletion of the NF-κB subunit c-Rel limits fibrosis in multiple organs, although the mechanistic nature of this protection is unresolved. Using cell-specific gene-targeting manipulations in mice undergoing liver damage, we elucidate a critical role for c-Rel in controlling metabolic changes required for inflammatory and fibrogenic activities of hepatocytes and macrophages and identify Pfkfb3 as the key downstream metabolic mediator of this response. Independent deletions of Rel in hepatocytes or macrophages suppressed liver fibrosis induced by carbon tetrachloride, while combined deletion had an additive anti-fibrogenic effect. In transforming growth factor-β1-induced hepatocytes, c-Rel regulates expression of a pro-fibrogenic secretome comprising inflammatory molecules and connective tissue growth factor, the latter promoting collagen secretion from HMs. Macrophages lacking c-Rel fail to polarize to M1 or M2 states, explaining reduced fibrosis in RelΔLysM mice. Pharmacological inhibition of c-Rel attenuated multi-organ fibrosis in both murine and human fibrosis. In conclusion, activation of c-Rel/Pfkfb3 in damaged tissue instigates a paracrine signalling network among epithelial, myeloid and mesenchymal cells to stimulate fibrogenesis. Targeting the c-Rel–Pfkfb3 axis has potential for therapeutic applications in fibrotic disease