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Assessing Performance of Wastewater Treatment Using in Vitro Cell-based Assays

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Abstract

Bioanalytical tools, namely in vitro bioassays, can be employed in tandem with chemical analyses to assess the efficacy of wastewater treatment and the potential for adverse effects from the discharges of wastewater into receiving waters. In the present study, samples of untreated wastewater (i.e., influent) and treated wastewater (i.e., effluent) were collected from two wastewater treatment plants and a wastewater treatment lagoon to investigate potential differences in treatment performance. In addition, grab samples of surface water were collected downstream of the lagoon discharge to evaluate the water quality in the receiving stream. After solid-phase extraction (SPE) using ion exchange columns for basic/neutral and acidic compounds, respectively, the extracts were analyzed for a suite of 16 indicator compounds. The two SPE extracts were combined for analysis of biological responses in four in vitro cell-based bioassays. The concentrations of several indicator compounds, including the estrogens, 17β-estradiol (E2) and 17α-ethinylestradiol (EE2), were below the limits of detection. However, androstenedione and estrone were detected in several influent samples. The concentrations of these steroid hormones and some of the other indicator compounds declined during treatment, but acesulfame K, carbamazepine, trimethoprim and DEET persisted in the effluent. The MTS-CellTiter 96® AQueous One Solution Cell Proliferation Assay (MTS) indicated that cell viability was not affected by exposure to the extracts. The Qiagen Nuclear Receptors 10-Pathway Reporter Array indicated that several cellular pathways were upregulated, with the greatest upregulation observed with the estrogen receptor (i.e., induction ratios of 12 to 47) and the liver X receptor (i.e., induction ratios of 10 to 45). The ERα CALUX assay indicated that estrogenic activity was lower in effluents compared to influents, but the expected improved removal of estrogenic activity during nitrification was not observed. The results of the Nrf2 Luciferase Luminescence Assay indicated a lower oxidative stress in the effluent samples, except for the lagoon. Overall, the present study further demonstrates that bioassays provide complementary information to chemical analyses and offer a way to assess treatment performance, even when target contaminants are not detected. There are thus advantages to using a combination of chemical analyses and in vitro bioassays to monitor the treatment efficiency of wastewater treatment plants and to predict the potential impacts of wastewater discharges into receiving waters.

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Acknowledgements

This research was funded through the Canada–Ontario Agreement on Great Lakes Water Quality and Ecosystem Health. Such support does not indicate endorsement by the Government of Ontario of the contents of this contribution. For their assistance in obtaining samples, the authors would like to thank Shirley Anne Smyth, Quintin Rochfort, Scott Alexander and Steven Teslic from Environment and Climate Change Canada, and all staff at the wastewater treatment plants who made the sampling possible. We also thank Rachel Benoit, Marco Pineda and Linda Taylor for the preparation and chemical analysis of the samples as well as Peter Behnisch and Emiel Felzel at Biodetection Systems (Amsterdam) for their guidance and helpful insights in running the ERα CALUX assay.

Funding

The research was funded through the Canada–Ontario Agreement on Great Lakes Water Quality and Ecosystem Health.

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Authors and Affiliations

  1. Department of Chemical Engineering, McGill University, 3610 University St., Montreal, QC, H3A 0C5, Canada

    Adamo Riccardo Petosa & Viviane Yargeau

  2. Ontario Ministry of the Environment, Conservation and Parks, Toronto, ON, Canada

    Monica Nowierski

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  1. Adamo Riccardo Petosa
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  2. Monica Nowierski
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Correspondence to Viviane Yargeau.

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Petosa, A.R., Nowierski, M. & Yargeau, V. Assessing Performance of Wastewater Treatment Using in Vitro Cell-based Assays. Arch Environ Contam Toxicol 82, 21–36 (2022). https://doi.org/10.1007/s00244-021-00900-9

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