journal article·2025-07-11Open access

Is Ozonation Treatment Efficient to Provide Safe Reclaimed Water? Assessing the Effects of Synthetic Wastewater Effluents in Human Cell Models

Ana Teresa Rocha, Fátima Jesus, Helena Oliveira, João Gomes, Joana Luísa Pereira

Applied Sciences· Volume 15· Issue 14

Abstract

Ozonation has been promoted as a successful methodology for recovering effluents from wastewater treatment plants, with special emphasis on wastewater contaminated with pharmaceutical and personal care products (PPCPs). Still, ozonation reactions may generate potentially toxic by-products, jeopardizing human health safety, a critical aspect considering the use of reclaimed water. We aimed at understanding the potential impacts of ozonation on the quality of reclaimed water for human use through cell viability assays with human skin keratinocytes (HaCaT cell line). Under this context, the cytotoxicity of synthetic effluents contaminated with methyl- and propylparaben, paracetamol, sulfamethoxazole, and carbamazepine, both individually and in mixtures, was assessed before and after ozonation. The viability of HaCaT cells decreased after exposure to untreated synthetic effluents, denoting the cytotoxicity of the tested PPCPs singly and more prominently in mixtures (especially in those combining two and three PPCPs). A similar pattern was observed when testing effluents treated with ozonation. Since the parent contaminants were fully removed during ozonation, the observed cytotoxicity relates to degradation by-products and interactive effects among them. This study suggests that ozonation is poorly efficient in reducing cytotoxicity, as required for the safe use of ozone-treated reclaimed water in activities involving direct contact with human skin.
Source: crossref · Quality: baseline

Research context

OpenAlex topics: Water Treatment and Disinfection · Wastewater Treatment and Reuse · Water Quality Monitoring Technologies

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

Affiliations

Department of Biology, University of Aveiro, 3810-193 Aveiro, Portugal

CESAM—Centre for Environmental and Marine Studies, University of Aveiro, 3810-193 Aveiro, Portugal

CERES, Department of Chemical Engineering, Faculty of Sciences and Technology, University of Coimbra, 3030-790 Coimbra, Portugal

Funding

CESAM · CERES · Scientific Employment Stimulus

References (47)

Show source-provided references
1. Santos, A.F., Alvarenga, P., Gando-Ferreira, L.M., and Quina, M.J. (2023). Urban Wastewater as a Source of Reclaimed Water for Irrigation: Barriers and Future Possibilities. Environments, 10.10.3390/environments10020017 ↗
2. Wastewater Reclamation and Reuse Trends in Turkey: Opportunities and Challenges10.1016/j.jwpe.2017.10.001 ↗
3. Garrido-Cardenas, J.A., Esteban-García, B., Agüera, A., Sánchez-Pérez, J.A., and Manzano-Agugliaro, F. (2020). Wastewater Treatment by Advanced Oxidation Process and Their Worldwide Research Trends. Int. J. Environ. Res. Public Health, 17.10.3390/ijerph17010170 ↗
4. Modeling the Fate and Human Health Impacts of Pharmaceuticals and Personal Care Products in Reclaimed Wastewater Irrigation for Agriculture10.1016/j.envpol.2021.116532 ↗
5. Kumar, S., Talan, A., Boyle, K., Ormeci, B., Drogui, P., and Tyagi, R.D. (2021). Water Recycling: Economic and Environmental Benefits. Biomass, Biofuels, Biochemicals. Circular Bioeconomy—Current Developments and Future Outlook, Elsevier.10.1016/B978-0-12-821878-5.00015-5 ↗
6. The Utilization of Reclaimed Water: Possible Risks Arising from Waterborne Contaminants10.1016/j.envpol.2019.113020 ↗
7. Assessing Hospital Impact on Pharmaceutical Levels in a Rural ‘Source-to-Sink’ Water System10.1016/j.scitotenv.2020.139618 ↗
8. Chemical Characterization of Riverine Sediments Affected by Wastewater Treatment Plant Effluent Discharge10.1016/j.scitotenv.2022.156305 ↗
9. Source-Specific Ecological Risks and Critical Source Identification of PPCPs in Surface Water: Comparing Urban and Rural Areas10.1016/j.scitotenv.2022.158792 ↗
10. Pharmaceuticals and Personal Care Products (PPCPs) in the Freshwater Aquatic Environment10.1016/j.emcon.2016.12.004 ↗
11. Comparative Analysis of Separation Methods Used for the Elimination of Pharmaceuticals and Personal Care Products (PPCPs) from Water—A Critical Review10.1016/j.seppur.2022.120797 ↗
12. Current Status of Microbes Involved in the Degradation of Pharmaceutical and Personal Care Products (PPCPs) Pollutants in the Aquatic Ecosystem10.1016/j.envpol.2022.118922 ↗
13. Adsorption of Pharmaceutical Pollutants from Water Using Covalent Organic Frameworks10.1002/chem.201801649 ↗
14. Catalytic Ozonation for Water and Wastewater Treatment: Recent Advances and Perspective10.1016/j.scitotenv.2019.135249 ↗
15. Application of Ozonation for Pharmaceuticals and Personal Care Products Removal from Water10.1016/j.scitotenv.2017.01.216 ↗
16. Advanced Oxidation Process-Mediated Removal of Pharmaceuticals from Water: A Review10.1016/j.jenvman.2018.04.103 ↗
17. Perceptions on Ozonation Water Treatment Use: An Alternative Idea for Asean Water Resource Sustainability
18. Jesus, F., Bernardo, C., Martins, R.C., Gomes, J., and Pereira, J.L. (2022). Ecotoxicological Consequences of the Abatement of Contaminants of Emerging Concern by Ozonation—Does Mixture Complexity Matter?. Water, 14.10.3390/w14111801 ↗
19. Ecotoxicity Variation through Parabens Degradation by Single and Catalytic Ozonation Using Volcanic Rock10.1016/j.cej.2018.11.194 ↗
20. Photocatalytic Oxidation of Pharmaceutical Contaminants of Emerging Concern Using Sunlight and Visible Radiation: Mechanism and Ecotoxicological Evaluation10.1016/j.jwpe.2021.102204 ↗
21. Assessing Environmental Risk of Pharmaceuticals in Portugal: An Approach for the Selection of the Portuguese Monitoring Stations in Line with Directive 2013/39/EU10.1016/j.chemosphere.2015.10.100 ↗
22. Gomes, J., Bernardo, C., Jesus, F., Pereira, J.L., and Martins, R.C. (2022). Ozone Kinetic Studies Assessment for the PPCPs Abatement: Mixtures Relevance. Chem. Eng., 6.10.3390/chemengineering6020020 ↗
23. Occurrence and removal of pharmaceuticals in wastewater treatment plants10.1016/j.psep.2021.04.045 ↗
24. Saeid, S., Kråkström, M., Tolvanen, P., Kumar, N., Eränen, K., Mikkola, J.-P., Kronberg, L., Eklund, P., Peurla, M., and Aho, A. (2020). Advanced Oxidation Process for Degradation of Carbamazepine from Aqueous Solution: Influence of Metal Modified Microporous, Mesoporous Catalysts on the Ozonation Process. Catalysts, 10.10.3390/catal10010090 ↗
25. Cytotoxicity and Mutagenicity Study of Waste and Purified Water Samples from Electroplating Industries Prepared by Use of Ferrous Sulfate and Wood Fly Ash10.1081/ESE-200056120 ↗
26. Citotoxicity Status of Electroplating Wastewater Prior/after Neutralization/Purification with Alkaline Solid Residue of Electric Arc Furnace Dust10.1080/10934520802597945 ↗
27. The Assessment of Genotoxic Effects of Wastewater from a Fertilizer Factory10.1002/jat.1381 ↗
28. Assessment of Hepatotoxicity and Dermal Toxicity of Butyl Paraben and Methyl Paraben Using HepG2 and HDFn In Vitro Models10.1016/j.tiv.2018.12.007 ↗
29. Mechanism of P-Hydroxybenzoate Ester-Induced Mitochondrial Dysfunction and Cytotoxicity in Isolated Rat Hepatocytes10.1016/S0006-2952(98)00097-5 ↗
30. Apoptotic Effects of Protocatechuic Acid in Human Breast, Lung, Liver, Cervix, and Prostate Cancer Cells: Potential Mechanisms of Action10.1021/jf9004466 ↗
31. Protocatechuic Acid Induces Cell Death in HepG2 Hepatocellular Carcinoma Cells through a C-Jun N-Terminal Kinase-Dependent Mechanism10.1007/s10565-006-0082-4 ↗
32. Kalinowska, M., Gołębiewska, E., Świderski, G., Męczyńska-Wielgosz, S., Lewandowska, H., Pietryczuk, A., Cudowski, A., Astel, A., Świsłocka, R., and Samsonowicz, M. (2021). Plant-derived and dietary hydroxybenzoic acids—A comprehensive study of structural, anti-/pro-oxidant, lipophilic, antimicrobial, and cytotoxic activity in mda-mb-231 and mcf-7 cell lines. Nutrients, 13.10.3390/nu13093107 ↗
33. Effects of Single and Repeated In Vitro Exposure of Three Forms of Parabens, Methyl-, Butyl- and Propylparabens on the Proliferation and Estradiol Secretion in MCF-7 and MCF-10A Cells10.1016/S1734-1140(13)71024-7 ↗
34. Glucocorticoid-like Activity of Propylparaben, Butylparaben, Diethylhexyl Phthalate and Tetramethrin Mixtures Studied in the MDA-Kb2 Cell Line10.1016/j.toxlet.2014.11.019 ↗
35. Propylparaben-Induced Disruption of Energy Metabolism in Human HepG2 Cell Line Leads to Increased Synthesis of Superoxide Anions and Apoptosis10.1016/j.tiv.2015.11.011 ↗
36. Fouyet, S., Olivier, E., Leproux, P., Dutot, M., and Rat, P. (2022). Pregnant Women and Endocrine Disruptors: Role of P2X7 Receptor and Mitochondrial Alterations in Placental Cell Disorders. Cells, 11.10.3390/cells11030495 ↗
37. Ottaviano, E., Baron, G., Fumagalli, L., Leite, J., Colombo, E.A., Artasensi, A., Aldini, G., and Borghi, E. (2021). Candida Albicans Biofilm Inhibition by Two Vaccinium Macrocarpon (Cranberry) Urinary Metabolites: 5-(3′,4′-DihydroxyPhenyl)-γ-Valerolactone and 4-Hydroxybenzoic Acid. Microorganisms, 9.10.3390/microorganisms9071492 ↗
38. Mittraphab, Y., Amen, Y., Nagata, M., Matsumoto, M., Wang, D., and Shimizu, K. (2022). Anti-Phototoxicity Effect of Phenolic Compounds from Acetone Extract of Entada Phaseoloides Leaves via Activation of COX-2 and INOS in Human Epidermal Keratinocytes. Molecules, 27.10.3390/molecules27020440 ↗
39. Antagonist Effects of Veratric Acid against UVB-Induced Cell Damages10.3390/molecules18055405 ↗
40. Acetaminophen Cytotoxicity in HepG2 Cells Is Associated with a Decoupling of Glycolysis from the TCA Cycle, Loss of NADPH Production, and Suppression of Anabolism10.1007/s00204-018-2371-0 ↗
41. The Cytotoxicity of Oxalate, Metabolite of Ethylene Glycol, Is Due to Calcium Oxalate Monohydrate Formation10.1016/j.tox.2004.11.029 ↗
42. Oxalate Is Toxic to Renal Tubular Cells Only at Supraphysiologic Concentrations10.1111/j.1523-1755.2005.00576.x ↗
43. DNA Damage Induced by Hydroquinone Can Be Prevented by Fungal Detoxification10.1016/j.toxrep.2014.10.024 ↗
44. Monteiro, A. (2018). Preparação e Caracterização de Nanofibras Magnéticas Para Liberação Controlada de Fármacos e Hipertemia. [Ph.D Thesis, Universidade Federal de Minas Gerais].
45. Effect of Topical Neuromodulatory Medications on Oral and Skin Keratinocytes10.1111/jop.12437 ↗
46. Chlorination, Chloramination and Ozonation of Carbamazepine Enhance Cytotoxicity and Genotoxicity: Multi-Endpoint Evaluation and Identification of Its Genotoxic Transformation Products10.1016/j.jhazmat.2017.08.076 ↗
47. Study of the influence of the matrix characteristics over the photocatalytic ozonation of parabens using Ag-TiO210.1016/j.scitotenv.2018.07.430 ↗