Contribution of pharmaceuticals, fecal bacteria and endotoxin to the inflammatory responses to inland waters Ahmed El Marghani a,1,2 , Ajay Pradhan a,2 , Asmerom Seyoum a , Hazem Khalaf a , Torbjön Ros b , Lars-Håkan Forsberg c , Tomas Nermark d , Lisa Osterman e , Ulf Wiklund f , Per Ivarsson g , Jana Jass a, ⁎, Per-Erik Olsson a, ⁎⁎ a Örebro Life Science Center, School of Science and Technology, Örebro University, Örebro SE-70182, Sweden b Pelagia Miljökonsult AB, Sjöbod 2, Strömpilsplatsen 12, Umeå 90743, Sweden c Mälarenegi AB, Box 14, Västerås 72103, Sweden d Karlskoga Energi och Miljö AB, Box 42, Karlskoga 69121, Sweden e Skebäcks reningsverk, Box 33300, Örebro 70215, Sweden f Tyréns AB, Västra Norrlandsgatan 10B, 90327 Umeå, Sweden g ALS Scandinavia AB, Box 511, Täby 18325, Sweden HIGHLIGHTS • The present study was performed to determine the distribution of pro-inflammatory effects to inland waters. • Three different wastewater treatment plants and their recipient lakes were studied. • The levels of pharmaceuticals in the WWTPs and lakes were determined and compared to the immune response elicited by urinary bladder and immune cells. • The results of the present study indicate that the cytokine profiles of exposed cells correlate to the endotoxin load of the waters rather than to the levels of pharmaceuticals or live bacteria load. abstract article info Article history: Received 11 December 2013 Received in revised form 22 April 2014 Accepted 22 April 2014 Available online 13 May 2014 Editor: Adrian Covaci Keywords: Wastewater treatment plants Immune response Cytokines Bacteria Inflammation Pharmaceuticals The increasing contamination of freshwater with pharmaceuticals, surfactants, pesticides and other organic compounds are of major concern. As these contaminants are detected at trace levels in the environment it is important to determine if they elicit biological responses at the observed levels. In addition to chemical pollutants, there is also a concern for increasing levels of bacteria and other microorganisms in freshwater systems. In an earlier study, we observed the activation of inflammatory systems downstream of a wastewater treatment plant (WWTP) in southern Sweden. We also observed that the water contained unidentified components that were pro-inflammatory and potentiated the immune response in human urinary bladder epithelial cells. In order to determine if these effects were unique for the studied site or represent a common response in Swedish water, we have now performed a study on three WWTPs and their recipient waters in central Sweden. Analysis of immune responses in urinary bladder epithelial cells, monocyte-like cells and blood mononuclear cells confirm that these waters activate the immune system as well as induce pro- inflammatory responses. The results indicate that the cytokine profiles correlate to the endotoxin load of the waters rather than to the levels of pharmaceuticals or culturable bacteria load, suggesting that measurements of endotoxin levels and immune responses would be a valuable addition to the analysis of inland waters. © 2014 Elsevier B.V. All rights reserved. 1. Introduction Industrialization and population growth has led to increased levels of pollution in the environment, resulting in an elevated risk to human health. The quality of wastewater is of major concern. Chemical and phar- maceutical contaminants have been detected in wastewater treatment plant (WWTP) effluents and inland waters over the years, indicating that the sewage treatment process does not eliminate all substances Science of the Total Environment 488–489 (2014) 228–235 ⁎ Corresponding author. Tel.: +46 19 303943. ⁎⁎ Corresponding author. Tel.: +46 19 301244. E-mail addresses: jana.jass@oru.se (J. Jass), per-erik.olsson@oru.se (P.-E. Olsson). 1 Present address: Biotechnology research center PO Box 3310, Tripoli, Libya. 2 These authors have contributed equally. http://dx.doi.org/10.1016/j.scitotenv.2014.04.090 0048-9697/© 2014 Elsevier B.V. All rights reserved. Contents lists available at ScienceDirect Science of the Total Environment journal homepage: www.elsevier.com/locate/scitotenv