Characterization of wastewater effluents in the Danube River Basin with chemical screening, in vitro bioassays and antibiotic resistant genes analysis

Nikiforos A Alygizakis,Harrie Besselink,Gabriela K Paulus,Peter Oswald,Luc M Hornstra,Martina Oswaldova,Gertjan Medema,Nikolaos S Thomaidis,Peter A Behnisch,Jaroslav Slobodnik

doi:10.1016/j.envint.2019.03.060

Abstract

Averaged 7-day composite effluent wastewater samples from twelve wastewater treatment plants (WWTPs) in nine countries (Romania, Serbia, Hungary, Slovenia, Croatia, Slovakia, Czechia, Austria, Germany) in the Danube River Basin were collected. WWTPs' selection was based on countries' dominant technology and a number of served population with the aim to get a representative holistic view of the pollution status. Samples were analyzed for 2248 chemicals of emerging concern (CECs) by wide-scope target screening employing LC-ESI-QTOF-MS. 280 compounds were detected at least in one sample and quantified. Spatial differences in the concentrations and distribution of the compounds classes were discussed. Additionally, samples were analyzed for the possible agonistic/antagonistic potencies using a panel of in vitro transactivation reporter gene CALUX® bioassays including ERα (estrogenics), anti-AR (anti-androgens), GR (glucocorticoids), anti-PR (anti-progestins), PPARα and PPARγ (peroxisome proliferators) and PAH assays. The potency of the wastewater samples to cause oxidative stress and induce xenobiotic metabolism was determined using the Nrf2 and PXR CALUX® bioassays, respectively. The signals from each of the bioassays were compared with the recently developed effect-based trigger values (EBTs) and thus allowed for allocating the wastewater effluents into four categories based on their measured toxicity, proposing a putative action plan for wastewater operators. Moreover, samples were analyzed for antibiotics and 13 antibiotic-resistant genes (ARGs) and one mobile genetic element (intl1) with the aim to assess the potential for antibiotic resistance. All data collected from these various types of analysis were stored in an on-line database and can be viewed via interactive map at https://norman-data.eu/EWW_DANUBE.

Highlights

The Danube River Basin (DRB) is the world's most international river basin covering a total area of 801,463 km2, including territories from 19 countries
2 L from the weekly averaged samples were processed for chemical analysis, 0.5 L were sent to KWR (Nieuwegein, Netherlands) for analysis of antibiotic resistant genes (ARGs) and 0.5 L were sent to BDS (Amsterdam, Netherlands) for application of bioassays
Representative effluent wastewater samples were collected from nine countries of DRB

Summary

Introduction

The Danube River Basin (DRB) is the world's most international river basin covering a total area of 801,463 km, including territories from 19 countries. Careful management of DRB's water resources is needed, including control over chemical pollution. European Union (EU) environmental legislation aims to protect all European water bodies by achieving their good chemical and ecological status (European Commission, 2013). In the content of chemical status, the EU Water Framework Directive (WFD) established a list of 45 priority substances (European Commission, 2013), supplemented by a set of additional 15 compounds (European Commission, 2018) in the recently revised Watch list, which are required to be monitored by Member States and benchmark their concentration against the Environmental Quality Standards (EQS). Many toxic anthropogenic chemicals are released into the environment that may have an adverse effects on the human health, ecosystem and diminish the quality of the aquatic resources (Altenburger et al, 2015).

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