Pollutant load and ecotoxicological effects of sediment from stormwater retention basins to receiving surface water on Lumbriculus variegatus.

The overflow of stormwater retention basins during intense and prolonged precipitation events may result in the direct input of particulate pollutants and remobilization of already sedimented particle-bound pollutants to receiving freshwater bodies. Particle-bound pollutants may cause adverse effects on aquatic biota, particularly sediment dwellers. Therefore, we investigated the sediment pollution load of a stream connected to the outfalls of two stormwater basins to determine the impact of the basins' discharges on the metal and organic pollutant content of the sediment. Also, the possible adverse effects of the pollutant load on benthic dwellers were evaluated in sediment toxicity tests with Lumbriculus variegatus and the effects on its growth, reproduction and the biomarkers catalase, acetylcholinesterase and metallothionein were analyzed. The results showed that the retention basins contained the highest load of pollutants. The pollutant load in the stream did not show a clear pollution pattern from the inlets. However, metal enrichment ratios revealed contamination with Cu, Pb and Zn with Pb and Zn above threshold effect concentrations in all sites. Ecotoxicity results showed that the retention basin samples were the most toxic compared to sediment from the stream. Exposure experiments with the stream sediment did not show considerable effects on reproduction, catalase activity and metallothionein concentration. However, modest inhibitions of growth and activity of acetylcholinesterase were detected. Based on the observed results, it cannot be concluded that overflows of the retention basin are responsible for the pollutant contents downstream of their inlet. Other sources that were not considered in this study, such as diffuse input, historic pollution and point sources further upstream as well as along the stream, are likely the major contributors of pollutant load in the sediment of the studied transects of the stream. Additionally, the observed results in the stormwater basin sediment further highlight their importance in retaining particle-bound pollutants and preventing ecotoxicological effects from receiving surface water bodies.

High-performance thin-layer chromatography in combination with an acetylcholinesterase-inhibition bioassay with pre-oxidation of organothiophosphates to determine neurotoxic effects in storm, waste, and surface water.

Pesticides such as organothiophosphates (OTPs) are neurotoxically active and enter the aquatic environment. Bioassays, using acetylcholinesterase (AChE), a suitable substrate and reactant, can be applied for the photometric detection of AChE-inhibiton (AChE-I) effects. The oxidized forms of OTPs, so-called oxons, have higher inhibition potentials for AChE. Therefore, a higher sensitivity is achieved for application of oxidized samples to the AChE assay. In this study, the oxidation of malathion, parathion, and chlorpyrifos by n-bromosuccinimide (NBS) was investigated in an approach combining high-performance thin-layer chromatography (HPTLC) with an AChE-I assay. Two AChE application approaches, immersion and spraying, were compared regarding sensitivity, precision, and general feasibility of the OTP effect detection. The oxidation by NBS led to an activation of the OTPs and a strong increase in sensitivity similar to the oxons tested. The sensitivity and precision of the two application techniques were similar, although the spray method was slightly more sensitive to the oxidized OTPs. The 10% inhibition concentrations (IC10) for the spray approach were 0.26, 0.75, and 0.35 ng/spot for activated malathion, parathion, and chlorpyrifos, respectively. AChE-I effect recoveries in samples from a stormwater retention basin and receiving stream were between 69 and 92% for malathion, parathion, and chlorpyrifos. The overall workflow, including sample enrichment by solid-phase extraction, HPTLC, oxidation of OTPs, and AChE-I assay, was demonstrated to be suitable for the detection of AChE-I effects in native water samples. An effect of unknown origin was found in a sample from a stormwater retention basin.

High-performance thin-layer chromatography in combination with a yeast-based multi-effect bioassay to determine endocrine effects in environmental samples.

Effect-directed analysis (EDA) that combines effect-based methods (EBMs) with high-performance thin-layer chromatography (HPTLC) is a useful technique for spatial, temporal, and process-related effect evaluation and may provide a link between effect testing and responsible substance identification. In this study, a yeast multi endocrine-effect screen (YMEES) for the detection of endocrine effects is combined with HPTLC. Simultaneous detection of estrogenic, androgenic, and gestagenic effects on the HPTLC plate is achieved by mixing different genetically modified Arxula adeninivorans yeast strains, which contain either the human estrogen, androgen, or progesterone receptor. Depending on the yeast strain, different fluorescent proteins are formed when an appropriate substance binds to the specific hormone receptor. This allows to measure hormonal effects at different wavelengths. Two yeast cell application approaches, immersion and spraying, are compared. The sensitivity and reproducibility of the method are shown by dose-response investigations for reference compounds. The spraying approach indicated similar sensitivities and higher precisions for the tested hormones compared to immersion. The EC10s for estrone (E1), 17ß-estradiol (E2), 17α-ethinylestradiol (EE2), 5α-dihydrotestosterone (DHT), and progesterone (P4) were 95, 1.4, 10, 7.4, and 15 pg/spot, respectively. Recovery rates of E1, E2, EE2, DHT, and P4 between 88 and 120% show the usability of the general method in combination with sample enrichment by solid phase extraction (SPE). The simultaneous detection of estrogenic, androgenic, and gestagenic effects in wastewater and surface water samples demonstrates the successful application of the YMEES in such matrices. This promising method allows us to identify more than one endocrine effect on the same HPTLC plate, which saves time and material. The method could be used for comparison, evaluation, and monitoring of different river sites and wastewater treatment steps and should be tested in further studies.

Comparison of Software Tools for Liquid Chromatography-High-Resolution Mass Spectrometry Data Processing in Nontarget Screening of Environmental Samples.

The field of high-resolution mass spectrometry has undergone a rapid progress in the last years due to instrumental improvements leading to a higher sensitivity and selectivity of instruments. A variety of qualitative screening approaches, summarized as nontarget screening, have been introduced and have successfully extended the environmental monitoring of organic micropollutants. Several automated data processing workflows have been developed to handle the immense amount of data that are recorded in short time frames by these methods. Most data processing workflows include similar steps, but underlying algorithms and implementation of different processing steps vary. In this study the consistency of data processing with different software tools was investigated. For this purpose, the same raw data files were processed with the software packages MZmine2, enviMass, Compound Discoverer, and XCMS online and resulting feature lists were compared. Results show a low coherence between different processing tools, as overlap of features between all four programs was around 10%, and for each software between 40% and 55% of features did not match with any other program. The implementation of replicate and blank filter was identified as one of the sources of observed divergences. However, there is a need for a better understanding and user instructions on the influence of different algorithms and settings on feature extraction and following filtering steps. In future studies it would be of interest to investigate how final data interpretation is influenced by different processing software. With this work we want to encourage more awareness on data processing as a crucial step in the workflow of nontarget screening.

Evaluation of a biological post-treatment after full-scale ozonation at a municipal wastewater treatment plant.

To reduce the discharge of trace organic compounds into water bodies associated with potential toxic effects such as endocrine disruption, new advanced treatment methods are being investigated at several wastewater treatment plants (WWTPs). One of the most studied and already implemented technologies is ozonation. However, ozonation only partially oxidizes trace organic compounds (TrOC) and as a result, transformation products (TPs) with unknown properties can be formed. In order to minimise the risk of releasing unknown and potentially toxic TPs into surface water, it is recommended to install a biological post-treatment after ozonation. The aim of this study was to evaluate the efficiency of a moving bed reactor following ozonation in a full-scale plant. Different ozone dosages (zspec. = 0.3, 0.5, 0.7 mg O3/mgDOC) were investigated. To assess the biological activity of the post-treatment, the assimilable organic carbon (AOC) was determined in addition to the formed biomass. Furthermore, selected TrOC were analysed in parallel to monitor the ozonation efficiency at different ozone doses. In addition, estrogenic, androgenic as well as corresponding antagonistic effects were investigated after each treatment step using the A-YES and A-YAS assay. A non-target screening was performed to evaluate a trend analysis of formed TPs as well as their removal by the post-treatment procedure. The results proved the successful design of the biological post-treatment reactor by a constant biofilm development and reduction of the AOC. Endocrine effects were removed below the limit of detection (LOD) of 10 pg EEQ/L already after ozonation for all applied ozone doses. Antagonistic effects were not significantly reduced during ozonation and subsequent biological post-treatment. For this reason, further research is needed to evaluate different post-treatment technologies. The trend analysis from non-target screening data showed a reduction of about 95% of the number of formed TPs by the biological post-treatment. Consequently, an assessment of the biological activity and the elimination capacity of a certain biological post-treatment technique is thus possible by applying the AOC in combination with a non-target screening.