Raptors are ideal indicators for biomonitoring studies using wildlife in order to assess the environmental pollution in the terrestrial ecosystem, since they are placed in the highest trophic position in the food webs and their life expectancy is relatively long. In this study, 26 eggs of 4 bird species (Peregrine falcon, Eurasian curlew, Little owl and Eagle owl) collected in Germany, were investigated for the presence of persistent organic pollutants (POPs) and thousands of contaminants of emerging concern (CECs). Generic sample preparation protocols were followed for the extraction of the analytes and the purification of the extracts, and the samples were analyzed both by liquid (LC) and gas chromatography (GC) coupled to high resolution mass spectrometry (HRMS), for capturing a wide range of organic micropollutants with different physicochemical properties. State-of-the-art screening methodologies were applied in the acquired HRMS data, including wide-scope target analysis of 2448 known pollutants and suspect screening of over 65,000 environmentally relevant compounds. Overall, 58 pollutants from different chemical classes, such as plant protection products, per- and polyfluoroalkyl substances and medicinal products, as well as their transformation products, were determined through target analysis. Most of the detected compounds were lipophilic (logP>2), although the presence of (semi)polar contaminants should not be overlooked, underlying the need for holistic analytical approaches in environmental monitoring studies. p,p'-DDE, PCB 153 and PCB138, PFOS and methylparaben were the most frequently detected compounds. 50 additional substances were identified and semi-quantified through suspect screening workflows, including mainly compounds of industrial use with high production volume.

Discharge of substances like pesticides, pharmaceuticals, flame retardants, and chelating agents in surface waters has increased over the last decades due to the rising numbers of chemicals used by humans and because many WWTPs do not eliminate these substances entirely. The study, results of which are presented here, focused on associations of (1) concentrations of micropollutants in wastewater treatment plant (WWTP) effluents, surface waters, sediments, and tissues of fishes; (2) results of laboratory biotests indicating potentials for effects in these samples and (3) effects either in feral chub (Leuciscus cephalus) from two German rivers (Schussen, Argen) or in brown trout (Salmo trutta f. fario) and rainbow trout (Oncorhynchus mykiss) exposed in bypass systems to streamwater of these rivers or in cages directly in the rivers. The Schussen and Argen Rivers flow into Lake Constance. The Schussen River is polluted by a great number of chemicals, while the Argen River is less influenced by micropollutants. Pesticides, chelating agents, flame retardants, pharmaceuticals, heavy metals, polychlorinated biphenyls (PCBs), and polybrominated diphenyl ethers (PBDEs) were detected in effluents of a WWTP discharging to the Schussen as well as in surface water, and/or fishes from downstream of the WWTP. Results obtained by biotests conducted in the laboratory (genotoxicity, dioxin-like toxicity, and embryotoxicity) were linked to effects in feral fish collected in the vicinity of the WWTP or in fishes exposed in cages or at the bypass systems downstream of the WWTP. Dioxin-like effect potentials detected by reporter gene assays were associated with activation of CYP1A1 enzymes in fishes which are inducible by dioxin-like chemicals. Abundances of several PCBs in tissues of fishes from cages and bypass systems were not associated with these effects but other factors can influence EROD activity. Genotoxic potentials obtained by in vitro tests were associated with the presence of micronuclei in erythrocytes of chub from the river. Chemicals potentially responsible for effects on DNA were identified. Embryotoxic effects on zebrafish (Danio rerio), investigated in the laboratory, were associated with embryotoxic effects in trout exposed in streamwater bypass systems at the two rivers. In general, responses at all levels of organization were more pronounced in samples from the Schussen than in those from the Argen. These results are consistent with the magnitudes of chemical pollution in these two streams. Plausibility chains to establish causality between exposures and effects and to predict effects in biota in the river from studies in the laboratory are discussed.

The project focuses on the efficiency of combined technologies to reduce the release of micropollutants and bacteria into surface waters via sewage treatment plants of different size and via stormwater overflow basins of different types. As a model river in a highly populated catchment area, the river Schussen and, as a control, the river Argen, two tributaries of Lake Constance, Southern Germany, are under investigation in this project. The efficiency of the different cleaning technologies is monitored by a wide range of exposure and effect analyses including chemical and microbiological techniques as well as effect studies ranging from molecules to communities.

The metal platinum is inter alia used for industrial and medical purposes. Due to its application in automobile catalytic converters and as an anti-cancer drug, Pt enters the aquatic environment via road runoff and hospital sewage and raises concerns about its environmental impact and toxicity to organisms. Therefore, the genotoxicity of Pt at 0, 0.1, 1, 10, 50, 100 and 200 μg/l PtCl2 was tested on two freshwater organisms, zebrafish (Danio rerio) and ramshorn snail (Marisa cornuarietis) using the single cell gel electrophoresis, also called comet assay. PtCl2 did not show any genotoxicity for D. rerio at the tested concentrations, whereas significantly elevated DNA damage was observed in M. cornuarietis at 1 μg/l PtCl2 and beyond. The results of the study suggest a high sensitivity of M. cornuarietis concerning the genotoxic impact of PtCl2.

The platinum group metals (PGMs) platinum (Pt), palladium (Pd), and rhodium (Rh) are used in automobile catalytic converters, from which they have been emitted into the environment to an increasing degree during the last 20 years. Despite the bioavailability of these metals to plants and animals, studies determining the effects of PGMs on organisms are extremely rare. In the present study, effects of various concentrations of PtCl2 (0.1, 1, 10, 50 and 100μg/L) were investigated with respect to the induction of hsp70 and histopathological alterations in the zebrafish, Danio rerio and the ramshorn snail, Marisa cornuarietis. Histopathological investigations revealed effects of Pt on both species, which varied between slight and strong cellular reactions, depending on the PtCl2 concentration. The hsp70 level in M. cornuarietis did not show an increase following Pt exposure whereas it was significantly elevated at 100μg/L PtCl2 in D. rerio.

SUMMARY The archetypal body plan of conchiferan molluscs is characterized by an external calcareous shell, though internalization of shells has evolved independently in a number of molluscan clades, including gastropod families. In gastropods, the developmental process of torsion is regarded as a hallmark that is associated with a new anatomical configuration. This configuration is present in extant prosobranch gastropod species, which predominantly bear external shells. Here, we show that short‐term exposure to platinum during development uncouples at least two of the processes associated with torsion of the freshwater snail Marisa cornuarietis. That is, the anus of the treated snails is located anteriorly, but the gill and the designated mantle tissue remains in a posterior location, thus preventing the formation of an external shell. In contrast to the prosobranchian archetype, platinum treatment results in the formation of a posterior gill and a cone‐shaped internal shell, which persists across the lifetime. This first finding of artificially induced snail‐slug conversion was also seen in the pulmonate snail Planorbarius corneus and demonstrates that selective alteration of embryonic key processes can result in fundamental changes of an existing body plan and—if altered regulation is inherited—may give rise to a new one.

Platinum group elements (PGEs), platinum, palladium and rhodium are widely used in automobile catalytic converters. PGEs are emitted into the environment and enter the aquatic ecosystem via runoff rainwater. The present study investigated the bioavailability of platinum chloride for the zebrafish (Danio rerio) and the ramshorn snail (Marisa cornuarietis) and determined the bioaccumulation rate of platinum. Applying the fish early life stage assay for D. rerio (DarT) and the Marisa embryo toxicity test (“Mariett”) for M. cornuarietis, effects of platinum chloride on the embryonic development were investigated. Platinum concentrations tested in this study ranged from environmentally relevant concentrations of 38ngL−1 up to a concentration of 74.2μgL−1 for D. rerio and of 200ngL−1 up to 98.7μgL−1 for M. cornuarietis. Platinum was found to be accumulated in both organisms. Bioaccumulation factors (BAFs) were in the range of 5–55 for D. rerio and of 218.4–723.9 for M. cornuarietis, depending on the tested Pt concentrations. During the embryonic development, platinum was shown to alter the heart rate of both organisms already at the lowest tested concentration. At higher concentrations, platinum decelerated the hatching rate of the embryos of both species. Additionally, a retardation of the general development and a loss of weight due to platinum exposure was observed in M. cornuarietis. Results of this study contribute important data on the ecotoxicity of a rarely studied element.

In this study the effects of 3,4-dichloroaniline (3,4-DCA), a decomposition product of the herbicides propanil and diuron (and other pesticides), and diazinon, a neurotoxic insecticide, on early life stages of zebrafish Danio rerio were assessed. The toxicity of these substances with different modes of action (acetylcholine esterase inhibitor vs. polar narcosis) was tested for single substances as well as in binary mixtures. To study effects on different biological organisation levels (from the molecular up to the whole organism level) the molecular stress response regarding Hsp70, the embryonic and larval development and the locomotor activity were investigated as integrative biomarkers. In single substance tests 3,4-dichloroaniline elicited deformations at ≥0.25 mg/l during the 11 days subchronic test, whereas locomotor activity and mortality were impaired at ≥0.5 mg/l. Diazinon effects on those parameters were obvious at ≥2 mg/l, except for the deformation rate (11 days:1 mg/l). In equitoxic mixtures of both substances concentration additivity was observed for deformation rate and mortality (11 days). An increase in the Hsp70 content occurred in zebrafish exposed to 0.25 mg 3,4-DCA/l as well as to 0.05 mg diazinon/l; in mixtures concentration additivity could be shown. The investigated endpoints varied in respect to their sensitivity, with deformations and Hsp70 levels as most sensitive parameters concerning 3,4-DCA and Hsp70 as most sensitive parameter concerning diazinon. Accordingly, for an integrated understanding of the effects of chemicals and their mixtures on fish, a battery of different test methods should be applied.

The present study examined combined effects of the insecticides thiacloprid and diazinon and a physical stressor, elevated temperature (28, 30 and 33.5 degrees C, control at 26 degrees C), on embryonic development of zebrafish (Danio rerio). Early life stages were exposed to five different concentrations (thiacloprid: 1, 5, 10, 15 and 20mg/L; diazinon: 100, 500, 1000, 2000 and 3000 microg/L) for 72 or 96 h, respectively, depending on the temperature. Thiacloprid was shown to have no effects on early life stage parameters, except on the heart rate. This effect was probably due to an increased metabolism upon exposure to the pollutant. Exposure to diazinon resulted in mortality which strongly increased with elevated temperature. Within the tested temperature range, increased mortality occurred particularly at concentrations of 2000 and 3000 microg/L diazinon. Observed sublethal effects were a decrease of the heart rate with increasing substance concentration, yolk sac edema, heart sac edema and spine deformations, particularly at concentrations of 2000 and 3000 microg/L diazinon. The hatching date was shown to be preponed with increasing temperature, most strikingly in combination with 2000 or 3000 microg/L diazinon. Mixtures of temperature and substance concentration as independent variables were shown to act synergistically in a dose-level-dependent manner on hatching rate, whereas the magnitude of synergism depended on effect levels.

The introduction of organic micropollutants in terrestrial and aquatic ecosystems, which mainly originate from anthropogenic activities, is supported by overwhelming evidence from global studies conducted as early as the turn of the 20th century. A fraction of these xenobiotics, the regulated “Persistent Organic Pollutants (POPs)”, are constantly monitored due to their thoroughly studied persistent, bioaccumulative, and toxic (PBT) properties. Although the occurrence of POPs in the various environmental compartments is well-studied, there are chemicals, which are not systematically subjected to examination, the so-called “Emerging Contaminants (ECs)”. These contaminants are not subjected to marketing restrictions or regulatory investigations but are candidates for future regulation, due to their frequent detection in environmental matrices and their potential toxic properties (Gkotsis et al; 2023, Thomaidis et al; 2012, Badry et al; 2022). The partitioning of organic micropollutants through the ecosystems and their subsequent bioaccumulation and biomagnification in the upper trophic levels necessitate the systematic investigation of their presence in this group of organisms. (Badea et al., 2020; Riget et al., 2016). Subsequently, biomonitoring and risk assessment studies that target apex predators benefit from certain characteristics of their trophic position in the ecosystem, such as their relatively long lifespan over which bioaccumulation occurs, the incorporation of spatial and temporal exposure, and the relative ease of quantifying their populations (Movalli et al; 2022, Malarvannan et al; 2020). Even though there are many biomonitoring data concerning the presence of xenobiotics in marine organisms, the respective data on terrestrial predators are scarce. The aim of the current biomonitoring study is to reveal the chemical fingerprint of terrestrial mesocarnivores gathered from Germany by utilizing complementary chromatographic techniques coupled with High-Resolution Mass Spectrometry (HRMS). In this framework, 23 mesocarnivore liver samples collected in Baden-Württemberg state, Germany between 1993 and 2021, were analyzed following wide-scope target screening methodologies, in order to investigate the presence of organic micropollutants. Different species were analyzed, including European polecats, beech martens, and European pine martens. Briefly, the analytes were extracted from lyophilized liver samples through generic sample preparation protocols using Accelerated Solvent Extraction (ASE), followed by a purification step using Solid Phase Extraction (SPE). The cartridges used for the SPE step were in-house mixed-mode and Florisil, for LC and GC analysis, respectively. The final extracts were analyzed by Liquid Chromatography (LC) with Electrospray Ionization (ESI) and Gas Chromatography (GC) with Atmospheric Pressure Chemical Ionization (APCI) as ion sources, coupled with HRMS, to broaden the chemical domain accessible to wide-scope target analysis. The acquired HR-MS data were screened for the presence of more than 2,400 organic micropollutants, included in the target lists of the National and Kapodistrian University of Athens (NKUA). Compounds of different classes (such as pharmaceuticals, personal care products, illicit drugs, plant protection products, stimulants, industrial chemicals, sweeteners, plasticizers, surfactants, flame retardants, and phthalates), as well as their Transformation products (TPs) and metabolites, are included in the NKUA datasets, which are updated regularly. The LC and GC target lists are available as data sets in Zenodo( (DOI: 10.5281/zenodo.3723478, and GC database DOI:10.5281/zenodo.3753372, respectively). The data treatment protocols applied during the target screening utilized the strict identification thresholds of mass accuracy (<2mDa), retention time shift (<0.20min), isotopic pattern fitting (mSigma<100), and fragmentation pattern match. The standard addition method was used for the quantification of the analytes (Gago-Ferrero et al;2020). The results yielded by the LC & GC HRMS analysis reveal the presence of numerous organic pollutants in the tested samples, classified under several different chemical categories, such as pharmaceuticals (including venlafaxine, gabapentin, oxfendazole), plant protection products (like jasmolin, propoxur, and tralkoxydim), personal care products (for instance the parabens butylparaben, ethylparaben, methylparaben). Furthermore, several industrial chemicals were detected comprising of 4 Polychlorinated Biphenyls (PCBs), 6 Polycyclic Aromatic Hydrocarbons (PAHs) and 14 Per- and Polyfluoroalkyl Substances (PFAS). In addition to the parent compounds, multiple TPs of pharmaceuticals, plant protection products, as well as coffee and tobacco-related compounds were detected in the terrestrial mesocarnivore liver samples. Moreover, the most abundant and frequently detected xenobiotic compound was PFOS, affirming its persistent and bioaccumulative properties already reported in the literature (Rupp et al; 2023). The persistence of these analytes may cause potentially harmful effects on the environment and eventually on human health and therefore should be thoroughly investigated in environmental studies. Overall, the previously indicated findings highlight the importance of HRMS wide-scope target screening methodologies in biomonitoring studies.