Is it worth the extra mile? Comparing environmental DNA and RNA metabarcoding for vertebrate and invertebrate biodiversity surveys in a lowland stream.

Environmental DNA (eDNA) metabarcoding has emerged as a promising approach to assess biodiversity and derive ecological status classes from water samples. However, a limitation of eDNA surveys is that detected DNA molecules may originate from other places or even dead organisms, distorting local biodiversity assessments. Environmental RNA (eRNA) metabarcoding has recently been proposed as a complementary tool for more localized assessments of the biological community. In this study, we evaluated the effectiveness of eDNA and eRNA metabarcoding for inferring the richness and species distribution patterns of vertebrates and invertebrates in a Central European lowland river. We collected water samples and analyzed them using a 12S marker for vertebrates and a COI marker for invertebrates. We detected 31 fish, 16 mammal, 10 bird and one lamprey species in the vertebrate dataset. While results were largely consistent, we detected a higher number of species when analysing eRNA (mean = 30.89) than eDNA (mean = 26.16). Also, eRNA detections had a stronger local signature than eDNA detections when compared against species distribution patterns from traditional fish monitoring data. For invertebrates, we detected 109 arthropod, 22 annelid, 12 rotiferan, eight molluscan and four cnidarian species. In contrast to the pattern of vertebrate richness, we detected a higher richness using eDNA (mean = 41.37) compared to eRNA (mean = 22.42). Our findings primarily show that eDNA and eRNA-based detections are comparable for vertebrate and invertebrate taxa. Biological replication was important for both template molecules studied. Signal detections for vertebrates were more localized for eRNA compared to eDNA. Overall, the advantages of the extra steps needed for eRNA analyses depend on the study question but both methods provide important data for biodiversity monitoring and research.

Network analysis to reveal the most commonly detected compounds in predator-prey pairs in freshwater and marine mammals and fish in Europe.

Marine and freshwater mammalian predators and fish samples, retrieved from environmental specimen banks (ESBs), natural history museum (NHMs) and other scientific collections, were analysed by LIFE APEX partners for a wide range of legacy and emerging contaminants (2545 in total). Network analysis was used to visualize the chemical occurrence data and reveal the predominant chemical mixtures for the freshwater and marine environments. For this purpose, a web tool was created to explore these chemical mixtures in predator-prey pairs. Predominant chemicals, defined as the most prevalent substances detected in prey-predator pairs were identified through this innovative approach. The analysis established the most frequently co-occurring substances in chemical mixtures from AP&P in the marine and freshwater environments. Freshwater and marine environments shared 23 chemicals among their top 25 predominant chemicals. Legacy chemical, including perfluorooctanesulfonic acid (PFOS), brominated diphenyl ethers (BDEs), polychlorinated biphenyls (PCBs), hexachlorobenzene and mercury were dominant chemicals in both environments. Furthermore, N-acetylaminoantipyrine was a predominant pharmaceutical in both environments. The LIFE APEX chemical mixture application (https://norman-data.eu/LIFE_APEX_Mixtures) was proven to be useful to establish most prevalent compounds in terms of number of detected counts in prey-predator pairs. Nonetheless, further research is needed to establish food chain associations of the predominant chemicals.

Metabolites are overlooked in environmental risk assessments and monitoring of pharmaceuticals: The case study of pantoprazole.

The proton-pump inhibitor pantoprazole (PPZ) is one of the most consumed pharmaceuticals worldwide. Despite its high usage, reported PPZ concentrations in environmental water samples are comparatively low, which can be explained by the extensive metabolism of PPZ in the human body. Since most previous studies did not consider human PPZ metabolites it can be assumed that the current environmental exposure associated with the application of PPZ is substantially underestimated. In our study, 4'-O-demethyl-PPZ sulfide (M1) was identified as the predominant PPZ metabolite by analyzing urine of a PPZ consumer as well as the influent and effluent of a wastewater treatment plant (WWTP) using liquid chromatography coupled to high resolution mass spectrometry (LC-HRMS). M1 was found to be ubiquitously present in WWTP effluents (max. concentration: 3 000 ng/L) and surface waters in Germany. On average, the surface water concentrations of M1 were approximately 30 times higher than those of the parent compound PPZ. Laboratory scale experiments demonstrated that activated carbon can considerably adsorb M1 und thus improve its removal during wastewater and drinking water treatment. Laboratory ozonation experiments showed a fast oxidation of M1, accompanied by the formation of several ozonation products. Certain ozonation products (identities confirmed via synthesized reference standards) were also detected in water samples collected after ozonation in a full-scale WWTP. Overall lower signal intensities were observed in the effluents of a sand filter and biologically active granular activated carbon filter, suggesting that the compounds were significantly removed during these post-ozonation treatment stages.

Significant Turning Point: Common Buzzard (Buteo buteo) Exposure to Second-Generation Anticoagulant Rodenticides in the United Kingdom.

Second-generation anticoagulant rodenticides (SGARs) are widely used to control rodent populations, resulting in the serious secondary exposure of predators to these contaminants. In the United Kingdom (UK), professional use and purchase of SGARs were revised in the 2010s. Certain highly toxic SGARs have been authorized since then to be used outdoors around buildings as resistance-breaking chemicals under risk mitigation procedures. However, it is still uncertain whether and how these regulatory changes have influenced the secondary exposure of birds of prey to SGARs. Based on biomonitoring of the UK Common Buzzard (Buteo buteo) collected from 2001 to 2019, we assessed the temporal trend of exposure to SGARs and statistically determined potential turning points. The magnitude of difenacoum decreased over time with a seasonal fluctuation, while the magnitude and prevalence of more toxic brodifacoum, authorized to be used outdoors around buildings after the regulatory changes, increased. The summer of 2016 was statistically identified as a turning point for exposure to brodifacoum and summed SGARs that increased after this point. This time point coincided with the aforementioned regulatory changes. Our findings suggest a possible shift in SGAR use to brodifacoum from difenacoum over the decades, which may pose higher risks of impacts on wildlife.

Temporal trends of lipophilic organic contaminants in blue mussel (1994-2017) and eelpout (1994-2017) from the southern Baltic Sea.

A time-trend study was carried out for two important Baltic Sea species, blue mussel (1994-2017, 11 samples) and eelpout (1994-2017, 11 samples), to track the changes in levels of regulated persistent organic pollutants (POPs) and show potential increases in the levels of the contaminants of emerging concern (CECs). It was carried out utilizing gas chromatography-high-resolution mass spectrometry (GC-HRMS) based non-target screening (NTS). Data were acquired in two modes - electron ionization (EI) and electron capture negative ion chemical ionization (ECNI) - to widen the contaminant coverage, and treated using a fast semi-automated NTS data processing workflow. The study revealed that >250 tentatively identified compounds show statistically significant temporal trends in Baltic blue mussel and eelpout. A large number of regulated substances, including but not limited to PCBs, DDTs and other organochlorine pesticides (OCPs), chlorobenzenes, and many polybrominated diphenyl ethers (PBDEs), showed significant declining trends, as was expected. Their rates of decline were in good agreement with previously reported data. In contrast, increasing trends were observed for many CECs, some polycyclic aromatic compounds (PAHs), and hydrocarbons. The CEC group included, among others, four compounds, namely, one personal care product ingredient, 2-ethylhexyl stearate, one brominated compound 1,2,3,5-tetrabromobenzene and two intermediates 4-isopropoxyaniline and bilobol dimethyl ether, that were reported in marine biota for the first time to the best of our knowledge. Several compounds, including four CECs and two unknown brominated compounds, showed levels considerably higher than the common legacy pollutants (CB-153 and BDE-99), which might be taken into consideration for future monitoring and risk assessment. In addition, this work revealed the presence of a plethora of organoiodinated compounds that exhibited statistically significant temporal trends in the samples under study, which could be of future interest.

Tracking down unknown PFAS pollution - The direct TOP assay in spatial monitoring of surface waters in Germany.

PFAS contamination of surface waters in central Europe was investigated in a spatial monitoring using suspended particulate matter and sediment samples. The samples were collected in 2021 at 171 sampling sites in Germany and at five sites in Dutch waters. All samples were analyzed for 41 different PFAS by target analysis to establish a baseline for these compounds. In addition, a sum parameter approach (direct Total Oxidizable Precursor (dTOP) assay) was used to investigate the PFAS load in the samples more comprehensively. PFAS pollution varied widely between water bodies. Target analysis detected ∑PFAS concentrations between <0.5 and 53.1 µg kg-1 dry weight (dw), while levels of <1.0-337 µg kg-1 dw were determined by dTOP assay. Associations were observed for ∑PFSAdTOP and the percentage of urban area in the vicinity of the sampling sites and, less strongly, for distances to industrial sites (i.e. galvanic/paper) and airports. PFAS hotspots were identified by setting the 90th percentile of the data sets for ∑PFAStarget or ∑PFASdTOP as a threshold. Of the 17 hotspots identified by target analysis or dTOP assay, respectively, there were only six overlaps. Thus, 11 highly contaminated sites could not be identified by classical target analysis. The results demonstrate that target analysis captures only a fraction of the actual PFAS load, while unknown precursors remain undetected. Consequently, if only the results of target analysis are considered in assessments, there is a risk that sites heavily polluted with precursors will not be identified delaying mitigation actions and risking prolonged negative impacts on human health and ecosystems. Moreover, effective PFAS management requires establishing a baseline for PFAS using target and sum parameters such as the dTOP assay, and then monitoring this baseline on a regular basis to support emission control and to inform risk management of its effectiveness.

Long-term trend data for PFAS in soils from German ecosystems, including TOP assay.

In this study, soil top layers from the German Environmental Specimen Bank (ESB) covering a period from 2002 to 2018 and 11 sampling sites representing four ecosystem types were analysed for 24 PFAS (11 PFCA, 7 PFSA, and 6 precursors), including the total oxidizable precursor (TOP) assay. Median ΣPFAS levels before and after TOP assay ranged from 0.31 to 19.7 µg/kg dry weight (dw) and 0.32 to 20.4 µg/kg dw, respectively. The most frequently measured PFAS above LOQ were PFOS and PFOA, which were present at concentrations above 0.1 µg/kg dw in each sample. Both accounted, on average, for half to two-thirds of the total identified PFAS load at most sites. The investigated samples from the near-natural and forestry ecosystem types each had significantly higher PFAS levels than those from the urban-industrial ecosystem type, while samples from the agricultural ecosystem type did not differ distinctly from the others. Increases of median ΣPFAS levels after TOP assay ranged up to slightly >20 %, indicating that precursor compounds do not play a major role in the substance spectrum of German ESB soil samples from different ecosystem types. Total organic carbon (TOC) content in analysed samples ranged from about 1 % in sandy soil to >40 % in litter layer and showed a strong positive correlation with PFAS concentrations. It is therefore highly recommended that TOC content is reported along with PFAS levels in soils. PFAS trend analysis for nine sampling sites before and after TOP assay showed that concentration levels at most sites have remained more or less stable for the sampled period of almost two decades. The constant PFAS levels in soil samples indicate that PFOS and PFOA regulations have not yet had a positive effect on the exposure situation in this environmental compartment.

Long-Term Trends for Blue Mussels from the German Environmental Specimen Bank Show First Evidence of Munition Contaminants Uptake.

Submerged munitions are present in marine waters across the globe. They contain energetic compounds (ECs), such as TNT and metabolites thereof, which are considered carcinogenic, exhibit toxic effects in marine organisms, and may affect human health. The aim of this study was to investigate the occurrence of ECs and their trends in blue mussels from the annual collections of the German Environmental Specimen Bank sampled over the last 30 years at three different locations along the coastline of the Baltic and North Sea. Samples were analyzed by GC-MS/MS for 1,3-dinitrobenzene (1,3-DNB), 2,4-dinitrotoluene (2,4-DNT), 2,4,6-trinitrotoluene (TNT), 2-amino-4,6-dinitrotoluene (2-ADNT), and 4-amino-2,6-dinitrotoluene (4-ADNT). The first signals indicating trace levels of 1,3-DNB were observed in samples from 1999 and 2000. ECs were also found below the limit of detection (LoD) in subsequent years. From 2012 onwards, signals just above the LoD were detected. The highest signal intensities of 2-ADNT and 4-ADNT, just below the LoQ (0.14 ng/g d.w. and 0.17 ng/g d.w., respectively), were measured in 2019 and 2020. This study clearly shows that corroding submerged munitions are gradually releasing ECs into the waters that can be detected in randomly sampled blue mussels, even though the concentrations measured are still in the non-quantifiable trace range.

Quantification and characterization of PFASs in suspended particulate matter (SPM) of German rivers using EOF, dTOPA, (non-)target HRMS.

In this study, we compare analytical methods for PFAS determination-target analysis, non-target screening (NTS), direct total oxidizable precursor assay (dTOPA) and extractable organically bound fluorine (EOF). Therefore, suspended particulate matter (SPM) samples from German rivers at different locations in time series from 2005 to 2020 were analyzed to investigate temporal and spatially resolved trends. In this study 3 PFAS mass balances approaches were utilized: (i) PFAA target vs. PFAS dTOPA, (ii) PFAS target vs. EOF and (iii) PFAS target vs. PFAS dTOPA vs. organofluorines NTS vs. EOF. Mass balance approach (i) revealed high proportions of precursor substances in SPM samples. For the time resolved analysis an increase from 94% (2005) to 97% in 2019 was observable. Also for the spatial resolved analysis precursor proportions were high with >84% at all sampling sites. Mass balance approach (ii) showed that the unidentified EOF (uEOF) fraction increased over time from 82% (2005) to 99% (2019). Furthermore, along the river courses the uEOF increased. In the combined mass balance approach (iii) using 4 different analytical approaches EOF fractions were further unraveled. The EOF pattern was fully explainable at the sampling sites at Saar and Elbe rivers. For the time resolved analysis, an increased proportion of the EOF was now explainable. However, still 27% of the EOF for the time resolved analysis and 25% of the EOF for the spatial resolved analysis remained unknown. Therefore, in a complementary approach, both the EOF and dTOPA reveal unknown gaps in the PFAS mass balance and are valuable contributions to PFAS risk assessment. Further research is needed to identify organofluorines summarized in the EOF parameter.

Berlin statement on legacy and emerging contaminants in polar regions.

Polar regions should be given greater consideration with respect to the monitoring, risk assessment, and management of potentially harmful chemicals, consistent with requirements of the precautionary principle. Protecting the vulnerable polar environments requires (i) raising political and public awareness and (ii) restricting and preventing global emissions of harmful chemicals at their sources. The Berlin Statement is the outcome of an international workshop with representatives of the European Commission, the Arctic Council, the Antarctic Treaty Consultative Meeting, the Stockholm Convention on Persistent Organic Pollutants (POPs), environmental specimen banks, and data centers, as well as scientists from various international research institutions. The statement addresses urgent chemical pollution issues in the polar regions and provides recommendations for improving screening, monitoring, risk assessment, research cooperation, and open data sharing to provide environmental policy makers and chemicals management decision-makers with relevant and reliable contaminant data to better protect the polar environments. The consensus reached at the workshop can be summarized in just two words: "Act now!" Specifically, "Act now!" to reduce the presence and impact of anthropogenic chemical pollution in polar regions by. •Establishing participatory co-development frameworks in a permanent multi-disciplinary platform for Arctic-Antarctic collaborations and establishing exchanges between the Arctic Monitoring and Assessment Program (AMAP) of the Arctic Council and the Antarctic Monitoring and Assessment Program (AnMAP) of the Scientific Committee on Antarctic Research (SCAR) to increase the visibility and exchange of contaminant data and to support the development of harmonized monitoring programs. •Integrating environmental specimen banking, innovative screening approaches and archiving systems, to provide opportunities for improved assessment of contaminants to protect polar regions.

Differences in the internal PFAS patterns of herbivores, omnivores and carnivores - lessons learned from target screening and the total oxidizable precursor assay.

Per- and polyfluorinated alkyl substances (PFAS) are a group of anthropogenic chemicals, which are not (fully) biodegradable and accumulate in different environmental compartments worldwide. A comprehensive, quantitative analysis - consisting of target analysis (66 different analytes, including e. g. ultrashort-chain perfluorinated carboxylic acids (PFCAs), precursor compounds and novel substitutes) and the Total Oxidisable Precursor (TOP) assay (including trifluoroacetic acid (TFA)) - were conducted to analyse the PFAS concentrations and patterns in 12 mammalian and two bird species from different areas of Germany and Denmark. The PFAS contamination was investigated in dependance of the trophic class (herbivores, omnivores, carnivores), ecological habitat (terrestrial, (semi-) aquatic) and body tissue (liver, musculature). PFAS concentrations were highest in carnivores, followed by omnivores and herbivores, with ∑PFAS concentration ranging from 1274 µg/kg (Eurasian otter liver) to 22 µg/kg (roe deer liver). TFA dominated in the herbivorous species, whereas perfluorooctanesulfonic acid (PFOS) and the long-chain PFCAs covered the majority of the PFAS contamination in carnivorous species. Besides trophic class, ecological habitat also affected the PFAS levels in the different species, with terrestrial herbivores and omnivores showing higher PFAS concentration than their aquatic counterparts, whereas for carnivores this relationship was reversed. The TOP assay analysis indicated similar trends, with the PFCA formation pattern differing significantly between the trophic classes. TFA was formed predominantly in herbivorous and omnivorous species, whereas in carnivorous species a broad spectrum of PFCAs (chain-length C2-C14) was formed. Musculature tissue of six species exhibited significantly lower PFAS concentrations than the respective liver tissue, but with similar PFAS patterns. The comprehensive approach applied in the present study showed, that primarily the trophic class is decisive for the PFAS concentration, as herbivores, omnivores and carnivores clearly differed in their PFAS concentrations and patterns. Additionally, the TOP assay gave novel insights in the PFCA formation potential in biota samples.

The importance of in-year seasonal fluctuations for biomonitoring of apex predators: A case study of 14 essential and non-essential elements in the liver of the common buzzard (Buteobuteo) in the United Kingdom.

Trace elements are chemical contaminants spread in the environment by anthropogenic activities and threaten wildlife and human health. Many studies have investigated this contamination in apex raptors as sentinel birds. However, there is limited data for long-term biomonitoring of multiple trace elements in raptors. In the present study, we measured the concentrations of 14 essential and non-essential trace elements in the livers of the common buzzard (Buteo buteo) collected in the United Kingdom from 2001 to 2019 and investigated whether concentrations have changed during this period. In addition, we estimated the importance of selected variables for modelling element accumulations in tissues. Except for cadmium, hepatic concentrations of harmful elements in most buzzards were lower than the biological significance level of each element. Hepatic concentrations of certain elements, including lead, cadmium, and arsenic, varied markedly seasonally within years. Their peak was in late winter and trough in late summer, except copper which showed an opposite seasonal pattern. In addition, lead in the liver consistently increased over time, whereas strontium showed a decreasing trend. Hepatic concentrations of cadmium, mercury, and chromium increased with age, whereas selenium and chromium were influenced by sex. Hepatic concentrations of arsenic and chromium also differed between different regions. Overall, our samples showed a low risk of harmful effects of most elements compared to the thresholds reported in the literature. Seasonal fluctuation was an important descriptor of exposure, which might be related to the diet of the buzzard, the ecology of their prey, and human activities such as the use of lead shot for hunting. However, elucidating reasons for these observed trends needs further examination, and biomonitoring studies exploring the effects of variables such as age, sex, and seasonality are required.

Identification and trend analysis of organic cationic contaminants via non-target screening in suspended particulate matter of the German rivers Rhine and Saar.

Non-target screening of suspended particulate matter (SPM), collected from the German rivers Rhine and Saar, was conducted with the goal of identifying organic, permanent cationic contaminants and of estimating their temporal trends over an extended period. Therefore, annual composite samples of SPM, provided by the German Environmental Specimen Bank, were extracted and analyzed with high resolution LC-QToF-MS/MS. To facilitate the identification of substances belonging to the class "permanent cations", prioritization methods were applied utilizing the physicochemical properties of these compounds. These methods include both interactions of the analyte molecules with cation exchange resins and analyzing mass deviations when changing from non-deuterated to deuterated mobile phase solvents during LC-MS analysis. By applying both methods in a combined approach, 123 of the initially detected 2695 features were prioritized, corresponding to a 95% data reduction. This led to the identification of 22 permanent cationic species. The organic dyes Basic Yellow 28 and Fluorescent Brightener 363 as well as two quaternary ammonium compounds (QACs) were detected in environmental samples for the first time to best of or knowledge. The other compounds include additional QACs, as well as quaternary tri-phenylphosphonium compounds (QPC/TPP). In addition to identification, we determined temporal trends of all compounds over a period of 13 years and assessed their ecotoxicological relevance based on estimated concentrations. The two QACs oleyltrimethylammonium and eicosyltrimethylammonium show significant increasing trends in the Rhine SPM and maximum concentrations in the Saar SPM of about 900 and 1400 µg/kg, respectively. In the case of the dyes, constant trends have been observed at the end of the studied period, but also maximum concentrations of 400 µg/kg for Basic Yellow 28 in 2006 and 1000 µg/kg for Fluorescent Brightener 363 in 2015, potentially indicating a strong ecotoxicological risk.

Assessment of contaminants of emerging concern in European apex predators and their prey by LC-QToF MS wide-scope target analysis.

Apex predators are good indicators of environmental pollution since they are relatively long-lived and their high trophic position and spatiotemporal exposure to chemicals provides insights into the persistent, bioaccumulative and toxic (PBT) properties of chemicals. Although monitoring data from apex predators can considerably support chemicals' management, there is a lack of pan-European studies, and longer-term monitoring of chemicals in organisms from higher trophic levels. The present study investigated the occurrence of contaminants of emerging concern (CECs) in 67 freshwater, marine and terrestrial apex predators and in freshwater and marine prey, gathered from four European countries. Generic sample preparation protocols for the extraction of CECs with a broad range of physicochemical properties and the purification of the extracts were used. The analysis was performed utilizing liquid (LC) chromatography coupled to high resolution mass spectrometry (HRMS), while the acquired chromatograms were screened for the presence of more than 2,200 CECs through wide-scope target analysis. In total, 145 CECs were determined in the apex predator and their prey samples belonging in different categories, such as pharmaceuticals, plant protection products, per- and polyfluoroalkyl substances, their metabolites and transformation products. Higher concentration levels were measured in predators compared to prey, suggesting that biomagnification of chemicals through the food chain occurs. The compounds were prioritized for further regulatory risk assessment based on their frequency of detection and their concentration levels. The majority of the prioritized CECs were lipophilic, although the presence of more polar contaminants should not be neglected. This indicates that holistic analytical approaches are required to fully characterize the chemical universe of biota samples. Therefore, the present survey is an attempt to systematically investigate the presence of thousands of chemicals at a European level, aiming to use these data for better chemicals management and contribute to EU Zero Pollution Ambition.

Environmental DNA from archived leaves reveals widespread temporal turnover and biotic homogenization in forest arthropod communities.

A major limitation of current reports on insect declines is the lack of standardized, long-term, and taxonomically broad time series. Here, we demonstrate the utility of environmental DNA from archived leaf material to characterize plant-associated arthropod communities. We base our work on several multi-decadal leaf time series from tree canopies in four land use types, which were sampled as part of a long-term environmental monitoring program across Germany. Using these highly standardized and well-preserved samples, we analyze temporal changes in communities of several thousand arthropod species belonging to 23 orders using metabarcoding and quantitative PCR. Our data do not support widespread declines of α-diversity or genetic variation within sites. Instead, we find a gradual community turnover, which results in temporal and spatial biotic homogenization, across all land use types and all arthropod orders. Our results suggest that insect decline is more complex than mere α-diversity loss, but can be driven by ß-diversity decay across space and time.

Insects are a barometer of environmental health. Ecosystems around the world are being subjected to unprecedented man-made stresses, ranging from climate change to pollution and intensive land use. These stresses have been associated with several recent, dramatic declines in insect populations, particularly in areas with heavily industrialised farming practices. Despite this, the links between insect decline, environmental stress, and ecosystem health are still poorly-understood. A decline in one area might look catastrophic, but could simply be part of normal, longer-term variations. Often, we do not know whether insect decline is a local phenomenon or reflects wider environmental trends. Additionally, most studies do not go far back enough in time or cover a wide enough geographical range to make these distinctions. To understand and combat insect decline, we therefore need reliable methods to monitor insect populations over long periods of time. To solve this problem, Krehenwinkel, Weber et al. gathered data on insect communities from a new source: tree leaves. Originally, these samples were collected to study air pollution, but they also happen to contain the DNA of insects that interacted with them before they were collected ­ for example, DNA deposited in chew marks where the insects had nibbled on the leaves. This is called environmental DNA, or eDNA for short. To survey the insect communities that lived in these trees, Krehenwinkel, Weber et al. first extracted eDNA from the leaves and sequenced it. Analysis of the different DNA sequences from the leaf samples revealed not only the number of insect species, but also the abundance (or rarity) of each species within each community. Importantly, the leaves had been collected and stored in stable conditions over several decades, allowing changes in these insect populations to be tracked over time. eDNA analysis revealed subtle changes in the make-up of forest insect communities. In the forests where the leaves were collected, the total number of insect species remained much the same over time. However, many individual species still declined, only to be replaced by newcomer species. These 'colonisers' are also widespread, which will likely lead to an overall pattern of fewer species that are more widely distributed ­ in other words, more homogeneity. The approach of Krehenwinkel, Weber et al. provides a reliable method to study insect populations in detail, over multiple decades, using archived samples from environmental studies. The information gained from this has real-world significance for environmental issues with enormous social impact, ranging from conservation, to agriculture and even public health.

The NORMAN Suspect List Exchange (NORMAN-SLE): facilitating European and worldwide collaboration on suspect screening in high resolution mass spectrometry.

Background: The NORMAN Association (https://www.norman-network.com/) initiated the NORMAN Suspect List Exchange (NORMAN-SLE; https://www.norman-network.com/nds/SLE/) in 2015, following the NORMAN collaborative trial on non-target screening of environmental water samples by mass spectrometry. Since then, this exchange of information on chemicals that are expected to occur in the environment, along with the accompanying expert knowledge and references, has become a valuable knowledge base for "suspect screening" lists. The NORMAN-SLE now serves as a FAIR (Findable, Accessible, Interoperable, Reusable) chemical information resource worldwide. Results: The NORMAN-SLE contains 99 separate suspect list collections (as of May 2022) from over 70 contributors around the world, totalling over 100,000 unique substances. The substance classes include per- and polyfluoroalkyl substances (PFAS), pharmaceuticals, pesticides, natural toxins, high production volume substances covered under the European REACH regulation (EC: 1272/2008), priority contaminants of emerging concern (CECs) and regulatory lists from NORMAN partners. Several lists focus on transformation products (TPs) and complex features detected in the environment with various levels of provenance and structural information. Each list is available for separate download. The merged, curated collection is also available as the NORMAN Substance Database (NORMAN SusDat). Both the NORMAN-SLE and NORMAN SusDat are integrated within the NORMAN Database System (NDS). The individual NORMAN-SLE lists receive digital object identifiers (DOIs) and traceable versioning via a Zenodo community (https://zenodo.org/communities/norman-sle), with a total of > 40,000 unique views, > 50,000 unique downloads and 40 citations (May 2022). NORMAN-SLE content is progressively integrated into large open chemical databases such as PubChem (https://pubchem.ncbi.nlm.nih.gov/) and the US EPA's CompTox Chemicals Dashboard (https://comptox.epa.gov/dashboard/), enabling further access to these lists, along with the additional functionality and calculated properties these resources offer. PubChem has also integrated significant annotation content from the NORMAN-SLE, including a classification browser (https://pubchem.ncbi.nlm.nih.gov/classification/#hid=101). Conclusions: The NORMAN-SLE offers a specialized service for hosting suspect screening lists of relevance for the environmental community in an open, FAIR manner that allows integration with other major chemical resources. These efforts foster the exchange of information between scientists and regulators, supporting the paradigm shift to the "one substance, one assessment" approach. New submissions are welcome via the contacts provided on the NORMAN-SLE website (https://www.norman-network.com/nds/SLE/). Supplementary Information: The online version contains supplementary material available at 10.1186/s12302-022-00680-6.

Environmental specimen banks and the European Green Deal.

The study highlights the potential of Environmental Specimen Banks (ESBs) for implementing the Zero Pollution Ambition and the Biodiversity Strategy of the European Green Deal. By drawing on recent monitoring studies of European ESBs, we illustrate the role ESBs already play in assessing the state of ecosystems in Europe and how they help to make developments over time visible. The studies reveal the ubiquitous presence of per- and polyfluoroalkyl substances, halogenated flame retardants, chlorinated paraffins, plasticizers, cyclic volatile methyl siloxanes, UV-filters, pharmaceuticals, and microplastics in the European environment. Temporal trends demonstrate the effectiveness of European regulations on perfluorooctane sulfonic acid, pentabrominated diphenylethers and diethylhexyl phthalate, but also point to the rise of substitutes such as non-phthalate plasticizers and short-chain perfluoroalkyl substances. Other studies are wake-up calls indicating the emergence of currently unregulated compounds such as long-chain chlorinated paraffins. Ecological studies show temporal trends in biometric parameters and stable isotope signatures that suggest long-term changes in environmental conditions. Studies on biodiversity of ecosystems using environmental DNA are still in their beginnings, but here too there is evidence of shifts in community composition that can be linked to changing environmental conditions. This review demonstrates the value of ESBs (a) for describing the status of the environment, (b) for monitoring temporal changes in environmental pollution and the ecologic condition of ecosystems and thereby (c) for supporting regulators in prioritizing their actions towards the objectives of the Green Deal.

Making use of apex predator sample collections: an integrated workflow for quality assured sample processing, analysis and digital sample freezing of archived samples.

Using monitoring data from apex predators for chemicals risk assessment can provide important information on bioaccumulating as well as biomagnifying chemicals in food webs. A survey among European institutions involved in chemical risk assessment on their experiences with apex predator data in chemical risk assessment revealed great interest in using such data. However, the respondents indicated that constraints were related to expected high costs, lack of standardisation and harmonised quality criteria for exposure assessment, data access, and regulatory acceptance/application. During the Life APEX project, we demonstrated that European sample collections (i.e. environmental specimen banks (ESBs), research collection (RCs), natural history museums (NHMs)) archive a large variety of biological samples that can be readily used for chemical analysis once appropriate quality assurance/control (QA/QC) measures have been developed and implemented. We therefore issued a second survey on sampling, processing and archiving procedures in European sample collections to derive key quality QA/QC criteria for chemical analysis. The survey revealed great differences in QA/QC measures between ESBs, NHMs and RCs. Whereas basic information such as sampling location, date and biometric data were mostly available across institutions, protocols to accompany the sampling strategy with respect to chemical analysis were only available for ESBs. For RCs, the applied QA/QC measures vary with the respective research question, whereas NHMs are generally less aware of e.g. chemical cross-contamination issues. Based on the survey we derived key indicators for assessing the quality of biota samples that can be easily implemented in online databases. Furthermore, we provide a QA/QC workflow not only for sampling and processing but also for the chemical analysis of biota samples. We focussed on comprehensive analytical techniques such as non-target screening and provided insights into subsequent storage of high-resolution chromatograms in online databases (i.e. digital sample freezing platform) to ultimately support chemicals risk assessment.

Comprehensive screening of polar emerging organic contaminants including PFASs and evaluation of the trophic transfer behavior in a freshwater food web.

Bioaccumulation and trophic transfer of persistent legacy contaminants have been intensively characterized, but little is known on the contaminants of emerging concern (CECs) in freshwater food webs. Herein, we comprehensively screened CECs with a focus on polar substances and further evaluated their trophic transfer behavior in selected items from the food web of Lake Templin, Germany. Weselected one plankton, two mussel, and nine fish samples covering three trophic levels. With an effective multi-residue sample preparation method and high-resolution mass spectrometry-based target, suspect, and non-target screening, we characterized 477 targets and further screened unknown features in complex biota matrices. Of the 477 targets, 145 were detected and quantified in at least one species (0.02-3640 ng/g, dry weight). Additionally, the suspect and non-target analysis with experimental mass spectra libraries and in silico techniques (MetFrag and SIRIUS4/CSI:FingerID) enabled further identification of 27 unknown compounds with 19 confirmed by reference standards. Overall, the detected compounds belong to a diverse group of chemicals, including 71 pharmaceuticals, 27 metabolites, 26 pesticides, 16 per- and polyfluoroalkyl substances (PFASs), 4 plasticizers, 3 flame retardants, 11 other industrial chemicals and 14 others. Moreover, we determined the trophic magnification factor (TMF) of 34 polar CECs with >80% detection frequency, among which 6 PFASs including perfluorooctane sulfonic acid (PFOS), perfluorodecanoic acid (PFDA), perfluorohexane sulfonic acid (PFHxS), perfluorotridecanoic acid (PFTrA), perfluorotetradecanoic acid (PFTeA), and perfluoroundecanoic acid (PFUnA), exhibited biomagnification potential (TMF =1.8 - 4.2, p < 0.05), whereas 5 pharmaceuticals (phenazone, progesterone, venlafaxine, levamisole, and lidocaine) and 1 personal care product metabolite (galaxolidone) showed biodilution potential (TMF = 0.4 - 0.6, p < 0.05).

30 years trends of microplastic pollution: Mass-quantitative analysis of archived mussel samples from the North and Baltic Seas.

Microplastics (MP) are ubiquitous throughout the environment as a result of an ongoing, increasing, but also lavish use, of plastics over time and its inherent persistence. In contrast, there are almost no data that allow drawing conclusions about the evolution of plastic pollution in the environment over the past decades. This study investigates the MP load in blue mussels from the North and Baltic Sea archived by the German Environmental Specimen Bank in a time series covering almost 30 years. Samples were enzymatically and chemically oxidative digested for MP extraction and subsequent analyzed mass-quantitatively for nine common polymer clusters by pyrolysis gas chromatography-mass spectrometry. Seven polymer clusters were detected in mussel tissue. Summed MP levels were at ppm levels (<20 µg/g mussel, dry weight). North Sea samples reflected a gradual increase from the 1980s/90s to the 2000s whereas those from Baltic Sea showed consistently higher, rather constant MP levels similar to the North Sea site later than 2000. Polymer composition of both sites stood out by cluster (C) of C-PVC and C-PET at both sites. Mussels from Baltic Sea site had larger C-PE and C-PP proportions. Opposed polymer- and site-specific trends indicated both regional and trans-regional MP sources for different polymer clusters. The MP composition of mussels showed strong similarities with adjacent sediment and water samples. The study introduces a relevant dataset addressing the temporal development of MP pollution. It emphasizes a high indicative potential of environmental MP composition/loads received by mussels but raises the necessity on adequate control materials accompany such kind of studies.