Bioaccessibility of Organic Compounds Associated with Tire Particles Using a Fish In Vitro Digestive Model: Solubilization Kinetics and Effects of Food Coingestion.

Tire and road wear particles (TRWP) account for an important part of the polymer particles released into the environment. There are scientific knowledge gaps as to the potential bioaccessibility of chemicals associated with TRWP to aquatic organisms. This study investigated the solubilization and bioaccessibility of seven of the most widely used tire-associated organic chemicals and four of their degradation products from cryogenically milled tire tread (CMTT) into fish digestive fluids using an in vitro digestion model based on Oncorhynchus mykiss. Our results showed that 0.06-44.1% of the selected compounds were rapidly solubilized into simulated gastric and intestinal fluids within a typical gut transit time for fish (3 h in gastric and 24 h in intestinal fluids). The environmentally realistic scenario of coingestion of CMTT and fish prey was explored using ground Gammarus pulex. Coingestion caused compound-specific changes in solubilization, either increasing or decreasing the compounds' bioaccessibility in simulated gut fluids compared to CMTT alone. Our results emphasize that tire-associated compounds become accessible in a digestive milieu and should be studied further with respect to their bioaccumulation and toxicological effects upon passage of intestinal epithelial cells.

In Vitro Digestion of Tire Particles in a Fish Model (Oncorhynchus mykiss): Solubilization Kinetics of Heavy Metals and Effects of Food Coingestion.

Tire and road wear particles (TRWP) have been shown to represent a large part of anthropogenic particles released into the environment. Nevertheless, the potential ecological risk of TRWP in the different environmental compartments and their potential toxic impacts on terrestrial and aquatic organisms remain largely underinvestigated. Several heavy metals compose TRWP, including Zn, which is used as a catalyst during the vulcanization process of rubber. This study investigated the solubilization potential of metals from cryogenically milled tire tread (CMTT) and TRWP in simulated gastric fluids (SFGASTRIC) and simulated intestinal fluids (SFINTESTINAL) designed to mimic rainbow trout (Oncorhynchus mykiss) gastrointestinal conditions. Our results indicate that the solubilization of heavy metals was greatly enhanced by gastrointestinal fluids compared to that by mineral water. After a 26 h in vitro digestion, 9.6 and 23.0% of total Zn content of CMTT and TRWP, respectively, were solubilized into the simulated gastrointestinal fluids. Coingestion of tire particles (performed with CMTT only) and surrogate prey items (Gammarus pulex) demonstrated that the animal organic matter reduced the amount of bioavailable Zn solubilized from CMTT. Contrastingly, in the coingestion scenario with vegetal organic matter (Lemna minor), high quantities of Zn were solubilized from L. minor and cumulated with Zn solubilized from CMTT.

Role of Biofilms in Contaminant Bioaccumulation and Trophic Transfer in Aquatic Ecosystems: Current State of Knowledge and Future Challenges.

In freshwater environments, microbial assemblages attached to submerged substrates play an essential role in ecosystem processes such as primary production, supported by periphyton, or organic matter decomposition, supported by microbial communities attached to leaf litter or sediments. These microbial assemblages, also called biofilms, are not only involved in nutrients fluxes but also in contaminants dynamics. Biofilms can accumulate metals and organic contaminants transported by the water flow and/or adsorbed onto substrates. Furthermore, due to their high metabolic activity and their role in aquatic food webs, microbial biofilms are also likely to influence contaminant fate in aquatic ecosystems. In this review, we provide (1) a critical overview of the analytical methods currently in use for detecting and quantifying metals and organic micropollutants in microbial biofilms attached to benthic substrata (rocks, sediments, leaf litter); (2) a review of the distribution of those contaminants within aquatic biofilms and the role of these benthic microbial communities in contaminant fate; (3) a set of future challenges concerning the role of biofilms in contaminant accumulation and trophic transfers in the aquatic food web. This literature review highlighted that most knowledge on the interaction between biofilm and contaminants is focused on contaminants dynamics in periphyton while technical limitations are still preventing a thorough estimation of contaminants accumulation in biofilms attached to leaf litter or sediments. In addition, microbial biofilms represent an important food resource in freshwater ecosystems, yet their role in dietary contaminant exposure has been neglected for a long time, and the importance of biofilms in trophic transfer of contaminants is still understudied.

Recycled low-density polyethylene composite to mitigate the environmental impacts generated from coal mining waste in Brazil.

Waste materials from coal mining and consumer products can pose significant risks to the environment. Residual coal deposits lead to the formation of acid drainage and release of contaminants, causing negative changes in soil and aquatic systems. Low density polyethylene (LDPE) polymers are an environmental concern due to their high useage, and slow degradation in the environment. In this study both waste materials were used to develop a composite to mitigate the environmental impacts of coal mining waste (CMW). The composite material was produced in different formulations (0-80 % wt CMW), and samples were tested for formation of acid drainage and release of contaminants. Chemical characterisation of the CMW and leachate of the composite materials was performed by X-ray fluorescence and atomic absorption spectrometry. Ecotoxicological effects in soil and water were investigated using standard tests with the earthworm, Eisenia fetida, the collembolan, Folsomia candida and the bacterium, Aliivibrio fischeri. Composites with 20 % wt LDPE showed a 50% increase in the pH value of the leachate compared to the CMW leachate. Iron, aluminium and sulfate concentrations were lower in leachates of the composite materials, and a reduction in the ecotoxicological impact on the tested organisms was observed. The hydrophobic nature of the composite's polymeric matrix as well its physical properties contributed to a better coating of the coal residue particles, blocking the contact with water and reducing the environmental risks of CMW. These results show that the production of composite material is a viable alternative route for treating coal and LDPE waste.

Refining uptake and depuration constants for fluoroalkyl chemicals in Chironomus riparius larvae on the basis of experimental results and modelling.

The aims of this study were to determine depuration rates for a range of per- and polyfluoroalkyl substances (PFASs) using Chironomus riparius, and to test a concentration-dependency hypothesis for the long-chain perfluorotridecanoic acid (PFTrDA) for this species. Midge larvae were exposed to field sediments collected downstream of a fluorotelomer plant, and to the same sediment spiked with PFTrDA. Elimination kinetics results indicated complete elimination of all PFASs by chironomids after 42h. These data were used to develop two PFTrDA bioaccumulation models accounting for chironomid growth and for compound concentration dependency or not. There was much better agreement between observed and simulated data under the concentration-dependency hypothesis than under the alternative one (passive diffusion). The PFTrDA uptake rate derived from the concentration-dependency model equaled 0.013 ± 0.008gocgwwh-1, and the depuration rate 0.032 ± 0.009h-1.

Field assessment of reproduction-related traits of chironomids using a newly developed emergence platform (E-Board).

Further progress in the development of reliable biomonitoring strategies requires to better link effects in aquatic ecological systems to ambient concentrations of chemical contaminants. Among existing tools, in situ bioassays using caging method represent an interesting way to achieve this challenge. However, elaboration of adapted exposure chambers and suitable operating procedures is still required, particularly to assess ecological relevant traits such as those related to the reproduction. In such context, we developed a new device (Emergence board - E-Board) which allows assessing in rivers the development of the Chironomus riparius species from the early fourth instar larvae to the adult stage. The system acts as a suspended matter trap floating in the subsurface of the water equipped of an emergence trap for catching adults. The system was tested in actual field conditions. Its easy handling allowed obtaining data which demonstrated its applicability for assessing the development of the chironomids. Moreover, by adapting energy-based models (DEB) specifically developed in the laboratory for the species C. riparius, we were able to predict the growth pattern and the emergence of chironomids in real environmental conditions. The E-Board represents thus a promising new in situ tool in perspective of evaluation of the quality of the ecosystems.

Bioaccumulation kinetics and effects of sediment-bound contaminants on chironomids in deep waters: new insights using a low-disturbance in situ system.

Effective assessment of sediment quality and associated risks requires further integration between laboratory-based studies and field observations. Tools for in situ exposure of laboratory-reared organisms are particularly suitable for this purpose. However, available tools suffer from technical drawbacks that limit their use in deep waters and their general ability to provide results linking laboratory and field observations. To overcome these limitations, we developed a new device (Sediment-Water Interface Study System - S-WISS l) for in situ exposure of fourth instar larvae of Chironomus riparius at depths up to 18 m. S-WISS 1 allows minimum alteration of the sediment natural structure (upon both deployment and recovery) and, in a way analogous to laboratory tests, repeated delivery of food doses to the in situ exposed organisms. Results obtained in situ with S-WISS1 were compared with standard laboratory tests and simple caging systems (large-mouthed bottles partly filled with sand) used as field controlks. A first series of experiments (48 h, no food addition) showed that element bioavailability to chironomids was higher in laboratory settings than on-site or in situ. A second series of experiments (96 h, food addition every 24 h) confirmed that sediments rather than overlying waters acted as the principal source of contaminants and that exposure to field sediments reduced chironomids' growth compared with on-site controls. S-WISSI represents a new family of in situ tools capable of combining the environmental realism associated with field monitoring with the controlled experimental conditions typical of laboratory testing.

The Genus Chaetogaster Baer, 1827 (Annelida, Clitellata) in Switzerland: A First Step toward Cataloguing Its Molecular Diversity and Description of New Species on a DNA Sequence Basis.

The genus Chaetogaster belongs to the subfamily Naidinae (Naididae); it includes mostly species of small size and is diverse and abundant in surface coarse sediments in streams. The aim of the present study is to initiate an inventory of lineages (=species) of Chaetogaster in Switzerland. We used 135 specimens collected at 6 sites in 4 streams of 4 cantons. We sequenced the cytochrome c oxidase (COI) gene from all specimens and ITS2 and rDNA 28S from all or several specimens of each lineage that was delimited using COI data, and preserved, for morphological identifications, the anterior part of almost all sequenced specimens. We were able to delimit, based on the calculation of genetic distances and analyses of single-locus data, one lineage for Chaetogaster diaphanus (Gruithuisen, 1828), three within Chaetogaster diastrophus (Gruithuisen, 1828), one for Chaetogaster langi Brestcher, 1896, one for Chaetogaster setosus Svetlov, 1925, and three unidentified Chaetogaster spp. Two lineages of Chaetogaster spp. could correspond to a new morphological group, but this should be confirmed in more specimens. We proposed a new identification key of the nominal species and described the three C. diastrophus lineages and two Chaetogaster spp. as new species. The prospects of the present work are to complete the data of the molecular diversity of this genus in Switzerland and to describe the newly found Chaetogaster species on a molecular/morphological basis.

Identification of polar organic chemicals in the aquatic foodweb: Combining high-resolution mass spectrometry and trend analysis.

Environmental risk assessment of chemical contaminants requires prioritizing of substances taken up by biota as it is a starting point for potential adverse effects. Although knowledge about the occurrence of known chemical pollutants in aquatic organisms has significantly improved during the last decade, there is still a poor understanding for a broad range of more polar compounds. To tackle this issue, we proposed an approach that identifies bioaccumulative and biomagnifiable polar chemicals using liquid chromatography coupled with electrospray ionization to high resolution tandem mass spectrometry (LC-HRMS/MS) and combine it with trend analysis using hierarchical clustering. As a proof-of-concept, this approach was implemented on various organisms and compartments (sediment, litter leaves, periphytic biofilm, invertebrates and fish) collected from a small urban river. HRMS/MS data measured via data-independent acquisition mode were retrospectively analysed using two analytical strategies: (1) retrospective target and (2) suspect/non-target screening. In the retrospective target analysis, 56 of 361 substances spanning a broad range of contaminant classes were detected (i.e. 26 in fish, 18 in macroinvertebrates, 28 in leaves, 29 in periphyton and 32 in sediments, with only 7 common to all compartments), among which 49 could be quantified using reference standards. The suspect screening approach based on two suspect lists (in-house, Norman SusDat) led to the confirmation of 5 compounds with standards (three xenobiotics at level 1 and two lipids at level 2) and tentative identification of seven industrial or natural chemicals at level 2 and 3 through a mass spectra library match. Overall, this proof-of-concept study provided a more comprehensive picture of the exposure of biota to emerging contaminants (i.e., the internal chemical exposome) and potential bioaccumulation or biomagnification of polar compounds along the trophic chain.

Incorporation of sediment- and soil-specific aspects in the Criteria for Reporting and Evaluating Ecotoxicity Data (CRED).

In environmental risk assessment either for registration purposes or for retrospective assessments of monitoring data, the hazard assessment is predominantly based on effect data from ecotoxicity studies. Most regulatory frameworks require studies used for risk assessment to be evaluated for reliability and relevance. Historically, the Klimisch methodology was used in many regulatory procedures where reliability needed to be evaluated. More recently, the Criteria for Reporting and Evaluating Ecotoxicity Data (CRED) have been developed for aquatic ecotoxicity studies, providing more detailed guidance on the evaluation and reporting of not only the reliability but also the relevance of a scientific study. Here, we discuss the application of the CRED methodology for assessing sediment and soil ecotoxicity studies, addressing important sediment- and soil-specific criteria that should be included as part of the CRED evaluation system. We also provide detailed recommendations for the design and reporting of sediment and soil toxicity studies that can be used by scientists and researchers wishing to contribute ecotoxicological data for effect assessments carried out within regulatory frameworks. Integr Environ Assess Manag 2024;20:2162-2174. © 2024 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Sediment quality assessment framework for per- and polyfluoroalkyl substances: Results from a preparatory study and regulatory implications.

A preparatory study was performed to develop a sediment quality and risk assessment strategy for Switzerland, addressing the following questions: the sediment fraction to be analyzed chemically (< 63 µm, or alternatively < 2 mm); the suitability of using perfluorooctanesulfonic acid (PFOS) as an indicator of per- and polyfluoroalkyl substances (PFAS) contamination in sediments; the availability of data for the derivation of sediment quality guidelines; and the suitability of normalization to total organic carbon (TOC). The results confirmed PFOS as a suitable indicator of PFAS contamination in sediments from small streams, being the most detected and on average with the highest concentrations among the analyzed PFAS. The fine fraction (< 63 µm) was more appropriate to screening for possible sources and studying the compound profiles at the study sites, but the analysis of the < 2 mm fraction and the normalization to a sample consisting of 100% of the < 63 µm fraction was, in principle, feasible for PFOS. Sediment quality guidelines for PFOS aiming to protect benthic invertebrates from generic adverse effects could be derived, but the available toxicity database is still too sparse. It was only possible to derive preliminary values. Sediment quality guidelines to protect wildlife and human health from secondary poisoning, using the equilibrium partitioning approach and simple trophic web models, were also derived. The use of food web models to derive sediment quality guidelines has not been validated, and the available database of Biota-Sediment Accumulation Factors and Trophic Magnification Factors for PFOS remains limited. However, it is still larger than the effect concentrations database based on sediment studies using benthic invertebrates. Normalization to TOC content in the field (0.2%-12.9%) was decisive in the quality assessment outcome. This is a widely accepted practice for hydrophobic organic contaminants, but its endorsement for PFOS would also benefit from validation. Integr Environ Assess Manag 2021;17:716-725. © 2021 SETAC.

Legacy and alternative halogenated flame retardants in Lake Geneva fish.

Legacy (i.e., polybrominated diphenyl ethers (PBDEs) and hexabromocyclododecane (HBCDD)) and alternative halogenated flame retardants (HFRs) were analyzed in 31 whole fish samples from Lake Geneva in 2018. Two fish species, namely, the burbot (Lota lota) and the roach (Rutilus rutilus), were selected, hypothetically representing different habitats, feeding behaviors, and different metabolic capacities. Roach (N = 20) and burbot (N = 11) displayed similar size and mass, but the latter species was overall leaner than the former. The sum of individual PBDE concentrations (0.54-9.86 ng g-1 wet weight (ww)) was similar in both species, but the respective molecular profiles suggested contrasted metabolic capacities. HBCDD sum of isomer concentrations ranged from non-detected to 3.477 ng g-1 (ww), also similar in both species. Both PBDEs and HBCDD levels were far below the threshold that indicates a risk to fish predators. Referring to previous surveys, which involved a wider range of species, PBDE concentrations have declined or are stable. HBCDD concentrations remained low, despite the PBDE ban, which could have fostered the consumption of other HFRs. The occurrence of alternative HFRs was also low for most compounds analyzed. Only dechloranes and decabromodiphenyl ethane (DBDPE) had detection rates above 50%. Dechloranes spanned a concentration range between 5 and 10 times the quantification limits (0.002 to 0.005 ng g-1 wet weight), lower than DBDPE (< 0.005 to 2.89 ng g-1 wet weight). Quality standards targeting biota are currently missing for these emerging chemicals.

Diversity, Functions and Antibiotic Resistance of Sediment Microbial Communities From Lake Geneva Are Driven by the Spatial Distribution of Anthropogenic Contamination.

Lake sediments are natural receptors for a wide range of anthropogenic contaminants including organic matter and toxicants such as trace metals, polycyclic aromatic hydrocarbons, polychlorinated biphenyls that accumulate over time. This contamination can impact benthic communities, including microorganisms which play a crucial role in biogeochemical cycling and food-webs. The present survey aimed at exploring whether anthropogenic contamination, at a large lake scale, can influence the diversity, structure and functions of microbial communities associated to surface sediment, as well as their genetic potential for resistance to metals and antibiotics. Changes in the characteristics of these communities were assessed in surface sediments collected in Lake Geneva from eight sampling sites in October 2017 and May 2018. These sampling sites were characterized by a large concentration range of metal and organic compound contamination. Variation between the two sampling periods were very limited for all sampling sites and measured microbial parameters. In contrast, spatial variations were observed, with two sites being distinct from each other, and from the other six sites. Benthic communities from the most contaminated sampling site (Vidy Bay, near the city of Lausanne) were characterized by the lowest bacterial and archaeal diversity, a distinct community composition, the highest abundance of antibiotic resistance genes and functional (respiration, denitrification, methanogenesis, phosphatase, and beta-glucosidase) activity levels. The second sampling site which is highly influenced by inputs from the Rhône River, exhibited low levels of diversity, a distinct community composition, high abundance of antibiotic resistance genes and the highest bacterial abundance. Overall, our results suggest that local anthropogenic contamination, including organic matter and toxicants, is a major driver of the diversity and functioning of sediment-microbial communities in Lake Geneva. This highlights the need to consider benthic microbial communities and a suite of complementary ecotoxicological endpoints for more effective environmental risk assessments of contaminants in lake sediments.

Chromium(VI) is more toxic than chromium(III) to freshwater algae: a paradigm to revise?

The behavior and toxicity of Cr(III) and Cr(VI) to the green algae Pseudokirchneriella subcapitata and Chlorella kessleri were studied in a standard culture medium (ISO medium) and, for P. subcapitata only, in ultrafiltered natural water enriched with all ISO components (modified ISO medium). In all solutions amended with Cr(III), initial chromium concentrations decreased by 60-90% over 72h (the duration of algal tests) indicating that protocols for testing poorly soluble substances are required to properly evaluate Cr(III) toxicity. After accounting for its behavior in test solutions, chromium(III) was 5-10 times more toxic than Cr(VI) in both media. For P. subcapitata, the average 72h EC50 of Cr(III) in ISO medium was 17.4+/-4.7 microg/L (n=9); lower than corresponding hardness-corrected Continuous Concentration Criteria of the US EPA and well within the range of Cr concentrations found in waters impacted by tannery discharges. These results follow from intrinsic chemical properties of Cr(III) in circumneutral solutions, so that the actual toxicity of Cr(III) to aquatic organisms may be generally underestimated.

The Foreign Oligochaete Species Quistadrilus multisetosus (Smith, 1900) in Lake Geneva: Morphological and Molecular Characterization and Environmental Influences on Its Distribution.

The presence of the oligochaete species Quistadrilus multisetosus (Smith, 1900) originating from North America has been mentioned for several decades in Europe, the Middle East and Russia. Its distribution and abundance in Europe is still unknown but it can be considered as potentially invasive. This species was recently discovered in Lake Geneva (Switzerland/France) and three other Swiss lakes. The aims of the present work are to report its repartition and abundance in Lake Geneva, to study its ecology and to determine its invasive potential in this lake. We also provide an identification key for correctly differentiating Q. multisetosus from the closely related species Spirosperma ferox Eisen, 1879 and Embolocephalus velutinus (Grube, 1879), and study the phylogenetic position of Q. multisetosus within several Tubificinae lineages based on the cytochrome c oxidase (COI) marker. Twenty-eight sites have been monitored since 2009 in Lake Geneva. In several sites, the COI sequence corresponding to this species was also searched for in sediment samples using high-throughput sequencing. In addition, we examined specimens collected in this lake before 2009 likely to belong to Q. multisetosus and to have been misidentified. We found that Q. multisetosus was only present in the lake downstream of a wastewater treatment plant and a combined sewer overflow in the Vidy Bay (near Lausanne) and at a site located nearby. These results confirmed the high tolerance of this species to organic matter pollution. Q. multisetosus was already present in this location in 1974 (misidentified as Spirosperma ferox), which suggests that Q. multisetosus has a limited capacity to disseminate in this lake. However, we recommend continuing monitoring its presence in Lake Geneva in the future, especially in the context of warming of waters that could contribute to the expansion of this species.

Towards simple tools to assess functional effects of contaminants on natural microbial and invertebrate sediment communities.

Surface sediments can accumulate contaminants that affect microorganisms and invertebrates and disturb benthic ecological functions. However, effects of contaminants on ecological functions supported by sediment communities are understudied. Here, we tested the relevance of two simple tools to assess the ecotoxicological effects of metal contamination on natural sediment communities using particulate organic matter breakdown and decomposition as a functional descriptor. To this aim, we performed a 21-day laboratory microcosm experiment to assess the individual and combined effects of Cu and As (nominal concentration of 40 mg kg-1 dw each) using the bait-lamina method (cellulose, bran flakes, and active coal in PVC strips) as well as artificial tablets (cellulose, bran flakes and active coal embedded in an agar matrix). Sediment toxicity was also evaluated using the standardized ostracod toxicity test. Both the bait-lamina and artificial tablet methods showed low effects of As on organic matter breakdown and decomposition but strong effects of Cu on this important ecological function. Both also showed that the presence of Cu and As in mixture in the sediment induced total inhibition of organic matter breakdown and decomposition. The ostracod toxicity test also showed high toxicity of Cu-spiked and Cu-plus-As-spiked sediments and low toxicity of As-spiked sediments. Besides confirming that artificial organic matter substrates are relevant and useful for assessing the functional effects of contaminants on sediment micro- and macro-organism communities, these results suggest that the proposed methods offer promising perspectives for developing tools for use in assessing functional ecotoxicology in the sediment compartment.

High-throughput DNA barcoding of oligochaetes for abundance-based indices to assess the biological quality of sediments in streams and lakes.

Aquatic oligochaete communities are valuable indicators of the biological quality of sediments in streams and lakes, but identification of specimens to the species level based on morphological features requires solid expertise in taxonomy and is possible only for a fraction of specimens present in a sample. The identification of aquatic oligochaetes using DNA barcodes would facilitate their use in biomonitoring and allow a wider use of this taxonomic group for ecological diagnoses. Previous approaches based on DNA metabarcoding of samples composed of total sediments or pools of specimens have been proposed for assessing the biological quality of ecosystems, but such methods do not provide precise information on species abundance, which limits the value of resulting ecological diagnoses. Here, we tested how a DNA barcoding approach based on high-throughput sequencing of sorted and genetically tagged specimens performed to assess oligochaete species diversity and abundance and the biological quality of sediments in streams and lakes. We applied both molecular and morphological approaches at 13 sites in Swiss streams and at 7 sites in Lake Geneva. We genetically identified 33 or 66 specimens per site. For both approaches, we used the same index calculations. We found that the ecological diagnoses derived from the genetic approach matched well with those of the morphological approach and that the genetic identification of only 33 specimens per site provided enough ecological information for correctly estimating the biological quality of sediments in streams and lakes.

Retrospective screening of high-resolution mass spectrometry archived digital samples can improve environmental risk assessment of emerging contaminants: A case study on antifungal azoles.

Environmental risk assessment associated with aquatic and terrestrial contamination is mostly based on predicted or measured environmental concentrations of a limited list of chemicals in a restricted number of environmental compartments. High resolution mass spectrometry (HRMS) can provide a more comprehensive picture of exposure to harmful chemicals, particularly through the retrospective analysis of digitally stored HRMS data. Using this methodology, our study characterized the contamination of various environmental compartments including 154 surface water, 46 urban effluent, 67 sediment, 15 soil, 34 groundwater, 24 biofilm, 41 gammarid and 49 fish samples at 95 sites widely distributed over the Swiss Plateau. As a proof-of-concept, we focused our investigation on antifungal azoles, a class of chemicals of emerging concern due to their endocrine disrupting effects on aquatic organisms and humans. Our results demonstrated the occurrence of antifungal azoles and some of their (bio)transformation products in all the analyzed compartments (0.1-100 ng/L or ng/g d.w.). Comparison of actual and predicted concentrations showed the partial suitability of level 1 fugacity modelling in predicting the exposure to azoles. Risk quotient calculations additionally revealed risk of exposure especially if some of the investigated rivers and streams are used for drinking water production. The case study clearly shows that the retrospective analysis of HRMS/MS data can improve the current knowledge on exposure and the related risks to chemicals of emerging concern and can be effectively employed in the future for such purposes.

Investigation of the spatial variability of poly- and perfluoroalkyl substance trophic magnification in selected riverine ecosystems.

The occurrence at different trophic levels of 17 poly- and perfluoroalkyl substances (PFASs), including perfluoroalkyl acids (PFAAs) and some of their precursors (e.g., perfluoroalkane sulfonamides, 6:2 fluorotelomer sulfonate (6:2 FTSA)), was investigated in riverine freshwater food webs in Southeastern France. Two fish species (Barbus barbus and Squalius cephalus) and various invertebrate taxa were collected in five rivers to assess the spatial variability of trophic magnification factors (TMFs). Particular attention was devoted to sample and data processing to minimize potential biases associated with the TMF determination. Fish were significantly more contaminated than invertebrates (ΣPFAS = 7-1811 vs. 0.9-213 ng g-1 wet weight (ww)). Those from the Rhône River presented significantly higher levels due to high concentrations of perfluoroundecanoic acid (406 ng g-1 ww) and perfluorotridecanoic acid (566 ng g-1 ww) ascribed to an industrial point source. Perfluorooctane sulfonate (PFOS) was dominant at the other sites (concentration range = 3.6-134 ng g-1 ww). Two linear regression models were compared (i.e., Kendall regression vs. Generalized Linear Mixed-Effect Model, GLMM). Results showed that TMFs calculated using the non-weighted Kendall regression were higher than those obtained using the GLMM approach. GLMM-based TMFs were consistently >1 for C9-C14 perfluorocarboxylic acids (PFCAs), PFOS and perfluorodecane sulfonate (PFDS), indicating their apparent biomagnification in the investigated food webs. Comparatively, 6:2 FTSA and N-ethylperfluorooctane sulfonamidoacetic acid (N-EtFOSAA) were less often detected and were not significantly biomagnified, probably because of metabolization. TMF estimates were generally consistent across sites although some PFASs (in particular C9, C10 and C13 PFCAs) displayed higher variability, due to a unique extreme value that may have resulted from the contribution of unattributed precursor biotransformation.

Simultaneous preservation of the DNA quality, the community composition and the density of freshwater oligochaetes for the development of genetically based biological indices.

INTRODUCTION: Oligochaetes are recognized as valuable bioindicators of sediment quality in streams and lakes. The development of an oligochaete index based on the identification of specimens using DNA barcodes requires a method for simultaneously preserving the DNA quality and information on the specimen density and oligochaete community composition. Absolute ethanol optimally preserves DNA but fixation of freshwater oligochaetes with this medium can cause disintegration and fragmentation of specimens. Here, we investigated the possibility to preserve oligochaete specimens in low-pH formalin and in neutral buffered formalin for up to four weeks before genetic analyses and tested if the addition of absolute ethanol to formalin-fixed oligochaetes resulted in a loss of specimens and/or species. METHODS: We performed guanidine extraction and polymerase chain reaction (PCR) amplification/sequencing of a fragment of the cytochrome c oxidase I (COI) gene on tissue fragments preserved in low-pH formalin for up to 3 weeks and in neutral buffered formalin for up to 4 weeks. In addition, we compared the density and taxonomic composition of formalin-fixed oligochaetes of several sieved sediment samples before and after the addition of absolute ethanol. RESULTS: The COI fragment of all oligochaete specimens preserved in neutral buffered formalin for up to 28 days was successfully amplified by PCR and obtained sequences were complete and of high quality. The amplification success rate for low-pH formalin fixed specimens declined after 7 days of storage. The addition of absolute ethanol to formalin-fixed oligochaete communities did not alter density or diversity estimates. DISCUSSION: Our results indicate that sediment samples can be stored in neutral buffered formalin for up to 4 weeks and the sieved material can then be transferred to absolute ethanol, without affecting DNA quality, density and community composition of oligochaetes. Based on these results, a protocol for preserving freshwater oligochaetes, describing all the steps from collection of sediments to preservation of the biological material in absolute ethanol, is proposed. This method of fixation/preservation is of relevance for establishing DNA barcode reference databases, inventories of genetic diversity and developing genetically based biological indices.