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The global biodiversity crisis emphasizes our need to understand how different stressors (climatic, chemical, parasitic, etc.) interact and affect biological communities. We provide a comprehensive meta‑analysis investigating joint effects of chemical and parasitic stressors for 1064 chemical‑parasitic combinations using the Multiplicative model on mortality of arthropods. We tested both features of the experimental setup (control mortality, stressor effect level) and the chemical mode of action, host and parasite phylogeny, and parasite‑host interaction traits as explanatory factors for deviations from the reference model. Synergistic interactions, defined as higher mortality than predicted, were significantly more frequent than no interactions or antagony. Experimental setup significantly affected the results, with studies reporting high (> 10%) control mortality or using low stressor effects (< 20%) being more synergistic. Chemical mode of action played a significant role for synergy, but there was no effects of host and parasite phylogeny, or parasite‑host interaction traits. The finding that experimental design played a greater role in finding synergy than biological factors, emphasize the need to standardize the design of mixed stressor studies across scientific disciplines. In addition, combinations testing more biological traits e.g. avoidance, coping, and repair processes are needed to test biology‑based hypotheses for synergistic interactions., The global biodiversity crisis emphasizes our need to understand how different stressors (climatic, chemical, parasitic, etc.) interact and affect biological communities. We provide a comprehensive meta-analysis investigating joint effects of chemical and parasitic stressors for 1064 chemical-parasitic combinations using the Multiplicative model on mortality of arthropods. We tested both features of the experimental setup (control mortality, stressor effect level) and the chemical mode of action, host and parasite phylogeny, and parasite-host interaction traits as explanatory factors for deviations from the reference model. Synergistic interactions, defined as higher mortality than predicted, were significantly more frequent than no interactions or antagony. Experimental setup significantly affected the results, with studies reporting high (> 10%) control mortality or using low stressor effects (< 20%) being more synergistic. Chemical mode of action played a significant role for synergy, but there was no effects of host and parasite phylogeny, or parasite-host interaction traits. The finding that experimental design played a greater role in finding synergy than biological factors, emphasize the need to standardize the design of mixed stressor studies across scientific disciplines. In addition, combinations testing more biological traits e.g. avoidance, coping, and repair processes are needed to test biology-based hypotheses for synergistic interactions.

Abstract: Plants represent uncommon targets to evaluate pharmaceuticals toxicity. In this work, Lemna minor was employed as a plant model to determine the toxicity of selected pharmaceuticals, and to assay if such toxicity could be predicted by QSAR models based on green algae. Among eight compounds, measurable toxicity was determined for ketoprofen (EC50 = 11.8 ± 1.9 mg/L), fluoxetine (EC50 = 27.0 ± 8.7 mg/L) and clindamycin 2-phosphate (EC50 = 57.7 ± 1.7 mg/L). Even though a correlation of r2 = 0.87 was observed between experimental toxicity towards algae and L. minor, QSAR estimations based on algae data poorly predicted the toxicity of pharmaceuticals on the plant. More experimental data for L. minor are necessary to determine the applicability of these predictions; nonetheless, these results remark the importance of measuring experimental ecotoxicological parameters for individual taxa. The toxicity of pharmaceutical binary mixtures (ketoprofen, fluoxetine and clindamycin) revealed in some cases deviations from the concentration addition model; nonetheless these deviations were small, thus the interactions are unlikely to be of severe biological significance. Moreover, the EC50 concentrations determined for these pharmaceuticals are significantly higher than those detected in the environment, suggesting that acute effects on L. minor would not take place at ecosystem level.

Background: Triterpenoid saponins are bioactive compounds synthesised by plants in defence against insects or fungi. With a demand for new, sustainable, plant protection products saponins are under consideration as green insecticides. This study tries to identify the acute toxic effects of 10 saponin structures towards different non-target organisms and relate the effects to their chemical structure. Results: a-Hederin showed effects towards all test organisms, such as the aquatic crustacean Daphnia magna, the annelid worm Enchytraeus crypticus, the green algae Raphidocelis subcapitata and the yeast Saccaromyces cerevisiae. Highest toxicity was observed for a-Hederin (EC50 8.6 mg L-1) and Hederacolchisid-A1 (EC50 2.03 mg L-1) towards the annelid worm E. crypticus. The aquatic crustacean Daphnia magna was sensitive to Hederacolchisid- A1, a-Hederin, Arjunetin and Quillaja-mix with EC50 values from 12.52 mg L-1 - 16.90 mg L-1. S. cerevisiae was tested against a-Hederin and Quillaja-extract only and was more susceptible to a-Hederin (EC50 6.77 mg L-1 and 243.20 mg L-1). R. subcapitata showed minor effects when exposed to a-Hederin (EC50 42.99 mg L-1) and Madecassosid (267.65 mg L-1). This indicates that the algae, are the least sensitive species when exposed to saponins. Conclusion: Saponin toxicity depends on the interactions with sterols embedded in cell membranes. Our results indicate that a glycosylation at their C3 position together with a free carboxylic acid group at the C28 position allow triterpenoid saponins to interact with cholesterol and ergosterol and to a weaker extent with the plant sterol stigmasterol. Arjunetin and Quillaja-extract might be capable of interfering with chitin.

Applying organic wastes such as manure or sewage sludge on agricultural fields improves soil physical structure and fertility, while giving purpose to otherwise disposed and burned waste products. Yet, xenobiotics, and especially pharmaceuticals and personal care products, may be present in these wastes and thereby enter the soil ecosystem. To investigate their occurrence and their potential effect on soil fauna, the CRUCIAL experimental fields, Denmark, were used. Here we combine, for the first time, a chemical wide-scope suspect screening analysis with a study of soil nematode faunal response on soils, which were fertilized with cattle manure, sewage sludge, human urine, and mineral fertilizers over 14 years. The suspect screening analysis was performed with a nanoflow ultra-high performance liquid chromatography coupled with high-resolution tandem mass spectrometry. The abundance of soil indigenous nematodes was determined in each treatment at two time-points, followed by laboratory tests with the nematode species Caenorhabditis elegans to assess its reproduction capacity in the soils. A total of 12 pharmaceuticals and personal care products were identified as being up- or down-concentrated in the different treatments compared to controls (mineral fertilizers): 8 and 2 were found to be up-concentrated in sludge- and urine-treated soils, respectively. Our results, however, showed a significantly greater abundance of nematodes in soils amended with sludge and manure, while C. elegans had the lowest reproduction capacity in the controls. The reproduction capacity tests did not reveal any chronic toxicity. A principal component analysis showed a clear correlation between nematode abundance and soil organic matter, water content, nutrient contents, and porosity. Overall, urban and animal wastes enhanced soil quality. Pharmaceuticals were detected, but adverse effects on the measured endpoints could not be demonstrated. Although further investigations are needed to

Man-made chemicals are a significant contributor to the ongoing deterioration of numerous ecosystems. Currently, risk assessment of these chemicals is based on observations in a single generation of animals, despite potential adverse intergenerational effects. Here, we investigate the effect of the fungicide prochloraz across three generations of Daphnia magna. We studied both the effects of continuous exposure over all generations and the effects of first-generation (F0) exposure on two subsequent generations. Effects at different levels of biological organization from genome-wide gene expression, whole organism metabolite levels, CYP enzyme activity and key phenotypic effects, such as reproduction, were monitored. Acclimation to prochloraz was found after continuous exposure. Following F0-exposure, embryonically exposed F1-offspring showed no significant effects. However, in the potentially germline exposed F2 animals, several parameters differed significantly from controls. A direct association between these F2 effects and the toxic mode of action of prochloraz was found, showing that chemicals can be harmful not only to the directly exposed generation, but also to prenatally exposed generations and in that way effects may even appear to skip a generation. This implies that current risk assessment practices are neglecting an important aspect of toxicity, such as delayed effects across generations due to a time gap between chemical exposure and emergence of effects.

Hydrolysis catalyzed by general esterases (GEs) is the most efficient route for hydrolyzation of pyrethroid insecticides. Organophosphate (OP) and carbamate (CB) insecticides are known to inhibit GEs in addition to acetylcholinesterase (AChE), which is their main target. We hypothesize that synergies can be induced by OPs and CBs when mixed with pyrethroids, due to their inhibition of GE-dependent detoxification of pyrethroids. To test this hypothesis, we conducted mixture toxicity experiments with Daphnia magna using alpha-cypermethrin (alpha-cyp) in combination with the noninsecticidal OP tetraisopropyl pyrophosphoramide (iso-OMPA) and five AChE inhibitors diazinon, chlorpyrifos, chlorfenviphos, parathion, and aldicarb. In addition, the in vivo GE activity inhibition was measured for all compounds. Up to 10-fold synergy was found between alpha-cyp and iso-OMPA, and the degree of synergy correlated linearly with the inhibition of the GE activity. No synergy, however, was found in any of the insecticide mixtures nor was the GE activity inhibited within the nonlethal concentration range tested. It was concluded that the effect of the insecticides on AChE occurred at lower concentrations than their effect on GEs, making the daphnids become immobilized before any synergistic effects on mortality could be observed. The implications of the findings are discussed from a risk assessment perspective.

Acetylcholinesterase (AChE) and general-esterase (GE) activities are important to understand detoxification processes of xenobiotics. The assays to quantify them have employed different substrates, inhibitors, types of experiments (in vitro and in vivo) and model organisms. The aim of this work was to give a systematic overview of the effect of the above factors on the outcome of AChE and GE activity measurements. We showed that AChE activity could be measured with the substrate acetylthiocholine iodide (AChI) but not with acetylcholine bromide (AChB) and only in in vitro assays. For GE activity, Michaelis-Menten kinetics differed between the substrates 4-methylumbellifery butyrate (4-MUB) and 1-naphtyl acetate (1-NA) in the measurements of in vitro activity, but their inhibition curves and IC50 values for the general inhibitor tetraisopropyl pyrophosphoramide (iso-OMPA) were similar, confirming that both substrates targeted the same group of enzymes. The GE substrate 4-MUB was applicable both in vitro and in vivo, while 1-NA was only applicable in vitro due to its high acute toxicity. When comparing the zooplankton crustacean Daphnia magna and the sediment dwelling Chironomus riparius, the latter had a four-fold higher maximal AChE activity (Vmax) and a higher susceptibility to the AChE inhibitor BW284c51 (four-fold lower 50% inhibitory concentration, IC50), but a lower maximal GE activity and lower susceptibility to iso-OMPA (higher IC50), indicating significant species differences between in C. riparius and D. magna. We conclude that both choice of substrate and exposure method matters for the outcome of esterase assays and that esterase compositions between species may vary significantly.

Pyrethroid insecticides are known to be highly toxic to most aquatic nontarget organisms, but little is known about the mechanisms causing some species to be highly sensitive while others are hardly affected by the pyrethroids. The aim of the present study was to measure the sensitivity (EC50-values) of 10 aquatic invertebrates toward a 24 h pulse of the pyrethroid cypermethrin and subsequently test if the difference in sensitivity could be explained by measured morphological and physiological traits and modeled toxicokinetic (TK) and toxicodynamic (TD) parameters. Large differences were observed for the measured uptake and elimination kinetics, with bioconcentration factors ( BCFs) ranging from 53 to 2337 at the end of the exposure. Similarly, large differences were observed for the TDs, and EC50-values after 168 h varied 120-fold. Modeling the whole organism cypermethrin concentrations indicated compartmentation into a sorbed fraction and two internal fractions: a bioavailable and nonbioavailable internal fraction. Strong correlations between surface/volume area and the TK parameters (sorption and uptake rate constants and the resulting BCF) were found, but none of the TK parameters correlated with sensitivity. The only parameter consistently correlating with sensitivity across all species was the killing rate constant of the GUTS-RED-SD model (the reduced general unified threshold models of survival assuming stochastic death), indicating that sensitivity toward cypermethrin is more related to the TD parameters than to TK parameters.

Contains fulltext : 204310.pdf (publisher's version ) (Open Access)

In April 2017, the Organic Business Development Team released a report with 25 recommendations for the Minister of Environment and Food (Det økologiske erhvervsteam 2017). Among these was a recommendation that organic farmers should have opportunities for utilizing nutrients from treated domestic wastewater for nutrient recycling. A prerequisite for future use of nutrients from treated wastewater is, that quality requirements are met and that application can be explained to (and accepted by) consumers. In partial fulfilment of this, the business team identified a need for a scientific overview of the risks of using nutrients from treated municipal wastewater in relation to other authorized fertilizer sources – e.g. conventional animal manures. Thus, it was assumed that a comparative approach to assess potential risk of using sewage sludge and conventional manures, could usefully inform decision makers in the future regulation of organic farming systems. Dependent on the result of the scientific investigation, the Organic Business Development Team foresaw that Denmark could chose to work to expand Annex 1 of the EU Ecology Regulation, to allow the organic farmers to use nutrients from municipal wastewater or other acceptable derived sludge products. Mobilization of support for this should be done by the Ministry of Environment and Food in collaboration with the Organic Farming Industry. Thus, based on available literature, this report aims at creating an overview of the environmental and human risks associated with application of pig and cattle slurry as well as sewage sludge to agricultural soils. The risk evaluation was performed for the following compound groups: Metals, Chlorophenyls, Dioxins, Furans, Halogenated aliphatic and aromatic hydrocarbons (HAH), Linear alkylbenzenesulfonates (LAS), Polyaromatic hydrocarbons (PAH), Polybrominated diphenyl ethers (PBDE), Polychlorinated biphenyls (PCB), Poly- and perfluorinated alkylated substances (PFAS), Phenols, Phosph

Saponins form a group of plant-produced glycosides with potential as biopesticide ingredients. The environmental fate of saponins has never been fully investigated. In the present study, we use QS-18, a specific saponin from Quillaja saponaria as an example, to quantify hydrolysis under different conditions of pH, temperature and water chemical composition. Saponin hydrolysis in buffer solutions was base-catalyzed and followed first-order kinetics. Thus, hydrolysis was slow at pH 5.1 with a half-life of 330 ± 220 d (26 °C), which increases to 0.06 ± 0.01 d at pH 10.0. Hydrolysis rates were highly sensitive to temperature with an activation energy of 56.9 ± 14.2 kJ mol-1 at pH 7.2. In strong contrast, hydrolysis in lake waters (pH 6.4-8.2) produced different patterns with a fast initial dissipation of 25 to 60% of the added saponin within the first five hours, followed by an extremely slow reaction with 25 to 75% unreacted saponin left after reaction times longer than 120 h. The fast dissipation followed by slow hydrolysis in lake water was hypothesized to be attributed to sorption and/or flocculation of saponins by inorganic nanoparticles and/or solutes in the lake water followed by inactivation of hydrolysis due to the sorption/flocculation. The present study demonstrates that saponins may hydrolyze slowly under acidic and cold conditions. In addition, it demonstrates that dissipation kinetics in natural waters may deviate substantially from the kinetics predicted based on laboratory experiments with "clean" buffered solutions. This emphasizes the need for a deeper understanding of the processes affecting the dissipation kinetics of potential toxins under natural conditions, as fate models based on laboratory derived kinetic data may be seriously flawed.

The toxicity of the antifouling compounds diuron, irgarol, zinc pyrithione (ZnPT), copper pyrithione (CuPT) and copper was tested on the three marine microalgae Tisochrysis lutea, Skeletonema marinoi and Tetraselmis suecica. Toxicity tests based on the inhibition of growth rate after 96-h exposure were run using microplates. Chemical analyses were performed to validate the exposure concentrations and the stability of the compounds under test conditions. Single chemicals exhibited varying toxicity depending on the species, irgarol being the most toxic chemical and Cu the least toxic. Selected binary mixtures were tested and the resulting interactions were analyzed using two distinct concentration-response surface models: one using the concentration addition (CA) model as reference and two deviating isobole models implemented in R software; the other implementing concentration-response surface models in Excel®, using both CA and independent action (IA) models as reference and three deviating models. Most mixtures of chemicals sharing the same mode of action (MoA) were correctly predicted by the CA model. For mixtures of dissimilarly acting chemicals, neither of the reference models provided better predictions than the other. Mixture of ZnPT together with Cu induced a strong synergistic effect on T. suecica while strong antagonism was observed on the two other species. The synergy was due to the transchelation of ZnPT into CuPT in the presence of Cu, CuPT being 14-fold more toxic than ZnPT for this species. The two modelling approaches are compared and the differences observed among the interaction patterns resulting from the mixtures are discussed.

Following a request from EFSA, the Panel on Plant Protection Products and their Residues (PPR) developed an opinion on the state of the art of Toxicokinetic/Toxicodynamic (TKTD) models and their use in prospective environmental risk assessment (ERA) for pesticides and aquatic organisms. TKTD models are species- and compound-specific and can be used to predict (sub)lethal effects of pesticides under untested (time-variable) exposure conditions. Three different types of TKTD models are described, viz., (i) the ‘General Unified Threshold models of Survival’ (GUTS), (ii) those based on the Dynamic Energy Budget theory (DEBtox models), and (iii) models for primary producers. All these TKTD models follow the principle that the processes influencing internal exposure of an organism, (TK), are separated from the processes that lead to damage and effects/mortality (TD). GUTS models can be used to predict survival rate under untested exposure conditions. DEBtox models explore the effects on growth and reproduction of toxicants over time, even over the entire life cycle. TKTD model for primary producers and pesticides have been developed for algae, Lemna and Myriophyllum. For all TKTD model calibration, both toxicity data on standard test species and/or additional species can be used. For validation, substance and species-specific data sets from independent refined-exposure experiments are required. Based on the current state of the art (e.g. lack of documented and evaluated examples), the DEBtox modelling approach is currently limited to research applications. However, its great potential for future use in prospective ERA for pesticides is recognised. The GUTS model and the Lemna model are considered ready to be used in risk assessment.