How to analyse and account for interactions in mixture toxicity with toxicokinetic-toxicodynamic models.

The assessment of chemical mixture toxicity is one of the major challenges in ecotoxicology. Chemicals can interact, leading to more or less effects than expected, commonly named synergism and antagonism respectively. The classic ad hoc approach for the assessment of mixture effects is based on dose-response curves at a single time point, and is limited to identifying a mixture interaction but cannot provide predictions for untested exposure durations, nor for scenarios where exposure varies in time. We here propose a new approach using toxicokinetic-toxicodynamic modelling: The General Unified Threshold model of Survival (GUTS) framework, recently extended for mixture toxicity assessment. We designed a dedicated mechanistic interaction module coupled with the GUTS mixture model to i) identify interactions, ii) test hypotheses to identify which chemical is likely responsible for the interaction, and finally iii) simulate and predict the effect of synergistic and antagonistic mixtures. We tested the modelling approach experimentally with two species (Enchytraeus crypticus and Mamestra brassicae) exposed to different potentially synergistic mixtures (composed of: prochloraz, imidacloprid, cypermethrin, azoxystrobin, chlorothalonil, and chlorpyrifos). Furthermore, we also tested the model with previously published experimental data on two other species (Bombus terrestris and Daphnia magna) exposed to pesticide mixtures (clothianidin, propiconazole, dimethoate, imidacloprid and thiacloprid) found to be synergistic or antagonistic with the classic approach. The results showed an accurate simulation of synergistic and antagonistic effects for the different tested species and mixtures. This modelling approach can identify interactions accounting for the entire time of exposure, and not only at one time point as in the classic approach, and provides predictions of the mixture effect for untested mixture exposure scenarios, including those with time-variable mixture composition.

Environmental risks to freshwater organisms from the mycotoxins deoxynivalenol and zearalenone using Species Sensitivity Distributions.

In this study, laboratory experiments have addressed the acute toxicity of two common mycotoxins, deoxynivalenol (DON) and zearalenone (ZON), in a range of freshwater organisms (including rotifers Brachionus calyciflorus, insects Chironomus riparius (larvae), crustaceans Daphnia pulex and Thamnocephalus platyurus, cnidarians Hydra vulgaris, molluscs Lymnaea stagnalis (embryos) and Protozoa Tetrahymena thermophila). Acute EC50 values highlight crustaceans as the most sensitive organisms to DON, with T. platyurus having a 24 h EC50 of 0.14 and D. magna having a 48 h EC50 of 0.13 mg DON/L. During exposures to ZON, H. vulgaris and L. stagnalis embryos showed the highest sensitivity; mortality EC50 values were 1.1 (96 h) and 0.42 mg ZON/L (7 d), respectively. Combining these novel invertebrate toxicity results, along with recent published data for freshwater plant and fish toxicity for analysis of Species Sensitivity Distributions, provides freshwater HC5 values of 5.2 µg DON/L and 43 µg ZON/L, respectively. Using highest reported environmental concentrations and following REACH guidelines, risk ratios calculated here show the risk of ZON to freshwater organisms is low. In contrast, DON may periodically because for concern in streams subject to high agricultural run-off, likely during certain times of year where cereal crops are susceptible to higher fungal infections rates and may pose increased risks due to climate change.

Impacts of the mycotoxin zearalenone on growth and photosynthetic responses in laboratory populations of freshwater macrophytes (Lemna minor) and microalgae (Pseudokirchneriella subcapitata).

Mycotoxins are an important class of chemicals of emerging concern, recently detected in aquatic environments, potentially reflecting the influence of fungicide resistance and climatic factors on fungal diseases in agricultural crops. Zearalenone (ZON) is a mycotoxin formed by Fusarium spp. and is known for its biological activity in animal tissues; both in vitro and in vivo. ZON has been reported in US and Polish surface waters at 0.7 - 96 ng/L, with agricultural run-off and wastewater treatment plants being the likely sources of mycotoxins. As some mycotoxins can induce phytotoxicity, laboratory studies were conducted to evaluate the toxicity of ZON (as measured concentrations) to freshwater algae (Pseudokirchneriella subcapitata) and macrophytes (Lemna minor) following OECD test guidelines 201 and 221, respectively. Zinc sulphate was used as a positive control. In the OECD 201 algal static study (72 h at 24 ±â€¯1 °C), exposure to ZON gave average specific growth rate (cell density) EC50 and yield (cell density) EC50 values of > 3.1 and 0.92 (0.74 - 1.8) mg/L, respectively. ZON was less toxic in the OECD 221 static study and after 7 d at 24 ±â€¯1 °C. L. minor growth was significantly reduced based on frond number and frond area at 11.4 mg ZON/L, showing a higher tolerance than reported for other mycotoxins with Lemna spp. Chlorophyll fluorescence parameters were used as biomarkers of impacts on photosystem II efficiency, with no effect seen in algae but, with responses being observed in L. minor between 5.2 - 14.4 mg ZON/L. ZON toxicity seen here is not of immediate concern in context with environmental levels, but this study highlights that other freshwater organisms including algae are more sensitive to mycotoxins than Lemna sp., the only current source of toxicity data for freshwater plants.