How global warming changes the toxicity of contaminants is a research priority at the intersection of global change biology and ecotoxicology. While many pesticides are more toxic at higher temperatures this is not always detected. We studied whether deviations from this general pattern can be explained by concentration-dependent interaction effects and by testing the interaction against the inappropriate null model. We exposed larvae of the mosquito Culex pipiens to three concentrations of the pesticide chlorpyrifos (absence, low and high) in the absence and presence of 4°C warming. Both the low and high chlorpyrifos concentration were lethal and generated negative sublethal effects: activity of acetylcholinesterase (AChE) and total fat content decreased, and oxidative damage to lipids increased, yet growth rate increased. Warming was slightly lethal, yet had positive sublethal effects: growth rate, total fat content and metabolic rate increased, and oxidative damage decreased. For four out of seven response variables the independent action model identified the expected synergistic interaction between chlorpyrifos and warming. Notably, for three variables (survival, AChE and fat content) this was strongly dependent on the chlorpyrifos concentration, and for two of these (AChE and fat content) not associated with a significant interaction in the general(ized) linear models. For survival and fat content, warming only potentiated chlorpyrifos (CPF) toxicity at the low CPF concentration, while the opposite was true for AChE. Our results highlight that taking into account concentration-dependence and appropriate null model testing is crucial to improve our understanding of the toxicity of contaminants in a warming world.

The widespread evidence that global warming can increase species sensitivities to chemical toxicants and vice versa, and the recent insight that thermal evolution may mitigate these effects is crucial to predict the future impact of toxicants in a warming world. Nevertheless, a major component of global warming, the predicted increase in daily temperature fluctuations (DTFs), has been ignored at the interface of evolutionary ecotoxicology and global change biology. We studied whether 4°C warming and an 5°C DTF increase (to 10°C DTF) magnified the negative impact of the insecticide chlorpyrifos (CPF) in larvae of low-and high-latitude populations of the damselfly Ischnura elegans. While 4°C warming only increased CPF-induced mortality in high-latitude larvae, the high (10°C) DTF increased CPF-induced larval mortality at both latitudes. CPF reduced the heat tolerance, however, this was buffered by latitude-specific thermal adaptation to both mean temperature and DTF. Integrating our results in a space-for-time substitution indicated that gradual thermal evolution in high-latitude larvae may offset the negative effects of CPF on heat tolerance under warming, unless the expected DTF increase is taken into account. Our results highlight the crucial importance of jointly integrating DTFs and thermal evolution to improve risk assessment of toxicants under global warming.

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The exposure order may strongly affect the impact of stressors, yet is largely ignored for the frequently occurring combinations of toxicants with natural stressors. We tested how exposure order shaped the interactive effects of serial exposure to the pesticide chlorpyrifos and to a heat spike in the larvae of the mosquito Culex pipiens. Notably, the chlorpyrifos-induced mortality was much more magnified by the heat spike and a synergism was already detected at the low concentration when exposure to chlorpyrifos followed the heat spike. This suggests that the preceding heat spike weakened the larvae as reflected in their lower net energy budget, moreover the chlorpyrifos-induced inhibition of its target enzyme (acetylcholinesterase) was only magnified by the heat spike when it was the first stressor. Also the chlorpyrifos-induced reduction in heat tolerance was stronger when the pesticide pulse followed the heat spike, and was buffered by the heat spike when this was the second stressor. Our results provide the first evidence that the exposure order can strongly change the magnifying effect of an important climate change factor on the toxicity of a pesticide. This highlights the importance of exposure order in ecological risk assessment of toxicants under realistic combinations with natural stressors.

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Pesticide mixtures are increasingly used to fight pest species that developed resistance to pesticides. To assess the pesticide control efficiency and to reduce ecological damage to nontarget species, it is important to quantify the effect of these mixtures and compare them with the effect of their single pesticides on pest species, non-target species and their predator-prey interactions. We studied the effects of the chemical pesticide chlorpyrifos (CPF), the biopesticide Bacillus thuringiensis israelensis (Bti) and their mixture both on the direct mortality and on the mortality by predation. We focused on larvae of a CPF-resistant and a non-resistant strain of the vector mosquito Culex quinquefasciatus and its predator, the pygmy backswimmer Plea minutissima. In the CPF-Bti mixture, both pesticides interacted antagonistically for direct mortality. Exposure to the mixture caused equal direct mortality and equal mortality by predation in both strains. As expected, exposure to CPF resulted in less direct mortality and less mortality by predation in the CPF-resistant mosquito strain compared to the non-resistant strain. Notably, Bti caused a higher mortality in the mosquito larvae of the CPF-resistant strain compared to the non-resistant strain. Furthermore, the predator killed more mosquito larvae of the resistant strain compared to the non-resistant strain when exposed before to Bti alone. These observations identify a novel cost of resistance to a chemical pesticide in terms of increased vulnerability to a biopesticide.

Aquatic organisms rely on microbial symbionts for coping with various challenges they encounter during stress and for defending themselves against predators, pathogens and parasites. Microbial symbionts are also often indispensable for the host's development or life cycle completion. Many aquatic ecosystems are currently under pressure due to diverse human activities that have a profound impact on ecosystem functioning. These human activities are also ex pected to alter interactions between aquatic hosts and their associated microbes. This can directly impact the host's health and -given the importance and widespread occurrence of microbial symbiosis in aquatic systems -the ecosystem at large. In this review, we provide an overview of the importance of microbial symbionts for aquatic organisms, and we consider how the beneficial services provided by microbial symbionts can be affected by human activities. The scarcity of available studies that assess the functional consequences of human impacts on aquatic microbial symbioses shows that our knowledge on this topic is currently limited, making it difficult to draw general conclusions and predict future changes in microbial symbiont−host relationships in a changing world. To address this important knowledge gap, we provide an overview of ap proaches that can be used to assess the impact of human disturbances on the functioning of aquatic microbial symbioses.