The effect of warming on pesticide toxicity is reversed between developmental stages in the mosquito Culex pipiens.

A better understanding of interactions between pesticides and warming is important to improve ecological risk assessment in a warming world. Current insights are almost exclusively based on studies that exposed animals simultaneously to both warming and a pesticide and focused on effects during the pesticide exposure period and within a single developmental stage. We studied two ignored aspects of the interplay between warming and pesticide exposure: (i) the role of delayed effects after the pesticide exposure period, and (ii) the dependence on the developmental stage. We carried out a longitudinal experiment from the egg stage to the adult stage in the mosquito Culex pipiens where we crossed a warming treatment (20 °C vs 24 °C) with 48 h exposures to the pesticide chlorpyrifos in three developmental stages (early L1 larvae, late L4 larvae and adults). Chlorpyrifos induced mild to moderate mortality in all developmental stages (10-30%). A key finding was that warming shaped the chlorpyrifos-induced mortality but in opposite directions between stages. Chlorpyrifos was 7% less toxic under warming in L1 larvae, yet more toxic under warming in L4 larvae (22%) and in adult males (33%), while toxicity did not change under warming in adult females. We hypothesize that the general, stage-specific differences in the effects of warming on body size (increased size in early larvae, decreased size in later stages) caused the reversal of the effects of warming on toxicity between stages. Previous larval exposure to chlorpyrifos caused delayed effects that strongly reduced survival to the adult stage (Ì°25% at 24 °C). Notably, warming also modulated these delayed mortality effects in opposite ways between developmental stages, matching the patterns of mortality during the pesticide exposure periods. Integrating the general stage-specific patterns of how warming shapes body size is important to advance our mechanistic understanding of the interactions between pesticides and warming.

Warming increases chlorpyrifos effects on predator but not anti-predator behaviours.

Recent insights indicate that negative effects of pesticides on aquatic biota occur at concentrations that current legislation considers environmentally protective. We here address two, potentially interacting, mechanisms that may contribute to the underestimation of the impact of sublethal pesticide effects in single species tests at room temperature: the impairment of predator and antipredator behaviours and the stronger impact of organophosphate pesticides at higher temperatures. To address these issues we assessed the effects of chlorpyrifos on the predator and antipredator behaviours of larvae of the damselfly Ischnura elegans, important intermediate predators in aquatic food webs, in a common-garden warming experiment with replicated low- and high-latitude populations along the latitudinal gradient of this species in Europe. Chlorpyrifos reduced the levels of predator behavioural endpoints, and this reduction was stronger at the higher temperature for head orientations and feeding strikes. Chlorpyrifos also impaired two key antipredator behavioural endpoints, activity reductions in response to predator cues were smaller in the presence of chlorpyrifos, and chlorpyrifos caused a lower escape swimming speed; these effects were independent of temperature. This suggests chlorpyrifos may impact food web interactions by changing predator-prey interactions both with higher (predators) and lower trophic levels (food). Given that only the interaction with the lower trophic level was more impaired at higher temperatures, the overall pesticide-induced changes in food web dynamics may be strongly temperature-dependent. These findings were consistent in damselflies from low- and high-latitude populations, illustrating that thermal adaptation will not mitigate the increased toxicity of pesticides at higher temperatures. Our study not only underscores the relevance of including temperature and prey-predator interactions in ecological risk assessment but also their potential interplay and thereby highlights the complexity of contaminant effects on predator-prey interactions being differentially temperature-dependent pending on the trophic level.

Local adaptation and the potential effects of a contaminant on predator avoidance and antipredator responses under global warming: a space-for-time substitution approach.

The ability to deal with temperature-induced changes in interactions with contaminants and predators under global warming is one of the outstanding, applied evolutionary questions. For this, it is crucial to understand how contaminants will affect activity levels, predator avoidance and antipredator responses under global warming and to what extent gradual thermal evolution may mitigate these effects. Using a space-for-time substitution approach, we assessed the potential for gradual thermal evolution shaping activity (mobility and foraging), predator avoidance and antipredator responses when Ischnura elegans damselfly larvae were exposed to zinc in a common-garden warming experiment at the mean summer water temperatures of shallow water bodies at southern and northern latitudes (24 and 20°C, respectively). Zinc reduced mobility and foraging, predator avoidance and escape swimming speed. Importantly, high-latitude populations showed stronger zinc-induced reductions in escape swimming speed at both temperatures, and in activity levels at the high temperature. The latter indicates that local thermal adaptation may strongly change the ecological impact of contaminants under global warming. Our study underscores the critical importance of considering local adaptation along natural gradients when integrating biotic interactions in ecological risk assessment, and the potential of gradual thermal evolution mitigating the effects of warming on the vulnerability to contaminants.

Extreme temperatures in the adult stage shape delayed effects of larval pesticide stress: a comparison between latitudes.

Global warming and pesticide pollution are major threats for aquatic biodiversity. Yet, how pesticide effects are influenced by the increased frequency of extreme temperatures under global warming and how local thermal adaptation may mitigate these effects is unknown. We therefore investigated the combined impact of larval chlorpyrifos exposure, larval food stress and adult heat exposure on a set of fitness-related traits in replicated low- and high-latitude populations of the damselfly Ischnura elegans. Larval pesticide exposure resulted in lighter adults with a higher water content, lower fat content, higher Hsp70 levels and a lower immune function (PO activity). Heat exposure reduced water content, mass, fat content and flying ability. Importantly, both stressors interacted across metamorphosis: adult heat exposure lowered the reduction of fat content, and generated a stronger decrease in PO activity in pesticide-exposed animals. Larval pesticide exposure and larval food stress also reduced the defense response to the adult heat stress in terms of increased Hsp70 levels. In line with strong life history differences in the unstressed control situation, high-latitude animals were less sensitive to food stress (body mass and water content), but more sensitive to pesticide stress (development time and PO activity) and heat exposure (PO activity and Hsp70 levels). While low-latitude adults could better withstand the extreme temperature as suggested by the weaker increase in Hsp70, heat exposure similarly affected the delayed effects of larval pesticide exposure at both latitudes. Our study highlighted two key findings relevant for ecological risk assessment under global warming. Firstly, the delayed effects of larval pesticide exposure on adult damselflies depended upon subsequent adult heat exposure, indicating that larval pesticide stress and adult heat stress interacted across metamorphosis. Secondly, low- and high-latitude animals responded differently to the imposed stressors, highlighting that intraspecific evolution along natural thermal gradients may shape sensitivity to pesticides.

Susceptibility to a metal under global warming is shaped by thermal adaptation along a latitudinal gradient.

Global warming and contamination represent two major threats to biodiversity that have the potential to interact synergistically. There is the potential for gradual local thermal adaptation and dispersal to higher latitudes to mitigate the susceptibility of organisms to contaminants and global warming at high latitudes. Here, we applied a space-for-time substitution approach to study the thermal dependence of the susceptibility of Ischnura elegans damselfly larvae to zinc in a common garden warming experiment (20 and 24 °C) with replicated populations from three latitudes spanning >1500 km in Europe. We observed a striking latitude-specific effect of temperature on the zinc-induced mortality pattern; local thermal adaptation along the latitudinal gradient made Swedish, but not French, damselfly larvae more susceptible to zinc at 24 °C. Latitude- and temperature-specific differences in zinc susceptibility may be related to the amount of energy available to defend against and repair damage since Swedish larvae showed a much stronger zinc-induced reduction of food intake at 24 °C. The pattern of local thermal adaptation indicates that the predicted temperature increase of 4 °C by 2100 will strongly magnify the impact of a contaminant such as zinc at higher latitudes unless there is thermal evolution and/or migration of lower latitude genotypes. Our results underscore the critical importance of studying the susceptibility to contaminants under realistic warming scenarios taking into account local thermal adaptation across natural temperature gradients.