Animal behaviour is remarkably sensitive to disruption by chemical pollution, with widespread implications for ecological and evolutionary processes in contaminated wildlife populations. However, conventional approaches applied to study the impacts of chemical pollutants on wildlife behaviour seldom address the complexity of natural environments in which contamination occurs. The aim of this review is to guide the rapidly developing field of behavioural ecotoxicology towards increased environmental realism, ecological complexity, and mechanistic understanding. We identify research areas in ecology that to date have been largely overlooked within behavioural ecotoxicology but which promise to yield valuable insights, including within-and among-individual variation, social networks and collective behaviour, and multi-stressor interactions. Further, we feature methodological and technological innovations that enable the collection of data on pollutant-induced behavioural changes at an unprecedented resolution and scale in the laboratory and the field. In an era of rapid environmental change, there is an urgent need to advance our understanding of the real-world impacts of chemical pollution on wildlife behaviour. This review therefore provides a roadmap of the major outstanding questions in behavioural ecotoxicology and highlights the need for increased cross-talk with other disciplines in order to find the answers.

Environmental contamination by pharmaceuticals is global, substantially altering crucial behaviours in animals and impacting on their reproduction and survival. A key question is whether the consequences of these pollutants extend beyond mean behavioural changes, restraining differences in behaviour between individuals. In a controlled, two-year, multigenerational experiment with independent mesocosm populations, we exposed guppies ( Poecilia reticulata ) to environmentally realistic levels of the ubiquitous pollutant fluoxetine (Prozac). Fish (unexposed: n = 59, low fluoxetine: n = 57, high fluoxetine: n = 58) were repeatedly assayed on four separate occasions for activity and risk-taking behaviour. Fluoxetine homogenized individuals' activity, with individual variation in populations exposed to even low concentrations falling to less than half that in unexposed populations. To understand the proximate mechanism underlying these changes, we tested the relative contribution of variation within and between individuals to the overall decline in individual variation. We found strong evidence that fluoxetine erodes variation in activity between but not within individuals, revealing the hidden consequences of a ubiquitous contaminant on phenotypic variation in fish—likely to impair adaptive potential to environmental change.

Psychoactive pollutants, such as antidepressants, are increasingly detected in the environment. Mounting evidence suggests that such pollutants can disrupt the behaviour of non-target species. Despite this, few studies have considered how the response of exposed organisms might be mediated by social context. To redress this, we investigated the impacts of two environmentally realistic concentrations of a pervasive antidepressant pollutant, fluoxetine, on foraging behaviour in fish ( Gambusia holbrooki ), tested individually or in a group. Fluoxetine did not alter behaviour of solitary fish. However, in a group setting, fluoxetine exposure disrupted the frequency of aggressive interactions and food consumption, with observed effects being contingent on both the mean weight of group members and the level of within-group variation in weight. Our results suggest that behavioural tests in social isolation may not accurately predict the environmental risk of chemical pollutants for group-living species and highlight the potential for social context to mediate the effects of psychoactive pollutants in exposed wildlife.

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Natural environments are subject to a range of anthropogenic stressors, with pharmaceutical pollution being among the fastest-growing agents of global change.However, despite wild animals living in complex multi-stressor environments, interactions between pharmaceutical exposure and other stressors remain poorly understood. Accordingly, we investigated effects of long-term exposure to the pervasive pharmaceutical contaminant fluoxetine (Prozac ® ), and acute temperature stress, on reproductive behaviours and activity levels in the guppy (Poecilia reticulata). Fish were exposed to environmentally realistic fluoxetine concentrations (measured average: 38 or 312 ng/L) or a solvent control for 15 months using a mesocosm system. Additionally, fish were subjected to one of three acute (24 h) temperature treatments: cold stress ( 18°C), heat stress (32 °C) or a control (24 °C). We found no evidence for interactive effects of fluoxetine exposure and temperature stress on guppy behaviour. However, both stressors had independent impacts. Fluoxetine exposure resulted in increased male coercive copulatory behaviour, while fish activity levels were unaffected. Under cold-temperature stress, both sexes were less active and males exhibited less frequent reproductive behaviours. Our results demonstrate that long-term exposure to a common pharmaceutical pollutant, and acute temperature stress, alter fundamental fitnessrelated behaviours in fish, potentially shifting population dynamics in contaminated ecosystems.

Background Globally, there is growing concern over the impacts of pharmaceuticals and drug manufacturing on aquatic animals, and pharmaceuticals are now recognized as contaminants of emerging environmental concern. In recent years, scientists, environmental managers, and policymakers have been interested in using behavioural endpoints for chemical regulation, given their importance for fitness and survival. The body of research on whether and how pharmaceutical exposure alters the behaviour of aquatic animals has grown exponentially, making it difficult to get an overview of the results. With an international spotlight on the management of these environmental threats, synthesizing the currently available data is vital to inform managers and policymakers, as well as highlighting areas where more research is needed. This is a protocol for a systematic evidence map (SEM) and serves as an a priori record of our objectives and methodological decisions. Our objectives are to identify, catalogue, and present primary research articles on the effects of human and veterinary pharmaceuticals on aquatic animal behaviour. Methods The literature search will be conducted using two electronic databases: Web of Science and Scopus, and we will supplement these searches with additional sources. The search string has been developed using a Population–Exposure–Comparison–Outcome (PECO) framework, to capture articles that used an aquatic organism (P, population) to test the effects of a pharmaceutical (E, exposure) on behaviour (O, outcome). Eligible articles must also have a control group (C, comparison). Articles will be screened in two stages, title and abstract, followed by full-text screening before data extraction. Decision trees have been designed a priori to appraise articles for eligibility at both stages of screening. At both stages, screening each article will be completed by two independent reviewers. Study validity will be appraised but not used as a basis for article inclusion. The information extracted from the eligible articles, along with bibliometric data, will be mapped and displayed. All data associated with this SEM will be publicly available through the Open Science Framework (OSF) and a future project webpage.

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