Pharmaceutical Pollution Alters the Structure of Freshwater Communities and Hinders Their Recovery from a Fish Predator.

Freshwater ecosystems are under threat from rising pharmaceutical pollution. While such pollutants are known to elicit biological effects on organisms, we have limited knowledge on how these effects might cascade through food-webs, disrupt ecological processes, and shape freshwater communities. In this study, we used a mesocosm experiment to explore how the community impacts of a top-order predator, the eastern mosquitofish (Gambusia holbrooki), are mediated by exposure to environmentally relevant low (measured concentration: ∼10 ng/L) and high concentrations (∼110 ng/L) of the pervasive pharmaceutical pollutant fluoxetine. We found no evidence that exposure to fluoxetine altered the consumptive effects of mosquitofish on zooplankton. However, once mosquitofish were removed from the mesocosms, zooplankton abundance recovered to a greater extent in control mesocosms compared to both low and high fluoxetine-exposed mesocosms. By the end of the experiment, this resulted in fundamental differences in community structure between the control and fluoxetine-treated mesocosms. Specifically, the control mesocosms were characterized by higher zooplankton abundances and lower algal biomass, whereas mesocosms exposed to either low or high concentrations of fluoxetine had lower zooplankton abundances and higher algal biomass. Our results suggest that fluoxetine, even at very low concentrations, can alter aquatic communities and hinder their recovery from disturbances.

Temperature change exerts sex-specific effects on behavioural variation.

Temperature is a key factor mediating organismal fitness and has important consequences for species' ecology. While the mean effects of temperature on behaviour have been well-documented in ectotherms, how temperature alters behavioural variation among and within individuals, and whether this differs between the sexes, remains unclear. Such effects likely have ecological and evolutionary consequences, given that selection acts at the individual level. We investigated the effect of temperature on individual-level behavioural variation and metabolism in adult male and female Drosophila melanogaster (n = 129), by taking repeated measures of locomotor activity and metabolic rate at both a standard temperature (25°C) and a high temperature (28°C). Males were moderately more responsive in their mean activity levels to temperature change when compared to females. However, this was not true for either standard or active metabolic rate, where no sex differences in thermal metabolic plasticity were found. Furthermore, higher temperatures increased both among- and within-individual variation in male, but not female, locomotor activity. Given that behavioural variation can be critical to population persistence, we suggest that future studies test whether sex differences in the amount of behavioural variation expressed in response to temperature change may result in sex-specific vulnerabilities to a warming climate.

Social context mediates the expression of a personality trait in a gregarious lizard.

The social environment is a key factor that influences behavioural traits across a wide array of species. Yet, when investigating individual differences in behaviour, studies tend to measure animals in isolation from other conspecifics-even in social species. Surprisingly, whether behavioural traits measured in isolation are predictive of individual-level behaviour when in social groups is still poorly understood. Here, we repeatedly measured risk-taking behaviour (i.e. boldness; 741 total trials) in both the presence and absence of conspecifics in a social lizard, the delicate skink (Lampropholis delicata). Further, we manipulated food availability during group trials to test whether the effect of the social environment on risk-taking behaviour was mediated by competition over resources. Using 105 lizards collected from three independent populations, we found that individual risk-taking behaviour was repeatable when measured in either social isolation or within groups both with and without food resources available. However, lizards that were bolder during individual trials were not also bolder when in groups, regardless of resource availability. This was largely driven by individual differences in social behavioural plasticity, whereby individual skinks responded differently to the presence of conspecifics. Together, this resulted in a rank order change of individual behavioural types across the social conditions. Our results highlight the importance of the social environment in mediating animal personality traits across varying levels of resource availability. Further, these findings suggest that behavioural traits when measured in isolation, may not reflect individual variation in behaviour when measured in more ecologically realistic social groups.

Psychoactive pollution suppresses individual differences in fish behaviour.

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.

Context is Key: Social Environment Mediates the Impacts of a Psychoactive Pollutant on Shoaling Behavior in Fish.

Behavior-modifying drugs, such as antidepressants, are increasingly being detected in waterways and aquatic wildlife around the globe. Typically, behavioral effects of these contaminants are assessed using animals tested in social isolation. However, for group-living species, effects seen in isolation may not reflect those occurring in realistic social settings. Furthermore, interactions between chemical pollution and other stressors, such as predation risk, are seldom considered. This is true even though animals in the wild are rarely, if ever, confronted by chemical pollution as a single stressor. Here, in a 2 year multigenerational experiment, we tested for effects of the antidepressant fluoxetine (measured concentrations [±SD]: 42.27 ± 36.14 and 359.06 ± 262.65 ng/L) on shoaling behavior in guppies (Poecilia reticulata) across different social contexts and under varying levels of perceived predation risk. Shoaling propensity and shoal choice (choice of groups with different densities) were assessed in a Y-maze under the presence of a predatory or nonpredatory heterospecific, with guppies tested individually and in male-female pairs. When tested individually, no effect of fluoxetine was seen on shoaling behavior. However, in paired trials, high-fluoxetine-exposed fish exhibited a significantly greater shoaling propensity. Hence, effects of fluoxetine were mediated by social context, highlighting the importance of this fundamental but rarely considered factor when evaluating impacts of environmental pollution.

Reproduction in a polluted world: implications for wildlife.

Environmental pollution is an increasing problem for wildlife globally. Animals are confronted with many different forms of pollution, including chemicals, light, noise, and heat, and these can disrupt critical biological processes such as reproduction. Impacts on reproductive processes can dramatically reduce the number and quality of offspring produced by exposed individuals, and this can have further repercussions on the ecology and evolution of affected populations. Here, we illustrate how environmental pollutants can affect various components of reproduction in wildlife, including direct impacts on reproductive physiology and development, consequences for gamete quality and function, as well as effects on sexual communication, sexual selection, and parental care. We follow with a discussion of the broader ecological and evolutionary consequences of these effects on reproduction and suggest future directions that may enable us to better understand and address the effects of environmental pollution.

Long-Term Pharmaceutical Contamination and Temperature Stress Disrupt Fish Behavior.

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 behaviors 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 behavior. However, both stressors had independent impacts. Fluoxetine exposure resulted in increased male coercive copulatory behavior, while fish activity levels were unaffected. Under cold-temperature stress, both sexes were less active and males exhibited less frequent reproductive behaviors. Our results demonstrate that long-term exposure to a common pharmaceutical pollutant and acute temperature stress alter fundamental fitness-related behaviors in fish, potentially shifting population dynamics in contaminated ecosystems.

Field-realistic antidepressant exposure disrupts group foraging dynamics in mosquitofish.

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.

Antidepressants in Surface Waters: Fluoxetine Influences Mosquitofish Anxiety-Related Behavior at Environmentally Relevant Levels.

Pharmaceutical contamination is an increasing problem globally. In this regard, the selective serotonin reuptake inhibitors (SSRIs)-a group of antidepressants-are particularly concerning. By disrupting the serotonergic system, SSRIs have the potential to affect ecologically important behaviors in exposed wildlife. Despite this, the nature and magnitude of behavioral perturbations resulting from environmentally relevant SSRI exposure among species is poorly understood. Accordingly, we investigated the effects of two field-realistic levels of the SSRI fluoxetine (61 and 352 ng/L) on sociability and anxiety-related behaviors in eastern mosquitofish ( Gambusia holbrooki) for 28 days. Additionally, we measured whole-body tissue concentrations of fluoxetine and norfluoxetine. We found that fluoxetine altered anxiety-related behavior but not sociability. Specifically, female fish showed reduced anxiety-related behavior at the lower treatment level, while males showed an increase at the higher treatment level. In addition, we report a biomass-dependent and sex-specific accumulation of fluoxetine and norfluoxetine, with smaller fish showing higher relative tissue concentrations, with this relationship being more pronounced in males. Our study provides evidence for nonmonotonic and sex-specific effects of fluoxetine exposure at field-realistic concentrations. More broadly, our study demonstrated that neuroactive pharmaceuticals, such as fluoxetine, can affect aquatic life by causing subtle but important shifts in ecologically relevant behaviors.

Impacts of Exposure to Ultraviolet Radiation and an Agricultural Pollutant on Morphology and Behavior of Tadpoles (Limnodynastes tasmaniensis).

Amphibians are the most threatened vertebrate class globally. Multiple factors have been implicated in their global decline, and it has been hypothesized that interactions between stressors may be a major cause. Increased ultraviolet (UV) radiation, as a result of ozone depletion, has been identified as one such stressor. Exposure to UV radiation has been shown to have detrimental effects on amphibians and can exacerbate the effects of other stressors, such as chemical pollutants. Chemical pollution has likewise been recognized as a major factor contributing to amphibian declines, particularly, endocrine-disrupting chemicals. In this regard, 17ß-trenbolone is a potent anabolic steroid used in the agricultural industry to increase muscle mass in cattle and has been repeatedly detected in the environment where amphibians live and breed. At high concentrations, 17ß-trenbolone has been shown to impact amphibian survival and gonadal development. In the present study, we investigated the effects of environmentally realistic UV radiation and 17ß-trenbolone exposure, both in isolation and in combination, on the morphology and behavior of tadpoles (Limnodynastes tasmaniensis). We found that neither stressor in isolation affected tadpoles, nor did we find any interactive effects. The results from our 17ß-trenbolone treatment are consistent with recent research suggesting that, at environmentally realistic concentrations, tadpoles may be less vulnerable to this pollutant compared to other vertebrate classes. The absence of UV radiation-induced effects found in the present study could be due to species-specific variation in susceptibility, as well as the dosage utilized. We suggest that future research should incorporate long-term studies with multiple stressors to accurately identify the threats to, and subsequent consequences for, amphibians under natural conditions. Environ Toxicol Chem 2024;43:1615-1626. © 2024 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Widespread psychoactive pollutant augments daytime restfulness and disrupts diurnal activity rhythms in fish.

Pharmaceutical pollution is a major driver of global change, with the capacity to alter key behavioural and physiological traits in exposed animals. Antidepressants are among the most commonly detected pharmaceuticals in the environment. Despite well-documented pharmacological effects of antidepressants on sleep in humans and other vertebrates, very little is known about their ecologically relevant impacts as pollutants on non-target wildlife. Accordingly, we investigated the effects of acute 3-day exposure of eastern mosquitofish (Gambusia holbrooki) to field-realistic levels (nominal concentrations: 30 and 300 ng/L) of the widespread psychoactive pollutant, fluoxetine, on diurnal activity patterns and restfulness, as indicators of disruptions to sleep. We show that exposure to fluoxetine disrupted diel activity patterns, which was driven by augmentation of daytime inactivity. Specifically, unexposed control fish were markedly diurnal, swimming farther during the day and exhibiting longer periods and more bouts of inactivity at night. However, in fluoxetine-exposed fish, this natural diel rhythm was eroded, with no differences in activity or restfulness observed between the day and night. As a misalignment in the circadian rhythm has been shown to adversely affect fecundity and lifespan in animals, our findings reveal a potentially serious threat to the survival and reproductive success of pollutant-exposed wildlife.

Sex differences in the predictability of risk-taking behavior.

Recent research has found that individuals often vary in how consistently they express their behavior over time (i.e., behavioral predictability) and suggested that these individual differences may be heritable. However, little is known about the intrinsic factors that drive variation in the predictability of behavior. Indeed, whether variation in behavioral predictability is sex-specific is not clear. This is important, as behavioral predictability has been associated with vulnerability to predation, suggesting that the predictability of behavioral traits may have key fitness implications. We investigated whether male and female eastern mosquitofish (Gambusia holbrooki) differed in the predictability of their risk-taking behavior. Specifically, over a total of 954 behavioral trials, we repeatedly measured risk-taking behavior with three commonly used assays-refuge-use, thigmotaxis, and foraging latency. We predicted that there would be consistent sex differences in both mean-level risk-taking behavior and behavioral predictability across the assays. We found that risk-taking behavior was repeatable within each assay, and that some individuals were consistently bolder than others across all three assays. There were also consistent sex differences in mean-level risk-taking behavior, with males being bolder across all three assays compared to females. In contrast, both the magnitude and direction of sex differences in behavioral predictability were assay-specific. Taken together, these results highlight that behavioral predictability may be independent from underlying mean-level behavioral traits and suggest that males and females may differentially adjust the consistency of their risk-taking behavior in response to subtle changes in environmental conditions.

Bigger and bolder: Widespread agricultural pollutant 17ß-trenbolone increases growth and alters behaviour in tadpoles (Litoria ewingii).

Endocrine-disrupting chemicals-compounds that directly interfere with the endocrine system of exposed animals-are insidious environmental pollutants that can disrupt hormone function, even at very low concentrations. The dramatic impacts that some endocrine-disrupting chemicals can have on the reproductive development of wildlife are well documented. However, the potential of endocrine-disrupting chemicals to disrupt animal behaviour has received far less attention, despite the important links between behavioural processes and population-level fitness. Accordingly, we investigated the impacts of 14 and 21-day exposure to two environmentally realistic levels of 17ß-trenbolone (4.6 and 11.2 ng/L), a potent endocrine-disrupting steroid and agricultural pollutant, on growth and behaviour in tadpoles of an anuran amphibian, the southern brown tree frog (Litoria ewingii). We found that 17ß-trenbolone altered morphology, baseline activity and responses to a predatory threat, but did not affect anxiety-like behaviours in a scototaxis assay. Specifically, we found that tadpoles exposed to our high-17ß-trenbolone treatment were significantly longer and heavier at 14 and 21 days. We also found that tadpoles exposed to 17ß-trenbolone showed higher levels of baseline activity, and significantly reduced their activity following a simulated predator strike. These results provide insights into the wider repercussions of agricultural pollutants on key developmental and behavioural traits in aquatic species, and demonstrate the importance of behavioural studies in the ecotoxicological field.

Comparison of triploid and diploid rainbow trout (Oncorhynchus mykiss) fine-scale movement, migration and catchability in lowland lakes of western Washington.

Fisheries managers stock triploid (i.e., infertile, artificially produced) rainbow trout Oncorhynchus mykiss in North American lakes to support sport fisheries while minimizing the risk of genetic introgression between hatchery and wild trout. In Washington State, the Washington Department of Fish and Wildlife (WDFW) allocates approximately US $3 million annually to stock hatchery-origin rainbow trout in > 600 lakes, yet only about 10% of them are triploids. Many lakes in Washington State drain into waters that support wild anadromous steelhead O. mykiss that are listed as threatened under the U.S. Endangered Species Act. As a result, there is a strong interest in understanding the costs and benefits associated with stocking sterile, triploid rainbow trout as an alternative to traditional diploids. The objectives of this study were to compare triploid and diploid rainbow trout in terms of: (1) contribution to the sport fishery catch, (2) fine-scale movements within the study lakes, (3) rate of emigration from the lake, and (4) natural mortality. Our results demonstrated that triploid and diploid trout had similar day-night distribution patterns, but triploid trout exhibited a lower emigration rate from the lake and lower catch rates in some lakes. Overall, triploid rainbow trout represent a viable alternative to stocking of diploids, especially in lakes draining to rivers, because they are sterile, have comparable home ranges, and less often migrate.

Sex-specific effects of psychoactive pollution on behavioral individuality and plasticity in fish.

The global rise of pharmaceutical contaminants in the aquatic environment poses a serious threat to ecological and evolutionary processes. Studies have traditionally focused on the collateral (average) effects of psychoactive pollutants on ecologically relevant behaviors of wildlife, often neglecting effects among and within individuals, and whether they differ between males and females. We tested whether psychoactive pollutants have sex-specific effects on behavioral individuality and plasticity in guppies (Poecilia reticulata), a freshwater species that inhabits contaminated waterways in the wild. Fish were exposed to fluoxetine (Prozac) for 2 years across multiple generations before their activity and stress-related behavior were repeatedly assayed. Using a Bayesian statistical approach that partitions the effects among and within individuals, we found that males-but not females-in fluoxetine-exposed populations differed less from each other in their behavior (lower behavioral individuality) than unexposed males. In sharp contrast, effects on behavioral plasticity were observed in females-but not in males-whereby exposure to even low levels of fluoxetine resulted in a substantial decrease (activity) and increase (freezing behavior) in the behavioral plasticity of females. Our evidence reveals that psychoactive pollution has sex-specific effects on the individual behavior of fish, suggesting that males and females might not be equally vulnerable to global pollutants.

Micropollutants.

In this Quick guide, Bertram et al. highlight the growing problem of micropollutants - a wide array of natural and synthetic organic compounds found in the environment.

Multi-generational impacts of exposure to antidepressant fluoxetine on behaviour, reproduction, and morphology of freshwater snail Physa acuta.

Contamination of the environment by pharmaceutical pollutants poses an increasingly critical threat to aquatic ecosystems around the world. This is particularly true of psychoactive compounds, such as antidepressant drugs, which have become ubiquitous contaminants and have been demonstrated to modify aquatic animal behaviours at very low concentrations (i.e. ng/L). Despite raising risks to the hydrosphere, there is a notable paucity of data on the long term, multigenerational effects of antidepressants at environmentally realistic concentrations. Moreover, current research has predominantly focused on mean-level effects, with little research on variation among and within individuals when considering key behavioural traits. In this work, we used a multigenerational exposure of a freshwater snail (Physa acuta) to an environmentally relevant concentration of the antidepressant fluoxetine (mean measured concentration: 32.7 ng/L, SE: 2.3). The snails were allowed to breed freely in large mesocosm populations over 3 years. Upon completion of the exposure, we repeatedly measured the locomotory activity (624 measures total), reproductive output (234 measures total) as well as morphometric endpoints (78 measures total). While we found no mean-level differences between treatments in locomotory activities, we did find that fluoxetine exposed snails (n = 46) had significantly reduced behavioural plasticity (i.e. VW; within-individual variation) in activity levels compared to unexposed snails (n = 32). As a result, fluoxetine exposed snails demonstrated significant behavioural repeatability, which was not the case for unexposed snails. Further, we report a reduction in egg mass production in fluoxetine exposed snails, and a marginally non-significant difference in morphology between treatment groups. These results highlight the potential detrimental effects of long-term fluoxetine exposure on non-target organisms at environmentally realistic dosages. Additionally, our findings demonstrate the underappreciated potential for psychoactive contaminants to have impacts beyond mean-level effects, with consequences for population resilience to current and future environmental challenges.

Effects of the agricultural pollutant 17ß-trenbolone on morphology and behaviour of tadpoles (Limnodynastes tasmaniensis).

Pollutants, such as endocrine disrupting chemicals (EDCs), are increasingly being detected in organisms and ecosystems globally. Agricultural activities, including the use of hormonal growth promotants (HGPs), are a major source of EDC contamination. One potent EDC that enters into the environment through the use of HGPs is 17ß-trenbolone. Despite EDCs being repeatedly shown to affect reproduction and development, comparatively little is known regarding their effects on behaviour. Amphibians, one of the most imperilled vertebrate taxa globally, are at particular risk of exposure to such pollutants as they often live and breed near agricultural operations. Yet, no previous research on amphibians has explored the effects of 17ß-trenbolone exposure on foraging or antipredator behaviour, both of which are key fitness-related behavioural traits. Accordingly, we investigated the impacts of 28-day exposure to two environmentally realistic concentrations of 17ß-trenbolone (average measured concentrations: 10 and 66 ng/L) on the behaviour and growth of spotted marsh frog tadpoles (Limnodynastes tasmaniensis). Contrary to our predictions, there was no significant effect of 17ß-trenbolone exposure on tadpole growth, antipredator response, anxiety-like behaviour, or foraging. We hypothesise that the differences in effects found between this study and those conducted on fish may be due to taxonomic differences and/or the life stage of the animals used, and suggest further research is needed to investigate the potential for delayed manifestation of the effects of 17ß-trenbolone exposure.

Predicting the impacts of chemical pollutants on animal groups.

Chemical pollution is among the fastest-growing agents of global change. Synthetic chemicals with diverse modes-of-action are being detected in the tissues of wildlife and pervade entire food webs. Although such pollutants can elicit a range of sublethal effects on individual organisms, research on how chemical pollutants affect animal groups is severely lacking. Here we synthesise research from two related, but largely segregated fields - ecotoxicology and behavioural ecology - to examine pathways by which chemical contaminants could disrupt processes that govern the emergence, self-organisation, and collective function of animal groups. Our review provides a roadmap for prioritising the study of chemical pollutants within the context of sociality and highlights important methodological advancements for future research.