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Pharmaceutical pollution poses an increasing threat to global wildlife populations. Psychoactive pharmaceutical pollutants (e.g. antidepressants, anxiolytics) are a distinctive concern owing to their ability to act on neural pathways that mediate fitness-related behavioural traits. However, despite increasing research efforts, very little is known about how these drugs might influence the behaviour and survival of species in the wild. Here, we capitalize on the development of novel slow-release pharmaceutical implants and acoustic telemetry tracking tools to reveal that exposure to environmentally relevant concentrations of the benzodiazepine pollutant temazepam alters movement dynamics and decreases the migration success of brown trout ( Salmo trutta ) smolts in a natural lake system. This effect was potentially owing to temazepam-exposed fish suffering increased predation compared with unexposed conspecifics, particularly at the river-lake confluence. These findings underscore the ability of pharmaceutical pollution to alter key fitness-related behavioural traits under natural conditions, with likely negative impacts on the health and persistence of wildlife populations.

Artificial light at night (ALAN) can disrupt the natural behaviour, physiology, and circadian rhythms of organisms exposed to it, and therefore presents a significant and widespread ecological concern. ALAN typically comprises a wide range of wavelengths, and different wavelengths have different effects on circadian clocks. In the animals investigated thus far, short and middle wavelengths are intensely involved in synchronisation and entrainment, but we still have a poor understanding of how different wavelengths might affect behaviour when animals are exposed to ALAN, in particular whether some wavelengths are disproportionally detrimental. This experiment examined the direct and transgenerational effects of 10 different wavelength treatments of ALAN on behaviour in zebrafish (Danio rerio), a diurnally active model organism. Across a 10-day period, female zebrafish were exposed to either a monochromatic wavelength, white light ALAN, or to a control treatment, and the individual impacts of each treatment on locomotion and anxiety-like behaviours were examined both for solitary fish and fish in groups. We found the strongest impact at short wavelengths (365 to 470 nm), with individuals and groups of zebrafish showing more anxiety-like behaviour after fewer nights of ALAN exposure relative to the other wavelengths. Furthermore, F1 offspring born from ALAN-exposed mothers displayed less frequent movement and shorter movement distances despite never being exposed to ALAN themselves, regardless of the spectral treatment. Our results highlight both the specific and broad-spectrum potential for ALAN to cause disruption to locomotion in adult zebrafish and their offspring., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

In recent years, the sediment compartment has gained more attention when performing toxicity tests, with a growing emphasis on gaining more ecological relevance in testing. Though many standard guidelines recommend using artificially formulated sediment, most sediment studies are using natural sediment collected in the field. Although the use of natural field-collected sediment contributes to more environmentally realistic exposure scenarios and higher well-being for sediment-dwelling organisms, it lowers comparability and reproducibility among studies as a result of, for example, differences in the base sediment depending on sampling site, background contamination, particle size distribution, or organic matter content. The aim of this methodology contribution is to present and discuss best practices related to collecting, handling, describing, and applying natural field-collected sediment in ecotoxicological testing. We propose six recommendations: (1) natural sediment should be collected at a well-studied site, historically and by laboratory analysis; (2) larger quantities of sediment should be collected and stored prior to initiation of an experiment to ensure a uniform sediment base; (3) any sediment used in ecotoxicological testing should be characterized, at the very least, for its water content, organic matter content, pH, and particle size distribution; (4) select spiking method, equilibration time, and experimental setup based on the properties of the contaminant and the research question; (5) include control-, treated similarly to the spiked sediment, and solvent control sediment when appropriate; and (6) quantify experimental exposure concentrations in the overlying water, porewater (if applicable), and bulk sediment at least at the beginning and the end of each experiment. Environ Toxicol Chem 2024;43:1757-1766. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC., (© 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.)

Pharmaceuticals and personal care products (PPCPs) occur as variable mixtures in surface waters receiving discharges of human and animal wastes. A key question identified a decade ago is how to assess the effects of long-term exposures of these PPCP mixtures on nontarget organisms. We review the recent progress made on assessing the aquatic ecotoxicity of PPCP mixtures-with a focus on active pharmaceutical ingredients-and the challenges and research needs that remain. New knowledge has arisen from the use of whole-mixture testing combined with component-based approaches, and these studies show that mixtures often result in responses that meet the concentration addition model. However, such studies have mainly been done on individual species over shorter time periods, and longer-term, multispecies assessments remain limited. The recent use of targeted and nontargeted gene analyses has improved our understanding of the diverse pathways that are impacted, and there are promising new "read-across" methods that use mammalian data to predict toxicity in wildlife. Risk assessments remain challenging given the paucity of ecotoxicological and exposure data on PPCP mixtures. As such, the assessment of PPCP mixtures in aquatic environments should remain a priority given the potential for additive-as well as nontarget-effects in nontarget organisms. In addition, we need to improve our understanding of which species, life stages, and relevant endpoints are most sensitive to which types of PPCP mixtures and to expand our knowledge of environmental PPCP levels in regions of the globe that have been poorly studied to date. We recommend an increased use of new approach methodologies, in particular "omics," to advance our understanding of the molecular mechanics of mixture effects. Finally, we call for systematic research on the role of PPCP mixtures in the development of antimicrobial resistance. Environ Toxicol Chem 2024;43:549-558. © 2023 SETAC., (© 2023 SETAC.)

Animal movement is a multifaceted process that occurs for multiple reasons with powerful consequences for food web and ecosystem dynamics. New paradigms and technical innovations have recently pervaded the field, providing increasingly powerful means to deliver fine-scale movement data, attracting renewed interest. Specifically in the aquatic environment, tracking with acoustic telemetry now provides integral spatiotemporal information to follow individual movements in the wild. Yet, this technology also holds great promise for experimental studies, enhancing our ability to truly establish cause-and-effect relationships. Here, we argue that ponds with well-defined borders (i.e. "islands in a sea of land") are particularly well suited for this purpose. To support our argument, we also discuss recent experiences from studies conducted in an innovative experimental infrastructure, composed of replicated ponds equipped with modern aquatic telemetry systems that allow for unparalleled insights into the movement patterns of individual animals., (© 2023. BioMed Central Ltd., part of Springer Nature.)

Internal, slow-release implants can be an effective way to manipulate animal physiology or deliver a chemical exposure over long periods of time without the need for an exogenous exposure route. Slow-release implants involve dissolving a compound in a lipid-based carrier, which is inserted into the body of an organism. However, the release kinetics of the compound from the implant to body tissues also requires careful validation. We tested and validated a slow-release implant methodology for exposing fish to a pharmaceutical pollutant, fluoxetine. We tested two lipid-based carriers (coconut oil or vegetable shortening) in the common roach (Rutilus rutilus). The implants contained either a high (50 μg/g), low (25 μg/g), or control (0 μg/g) concentration of fluoxetine, and we measured tissue uptake in the brain, muscle, and plasma of implanted fish over 25 days. The two carriers released fluoxetine differently over time: coconut oil released fluoxetine in an accelerating manner (tissue uptake displayed a positive quadratic curvature), whereas vegetable shortening released fluoxetine in a decelerating manner (a negative quadratic curvature). For both carrier types, fluoxetine was measured at the highest concentration in the brain, followed by muscle and plasma. By comparing the implant exposures with waterborne exposures in the published literature, we showed that the implants delivered an internal exposure that would be similar if fish were exposed in surface waters containing effluents. Overall, we showed that slow-release internal implants are an effective method for delivering chronic exposures of fluoxetine over at least 1-month time scales. Internal exposures can be an especially powerful experimental tool when coupled with field-based study designs to assess the impacts of pharmaceutical pollutants in complex natural environments. Environ Toxicol Chem 2023;42:1326-1336. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC., (© 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.)

Pharmaceutical pollution represents a rapidly growing threat to ecosystems worldwide. Drugs are now commonly detected in the tissues of wildlife and have the potential to alter the natural expression of behavior, though relatively little is known about how pharmaceuticals impact predator-prey interactions. We conducted parallel laboratory experiments using larval odonates (dragonfly and damselfly nymphs) to investigate the effects of exposure to two pharmaceuticals, cetirizine and citalopram, and their mixture on the outcomes of predator-prey interactions. We found that exposure to both compounds elevated dragonfly activity and impacted their predation success and efficiency in complex ways. While exposure to citalopram reduced predation efficiency, exposure to cetirizine showed varied effects, with predation success being enhanced in some contexts but impaired in others. Our findings underscore the importance of evaluating pharmaceutical effects under multiple contexts and indicate that these compounds can affect predator-prey outcomes at sublethal concentrations., Competing Interests: The authors declare no competing interests., (© 2022 The Author(s).)

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., Competing Interests: Declaration of interests The authors declare no conflicts of interest., (Copyright © 2022 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

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., (© 2022 The Authors. Biological Reviews published by John Wiley & Sons Ltd on behalf of Cambridge Philosophical Society.)

We studied the ecological consequences of widespread caffeine contamination by conducting an experiment focused on changes in the behavioral traits of wild perch (Perca fluviatilis) after waterborne exposure to 10 μg L -1 of caffeine. We monitored fish swimming performance during both light and dark conditions to study the effect of caffeine on fish activity and circadian rhythm, using a novel three-dimensional tracking system that enabled positioning even in complete darkness. All individuals underwent three behavioral trials-before exposure, after 24 h of exposure, and after 5 days of exposure. We did not observe any effect of the given caffeine concentration on fish activity under light or dark conditions. Regardless of caffeine exposure, fish swimming performance was significantly affected by both the light-dark conditions and repeating of behavioral trials. Individuals in both treatments swam significantly more during the light condition and their activity increased with time as follows: before exposure < after 24 h of exposure < after 5 days of exposure. We confirmed that the three-dimensional automated tracking system based on infrared sensors was highly effective for conducting behavioral experiments under completely dark conditions., (© 2022. The Author(s).)

The social environment (i.e., the suite of social interactions that occur among individuals that can result in variation in social ranks) is a commonly overlooked aspect of biology when scientists evaluate the effects of chemical contaminants. The social environment, however, represents the arena in which individual-level performance shapes group- or population-level outcomes and may therefore mediate many of the ultimate consequences of chemicals for wildlife. Here, we evaluated the role that the social environment plays in determining the consequences of pollutant exposure. We exposed groups of juvenile brown trout (Salmo trutta) to an emerging pharmaceutical pollutant that is commonly detected in freshwaters (the benzodiazepine, oxazepam) and allowed them to form dominance hierarchies. Exposure affected dominant and subordinate fish differently, causing fish to become less aggressive at high doses and subordinate fish to become more competitively successful at low doses. These perturbations had further consequences for growth, fin damage, and survival. Exposure also modulated physiological stress in the hierarchy, and social status itself affected how much oxazepam was absorbed in tissues, potentially creating a dynamic feedback loop that further influences the asymmetric effects of exposure on differing social statuses. Many effects followed a "U-shaped" dose-response curve, highlighting the importance of nonlinear, low-dose effects. Altogether, we show that social structure in animal groups can interact with and modulate the effects of an environmental contaminant. We underscore the need to account for an organism's natural ecological context, including their social environment, in future experiments and environmental risk assessments to predict the effects of chemical contaminants on wildlife., (© 2021 Her Majesty the Queen in Right of Canada. Ecological Applications published by Wiley Periodicals LLC on behalf of The Ecological Society of America. Reproduced with the permission of the Minister of Fisheries and Oceans Canada.)

Pharmaceutically active compounds (PhACs) have been shown to accumulate in aquatic and riparian food-webs. Yet, our understanding of how temperature, a key environmental factor in nature, affects uptake, biotransformation, and the subsequent accumulation of PhACs in aquatic organisms is limited. In this study, we tested to what extent bioconcentration of an anxiolytic drugs (temazepam and oxazepam) is affected by two temperature regimes (10 and 20 °C) and how the temperature affects the temazepam biotransformation and subsequent accumulation of its metabolite (oxazepam) in aquatic organisms. We used European perch (Perca fluviatilis) and dragonfly larvae (Sympetrum sp.), which represent predator and prey species of high ecological relevance in food chains of boreal and temperate aquatic ecosystems. Experimental organisms were exposed to target pharmaceuticals at a range of concentrations (0.2-6 µg L -1 ) to study concentration dependent differences in bioconcentration and biotransformation. We found that the bioconcentration of temazepam in perch was significantly reduced at higher temperatures. Also, temperature had a strong effect on temazepam biotransformation in the fish, with the production and subsequent accumulation of its metabolite (oxazepam) being two-fold higher at 20 °C compared to 10 °C. In contrast, we found no temperature dependency for temazepam bioconcentration in dragonfly larvae and no detectable biotransformation of the parent compound that would result in measurable concentrations of oxazepam in this organism. Our results highlight that while organisms may share the same aquatic ecosystem, their exposure to PhACs may change differently across temperature gradients in the environment., (Copyright © 2021 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Municipal wastewater treatment plant (WWTP) effluents are a ubiquitous source of contamination whose impacts on fish and other aquatic organisms span across multiple levels of biological organization. Despite this, few studies have addressed the impacts of WWTP effluents on fish communities, especially during the winter-a season seldom studied. Here, we assessed the impacts of wastewater on fish community compositions and various water quality parameters during the summer and winter along two effluent gradients in Hamilton Harbour, an International Joint Commission Area of Concern in Hamilton, Canada. We found that fish abundance, species richness, and species diversity were generally highest in sites closest to the WWTP outfalls, but only significantly so in the winter. Fish community compositions differed greatly along the effluent gradients, with sites closest and farthest from the outfalls being the most dissimilar. Furthermore, the concentrations of numerous contaminants of emerging concern (CECs) in the final treated effluent were highest during the winter. Water quality of sites closer to the outfalls was poorer than at sites farther away, especially during the winter. We also demonstrated that WWTPs can significantly alter the thermal profile of effluent-receiving environments, increasing temperature by as much as ~9 °C during the winter. Our results suggest that wastewater plumes may act as ecological traps in winter, whereby fish are attracted to the favourable temperatures near WWTPs and are thus exposed to higher concentrations of CECs. This study highlights the importance of winter research as a key predictor in further understanding the impacts of wastewater contamination in aquatic ecosystems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Interest in behavioural ecotoxicology is growing, partly due to technological and computational advances in recording behaviours but also because of improvements of detection capacity facilitating reporting effects at environmentally relevant concentrations. The peer-reviewed literature now contains studies investigating the effects of chemicals, including pesticides and pharmaceuticals, on migration, dispersal, aggression, sociability, reproduction, feeding and anti-predator behaviours in vertebrates and invertebrates. To understand how behavioural studies could be used in regulatory decision-making we: (1) assessed the legal obstacles to using behavioural endpoints in EU chemicals regulation; (2) analysed the known cases of use of behavioural endpoints in EU chemicals regulation; and (3) provided examples of behavioural endpoints of relevance for population level effects. We conclude that the only legal obstacle to the use of behavioural endpoints in EU chemicals regulation is whether an endpoint is considered to be relevant at the population level or not. We also conclude that ecotoxicity studies investigating behavioural endpoints are occasionally used in the EU chemicals regulation, and underscore that behavioural endpoints can be relevant at the population level. To improve the current use of behavioural studies in regulatory decision-making contribution from all relevant stakeholders is required. We have the following recommendations: (1) researchers should conduct robust, well-designed and transparent studies that emphasize the relevance of the study for regulation of chemicals; (2) editors and scientific journals should promote detailed, reliable and clearly reported studies; (3) regulatory agencies and the chemical industry need to embrace new behavioural endpoints of relevance at the population level.

Effluent from wastewater treatment plants (WWTP) contains a complex mixture of contaminants and is a major worldwide source of aquatic pollution. We examined the effects of exposure to treated effluent from a municipal WWTP on the metabolic physiology of bluegill sunfish (Lepomis macrochirus). We studied fish that were wild-caught or experimentally caged (28 d) downstream of the WWTP, and compared them to fish that were caught or caged at clean reference sites. Survival was reduced in fish caged at the effluent-contaminated site compared to those caged at the reference site. Resting rates of O 2 consumption (MO 2 ) were higher in fish from the contaminated site, reflecting a metabolic cost of wastewater exposure. The increases in routine MO 2 did not reduce aerobic scope (difference or quotient of maximal MO 2 and resting MO 2 ), suggesting that physiological compensations accompanied the metabolic costs of wastewater exposure. Fish exposed to wastewater also had larger hearts and livers. The activity of mitochondrial enzymes (cytochrome c oxidase, citrate synthase) per liver mass was unaltered across treatments, so the increased mass of this organ increased its cumulative oxidative capacity in the fish. Wastewater exposure also reduced glycogen content per liver mass. The effects of caging itself, based on comparisons between fish that were wild-caught or caged at clean sites, were generally subtle and not statistically significant. We conclude that exposure to wastewater effluent invokes a metabolic cost that leads to compensatory physiological adjustments that partially offset the detrimental metabolic impacts of exposure., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Although effluent from municipal wastewater treatment plants (WWTPs) is a major stressor in receiving environments, relatively few studies have addressed how its discharge affects natural fish communities. Here, we assessed fish community composition over three years along a gradient of effluent exposure from two distinct WWTPs within an International Joint Commission Area of Concern on the Great Lakes (Hamilton Harbour, Canada). We found that fish communities changed with distance from both WWTPs, and were highly dissimilar between sites that were closest to and furthest from the wastewater outfall. Despite differences in the size and treatment technology of the WWTPs and receiving habitats downstream, we found that the sites nearest the outfalls had the highest fish abundances and contained a common set of signature fish species (i.e., round goby Neogobius melanostomus, green sunfish Lepomis cyanellus). Non-native and stress tolerant species were also more abundant near one of the studied WWTPs when compared to the reference site, and the number of young-of-the-year fish collected did not vary along the effluent exposure gradients. Overall, we show that fish are attracted to wastewater outfalls raising the possibility that these sites may act as an ecological trap., (Crown Copyright © 2019. Published by Elsevier Ltd. All rights reserved.)

Field-based ecotoxicology studies are invaluable for uncovering the effects of contaminants of emerging concern (CECs) on aquatic organisms. However, large-scale exposures are still very rare due to prohibitive costs, the availability of replicated habitats, and the potential for exposure to cause lasting damage to the environment. Here, we evaluated the viability of internal slow-release implants as an alternative method for manipulating CEC exposures in aquatic wildlife using two fat-based carriers (coconut oil and vegetable shortening). We treated roach ( Rutilus rutilus ) with implants containing a high (50 μg/g), low (25 μg/g), or control (0 μg/g) concentration of the behavior-modifying pharmaceutical oxazepam. We then measured oxazepam uptake in four tissues (plasma, muscle, liver, and the brain) over 1 month. The two carriers released oxazepam differently: coconut oil was the superior implant type because it delivered a more consistent dose across time, while vegetable shortening released oxazepam rapidly at the start of the exposure period. For both carriers and treatments, the brain and liver contained the most oxazepam. Overall, the method is a promising technique for controlled manipulations of pharmaceuticals in fish, and we have provided some of the first data on the suitability and contaminant release kinetics from different implant types.

The presence of human pharmaceuticals in the environment has garnered significant research attention because these compounds may exert therapeutic effects on exposed wildlife. Yet, for many compounds, there is still little research documenting their stability in the water column and uptake in organism tissues. Here, we measured the uptake and stability of methylphenidate (Ritalin®, a frequently prescribed central nervous system stimulant) and its primary metabolite, ritalinic acid, in (1) water only or (2) with nine-spine stickleback and water louse. Methylphenidate degraded to ritalinic acid in both studies faster at a higher temperature (20 °C versus 10 °C), with concentrations of ritalinic acid surpassing methylphenidate after 48-100 h, depending on temperature. The concentration of methylphenidate in stickleback was highest at the first sampling point (60 min), while the concentration in water louse tissues reached comparatively higher levels and peaked after ~ 6 days. Neither stickleback nor water louse took up ritalinic acid in tissues despite being present in the water column. Our findings provide valuable data for use in future risk assessment of methylphenidate and will aid in the design of studies aimed at measuring any ecotoxicological effects on, for example, the behaviour or physiology of aquatic organisms.

Pharmaceuticals entering aquatic ecosystems via wastewater effluents are of increasing concern for wild animals. Because some pharmaceuticals are designed to modulate human behaviour, measuring the impacts of exposure to pharmaceuticals on fish behaviour has become a valuable endpoint. While laboratory studies have shown that pharmaceuticals can affect fish behaviour, there is a lack of understanding if behaviour is similarly affected in natural environments. Here, we exposed sea trout (Salmo trutta) smolts to two concentrations of two pharmaceutical pollutants often detected in surface waters: temazepam (a benzodiazepine, anxiolytic) or irbesartan (an angiotensin II receptor blocker, anti-hypertensive). We tested the hypothesis that changes to behavioural traits (anxiety and activity) measured in laboratory trials following exposure are predictive of behaviour in the natural environment (downstream migration). Measures of anxiety and activity in the laboratory assay did not vary with temazepam treatment, but temazepam-exposed fish began migrating faster in the field. Activity in the laboratory assay did predict overall migration speed in the field. In contrast to temazepam, we found that irbesartan exposure did not affect behaviour in the laboratory, field, or the relationship between the two endpoints. However, irbesartan was also not readily taken up into fish tissue (i.e. below detection levels in the muscle tissue), while temazepam bioconcentrated (bioconcentration factor 7.68) rapidly (t 1/2 < 24 h). Our findings add to a growing literature showing that benzodiazepine pollutants can modulate fish behaviour and that laboratory assays may be less sensitive at detecting the effects of pollutants compared to measuring effects in natural settings. Therefore, we underscore the importance of measuring behavioural effects in the natural environment., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Round goby Neogobius melanostomus sagittal (saccular) otolith morphology was compared between males of the two alternative reproductive tactics (termed guarder and sneaker males) and between males captured from sites of high or low contamination. Otolith size increased with fish size and also displayed an ontogenetic shift in shape, becoming relatively taller as otoliths grew in size. Despite a considerable overlap in age between males adopting the two reproductive tactics, size-at-age measurements revealed that guarder males are significantly larger than sneakers at any given age and that they invest more into somatic growth than sneaker males. Controlling for body size, sneaker males possessed heavier sagittal otoliths than guarder males. Subtle otolith shape differences were also found between the two male tactics and between sites of high and low contaminant exposure. Sneaker males had relatively shorter otoliths with more pronounced notching than guarder males. Fish captured at sites of high contamination had otoliths showing slower growth rates in relation to body size and their shapes had more pronounced caudal points and ventral protrusions when compared with fish captured at sites of low contamination. The results are discussed in relation to life-history tradeoffs between the male tactics in terms of reproductive and somatic investment as well as the putative metabolic costs of exposure to contaminants. Overall, this study reveals that male alternative reproductive tactics and environmental contaminants can have small, yet measurable, effects on otolith morphology and these factors should be accounted for in future research when possible., (© 2018 The Fisheries Society of the British Isles.)

Chemical contaminants (e.g. metals, pesticides, pharmaceuticals) are changing ecosystems via effects on wildlife. Indeed, recent work explicitly performed under environmentally realistic conditions reveals that chemical contaminants can have both direct and indirect effects at multiple levels of organization by influencing animal behaviour. Altered behaviour reflects multiple physiological changes and links individual- to population-level processes, thereby representing a sensitive tool for holistically assessing impacts of environmentally relevant contaminant concentrations. Here, we show that even if direct effects of contaminants on behavioural responses are reasonably well documented, there are significant knowledge gaps in understanding both the plasticity (i.e. individual variation) and evolution of contaminant-induced behavioural changes. We explore implications of multi-level processes by developing a conceptual framework that integrates direct and indirect effects on behaviour under environmentally realistic contexts. Our framework illustrates how sublethal behavioural effects of contaminants can be both negative and positive, varying dynamically within the same individuals and populations. This is because linkages within communities will act indirectly to alter and even magnify contaminant-induced effects. Given the increasing pressure on wildlife and ecosystems from chemical pollution, we argue there is a need to incorporate existing knowledge in ecology and evolution to improve ecological hazard and risk assessments., (© 2018 The Author(s).)

Municipal wastewater effluent is a major source of aquatic pollution and has potential to impact cellular energy metabolism. However, it is poorly understood whether wastewater exposure impacts whole-animal metabolism and whether this can be accommodated with adjustments in respiratory physiology. We caged bluegill sunfish (Lepomis macrochirus) for 21 days at two sites downstream (either 50 or 830 m) from a wastewater treatment plant (WWTP). Survival was reduced in fish caged at both downstream sites compared to an uncontaminated reference site. Standard rates of O 2 consumption increased in fish at contaminated sites, reflecting a metabolic cost of wastewater exposure. Several physiological adjustments accompanied this metabolic cost, including an expansion of the gill surface area available for gas exchange (reduced interlamellar cell mass), a decreased blood-O 2 affinity (which likely facilitates O 2 unloading at respiring tissues), increased respiratory capacities for oxidative phosphorylation in isolated liver mitochondria (supported by increased succinate dehydrogenase, but not citrate synthase, activity), and decreased mitochondrial emission of reactive oxygen species (ROS). We conclude that exposure to wastewater effluent invokes a metabolic cost that leads to compensatory respiratory improvements in O 2 uptake, delivery, and utilization.

Pharmaceutical active compounds (PhACs) are increasingly being reported in wastewater effluents and surface waters around the world. The presence of these products, designed to modulate human physiology and behaviour, has created concern over whether PhACs similarly affect aquatic organisms. Though laboratory studies are beginning to address the effects of individual PhACs on fish behaviour, few studies have assessed the effects of exposure to complex, realistic wastewater effluents on fish behaviour. In this study, we exposed a wild, invasive fish species-the round goby (Neogobius melanostomus)-to treated wastewater effluent (0%, 50% or 100% effluent dilutions) for 28days. We then determined the impact of exposure on fish aggression, an important behaviour for territory acquisition and defense. We found that exposure to 100% wastewater effluent reduced the number of aggressive acts that round goby performed. We complimented our behavioural assay with measures of pharmaceutical uptake in fish tissues. We detected 11 of 93 pharmaceutical compounds that we tested for in round goby tissues, and we found that concentration was greatest in the brain followed by plasma, then gonads, then liver, and muscle. Fish exposed to 50% and 100% effluent had higher tissue concentrations of pharmaceuticals and concentrated a greater number of pharmaceutical compounds compare to control fish exposed to no (0%) effluent. Exposed fish also showed increased ethoxyresorufin-O-deethylase (EROD) activity in liver tissue, suggesting that fish were exposed to planar halogenated/polycyclic aromatic hydrocarbons (PHHs/PAHs) in the wastewater effluent. Our findings suggest that fish in effluent-dominated systems may have altered behaviours and greater tissue concentration of PhACs. Moreover, our results underscore the importance of characterizing exposure to multiple pollutants, and support using behaviour as a sensitive tool for assessing animal responses to complex contaminant mixtures, like wastewater effluent., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Pharmaceuticals and personal care products (PPCPs) have been found in wastewater treatment plant (WWTP) effluents and their recipient watersheds. To assess the potential of WWTP effluents to alter fish behaviour, we caged male goldfish (Carassius auratus) for 21-days at three sites along a contamination gradient downstream from a WWTP which discharges into Cootes Paradise Marsh, on the western tip of Lake Ontario. We also included a fourth caging site as an external reference site within Lake Ontario at the Jordan Harbour Conservation Area. We then measured concentrations of PPCPs and monoamine neurotransmitters in caged goldfish plasma, and conducted behavioural assays measuring activity, startle response, and feeding. We detected fifteen different PPCPs in goldfish plasma including six serotonin reuptake inhibitors (amitriptyline, citalopram, fluoxetine/norfluoxetine, sertraline, venlafaxine, and diphenhydramine). Plasma concentrations of serotonin were significantly greater in plasma of fish caged closer to the WWTP effluent outfall site. The fish caged near and downstream of the WWTP effluent were bolder, more exploratory, and more active overall than fish caged at the reference site. Taken together, our results suggest that fish downstream of WWTPs are accumulating PPCPs at levels sufficient to alter neurotransmitter concentrations and to also impair ecologically-relevant behaviours.

Population growth has led to increased global discharges of wastewater. Contaminants that are not fully removed during wastewater treatment, such as pharmaceuticals and personal care products (PPCPs), may negatively affect aquatic ecosystems. PPCPs can bioaccumulate causing adverse health effects and behavioural changes in exposed fish. To assess the impact of PPCPs on wild fish, and to assess whether caged fish could be used as a surrogate for resident wild fish in future monitoring, we caged goldfish in a marsh affected by discharges of wastewater effluents (Cootes Paradise, Lake Ontario, Canada). We collected plasma from resident wild goldfish, and from goldfish that we caged in the marsh for three weeks. We analyzed the plasma proteome and metabolome of both wild and caged fish. We also compared proteomic and metabolic responses in caged and wild fish from the marsh to fish caged at a reference site (Jordan Harbour Conservation Area). We identified significant changes in expression of over 250 molecules that were related to liver necrosis, accumulation and synthesis of lipids, synthesis of cyclic AMP, and the quantity of intracellular calcium in fish from the wastewater affected marsh. Our results suggest that PPCPs could be affecting the health of wild fish populations.

Fluoxetine (Prozac™) is designed to alter human behaviour; however, because many physiological pathways are conserved across vertebrates, this drug may affect the behaviour of fish living in fluoxetine-polluted environments. Although a number of studies have used behaviour to document the sub-lethal effects of fluoxetine, the repeatability of these effects across experiments, across behavioural contexts, and over different exposure durations are rarely considered. Here, we conducted two experiments and assessed how fluoxetine exposure affected a range of fitness-related behaviours in wild round goby (Neogobius melanostomus). We found that fluoxetine impacts round goby behaviour at high (40 μg/l) doses, but not at environmentally relevant low doses (1 μg/l). In both experiments, an acute 3-day exposure to fluoxetine reduced round goby aggression in multiple behavioural contexts, but had no detectable effect on overall activity or social affiliative behaviour. While a chronic 28-day exposure to fluoxetine exposure still reduced aggression, this reduction was only detectable in one behavioural context. Our findings demonstrate the importance of repeated behavioural testing (both between and within experiments) and contribute to a growing body of literature evaluating the effects of fluoxetine and other pharmaceuticals on animal behaviour., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Treated effluents from wastewater treatment plants (WWTP) are a significant source of anthropogenic contaminants, such as pharmaceuticals, in the aquatic environment. Although our understanding of how wastewater effluent impacts fish reproduction is growing, we know very little about how effluent affects non-reproductive physiology and behaviours associated with fitness (such as aggression and activity). To better understand how fish cope with chronic exposure to wastewater effluent in the wild, we caged round goby (Neogobius melanostomus) for three weeks at different distances from a wastewater outflow. We evaluated the effects of this exposure on fish survival, behaviour, metabolism, and respiratory traits. Fish caged inside the WWTP and close to the outfall experienced higher mortality than fish from the reference site. Interestingly, those fish that survived the exposure performed similarly to fish caged at the reference site in tests of aggressive behaviour, startle-responses, and dispersal. Moreover, the fish near WWTP outflow displayed similar resting metabolism (O 2 consumption rates), hypoxia tolerance, haemoglobin concentration, haematocrit, and blood-oxygen binding affinities as the fish from the more distant reference site. We discuss our findings in relation to exposure site water quality, concentrations of pharmaceutical and personal care product pollutants, and our test species tolerance., (Copyright © 2016 Elsevier B.V. All rights reserved.)

To gain a deeper understanding of how environmental conditions affect brain plasticity, brain size was explored across different seasons using the invasive round goby Neogobius melanostomus. The results show that N. melanostomus had heavier telencephalon in the spring compared to the autumn across the two years of study. Furthermore, fish in reproductive condition had heavier telencephala, indicating that tissue investment and brain plasticity may be related to reproductive needs in N. melanostomus., (© 2014 The Fisheries Society of the British Isles.)