Erin S. McCallum

Early-life stress affects the behavioural and neural response of female song sparrows to conspecific song

In songbirds, both song production and song preferences may be influenced by early-life experience. Early-life stress impairs development of the song-control brain regions and permanently affects male song production. However, few studies have examined the effects of early-life stress on female song preferences or the brains of female songbirds. We exposed female song sparrows, Melospiza melodia, to stressors (food restriction or exogenous corticosterone treatment) early in development. When birds were adults, we determined the effects of the stressors on: (1) the behavioural response to high-complexity versus low complexity-songs as well as to conspecific versus heterospecific songs; (2) the volume of song-control brain regions; and (3) the immediate early gene (Zenk) response in auditory forebrain regions following exposure to either conspecific or heterospecific song. We found no significant effect of developmental stress on the strength of the behavioural response to high- versus low-complexity song, but control females showed more selectivity in their behavioural response to conspecific versus heterospecific song when compared with food-restricted or corticosterone-treated birds. We observed no effect of either stressor on volume of the song-control nuclei. Control females exposed to conspecific song had more Zenk-immunoreactive cells in the auditory brain regions than control females exposed to heterospecific song. In contrast, food-restricted and corticosterone-treated females did not have more Zenk-immunoreactive cells after exposure to conspecific song. These results show that stressors known to affect male song production might also affect neural processing of song by females, and their subsequent behavioural response to song.

Exposure to wastewater effluent affects fish behaviour and tissue-specific uptake of pharmaceuticals

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.

In situ exposure to wastewater effluent reduces survival but has little effect on the behaviour or physiology of an invasive Great Lakes fish

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 (O2 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.

Aggression and sociality: conflicting or complementary traits of a successful invader?

Invasion biology research has identified two juxtaposing behavioural traits, aggressiveness and sociality, that may both increase the success of species invasions. Highly aggressive invaders can out-compete native species for resources, while social gregarious invaders can tolerate high conspecific density. In order to tease apart the effects of aggressive versus social tendencies on the success of invasive species, we studied round goby (Neogobius melanostomus), a highly successful invasive fish species now common in the Laurentian Great Lakes. While round goby are well known for being aggressive, much less is known about their tendency to affiliate with conspecifics, in spite of the fact that they thrive in extremely high densities in many of their invaded habitats. We collected round goby from Hamilton Harbour, ON, Canada and conducted three separate experiments to explore group-forming behaviour by measuring preference for conspecifics. We found that round goby have a strong preference to associate with a single conspecific, and that both males and females showed this preference. No overall preference was detected for large versus small groups of conspecifics. Females chose the safety of a shelter over associating with a conspecific but males were equally attracted to conspecifics as shelter. Our results provide new insight into how interactions between aggressive and social behaviours play a role in the rapid spread of invasive round goby.

Seasonal plasticity in telencephalon mass of a benthic fish.

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.

Direct and indirect effects of chemical contaminants on the behaviour, ecology and evolution of wildlife

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.

Altered expression of metabolites and proteins in wild and caged fish exposed to wastewater effluents in situ

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.

Reduced anxiety is associated with the accumulation of six serotonin reuptake inhibitors in wastewater treatment effluent exposed goldfish Carassius auratus

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.

Metabolic Costs of Exposure to Wastewater Effluent Lead to Compensatory Adjustments in Respiratory Physiology in Bluegill Sunfish

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 O2 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-O2 affinity (which likely facilitates O2 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 O2 uptake, delivery, and utilization.

Diet and foraging of Round Goby (Neogobius melanostomus) in a contaminated harbour

Anthropogenic pollution and the introduction of invasive species are two contributing factors to ecosystem degradation. Although Hamilton Harbour (Ontario, Canada), a highly impacted ecosystem, is well-studied, the diet, trophic position, and foraging behaviour of the invasive Round Goby (Neogobius melanostomus) in this area is not well understood. In this study, we compared digestive tract contents, foraging behaviour, and stable isotope values of Round Goby from sites of low and high sediment contamination in Hamilton Harbour. We also assessed prey availability by conducting sediment invertebrate abundance analyses at these sites. Regardless of site, Chironomids, Cladocerans, Copepods and Dreissenids were the most common food items found in Round Goby digestive tracts, and females always had heavier gut contents compared to males. Fish from the high contamination site consumed fewer prey items, had lower gut fullness scores, and fed at a lower trophic level based on lower δ13C and δ15N values. Our results suggest that Round Goby living in highly contaminated areas are feeding less than Round Goby from areas of lower contamination, but that these diet differences do not reflect differences in prey availability. Fish from the high contamination site also typically moved more slowly while foraging. Taken together, these results provide an analysis of the main prey items of Round Goby in Hamilton Harbour, and demonstrate how polluted environments can impact diet, trophic position, and foraging of an introduced fish species.