El precio postulado por la SISS para el Relleno con Material Seleccionado no tiene sustento estadístico al carecer de observaciones suficientes para estimar

200

Do laboratory assays predict behaviour in the wild? A study with pharmaceutical pollutants

E. McCallum, Umea University; A. Sundelin, J. Fick, Umea University / Department of Chemistry; A. Alanärä, Swedish University of Agricultural Sciences SLU / Department of Wildlife, Fish & Environmental Studies; G. Hellström, Swedish University of Agricultural Sciences SLU; T. Brodin, Umea University / Department of Ecology and Environmental Science

Concern over the impact that pharmaceuticals have on wild aquatic organisms has increased over the past decade. Laboratory studies have shown that pharmaceuticals can cause sub-lethal changes to animal behaviour and physiology; however, few studies have addressed whether effects documented in the laboratory extend to the natural environment. We exposed fish to one of two pharmaceuticals (temazepam and irbesartan) commonly detected in the environment at two doses (80 ng/L, 1500 ng/L; 200 ng/L, 20000 ng/L, for temazepam and irbesartan, respectively). We then assessed how exposure affected fish behaviour in the laboratory (scototaxis to measure anxiety and activity responses) and in the field (downstream dispersal using PIT tags). We found no evidence that either pharmaceutical treatment affected behaviour in the laboratory scototaxis assay. In contrast, fish exposed to both the high and low doses of temazepam dispersed faster downstream when compared to control fish. Irbesartan exposure did not affect fish behaviour in the field. Across all treatments, we also found that activity in the laboratory correlated with migration speed, indicating that fish that were more active in the laboratory also moved faster downstream in the wild. We discuss our findings in relation to differences in tissue bioconcentration for both pharmaceutical compounds in the muscle tissue of fish in our study. Our results emphasize the importance of measuring how pollutants affect ecologically relevant behaviours in the field alongside standard and efficient laboratory assays.

201

Exposure to the widespread androgenic steroid 17β-trenbolone alters behaviour in fish

M.G. Bertram, Monash University / Biological Sciences; M. Saaristo, J.M. Martin, T.E. Ecker, C.P. Johnstone, B.B. Wong, Monash University / School of Biological

Sciences

The capacity of pharmaceutical pollution to alter behaviour in wildlife is of increasing concern to the scientific community. A major pathway of these contaminants into the environment is the treatment of livestock with hormonal growth promotants (HGPs), highly potent veterinary pharmaceuticals that can enter aquatic ecosystems via effluent runoff. Hormonal growth promotants are designed to have biological effects at low doses, often act on physiological pathways that are evolutionarily conserved across taxa, and have repeatedly been detected in ecosystems worldwide. However, despite being shown to cause altered development, reproduction and morphology in various non-target species, relatively little is known about the potential of HGPs to alter ecologically important behaviours, especially across multiple contexts. Here, we investigated the effects of short-term (21-day) exposure to field-detected levels (average measured

concentration: 16 ng/L) of 17β-trenbolone-—a potent growth-promoting veterinary pharmaceutical repeatedly detected in freshwater systems—on a suite of

ecologically important behaviours in female eastern mosquitofish (Gambusia

holbrooki). We found that fish exposed to 17β-trenbolone- were more active and

exploratory in a novel environment (i.e. maze arena), while boldness was not significantly affected. Further, when tested for sociability, exposed fish were again more active and exploratory, and spent less time associating with a shoal of stimulus (i.e. unexposed) conspecific females. Lastly, when assayed for foraging behaviour, exposed fish spent a greater total amount of time within a foraging zone containing of an array of prey items (chironomid larvae) than did unexposed fish, entered this zone more frequently, and were more likely to feed. Further, a significant effect of exposure was detected on the total number of prey items consumed, although treatment-induced increases in foraging behaviour were dependent on female size. Taken together, these findings highlight the potential for sub-lethal levels of veterinary pharmaceuticals detected in the environment to alter sensitive behavioural processes in wildlife across multiple contexts, with possible ecological and evolutionary implications for exposed populations.

202

Selective grazing behaviour of chironomids between three microalgal species under pesticide pressure

J.N. Neury-Ormanni, Irstea / EABX-CARMA; C.N. Doose, INRS - Centre Eau Terre Environnement; B. CHAUMET, Irstea; N. Mazzella, Irstea Bordeaux / UR EABX; N. Majdi, Ecolab / UMR 5245 CNRS; J. Vedrenne, S. Morin, Irstea Bordeaux / UR EABX; W. Traunspurger, Bielefeld University / Animal Ecology The herbicide diuron and the insecticide imidacloprid are amongst the most frequently detected pesticides in French rivers, and each is known to affect many aquatic organisms. However, it is less examined whether and how both pesticides together might induce multistress conditions, which could induce indirect effects such as modification of biological interactions within freshwater microbial communities. This study was undertaken to determine the effect of diuron and imidacloprid alone and in combination on the feeding behavior of chironomid larvae. A first experiment measured the impact of the different contamination conditions at environmental concentrations (5µg L-1 _{for each pesticide) on the} grazing rate of chironomids on three microalgae species, independently. Therefore, two diatom species, Gomphonema gracile (two different morphotype: normal (GG) and teratogen (GT)) and Planothidium lanceolatum (PL), and one green algae

Pseudokirchneriella supcapitata (PS) were offered as food, during 24h. Protein and

lipid contents in microalgae were analysed subsequently. Each pesticide condition elicited a different grazing rate in chironomids with regards to algal species and their nutritional quality, with a general preference for Gomphonema gracile with teratogen shape and Pseudokirchneriella supcapitata. In a second experiment (cafeteria), food selectivity of chironomids was determined under similar contamination conditions during 4h: Under diuron, larvae switched equally among microalgae, then were as mobile as in the control without pesticide. However, imidacloprid and the pesticide mixture condition altered chironomids’ movements and grazing behaviour. In these experiments, we highlighted that chironomids feeding behaviour and food preferences are impacted by pesticides. Herbicide and insecticide exposure, alone or in combination, had contrasting effects on grazing, both directly on the larvae or indirectly (food selectivity according to its quality). Our study illustrates the value of considering the impacts of toxicants on target and non-target organisms across trophic levels to improve ecotoxicological risk assessment in an ecosystemic perspective.

203

Environmental levels of anxiolytic pharmaceuticals alter migration of Atlantic salmon in both lab and field

T. Brodin, Umea University / Department of Ecology and Environmental Science; G. Hellström, Swedish University of Agricultural Sciences SLU; J. Klaminder, Umea University / Ecology and Environmental Science; F. Finn, Umea University; A. Lagesson, Umea University / Department of Ecology and Environmental Science; M. Jonsson, Umea University; J. Fick, Umea University / Department of Chemistry

Humans consume more pharmaceuticals than ever and consumption is set to rise. As a consequence, increasing amounts of pharmaceuticals are released into waterways worldwide with virtually no knowledge of how they might affect aquatic ecosystems. Some conspicuous effects of these emerging contaminants are already

evident including the feminization of fish by contraceptive residue. However, recent work suggests that important effects of pharmaceuticals in aquatic environments are much more widespread than currently believed, and that these effects may result in major changes in species interactions, population survival and ecosystem functioning. In several earlier laboratory studies, we have shown that concentrations of pharmaceuticals presently found in waterways alter important behavioural traits in both aquatic macroinvertebrates and fish, and that this in turn affects both feeding efficiency and predation risk. These results suggest that pharmaceutical contamination of aquatic environments may change species interactions, in particular predator-prey interactions, with severe ecosystem-effects as potential consequence. Recently our research focus has turned towards realistic large-scale studies in lakes and rivers using acoustic telemetry to test if findings from the lab also hold in natural settings. Here I present results from one such study comparing effects of environmental levels of the anxiolytic pharmaceutical Oxazepam on migration pattern of Atlantic salmon (Salmo salar) in the lab and the field. In the lab, salmon exposed to the drug migrated approximately twice as fast as unexposed salmon and the subsequent field-study generated similar results, validating the results found in the lab. This pharmaceutically induced change in migration-intensity has the potential to be a key determinant between survival and mortality of salmon individuals and as such important for population persistence as migration intensity is believed to be adapted to the environmental conditions of the river in question. The overall finding of recent studies suggests that effects of pharmaceutical contamination of natural systems might be much more widespread than we predict based on conventional ecotoxicological tests.

204

Can personality influence the response of fish to environmental contaminants?

M. Oliveira, University of Aveiro; M. Sampaio, T. Santos, University of Aveiro / Biology Department & CESAM; A.L. Machado, University of Aveiro / CESAM Department of Biology; M. Barbosa, Department of Biology & CESAM - University of Aveiro / Biology; I. Domingues, University of Aveiro / CESAM Department of Biology

Stress is a determinant factor reducing animal welfare. Currently, it is recognized that animals react differently as a function of their personalities, or stress coping styles (i.e. consistency in behavioural and physiological responses across time or contexts). However, the role of personality in modulating individual response to environmental contaminants have received limited attention, despite the recognition that personality traits associated with a shy-bold continuum play an important role in animal fitness. Knowing that pharmaceuticals can interfere with personality, one question arises: what is the role of personality on animals’ response to contaminants? In this research, zebrafish was sorted according to their exploration of a novel environment into shy and bold individuals, and subsequently exposed during 96h to carbamazepine, a human pharmaceutical, suggested as a marker of anthropogenic pollution. Assessed responses included behaviour (distance swan, position in the tank and time spent swimming) and biochemical markers associated with oxidative stress, neurotransmission and energy metabolism. Overall, our results showed significant differences between control shy and bold organism with behaviour endpoints demonstrating to be very sensitive to stressor conditions. Although carbamazepine alone did not show considerable effects in the assessed endpoints, strong interactions were found between personality and pharmaceuticals, supporting further studies.

205

Effects of fluoxetine on anxiety-related behaviours and physiology in a songbird

S.E. Whitlock, Environment Department, University of York / Environment; R. Shore, Centre for Ecology & Hydrology (NERC); J. Lane, Animal and Plant Health Agency; K. Herborn, Newcastle University / Centre for Behaviour and Evolution; M.G. Pereira, Centre for Ecology & Hydrology / Lancaster; K. Arnold, University of York / Environment

Several species of bird are known to forage directly on invertebrates at wastewater treatment plants (WWTPs), representing an exposure route to a range of

contaminants including pharmaceuticals. The selective serotonin reuptake inhibitor (SSRI) fluoxetine is heavily prescribed and has been widely detected at WWTPs. Since fluoxetine is commonly prescribed for anxiety, we hypothesised that the antidepressant could modulate anxiety behaviour and physiology in exposed birds. Anxiety is an important state which arises in response to a real or perceived threat, enabling the individual to respond appropriately. Contaminants with the potential to alter anxiety-related behaviours are thus of concern to wildlife. We conducted a study to investigate the effects of chronic exposure to an environmentally relevant concentration of fluoxetine in a model songbird, the Eurasian starling (Sturnus

vulgaris). We used a combination of behavioural and physiological endpoints to

assess the effects of fluoxetine, specifically: 1) object neophobia, 2) exploration in a novel environment, 3) activity levels, 4) faecal corticosterone (CORT) metabolite concentration; and 5) leg skin temperature. Compared with pre-treatment data, fluoxetine-treated birds became less neophobic on average after six weeks of dosing, indicating a decrease in anxiety behaviour. There was no such reduction in neophobia in the control group. After six weeks of dosing, control birds became more active on average but fluoxetine-treated birds showed no increase in activity,

44 SETAC Europe 28th Annual Meeting Abstract Book

indicating increased lethargy in the fluoxetine birds relative to controls. There was no clear effect of treatment on exploratory tendency. Finally, infrared

thermography showed that fluoxetine-treated birds had significantly colder legs compared with controls. This indicated that, as observed in humans, fluoxetine causes vasoconstriction, which in birds will affect the ability to thermoregulate. This study provides further evidence that low, environmentally relevant concentrations of pharmaceuticals can cause sublethal changes to behaviour and physiology that are predicted to impair the capacity of wildlife to respond appropriately to environmental changes.

Can trends in wildlife populations revolutionise our