Ponds as experimental arenas for studying animal movement: current research and future prospects

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

Cross-Latitude Behavioural Axis in an Adult Damselfly Calopteryx splendens (Harris, 1780)

Simple Summary Animals adapt to the environment they live in. If the environment changes, animals usually adapt behaviourally as a first response. By studying behavioural profiles across long distances, we can detect environmental change reflected in shifts in behavioural profiles. This study examined variation in three behavioural axes: activity, courtship and boldness, and the association between these behaviours, i.e., behavioural syndromes, across three damselfly populations along a latitudinal gradient (i.e., climatic gradient). Our study organism was the temperate damselfly Calopteryx splendens. We predicted that behavioural expressions would gradually increase from southern to northern regions. This is because northern animals should compensate behaviourally for a brief and cold breeding season (i.e., time constraint). Activity was the only behaviour feature positively associated with latitudinal gradient. Courtship effort was highest in the central region, whereas boldness values were highest in the north but did not differ between central and south. In the southern region, an activity-boldness and a courtship-boldness syndrome were present, and in the northern region, only an activity-boldness syndrome was found. Our results confirm that environmental variability in biotic and abiotic factors across studied latitudes generates regional differences in behavioural profiles, which do not always follow latitudinal gradient. Behavioural variation is important for evolutionary and ecological processes, but can also be useful when predicting consequences of climate change and effects on species ranges. Latitudinal differences in behaviour have received relatively limited research interest when compared to morphological, life history and physiological traits. This study examined differences in expression of three behavioural axes: activity, courtship and boldness, and their correlations, along a European latitudinal gradient spanning ca. 1500 km. The study organism was the temperate damselfly Calopteryx splendens (Harris). We predicted that the expression of both behavioural traits and behavioural syndromes would be positively correlated to latitude, with the lowest values in the southern populations, followed by central and the highest in the north, because animals usually compensate behaviourally for increasing time constraints and declining environmental conditions. We found that behavioural expression varied along the latitudinal cline, although not always in the predicted direction. Activity was the only behaviour that followed our prediction and gradually increased northward. Whereas no south-to-north gradient was seen in any of the behavioural syndromes. The results, particularly for activity, suggest that climatic differences across latitudes change behavioural profiles. However, for other traits such as courtship and boldness, local factors might invoke stronger selection pressures, disrupting the predicted latitudinal pattern

Uptake, depuration, and behavioural effects of oxazepam on activity and foraging in a tropical snail (Melanoides tuberculata)

Pharmaceuticals are increasingly being detected in surface waters around the globe, giving rise to concerns that they may alter the physiology and behaviour of aquatic organisms exposed in the wild. Invertebrates represent important components of many ecosystems and bear a high potential for transmitting pharmaceutical contaminants to higher trophic levels. Here, we present a laboratory study in which we exposed a freshwater tropical snail, Melanoides tuberculata, to a serial dilution of the benzodiazepine oxazepam ranging from 50 ng/L to 5 mg/L. We tested for subsequent behavioural effects, including locomotor activity and foraging propensity, at two diurnal time points (day and night), and across three days. We found that the snails displayed a high level of behavioural tolerance to all treatments of oxazepam except at the highest exposure, where locomotor and foraging activity declined. We also detected a weak non-monotonic response curve suggestive of behavioural disinhibition at moderate exposure levels. Regardless of treatment, the snails were also less active after three days of exposure and more active during nighttime observations. We measured the uptake of oxazepam in tissues across treatments, showing that it bioconcentrated at up to 29 times the water exposure level (BCF range: 7 - 29). Finally, we characterized the uptake/depuration pharmacokinetics of oxazepam in snail tissues across time, which revealed that the snails reach a steady state equilibrium i

Adult insect personality in the wild-Calopteryx splendens as a model for field studies

Animal personality has received increasing interest and acknowledgment within ecological research over the past two decades. However, some areas are still poorly studied and need to be developed. For instance, field studies focused on invertebrates are currently highly underrepresented in the literature. More studies including a wider variety of traits measured and species tested are needed to improve our understanding of trait-correlation patterns and generalities. We studied nine behavioral traits, in the damselfly Calopteryx splendens, from an array of three experiments: (i) courtship, (ii) aggressiveness, and (iii) boldness, and calculated their repeatability. The behaviors were measured twice in two different contexts: (i) undisturbed territory and (ii) partially deteriorated territory. Traits related to courtship and boldness were all repeatable across the two contexts. Among aggressive behaviors, only one trait (number of hits) was repeatable. This work demonstrates, for the first time, the presence of within-population personality differences in an adult damselfly in the wild. We further propose C. splendens as a promising model species for testing personality in the wild under highly controlled environmental conditions

Multiple Origins of Elytral Reticulation Modifications in the West Palearctic Agabus bipustulatus Complex (Coleoptera, Dytiscidae)

The Agabus bipustulatus complex includes one of Europe's most widely distributed and common diving beetles. This complex, which is known for its large morphological variation, has a complex demographic and altitudinal variation in elytral reticulation. The various depth of the reticulation imprint, both in smaller and larger meshes, results in both mat and shiny individuals, as well as intermediate forms. The West Palearctic lowland is inhabited by a sexually dimorphic form, with shiny males and mat females. In mountain regions, shiny individuals of both sexes are found intermixed with mat individuals or in pure populations in central and southern areas, whereas pure populations of mat individuals are exclusively found in the northern region at high altitude. Sexual selection is proposed as a driving force in shaping this variation. However, the occurrence of different types of reticulation in both sexes and disjunct geographical distribution patterns suggest an additional function of the reticulation. Here we investigate the phylogeographical history, genetic structure and reticulation variation of several named forms within the Agabus bipustulatus complex including A. nevadensis. The molecular analyses recognised several well-supported clades within the complex. Several of the named forms had two or more independent origins. Few south European populations were uniform in reticulation patterns, and the males were found to display large variation. Reticulation diversity and population genetic variability were clearly correlated to altitude, but no genetic differences were detected among populations with mixed or homogenous forms. Observed reduction in secondary reticulation in female and increased variance in male at high altitude in South Europe may be explained by the occurrence of an additional selective force, beside sexual selection. The combined effect of these selective processes is here demonstrated in an extreme case to generate isolation barriers between populations at high altitudes. Here we discuss this selective force in relation to thermal selection

Cost-Effective Pharmaceutical Implants in Fish: Validating the Performance of Slow-Release Implants for the Antidepressant Fluoxetine

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 mu g/g), low (25 mu g/g), or control (0 mu g/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;00:1-10. (c) 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC

Environmentally relevant concentration of caffeine-effect on activity and circadian rhythm in wild perch

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 mu g 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

Oxylipins at intermediate larval stages of damselfly Coenagrion hastulatum as biochemical biomarkers for anthropogenic pollution

Aquatic pollution resulting from anthropogenic activities requires adequate environmental monitoring strategies in sentinel organisms. Thus, biochemical biomarkers have been used as early-warning tools of biological effects in aquatic organisms. However, before using these markers for environmental monitoring, knowledge about their developmental variation is vital. In this study, we assessed baseline levels and developmental variations of a group of potential biomarkers, oxylipins, during the lifespan of the Northern damselfly (Coenagrion hastulatum) using liquid chromatography-tandem mass spectrometry. Effects of wastewater exposure on baseline levels were studied in a subset of damselflies to investigate the responsiveness due to anthropogenic pollution. Thirty-eight oxylipins deriving from four polyunsaturated fatty acids via two enzymatic pathways were detected in damselflies at three larval stages and in the adult form. Overall, oxylipin baseline levels showed developmental variation, which was lowest in the intermediate larval stages. Effects of exposure to wastewater effluent on oxylipin baseline levels were dependent on the life stage and were greatest in the early and intermediate larval stages. The study provides first insights into oxylipin profiles of damselflies at different stages of development and their developmental variation. Based on our results, we propose further strategies for incorporating oxylipins in damselfly larvae as biochemical markers for anthropogenic pollution