Physiological Stress Integrates Resistance to Rattlesnake Venom and the Onset of Risky Foraging in California Ground Squirrels.

Using venom for predation often leads to the evolution of resistance in prey. Understanding individual variation in venom resistance is key to unlocking basic mechanisms by which antagonistic coevolution can sustain variation in traits under selection. For prey, the opposing challenges of predator avoidance and resource acquisition often lead to correlated levels of risk and reward, which in turn can favor suites of integrated morphological, physiological and behavioral traits. We investigate the relationship between risk-sensitive behaviors, physiological resistance to rattlesnake venom, and stress in a population of California ground squirrels. For the same individuals, we quantified foraging decisions in the presence of snake predators, fecal corticosterone metabolites (a measure of "stress"), and blood serum inhibition of venom enzymatic activity (a measure of venom resistance). Individual responses to snakes were repeatable for three measures of risk-sensitive behavior, indicating that some individuals were consistently risk-averse whereas others were risk tolerant. Venom resistance was lower in squirrels with higher glucocorticoid levels and poorer body condition. Whereas resistance failed to predict proximity to and interactions with snake predators, individuals with higher glucocorticoid levels and in lower body condition waited the longest to feed when near a snake. We compared alternative structural equation models to evaluate alternative hypotheses for the relationships among stress, venom resistance, and behavior. We found support for stress as a shared physiological correlate that independently lowers venom resistance and leads to squirrels that wait longer to feed in the presence of a snake, whereas we did not find evidence that resistance directly facilitates latency to forage. Our findings suggest that stress may help less-resistant squirrels avoid a deadly snakebite, but also reduces feeding opportunities. The combined lethal and non-lethal effects of stressors in predator-prey interactions simultaneously impact multiple key traits in this system, making environmental stress a potential contributor to geographic variation in trait expression of toxic predators and resistant prey.

Interspecific competition affects the expression of personality-traits in natural populations.

Competition between animal species can cause niche partitioning and shape an individual's phenotype, including its behaviour. However, little is known about effects of interspecific competition on personality, the among-individual variation in behaviour that is consistent across different spatial and temporal contexts. We investigated whether alien grey squirrels (Sciurus carolinensis) influenced the expression of personality traits in native red squirrels (Sciurus vulgaris). In Italy, alien grey squirrels replaced native reds through competition for food resources and space, reducing breeding and recruitment in the native species. We compared personality of red squirrels in red-only (no interspecific competition) and red-grey (with interspecific competition) sites, using arena-tests. The trait activity was measured by Open Field Test while sociability and avoidance were quantified by Mirror Image Stimulation test. Red squirrels co-occurring with the alien species had higher sociability scores and higher between-individual variation in sociability than in red-only sites. Differences in activity and avoidance were not significant. Personality - fitness relationships were not affected by presence or absence of grey squirrels, suggesting that the expression of sociability in red squirrels was not due to short-term selection, but was likely the result of context-related advantages when co-occurring with the competing species.

Social effects of territorial neighbours on the timing of spring breeding in North American red squirrels.

Organisms can affect one another's phenotypes when they socially interact. Indirect genetic effects occur when an individual's phenotype is affected by genes expressed in another individual. These heritable effects can enhance or reduce adaptive potential, thereby accelerating or reversing evolutionary change. Quantifying these social effects is therefore crucial for our understanding of evolution, yet estimates of indirect genetic effects in wild animals are limited to dyadic interactions. We estimated indirect phenotypic and genetic effects, and their covariance with direct effects, for the date of spring breeding in North American red squirrels (Tamiasciurus hudsonicus) living in an array of territories of varying spatial proximity. Additionally, we estimated indirect effects and the strength of selection at low and high population densities. Social effects of neighbours on the date of spring breeding were different from zero at high population densities but not at low population densities. Indirect phenotypic effects accounted for a larger amount of variation in the date of breeding than differences attributable to the among-individual variance, suggesting social interactions are important for determining breeding dates. The genetic component to these indirect effects was however not statistically significant. We therefore showcase a powerful and flexible method that will allow researchers working in organisms with a range of social systems to estimate indirect phenotypic and genetic effects, and demonstrate the degree to which social interactions can influence phenotypes, even in a solitary species.

How to stay perfect: the role of memory and behavioural traits in an experienced problem and a similar problem.

When animals encounter a task they have solved previously, or the same problem appears in a different apparatus, how does memory, alongside behavioural traits such as persistence, selectivity and flexibility, enhance problem-solving efficiency? We examined this question by first presenting grey squirrels with a puzzle 22 months after their last experience of it (the recall task). Squirrels were then given the same problem presented in a physically different apparatus (the generalisation task) to test whether they would apply the previously learnt tactics to solve the same problem but in a different apparatus. The mean latency to success in the first trial of the recall task was significantly different from the first exposure but not different from the last exposure of the original task, showing retention of the task. A neophobia test in the generalisation task suggested squirrels perceived the different apparatus as a different problem, but they quickly came to apply the same effective tactics as before to solve the task. Greater selectivity (the proportion of effective behaviours) and flexibility (the rate of switching between tactics) both enhanced efficiency in the recall task, but only selectivity enhanced efficiency in the generalisation task. These results support the interaction between memory and behavioural traits in problem-solving, in particular memory of task-specific tactics that could enhance efficiency. Squirrels remembered and emitted task-effective tactics more than ineffective tactics. As a result, they consistently changed from ineffective to effective behaviours after failed attempts at problem-solving.

Effects of visitor numbers on captive European red squirrels (Sciurus vulgaris) and impacts on visitor experience.

Visitors to zoological collections can have substantial effects on captive animals that vary according to species, enclosure design, visitor proximity, and husbandry methods. One particularly intense form of visitor interaction occurs in immersive exhibits such as walk-through enclosures. Such enclosures are increasingly common but effects on animal behavior are currently understudied. Here, the behavior of captive European red squirrels (Sciurus vulgaris) is studied in relation to visitor numbers in a walk-through enclosure. We also quantify the correlation between squirrel encounters and visitor experience. Interaction with humans increased significantly as the number of visitors inside the enclosure increased. The number of children present significantly increased locomotion and decreased eating, possibly due to disturbance and squirrels moving away from busy areas. By contrast, the number of adults significantly increased eating and decreased inactivity due to squirrels approaching visitors. The positive reinforcement training used by the keepers (offering food rewards to the squirrels for coming to them to allow routine medical checks) meant that squirrels associated adults with food opportunities. Squirrel encounter rate (number of squirrels seen by each group of visitors) was significantly affected by the number of adults and visitor duration (positive relationships) and noise as perceived by visitors (negative relationship). Encounter rate was positively correlated with overall visitor experience. Our results indicate that visitors affect behavior but this effect is influenced by husbandry methods. It is vital that visitors, especially children, minimize noise, and move slowly in the enclosure, both for the sake of the animals and their own experience.

One plus one: Binary alarm calls retain individual signature for longer periods than single-note alarms in the European ground squirrel (Spermophilus citellus).

Ground squirrels emit species-specific alarm calls that, among other characteristics, differ by the number of elements. Unlike some species that produce single-element calls, e.g., the Speckled ground squirrel (Spermophilus suslicus), individual European ground squirrels (S. citellus) frequently emit binary-element calls in addition to single-element calls. We tested the hypothesis that the time stability of individuality encoded in alarm calls might be better retained by complicating their acoustic structure by adding extra elements. In a semi-captive colony of individually marked European ground squirrels, we repeatedly recorded alarm calls that were produced towards a human by 12 adult (2 males and 10 females) live-trapped animals. Repeated recordings occurred within time spans of a few hours, 2days and 1year from the first recording. Our results showed that individual calls were highly similar within recordings, but less similar between recordings separated by time spans. Individual differences were best retained when we used nine acoustic variables from both elements. The differences were worse when we used nine variables from only the first element and worst when we used nine variables from only the second element. These results supported the caller reliability hypothesis for species that produce multiple-note alarms, e.g., the Richardson's ground squirrel (S. richardsonii).

Seasonal programming, not competition or testosterone, drives stress-axis changes in a partially-semelparous mammal.

Animals must make tradeoffs between reproduction and longevity. This is particularly pronounced in male arctic ground squirrels (Urocitellus parryii), that compete aggressively for territories and mates during a three-week breeding season. Breeding males have high rates of severe wounding, high mortality rates, and high free cortisol levels, along with downstream consequences of chronic stress (weight loss, reduced immune function) that appear to contribute to their early death. The elevated cortisol levels are thought to be a result of the intense intrasexual competition. An alternative hypothesis, however, is that the hormonal change is a seasonal adaptation facilitating the tradeoff of immediate competitive advantage at the expense of long-term survival. We tested a two-part hypothesis: first, that elevated free cortisol during the breeding period is a seasonal change that will still occur in the absence of actual competition, and second, that testosterone maintains this increase. We measured plasma cortisol, corticosteroid-binding globulin, and fecal glucocorticoid metabolites in three groups: wild male ground squirrels, captive males prevented from fighting, and captive castrated males. There were no differences amongst these three groups in free and total plasma cortisol, fecal glucocorticoids, or downstream measures of chronic stress. This suggests that high free cortisol and its effects on breeding males are not a consequence of contest competition during the breeding season, but rather a generalized seasonal change. We found no evidence that testosterone plays a role in maintaining elevated free cortisol in arctic ground squirrel males.

Integrating physiology, behavior, and energetics: Biologging in a free-living arctic hibernator.

The use of animal-borne instruments (ABIs), including biologgers and biotransmitters, has played an integral role in advancing our understanding of adjustments made by animals in their physiology and behavior across their annual and daily cycles and in response to weather and environmental change. Here, we review our research employing body temperature (Tb), light, and acceleration biologgers to measure patterns of physiology and behavior of a free-living, semi-fossorial hibernator, the arctic ground squirrel (Urocitellus parryii). We have used these devices to address a variety of physiological, ecological, and evolutionary questions within the fields of hibernation physiology, phenology, behavioral ecology, and chronobiology. We have also combined biologging with other approaches, such as endocrinology and tracking the thermal environment, to provide insights into the physiological mechanisms that underlie fundamental questions in biology including physiological performance trade-offs, timing and functional energetics. Finally, we explore the practical and methodological considerations that need to be addressed in biologging studies of free-living vertebrates and discuss future technological advancements that will increase the power and potential of biologging as a tool for assessing physiological function in dynamic and changing environments.

Inaccessibility of reinforcement increases persistence and signaling behavior in the fox squirrel (Sciurus niger).

Under natural conditions, wild animals encounter situations where previously rewarded actions do not lead to reinforcement. In the laboratory, a surprising omission of reinforcement induces behavioral and emotional responses described as frustration. Frustration can lead to aggressive behaviors and to the persistence of noneffective responses, but it may also lead to new behavioral responses to a problem, a potential adaptation. We assessed the responses to inaccessible reinforcement in free-ranging fox squirrels (Sciurus niger). We trained squirrels to open a box to obtain food reinforcement, a piece of walnut. After 9 training trials, squirrels were tested in 1 of 4 conditions: a control condition with the expected reward, an alternative reinforcement (a piece of dried corn), an empty box, or a locked box. We measured the presence of signals suggesting arousal (e.g., tail flags and tail twitches) and found that squirrels performed fewer of these behaviors in the control condition and increased certain behaviors (tail flags, biting box) in the locked box condition, compared to other experimental conditions. When faced with nonreinforcement, that is, frustration, squirrels increased the number of interactions with the apparatus and spent more time interacting with the apparatus. This study of frustration responses in a free-ranging animal extends the conclusions of captive studies to the field and demonstrates that fox squirrels show short-term negatively valenced responses to the inaccessibility, omission, and change of reinforcement. (PsycINFO Database Record

Social conditioned place preference in the captive ground squirrel (Ictidomys tridecemlineatus): Social reward as a natural phenotype.

Social behaviors of wild animals are often considered within an ultimate framework of adaptive benefits versus survival risks. By contrast, studies of laboratory animals more typically focus on affective aspects of behavioral decisions, whether a rodent derives a rewarding experience from social encounter, and how this experience might be initiated and maintained by neural circuits. Artificial selection and inbreeding have rendered laboratory animals more affiliative and less aggressive than their wild conspecifics, leaving open the possibility that social reward is an artifact of domestication. We compared social behaviors of wild and captive population of juvenile 13-lined ground squirrels (Ictidomys tridecemlineatus), the latter being 2nd- and 3rd-generation descendants of wild individuals. At an age corresponding to emergence from the burrow, postnatal day (PD) 38, captive squirrels engaged in vigorous social approach and play and these juvenile behaviors declined significantly by PD 56. Similarly, young wild squirrels expressed social proximity and play; affiliative interactions declined with summer's progression and were replaced by agonistic chasing behaviors. Social conditioned place preference testing (conditioned PDs 40-50) indicated that adolescent squirrels derived a rewarding experience from social reunion. Our results support the contention that undomesticated rodents have the capacity for social reward and more generally suggest the possibility that positive affective experiences may support group cohesion, social cooperation, and altruism in the wild.

How will the greening of the Arctic affect an important prey species and disturbance agent? Vegetation effects on arctic ground squirrels.

Increases in terrestrial primary productivity across the Arctic and northern alpine ecosystems are leading to altered vegetation composition and stature. Changes in vegetation stature may affect predator-prey interactions via changes in the prey's ability to detect predators, changes in predation pressure, predator identity and predator foraging strategy. Changes in productivity and vegetation composition may also affect herbivores via effects on forage availability and quality. We investigated if height-dependent effects of forage and non-forage vegetation determine burrowing extent and activity of arctic ground squirrels (Urocitellus parryii). We collected data on burrow networks and activity of arctic ground squirrels across long-term vegetation monitoring sites in Denali National Park and Preserve, Alaska. The implications of height-specific cover of potential forage and non-forage vegetation on burrowing behaviour and habitat suitability for arctic ground squirrels were investigated using hierarchical Bayesian modelling. Increased cover of forbs was associated with more burrows and burrow systems, and higher activity of systems, for all forb heights. No other potential forage functional group was related to burrow distribution and activity. In contrast, height-dependent negative effects of non-forage vegetation were observed, with cover over 50-cm height negatively affecting the number of burrows, systems and system activity. Our results demonstrate that increases in vegetation productivity have dual, potentially counteracting effects on arctic ground squirrels via changes in forage and vegetation stature. Importantly, increases in tall-growing woody vegetation (shrubs and trees) have clear negative effects, whereas increases in forb should benefit arctic ground squirrels.

Evolving communicative complexity: insights from rodents and beyond.

Social living goes hand in hand with communication, but the details of this relationship are rarely simple. Complex communication may be described by attributes as diverse as a species' entire repertoire, signallers' individualistic signatures, or complex acoustic phenomena within single calls. Similarly, attributes of social complexity are diverse and may include group size, social role diversity, or networks of interactions and relationships. How these different attributes of social and communicative complexity co-evolve is an active question in behavioural ecology. Sciurid rodents (ground squirrels, prairie dogs and marmots) provide an excellent model system for studying these questions. Sciurid studies have found that demographic role complexity predicts alarm call repertoire size, while social group size predicts alarm call individuality. Along with other taxa, sciurids reveal an important insight: different attributes of sociality are linked to different attributes of communication. By breaking social and communicative complexity down to different attributes, focused studies can better untangle the underlying evolutionary relationships and move us closer to a comprehensive theory of how sociality and communication evolve.

A comparative study of caching and pilfering behavior in two sympatric species, least chipmunks (Tamias minimus) and eastern chipmunks (Tamias striatus).

We captured least chipmunks (Tamias minimus) and eastern chipmunks (T. striatus) from overlapping populations and assessed their comparative success at heterospecific pilfering in a naturalistic laboratory setting. The smaller species (T. minimus) found their competitors' caches more quickly and with less effort. We traced the success of least chipmunks to foraging behavior that targeted the vulnerabilities of eastern chipmunk caches, and a cache placement counterstrategy that protected their own food stores. The value of pilfered caches for least chipmunks was magnified by their smaller body size and the bigger cache size of their larger competitor. We suggest that heterospecific cache pilferage represents an especially lucrative foraging tactic for small foragers.

Grey squirrels (Sciurus carolinensis) show a feature-negative effect specific to social learning.

Previous laboratory studies on social learning suggest that some animals can learn more readily if they first observe a conspecific demonstrator perform the task unsuccessfully and so fail to obtain a food reward than if they observe a successful demonstrator that obtains the food. This effect may indicate a difference in how easily animals are able to associate different outcomes with the conspecific or could simply be the result of having food present in only some of the demonstrations. To investigate we tested a scatter-hoarding mammal, the eastern grey squirrel, on its ability to learn to choose between two pots of food after watching a conspecific remove a nut from one of them on every trial. Squirrels that were rewarded for choosing the opposite pot to the conspecific chose correctly more frequently than squirrels rewarded for choosing the same pot (a feature-negative effect). Another group of squirrels was tested on their ability to choose between the two pots when the rewarded option was indicated by a piece of card. This time, squirrels showed no significant difference in their ability to learn to choose the same or the opposite pot. The results add to anecdotal reports that grey squirrels can learn by observing a conspecific and suggest that even when all subjects are provided with demonstrations with the same content, not all learning occurs equally. Prior experience or expectations of the association between a cue (a conspecific) and food influences what can be learned through observation whilst previously unfamiliar cues (the card) can be associated more readily with any outcome.

A comparative analysis of the categorization of multidimensional stimuli: I. Unidimensional classification does not necessarily imply analytic processing; evidence from pigeons (Columba livia), squirrels (Sciurus carolinensis), and humans (Homo sapiens).

Pigeons (Columba livia), gray squirrels (Sciurus carolinensis), and undergraduates (Homo sapiens) learned discrimination tasks involving multiple mutually redundant dimensions. First, pigeons and undergraduates learned conditional discriminations between stimuli composed of three spatially separated dimensions, after first learning to discriminate the individual elements of the stimuli. When subsequently tested with stimuli in which one of the dimensions took an anomalous value, the majority of both species categorized test stimuli by their overall similarity to training stimuli. However some individuals of both species categorized them according to a single dimension. In a second set of experiments, squirrels, pigeons, and undergraduates learned go/no-go discriminations using multiple simultaneous presentations of stimuli composed of three spatially integrated, highly salient dimensions. The tendency to categorize test stimuli including anomalous dimension values unidimensionally was higher than in the first set of experiments and did not differ significantly between species. The authors conclude that unidimensional categorization of multidimensional stimuli is not diagnostic for analytic cognitive processing, and that any differences between human's and pigeons' behavior in such tasks are not due to special features of avian visual cognition.

Simple and integrated detours: field tests with Columbian ground squirrels.

An internal representation of space offers flexibility to animals during orientation and allows execution of short cuts and detours. We tested the ability of 19 free-ranging Columbian ground squirrels (Spermophilus columbianus) to perform integrated detours that required travelling under- and aboveground. Squirrels were individually tested on their territories (2 tests) and in an arena (7 tests). During tests, animals could reach food by running aboveground and then through tunnels. For the territory tests, natural tunnels were available. For the arena tests, animals used artificial tunnels within a fenced-in part of the meadow. For the last arena test, tubes were placed aboveground replicating the underground structure. In this test animals were asked to make a simple detour, when the full path to the goal was visible. On their territories, 41% of squirrels performed detours. All animals reached the food in the arena. When choosing an arena detour, squirrels based their decision on the proximity of the burrow as well as on whether it led to food. On the last arena test, more squirrels performed correct detours on the first attempt compared to other tests. The results suggest that ground squirrels can perform simple and integrated detours, but animals perform better if the full path is visible.

Prairie dog alarm calls encode labels about predator colors.

Some animals have the cognitive capacity to differentiate between different species of predators and generate different alarm calls in response. However, the presence of any addition information that might be encoded into alarm calls has been largely unexplored. In the present study, three similar-sized human females walked through a Gunnison's prairie dog (Cynomys gunnisoni) colony wearing each of three different-colored shirts: blue, green, and yellow. We recorded the alarm calls and used discriminant function analysis to assess whether the calls for the different-colored shirts were significantly different. The results showed that the alarm calls for the blue and the yellow shirts were significantly different, but the green shirt calls were not significantly different from the calls for the yellow shirt. The colors that were detected, with corresponding encoding into alarm calls, reflect the visual perceptual abilities of the prairie dogs. This study suggests that prairie dogs are able to incorporate labels about the individual characteristics of predators into their alarm calls, and that the complexity of information contained in animal alarm calls may be greater than has been previously believed.

Lactating red squirrels experiencing high heat load occupy less insulated nests.

The heat dissipation limit hypothesis suggests that the capacity for lactating mammals to transfer energy to their offspring through milk may be constrained by limits on heat dissipation, particularly in species that raise offspring in well-insulated nests. We tested a prediction of this hypothesis by evaluating whether lactating free-ranging red squirrels (Tamiasciurus hudsonicus) occupy less insulated nests when experiencing conditions that increase heat load. In support of the hypothesis, when climate normal ambient temperatures were warm, squirrels supporting large litter masses of furred offspring occupied nests of lower insulative value. These results support the heat dissipation limit hypothesis and suggest that free-ranging mammals may select nests based on their insulative value, not only to reduce heat loss in cold conditions but also to dissipate heat during periods of heat stress.

Flexibility of cue use in the fox squirrel (Sciurus niger).

Recent work on captive flying squirrels has demonstrated a novel degree of flexibility in the use of different orientation cues. In the present study, we examine to what extent this flexibility is present in a free-ranging population of another tree squirrel species, the fox squirrel. We trained squirrels to a rewarded location within a square array of four feeders and then tested them on transformations of the array that either pitted two cue types against one cue type, the majority tests, or all cue types against each other, the forced-hierarchy test. In Experiment 1, squirrels reoriented to the two-cue-type location in all majority tests and to the location indicated by the visual features of the feeders in the forced-hierarchy test. This preference for visual features runs contrary to previous studies that report the use of spatial cues over visual features in food-storing species. In Experiments 2-5 we tested squirrels with different trial orders (Experiments 2 and 3), a different apparatus (Experiment 4) and at different times of the year (Experiment 5) to determine why these squirrels had chosen to orient using visual features in the first experiment. Like captive flying squirrels, free-ranging fox squirrels showed a large degree of flexibility in their use of cues. Furthermore, their cue use appeared to be sensitive both to changes in the test apparatus and the season in which we tested. Altogether our results suggest that the study of free-ranging animals over a variety of conditions is necessary for understanding spatial cognition.

The interaction between personality, offspring fitness and food abundance in North American red squirrels.

Animal personality is now frequently reported in wild and captive populations. It has been shown to be moderately heritable and to have potentially important fitness consequences. Variation in personality within a population may be maintained by balancing selection if different values of personality traits are favoured under different conditions. We measured personality in 98 female North American red squirrels (Tamiasciurus hudsonicus Erxleben), and examined whether its variation could be maintained by changing selection pressures acting via reproductive traits and yearly variation in food abundance. There was no effect of personality on parturition date or litter size, but a female's activity was correlated to the growth rate of her offspring in the nest, and her aggressiveness was correlated to their survival in the nest and overwinter. The magnitude and direction of the effects changed among life history stages and years, possibly in association with food supply in some cases, and may indicate a role for balancing selection in the maintenance of personality.