Behavioural Processes

Zoological gardens strive to prioritize excellent animal care and adhere to the highest standards of animal welfare, ensuring that the conditions of animals` social and physical environment are as close as possible to those in the wild. This study investigates the effects of group split and relocation to the new enclosure on the behaviour of western lowland gorillas (Gorilla gorilla gorilla) (N = 7) living in Prague Zoo, Czech Republic. We conducted over 289 hours of behavioural observations focusing on daily activities, social interactions, and behaviours that serve as potential stress and welfare indicators. The group split led to establishment of two groups in two separate enclosures; the old enclosure consisted of a "bachelor group" (i.e. three males) and a new enclosure consisted of a mixed-sex group (i.e. three females and one juvenile male). The behavioural comparisons across different study periods were conducted using linear mixed models (LMMs). The changes led to an increase in time spent moving, feeding, being in social proximity, and higher rates of approaches among the gorillas, as well as to a decrease in rates of self-directed and "undesirable" behaviors. Our findings indicate that the gorillas effectively adapted to the changes, most likely by relying on social support, to navigate new conditions. This study contributes to our understanding of how socio-cognitively complex species cope with necessary alterations in captive animal care programs. Furthermore, these observations may inform strategies to enhance the welfare of zoo-housed animals and to improve their captive care.

The vast majority of birds are diurnal and concentrate their vocal activity during daylight hours. However, some diurnal birds can also be vocally active at night, although the functions of this phenomenon remain poorly understood. We conducted playback experiments in the Warta Landscape Park (central Poland) to determine whether nocturnal singing by diurnal birds serves breeding-related functions by analysing responses to playback of songs from unfamiliar conspecific males. The sedge warbler (Acrocephalus schoenobaenus) was selected as the focal species because it exhibits relatively high levels of nocturnal song activity, while nine additional diurnal species detected near focal sedge warbler territories were included to explore whether responsiveness to nocturnal conspecific song extended across a broader taxonomic range. Playback experiments were conducted during the early and late stages of the breeding season and during the early and late parts of the nautical night. Out of 10 species tested, three responded vocally: sedge warbler, Savi s warbler (Locustella luscinioides), and common snipe (Gallinago gallinago). Sedge warblers did not modify song rate and song duration but increased flight activity after nocturnal playback. Savi s warblers and common snipes produced more vocalisations after playback than before in May, a pattern consistent with territorial defence function. General nocturnal vocal activity was higher at the beginning of the season, suggesting that birds motivation to establish territories and form pairs can extend into the night, providing additional benefits. Moreover, the probability of singing by the sedge warbler was higher in the latter part of the night. Our study demonstrates that nocturnal stimulation of foreign male playback of diurnal birds can elicit vocal responses from conspecifics, suggesting that nocturnal singing can occur in the absence of obvious artificial light pollution, but in the case of some species and environmental conditions, it may contribute to nocturnal social communication, especially in the early breeding season.

Inequity aversion, the sensitivity to inequitable outcomes or processes, has been widely studied in nonhuman primates since 2003. Yet, findings remain debated as some could be explained by alternative mechanisms such as frustration or loss aversion, so-called individual contrasts. Previous work on nonhuman animals has restricted the definition of inequity aversion to the social domain. Here, we propose distinguishing between two different forms: a socially driven form, which depends on comparison with another individual, and an individually based form, caused by a discrepancy between ones own effort and the expected outcome. Using a task-based methodology, we manipulated both effort and the amount of reward to test for the existence of those different forms and distinguish them from individual contrasts. We presented seven Tonkean macaques (Macaca tonkeana) and three brown capuchins (Sapajus apella) with low and high effort tasks associated with low and high reward quantities. To test for the presence of individually based inequity aversion, subjects were tested alone in an individual phase. In some of the trials, they were rewarded less than they deserved for the task. To test for socially based inequity aversion, two individuals performed the same task in a social phase but, in some trials, one received a higher-value reward than the other. We recorded the latency to engage in the task as an indicator of reluctance. In the individual phase, macaques were slower to engage with inequitably rewarded tasks, a pattern consistent with an individually based expectation of equity. By contrast, capuchins were faster in this context, suggesting that their responses were more likely driven by individual contrast effects. In a context of social inequity, both species slowed engagement in tasks with inequitable rewards, suggesting an aversion to socially based inequity. These results demonstrate that such a methodology provides a means to study non-social components of inequity aversion. Future studies, conducted on larger and more diverse samples, with rigorous motivational controls and explicit tests looking at the understanding of the link between task and reward, are needed to confirm whether nonhuman primates can display inequity aversion independently of social comparison.

RationaleAdolescence is a period of heightened vulnerability to nicotine reinforcement. While zebrafish are a valuable model for investigating drug reward, standard conditioned place preference (CPP) assays typically test subjects in isolation. In this highly social species, solitary testing may act as an environmental stressor that confounds behavioral readouts. ObjectivesThis study examined how social context during testing (isolated vs. grouped) affects experimental attrition, behavioral stability, and nicotine CPP expression in late juvenile zebrafish. MethodsZebrafish housed in groups of four were tested either individually (isolated) or in their housing groups (grouped) during daily 20-minute sessions. Following baseline preference assessments, subjects underwent six days of conditioning pairing their initially non-preferred compartment with fish water or nicotine (0.5, 1.6, or 5.0 {micro}mol/L). Place preference, locomotion, and thigmotaxis were assessed on a drug-free test day. ResultsIsolated testing reduced distance traveled, decreased swimming speed, and increased time spent near tank walls, indicating heightened anxiety-like behavior. Experimental attrition was significantly higher in isolated (38.9%) than grouped (2.5%) subjects. Grouped subjects developed significant place preference at 1.6 and 5.0 {micro} mol/L nicotine, whereas preference was not detectable in isolated subjects. ConclusionsSolitary testing acts as a stressor that increases experimental attrition and masks place preference. Conversely, testing in the presence of conspecifics stabilizes behavior and facilitates the detection of nicotine reward in late juvenile zebrafish.

Voluntary physical activity is a highly motivated behavior with important implications for physical and mental health, yet the neural and endocrine mechanisms underlying motivation to exercise remain poorly understood. In contrast, motivation for sugar/palatable foods, drugs, and sex has been extensively characterized using operant paradigms. Here, we describe a novel progressive ratio operant task to measure motivation to run, independent of running ability. Using female Long Evans rats, which exhibit robust voluntary running behavior, we validated this paradigm by applying a manipulation well known to enhance the motivation to run: calorie restriction. Calorie-restricted animals exhibited increased operant responding to gain access to a running wheel, thus demonstrating heightened motivation for exercise. More specifically, calorie-restricted rats completed more ratios, reached a higher breakpoint in the progressive ratio task, ran more, and spent more time in the operant chamber. We did not observe any effects of calorie restriction on the estrous cycle or steroids (e.g. corticosterone, testosterone) in the blood or brain. Importantly, our task dissociates the motivational drive for physical activity from the ability to perform the physical activity itself, providing a new paradigm for studying the neural and endocrine mechanisms that regulate exercise motivation.

Considerable evidence suggests that the motivational control of instrumental action depends on incentive learning; i.e., on the opportunity to learn how the value of the consequences or outcome of an action, (e.g., a specific food) varies under different motivational conditions (e.g., under different degrees of hunger). The current study investigated whether learning the values of high-protein and high-carbohydrate rewards under different degrees of protein and carbohydrate appetite is also necessary for these nutrient-specific appetites to exert control over instrumental performance. Experiment 1 gave differing consummatory experience to whey protein and polycose carbohydrate outcomes under protein and carbohydrate appetite and found that, without the opportunity for incentive learning, the performance of actions earning these outcomes was insensitive to a shift in appetite. However, once the opportunity for incentive learning was provided, the rats increased their instrumental performance on a lever that earned the whey outcome relative to the polycose lever when protein hungry and on the polycose lever relative to the whey lever when carbohydrate hungry. Experiment 2 assessed how these nutrient-specific states exerted this control; whether, once learned, nutrient values were immediately controlled by nutrient appetite or whether this was based on conditional control acquired during experience with the outcomes under different nutrient appetites. We found that exposure to an outcome under a single nutrient-specific state was not sufficient to establish state-specific control. Instead, establishing the conditional control of outcome value required exposure to both the whey and polycose outcomes under both protein and carbohydrate appetites.

The ability of prey to eavesdrop on predator vocalizations is expected to increase survival by reducing detection and capture. Unfortunately, most research has been conducted in vertebrates, and little is known about this ability in invertebrates. We measured latency to emerge, overall activity, and shelter visits in wild-caught fall field crickets (Gryllus pennsylvanicus) in response to acoustic playback. Stimuli included multiple predator vocalizations, non-predator vocalizations, white noise, and a control. We predicted that crickets would reduce activity, spend more time in shelter, and freeze in response to stimuli representing greater risk. Contrary to our predictions, crickets traveled greater distances, spent more time moving, and spent less time in shelter in response to predator vocalizations versus controls. We did not, however, find clear differences in responses between predator vocalizations and other treatments. Our results suggest that crickets may not differentiate between the vocalizations of predators, non-predators, and other abrupt sounds. Consequently, eavesdropping may not be a viable method of assessing predation risk for this species and its general use remains unclear.

Animals experience stressors in a variety of contexts that result in activation of neuroendocrine and cellular stress responses. Release of stress hormones can disrupt or restore redox homeostasis, and the resulting changes in oxidative states, physiology and behavior vary by an individuals stress coping style. However, oxidative stress can also directly modulate neuroendocrine stress signaling. To what extent individual differences in brain antioxidant levels alter behavioral stress levels is not well understood. The present study investigated how N-acetylcysteine amide (NACA), an antioxidant and glutamate-modulating compound, regulates stress behavior across zebrafish (Danio rerio) with different stress coping styles (proactive, reactive). Following 24-hour exposure to NACA or control conditions, we quantified individual and composite stress behaviors using a Light-Dark Test (LDT). As expected, both proactive fish and NACA-treated fish showed significantly lower stress behaviors compared to reactive and control animals, respectively. Notably, stress-reducing effects of NACA were only seen in those with a reactive stress coping style. Overall, our data suggest that antioxidant mechanisms (e.g., glutathione system) may be key in facilitating the distinct behavioral and physiological responses to stressors that characterize alternative stress coping styles. The results underscore how individual differences in stress coping style and redox state can influence behavioral responses to stress.

Behaviour innovation plays a pivotal role in a species adaptability to dynamic environments. Investigating innovative behaviour and its underlying mechanism is therefore crucial for elucidating the development of cognitive flexibility across animals. Animal personality--which shapes how individuals perceive and engage with their surroundings--could offer insights into individual variation in this process. This study used a three-step foraging puzzle to evaluate the innovation capacity in 28 rosy-faced lovebirds (Agapornis roseicollis), specifically examining their capacity to recombine individually innovated component behaviours into integrated, more sophisticated techniques. We found that nearly half of the individuals spontaneously innovated multiple component behaviours to solve novel puzzles. Crucially, when challenged with a more sophisticated task, they recombined these behaviours into functionally dependent sequences without prior social demonstration. We further identified sex, persistence, and asocial learning capacity as key predictors of innovative problem-solving performance, with females, persistent individuals, and superior asocial learners excelled at problem-solving. Our findings demonstrate that behavioural innovation is not a static event, but a dynamic process--modulated by physical, cognitive, and personality variables--in which behaviours are flexibly transferred and recombined into increasingly complex forms to enable rapid individual adaptation.

Although female rats are typically less aggressive than male rats, lactating females will vigorously defend their nests and pups against an intruder. Much attention has been directed at the consequences of this aggression on the intruder and less on the consequences for the mother and her subsequent interactions with her pups. Here, we exposed resident Sprague-Dawley dams to the resident-intruder paradigm twice daily for five consecutive days, beginning when the dam's (RES) pups were 7 days old, to assess social stress effects on maternal behavior and neurobiology. Controls were dams that had time-matched (TMC) separation from their pups but were not exposed to intruders, and naive moms which were never separated nor exposed to an intruder (CTL). We assessed the dam's subsequent behavior and interactions with her pups on Day 1 and Day 5, and Fos expression after Day 5 in select regions of the prefrontal cortex, amygdala, hypothalamus and periaqueductal gray of the midbrain. In separate cohorts, after pups were weaned, the dams underwent restraint stress and plasma corticosterone assayed. PCA analysis of the dam's behaviors identified three components: normal self-focused behaviors; nurturing behaviors and rough non-nurturing behaviors. Relative to CTL, RES dams exhibited more disrupted behaviors towards their pups, including, rough transport, stepping on pups, and flinging/tossing pups around the cage. In contrast, TMC Dams showed some, but fewer changes relative to CTL, suggesting that separation from pups alone does not account for all disrupted behavior in RES dams. The bulk of these behavioral effects occurred in the first 5-10 min after reunion with the pups and were seen on both the first and fifth day of testing. Of the brain regions examined, the prefrontal cortex was activated by both the defeat/intruder stress (RES) and separation stress (TMC), whereas the dorsal PAG was activated specifically by the defeat/intruder stress. The medial and basolateral amygdala exhibited differential neuronal activity between the RES defeat/intruder-exposed dams and the other two groups. The RES moms exhibited an insufficient adrenocortical response to acute restraint stress. The results suggest that amygdala-dPAG activity is important for dissociating disrupted maternal care in RES (due to defense of the nest against an intruder) from simple pup separation, both of which activate the mPFC. The experience of repeatedly defending the nest may induce subsequent disruptions in HPA responses. The amygdala-dPAG pathway may regulate aspects of stress and emotional regulation exhibited by mothers who defend their offspring against intruders.

Healthy lifestyle habits are a key pillar of physical, psychological, and social well-being, yet university years are often marked by declines in sleep quality, physical activity, and other health behaviors that may affect academic performance. This cross-sectional study examined these associations in 390 Spanish university students, assessed via an online and in-person questionnaire covering academic trajectory (grade point average, credits passed, curricular progression), sleep quality (PSQI), physical activity (IPAQ), tobacco use, Body Mass Index (BMI), and sociodemographic data. Bivariate associations were tested with chi-square, Pearson correlation, t-tests, and ANOVA, followed by generalized linear models for significant variables. Sports participation, but not IPAQ-measured physical activity, was significantly associated with academic progression, defined as advancing one grade level per year. Sleep indicators showed mixed, partly counterintuitive associations: poorer nocturnal sleep was linked to better progression, whereas better sleep quality predicted a higher percentage of credits completed. BMI was a modest positive predictor of credit completion, and underweight students performed significantly worse than overweight or obese peers. Tobacco use showed no significant associations. These findings suggest that sleep-related behaviors, sports participation, and, to a lesser extent, BMI, are relevant correlates of academic progress, supporting university health policies addressing sleep, physical activity, and nutrition.

Behavioral test batteries are increasingly used to characterize multiple functional domains in zebrafish, yet the potential impact of test sequence on behavioral outcomes remains poorly defined. Here, we systematically evaluated whether test order influences behavioral responses in a three-assay battery comprising the novel tank test (NTT), mirror-induced aggression (MIA), and social preference (SP) test. Adult zebrafish (Danio rerio) were exposed to all possible permutations of the three assays in a fully counterbalanced design, allowing assessment of order effects across locomotor, anxiety-like, aggression-related, and social behaviors. Test order produced modest and parameter-specific effects, primarily affecting locomotor activity in the NTT and social proximity in the SP assay. Time-course analysis revealed within-test behavioral dynamics, with limited evidence that test order modulates early adaptation or late engagement with the testing environment but does not alter overall temporal response profiles. Sex-dependent effects were assay-specific and most pronounced in the NTT, with no consistent sex differences observed in MIA or SP. To evaluate the global structure of behavioral variation, Principal Component Analysis (PCA) was performed across assays. Despite localized effects of test order, no clear multivariate separation between test sequences was observed, indicating that sequential testing does not produce distinct baseline phenotypes. Together, these findings support the robustness and reproducibility of multidomain behavioral batteries while highlighting the importance of standardized test-order reporting to improve cross-study comparability.

Operant paradigms are powerful tools to quantify motivation and reward. Traditionally, operant conditioning research has been limited to food and drug reinforcers. Recent advances in commercially available operant equipment, however, allow for the quantification of social motivation. These operant assays are an improvement over commonly used social preference tasks, as they enable direct measurement of the effort and motivation driving social behavior. Based on a design by Venniro et al. (2020), the MedPC social operant boxes modify the traditional operant box setup for social interactions. The experimental rat can lever-press to raise a door for an interaction with a target rat behind a porous barrier. These social operant boxes have been widely adapted to test social behavior in adult and adolescent rodents and investigate how a range of conditions (e.g. stress, drug taking, etc.) affect social motivation. However, there is a gap in assessing maternal motivation for pups during the postpartum period, despite ample evidence that postpartum social behavior is highly relevant for offspring health outcomes. Here, we detail 3D-printed modifications to the standard Med PC social operant boxes to adapt the social target chamber to safely house neonatal pups. We have also developed testing protocols to assess motivation during the limited postpartum period. These data demonstrate that, with simple modifications to social operant chambers and testing protocols, the field can implement advanced behavioral approaches to directly assess maternal motivation.

While hamsters are commonly housed in groups within pet shops in Japan, small cages are often thought to restrict their physical activity, leading to arguments that larger cages should be provided. Although previous research has investigated how cage size and running wheel availability influence activity levels in individually housed hamsters, no studies to date have examined these specific effects in a social housing context. Therefore, this study investigated how cage size and the presence of a running wheel affect the activity levels of individual hamsters using the group-housing conditions with five hamsters. Video recordings were captured for 24 hours across four distinct cage environments using a camera installed directly above each cage. From these recordings, the distances traveled on both the cage floor and the running wheel were calculated for each hamster and statistically analyzed. The results revealed that overall activity levels were significantly higher in cages equipped with a running wheel than in those without. Although cage size did not yield a statistically significant difference, a marginal trend toward higher activity in larger cages was observed.

Early life adversity can produce persistent changes during development that increase vulnerability to neuropsychiatric disorders. Although disruptions in reward processing are widely recognized as a hallmark of these disorders, reward is not a single construct. Distinct forms of reward including social interaction and primary rewards such as food rely on overlapping but dissociable neural circuitry and may be differentially affected by adverse experiences. Here, we used a rodent model of neonatal predator odor exposure (POE) to determine how early life threat influences motivation for social and sucrose reward in adulthood using operant procedures. Neonatal POE reduced adult motivation for a social reinforcer during an operant social self-administration task, whereas motivation for a sucrose reinforcer was unchanged. However, we did find a significant difference in sucrose self-administration with POE females pressing more for sucrose than control females. These findings demonstrate that neonatal threat does not produce a generalized deficit in motivation but rather selectively alters motivation across distinct reward domains. Together, this work identifies social reward as a particularly vulnerable behavioral domain following early life threat and provides new insight into how adverse developmental experiences shape adult reward-related behavior.

Long-term memory has been extensively documented in many animals across a range of ecological contexts. Far less is understood, however, about how long memories persist after active modification or how conflicting memories are adaptively regulated to enable flexible behaviour. Parrots are widely recognized for their high intelligence and cognitive capability, yet the study of memory has focused on only a handful of large parrot species. Here, using a binary choice symbolic system, we conducted three experiments on 28 rosy-faced lovebirds (Agapornis roseicollis) to investigate (1) memory retention of initial associative learning at intervals of two weeks, half a year, and one year; (2) reversal learning, where the rewarded and unrewarded symbols were swapped; and (3) memory retention, following reversal learning at two-week and half-year intervals. We revealed that (a) initial associative learning memory was strikingly persistent, remaining detectable after nearly 600 days; (b) reversal learning performance was comparable to that of large parrot species; and (c) while the post-reversal forgetting curve mirrored the pattern of initial learning, it exhibited a shorter retention duration and a lower retention plateau. These findings suggest that forgetting may serve as an adaptive mechanism that regulates the retrievability of conflicting information, rather than a simple erasure of memory traces. When the environment changes, the retrievability of previously functional but now maladaptive memories is suppressed, but their long-term storage is preserved for potential future use under analogous circumstances. Such dynamic regulations prevent the repetitive overwriting of information in a fluctuating environment, thereby fostering greater behavioural flexibility across dynamic scenarios.

Chorus howls play a central role in wolves' (Canis lupus) social communication, shaping both intra- and inter-pack interactions. While synchrony has been shown to influence perceived threat from outgroups in birds and humans, whether the synchrony of an outside pack's chorus howl plays a role in wolf pack behaviour remains unclear. This study investigated behavioural responses of 19 zoo-housed wolf packs to playbacks of synchronous and asynchronous chorus howls from unfamiliar packs, in comparison to a dove control call. We hypothesized that packs would show stronger intra-pack cohesion and greater spatial withdrawal in response to more synchronous, and thus cohesive - and potentially more formidable - sounding vocalisations. Alternatively, asynchronous howls might elicit stronger responses due to their ambiguity and the "Beau Geste" effect (where asynchronous signals make it harder to infer the number of callers). Synchronous howls resulted in more distance from the sound source and less activity compared with asynchronous howls. This could be interpreted as a more cautious response towards a perceived threat. However, since there was no effect on pack-directed behaviour and no difference between the synchronous and control sound, lower interest or motivation to engage with the stimulus cannot be excluded. In contrast, wolves spent more time near the sound-source after hearing asynchronous howls, approached it more, and oriented towards it for longer compared to synchronous playbacks, a pattern consistent with threat-investigation. In how far these reactions result from a more difficult localisation of synchronous howls or more ambiguity in pack size in the asynchronous playbacks, as well as how this translates to wild wolf behaviour, remains to be investigated.

Song evolves rapidly in songbirds, as has been proven for a substantial number of songbird species by demonstrating acoustic differences between current songs and "historical" songs recorded 20 or 30 years previously. In two species of songbirds, white-crowned sparrows and savannah sparrows, it has been further shown that evolutionary changes over such time spans are sufficient to affect the response of receivers, with territorial males responding more aggressively to current songs than to historical ones. These two species, however, have especially low population variability in song, with most males in any population singing the same, single song type; this background of song uniformity makes it especially easy to discern temporal changes. Here we examine response to temporal change in song in a third sparrow species, the song sparrow, in which population variation in song is much higher: males sing 5-13 song types each, with low song-sharing between males, so that hundreds of distinct song types occur in a local population. We find evidence that temporal change has occurred in the songs of our study population, in that 24 song current song types share more introductory phrases with other current song types than do 24 historical song types recorded 27-29 years earlier. Nevertheless, in a song playback experiment, current males showed no difference in response to current and historical songs. The results are in accord with the hypothesis that high levels of population variability in song make temporal changes in song difficult to discern.

One challenge of reintroduction programs is ensuring that captive individuals are prepared for life in the wild, including having the ability to forage on wild resources. Foraging proficiency can be especially challenging to foster in species with a wide dietary breadth or reliance on ephemerally available resources. A relatively unexplored obstacle to release readiness may be an animals hesitancy to approach or consume unfamiliar foods, i.e. their neophobia and dietary conservatism. In this study, we examined the responses of a conservation breeding population of the extinct-in-the-wild alal[a] (Corvus hawaiiensis), a dietary generalist, to novel and familiar native and non-native fruits. Birds were similarly willing to contact both novel and familiar fruits, suggesting little aversion to the presence and appearance of novel food items. However, birds showed a clear preference for contacting non-native fruits over native fruits, regardless of familiarity. Additionally, while birds were willing to contact novel items, they were less likely to consume novel native fruits, suggesting that dietary conservatism may limit incorporation of these resources in the wild. Finally, juvenile birds were less likely to approach the testing setup and contact items, suggesting that neophobia may constrain their exploration of novel environments and resources. We discuss the implications of alal[a] preferences for non-native foods and reluctance to consume novel native items for pre-release preparations and the goal of wild species recovery.

Brydes whales (Balaenoptera edeni edeni) are nationally protected in China, and the waters around Weizhou Island in the Beibu Gulf support one of the countrys few regularly observed coastal groups. However, acoustic data for this population remain limited, and the potential effects of local vessel noise are poorly described. We conducted 16 vessel-based surveys around Weizhou Island and adjacent waters in January 2024 using a low-disturbance sailboat platform and passive acoustic recorders, with concurrent visual observations where possible. We identified 734 low-frequency signals classified as putative Brydes whale vocalizations and quantified their temporal and spectral parameters. Call duration was significantly negatively correlated with maximum frequency and center frequency, but not with minimum frequency or bandwidth. Comparisons with published records indicate that the recorded signals are most similar to vocalizations previously reported from juvenile Brydes whales or mother-calf pairs, although individual source attribution could not be confirmed. Speedboat passage significantly increased root-mean-square sound pressure levels, and the dominant noise band overlapped the frequency range of the recorded Brydes whale signals, indicating potential for acoustic masking. These results expand the bioacoustic baseline for Brydes whales in Chinese coastal waters and provide evidence relevant to the management of vessel activity and whale-watching tourism around Weizhou Island.