Spontaneous tempo production in cockatiels (Nymphicus hollandicus) and jungle crows (Corvus macrorhynchos).

Musical and rhythmical abilities are poorly documented in non-human animals. Most of the existing studies focused on synchronisation performances to external rhythms. In humans, studies demonstrated that rhythmical processing (e. g. rhythm discrimination or synchronisation to external rhythm) is dependent of an individual measure: the individual tempo. It is assessed by asking participants to produce an endogenous isochronous rhythm (known as spontaneous motor tempo) without any specific instructions nor temporal cue. In non-human animal literature, studies describing spontaneous and endogenous production of motor tempo without any temporal clue are rare. This exploratory study aims to describe and compare the spontaneous motor tempo of cockatiels and jungle crows. Data were collected on spontaneous beak drumming behaviours of birds housed in laboratory. Inter beak strokes intervals were calculated from sound tracks of videos. The analyses revealed that inter beak strokes intervals are non-randomly distributed intervals and are isochronous. Recorded spontaneous motor tempos are significantly different among some cockatiels. Since we could only conduct statistical analysis with one corvid, we cannot conclude about this species. Our results suggest that cockatiels and jungle crows have individual tempos, thus encouraging further investigations.

Salivary corticosterone measurement in large-billed crows by enzyme-linked immunosorbent assay.

Salivary corticosteroid measurement, as a surrogate for plasma corticosteroid levels to evaluate an animal's stress or metabolic state, commonly used in mammals. However, the validity of salivary corticosterone (CORT) measurements in birds has not yet been reported. We aimed to measure salivary CORT in crows using a commercially available CORT enzyme-linked immunosorbent assay kit. An adrenocorticotropic hormone (ACTH) challenge experiment using synthetic cosyntropin, an ACTH analogue, was conducted to compare CORT level elevations between the serum and the saliva in a 10-60 min range. Both salivary and blood CORT was significantly elevated 10 min after injecting synthetic cosyntropin. The results supported the validation of salivary CORT as a surrogate for a blood CORT in crows.

Social encounters produce different autonomic response between dominants and subordinates in crows.

Recent studies of behavioural physiology on animals have suggested the crucial role of peripherally physiological signals in eliciting arousal and emotion. Heart rate (HR) is one of the useful and critical signals to measure autonomic regulation as a physiological basis for arousal and emotion in response to biologically significant stimuli such as social encounter with conspecific individuals. However, our understanding of peripherally physiological response such as HRs and autonomic activities under social contexts of non-human animals is still limited, particularly in birds. Here, we examined the autonomic activity of behaving crows exposed to a dominant and a subordinate conspecific by using non-invasive electrocardiogram recording. We found different patterns of autonomic responses dependent on the relative dominance position: dominant crows encountering subordinates showed the elevation of sympathetic activity, whereas subordinates encountering dominants showed decreased HR with elevated parasympathetic activity. This is the first study in birds to report different autonomic responses dependent on relative dominance positions during dyadic social encounters. The present study advances our understanding of the role of the peripheral autonomic system, as an interactive system with the brain, in eliciting emotion/arousal associated with socially challenging environments from an evolutionary perspective.

Measurement of urinary mesotocin in large-billed crows by enzyme-linked immunosorbent assay.

Mesotocin (MT) is an avian homologue of oxytocin (OT). Behavioral pharmacological studies in birds have suggested the involvement of MT in socially affiliative behavior. However, investigations of peripheral MT levels associated with social behavior are lacking because non-invasive methods to measure surrogate plasma MT have yet to be established. This study aimed to measure urinary MT in crows using a commercially available OT enzyme-linked immunosorbent assay kit. Urine samples were collected after intravenous injection of MT and centrifuged to separate urine and fecal components. We found that urinary MT was significantly elevated 15-30 min after MT injection. These results validate our method for the use of urine samples for the measurement of peripheral MT levels in crows.

Socio-ecological correlates of neophobia in corvids.

Behavioral responses to novelty, including fear and subsequent avoidance of novel stimuli, i.e., neophobia, determine how animals interact with their environment. Neophobia aids in navigating risk and impacts on adaptability and survival. There is variation within and between individuals and species; however, lack of large-scale, comparative studies critically limits investigation of the socio-ecological drivers of neophobia. In this study, we tested responses to novel objects and food (alongside familiar food) versus a baseline (familiar food alone) in 10 corvid species (241 subjects) across 10 labs worldwide. There were species differences in the latency to touch familiar food in the novel object and novel food conditions relative to the baseline. Four of seven socio-ecological factors influenced object neophobia: (1) use of urban habitat (versus not), (2) territorial pair versus family group sociality, (3) large versus small maximum flock size, and (4) moderate versus specialized caching (whereas range, hunting live animals, and genus did not), while only maximum flock size influenced food neophobia. We found that, overall, individuals were temporally and contextually repeatable (i.e., consistent) in their novelty responses in all conditions, indicating neophobia is a stable behavioral trait. With this study, we have established a network of corvid researchers, demonstrating potential for further collaboration to explore the evolution of cognition in corvids and other bird species. These novel findings enable us, for the first time in corvids, to identify the socio-ecological correlates of neophobia and grant insight into specific elements that drive higher neophobic responses in this avian family group. VIDEO ABSTRACT.

Scalable representation of time in the hippocampus.

Hippocampal "time cells" encode specific moments of temporally organized experiences that may support hippocampal functions for episodic memory. However, little is known about the reorganization of the temporal representation of time cells during changes in temporal structures of episodes. We investigated CA1 neuronal activity during temporal bisection tasks, in which the sets of time intervals to be discriminated were designed to be extended or contracted across the blocks of trials. Assemblies of neurons encoded elapsed time during the interval, and the representation was scaled when the set of interval times was varied. Theta phase precession and theta sequences of time cells were also scalable, and the fine temporal relationships were preserved between pairs in theta cycles. Moreover, theta sequences reflected the rats' decisions on the basis of their time estimation. These findings demonstrate that scalable features of time cells may support the capability of flexible temporal representation for memory formation.

Sex-specific effects of cooperative breeding and colonial nesting on prosociality in corvids.

The investigation of prosocial behavior is of particular interest from an evolutionary perspective. Comparisons of prosociality across non-human animal species have, however, so far largely focused on primates, and their interpretation is hampered by the diversity of paradigms and procedures used. Here, we present the first systematic comparison of prosocial behavior across multiple species in a taxonomic group outside the primate order, namely the bird family Corvidae. We measured prosociality in eight corvid species, which vary in the expression of cooperative breeding and colonial nesting. We show that cooperative breeding is positively associated with prosocial behavior across species. Also, colonial nesting is associated with a stronger propensity for prosocial behavior, but only in males. The combined results of our study strongly suggest that both cooperative breeding and colonial nesting, which may both rely on heightened social tolerance at the nest, are likely evolutionary pathways to prosocial behavior in corvids.

Control of bill-grasping aperture with varying food size in crows.

Grasping movement in primates is known to be a visually guided behavior and the aperture of hand opening is adjusted to the target size on the basis of visual information. The analogous behavior can be found in birds, called 'pecking', consisting of head-reaching and bill-grasping. Bill-grasping has been investigated mainly in pigeons and an aperture adjustment as seen in primates has been reported. This study focused on kinematics of pecking in crows, known to possess dexterous visuomotor skills, to examine whether crows adjust the grasping aperture to food diameter with a kinematic mechanism similar to that in pigeons. The pecking at a small piece of food was video recorded to analyze the grasping aperture. The results showed that the grasping aperture was proportional to food diameter. Kinematic analysis showed that the aperture adjustment was mediated by grasping velocity and grasping duration, which is consistent with the findings of previous research on pecking in pigeons. However, the relative contribution of grasping velocity was much higher than that of grasping duration. Our findings suggest the different sensorimotor mechanisms to control bill-grasping between the avian species with different foraging ecology.

Rapid adjustment of pecking trajectory to prism-induced visual shifts in crows as compared with pigeons.

Pecking in birds is analogous to reaching and grasping movements in primates. Earlier studies on visuomotor control in birds, which were conducted mostly in pigeons, suggested that avian pecking is controlled feedforwardly, and is out of the control of visual guidance during movement. However, recent studies using crows suggested a role of vision in pecking control during movement. To unveil what visuomotor mechanisms underlie the flexibility of pecking in crows, we examined whether pigeons and crows adjust their pecking to the visual distortion induced by prisms. Because prisms induce visual shifts of object positions, birds were required to adjust their movements. Pecking kinematics were examined before and after attaching prisms in front of the birds' eyes. Analysis of lateral deviation caused by the prisms showed that crows rapidly adjusted their pecking trajectories, but pigeons did so slowly. Angular displacement also increased in pigeons after attachment of the prism, but decreased in crows. These responses to prisms were consistent among individuals in pigeons but varied in crows, though the adjustment of pecking commonly succeeded in crows. These results suggest that pecking in pigeons predominantly involves feedforward control and that the movement is determined depending on the visual information available before the initiation of pecking. In contrast, the results from crows suggest that their pecking trajectories are corrected during the movement, supporting on-line visual control. Our findings provide the first evidence to suggest the on-line visual control of pecking in birds.

Avian brains: Insights from development, behaviors and evolution.

Birds are an extensively specialized animal group with unique anatomical, physiological and ecological characteristics. Sophisticated social behaviors and remarkable cognitive abilities are present in several avian lineages, driven by their enlarged brains and intricate neural networks. These unique traits could be a result of adaptive evolution under the wide range of environmental constraints; however, the intrinsic mechanisms of avian brain development and evolution remain unclear. Here, we introduce recent findings regarding developmental aspects of avian brain organization and neuronal networks for specific avian behaviors, which provide an insight into the link between the evolution of brain development and complex cognitive functions.

Flexible motor adjustment of pecking with an artificially extended bill in crows but not in pigeons.

The dextrous foraging skills of primates, including humans, are underpinned by flexible vision-guided control of the arms/hands and even tools as body-part extensions. This capacity involves a visuomotor conversion process that transfers the locations of the hands/arms and a target in retinal coordinates into body coordinates to generate a reaching/grasping movement and to correct online. Similar capacities have evolved in birds, such as tool use in corvids and finches, which represents the flexible motor control of extended body parts. However, the flexibility of avian head-reaching and bill-grasping with body-part extensions remains poorly understood. This study comparatively investigated the flexibility of pecking with an artificially extended bill in crows and pigeons. Pecking performance and kinematics were examined when the bill extension was attached, and after its removal. The bill extension deteriorated pecking in pigeons in both performance and kinematics over 10 days. After the bill removal, pigeons started bill-grasping earlier, indicating motor adaptation to the bill extension. Contrastingly, pecking in crows was deteriorated transiently with the bill extension, but was recovered by adjusting pecking at closer distances, suggesting a quick adjustment to the bill extension. These results indicate flexible visuomotor control to extended body parts in crows but not in pigeons.

Adaptive bill morphology for enhanced tool manipulation in New Caledonian crows.

Early increased sophistication of human tools is thought to be underpinned by adaptive morphology for efficient tool manipulation. Such adaptive specialisation is unknown in nonhuman primates but may have evolved in the New Caledonian crow, which has sophisticated tool manufacture. The straightness of its bill, for example, may be adaptive for enhanced visually-directed use of tools. Here, we examine in detail the shape and internal structure of the New Caledonian crow's bill using Principal Components Analysis and Computed Tomography within a comparative framework. We found that the bill has a combination of interrelated shape and structural features unique within Corvus, and possibly birds generally. The upper mandible is relatively deep and short with a straight cutting edge, and the lower mandible is strengthened and upturned. These novel combined attributes would be functional for (i) counteracting the unique loading patterns acting on the bill when manipulating tools, (ii) a strong precision grip to hold tools securely, and (iii) enhanced visually-guided tool use. Our findings indicate that the New Caledonian crow's innovative bill has been adapted for tool manipulation to at least some degree. Early increased sophistication of tools may require the co-evolution of morphology that provides improved manipulatory skills.

Sex-reversed correlation between stress levels and dominance rank in a captive non-breeder flock of crows.

Group living has both benefits and costs to individuals; benefits include efficient acquisition of resources, and costs include stress from social conflicts among group members. Such social challenges result in hierarchical dominance ranking among group members as a solution to avoid escalating conflict that causes different levels of basal stress between individuals at different ranks. Stress-associated glucocorticoid (corticosterone in rodents and birds; CORT) levels are known to correlate with dominance rank in diverse taxa and to covary with various social factors, such as sex and dominance maintenance styles. Although there is much evidence for sex differences in the basal levels of CORT in various species, the correlation of sex differences in basal CORT with dominance rank is poorly understood. We investigated the correlation between CORT metabolites (CM) in the droppings and social factors, including rank and sex, in a captive non-breeder group of crows. In this group, all the single males dominated all the single females, and dominance ranks were stable among single males but relatively unstable among single females. CM levels and rank were significantly correlated in a sex-reversed fashion: males at higher rank (i.e., more dominant) had higher CM, whereas females at higher rank exhibited lower CM. This is the first evidence of sex-reversed patterns of CM-rank correlation in birds. The results suggest that different mechanisms of stress-dominance relationships operate on the sexes in non-breeder crow aggregations; in males, stress is associated with the cost of aggressive displays, whereas females experience subordination stress due to males' overt aggression.

Involvement of vision in tool use in crow.

Birds are capable of dexterous sensory-motor activities such as tool use. Reaching is a crucial component of tool use and is a vision-guided behavior in primates, in which arm movement is monitored online in a stable visual frame. However, vision-guided reaching in primates is enabled by anatomical separation of the head and arm; neck reaching in birds accompanies head movement, which produces unstable vision because the eye necessarily moves with the bill. This anatomical difference raises the question whether tool use in birds involves visuomotor mechanisms that are distinct from those in primates. As the role of vision in avian tool use has been poorly understood, we investigated the role of vision in tool use in the large-billed crow (Corvus macrorhynchos), a nontool user in the wild. Crows were trained to manipulate an L-shaped hook to retrieve food that was otherwise out of reach. After training, an opaque panel was placed on the front window of the platform to block their vision, and the effects on tool use were tested with respect to performance and movement trajectory. Vision blocking caused similar deviation of tool movement trajectories for both near and far targets, as well as far target-specific deviation. This suggests the involvement of vision in tool use by crows, specifically in the premanipulation process for conversion of vision-body coordinates for motor planning and in the process of tool manipulation. This is the first behavioral evidence for the involvement of vision in avian tool use.

Crows cross-modally recognize group members but not non-group members.

Recognizing other individuals by integrating different sensory modalities is a crucial ability of social animals, including humans. Although cross-modal individual recognition has been demonstrated in mammals, the extent of its use by birds remains unknown. Herein, we report the first evidence of cross-modal recognition of group members by a highly social bird, the large-billed crow (Corvus macrorhynchos). A cross-modal expectancy violation paradigm was used to test whether crows were sensitive to identity congruence between visual presentation of a group member and the subsequent playback of a contact call. Crows looked more rapidly and for a longer duration when the visual and auditory stimuli were incongruent than when congruent. Moreover, these responses were not observed with non-group member stimuli. These results indicate that crows spontaneously associate visual and auditory information of group members but not of non-group members, which is a demonstration of cross-modal audiovisual recognition of group members in birds.

Gene expression profile in cerebrum in the filial imprinting of domestic chicks (Gallus gallus domesticus).

In newly hatched chicks, gene expression in the brain has previously been shown to be up-regulated following filial imprinting. By applying cDNA microarrays containing 13,007 expressed sequence tags, we examined the comprehensive gene expression profiling of the intermediate medial mesopallium in the chick cerebrum, which has been shown to play a key role in filial imprinting. We found 52 up-regulated genes and 6 down-regulated genes of at least 2.0-fold changes 3h after the training of filial imprinting, compared to the gene expression of the dark-reared chick brain. The up-regulated genes are known to be involved in a variety of pathways, including signal transduction, cytoskeletal organization, nuclear function, cell metabolism, RNA binding, endoplasmic reticulum or Golgi function, synaptic function, ion channel, and transporter. In contrast, fewer genes were down-regulated in the imprinting, coinciding with the previous data that the total RNA synthesis increased associated with filial imprinting. Our data suggests that the filial imprinting involves the modulation of multiple signaling pathways.

Up-regulation of microtubule-associated protein 2 accompanying the filial imprinting of domestic chicks (Gallus gallus domesticus).

Using cDNA microarrays, we have identified elsewhere the genes of microtubule-associated proteins as a group up-regulated in newly hatched chick brains after filial imprinting training. Here we show by in situ hybridization that the mRNA for the microtubule-associated protein 2 (MAP2) gene was enriched in the mesopallium and the hippocampus in the trained chick brain. The regionally specific enrichments of MAP2 mRNA were not observed in the brain of dark-reared or light-exposed chick as controls, implying an association between the degree of expression and the strength of the learned preference. In agreement with the gene expression, MAP2 protein was accumulated in the mesopallium of the trained chick brain, but not in the brains of the controls. The accumulation of MAP2 was found in the cytosol of neurons and co-localized with beta-tubulin, suggesting a change in microtubule assembly. Our results suggest a postnatal reorganization of cytoskeleton following filial imprinting.

Formation of linear dominance relationship in captive jungle crows (Corvus macrorhynchos): implications for individual recognition.

Jungle crows (Corvus macrorhynchos) flexibly change their social forms depending on their age, time of the day, and the season. In the daytime, paired adults behave territorially and unpaired subadults form small flocks of ten birds, whereas at night hundreds of birds roost together. In the breeding season, pairings remain in their nest all day. This fission-fusion raises questions about the underlying social structure and the cognitive capability of jungle crows. In this study, dyadic encounters were used to investigate dominance relationships (linear or non-linear) and the underlying mechanisms in captive jungle crows. Fourteen crows were tested in 455 encounters (i.e., 5 encounters per dyad), and a stable linear dominance relationship emerged. Sex and aggressiveness were determinants as individual characteristics for dominance formation. Males dominated females, and more aggressive individuals dominated less aggressive ones. Aggressive interactions in dyads occurred primarily during the first encounter and drastically declined during subsequent encounters without any signs of a confidence effect. These results suggest that, in captive jungle crow, a linear form of dominance is intrinsically determined by sex and aggressiveness and maintained extrinsically by memories of past outcomes associated with specific individuals, implying individual recognition.

Localized lesions of ventral striatum, but not arcopallium, enhanced impulsiveness in choices based on anticipated spatial proximity of food rewards in domestic chicks.

The effects of bilateral chemical lesions of the ventral striatum (nucleus accumbens and the surrounding areas in the medial striatum) and arcopallium (major descending area of the avian telencephalon) were examined in 1-2-weeks old domestic chicks. Using a Y-maze, we analyzed the lesion effects on the choices that subject chicks made in two tasks with identical economical consequences, i.e., a small-and-close food reward vs. a large-and-distant food reward. In task 1, red, yellow, and green beads were associated with a feeder placed at various distances from the chicks; chicks thus anticipated the spatial proximity of food by the bead's color, whereas the quantity of the food was fixed. In task 2, red and yellow flags on the feeders were associated with various amount of food; the chicks thus anticipated the quantity of food by the flag's color, whereas the proximity of the reward could be directly visually determined. In task 1, bilateral lesions of the ventral striatum (but not the arcopallium) enhanced the impulsiveness of the chicks' choices, suggesting that choices based on the anticipated proximity were selectively changed. In task 2, similar lesions of the ventral striatum did not change choices. In both experiments, motor functions of the chicks remained unchanged, suggesting that the lesions did not affect the foraging efficiency, i.e., objective values of food. Neural correlates of anticipated food rewards in the ventral striatum (but not those in the arcopallium) could allow chicks to invest appropriate amount of work-cost in approaching distant food resources.

Neural correlates of the proximity and quantity of anticipated food rewards in the ventral striatum of domestic chicks.

To identify the neuro-cognitive substrates of valuation and choice, we analysed the neural correlates of anticipated food rewards in the ventral striatum of freely behaving chicks. One-week-old chicks were trained in a color-discrimination task using four color cues (red, yellow, green and blue), each of which was associated with a different food reward. Choosing a red bead was immediately rewarded with a large amount of food, choosing a yellow bead resulted in an immediate-small food reward, and choosing a green bead resulted in a late-large food reward. We selected chicks that consistently chose a large and immediate food reward (red over yellow, and red over green), with the proximity of the food valued higher than the size of the food reward (yellow over green). Of the 47 neurons recorded from the ventral striatum of these chicks, 20 neurons selectively showed cue-period responses to cues associated with food rewards. Five of these 20 neurons responded differentially during the cue period according to the expected delay to reward, and were thus assumed to code for the proximity of the reward. Additionally, three other neurons responded to the quantity of the reward. Furthermore, in the post-operant delay period, many of these 20 neurons showed reward-related activities that were linked to the proximity or presence of the food reward. We therefore propose that impulsive choice and behavioral perseveration observed after lesions of the ventral striatum could be due to impaired anticipation of rewards in the cue and delay periods, respectively.