Both sheep and goats can solve inferential by exclusion tasks.

Despite the domestication of sheep and goats by humans for several millennia, we still lack comparative data on their cognitive capacity. Comparing the cognitive skills of farm animals can help understand the evolution of cognition. In this study, we compared the performances of sheep and goats in inference by exclusion tasks. We implemented two tasks, namely a cup task and a tube task, to identify whether success in solving the task could be attributed to either low-level mechanisms (avoiding the empty location strategy) or to deductive reasoning (if two possibilities A and B, but not A, then it must be B). In contrast to a previous study comparing goats and sheep in a cup task, we showed that both species solved the inferential condition with high success rates. In the tube task, performances could not be explained by alternative strategies such as avoiding the empty tube or preferring the bent tube. When applying a strict set of criteria concerning responses in all conditions and controlling for the potential effects of experience, we demonstrate that two individuals, a goat and a sheep, fulfil these criteria. This suggests that sheep and goats are able to make inferences based on deductive reasoning.

Unwilling or willing but unable: can horses interpret human actions as goal directed?

Social animals can gain important benefits by inferring the goals behind the behavior of others. However, this ability has only been investigated in a handful of species outside of primates. In this study, we tested for the first time whether domestic horses can interpret human actions as goal directed. We used the classical "unwilling versus unable" paradigm: an experimenter performed three similar actions that have the same outcome, but the goal of the experimenter differed. In the unwilling condition, the experimenter had no intention to give a piece of food to a horse and moved it out of reach when the horse tried to eat it. In the two unable conditions, the experimenter had the intention to give the food to the horse but was unable to do so, either because there was a physical barrier between them or because of the experimenter's clumsiness. The horses (n = 21) reacted differently in the three conditions: they showed more interest in the unable conditions, especially in the unable clumsy condition, than in the unwilling condition. These results are similar to results found in primates with the same paradigm and suggest that horses might have taken the experimenter's goal, or even intentions, into account to adapt their behavior. Hence, our study offers more insights into horse interspecific social cognition towards humans.

Horses feel emotions when they watch positive and negative horse-human interactions in a video and transpose what they saw to real life.

Animals can indirectly gather meaningful information about other individuals by eavesdropping on their third-party interactions. In particular, eavesdropping can be used to indirectly attribute a negative or positive valence to an individual and to adjust one's future behavior towards that individual. Few studies have focused on this ability in nonhuman animals, especially in nonprimate species. Here, we investigated this ability for the first time in domestic horses (Equus caballus) by projecting videos of positive and negative interactions between an unknown human experimenter (a "positive" experimenter or a "negative" experimenter) and an actor horse. The horses reacted emotionally while watching the videos, expressing behavioral (facial expressions and contact-seeking behavior) and physiological (heart rate) cues of positive emotions while watching the positive video and of negative emotions while watching the negative video. This result shows that the horses perceived the content of the videos and suggests an emotional contagion between the actor horse and the subjects. After the videos were projected, the horses took a choice test, facing the positive and negative experimenters in real life. The horses successfully used the interactions seen in the videos to discriminate between the experimenters. They touched the negative experimenter significantly more, which seems counterintuitive but can be interpreted as an appeasement attempt, based on the existing literature. This result suggests that horses can indirectly attribute a valence to a human experimenter by eavesdropping on a previous third-party interaction with a conspecific.

All bonds are not alike: A psychoendocrine evaluation of infant attachment.

Characteristics of attachment were assessed in peer- and object-reared lambs, and compared to mothered subjects by taking into consideration distress, proximity seeking, and exploration during two separation-reunion tests in both the familiar and a novel environment. Plasma cortisol and oxytocin were assayed as physiological indicators of stress and being comforted during the separation-reunion test. Rewarding properties of the familiar figures were also determined in a conditioned place preference-like paradigm. Between-group analysis revealed the existence of secure attachment with the mother, alteration of secure attachment with the peer and weaker attachment with the object. Weaker attachment was expressed by a lack of distress during separation in the home pen and no preference for the place conditioned with the familiar object. Elevated basal plasma oxytocin levels, but not cortisol, observed in maternally deprived lambs were more likely linked to the absence of a maternal figure rather than social comfort during reunion.

Physiological changes in the peri-partum period and colostral IgG transfer in prolific D'man sheep: effects of parity and litter size.

The aim of this work was to assess maternal and neonatal changes in plasma proteins, glucose and cortisol and to quantify the colostral immunoglobulin G (IgG) transfer in the peri-partum period in D'man sheep, a prolific breed, taking into account the parity of the ewe. The concentrations of proteins and glucose were high in the ewes on day 7 and at lambing before decreasing. Likewise, cortisol plasma concentration was maximal during the 6 h following lambing and dropped at 12 h. Protein and glucose concentrations were low in lambs at 1 h of birth after which they increased. By contrast, cortisol level was the highest during the first 12 h of birth and then decreased. The colostral IgG level was high at lambing and dropped by over 87 % from 1 to 48 h post-partum. In the newborn, the plasma IgG concentration was lowest at birth and increased rapidly during the first 24 h of birth. Parity influenced maternal physiology with multiparous ewes having the lowest concentrations of proteins, glucose, IgG and cortisol, but the highest colostrum IgG level. Accordingly, lambs born from primiparous ewes had lower protein, glucose and plasma IgG concentrations than lambs born from multiparous ewes. The main outcome of this study was that lambs born from primiparous ewes are characterized by the lowest physiological indices and this may influence their survival chance.

Effects of prenatal stress and emotional reactivity of the mother on emotional and cognitive abilities in lambs.

Consequences of prenatal stress on emotional reactivity and cognitive abilities in offspring are under-documented in precocial mammals. Here, we investigated to what extent emotional reactivity, judgment bias and spatial learning abilities of lambs are affected by chronic stress during late pregnancy and by their dams' emotional reactivity. The 20 highest-responsive (HR) and 20 lowest-responsive (LR) ewes from a population of 120 Romane ewes were selected according to their pre-mating reactivity to social isolation in a new environment. Over the final third of pregnancy, 10 HR ewes and 10 LR ewes were exposed daily to various unpredictable aversive events such as restraint, mixing groups and transport while the other 20 selected ewes were not. In a human and an object test, prenatally-stressed lambs were more fearful than control lambs, but the prenatal stress effect was moderated by the reactivity of the mothers: prenatally-stressed lambs from ewes with high emotional reactivity were more affected. Prenatally-stressed lambs did not perform as well as control lambs in a maze test and showed pessimistic-like judgment in a cognitive bias test. Prenatally-stressed lambs were thus characterized by a negative affective state with increased fear reactions and impaired cognitive evaluation. The development of negative moods could have long-lasting consequences on the coping strategies of the lambs in response to their rearing conditions.

Mild effects of gestational stress and social reactivity on the onset of mother-young interactions and bonding in sheep.

Consequences of prenatal stress on mother-young relationships are well-documented in altricial mammals but less so in precocial mammals. In this study, we investigated the effects of unpredictable aversive events on maternal behavior and mutual mother-young recognition in pregnant ewes while accounting for modulatory effects of ewe reactivity. From a population of 120 Romane-breed ewes, we selected 20 high-responsive (HR) and 20 low-responsive (LR) ewes according to pre-mating reactivity assessed in isolation tests. Over the final third of pregnancy, 10 HR ewes and 10 LR ewes were exposed daily to various aversive events such as social isolation, mixing and transport (stressed ewes), while the other 20 ewes were not exposed to aversive events (control ewes). Although the treatment induced chronic stress, physiologically confirmed by an increase in salivary cortisol following transport and sham shearing, maternal behavior of stressed ewes observed during the first 30 min postpartum and in the selectivity test 1 h 30 min later did not differ from controls. However, in a maternal motivation test performed 48 h postpartum, stressed ewes vocalized less than controls when separated from their lambs, and walked less readily past an unknown object to reach their lambs. Lambs of stressed ewes spent more time near their dam in a preference test performed 15 h after birth compared to control-ewe lambs. HR ewes spent more time grooming their lambs than LR ewes. We posit that domestication could have selected animals displaying robust expression of maternal behavior related to social reactivity and producing offspring that are better adapted to challenging situations.

Captive Blue-and-yellow macaws (Ara ararauna) show facial indicators of positive affect when reunited with their caregiver.

In mammals, human-animal bonding is recognized as a source of positive affect for companion or farm animals. Because this remains unexplored in birds, we investigated captive parrots' perspective of the human-animal relationship. We used a classical separation-reunion paradigm and predicted that variations in parrots' facial displays and behaviours would indicate their appraisal of the relationship. The test was divided into three phases of two minutes each: the bird was placed in an unfamiliar environment with a familiar caregiver (union), then the bird was left alone (separation) and finally, the caregiver returned (reunion). The test was repeated 10 times for each bird and video recorded in order to analyze their behaviour. The data show significantly higher crown and nape feather heights, higher redness of the skin and higher frequency of contact-seeking behaviours during the union and reunion phases than during the separation phase during which they expressed long distance contact calls. We observed the expression of eye pinning during the union and reunion phases in one out of five macaws. We argue that variation in facial displays provides indicators of parrot's positive appraisal of the caretaker presence. Our results broaden the scope for further studies on parrots' expression of their subjective feelings.

Maternal deprivation and milk replacement affect the integrity of gray and white matter in the developing lamb brain.

The psychoendocrine evaluation of lamb development has demonstrated that maternal deprivation and milk replacement alters health, behavior, and endocrine profiles. While lambs are able to discriminate familiar and non-familiar conspecifics (mother or lamb), only lambs reared with their mother develop such clear social discrimination or preference. Lambs reared without mother display no preference for a specific lamb from its own group. Differences in exploratory and emotional behaviors between mother-reared and mother-deprived lambs have also been reported. As these behavioural abilities are supported by the brain, we hypothesize that rearing with maternal deprivation and milk replacement leads to altered brain development and maturation. To test this hypothesis, we examined brain morphometric and microstructural variables extracted from in vivo T1-weighted and diffusion-weighted magnetic resonance images acquired longitudinally (1 week, 1.5 months, and 4.5 months of age) in mother-reared and mother-deprived lambs. From the morphometric variables the caudate nuclei volume was found to be smaller for mother-deprived than for mother-reared lambs. T1-weighted signal intensity and radial diffusivity were higher for mother-deprived than for mother-reared lambs in both the white and gray matters. The fractional anisotropy of the white matter was lower for mother-deprived than for mother-reared lambs. Based on these morphometric and microstructural characteristics we conclude that maternal deprivation delays and affects lamb brain growth and maturation.

Horses Solve Visible but Not Invisible Displacement Tasks in an Object Permanence Paradigm.

A key question in the field of animal cognition is how animals comprehend their physical world. Object permanence is one of the fundamental features of physical cognition. It is the ability to reason about hidden objects and to mentally reconstruct their invisible displacements. This cognitive skill has been studied in a wide range of species but never directly in the horse (Equus caballus). In this study, we therefore assessed the understanding of visible and invisible displacements in adult Welsh mares in two complementary experiments, using different horses. In experiment 1, visible displacement was investigated using two tasks adapted from the Uzgiris and Hunt scale 1. Invisible displacement was assessed using a transposition task, in which food was first hidden in one of two containers and the location of the containers was then switched. In experiment 2, we further investigated horses' understanding of visible and invisible displacements using an easier procedure designed to avoid potentially confounding factors. In both experiments, horses successfully completed the tasks involving visible displacement with two or three possible hiding places. However, in both experiments, horses failed the transposition tasks, suggesting that they may not be able to track the displacement of an object that is not directly perceived (i.e., invisible displacement). These results bring new insights into object permanence in horses and how they represent their physical world.

Neonatal Suckling, Oxytocin, and Early Infant Attachment to the Mother.

The neuropeptide oxytocin (OT) promotes maternal care and social affiliation in adults but its importance in infant attachment still remains unknown. True animal models of infant attachment are extremely rare, and the sheep (in complement to non-human primates) is one of the few that provides the opportunity to investigate its neuroendocrinological basis. In the lamb, access to the udder has strong rewarding properties for the establishment of a preferential relationship with the mother. Therefore, the present study explored the possible involvement of OT through its release during close social contacts with the mother. The first experiment revealed that lambs having free access to the udder from birth onward developed, by 12 h of age, a clear preference for their mothers over another maternal ewe. Delaying access to the udder for six, four or even only 2 h starting at birth, by covering the ewe's udder, resulted in the lack of such a preference without affecting general activity. These effects persisted in most cases at 24 h but by 72 h of age a bond with the mother was clearly expressed. Experiment two showed that social interactions with the mother were followed by a release of OT in the plasma when lambs had the possibility to suckle. Non-nutritive interactions were without effects. Preliminary data on two subjects suggested that OT might also increase in the cerebrospinal fluid after suckling. Finally, in the third experiment, oral administration of a non-peptide OT receptor antagonist (L-368-899, Merck) over the first 4 h after birth led to decreased exploration of the mother's body compared to lambs receiving saline, and impaired the expression of a preference for the mother at 24 h. The effects were no longer observed at 48 h. Our findings demonstrate that both delayed access to the mother's udder and OT receptor antagonist alter the onset of mother preference in newborn lambs. This suggests that central OT facilitates the development of filial attachment through its release during suckling.

Behavioural and neurobiological effects of colostrum ingestion in the newborn lamb associated with filial bonding.

In sheep, the onset of filial bonding relies on early intake of colostrum. The aim of our work was to describe in the newborn lamb housed with its mother the immediate post-ingestive effects of colostrum intake, in terms of behaviour and brain activity. In Experiment 1, lambs received five nasogastric infusions of colostrum, or saline, or sham intubations during the first 6 h after birth. Mother-young interactions were recorded before and after the first, third and fifth infusions. The activity of the dam and of the young, which diminished over time in all groups, was temporarily increased in both partners just after each intubation procedure. The number of high-pitched bleats was significantly lower in lambs that received colostrum than in the sham group, suggesting soothing or satiating properties of colostrum. In Experiment 2, newborn lambs received a single nasogastric infusion of colostrum or saline 4.5 h after birth, or were sham intubated. Neuronal activation was investigated 1.5 h later for maximum c-Fos activity. Infusion of colostrum and saline induced different patterns of c-Fos-like immunoreactivity in the paraventricular and supraoptic nuclei of the hypothalamus as compared with the sham group. A specific oxytocinergic/vasopressinergic (OT/VSP) cell population in the paraventricular nucleus was activated following colostrum and saline infusion, but not sham intubation. Only colostrum induced the activation of the cortical amygdala and insular cortex, two structures involved in learning, associative processes, reward and emotion. We hypothesize that filial bonding may be triggered through colostrum-rewarded learning/calming processes and that the OT/VSP system may play a role.

Horses Categorize Human Emotions Cross-Modally Based on Facial Expression and Non-Verbal Vocalizations.

Over the last few years, an increasing number of studies have aimed to gain more insight into the field of animal emotions. In particular, it is of interest to determine whether animals can cross-modally categorize the emotions of others. For domestic animals that share a close relationship with humans, we might wonder whether this cross-modal recognition of emotions extends to humans, as well. In this study, we tested whether horses could recognize human emotions and attribute the emotional valence of visual (facial expression) and vocal (non-verbal vocalization) stimuli to the same perceptual category. Two animated pictures of different facial expressions (anger and joy) were simultaneously presented to the horses, while a speaker played an emotional human non-verbal vocalization matching one of the two facial expressions. Horses looked at the picture that was incongruent with the vocalization more, probably because they were intrigued by the paradoxical combination. Moreover, horses reacted in accordance with the valence of the vocalization, both behaviorally and physiologically (heart rate). These results show that horses can cross-modally recognize human emotions and react emotionally to the emotional states of humans, assessed by non-verbal vocalizations.

Olfactory preference for own mother and litter in 1-day-old rabbits and its impairment by thermotaxis.

We investigated the ability of rabbit pups to display preferences towards various elements of their postnatal environment during the stage of confinement in the nest. Subjects were submitted to a two-choice test during the first week after birth to assess if they could detect and discriminate between does, litters of pups, or nesting materials of the same developmental stage. On D1 and D7, pups were attracted to any lactating doe, litter, or nest when compared to an empty compartment. When two stimuli were opposed, pups preferred their own nest to an alien one on D1 and D7 but not their mother nor their siblings when opposed to alien does or pups. However, additional tests indicated that this lack of preference for kin conspecifics resulted from a predominant attraction to thermal cues over individual odors. Indeed, pups were strongly attracted to a warm compartment (37 degrees C) than to a cooler one (20 degrees C) and once thermal cues were controlled for in the testing situation, the pups were specifically attracted to odors of their own mother's hair and of their siblings. No preference was observed towards the mother's uterine secretions. In conclusion, pups can recognize olfactory cues emanating from their mother and their siblings the day after birth. The preference for nesting materials would reflect in major part the combined attraction to maternal and sibling odors present in the nest.

Facial expression and oxytocin as possible markers of positive emotions in horses.

Behavioural and physiological markers of discrete positive emotions remain little investigated in animals. To characterise new markers in horses, we used tactile stimulations to induce emotional situation of contrasting valence. In the Gentle grooming group (G, N = 13) horses were gently groomed during 11 sessions on the body areas they appreciated the most. Horses in the Standard grooming group (S, N = 14) were groomed using a fixed procedure, reported to induce avoidance reactions in some horses. At session 11, G horses expressed significantly more contact-seeking behaviours than S horses, which expressed significantly more avoidance behaviours. This result suggests positive emotions in G horses and negative emotions in S horses. Blood cortisol, oxytocin, heart rate and heart rate variability never differed between before and after the grooming session. However, after the 11 sessions, basal oxytocin levels were lower in the G than in the S group. This difference was unexpected, but supports studies showing that a low level of basal oxytocin could be a marker of better well-being. Analyses of facial expressions during grooming revealed significant differences between groups. These expressions appear to be more sensitive than behavioural indicators because they alone enabled differentiating emotions according to the group when horses were re-exposed to neutral grooming one year after the treatment.

Are there facial indicators of positive emotions in birds? A first exploration in Japanese quail.

The positive aspect of emotions, like pleasure, remains overlooked in birds. Our aim was to contribute to the exploration of facial indicators of positive emotions. To observe contrasting emotional expressions, we used two lines of Japanese quail divergently selected on their inherent fearfulness: a fearful line (long tonic immobility duration: LTI) and a less fearful line (short tonic immobility duration: STI). To induce positive emotions, we gave individual quail the opportunity to perform a rewarding behaviour, dustbathing, in an unfamiliar cage. More STI than LTI quail expressed dustbathing and latencies to dustbathe were significantly shorter in STI than LTI quail. This result indicated that the lines of quail differed in their fearfulness of the situation. We observed crown feather height, throat feather angle and pupil surface before (control) and during dustbathing. We found significant increases in crown feather height, pupil area and angle of throat feathers between the control and the dustbathing phases in STI quail, and pupil area correlated positively with crown feather height. In LTI quail, the angle of throat feathers increased during dustbathing, but the other parameters did not differ. We argue that variation in crown feather height and pupil area may provide indications of positive emotions in Japanese quail.

Behavioral evidence of heterospecific bonding between the lamb and the human caregiver and mapping of associated brain network.

While behavioral mechanisms of bonding between young mammals and humans have been explored, brain structures involved in the establishment of such processes are still unknown. The aim of the study was to identify brain regions activated by the presence of the caregiver. Since human positive interaction plays an important role in the bonding process, activation of specific brain structures by stroking was also examined. Twenty-four female lambs reared in groups of three were fed and stroked daily by a female caregiver between birth and 5-7 weeks of age. At 4 weeks, an isolation-reunion-separation test and a choice test revealed that lambs developed a strong bond with their caregiver. At 5-7 weeks of age, lambs were socially isolated for 90min. They either remained isolated or met their caregiver who stroked them, or not, at regular intervals over a 90-min period. Neuronal activation was investigated at the end of the period for maximum c-Fos expression. Reunion with the caregiver appeased similarly the lambs whether stroking was provided or not. Stroking did not activate a specific brain network compared to no stroking. In both cases, brain regions associated with olfactory, visual and tactile cue processing were activated in the presence of the caregiver, suggesting a multisensory process involved. In addition, activation of the oxytocinergic system in the hypothalamic paraventricular nucleus induced by the presence of the caregiver suggests similar neuroendocrine mechanisms involved in inter-conspecific and animal-human bonding.

Do lambs perceive regular human stroking as pleasant? Behavior and heart rate variability analyses.

Stroking by humans is beneficial to the human-animal relationship and improves welfare in many species that express intraspecific allogrooming, but very few studies have looked at species like sheep that do not express such contact except around parturition. This study investigated the way lambs perceive regular human tactile contact using behavioral and physiological responses. Twenty-four lambs were reared and bucket-fed in groups of four. All were stroked daily by their familiar caregiver. At 8 weeks of age, the lambs were individually tested in their home pen but in a 1×1m open-barred pen after a 15 h period of habituation to physical separation from peers while remaining in visual and auditory contact. Half of the lambs received stroking by their caregiver for 8 min and half were exposed to their caregiver's immobile presence. Heart rate and heart rate variability were recorded and analyzed by 2-min slots over the same interval based on three measures: mean heart rate value (HR), root mean square of successive differences (RMSSD) and standard deviation of all intervals measured between consecutive sinus beats (SDNN). Behavioral responses (ear postures of the lamb and time spent in contact with the familiar caregiver, on the knees of the familiar caregiver, and moving) were recorded throughout the test. Lamb HR decreased continuously while in the presence of their caregiver. Lambs being stroked showed slower HR and higher RMSSD which reflected positive emotional states compared to lambs left unstroked. All behavioral variables were highly correlated with the main component axis of the PCA analyses: the more the animals stayed in contact with their caregiver, the less they moved and the more their ears were hanging. This first component clearly differentiates lambs being stroked or not. Behavioral and physiological observations support the hypothesis that gentle physical contact with the caregiver is perceived positively by lambs.

Differential c-Fos expression in the newborn lamb nucleus tractus solitarius and area postrema following ingestion of colostrum or saline.

Visceral stimuli and the gut-brain axis play a crucial role in the control of ingestion even in the neonate. The aim of this study was to assess the neuronal activation in the nucleus tractus solitarius (NTS) and the area postrema (AP) following nutritional and non-nutritional stimulations. Lambs received a single gastric infusion of colostrum or saline at 5% birth weight or were sham infused. Infusion of either liquid led to c-Fos-like immunoreactivity (c-FLI) in the NTS and AP. Differences were observed along the sections of the NTS rostro-caudal axis according to the nature of the stimulation, suggesting a specificity of certain afferents and/or NTS areas for nutritional or non-nutritional signals. In the AP, the neuronal activation induced by colostrum was much higher than that induced by saline. A higher number of TH-immunoreactive cells were activated following colostrum infusion, suggesting a specific involvement of the catecholaminergic pathway in the treatment of meal-related stimuli. In spite of functional convergence, the two medullary structures observed responded differently according to the stimulation, indicating a complementary role in the integration of visceral signals.

Human-lamb bonding: oxytocin, cortisol and behavioural responses of lambs to human contacts and social separation.

Friendly interactions between humans and animals such as gentling or petting have been shown to have positive behavioural and physiological consequences in many species. In primates, rodents and dogs, oxytocin has been associated with tactile contact and anti-stress effects that may influence bonding and responses to stress situations. However the activation of the oxytocinergic system in other human-animal interactions such as with herbivores, had not yet been studied. Sixteen female lambs were reared by artificial feeding reinforced with 3× 30 s daily stroking sessions. At 6 weeks of age, the test consisted in measuring first plasma oxytocin and cortisol responses in lambs during a first 6-min phase in the home pen where the familiar caregiver gently stroked the lamb, and then physiological and behavioural responses in a test pen during a 20-min - phase of social isolation followed by a 20-min - phase of reunion with its familiar caregiver. The lambs expressed behavioural agitation during the whole period of isolation. A strong affiliative response towards the human and a sustained reduction of the agitation behaviour were observed during reunion. Lambs' behaviours when isolated and when in contact with the human were correlated suggesting a response to social separation from the familiar caregiver more than to social isolation from congeners. No significant changes in cortisol levels were observed during the test. Oxytocin levels did not vary during human contact, but increased when the familiar caregiver left the lamb alone in the test pen. In conclusion, lambs displayed affiliative responses towards their caregiver, and the lack of cortisol response during isolation while oxytocin was released suggest an anti-stress effect of oxytocin.