Selected Acoustic Frequencies Have a Positive Impact on Behavioural and Physiological Welfare Indicators in Thoroughbred Racehorses.

(1) Background: Since antiquity, it is considered that sounds influence human emotional states and health. Acoustic enrichment has also been proposed for domestic animals. However, in both humans and animals, effects vary according to the type of sound. Human studies suggest that frequencies, more than melodies, play a key role. Low and high frequencies, music tuning frequency and even EEG slow waves used for 'neurofeedback' produce effects. (2) Methods: We tested the possible impact of such pure frequencies on racehorses' behavior and physiology. A commercial non-audible acoustic stimulus, composed of an array of the above-mentioned frequencies, was broadcasted twice daily and for three weeks to 12 thoroughbred horses in their home stall. (3) Results: The results show a decrease in stereotypic behaviors and other indicators such as yawning or vacuum chewing, an increase in the time spent in recumbent resting and foraging, and better hematological measures during and after the playback phase for 4 of the 10 physiological parameters measured. (4) Conclusions: These results open new lines of research on possible ways of alleviating the stress related to housing and training conditions in racehorses and of improving physical recovery.

Horses' attentional characteristics differ according to the type of work.

Attention is a central process of cognition and influences the execution of daily tasks. In humans, different types of work require different attentional skills and sport performance is associated with the ability to attention shift. Attention towards humans varies in dogs used for different types of work. Whether this variation is due to the recruitment of individuals suitable for specific types of work, or to the characteristics of the work, remains unclear. In the present study, we hypothesized that domestic horses (Equus caballus) trained for different types of work would also demonstrate different attentional characteristics but we also explored other possible factors of influence such as age, sex and breed. We exposed more than sixty horses, working in 4 different disciplines, and living in two types of housing conditions, to a visual attention test (VAT) performed in the home environment. Individual attentional characteristics in the test were not significantly influenced by age, sex, breed or conditions of life but were strongly related to the type of work. Riding school horses showed longer sequences and less fragmented attention than all other horses, including sport horses living in the same conditions. Interestingly, sport performance was correlated with attention fragmentation during the test in eventing horses, which may need more attention shifting during the competitions. Working conditions may influence attention characteristics indirectly through welfare, or directly through selection and training. Our study opens new lines of thought on the determinants of animal cognition and its plasticity and constitutes a further step towards understanding the interrelationship between working conditions and cognition.

What Do We Know about Pruritus in Very Young Infants? A Literature Review.

In infants, pruritus is frequently considered as absent because they do not scratch themselves. Because pruritus could induce severe adverse effects in this vulnerable population, we aimed to review existing evidence on the ability of young infants to experience itch and on how to assess itch-related discomfort in this population. A literature review was performed (Pubmed, Google Scholar). Neurological itch pathways are well described. Skin development starts early during gestation. At 34 weeks of gestation, skin is almost complete while skin adaptations occur after birth. Newborn skin is neurologically functional, including the ability for young infants to feel pain. Similarities and interactions between pain and pruritus support the hypothesis that infants could feel pruritus. However, the existence of pruritus in infants has never been evidenced. Many itchy conditions can affect them, suggesting non-negligible prevalence of infant pruritus among which atopic dermatitis (AD) is the most studied disease. Studies reported a negative impact of AD on children and their families. There is no existing validated method to assess pruritus in infants, although they may feel pruritus and chronic pruritus can lead to serious adverse effects. To appropriately diagnose pruritus appears of great interest among young infants. Development of a method is required to this aim.

Detecting Welfare in a Non-Verbal Species: Social/Cultural Biases and Difficulties in Horse Welfare Assessment.

Horses were domesticated for more than 5000 years and have been one of the most emblematic species living alongside humans. This long-shared history would suggest that horses are well known and well understood, but scientific data raise many concerns about the welfare state of most domestic horses suggesting that many aspects have been largely misunderstood. In the present review, we will examine some of the possible human factors that may explain the huge prevalence of welfare problems, despite horses being of special importance to humans. First of all, as horses are non-verbal, current management practices rely upon what one thinks is good for them, which opens the way to subjective interpretations and projections, based on one's own subjective experience but probably still more on cultural/social norms and influences, traditions and beliefs. The lack of recognition, identification, or even the misinterpretation of signals are other potential reasons for welfare issues. Lastly, the over-exposure to animals with expressions of compromised welfare may lead to lower sensitivity of owners/professionals. That is why we lastly suggest that instead of simply providing information on what to do, we should promote validated visible indicators that leave less room for personal interpretation.

EEG individual power profiles correlate with tension along spine in horses.

Assessing chronic pain is a challenge given its subjective dimension. In humans, resting state electroencephalography (EEG) is a promising tool although the results of various studies are contradictory. Spontaneous chronic pain is understudied in animals but could be of the highest interest for a comparative study. Riding horses show a very high prevalence of back disorders thought to be associated with chronic pain. Moreover, horses with known back problems show cognitive alterations, such as a lower attentional engagement. Therefore, we hypothesized that the individual EEG power profiles resting state (i.e. quiet standing) of different horses could reflect the state of their back, that we measured using static sEMG, a tool first promoted to assess lower back pain in human patients. Results show that 1) EEG profiles are highly stable at the intra-individual level, 2) horses with elevated back tension showed resting state EEG profiles characterized by more fast (beta and gamma) and less slow (theta and alpha) waves. The proportion of theta waves was particularly negatively correlated with muscular tension along the spine. Moreover, elevated back tension was positively correlated with the frequency of stereotypic behaviours (an "addictive- like" repetitive behavior) performed by the horses in their stall. Resting state quantitative EEG appears therefore as a very promising tool that may allow to assess individual subjective chronic pain experience, beyond more objective measures of tension. These results open new lines of research for a multi-species comparative approach and might reveal very important in the context of animal welfare.

The vocal repertoire of preterm infants: Characteristics and possible applications.

We investigated infants' capacities to express themselves orally at very early developmental stages. Most reports focus on crying when in pain or hungry. We evaluated young preterm infants' spontaneous vocal production in non-painful contexts. We identified a vocal repertoire composed of nine types of vocalisations. High-pitched sounds were associated with relaxed postures, implying a positive valence, whereas long low-pitched vocalisations, associated more with grimaces and muscle tensions, appeared to have a more negative valence. Infants' vocalisations were useful indicators of their internal state in two situations (when exposed to clothing constraints and environmental noises).

A novel, short and easy-to-perform method to evaluate newborns' social olfactory preferences.

Humans' early olfactory perception has been studied mainly within the framework of mother-offspring interactions and only a few studies have focused on newborns' abilities to discriminate body odors per se. The aim of this study was to develop a method to evaluate olfactory social preferences of infants at term-equivalent age. Twenty dyads of infants (10 born preterm and 10 born at term) at term-equivalent age and their mothers were included. We analyzed the behavioral reactions of infants to their mother's upper-chest odor (that bears social, non-food related information). The two impregnated gauzes and a control gauze were presented to the infants for 10 s each, in a random order. We compared two durations of gauze impregnation: 30 min and 12 h. This study reveals that mothers' upper chest emits sufficient olfactory information to induce reactions in infants born full-term or born preterm and that a short impregnation is preferable to evaluate their perception of body odors, notably for those born preterm.

Higher tactile sensitivity in preterm infants at term-equivalent age: A pilot study.

Despite a growing body of research on perinatal sensory abilities, data on the extent of tactile sensitivity and more particularly passive touch (i.e. sensitivity to a stimulation imposed on the skin) are relatively limited, and the development and processing of tactile function are still thus little known. This question is particularly of high importance for infants with atypical early development such as those born prematurely who are exposed to many sensory (including tactile) stimulations (being in a hospital setting) during a critical period of brain development and those born at early term whose birth occurs at the precise time of cortical reorganization, in particular in the sensory areas. Some parents and health-care providers have for instance reported that children born prematurely exhibit atypical (e.g. higher) sensitivity to "benign" tactile stimuli. In the present study, we hypothesized that preterm and early-term infants may show altered tactile sensitivity. We compared the behavioral responses around term-equivalent age of infants born either pre-term, early-term or at term to the application of a light (0.008 grams) mechanical stimulus. We found that almost all preterm infants perceive this tactile stimulus, contrarily to the two other groups of infants. This extreme tactile sensitivity may be due to experiential, maturational or more likely both processes. We also compared the tactile sensitivity of these infants to that of adults. We found that adults were irresponsive to the light mechanical stimulus. This finding opens not only new insights in understanding development of tactile processing, but also new lines of thought about the particular sensory world of premature and early-term infants and hence about the potential impact of early care practices.

Domestic Foal Weaning: Need for Re-Thinking Breeding Practices?

Artificial weaning is a standard practice known to be one of the most stressful events in a domestic foal's life. Research has mainly focused on ways to alleviate weaning stress. However, there is still a need for more detailed research on what should constitute best practices with respect to animal welfare. The aim of this review is to address this issue by examining the natural weaning process. We first provide an overview of the scientific literature on the natural temporal dynamics of the dam-offspring bond in horses: it is to be noted that the natural process of weaning is little documented, individual variations have been poorly investigated and immediate effects of weaning on the mare-foal relationship remain unexplored. To partly address these gaps, we performed a study around the weaning period on 16 mare-foal pairs kept with minimal human interference. Most foals were weaned spontaneously when 9-10 months old, with individual variations mainly due to the conception rate of mares. Natural weaning induced no stress response in either partner and was performed without clear signs of rejection by the dams either just before or after. We lastly open up the discussion on the need for rethinking weaning practices under domestic conditions.

On-Farm Welfare Assessment of Horses: The Risks of Putting the Cart before the Horse.

Although the question of animal welfare has been an important source of concern in the scientific community for several decades, many aspects are still under debate. On-farm assessments have to be rapid, acceptable to farmers and safe for both the assessors and animals. They are thus very demanding, with multiple decisions to make, such as the choice of appropriate indicators, sampling methods and scoring. Research has moved from resource-based to animal-based criteria, which reflects the subjective welfare state of an animal rather than relying upon external indices. In the present review, we describe two major (i.e., the most frequently/recently tested or disseminated) protocols: one in low-/middle-income countries, and the other in high-income countries, for on-farm assessments of horses, using animal-based resources; we evaluate their strengths and limitations, and then we compare their results with those obtained by various other studies. We propose lines of improvement, particularly in view of public dissemination, and offer suggestions for further refinement or new protocols. We emphasize the high risks of putting the cart before the horse, i.e., proposing protocols that rely upon indicators and sampling methods that need to be refined, as this could lead to under-evaluation (or less likely over-evaluation) of current welfare problems. Because welfare is a subjective experience, the true representation of an individual's actual welfare status has to be evaluated by using objective assessment tools (that are validated and have a scientific basis) used by well-trained observers.

Changes in Saliva Analytes Correlate with Horses' Behavioural Reactions to An Acute Stressor: A Pilot Study.

Acute stress induces an array of behavioural reactions in horses that vary between individuals. Attempts to relate behavioural patterns and physiological responses have not always given clear-cut results. Here, we measured the changes in a panel of salivary components: salivary alpha-amylase (sAA), lipase, total esterase (TEA), butyrylcholinesterase (BChE), adenosine deaminase (ADA), and cortisol, and their potential link with horses' behaviours after acute stress. Saliva samples were collected in nine riding horses subjected to a test consisting of opening an umbrella. Saliva sampling was obtained at a basal time point in the stall (T1), in the test indoor arena (T2), at a time of stress (T3), and 30 min (T4) and 60 min (T5) later. The horses' behaviour was recorded at T3 for 1 minute. sAA, lipase, TEA, and BChE showed significant changes along time, increasing at T3 for BChE, and decreasing at T4 for sAA and BChE. Butyrylcholinesterase appeared to be the most reliable predictor of behavioural responses, as it correlated with the index of emotionality, of laterality, and the occurrence of alarm signals, while sAA decreased when horses expressed quieter behaviours. These first results bring promising lines for novel, more precise physiological markers of acute stress in horses that can bridge the gap between behaviour and physiology.

Snort acoustic structure codes for positive emotions in horses.

While the vocal coding of human and animal internal states has been widely studied, the possible acoustic expression of "positive" emotions remains poorly known. Recent studies suggest that snorts (non-vocal sounds produced by the air expiration through the nostrils) appear to be reliable indicators of positive internal states in several ungulate species. Here, we hypothesised in horses that the acoustic structure of the snort could vary with the subjects' current emotional state. Indeed, a preliminary sound analysis of snorts let us suggest structure variations related to the presence of pulsations. We recorded snorts from 20 horses living in a riding center. Auditory playbacks run with 20 humans first confirmed the existence of two snort subtypes, i.e. one pulsed and one non-pulsed. Observations were then conducted to compare the distribution of these two subtypes according to the location (stall/pasture) of the signaller as a contextual determinant of its internal state and to its ears' position as a reflection of its emotional state. We found that both subtypes were preferentially observed in positive contexts, but that pulsed snorts were even more associated with highly appreciated situations (in pasture and with ears forward). This study is a step further in the identification of indicators of positive emotions in horses and more generally in the understanding of the acoustic emotions' coding.

An unexpected acoustic indicator of positive emotions in horses.

Indicators of positive emotions are still scarce and many proposed behavioural markers have proven ambiguous. Studies established a link between acoustic signals and emitter's internal state, but few related to positive emotions and still fewer considered non-vocal sounds. One of them, the snort, is shared by several perrisodactyls and has been associated to positive contexts in these species. We hypothesized that this could be also the case in horses. In this species, there is a clear need for a thorough description of non-vocal acoustic signals (snorts, snores or blows are often used interchangeably) but overall this sound produced by nostrils during expiration has up to now been mostly considered as having a hygienic function. However, observations revealed that snorts were produced more in some individuals than in others, without relationship with air conditions. We observed 48 horses living in two "extreme" conditions: restricted conditions (single stall, low roughage diet) and naturalistic conditions (stable groups in pasture). The immediate place (e.g. stall/pasture) and the behavioural/postural (behaviour performed/ears positions) contexts of snort production were observed. We additionally performed an evaluation of the welfare state, using validated behavioural (e.g. stereotypies) and postural (e.g. overall ears positions) welfare indicators. The results show that 1) snort production was significantly associated with situations known to be positive for horses (e.g. feeding in pasture) and with a positive internal state (ears in forward or sidewards positions), 2) the riding school horses produced twice as many snorts when in pasture than in stall, 3) the naturalistic population emitted significantly more snorts than riding school ones in comparable contexts, 4) the frequency of snorts was negatively correlated with the composite total chronic stress score (TCSS, reflecting compromised welfare based on the horse's rank on the different indicators): the lower the TCSS, the higher the snort rate. Snorts therefore appear as reliable indicators of positive emotions.

The human newborn's umwelt: Unexplored pathways and perspectives.

Historically, newborns, and especially premature newborns, were thought to "feel nothing." However, over the past decades, a growing body of evidence has shown that newborns are aware of their environment, but the extent and the onset of some sensory capacities remain largely unknown. The goal of this review is to update our current knowledge concerning newborns' perceptual world and how ready they are to cope with an entirely different sensory environment following birth. We aim to establish not only how and when each sensory ability arises during the pre-/postbirth period but also discuss how senses are studied. We conclude that although many studies converge to show that newborns are clearly sentient beings, much is still unknown. Further, we identify a series of internal and external factors that could explain discrepancies between studies, and we propose perspectives for future studies. Finally, through examples from animal studies, we illustrate the importance of this detailed knowledge to pursue the enhancement of newborns' daily living conditions. Indeed, this is a prerequisite for assessing the effects of the physical environment and routine procedures on newborns' welfare.

Low plasma cortisol and fecal cortisol metabolite measures as indicators of compromised welfare in domestic horses (Equus caballus).

The hypothalamic-pituitary-adrenal (HPA) axis response to chronic stress is far from straight forward, particularly with regards to animal welfare. There are reports of no effect as well as both decreases and increases in cortisol after chronic stressors. Therefore, the first aim of the present study was to determine how measures of compromised welfare, such as chronic pain and haematological anomalies, related to cortisol levels in domestic horses (Equus caballus). Domestic horses are an informative model to investigate the impact of chronic stress (due to environment, pain, work, housing conditions…) on the HPA axis. The second aim was to determine whether levels of fecal cortisol metabolites (FCM) may be used as an indicator of welfare measures. The present study used fifty-nine horses (44 geldings and 15 mares), from three riding centres in Brittany, France. The primary findings show that horses whose welfare was clearly compromised (as indicated by an unusual ears backward position, presence of vertebral problems or haematological anomalies, e.g. anaemia) also had lower levels of both FCM and plasma cortisol. This work extends our previous findings showing that withdrawn postures, indicators of depressive-like behavior in horses, are associated with lower plasma cortisol levels. We also found that evening plasma cortisol levels positively correlated with FCM levels in horses. Future research aims to determine the extent to which factors of influence on welfare, such as living conditions (e.g. single stalls versus group housing in pasture or paddocks), early life factors, and human interaction, act as mediators of cortisol levels in horses.

An Ambulatory Electroencephalography System for Freely Moving Horses: An Innovating Approach.

Electroencephalography (EEG) that has been extensively studied in humans presents also a large interest for studies on animal brain processes. However, since the quality of the recordings is altered by muscular activity, most EEG recordings on animals are obtained using invasive methods with deeply implanted electrodes. This requires anesthesia and can thus only be used in laboratory or clinical settings. As EEG is a very useful tool both for detecting brain alterations due to diseases or accidents and to evaluate the arousal and attentional state of the animal, it seemed crucial to develop a tool that would make such recordings possible in the horse's home environment, with a freely moving horse. Such a tool should neither be invasive nor cause discomforts to the horse as the usual other practice which consists, after shaving the zone, in gluing the electrodes to the skin. To fulfill these requirements, we developed a novel EEG headset adapted to the horse's head that allows an easy and fast positioning of the electrodes and that can be used in the home environment on a freely moving horse. In this study, we show that this EEG headset allows to obtain reliable recordings, and we propose an original evaluation of an animal's "EEG profile" that allows comparisons between individuals and situations. This EEG headset opens new possibilities of investigation on horse cognition, and it can also become a useful tool for veterinarians to evaluate cerebral disorders or check the anesthesia level during a surgery.

Informed horses are influential in group movements, but they may avoid leading.

In species that move in cohesive groups, animals generally reach decisions through socially distributed processes, and individual knowledge is expected to influence collective decision making. Pooling of information should not be considered a general rule, however, since conflicts of interest may occur between group members. When resources are limited or highly attractive, higher-ranking individuals can prevent others from accessing food, and subordinates may have an interest in withholding information about its location. We investigated the role individual knowledge may play in recruitment processes in four groups of horses (Equus caballus). Animals were repeatedly released in a food search situation, in which one individual had been informed about the location of a preferred food, while another was a naïve control subject. Horses that were informed about the location of the feeding site were seen to approach the food source more steadily and were followed by a higher number of group members than their uninformed counterparts. Recruitment processes appeared mostly passive. Among the informed subjects, lower-ranking individuals were overall less followed than the higher-ranking ones. Most lower-ranking horses arrived alone at the feeding site. Non-followed informed subjects spent less time in active walk and used direct paths less frequently than followed animals, and they were joined by fewer partners at the attractive food source and spent more time feeding alone. This indicates that the influence of informed individuals on the behavior of other group members was a mixed process. Some horses brought nutritional benefits to their conspecifics by leading them to food supplies, whereas the behavior of others might be functionally deceptive.

Unexpected behavioural consequences of preterm newborns' clothing.

Restrictions of preterm newborns' movements could have consequences ranging from stress enhancement to impairment of their motor development. Therefore, ability to freely express motor activities appears crucial for their behavioural and physiological development. Our aim was to evaluate behavioural issues of two types of clothing used in NICU. We observed 18 healthy 34-37 post-conception week-old preterm newborns, during resting periods, when they were undisturbed by any interventions. Newborns wore either light clothing (bodysuit and a light wrapping) or heavy clothing (pyjamas, cardigan and sleep-sack). The percentages of time each subject spent in different postures were compared between clothing situations. Arm and hand postures differed in relation to clothing: babies bent their arms more and held their hands nearer their heads when in bodysuits than when in sleepwear. Consequently, babies in bodysuits spent more time touching their body or their environment whereas the others generally were touching nothing. Self-touch is an important way to comfort one's self. Heavy clothing may impair self-soothing behaviours of preterm newborn babies that already lack other forms of contact. Results suggest that more attention should be paid to apparently routine and marginal decisions such as choice of clothes.

Programming of neuroendocrine self in the thymus and its defect in the development of neuroendocrine autoimmunity.

For centuries after its first description by Galen, the thymus was considered as only a vestigial endocrine organ until the discovery in 1961 by Jacques FAP Miller of its essential role in the development of T (thymo-dependent) lymphocytes. A unique thymus first appeared in cartilaginous fishes some 500 million years ago, at the same time or shortly after the emergence of the adaptive (acquired) immune system. The thymus may be compared to a small brain or a computer highly specialized in the orchestration of central immunological self-tolerance. This was a necessity for the survival of species, given the potent evolutionary pressure imposed by the high risk of autotoxicity inherent in the stochastic generation of the diversity of immune cell receptors that characterize the adaptive immune response. A new paradigm of "neuroendocrine self-peptides" has been proposed, together with the definition of "neuroendocrine self." Neuroendocrine self-peptides are secreted by thymic epithelial cells (TECs) not according to the classic model of neuroendocrine signaling, but are processed for presentation by, or in association with, the thymic major histocompatibility complex (MHC) proteins. The autoimmune regulator (AIRE) gene/protein controls the transcription of neuroendocrine genes in TECs. The presentation of self-peptides in the thymus is responsible for the clonal deletion of self-reactive T cells, which emerge during the random recombination of gene segments that encode variable parts of the T cell receptor for the antigen (TCR). At the same time, self-antigen presentation in the thymus generates regulatory T (Treg) cells that can inhibit, in the periphery, those self-reactive T cells that escaped negative selection in the thymus. Several arguments indicate that the origin of autoimmunity directed against neuroendocrine glands results primarily from a defect in the intrathymic programming of self-tolerance to neuroendocrine functions. This defect may be genetic or acquired, for example during an enteroviral infection. This novel knowledge of normal and pathologic functions of the thymus constitutes a solid basis for the development of a novel type of tolerogenic/negative self-vaccination against type 1 diabetes (T1D).

Plasma cortisol and faecal cortisol metabolites concentrations in stereotypic and non-stereotypic horses: do stereotypic horses cope better with poor environmental conditions?

BACKGROUND: Stereotypic behaviours, i.e. repetitive behaviours induced by frustration, repeated attempts to cope and/or brain dysfunction, are intriguing as they occur in a variety of domestic and captive species without any clear adaptive function. Among the different hypotheses, the coping hypothesis predicts that stereotypic behaviours provide a way for animals in unfavourable environmental conditions to adjust. As such, they are expected to have a lower physiological stress level (glucocorticoids) than non-stereotypic animals. Attempts to link stereotypic behaviours with glucocorticoids however have yielded contradictory results. Here we investigated correlates of oral and motor stereotypic behaviours and glucocorticoid levels in two large samples of domestic horses (NStudy1 = 55, NStudy2 = 58), kept in sub-optimal conditions (e.g. confinement, social isolation), and already known to experience poor welfare states. Each horse was observed in its box using focal sampling (study 1) and instantaneous scan sampling (study 2). Plasma samples (collected in study 1) but also non-invasive faecal samples (collected in both studies) were retrieved in order to assess cortisol levels. RESULTS: Results showed that 1) plasma cortisol and faecal cortisol metabolites concentrations did not differ between horses displaying stereotypic behaviours and non-stereotypic horses and 2) both oral and motor stereotypic behaviour levels did not predict plasma cortisol or faecal cortisol metabolites concentrations. CONCLUSIONS: Cortisol measures, collected in two large samples of horses using both plasma sampling as well as faecal sampling (the latter method minimizing bias due to a non-invasive sampling procedure), therefore do not indicate that stereotypic horses cope better, at least in terms of adrenocortical activity.