Exploring the similarities between risk factors triggering depression in humans and elevated in-cage "inactive but awake" behavior in laboratory mice.

Introduction: Depression is a human mental disorder that can also be inferred in non-human animals. This study explored whether time spent inactive but awake ("IBA") in the home-cage in mice was further triggered by risk factors similar to those increasing vulnerability to depression in humans (early life stress, genetic predispositions, adulthood stress). Methods: Eighteen DBA/2 J and 18 C57BL/6 J females were tested, of which half underwent as pups a daily maternal separation on post-natal days 2-14 (early-life stress "ELS") (other half left undisturbed). To assess the effect of the procedure, the time the dams from which the 18 subjects were born spent active in the nest (proxy for maternal behavior) was recorded on post-natal days 2, 6, 10 and 14 for 1 h before separation and following reunion (matched times for controls), using live instantaneous scan sampling (total: 96 scans/dam). For each ELS condition, about half of the pups were housed post-weaning (i.e., from 27 days old on average) in either barren (triggering IBA and depression-like symptoms) or larger, highly enriched cages (n = 4-5 per group). Time mice spent IBA post-weaning was observed blind to ELS treatment using live instantaneous scan sampling in two daily 90-min blocks, two days/week, for 6 weeks (total: 192 scans/mouse). Data were analyzed in R using generalized linear mixed models. Results: The dams were significantly more active in the nest over time (p = 0.016), however with no significant difference between strains (p = 0.18), ELS conditions (p = 0.20) and before/after separation (p = 0.83). As predicted, post-weaning barren cages triggered significantly more time spent IBA in mice than enriched cages (p < 0.0001). However, neither ELS (p = 0.4) nor strain (p = 0.84) significantly influenced time mice spent IBA, with no significant interaction with environmental condition (ELS × environment: p = 0.2861; strain × environment: p = 0.5713). Discussion: Our results therefore only partly support the hypothesis that greater time spent IBA in mice is triggered by risk factors for human depression. We discuss possible explanations for this and further research directions.

Do you see what I see? Testing horses' ability to recognise real-life objects from 2D computer projections.

The use of 2-dimensional representations (e.g. photographs or digital images) of real-life physical objects has been an important tool in studies of animal cognition. Horses are reported to recognise objects and individuals (conspecifics and humans) from printed photographs, but it is unclear whether image recognition is also true for digital images, e.g. computer projections. We expected that horses trained to discriminate between two real-life objects would show the same learnt response to digital images of these objects indicating that the images were perceived as objects, or representations of such. Riding-school horses (N = 27) learnt to touch one of two objects (target object counterbalanced between horses) to instantly receive a food reward. After discrimination learning (three consecutive sessions of 8/10 correct trials), horses were immediately tested with on-screen images of the objects over 10 image trials interspersed with five real object trials. At first image presentation, all but two horses spontaneously responded to the images with the learnt behaviour by contacting one of the two images, but the number of horses touching the correct image was not different from chance (14/27 horses, p > 0.05). Only one horse touched the correct image above chance level across 10 image trials (9/10 correct responses, p = 0.021). Our findings thus question whether horses recognise real-life objects from digital images. We discuss how methodological factors and individual differences (i.e. age, welfare state) might have influenced animals' response to the images, and the importance of validating the suitability of stimuli of this kind for cognitive studies in horses.

Responses to environmental enrichment are associated with personality characteristics in chestnut-bellied seed finches (Sporophila angolensis).

Environmental enrichment (EE) is used to promote natural behaviours in captive animals and may hold promise as a form of pre-release training, a strategy for improving coping skills of translocated birds. We investigated the use of EE to enhance foraging and vigilance behaviours of captive Sporophila angolensis, which may be related to post-release survival. We also evaluated whether consistent individual behavioural differences affected birds' responses to EE. We submitted 19 captive seed-finches to three short-term challenges: tonic immobility (TI), new environment (NE) and new object (NO) tests. TI behaviour is related to fear/escape response to potential predators and novelty tests (NE and NO) assess neophobia, which are ecologically relevant personality traits influencing the shyness-boldness continuum. We noted a pronounced variability among the individuals' personality traits, both in their fear and escape-related responses in the TI test and along shy/bold z-scores in NE and NO tests. During a period of enrichment, birds spent more time foraging and less time in vigilance states compared with both control phases. Personality traits of the birds affected their responses to enrichment with bolder birds spending more time foraging. The EE-related decrease in vigilance was independent of the birds' personality traits. Our findings highlight interactions between personality and rearing environment that may impact post-release outcomes for translocated animals.

Validation of complementary non-invasive tools for stress assessment in spotted paca (Cuniculus paca).

Monitoring the concentration of glucocorticoid metabolites (GCMs) in faecal samples is a non-invasive tool for physiological stress evaluation, particularly useful when studying wild species. However, both negative and positive stimuli (distress and eustress, respectively) can lead to a rise in glucocorticoids. Thus, besides validating whether GCM concentration in faeces reflects endogenous adrenal activity, we also need to identify behavioural indicators of distress to avoid misinterpretation. Therefore, we submitted four adult male spotted pacas (Cuniculus paca) to an exogenous adrenocorticotropic hormone (ACTH) challenge-test in a Latin square design (4 × 4) to monitor changes in the GCM concentration in faeces. We also aimed to describe behaviours potentially indicative of distress. We collected excreted faeces and video-recorded the animals' behaviours for five consecutive days, one day before and four days after application of the following four treatments: 1st control (no-handling); 2nd control (intra-muscular [IM] injection of saline solution); low-dose ACTH (IM injection of 0.18 ml ACTH); and high-dose ACTH (IM injection of 0.37 ml ACTH). There was a peak in the concentration of GCM in faeces collected 24 h after the injection of the high-dose ACTH treatment. Additionally, independent of the treatments, spotted pacas spent less time on exploration and feeding states, while spending more time in the inactive but awake (IBA) state following the treatment application (challenge day). The use of GCM concentration in faecal samples together with the behavioural changes (less exploration and feeding, and more IBA) showed to be efficient as a non-invasive tool for welfare assessment of farmed spotted paca.

Do greater levels of in-cage waking inactivity in laboratory mice reflect a spontaneous depression-like symptom? A pharmacological investigation.

We previously identified in laboratory mice an inactive state [being awake with eyes open motionless within the home cage; inactive but awake, 'IBA'] sharing etiological factors and symptoms with human clinical depression. We further test the hypothesis that greater time spent displaying IBA indicates a depression-like state in mice by investigating whether the antidepressant Venlafaxine, environmental enrichment, and their combination, alleviate IBA. Seventy-two C57BL/6J and 72 DBA/2J female mice were pseudo-randomly housed post-weaning in mixed strain-pairs in non-enriched (NE; 48 pairs) or in environmentally enriched (EE; 24 pairs) cages. After 34 days, half of the mice housed in NE cages were either relocated to EE cages or left in NE cages. For each of these conditions, half of the mice drank either a placebo or the antidepressant Venlafaxine (10 mg/kg). The 48 mice housed in EE cages were all relocated to NE cages and allocated to either the placebo (n = 24) or Venlafaxine (n = 24). IBA data were collected prior to and after environmental adjustment by trained observers blind to the pharmacological and environmental adjustment treatments. Data were analyzed using GLM models. NE cages triggered more IBA than EE cages (Likelihood-Ratio-Test Chi23 = 53.501, p < 0.0001). Venlafaxine and environmental enrichment appeared equally effective at reducing IBA (LRT Chi23 = 18.262, p < 0.001), and combining these approaches did not magnify their effects. Enrichment removal triggered IBA increase (LRT Chi21 = 23.050, p < 0.001), but Venlafaxine did not overcome the increase in IBA resulting from enrichment loss (LTR Chi21 = 0.081, p = 0.775). Theoretical implications for putative depression-like states in mice, and further research directions, are discussed.

Measuring affect-related cognitive bias: Do mice in opposite affective states react differently to negative and positive stimuli?

Affect-driven cognitive biases can be used as an indicator of affective (emotional) state. Since humans in negative affective states demonstrate greater responses to negatively-valenced stimuli, we investigated putative affect-related bias in mice by monitoring their response to unexpected, task-irrelevant stimuli of different valence. Thirty-one C57BL/6J and 31 DBA/2J females were individually trained to return to their home-cage in a runway. Mice then underwent an affective manipulation acutely inducing a negative (NegAff) or a comparatively less negative (CompLessNeg) affective state before immediately being tested in the runway with either an 'attractive' (familiar food) or 'threatening' (flashing light) stimulus. Mice were subsequently trained and tested again (same affective manipulation) with the alternative stimulus. As predicted, mice were slower to approach the light and spent more time with the food. DBA/2J mice were slower than C57BL/6J overall. Contrary to predictions, NegAff mice tended to approach both stimuli more readily than CompLessNeg mice, especially the light, and even more so for DBA/2Js. Although the stimuli successfully differentiated the response of mice to unexpected, task-irrelevant stimuli, further refinement may be required to disentangle the effects of affect manipulation and arousal on the response to valenced stimuli. The results also highlight the significant importance of considering strain differences when developing cognitive tasks.

Could Greater Time Spent Displaying Waking Inactivity in the Home Environment Be a Marker for a Depression-Like State in the Domestic Dog?

Dogs exposed to aversive events can become inactive and unresponsive and are commonly referred to as being "depressed", but this association remains to be tested. We investigated whether shelter dogs spending greater time inactive "awake but motionless" (ABM) in their home-pen show anhedonia (the core reduction of pleasure reported in depression), as tested by reduced interest in, and consumption of, palatable food (KongTM test). We also explored whether dogs being qualitatively perceived by experts as disinterested in the food would spend greater time ABM (experts blind to actual inactivity levels). Following sample size estimations and qualitative behaviour analysis (n = 14 pilot dogs), forty-three dogs (6 shelters, 22F:21M) were included in the main study. Dogs relinquished by their owners spent more time ABM than strays or legal cases (F = 8.09, p = 0.032). One significant positive association was found between the KongTM measure for average length of KongTM bout and ABM, when length of stay in the shelter was accounted for as a confounder (F = 3.66, p = 0.035). Time spent ABM also correlated with scores for "depressed" and "bored" in the qualitative results, indirectly suggesting that experts associate greater waking inactivity with negative emotional states. The hypothesis that ABM reflects a depression-like syndrome is not supported; we discuss how results might tentatively support a "boredom-like" state and further research directions.

Investigating determinants of yawning in the domestic (Equus caballus) and Przewalski (Equus ferus przewalskii) horses.

Yawning is rare in herbivores which therefore may be an interesting group to disentangle the potential function(s) of yawning behaviour. Horses provide the opportunity to compare not only animals living in different conditions but also wild versus domestic species. Here, we tested three hypotheses by observing both domestic and Przewalski horses living in semi-natural conditions: (i) that domestic horses may show an elevated rate of yawning as a result of the domestication process (or as a result of life conditions), (ii) that individuals experiencing a higher level of social stress would yawn more than individuals with lower social stress and (iii) that males would yawn more often than females. The study involved 19 Przewalski horses (PHs) and 16 domestic horses (DHs) of different breeds living in large outdoor enclosures. The results showed that there was no difference between the PH and DH in yawning frequency (YF). PHs exhibited much higher levels of social interactions than DHs. There was a positive correlation between yawning frequency and aggressive behaviours in PHs, especially males, supporting the idea that yawning may be associated with more excitatory/stressful social situations. A correlation was found between yawning frequency and affiliative behaviours in DHs, which supports the potential relationship between yawning and social context. Finally, the entire males, but not castrated males, showed much higher levels of yawning than females in both species. The intensity (rather than the valence) of the interaction may be important in triggering yawning, which could therefore be a displacement activity that helps reduce tension.

Stereotypic behaviour in standard non-enriched cages is an alternative to depression-like responses in C57BL/6 mice.

Depressive-like forms of waking inactivity have been recently observed in laboratory primates and horses. We tested the hypotheses that being awake but motionless within the home-cage is a depression-like symptom in mice, and that in impoverished housing, it represents an alternative response to stereotypic behaviour. We raised C57BL/6 ('C57') and DBA/2 ('DBA') females to adulthood in non-enriched (n=62 mice) or enriched (n=60 mice) cages, observing home-cage behaviour during the active (dark) phases. We predicted that being still but awake would be reduced by environmental enrichment; more pronounced in C57s, as the strain most prone to learned helplessness; negatively related to stereotypic behaviour; and positively related to immobility in Forced Swim Tests (FST). Compared to enriched mice, non-enriched subjects did spend more time spent being inactive but awake, especially if they displayed relatively little stereotypic behaviour. C57 mice also spent more time awake but motionless than DBAs. Furthermore, even after statistically controlling for housing type and strain, this behaviour very strongly tended to predict increased immobility in the FST, while high levels of stereotypic behaviours in contrast predicted low immobility in the FST. Being awake but motionless is thus a reaction to non-enriched housing that seems to be an alternative to stereotypic behaviour, and could reflect depression-like states.

Mixed-strain housing for female C57BL/6, DBA/2, and BALB/c mice: validating a split-plot design that promotes refinement and reduction.

BACKGROUND: Inefficient experimental designs are common in animal-based biomedical research, wasting resources and potentially leading to unreplicable results. Here we illustrate the intrinsic statistical power of split-plot designs, wherein three or more sub-units (e.g. individual subjects) differing in a variable of interest (e.g. genotype) share an experimental unit (e.g. a cage or litter) to which a treatment is applied (e.g. a drug, diet, or cage manipulation). We also empirically validate one example of such a design, mixing different mouse strains -- C57BL/6, DBA/2, and BALB/c -- within cages varying in degree of enrichment. As well as boosting statistical power, no other manipulations are needed for individual identification if co-housed strains are differentially pigmented, so also sparing mice from stressful marking procedures. METHODS: The validation involved housing 240 females from weaning to 5 months of age in single- or mixed- strain trios, in cages allocated to enriched or standard treatments. Mice were screened for a range of 26 commonly-measured behavioural, physiological and haematological variables. RESULTS: Living in mixed-strain trios did not compromise mouse welfare (assessed via corticosterone metabolite output, stereotypic behaviour, signs of aggression, and other variables). It also did not alter the direction or magnitude of any strain- or enrichment-typical difference across the 26 measured variables, or increase variance in the data: indeed variance was significantly decreased by mixed- strain housing. Furthermore, using Monte Carlo simulations to quantify the statistical power benefits of this approach over a conventional design demonstrated that for our effect sizes, the split- plot design would require significantly fewer mice (under half in most cases) to achieve a power of 80%. CONCLUSIONS: Mixed-strain housing allows several strains to be tested at once, and potentially refines traditional marking practices for research mice. Furthermore, it dramatically illustrates the enhanced statistical power of split-plot designs, allowing many fewer animals to be used. More powerful designs can also increase the chances of replicable findings, and increase the ability of small-scale studies to yield significant results. Using mixed-strain housing for female C57BL/6, DBA/2 and BALB/c mice is therefore an effective, efficient way to promote both refinement and the reduction of animal-use in research.

Cage-induced stereotypic behaviour in laboratory mice covaries with nucleus accumbens FosB/ΔFosB expression.

Stereotypic behaviour (SB) occurs in certain human disorders (e.g. autism), and animals treated with stimulants or raised in impoverished conditions, including laboratory mice in standard cages. Dysfunctional cortico-basal ganglia pathways have been implicated in these examples, but for cage-induced forms of SB, the relative roles of ventral versus dorsal striatum had not been fully ascertained. Here, we used immunohistochemical staining of FosB and ΔFosB to assess long-term activation within the nucleus accumbens and caudate-putamen of C57BL/6 mice. Housed in typical laboratory cages, these mice spontaneously developed different degrees of route-tracing, bar-mouthing and other forms of SB (spending 0% to over 50% of their active time budgets in this behaviour). The most highly stereotypic mice showed the most elevated FosB/ΔFosB activity in the nucleus accumbens. No such patterns occurred in the caudate-putamen. The cage-induced SB common in standard-housed mice thus involves elevated activity within the ventral striatum, suggesting an aetiology closer to compulsive gambling, eating and drug-seeking than to classic amphetamine stereotypies and other behaviours induced by motor loop over-activation.

Co-housing rodents with different coat colours as a simple, non-invasive means of individual identification: validating mixed-strain housing for C57BL/6 and DBA/2 mice.

Standard practice typically requires the marking of laboratory mice so that they can be individually identified. However, many of the common methods compromise the welfare of the individuals being marked (as well as requiring time, effort, and/or resources on the part of researchers and technicians). Mixing strains of different colour within a cage would allow them to be readily visually identifiable, negating the need for more invasive marking techniques. Here we assess the impact that mixed strain housing has on the phenotypes of female C57BL/6 (black) and DBA/2 (brown) mice, and on the variability in the data obtained from them. Mice were housed in either mixed strain or single strain pairs for 19 weeks, and their phenotypes then assessed using 23 different behavioural, morphological, haematological and physiological measures widely used in research and/or important for assessing mouse welfare. No negative effects of mixed strain housing could be found on the phenotypes of either strain, including variables relevant to welfare. Differences and similarities between the two strains were almost all as expected from previously published studies, and none were affected by whether mice were housed in mixed- or single-strain pairs. Only one significant main effect of housing type was detected: mixed strain pairs had smaller red blood cell distribution widths, a measure suggesting better health (findings that now need replicating in case they were Type 1 errors resulting from our multiplicity of tests). Furthermore, mixed strain housing did not increase the variation in data obtained from the mice: the standard errors for all variables were essentially identical between the two housing conditions. Mixed strain housing also made animals very easy to distinguish while in the home cage. Female DBA/2 and C57BL/6 mice can thus be housed in mixed strain pairs for identification purposes, with no apparent negative effects on their welfare or the data they generate. This suggests that there is much value in exploring other combinations of strains.

Comparison of clinical examinations of back disorders and humans' evaluation of back pain in riding school horses.

BACKGROUND: Questionnaires are a common tool to assess people's opinion on a large scale or to sound them out about their subjective views. The caretakers' opinion about animals' "personality" has been used in many studies. The aim of the present study was to assess whether the owners' subjective evaluation was effective to detect back disorders. Back disorders have been shown to have a high prevalence in working horses. Caretakers from 17 riding schools (1 caretaker/school, 161 horses) were given a questionnaire about their horses' health status, including back disorders. Out of these 161 horses, 59 were subjected to manual palpation of the spine and 102 were subjected to sEMG examination all along the spine. RESULTS: The results showed that subjective caretaker-reported evaluation via questionnaire survey was not efficient to detect back disorders: only 19 horses (11.8%) were reported as suffering from back pain, whereas the experimenters' evaluation detected 80 of them (49.7%) as suffering from back disorders. While most caretakers under-evaluated back disorders, a few "over-evaluated" it (more horses reported as affected than found via clinical evaluations). Horses were less prone to present back disorders when under the care of these "over-attentive" caretakers. CONCLUSIONS: This study showed that back pain is difficult to evaluate, even for professionals, and that subjective evaluations using a questionnaire is not valid in this case. The results also highlighted the real need for observational training (behaviours, postures) outside and during riding.

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.

Towards a postural indicator of back pain in horses (Equus caballus).

Postures have long been used and proved useful to describe animals' behaviours and emotional states, but remains difficult to assess objectively in field conditions. A recent study performed on horses using geometric morphometrics revealed important postural differences between 2 horse populations differing in management conditions (leisure horses living in social groups used for occasional "relaxed" riding/riding school horses living in individual boxes used in daily riding lessons with more constraining techniques). It was suggested that these postural differences may reflect chronic effects of riding techniques on the horses' kinematics and muscular development. In the present study, we tried to evaluate the interest of postural measures to assess welfare in horses. This study was separated into 2 parts. First, 18 horses coming from these 2 types of populations (leisure/riding school horses) were submitted to 2 back evaluations by 1) manual examination (experienced practitioner) and 2) sEMG measures along the spine. We then measured neck roundness on 16 of these 18 horses. The results highlighted high correlations between manual and sEMG examinations over the spine. sEMG measures at the different locations were strongly correlated all over the spine. Moreover, neck postures and muscular activities were strongly correlated, horses with concave necks having higher sEMG measures both at precise locations (i.e. cervical sites) but also when comparing neck postures to the whole spine muscular activity highlighting the functioning of horses' back as a whole. Lastly, strong differences appeared between the populations, leisure horses being evaluated as having sounder spines, exhibiting lower sEMG measures and rounder neck than the riding school horses. sEMG measures and neck "roundness" seemed therefore to be reliable indicators of back disorders, easy to evaluate in field conditions. This highlights the accuracy of using postural elements to evaluate the animals' general state and has important implications for animals' welfare evaluations.

Towards an ethological animal model of depression? A study on horses.

BACKGROUND: Recent reviews question current animal models of depression and emphasise the need for ethological models of mood disorders based on animals living under natural conditions. Domestic horses encounter chronic stress, including potential stress at work, which can induce behavioural disorders (e.g. "apathy"). Our pioneering study evaluated the potential of domestic horses in their usual environment to become an ethological model of depression by testing this models' face validity (i.e. behavioural similarity with descriptions of human depressive states). METHODOLOGY/PRINCIPAL FINDINGS: We observed the spontaneous behaviour of 59 working horses in their home environment, focusing on immobility bouts of apparent unresponsiveness when horses displayed an atypical posture (termed withdrawn hereafter), evaluated their responsiveness to their environment and their anxiety levels, and analysed cortisol levels. Twenty-four percent of the horses presented the withdrawn posture, also characterized by gaze, head and ears fixity, a profile that suggests a spontaneous expression of "behavioural despair". When compared with control "non-withdrawn" horses from the same stable, withdrawn horses appeared more indifferent to environmental stimuli in their home environment but reacted more emotionally in more challenging situations. They exhibited lower plasma cortisol levels. Withdrawn horses all belonged to the same breed and females were over-represented. CONCLUSIONS/SIGNIFICANCE: Horse might be a useful potential candidate for an animal model of depression. Face validity of this model appeared good, and potential genetic input and high prevalence of these disorders in females add to the convergence. At a time when current animal models of depression are questioned and the need for novel models is expressed, this study suggests that novel models and biomarkers could emerge from ethological approaches in home environments.

On the significance of adult play: what does social play tell us about adult horse welfare?

Play remains a mystery and adult play even more so. More typical of young stages in healthy individuals, it occurs rarely at adult stages but then more often in captive/domestic animals, which can imply spatial, social and/or feeding deprivations or restrictions that are challenging to welfare, than in animals living in natural conditions. Here, we tested the hypothesis that adult play may reflect altered welfare states and chronic stress in horses, in which, as in several species, play rarely occurs at adult stages in natural conditions. We observed the behaviour (in particular, social play) of riding school horses during occasional outings in a paddock and measured several stress indicators when these horses were in their individual home boxes. Our results revealed that (1) the number of horses and rates of adult play appeared very high compared to field report data and (2) most stress indicators measured differed between 'players' and 'non-players', revealing that most 'playful' animals were suffering from more chronic stress than 'non-playful' horses. Frequency of play behaviour correlated with a score of chronic stress. This first discovery of a relationship between adult play and altered welfare opens new lines of research that certainly deserves comparative studies in a variety of species.

Geometric morphometrics as a tool for improving the comparative study of behavioural postures.

Describing postures has always been a central concern when studying behaviour. However, attempts to compare postures objectively at phylogenetical, populational, inter- or intra-individual levels generally either rely upon a few key elements or remain highly subjective. Here, we propose a novel approach, based on well-established geometric morphometrics, to describe and to analyse postures globally (i.e. considering the animal's body posture in its entirety rather than focusing only on a few salient elements, such as head or tail position). Geometric morphometrics is concerned with describing and comparing variation and changes in the form (size and shape) of organisms using the coordinates of a series of homologous landmarks (i.e. positioned in relation to skeletal or muscular cues that are the same for different species for every variety of form and function and that have derived from a common ancestor, i.e. they have a common evolutionary ancestry, e.g. neck, wings, flipper/hand). We applied this approach to horses, using global postures (1) to characterise behaviours that correspond to different arousal levels, (2) to test potential impact of environmental changes on postures. Our application of geometric morphometrics to horse postures showed that this method can be used to characterise behavioural categories, to evaluate the impact of environmental factors (here human actions) and to compare individuals and groups. Beyond its application to horses, this promising approach could be applied to all questions involving the analysis of postures (evolution of displays, expression of emotions, stress and welfare, behavioural repertoires…) and could lead to a whole new line of research.

Partners with bad temper: reject or cure? A study of chronic pain and aggression in horses.

BACKGROUND: Experiencing acute pain can affect the social behaviour of both humans and animals and can increase the risk that they exhibit aggressive or violent behaviour. However, studies have focused mainly on the impact of acute rather than chronic painful experiences. As recent results suggest that chronic pain or chronic discomfort could increase aggressiveness in humans and other mammals, we tested here the hypothesis that, in horses, aggression towards humans (a common source of accidents for professionals) could be linked to regularly reported vertebral problems of riding horses. METHODOLOGY/PRINCIPAL FINDINGS: Vertebral examination and standardized behavioural tests were made independently on the same horses. Here we showed that most horses severely affected by vertebral problems were prone to react aggressively towards humans (33/43 horses, chi-square test, df=1, χ(2)=12.30, p<0.001), which was not the case for unaffected or slightly affected horses (9/16 horses, chi-square test, df=1, χ(2)=0.25, p>0.05). The more affected they were, the fewer positive reactions they exhibited (rs=-0.31, p=0.02). CONCLUSIONS/SIGNIFICANCE: This is to our knowledge the first experimental evidence of such a link between chronic discomfort/potential pain (inferred from the presence of vertebral problems) and aggression, suggesting that chronic painful experiences may act in ways similar to those of acute experiences. Chronic discomfort or pain may often be overlooked when facing "bad tempered" individuals, whether humans or animals. This experimental study confirms the importance of including chronic discomfort or pain as a major factor in interpersonal relations and models of aggression.

Human direct actions may alter animal welfare, a study on horses (Equus caballus).

BACKGROUND: Back pain is the cause of bad welfare in humans and animals. Although vertebral problems are regularly reported on riding horses, these problems are not always identified nor noticed enough to prevent these horses to be used for work. METHODOLOGY/PRINCIPAL FINDINGS: Nineteen horses from two riding centres were submitted to chiropractic examinations performed by an experienced chiropractor and both horses' and riders' postures were observed during a riding lesson. The results show that 74% of horses were severely affected by vertebral problems, while only 26% were mildly or not affected. The degree of vertebral problems identified at rest was statistically correlated with horses' attitudes at work (neck height and curve), and horses' attitudes at work were clearly correlated with riders' positions. Clear differences appeared between schools concerning both riders' and horses' postures, and the analysis of the teachers' speech content and duration highlighted differences in the attention devoted to the riders' position. CONCLUSION/SIGNIFICANCE: These findings are to our knowledge the first to underline the impact of riding on horses' back problems and the importance of teaching proper balance to beginner riders in order to increase animals' welfare.