Abstract: Endurance horses, like marathon runners, undertake sub-maximal, longterm physical exercises and in both cases thermoregulation plays a critical role. A standardised sub-maximal treadmill test was used to assess fitness and training state in endurance horses. Literature reports that trained human athletes dissipate head load better than not trained ones; in this study parameters related to thermoregulation were recorded together with other haematologic and metabolic ones. Six endurance horses, divided into two groups according to their training state, performed two submaximal tests at a distance of one week. Blood samples were collected at rest, during exercise, at the end and during recovery (10, 30 and 60 minutes after work). Heart rate (HR), respiratory rate (RR), rectal (RT) and skin (ST) temperatures were monitored through the exercise (and recovery for HR). HR resulted significantly lower (P<0.01) in trained horses than in untrained ones, especially during recovery. Also rectal and skin temperatures resulted significantly lower (P<0.05) in trained horses and ST increased more gradually during exercise. The values of haematocrit and haemoglobin resulted significantly lower (P<0.05) at rest, during exercise and recovery samples. In this study, the parameters related to thermoregulation, in accordance with data from human athletes, resulted significantly different in trained and untrained horses, while other parameters didn't vary significantly between the two groups. Thermoregulatory responses to exercise-generated heat load in a standardised test, studied by simple parameters, can thus offer useful information about fitness and training state of endurance horses.

Abstract: A human-animal relationship can be developed through subsequent interactions, affected by the positive or negative emotional valence of the proceeding one. Horses implement a process of categorization to classify humans with whom they interact as positive, negative, or neutral stimuli by evaluating the kind of approach and the nature of the contact. In these terms, human-animal interactions are emotionally charged events, eliciting specific emotional states in both subjects involved. Although the human-horse relationship has been mainly investigated through behavioral analysis, physiological indicators are needed for a more objective assessment of the emotional responses. Heart rate variability (HRV) is a commonly used autonomic nervous system (ANS) correlate estimating the sympathovagal balance as a psychophysiological marker of emotion regulation in horses. We have assumed that long-term positive relationships with humans may have a positive and immediate impact on the emotional arousal of the horse, detectable, via ANS activity, during the interaction. We analyzed horses' heartbeat dynamics during their interaction with either familiar or unfamiliar handlers, applying a standardized experimental protocol consisting of three different conditions shifting from the absence of interaction to physical contact. The ANS signals were monitored through an innovative non-invasive wearable system, not interfering with the unconscious emotional response of the animal. We demonstrated that horses appeared to feel more relaxed while physically interacting (e.g., grooming on the right side) with some familiar handlers compared to the same task performed by someone unfamiliar. The shift of the sympathovagal balance toward a vagal predominance suggests that the horses experienced a decrease in stress response as a function not only of the handler's familiarity but also of the type of interaction they are experiencing. These results constitute the objective evidence of horses' capacity to individually recognize a familiar person, adding the crucial role of familiarity with the handler as a paramount component of human-animal interaction. Our rigorous methodological approach may provide a significant contribution to various fields such as animal welfare while further investigating the emotional side of the human-animal relationships.

Abstract: For ages horses and donkeys constituted the most important domesticated animals. Even though equids were often bred and kept in close proximity to humans, surprisingly little is known about their cognitive abilities. Traditionally, horses are not regarded as clever animals; common beliefs maintain that the horses“ behaviour is merely driven by conditioned-responses. Additionally, from an anthropomorphic point of view donkeys are believed to be ”stupid" animals.
Our study investigates the equids“ ability to recover a hidden object. For this purpose the animal has to create and maintain a representation of the object and its location in space, from the moment in which it disappears from direct perception, till the moment in which it reappears. The knowledge about objects being entities that continue to exist even when they are no longer available for direct perception is refered to as the well known concept of ”object permanence".
We primarly assessed the ability of Esperia's pony and donkeys to solve a Detour problem while employing an opaque “U-shaped” barrier. Each animal observed a food bucket moving and disappearing behind the barrier. Immediately after the object"s disappearance, the animal was released to search for the object. If it solved the task by detouring the barrier it was positively reinforced.
The ability to retain in memory the hidden object as well as its spatial location was subsequently tested in the presence of two, rather than one, screens (Working Memory testing phase). The food bucket was made to move and hidden behind one of the two identical screens, while the animal was watching it. Following a pre-established delayed period of 10 sec, the animal was set free to look for the food. In such a test the detour problem is combined with the classical delayed-response task, which is in use for the comparison of memory duration in different species.
In order to recover the hidden objects, animals must encode, maintain and correctly regain from their working memory the existence of the no longer visible object and its location from their working memory.
Both donkeys and ponies performed the Detour task showing to grasp the fact that an object which is no longer perceivable still continues to exist and can be regained. They also were able to correctly retrieve the goal object after a delay of 10 s in the Working Memory tasks, showing that they had encoded, maintained and correclty retrieved from their working memory the spatial location of the hidden object as well as its existence.
Nevertheless, when Standardbreds, raised in traditional stables, were tested in identical conditions to those describe for the ponies and donkeys, they could not succeed in the Detour tasks. The reason for such differences needs to be studied Even though it would be interesting to focus on handling differences, i.e., the ponies had been living in an environment rich in natural stimuli, while the Standardbreds had lived in a man-controlled environment since birth. It is also noteworthy that, like donkeys, the Esperia's pony have a reputation for being hard to handle.
Several considerations could arise from our preliminary investigations, and we will have the pleasure to leave them open for discussion.

Abstract: Mirror Self-Recognition (MSR) is an extremely rare capacity in the animal kingdom that reveals the emergence of complex cognitive capacities (de Waal 2008). So far, MSR has been reported only in humans, chimpanzees (Gallup, 1970), bottlenose dolphins (Reiss and Marino, 2001) and Asian elephants (Plotnik et al, 2006), all species characterized by a highly developed cognition. There is growing evidence that domestic horses posses high cognitive abilities, such as cross-modal individual recognition (Proops et al, 2009), triadic post-conflict reunion to maintain social homeostasis (Cozzi et al, 2010), complex communicative systems (Whatan and McComb, 2014), flexibility in problem-solving (Lovrovich et al, 2015), and long-term memory (Hanggi and Ingersoll, 2009). All these capacities make horses a good candidate to test the ability of MSR in a domestic species. Through a classical MSR experimental paradigm (de Waal 2008) we tested eight horses living in social groups under semi-natural conditions (from the Italian Horse Protection rescue centre). Animals showing MSR typically go through four stages (Plotnik et al, 2006): (i) social response, (ii) physical mirror inspection (e.g., looking behind the mirror), (iii) repetitive mirror-testing behaviour (i.e., the beginning of mirror understanding), and (iv) self-directed behaviour (i.e., recognition of the mirror image as self). The final stage, known as the “mark-test”, is verified when a subject spontaneously uses the mirror to check for a coloured artificial mark on its own body which it cannot perceive otherwise. The horses underwent a three-phase “mark-test”: 1) with sham mark (transparent ultrasound water gel) positioned on both side at jaw level, 2) mark (yellow eye shadow mixed with ultrasound water gel) positioned on left side of jaw (with sham mark on the right), 3) mark (yellow eye shadow mixed with ultrasound water gel) positioned on right side of jaw (with sham mark on the left)
The mirror was one 0.5-cm-thick piece of 140x220-cm plexiglass glue on wood. Each test lasted one hour, horses were tested once a day, in consecutive days and at the same time. Our preliminary result on 1 horse shows some changes in self-directed behaviours which can be attributed to presence of the coloured mark. Firstly, the presence of the coloured mark significantly increased the frequency of scratching on both sides of the muzzle (p < 0.0001). The most intriguing result (p < 0.0001) comes from the comparison of the scratching rates directed towards the coloured mark side (N = 41) and the sham mark side (N = 23). Under the control condition (i.e. sham mark on both sides) no statistical difference was found for the scratching rates directed to the muzzle sides (dx N = 8; sx N = 5). Although further analyses are needed to confirm these preliminary results, our finding opens new scenarios about the evolution of Mirror Self-Recognition. The capacity of horses to recognize themselves in a mirror may be the outcome of an evolutionary convergence process driven by the cognitive pressures imposed by a complex social system and maintained despite thousands years of domestication.
Keywords:
Domestic horse · Mark test · Socio-cognitive skills · Self-awareness
References
De Waal FBM (2008) The thief in the mirror. PloS Biol 6(8):e201
Gallup GG Jr (1970) Chimpanzees: Self-recognition. Science 167: 86-87.
Reiss D, Marino L (2001). Mirror self-recognition in the bottlenose dolphin: A case of cognitive convergence. Proc Natl Acad Sci U S A 98:5937-5942.
Plotnik J, de Waal FBM, Reiss D (2006) Self-recognition in an Asian elephant. Proc Natl Acad Sci U S A 103: 17053-17057.
Proops L, McComb K, Reby D. (2009) Cross-modal individual recognition in domestic horses (Equus caballus). Proc Nat Acad Sci USA;106:947-951.
Cozzi A, Sighieri C, Gazzano A, Nicol CJ, Baragli P. Post-conflict friendly reunion in a permanent group of horses (Equus caballus). Behav Process 2010;85:185-190.
Wathan J, McComb K. The eyes and ears are visual indicators of attention in domestic horses. Curr Biol 2014;24(15): R677-R679.
Lovrovich P, Sighieri C, Baragli P (2015) Following human-given cues or not? Horses (Equus caballus) get smarter and change strategy in a delayed three choice task. Appl Anim Behav Sci, in press.
Hanggi EB, Ingersoll JF. (2009) Long-term memory for categories and concepts in horses (Equus caballus). Anim Cogn; 12:451-462.

Abstract: Mirror self-recognition (MSR), investigated in primates and recently in non-primate species, is considered a measure of self-awareness. Nowadays, the only reliable test for investigating MSR potential skills consists in the untrained response to a visual body mark detected using a reflective surface. Here, we report the first evidence of MSR at group level in horses, by facing the weaknesses of methodology present in a previous pilot study. Fourteen horses were used in a 4-phases mirror test (covered mirror, open mirror, invisible mark, visible colored mark). After engaging in a series of contingency behaviors (looking behind the mirror, peek-a-boo, head and tongue movements), our horses used the mirror surface to guide their movements towards their colored cheeks, thus showing that they can recognize themselves in a mirror. The analysis at the group level, which 'marks' a turning point in the analytical technique of MSR exploration in non-primate species, showed that horses spent a longer time in scratching their faces when marked with the visible mark compared to the non-visible mark. This finding indicates that horses did not see the non-visible mark and that they did not touch their own face guided by the tactile sensation, suggesting the presence of MSR in horses. Although a heated debate on the binary versus gradualist model in the MSR interpretation exists, recent empirical pieces of evidence, including ours, indicate that MSR is not an all-or-nothing phenomenon that appeared once in phylogeny and that a convergent evolution mechanism can be at the basis of its presence in phylogenetically distant taxa.