Abstract: Non-invasive methods enable stress evaluation through measuring fecal glucocorticoid metabolites (FGMs), and immunoglobulin A (IgA) in the feces avoiding stressful blood drawing or stressful restraining of animals in the field. However, FGMs and IgA are mostly analysed in freshly frozen samples, which is difficult when fresh samples cannot be frozen immediately or frozen samples cannot be stored or transported. Good results were also derived from air-dried fecal samples, which are hampered by unstable air humidity in the field. These difficulties may be overcome, when drying of samples could be induced with colorless silica gel (SiO2) granules in a secure set-up, such as an air tight tube. We determined the speed of drying 1.5 g of a fresh fecal sample from six horses on air and on silica gel. Furthermore, FGMs and IgA were analysed in differently stored subsamples from 12 horses: in frozen fecal samples, in air- or silica gel-dried samples stored for 1 day and for 7 days, and in wet fecal samples kept in a tube at room temperature for 7 days. FGM levels remained stable in feces dried on air or on silica gel for 7 days, whereas IgA quantities showed a significant loss. Under field conditions, when freezing or transporting the frozen samples is not possible and humidity hampers air drying, drying samples on silica gel in air tight tubes appears to be very helpful and reliable for analysing FGMs.

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: The traditional way to train horses is by the application of negative reinforcement (NR). In the past few years, however, the use of positive reinforcement (PR) has become more common. To evaluate the effectiveness and the possible stressor effect of the 2 training methods, 12 horses showing severe trailer-loading problems were selected and exposed to trailer-loading. They were randomly assigned to one of the 2 methods. NR consisted of various degrees of pressure (lead rope pulling, whip tapping). Pressure was removed as soon as the horse complied. PR horses were exposed to clicker training and taught to follow a target into the trailer. Heart rate (HR) was recorded every 5 seconds and behavior denoting discomfort was observed using one-zero sampling with 10 seconds sampling intervals. Training was completed when the horse could enter the trailer upon a signal, or was terminated after a maximum of 15 sessions. Of the 12 horses, 10 reached the criterion within the 15 sessions. One horse was eliminated from the study because of illness and 1 PR horse failed to enter the trailer. A Mann-Whitney U-test indicated that the horses trained with NR displayed significantly more discomfort behavior per training session than horses trained with PR (NR: 13.26 ± 3.25; PR: 3.17 ± 8.93, P < 0.0001) and that horses in the PR group spent less time (second) per session to complete the training criterion (NR: 672.9 ± 247.12; PR: 539.81 ± 166.37, P < 0.01). A Mann-Whitney U-test showed that no difference existed in mean HR (bpm) between the 2 groups (NR: 53.06 ± 11.73 bpm; PR: 55.54 ± 6.7 bpm, P > 0.05), but a Wilcoxon test showed a difference in the PR group between the baseline of HR and mean HR obtained during training sessions (baseline PR: 43 ± 8.83 bpm; PR: 55.54 ± 6.7 bpm, P < 0.05). In conclusion, the PR group provided the fastest training solution and expressed less stress response. Thus, the PR procedure could provide a preferable training solution when training horses in potentially stressing situations.

Abstract: Abstract Conservation agencies within the European Union promote the restoration of traditional land uses as a cost-effective way to preserve biodiversity outside reserves. Although the European Union pursues the integration of the environment into strategic decision-making, it also dictates sectoral policies that may damage farmland biodiversity. We illustrate this point by outlining the socioeconomic factors that allow the persistence of traditional free-ranging horse husbandry in Galicia, northwestern Spain. Free-ranging Galician mountain ponies provide ecological and socioeconomic services including the prevention of forest fires, the maintenance of heathlands and wolves, and the attenuation of wolf-human conflicts. This traditional livestock system may have persisted because it entails negligible costs for farmers. Wolf predation upon Galician mountain ponies does not threaten farmer's economies and seems to be tolerated better than attacks to more valuable stock. Recently, European Union's regulations on animal welfare, carcass management, or meat production put new economic and administrative burdens on farmers, make free-ranging horse rearing economically unsustainable, and incentivize its abandonment. The aim of the European Union to integrate environmental policies may be successful to preserve farmland biodiversity only through careful anticipation of the side effects of apparently unrelated regulations on the fragile equilibrium that sustain traditional land uses.