Abstract: The ability and ease of horses to habituate to frightening stimuli greatly increases safety in the horse-human relationship. Several different techniques have been suggested for habituation training of horses and under certain conditions, preventing animals from avoidance reactions during exposure to frightening stimuli is believed to facilitate habituation. Response prevention does, however, lead to a loss of control, which is a known stress inducer in both animals and humans. This experiment investigated whether horses show increased stress responses when negatively reinforced to approach a mildly frightening stimulus, compared to horses allowed to voluntarily explore the same stimulus. We further investigated whether the prevention of avoidance responses in horses that are negatively reinforced to approach the stimulus, facilitates habituation to the stimulus. Twenty-two 2-3 years old Danish warmblood geldings were included in the study. Half of the horses (NR group) were negatively reinforced (through halter and rope pressure) by a familiar human handler to approach a collection of frightening objects (six open and colourful umbrellas) placed in a semi-circle in a familiar test arena. The other half of the horses were released in the arena and were free to avoid or explore the objects (VOL group). On the next day, all horses were exposed to the objects again without a human to investigate the rate of habituation. Behavioural and heart rate responses were recorded on both days. Data were analysed in a two way repeated measures ANOVA and post hoc analysed via the Holm-Sidak method. In the VOL group, all horses initially chose to avoid the unknown objects, whereas the handler managed to get all horses in the NR group to approach and stand next to the objects within the first 2-min session. As expected, horses in the NR group had a significantly longer duration of alertness (sec, mean ± se: NR: 23 ± 4.1 vs. VOL: 16 ± 4.7, P=0.026) and a higher max HR in the first session (bpm, mean ± se: NR: 106 ± 5.2 vs. VOL: 88 ± 4.4, P=0.004). On the next day, however, the NR horses spent significantly less time investigating the objects (sec, mean ± se: NR: 13 ± 4.1 vs. VOL: 24 ± 6.0, P=0.005) and had a shorter latency to approach a feed container, placed next to the objects (sec, mean ± se: NR: 25 ± 3.9 vs. VOL: 47 ± 16.2, P=0.031), indicating increased habituation. In conclusion, negatively reinforced approach to mildly frightening objects appears to increase stress responses during the initial exposure, but also to facilitate habituation in young horses.

Abstract: The aim of the study was to investigate whether heart rate variability (HRV) could assess alterations of the autonomic nervous system (ANS) at different levels of excitement. The behavioural and physiological responses of 20 warmblood horses to a challenging ground exercise task were studied. Prior to the experiment, the horses were evaluated at rest and during forward walking (FW). The horses were then forced to move backwards continuously during 3 min according to a standardised protocol (BW1). Subsequently, the horses were exposed to two training sessions, after which the backward walking (BW2) was re-evaluated. Heart rate (HR) and HRV-parameters such as the standard deviation of the beat-to-beat intervals (SDRR), the low (LF; sympathetic tone) and high frequency (HF) component of HRV (HF; parasympathetic tone) and their ratio (LF/HF; index representing the sympatho-vagal balance) were sampled at rest, and during FW, BW1 and BW2. Stress-related behaviour during BW1 and BW2 was determined from video recordings. The results of the different evaluations were compared to each other. Compared to rest and FW, the first backward experiment induced a significant rise in HR, LF and LF/HF and a significant decrease of HF. SDRR decreased from both FW and rest with only the latter reaching significance. In BW2 after the training sessions, HR and the parameters of the sympathetic branch of the ANS (LF, LF/HF) were decreased and the vagal tone (HF) increased compared to BW1; all changes were significant. The duration of stress indicating behavioural patterns revealed also a significant decrease of excitement after the training, when backward walking did not differ from forward walking in any parameter. Correlations between HRV-parameters and stress indicating behaviour as well as HR were found. We conclude that the HRV-parameters LF and HF are valuable measures to quantify sympatho-vagal balance, which allows a more precise assessment of the responses of HR and SDRR to mental stress during low intensity exercise.

Abstract: The continuous development of tourism and related leisure activities is exerting an increasingly intense pressure on wildlife. In this study, a novel noninvasive method for measuring stress in the black grouse, an endangered, emblematic species of European ecosystems that is currently declining in several parts of its European range, is tested and physiologically validated. A radiometabolism study and an ACTH challenge test were performed on four captive black grouse (two of each sex) in order to get basic information about the metabolism and excretion of corticosterone and to find an appropriate enzyme-immunoassay (EIA) to measure its metabolites in the feces. Peak radioactivity in the droppings was detected within 1 to 2 hours. Injected (3)H-corticosterone was excreted as polar metabolites and by itself was almost absent. A cortisone-EIA was chosen from among seven tested EIAs for different groups of glucocorticoid metabolites, because it cross-reacted with some of the formed metabolites and best reflected the increase of excreted corticosterone metabolites, after the ACTH challenge test. Concentrations of the metabolites from fecal samples collected from snow burrows of free-ranging black grouse were within the same range as in captive birds. The noninvasive method described may be appropriate for evaluating the stress faced by free-living black grouse populations in the wild, particularly in mountain ecosystems where human disturbance, especially by winter sports, is of increasing conservation concern.

Abstract: The effects of hyperflexion on the welfare of dressage horses have been debated. This study aimed to investigate acute stress responses of dressage horses ridden in three different Head-and-Neck-positions (HNPs). Fifteen dressage horses were ridden by their usual rider in a standardised 10-min dressage programme in either the competition frame (CF), hyperflexion (“Low-Deep-and-Round”; LDR) or a looser frame (LF) in a balanced order on three separate test days. Heart rate (HR), heart rate variability parameters (HRV), behaviour and rein tension were recorded during the test. Salivary cortisol concentrations were measured 60min before and 0, 5, 15 and 30min after the test. Rein tension was significantly lower in LF and did not differ between CF and LDR; however approx. 15% of recordings in CF and LDR were above the sensor detection limit of 5kg. The horses had significantly higher cortisol concentrations directly after LDR compared to LF. In addition, the horses showed more distinctive head movements, including head waving, during LDR. There were no significant treatment effects on HR and HRV. In conclusion, the results indicate that LDR may be more stressful to these horses during riding.

Abstract: The first month of training of a young horse is suspected to be stressful, but the endocrine responses to initial training are unknown. Therefore in our study a total of 40 Quarter Horses (QH), all at the age of almost 2 years, were followed during the first 30 days of their training. During this time faecal samples were collected twice daily and faecal cortisol metabolites (FCM) were measured. Baseline values of FCM ranged between 1.3 and 20.1 (median: 6.7) ng/g faeces. No differences in FCM values between days of training were found. Mares showed the highest values. Significant diurnal variations were observed in mares (p=0.035) and stallions (p=0.003), but not in geldings (p=0.282). As in this study adrenocortical activity was not increased during initial training, horses seem to cope very well with this new situation. The results of our large-scale study provide basic physiological data about initial training. This gives additional input in an emotional debate about animal welfare aspects of first time handling and training of horses.
Abbreviations: 11,17-DOA = 11,17-dioxoandrostanes; EIA = Enyzme Immunoassay; FCM = faecal cortisol metabolites; GC = glucocorticoids; HPA-axis = hypothalamic-pituitary-adrenocortical-axis; QH = Quarter Horses
[Das Einreiten eines jungen Pferdes steht unter Verdacht belastend zu sein. Bisher gibt es aber keine Veröffentlichungen über endokrine Vorgänge während dieser Phase. Mit der vorliegenden Studie wurde überprüft, ob Pferde aufgrund physischer und psychischer Belastungen während des Trainings höhere Konzentrationen an Kortisolmetaboliten im Kot (FCM) aufweisen. Es wurden dazu 40 Quarter Horses im Alter von 2 Jahren während der ersten 30 Tage der Grundausbildung des Westernreitens beobachtet und ihre FCM Werte gemessen. Während dieser Zeitspanne wurden täglich morgens und abends Kotproben der Pferde genommen. Die Basalwerte der FCM Konzentration variierten zwischen 1,3 und 20,1 (Median: 6,7) ng/g Kot, wobei Stuten die höchsten Werte hatten. Signifikante Unterschiede während der einzelnen Trainingstage konnten nicht festgestellt werden. In der Tagesrhythmik wurden signifikante Unterschiede bei Stuten (p=0,035) und bei Hengsten (p=0,003), jedoch nicht bei Wallachen (p=0,282) ermittelt. In dieser Studie konnte keine erhöhte Aktivität der Nebennierenrinde im Verlauf der Grundausbildung eines Pferdes im Westernreitstil festgestellt werden. Das legt nahe, dass Pferde mit dieser neuen, zeitlich kurz andauernden Situationen gut zurechtkommen. Unsere Studie wurde an einer großen Anzahl von Tieren unter Feldbedingungen durchgeführt. Sie bietet daher eine gute Datenbasis über Belastungen während des Einreitens. Damit liefert sie einen zusätzlichen Beitrag zu einer mitunter emotional geführten Debatte über tierschutzrelevante Aspekte bei der Grundausbildung von Pferden.]