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Nuñez, C. M. V., Adelman, J. S., Smith, J., Gesquiere, L. R., & Rubenstein, D. I. (2014). Linking social environment and stress physiology in feral mares (Equus caballus): Group transfers elevate fecal cortisol levels. General and Comparative Endocrinology, 196, 26–33.
Abstract: Abstract Feral horses (Equus caballus) have a complex social structure, the stability of which is important to their overall health. Behavioral and demographic research has shown that decreases in group (or band) stability reduce female fitness, but the potential effects on the physiological stress response have not been demonstrated. To fully understand how band stability affects group-member fitness, we need to understand not only behavioral and demographic, but also physiological consequences of decreases to that stability. We studied group changes in feral mares (an activity that induces instability, including both male and female aggression) on Shackleford Banks, NC. We found that mares in the midst of changing groups exhibit increased fecal cortisol levels. In addition, mares making more group transfers show higher levels of cortisol two weeks post-behavior. These results offer insights into how social instability is integrated into an animal’s physiological phenotype. In addition, our results have important implications for feral horse management. On Shackleford Banks, mares contracepted with porcine zona pellucida (PZP) make approximately 10 times as many group changes as do untreated mares. Such animals may therefore be at higher risk of chronic stress. These results support the growing consensus that links between behavior and physiological stress must be taken into account when managing for healthy, functional populations.
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Kedzierski, W., Wilk, I., & Janczarek, I. (2014). Physiological response to the first saddling and first mounting of horses: comparison of two sympathetic training methods. Animal Science Papers and Reports, 32(3), 219–228.
Abstract: There is not much research done on the influence of sympathetic training on the emotional reaction
of horses. The aim of the present study was to evaluate the emotional response and the stress level
in horses to two sympathetic training methods: (1) with the use of the “round pen technique” (RP),
and (2) in which the RP was not applied (SH). Twenty two half-bred Anglo-Arab horses (2.5
years ±3 months of age) were subject to an initial training. Eleven horses were randomly included
to the RP method and the other 11 horses for the SH method. Heart rate (HR) and saliva cortisol
concentration were measured as indicators of horse emotional arousal and stress level, respectively.
The HR values were analysed: at rest, during the habituation period, just after the first saddling
and tightening of the girth, during the first time a human leaned over the horse’s back, and during
the mounting of the horse. Saliva samples were taken before and 15 min after each training session
studied. After saddling, the HR occurred significantly higher when the RP technique was used. The
significant increase in saliva cortisol concentration was observed only after the first mounting of
the horse. Generally, the use of the RP technique did not involve more important physiological
reactions in the trained horses than did the SH method.
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Christensen, J. W., Beekmans, M., van Dalum, M., & VanDierendonck, M. (2014). Effects of hyperflexion on acute stress responses in ridden dressage horses. Physiol. Behav., 128, 39–45.
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.
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h:, M., Lévy, F., Fortin, M., Leterrier, C., & LansadLansade, L. (2013). Stress and temperament affect working memory performance for disappearing food in horses, Equus caballus. Animal Behaviour, 86(6), 1233–1240.
Abstract: In the present study, we sought to determine the influence of stress and temperament on working memory for disappearing food in horses. After assessment of five dimensions of temperament, we tested working memory of horses using a delayed-response task requiring a choice between two food locations. Delays ranging from 0 to 20 s were tested. The duration of working memory for disappearing food was first characterized without stressors (N = 26). The horses were then divided into two groups and their performance was assessed under stressful (exposure to acute stressors prior to testing, N = 12) or control conditions (N = 12). Results showed that the duration of working memory for disappearing food lasted at least 20 s under nonstressful conditions, and that under stressful conditions this duration lasted less than 12 s. This stress-induced impairment confirms in a nonrodent species that working memory performance is very sensitive to exposure to stressors. In addition, working memory performance in horses is influenced by the temperamental dimension of fearfulness according to the state of stress: fearful horses showed better performance under control conditions and worse performance under stressful conditions than nonfearful horses. These findings are discussed in the context of the Yerkes–Dodson law of stress and performance.
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Becker-Birck, M., Schmidt, A., Wulf, M., Aurich, J., von der Wense, A., Möstl, E., et al. (2013). Cortisol release, heart rate and heart rate variability, and superficial body temperature, in horses lunged either with hyperflexion of the neck or with an extended head and neck position. Journal of Animal Physiology and Animal Nutrition, 97(2), 322–330.
Abstract: Bringing the head and neck of ridden horses into a position of hyperflexion is widely used in equestrian sports. In our study, the hypothesis was tested that hyperflexion is an acute stressor for horses. Salivary cortisol concentrations, heart rate, heart rate variability (HRV) and superficial body temperature were determined in horses (n = 16) lunged on two subsequent days. The head and neck of the horse was fixed with side reins in a position allowing forward extension on day A and fixed in hyperflexion on day B. The order of treatments alternated between horses. In response to lunging, cortisol concentration increased (day A from 0.73 ± 0.06 to 1.41 ± 0.13 ng/ml, p < 0.001; day B from 0.68 ± 0.07 to 1.38 ± 0.13 ng/ml, p < 0.001) but did not differ between days A and B. Beat-to-beat (RR) interval decreased in response to lunging on both days. HRV variables standard deviation of RR interval (SDRR) and RMSSD (root mean square of successive RR differences) decreased (p < 0.001) but did not differ between days. In the cranial region of the neck, the difference between maximum and minimum temperature was increased in hyperflexion (p < 0.01). In conclusion, physiological parameters do not indicate an acute stress response to hyperflexion of the head alone in horses lunged at moderate speed and not touched with the whip. However, if hyperflexion is combined with active intervention of a rider, a stressful experience for the horse cannot be excluded.
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Zebisch, A., May, A., Reese, S., & Gehlen, H. (2013). Effect of different head-neck positions on physical and psychological stress parameters in the ridden horse. J Anim Physiol Anim Nutr, 98(5), 901–907.
Abstract: Summary Different head?neck positions (HNPs) are used in equestrian sports and are regarded as desirable for training and competition by riders, judges and trainers. Even though some studies have been indicative of hyperflexion having negative effects on horses, this unnatural position is frequently used. In the present study, the influence of different HNPs on physical and psychological stress parameters in the ridden horse was investigated. Heart rate (HR), heart rate variability (HRV) and blood cortisol levels were measured in 18 horses. Low frequency (LF) and high frequency (HF) are power components in the frequency domain measurement of HRV which show the activity of the sympathetic and parasympathetic nervous system. Values were recorded at rest, while riding with a working HNP and while riding with hyperflexion of the horse's head, neck and poll. In addition, rideability and behaviour during the different investigation stages were evaluated by the rider and by an observer. Neither the HR nor the HRV showed a significant difference between working HNP (HR = 105 ± 22/min; LF/HF = 3.89 ± 5.68; LF = 37.28 ± 10.77%) and hyperflexion (HR = 110 ± 18; LF/HF = 1.94 ± 2.21; LF = 38.39 ± 13.01%). Blood cortisol levels revealed a significant increase comparing working HNP (158 ± 60 nm) and hyperflexion (176 ± 64 nm, p = 0.01). The evaluation of rider and observer resulted in clear changes of rideability and behavioural changes for the worse in all parameters collected between a working HNP and hyperflexion. In conclusion, changes of the cortisol blood level as a physical parameter led to the assumption that hyperflexion of head, neck and poll effects a stress reaction in the horse, and observation of the behaviour illustrates adverse effects on the well-being of horses during hyperflexion.
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Palme, R., Touma, C., Arias, N., Dominchin, M.N., & Lepschy, M. (2013). Steroid extraction: Get the best out of faecal samples. Wien Tierärztl Monat – Vet Med Austria, 100, 238–246.
Abstract: Faecal steroid hormone metabolites are becoming increasingly popular as parameters for reproductive functions and stress. The extraction of the steroids from the faecal matrix represents the initial step before quantification can be performed. The steroid metabolites present in the faecal matrix are of varying polarity and composition, so selection of a proper extraction procedure is essential. There have been some studies to address this complex but often neglected point. Radiolabelled steroids (e.g. cortisol or progesterone) have frequently been added to faecal samples to estimate the efficiency of the extraction procedures used. However, native, unmetabolized steroids are normally not present in the faeces and therefore the results are artifi- cial and do not accurately reflect the actual recoveries of the substances of interest. In this respect, recovery experiments based on faecal samples from radiometabolism studies are more informative. In these samples, the metabolite content accurately reflects the mixture of metabolites present in the given species. As a result, it is possible to evaluate different extraction methods for use with faecal samples. We present studies on sheep, horses, pigs, hares and dogs that utilized samples containing naturally metabolized, 14C-labelled steroids. We recommend extracting faecal steroids by simply suspending the faeces in a high percentage of a primary alcohol (for glucocorticoid metabolites 80% aqueous methanol proved best suited for virtually all mammalian species tested so far). Not only does the procedure significantly increase the total amount of recovered radioactivity, it also increases the percentage of unconjugated metabolites, which are more likely to be recognized by the antibodies used in various immunoassays. The advantages of this extraction procedure are clear: it is very easy to use (no evaporation step is needed), it yields high recoveries and variation based on the extraction procedure is reduced to a minimum.
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Christensen, J. W. (2012). Object habituation in horses: Voluntary vs. negatively reinforced approach to frightening stimuli. In K. Krueger (Ed.), Proceedings of the 2. International Equine Science Meeting (Vol. in press). Wald: Xenophon Publishing.
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.
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Flauger, B., Möstl, E., & Krueger., K. (2012). The introduction of horses into new groups: Social interactions and cortisol release. In K. Krueger (Ed.), Proceedings of the 2. International Equine Science Meeting (Vol. in press). Wald: Xenophon Publishing.
Abstract: Domestic horses are kept in so-called “fate societies” where they have to deal with frequent mixing. Several studies have evaluated and discussed the aggression level and injury risk during the introduction of horses into new groups, but nothing is known about the endocrine responses and thus if horses experience stress during introduction.
In this study we analysed the efficiency of four approved introduction techniques and evaluated the introduction of 30 horses into 11 different groups. Horses were introduced: 1) immediately, 2) after observing the new group for several days, 3) together with an “integration horse” after several days of observation, or 4) with a mixed strategy. Aggressive as well as positive social behaviour between the introduced horses and the group members were analysed the two hours following the introduction event. In addition, we focussed on the glucocorticoid production of the newcomer horses by measuring faecal cortisol metabolites (FCM) on the day of the introduction as well as the following three days.
For the four introduction techniques we found significant differences in the horses’ aggressive and submissive behaviour as well as in their total interactions. The introduction together with an integration horse led to significantly lower levels of aggression and less total interactions than the immediate introduction of single horses.
Horses which were introduced immediately or after an observation period showed significantly elevated levels of FCM on the first, second and third day after the introduction. For horses introduced together with an integration horse FCM were already significantly higher on the day of the introduction, indicating a stressful event before the introduction itself. In contrast, FCM levels were always very low when using the mixed technique.
In sum, horses have the ability to deal with conflict when they are introduced to new group members. The introduction event itself appears not to be as stressful as previously assumed. Standing together with an “integration horse” on a separate paddock and not being able to integrate immediately into a new group appears to be stressful for the newcomer. Based on the findings of our study we suggest to introduce new horses in group management together with a new group mate, a so-called “integration horse”. This would reduce the number of total social interactions as well as the aggression level. While this technique may be stressful for the newcomer, it lowers aggressive behaviour between the introduced horse and the group members and consequently reduces injury risks.
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Hausberger, M., Fureix, C., Bourjade, M., Wessel-Robert, S., & Richard-Yris, M. - A. (2012). On the significance of adult play: what does social play tell us about adult horse welfare? Naturwissenschaften, 99(4), 291–302.
Abstract: 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.
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