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Flauger, B., Krueger, K., Gerhards, H., & Möstl, E. (2010). Simplified method to measure glucocorticoid metabolites in faeces of horses. Vet Res Comm, 34(2), 185–195.
Abstract: Glucocorticoids or their metabolites can be measured in several body fluids or excreta, including plasma, saliva, urine and faeces. In recent years the measurement of glucocorticoid metabolites (GCMs) in faeces has gained increasing attention, because of its suitability for wild populations. In horses, however, the group-specific enzyme immunoassay described so far has a limited racticability due to its complex extraction procedure. Therefore, we tested the applicability of
other enzyme immunoassays for glucocorticoid metabolites. The present study clearly proved that an enzyme immunoassay (EIA) for 11-oxoetiocholanolone using 11-oxoetiocholanolone-17-CMO: BSA (3α,11-oxo-A EIA) as antigen showed high amounts of immunoreactive substances. Therefore it was possible to use just a small amount of the supernatant of a methanolic suspension of faeces. The results correlated well with the already described method for measuring GCMs in horse faeces, i.e. analysing the samples with an EIA after a two step clean up procedure of the samples (Merl et al. 2000). In addition, the 3α,11-oxo-A EIA has the advantage of providing a bigger difference between baseline values and peak values after ACTH stimulation. The new assay increased the accuracy of the test, lowered the expenses per sample, and storing samples at room temperature after collection was less critical than with other assays investigated in our study. This is a big advantage both in the field of wildlife management of equids and in the field of equestrian sports and it shows the importance of choosing an assay which is in good accordance with the metabolites excreted in a given species. Keywords: ACTH challenge; enzyme immunoassay; stress behaviour; cortisol
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Erber, R., Wulf, M., Aurich, J., Becker-Birck, M., Rose-Meierhöfer, S., Möstl, E., et al. (2012). Physiological stress parameters in sport horse mares transferred from group housing to individual stabling. In K. Krueger (Ed.), Proceedings of the 2. International Equine Science Meeting (Vol. in press). Wald: Xenophon Publishing.
Abstract: Initial equestrian training and especially first mounting of a rider are stressful challenges for young horses (1). Most young horses are raised in groups but, in association with equestrian training, they are commonly transferred to individual stabling in loose boxes. Although, in most stables, visual contact with horses in adjacent boxes is possible, separation from the herd might be an additional stressor. We have studied physiological stress parameters, in 3-year-old sport horse mares (n=8), transferred from a group stable with access to a paddock to individual boxes without paddock. Once stabled in the individual boxes, mares underwent a standard training for young horses. Horses had been accustomed to lunging and tolerating a rider on their back several weeks before the study. Mares were studied from 5 days before to 5 days after changing the stable. Cortisol concentration in saliva, locomotion activity (ALT pedometers), heart rate (HR) and HR variability (RMSSD: root mean square of successive beat-to-beat intervals) were determined. We hypothesized that the change of the stable increases cortisol release and is associated with changes in HR and RMSSD and reduced locomotion. Before mares were moved to individual boxes, cortisol concentration showed a pronounced diurnal rhythm with values around 0.6 ng/ ml in the morning and a continuous decrease throughout the day. When the mares were moved to individual boxes, cortisol concentration increased to 1.8±0.2 ng/ml and did not return to baseline values within 6 h (p<0.05 over time). On subsequent days, a diurnal rhythm was re-established but shifted to a higher level than before. Locomotion activity determined by ALT pedometers was increased for some minutes only after mares has been placed in individual boxes but was only slightly higher than during the time mares spent with the group in a paddock. On days 2-5 in individual boxes, locomotion activity was reduced compared to the group stable. HR increased and the HRV variable RMSSD decreased when mares were separated. In conclusion, separating horses during initial training from their group is an additional stressor, although the stress is less pronounced than induced by other social challenges, e.g. weaning of foals (2). When stabled in individual boxes, mares move less than when kept as a group. Horses kept in a group thus appear to exercise themselves freely, such an effect is absent when the animals are kept individually.
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Becker-Birck, M., Biau, S., Aurich, C., Möstl, E., Ille, N., & Aurich, J. (2012). Heart rate and heart rate variability in the horse and its rider: different responses to training and a public equestrian performance. In K. Krueger (Ed.), Proceedings of the 2. International Equine Science Meeting (Vol. in press). Wald: Xenophon Publishing.
Abstract: While detailed information exists on the cardiovascular response of horses to racing or endurance, much less is known about cardiovascular function of the rider in equestrian sports. Combined analysis of the horse-rider-team has not been investigated so far. In this study, we have analyzed changes in heart rate and heart variability (HRV) variables SDRR (standard deviation of beat-to-beat interval) and RMSSD (root mean square of successive beat-to-beat intervals) both in well-trained horses (n=9) and their highly experienced riders (n=7) during the airs above the ground (sauteurs en liberté of the Cadre noir de Saumur) at a public performance and at an identical, but non-public training session, both lasting for exactly 7 min. Heart rate in the horses and riders increased during the airs above the ground, both in training and in the public performance (p<0.001 over time). In the horses, this increase did not differ between training and public performance (training: from 35±6 in the stable to 97±17 beats/min during riding, performance: from 43±13 to 103±13 beats/min, respectively). In contrast, in the riders, the increase in heart rate was significantly more pronounced (p<0.01) during the public performance (basal value 91±10, maximum 150±15 beats/min) than during training (basal value 94±10, maximum 118±12 beats/min). With regard to HRV in horses, overall SDRR did not change significantly over time and did not differ significantly between training and performance. RMSSD decreased during both training and performance to the same extent, indicating a decrease in parasympathetic (vagal) tone. In the riders, both HRV variables decreased significantly during riding (p<0.001) and for SDRR the decrease was more pronounced (p<0.05) during an equestrian performance compared to a training session. During the performance SDRR decreased from a basal value of 5.0±1.5 to a minimum of 3.2±0.6 msec while respective values for the training session were 5.3±1.1 and 2.3±1.1 msec. Both a public performance and an identical training session of the airs above the ground in the ridden horse caused an increase in heart rate and a decrease in HRV variables. While increases in heart rate are mainly caused by physical activity, decreases in HRV also indicate a stress response. The cardiovascular response in the horses did not differ between a training session and a public performance but clear differences could be demonstrated in the riders. During a public performance, the increase in heart rate and decrease in SDRR were more pronounced than during a similar training session. In conclusion, the presence of an audience thus causes more pronounced sympathoadrenal activity in experienced riders than the same equestrian tasks ridden without spectators present. In contrast, the presence of an audience was without effect on sympathoadrenal activity in experienced horses. KW -
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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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Wolter, R., Pantel, N., Stefanski, V., Möstl, E., & Krueger, K. (2014). The role of an alpha animal in changing environmental conditions. Physiol. Behav., 133, 236–243.
Abstract: Abstract The maintenance and development of conservation areas by grazing of large herbivores, such as Przewalski's horses, is common practice. Several nature conservation areas house male bachelor groups of this species. When males are needed for breeding they are removed from the groups, often without considering group compositions and individual social positions. However, alpha animals are needed for ensuring group stability and decision making in potentially dangerous situations in several species. To investigate the role of the alpha male in a bachelor group, we observed the behaviour of five Przewalski's horse males during the enlargement of their enclosure. We analyzed the group's social structure and movement orders, as well as the animals' connectedness, activity budgets, and whether they moved with preferred group members and how factors such as social rank influenced the horses' behaviour. We also investigated the excretion of glucocorticoid metabolites (GCM) via faeces of the horses while exploring a new area as a parameter of glucocorticoid production. Our results show that the alpha male is important for a bachelor group in changing environmental conditions. The alpha male had the highest level of connectedness within the group. When exploring the new environment, its position in the group changed from previously being the last to being the first. Furthermore the whole group behaviour changed when exploring the new area. The stallions showed reduced resting behavior, increased feeding and did not stay close to each other. We found that the excretion of glucocorticoid metabolites of most horses rose only marginally during the first days on the new area while only the alpha male showed a significant increased amount of glucocorticoid production during the first day of the enclosure enlargement.
Keywords: Alpha male; Horse; Equus ferus przewalskii; Bachelor group; Group structure
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Wolter, R., Pantel, N., Möstl, E., & Küger, K. (2014). Das Verhalten von Przewalski Pferden in Semi-Reservaten. In Pferdetage Baden-Württemberg 2014. Stuttgart: Matthaes Medien.
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Merl, S., Scherzer, S., Palme, R., & Möstl, E. (2000). Pain causes increased concentrations of glucocorticoid metabolites in horse feces. J Equine Vet Sci, 20(9), 586–590.
Abstract: The concentration of 11,17-dioxoandrostanes (11,17-DOA), a group of cortisol metabolites, was measured using enzyme immunoassay in fecal samples of horses experiencing painful episodes. One group of horses consisted of 10 stallions castrated (samples were collected daily for 10 days); the other group was made up of 29 horses which were brought to an animal hospital because of signs of colic (samples were collected twice daily for six days). Before castration, median concentrations of 10.5 nmol/kg feces were measured. On days 1 and 2 after castration, median 11,17-DOA values increased up to 26.2 and 50.0 nmol/kg feces, respectively, and decreased thereafter to levels lower than at the beginning of the sampling period. High variations were measured between individual cases of colic. In animals with colic, all horses excreted more than 33 nmol 11,17-DOA/kg feces for various periods. The highest concentration measured was 885 nmol/kg feces. One animal out of the 29 colic horses did not show any clinical signs of pain upon arrival in the hospital. The 11,17-DOA values were below 17 nmol/kg feces in all those samples. From this data we conclude, that the concentration of 11,17-DOA in feces is a parameter for painful situations that have occurred one or two days earlier.
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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.
Keywords: animal welfare; equitation; stress; training
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Wehnert, C., Schmidt, A., Möstl, E., Aurich, J., Müller, J., & Aurich, C. (2008). Heart rate and cortisol release in horses during road transport. In IESM 2008.
Abstract: In sport horses, equestrian competitions may lead to increased cortisol release. The competition itself but also a new environment or transport to the events can be considered as stressors. In this study, warmblood sport horses (4-16 years, 4 mares, 2 stallions, 18 geldings) were transported by road for one (t1), 3.5 (t3.5) and 8 hours (t8), (n=8 per group). Heart rate (HR) is an indicator of stress and correlates with cortisol release. Heart rate was recorded with a mobile recording system (f810i, Polar, Kempele, Finland), fixed to a girth around the thorax of the horse and was monitored the day before transport, during transport and 12 hours thereafter. In addition, cortisol metabolite concentrations in faeces were determined in order to analyse adrenocortical activation during transport. Faecal samples were collected three times per day. Cortisol metabolite concentrations in faeces are increased approximately 24 hours after an elevation in plasma cortisol concentrations. Basal HR before transport did not differ between groups (45±2 (SEM) beats/min). During the first 30 min of transport, HR increased significantly (84±6, p<0.001, Kruskal-Wallis-H-test) but did not differ between groups. During the remaining part of the transport, HR was 61±6 beats/min (p<0.001 vs. first 30 min). When HR was analysed for the last 30 min of transport, values for groups t1 and t8 were significantly (p<0.05) higher than in group t3.5. HR decreased to baseline values within less than 30 min after transport and did not differ between groups (30-60 min after transport: 44±3 beats/min). Faecal cortisol metabolite concentrations on the day after transport were elevated compared to pre-transport baseline values in all three groups of horses. Significant differences existed between groups (e.g. 18 hours after transport t1: 91±5, t3.5: 110±18, t8: 136±27 ng/g; p<0.05). Based on HR analysis, horses of group t3.5 had adapted to transport, while the prolonged transport in group t8 again was a stressor. In group t1, which was transported only for one hour, the higher heart during the last 3 min of transport was still part of the initial rise in HR. After an initial increase, heart decreases indicating adaptation of the horse to transport. Cortisol metabolite concentrations in faeces indicate that transport causes adrenocortical activation with the amount of cortisol released depending on the duration of transport. Transport over medium distances appears to no major stressor and a negative impact on the performance of the horse is unlikely. Transport over 8 hours caused the most pronounced cortisol release and HR increases again during the last hours of an 8-hour transport. Transports exceeding this time may temporarily affect the subsequent performance of the horse in equestrian sports.
Supported by Stiftung Forschung für das Pferd |
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Schmidt, A., Möstl, E., Neuhauser, S., Aurich, J., Müller, J., & Aurich, C. (2008). Changes in heart rate and cortisol release during initial training of three-year-old warmblood sport horse stallions. In IESM 2008.
Abstract: The first weeks of training in young sport horses have been suggested to represent a stressful period and training practises for horses have been questioned recently. So far, only limited data on the stress response of young horses to the initial training are available. Heart rate (HR) provides information on fitness of the horse but is also an indicator of stress or pain. Determination of cortisol metabolites in faeces is a non-invasive technique to determine a prolonged stress response. In our study, three-year-old warmblood sport horse stallions (n=8) were followed through a standardised 10-week classical training programme from lunging to first mounting of a rider and progressing to moderate work. Feed, housing and management were similar for all horses. HR was recorded with a mobile recording System (f810i, Polar, Kempele, Finland) fixed to a girth around the thorax of the horse and was monitored twice weekly from 30 min before to 30 min after training, i.e. including the training period. In addition, cortisol concentrations were determined in faecal samples collected three times daily. Overall basal HR before daily training was 39±2 (SEM) beats/min and mean values did not change significantly over the 10-week study period. Average HR during initial lunging (week 1) was 119±14 beats min and decreased to 95±5 beats/min in week 2. Due to individual variations this decrease did not reach statistical significance. Neither first mounting of a rider (89±10 beats/min) nor an increasing workload (e.g. week 8: 111±4 beats/min) were associated with prolonged increases in mean HR, but transient increases were recorded and the response to mounting of the rider differed markedly between stallions. After daily training, HR decreased rapidly but was slightly, although significantly (p<0.05, Friedman-test) higher than pre-work values (46±2 beats/min). Cortisol metabolite concentrations in faeces tended to decrease during the period of lunging, were not increased when the horses were first mounted by a rider but rose slightly with an increasing work load during the last 4 weeks of the 10-week training period. In conclusion, based on HR and faecal cortisol metabolite concentrations, the initial training of sport horse stallions in the classical German training system is not associated with major stress for the horse. The increase in HR during training is due to physical exercise itself and not associated with specific situations of the training programme.
Supported by a fellowship from Stiftung Forschung für das Pferd to AS |