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Peeters, M.; Sulon, J.; Beckers, J.-F.; Ledoux, D.; Vandenheede, M. |
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Title |
Comparison between blood serum and salivary cortisol concentrations in horses using an adrenocorticotropic hormone challenge |
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Journal Article |
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Year |
2011 |
Publication |
Equine Veterinary Journal |
Abbreviated Journal |
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43 |
Issue |
4 |
Pages |
487-493 |
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Keywords |
horse; cortisol; ACTH challenge; saliva; stress |
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Abstract  |
Reasons for performing study: In horses, serum cortisol concentration is considered to provide an indirect measurement of stress. However, it includes both free and bound fractions. The sampling method is also invasive and often stressful. This is not the case for salivary cortisol, which is collected using a more welfare-friendly method and represents a part of the free cortisol fraction, which is the biologically active form. Objectives: To compare salivary and serum cortisol assays in horses, in a wide range of concentrations, using an adrenocorticotropic hormone (ACTH) stimulation test, in order to validate salivary cortisol for stress assessment in horse. Methods: In 5 horses, blood samples were drawn using an i.v. catheter. Saliva samples were taken using swabs. Cortisol was assayed by radioimmunoassay. All data were treated with a regression method, which pools and analyses data from multiple subjects for linear analysis. Results: Mean ± s.d. cortisol concentrations measured at rest were 188.81 ± 51.46 nmol/l in serum and 1.19 ± 0.54 nmol/l in saliva. They started increasing immediately after ACTH injection and peaks were reached after 96 ± 16.7 min in serum (356.98 ± 55.29 nmol/l) and after 124 ± 8.9 min in saliva (21.79 ± 7.74 nmol/l, P<0.05). Discharge percentages were also different (225% in serum and 2150% in saliva, P<0.05). Correlation between serum and salivary cortisol concentrations showed an adjusted r2= 0.80 (P<0.001). The strong link between serum and salivary cortisol concentrations was also estimated by a regression analysis. Conclusions: The reliability of both RIAs and regression found between serum and salivary cortisol concentrations permits the validation of saliva-sampling as a noninvasive technique for cortisol level assessment in horses. |
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Blackwell Publishing Ltd |
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2042-3306 |
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Equine Behaviour @ team @ |
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5428 |
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Author |
Ayala, I.; Martos, N.F.; Silvan, G.; Gutierrez-Panizo, C.; Clavel, J.G.; Illera, J.C. |
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Title |
Cortisol, adrenocorticotropic hormone, serotonin, adrenaline and noradrenaline serum concentrations in relation to disease and stress in the horse |
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Journal Article |
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Year |
2012 |
Publication |
Research in Veterinary Science |
Abbreviated Journal |
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Volume |
93 |
Issue |
1 |
Pages |
103-107 |
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Keywords |
Horse; Disease; Cortisol; Acth; Serotonin; Catecholamines; Stress |
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Abstract |
No detailed comparative data are available on the hormonal parameters of horses suffering from a number of diseases. The aim of our study was to measure concentrations of cortisol, adrenocorticotropic hormone (ACTH), serotonin, adrenaline and noradrenaline in horses with various diseases and following surgery, to assess the response of the HPA axis and adrenal medulla. Blood samples were obtained from six groups of horses comprising a total of 119 animals as follows: laminitis, acute abdominal syndrome (AAS), castration surgery, acute diseases, chronic diseases and healthy controls. Serum hormonal concentrations were determined for each group for comparison. Statistically significant differences between all groups and controls were found for cortisol, ACTH (except for castration), serotonin and adrenaline concentrations but only in horses with laminitis and AAS for noradrenaline. No statistically significant differences were found between males and females. The largest changes in the pituitary–adrenal axis activity occurred mainly in acute diseases, laminitis and in the AAS group. |
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0034-5288 |
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Equine Behaviour @ team @ |
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5935 |
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Schmidt, A.; Biau, S.; Möstl, E.; Becker-Birck, M.; Morillon, B.; Aurich, J.; Faure, J.-M.; Aurich, C. |
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Title |
Changes in cortisol release and heart rate variability in sport horses during long-distance road transport |
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Journal Article |
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Year |
2010 |
Publication |
Domestic Animal Endocrinology |
Abbreviated Journal |
Domest Anim Endocrinol |
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38 |
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3 |
Pages |
179-189 |
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Keywords |
Horse; Transport; Cortisol; Heart rate variability |
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It is widely accepted that transport is stressful for horses, but only a few studies are available involving horses that are transported regularly and are accustomed to transport. We determined salivary cortisol immunoreactivity (IR), fecal cortisol metabolites, beat-to-beat (RR) interval, and heart rate variability (HRV) in transport-experienced horses (N = 7) in response to a 2-d outbound road transport over 1370 km and 2-d return transport 8 d later. Salivary cortisol IR was low until 60 min before transport but had increased (P < 0.05) 30 min before loading. Transport caused a further marked increase (P < 0.001), but the response tended to decrease with each day of transport. Concentrations of fecal cortisol metabolites increased on the second day of both outbound and return transports and reached a maximum the following day (P < 0.001). During the first 90 min on Day 1 of outbound transport, mean RR interval decreased (P < 0.001). Standard deviations of RR interval (SDRR) decreased transiently (P < 0.01). The root mean square of successive RR differences (RMSSD) decreased at the beginning of the outbound and return transports (P < 0.01), reflecting reduced parasympathetic tone. On the first day of both outbound and return transports, a transient rise in geometric HRV variable standard deviation 2 (SD2) occurred (P < 0.01), indicating increased sympathetic activity. In conclusion, transport of experienced horses leads to increased cortisol release and changes in heart rate and HRV, which is indicative of stress. The degree of these changes tended to be most pronounced on the first day of both outbound and return transport. |
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0739-7240 |
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Equine Behaviour @ team @ |
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5386 |
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Erber, R.; Wulf, M.; Aurich, J.; Becker-Birck, M.; Rose-Meierhöfer, S.; Möstl, E.; Hoffmann, G.; Aurich, C. |
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Title |
Physiological stress parameters in sport horse mares transferred from group housing to individual stabling |
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Conference Article |
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Year |
2012 |
Publication |
Proceedings of the 2. International Equine Science Meeting |
Abbreviated Journal |
Proc. 2. Int. Equine. Sci. Mtg |
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in press |
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horse, isolation, cortisol, heart rate, locomotion |
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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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Corporate Author |
Erber, R. |
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Xenophon Publishing |
Place of Publication |
Wald |
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Krueger, K. |
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978-3-9808134-26 |
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Equine Behaviour @ team @ |
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5542 |
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Author |
Wyss, C. |
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Title |
Does housing in a „social box“ change faecal cortisol metabolites concentration in stallions? |
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Conference Article |
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2015 |
Publication |
Proceedings of the 3. International Equine Science Meeting |
Abbreviated Journal |
Proc. 3. Int. Equine. Sci. Mtg |
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housing system, stallions, social interaction, stress, faecal cortisol metabolites |
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In order to improve the housing conditions of stallions in individual boxes by offering a possibility to have more social contact, the Swiss national stud farm tested a new box system for horses, allowing increased physical contact with the neighbouring stallion. The aim of this part of the study was to investigate whether this type of housing system (named “social box”) potentially induces a change in stress reactions in stallions compared to conventional boxes. Therefore faecal cortisol metabolite (FCM) concentration was measured as a non-invasive parameter to assess endocrine responses related to this new environment.
Four groups each consisting of eight adult Freiberger breeding stallions were included in the test design. Every stallion spent three weeks in a conventional box and in a social box respectively (cross-over design). The conventional box consisted of a separation wall with a lower opaque part and an upper part with vertical barriers (5 cm between barriers), allowing visual and olfactory contact but strongly limiting tactile contact. The separating wall of the social box consisted of two lateral sections, one part being opaque to the ceiling and the second part consisting of vertical barriers (30 cm between barriers), allowing the horse to have physical contact with its neighbour or to avoid it.
In horses, FCM concentration reflects an average level of circulating cortisol over a period of approximatively 24h. Faecal samples were collected the day following integration in social / conventional boxes, reflecting the potential stress induced by increased social interactions during the integration. In order to asses potential chronical stress, faeces samples were also collected in week one, two and three after the integration into the social / conventional box (in total: 4 samples per horse and housing system). The samples were immediately stored at -20°C until they were analysed. The samples were not analysed in the laboratory until the end of the experiment, therefore the duration of conservation in the freezer varied from 40 to 429 days.
A considerable percentage of data from groups 1 and 2 was below the detection limit (<0.8 ng/g) (Tab. 1). Thus the statistical analysis was conducted with the FCM concentration from groups 3 and 4 (n horses = 16) which contained no values below the detection limit.
Tab. 1: Details about FCM values and storage time for the 4 groups of stallions
Group Storage duration [d] Proportion of data below the detection limit (<0.8 ng/g) Mean [ng/g] Median [ng/g]
Group 1 384-429 55.6 % 2.2 0
Group 2 315-360 25.5 % 5.8 6.3
Group 3 41-79 0.0 % 8.7 8.0
Group 4 40-85 0.0 % 5.8 5.4
Despite the impressive social interactions observed between the stallions directly after being introduced into the social boxes, we did not find any differences in FCM concentration between the stallions being introduced into the conventional box and the social box on the day of integration (social box: n samples = 16, mean±SD: 6.9±4.7 ng/g; conventional box: n samples = 16, mean±SD: 9.0±11.2 ng/g; Wilcoxon signed rank test V = 70, p = 0.94).
Overall the samples taken during integration and in week one, two and three did not show evidence of changes in FCM concentration in either housing system over a longer period of time (social box: n samples = 64, mean±SD: 7.9±6.2 ng/g; conventional box: n samples = 64, mean±SD: 6.6±3.4 ng/g; Linear mixed model (LMM), p = 0.56).
Our results suggest that the possibility of having physical contact with a conspecific does not induce changes in FCM concentration in breeding stallions. The considerable percentage of values below the detection limit in groups 1 and 2 seemed to correlate with the increasing duration of storage before analysis. During the IESM Network Meeting 2015, we would like to discuss possible methodological issues and the possibilities to correctly integrate these low values in the statistical analysis. |
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Corporate Author |
Wyss, C. |
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Equine Behaviour @ team @ |
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5869 |
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Author |
Rivera, E.; Benjamin, S.; Nielsen, B.; Shelle, J.; Zanella, A.J. |
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Title |
Behavioral and physiological responses of horses to initial training: the comparison between pastured versus stalled horses |
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Journal Article |
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2002 |
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Applied Animal Behaviour Science |
Abbreviated Journal |
Appl. Anim. Behav. Sci. |
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78 |
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2-4 |
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235-252 |
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Keywords |
Horses; Housing; Training; Behavior; Stress; Cortisol; Welfare |
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Horses kept in stalls are deprived of opportunities for social interactions, and the performance of natural behaviors is limited. Inadequate environmental conditions may compromise behavioral development. Initial training is a complex process and it is likely that the responses of horses may be affected by housing conditions. Sixteen 2-year-old Arabian horses were kept on pasture (P) (n=8) or in individual stalls (S) (n=8). Twelve horses (six P and six S) were subjected to a standardized training procedure, carried out by two trainers in a round pen, and 4 horses (two P and two S) were introduced to the round pen but were not trained (C; control). On sample collection day 0, 7, 21 and 28, behavior observations were carried out, blood samples were drawn and heart rates were monitored. Total training time for the stalled horses was significantly higher than total time for the pastured horses (S: 26.4+/-1.5 min; P: 19.7+/-1.1; P=0.032). The stalled group required more time to habituate to the activities occurring from the start of training to mounting (S: 11.4+/-0.96; P: 7.3+/-0.75 min; P=0.007). Frequency of unwanted behavior was higher in the stalled horses (S: 8.0+/-2.0; P: 2.2+/-1.0; P=0.020). Pastured horses tended to have higher basal heart rates on day 0 (S: 74.7+/-4.8; P: 81.8+/-5.3 bpm; P=0.0771). While the physiological data failed to identify differences between housing groups, the behavioral data suggest that pasture-kept horses adapt more easily to training than stalled horses. |
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refbase @ user @ |
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717 |
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Cerasoli, F.; Podaliri Vulpiani, M.; Saluti, G.; Conte, A.; Ricci, M.; Savini, G.; D'Alterio, N. |
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Assessment of Welfare in Groups of Horses with Different Management, Environments and Activities by Measuring Cortisol in Horsehair, Using Liquid Chromatography Coupled to Hybrid Orbitrap High-Resolution Mass Spectrometry |
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2022 |
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Animals |
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Animals |
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12 |
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14 |
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cortisol; animal welfare; horse; Lc-Hrms/Ms |
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Horses have always been animals used for companionship, work, transportation, and performance purposes over the history of humanity; there are different ways of managing horses, but studies on how horse welfare is influenced by different activities and managements are scanty. Understanding how the management, the environment, and the different uses of horses can affect the level of stress and well-being is important not only for people associated with horses. Three groups of horses with different management, environments, and activities were selected: (1) stabled horses ridden frequently, (2) horses that perform public order service under the Italian state police, and (3) free-ranging horses. Cortisol analysis was carried out on horsehair samples using liquid chromatography coupled to hybrid orbitrap high-resolution mass spectrometry (LC-HRMS/MS), a laboratory technique used for the first time to quantify horsehair cortisol. The selection of horses to be included in the three groups was carried out by including only subjects with positive welfare assessment in accordance with the horse welfare assessment protocol (AWIN). These analyses demonstrated that the cortisol levels detected in the horsehair of free-ranging animals were significantly higher compared to those detected in stabled and working horses. These results may have been a consequence of complex environmental, managerial, and behavioral factors, which should be worth further investigation |
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Animals |
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12 |
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14 |
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2076-2615 |
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Equine Behaviour @ team @ |
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6674 |
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Flauger, B. |
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The introduction of horses into new social groups with special regard to their stress level |
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2011 |
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Pferd; Equiden; Eingliederungstechnik; Integrationspferd; Stress; Cortisol; Endokrine Reaktion; Gruppenhaltung; Verletzungsgefahr; Aggression; Futterplatzwahl; Kot; Geruchssinn; Mensch-Pferd Interaktion; horse; equids; introduction technique; integration horse; stress; cortisol; endocrine response; group housing; injury risk; aggression; feeding decision; faecal sample; olfaction; human-horse interaction |
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Horses are a highly social species living in complex social systems which should require them to memorise and generalise social experiences and distinguish between familiar and unfamiliar conspecifics. In the main part of my thesis I concentrated on the specific conflict situation of a horse being introduced into a new social group, and investigated its behaviour and stress level. Horses were either introduced (1) immediately, (2) after an observation period, or (3) together with an integration horse after an observation period. Additionally, in the second part of my thesis I arranged several experiments to elaborate additional aspects which could affect the behaviour of horses during introductions. In this study I could describe a simplified method for measuring stress through the analysis of faecal GCMs in horses. An enzyme immunoassay (EIA) for 11-oxoaetiocholanolone using 11-oxoaetiocholanolone-17-CMO: BSA (3?,11-oxo-A EIA) as antigen showed high amounts of immunoreactive substances. The new assay increases the accuracy of the test and lowers the expenses per sample; also storing of samples at room temperature after collection is less critical. This is a big advantage both in the field of wildlife management of equids and in the field of equestrian sports (chapter 1). Comparing the different introduction techniques, the introduction with an integration horse led to significantly less total interactions and lower levels of aggression than the introduction of single horses, both immediately and after several days of observing the new group. Additionally, by observing the behaviour of the horses during everyday sociality I could develop a formula describing the interrelationship between expected aggression level and enclosure size per horse. The curve takes an exponential shape. Starting from a space allowance of 300 m2 and more per horse, the amount of aggressions per hour approaches zero. For the reduction of aggression levels and injury risks in socially kept horses I recommend an enclosure size of at least 300 m2 per horse (chapter 2). I further investigated the stress level of the introduced animals. Horses which were immediately introduced did not show elevated faecal GCMs. In contrast, horses which were introduced after an observation period had slightly elevated values 2 and 3 days after the introduction. For horses introduced together with an integration horse faecal GCMs were significantly above the baseline value on the day of introduction and 1 day after it. These differences between introduction techniques indicate that the introduction event itself is not as stressful as previously assumed. Rather standing together with an integration horse and not being able to integrate immediately into the complete group elicits stress in horses (chapter 3). In the commentary of chapter 4 several studies are discussed which failed to demonstrate social learning in horses. It is argued that they did not consider important aspects which could have an influence, such as the dominance status or the social background of the horses (chapter 4). In chapter 5 a social feeding situation was investigated. The social rank as well as the position of conspecifics affected the feeding strategy of horses. Domestic horses used social cognition and strategic decision making in order to decide where to feed. When possible they tended to return to the same, continuously supplied feeding site and switched to an ?avoidance tendency? in the presence of dominant horses or when another horse was already feeding there (chapter 5). One possibility to recognize group members is through olfactory recognition. In chapter 6 it is shown that horses are able to distinguish their own from their conspecifics? faeces. In addition, they paid most attention to the faeces of those group members from which they received the highest amount of aggressive behaviour (chapter 6). Horses show cognitive abilities because they are able to use humans as local enhancement cues when searching for food, independently of their body posture or gaze consistency when the persons face them. Moreover, they seem to orientate on the attention of familiar persons more than of unfamiliar persons (chapter 7). Altogether, the results of this thesis provide further support for the view that horses show good conflict resolution strategies. They are perfectly able to deal with the conflict situation of being introduced to new group members, and the introduction event itself is not as stressful as previously assumed. It is rather suggested that standing together with an integration horse and not being able to integrate immediately into the complete group elicits stress in horses. All additional experimental set-ups could demonstrate that horses are well capable of social cognition. |
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Ph.D. thesis |
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Equine Behaviour @ team @ epub18463 |
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5736 |
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Author |
Suagee-Bedore, J.K.; Linden, D.R.; Bennett-Wimbush, K. |
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Title |
Effect of Pen Size on Stress Responses of Stall-Housed Horses Receiving One Hour of Daily Turnout |
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Journal Article |
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2021 |
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Journal of Equine Veterinary Science |
Abbreviated Journal |
J. Equine Vet. Sci. |
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98 |
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103366 |
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Agonistic behaviors; Cortisol; Group turnout; Paddock sizes |
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Group turnout provides important socializing opportunities for horses, particularly those that are primarily stalled. A high percentage of equine injuries occur during group turnout, which could partly be due to the physical constraints of fencing. To investigate appropriate paddock sizes for group turnouts, horses (n = 12) from a single herd were divided into groups of 4, stalled for 24 hours, and then turned out for 1 hour into one of three differently sized pens: 342, 263, and 184 m2 per horse. Groups rotated through pens across 3 days, receiving one treatment per day. Blood was sampled for cortisol concentrations at 08:00 hours each morning, and then at 15 and 60 minutes into the turn out sessions, and 60 minutes after return to individual stalls. Groups rotated through three turnout times: 09:00, 12:00, 14:00 hours. Counts of agonistic behaviors (chasing, contact biting, and kicking) and low-level threats (pinned ears, tail swishing, bite and kick threats) were recorded. When turned out in pens that provided 342 m2 per horse, horses exhibited reduced plasma cortisol concentrations by 15 minutes after turnout and at 1 hour after return to their stalls (P < .05). Horses in pens providing 184 m2 per horse exhibited greater agonistic (P < .001) and low-level threat (P < .01) behaviors than horses in larger pens. These data provide insight into appropriate pen sizes for horses from established herds. Providing at least 342 m2 per horse may reduce the chance of injury in horses accustomed to group turnout. |
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Equine Behaviour @ team @ |
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6694 |
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Author |
Flauger, B.; Krueger, K.; Gerhards, H.; Möstl, E. |
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Simplified method to measure glucocorticoid metabolites in faeces of horses |
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2010 |
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Veterinary Research Communications |
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Vet Res Comm |
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34 |
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2 |
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185-195 |
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ACTH challenge; enzyme immunoassay; stress behaviour; cortisol |
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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. |
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