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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 |
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Volume |
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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Aurich, J.; Wulf, M.; Ille, N.; Erber, R.; von Lewinski, M.; Palme, R.; Aurich, C. |
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Title |
Effects of season, age, sex and housing on salivary cortisol concentrations in horses |
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Journal Article |
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Year |
2015 |
Publication |
Domestic Animal Endocrinology |
Abbreviated Journal |
Domest. Anim. Endocrinol. |
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horse; cortisol; diurnal rhythm; reproduction; housing |
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Abstract Analysis of salivary cortisol is increasingly used to assess stress responses in horses. Since spontaneous or experimentally induced increases in cortisol concentrations are often relatively small for stress studies proper controls are needed. This requires an understanding of factors affecting salivary cortisol over longer times. In this study, we have analysed salivary cortisol concentration over 6 mo in horses (n = 94) differing in age, sex, reproductive state and housing. Salivary cortisol followed a diurnal rhythm with highest concentrations in the morning and a decrease throughout the day (P < 0.001). This rhythm was disrupted in individual groups on individual days; however, alterations remained within the range of diurnal changes. Comparison between months showed highest cortisol concentrations in December (P < 0.001). Cortisol concentrations increased in breeding stallions during the breeding season (P < 0.001). No differences in salivary cortisol concentrations between non-pregnant mares with and without a corpus luteum existed. In stallions, mean daily salivary cortisol and plasma testosterone concentration were weakly correlated (r = 0.251, P < 0.01). No differences in salivary cortisol between female and male young horses and no consistent differences between horses of different age existed. Group housing and individual stabling did not affect salivary cortisol. In conclusion, salivary cortisol concentrations in horses follow a diurnal rhythm and are increased in active breeding sires. Time of the day and reproductive state of the horses are thus important for experiments that include analysis of cortisol in saliva. |
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0739-7240 |
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Equine Behaviour @ team @ |
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5847 |
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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 |
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93 |
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1 |
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103-107 |
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Keywords |
Horse; Disease; Cortisol; Acth; Serotonin; Catecholamines; Stress |
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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.; Aurich, J.; Möstl, E.; Müller, J.; Aurich, C. |
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Title |
Changes in cortisol release and heart rate and heart rate variability during the initial training of 3-year-old sport horses |
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Journal Article |
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Year |
2010 |
Publication |
Hormones and Behavior |
Abbreviated Journal |
Horm Behav |
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58 |
Issue |
4 |
Pages |
628-636 |
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Keywords |
Horse; Initial training; Cortisol; Heart rate variability |
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Based on cortisol release, a variety of situations to which domestic horses are exposed have been classified as stressors but studies on the stress during equestrian training are limited. In the present study, Warmblood stallions (n = 9) and mares (n = 7) were followed through a 9 respective 12-week initial training program in order to determine potentially stressful training steps. Salivary cortisol concentrations, beat-to-beat (RR) interval and heart rate variability (HRV) were determined. The HRV variables standard deviation of the RR interval (SDRR), RMSSD (root mean square of successive RR differences) and the geometric means standard deviation 1 (SD1) and 2 (SD2) were calculated. Nearly each training unit was associated with an increase in salivary cortisol concentrations (p < 0.01). Cortisol release varied between training units and occasionally was more pronounced in mares than in stallions (p < 0.05). The RR interval decreased slightly in response to lunging before mounting of the rider. A pronounced decrease occurred when the rider was mounting, but before the horse showed physical activity (p < 0.001). The HRV variables SDRR, RMSSD and SD1 decreased in response to training and lowest values were reached during mounting of a rider (p < 0.001). Thereafter RR interval and HRV variables increased again. In contrast, SD2 increased with the beginning of lunging (p < 0.05) and no changes in response to mounting were detectable. In conclusion, initial training is a stressor for horses. The most pronounced reaction occurred in response to mounting by a rider, a situation resembling a potentially lethal threat under natural conditions. |
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0018-506x |
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Equine Behaviour @ team @ |
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5223 |
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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 |
Issue |
3 |
Pages |
179-189 |
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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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Author |
Schmidt, A.; Möstl, E.; Wehnert, C.; Aurich, J.; Müller, J.; Aurich, C. |
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Title |
Cortisol release and heart rate variability in horses during road transport |
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Journal Article |
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Year |
2010 |
Publication |
Hormones and Behavior |
Abbreviated Journal |
Horm. Behav. |
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57 |
Issue |
2 |
Pages |
209-215 |
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Keywords |
Horse; Transport; Cortisol; Heart rate variability |
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Based on plasma cortisol concentrations it is widely accepted that transport is stressful to horses. So far, cortisol release during transport has not been evaluated in depth by non-invasive techniques such as analysis of salivary cortisol and faecal cortisol metabolites. Transport also causes changes in heart rate and heart rate variability (HRV). In this study, salivary cortisol, faecal cortisol metabolites, heart rate and HRV in horses transported by road for short (one and 3.5 h) and medium duration (8 h) were determined. With the onset of transport, salivary cortisol increased immediately but highest concentrations were measured towards the end of transport (4.1 ± 1.6, 4.5 ± 2.6, 6.5 ± 1.8 ng/ml in horses transported for one, 3.5 and 8 h, respectively). Faecal cortisol metabolite concentrations did not change during transport, but 1 day after transport for 3.5 and 8 h had increased significantly (p < 0.01), reflecting intestinal passage time. Compared to salivary cortisol, changes in faecal cortisol metabolites were less pronounced. Heart rate increased and beat-to-beat (RR) interval decreased (p < 0.05) with the onset of transport. Standard deviation of heart rate increased while root mean square of successive RR differences (RMSSD) decreased in horses transported for 3.5 (from 74 ± 5 to 45 ± 6 ms) and 8 h (from 89.7 ± 7 to 59 ± 7 ms), indicating a reduction in vagal tone. In conclusion, transport of horses over short and medium distances leads to increased cortisol release and changes in heart rate and HRV indicative of stress. The degree of these changes is related to the duration of transport. Salivary cortisol is a sensitive parameter to detect transient changes in cortisol release. |
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0018-506x |
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Equine Behaviour @ team @ |
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5387 |
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Stull, C.L.; Spier, S.J.; Aldridge, B.M.; Blanchard, M.; Stott, J.L. |
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Immunological response to long-term transport stress in mature horses and effects of adaptogenic dietary supplementation as an immunomodulator |
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2004 |
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Equine Veterinary Journal |
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36 |
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7 |
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583-589 |
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horse; transportation; Cd+; lymphocytes; stress; cortisol; adaptogens |
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Reasons for performing study: Little information exists on the immunological effects of transport or the use of supplements to minimise transport stress. Objectives: To establish baseline ranges and evaluate immunophenotypic and functional changes associated with transport and a nutritional ‘adaptogen’ supplement. Methods: Horses received either supplement (n = 10) or placebos (n = 9) during the 30 day study. After 28 days in stalls, 12 horses (6 supplement; 6 placebo) were transported for 24 h, then unloaded and recovered. Venous blood samples were collected on Days 1, 14 and 28 to establish baselines, and on Days 28, 29 and 30 to examine changes during transport and recovery. Results: Transport prompted elevations (P<0.05) in cortisol concentration, neutrophil count and white blood cell counts, while lymphocyte subpopulation counts (CD3+, CD4+, CD8+, CD21+) decreased (P<0.05). Normal phenotypic lymphocyte profiles returned within 24 h of recovery. Supplement effects on immunophenotype (CD21+ and CD8+) were observed in stabled horses (P<0.05), but not in transported horses. Conclusions: These results provide insights into the immunological mechanisms associated with long-term transport. Potential relevance: The existence of a small window of immunological uncertainty follows long-term transportation, enhancing the potential risk of infectious disease in susceptible individuals. |
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Blackwell Publishing Ltd |
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2042-3306 |
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Equine Behaviour @ team @ |
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5845 |
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Rivera, E.; Benjamin, S.; Nielsen, B.; Shelle, J.; Zanella, A.J. |
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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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Anderson, M.K.; Friend, T.H.; Evans, J.W.; Bushong, D.M. |
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Title |
Behavioral assessment of horses in therapeutic riding programs |
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Journal Article |
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1999 |
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Applied Animal Behaviour Science |
Abbreviated Journal |
Appl. Anim. Behav. Sci. |
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63 |
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1 |
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11-24 |
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Horses; Therapeutic riding; Temperament; Cortisol; Catecholamines |
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A behavioral assessment of horses who were being used and not used in therapeutic riding programs was conducted to help determine useful methods of selecting horses for use in therapeutic riding programs. A total of 103 horses (76 horses from five therapeutic riding centers and 27 non-therapeutic riding horses from four sites) were used. Temperament survey for each horse were completed by three riding instructors at each therapeutic riding center or by the individual most knowledgeable about the horse at the other sites. Twenty personality traits from the survey were used to quantify temperament. Concentrations of plasma cortisol, norepinephrine and epinephrine were also measured in each horse. A reactivity test was then conducted which involved introducing three novel stimuli: a walking and vocalizing toy pig placed on a cardboard surface in front of the horse for 20 s; popping a balloon near the horse's flank area; and suddenly opening an umbrella and holding it open in front of the horse for 20 s. Reactions (expressions, vocalizations and movement) to each of the stimuli were scored and used to calculate an average reactivity score for each horse. The therapeutic riding instructors did not often agree on the temperament of their center's horses. The personality trait ratings made by the therapeutic riding instructors at each center were on average significantly correlated (P<0.01, r>0.52) for only 37.8% of the horses for any two instructors and 7.8% for three instructors. No significant correlations were found between temperament, reactivity, and the hormone concentrations (r<0.19), but regression analysis indicated a possibility of predicting temperament from the reactivity score and hormone concentrations (P<0.08). There was also a tendency for relationships between extremes in temperament (desirable vs. undesirable) and the hormone concentrations (P<0.09), and between extremes in reactivity (low vs. high) and the hormone concentrations (P=0.08). The difference in ratings among riding instructors indicates a need for more collaboration between instructors when evaluating horse temperament. This study also indicates that it was very difficult to objectively determine the suitability of horses for therapeutic riding programs regarding their temperament and reactivity, probably because other traits (e.g., smoothness of gait) are also considered very important. |
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0168-1591 |
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Equine Behaviour @ team @ |
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4812 |
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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 |
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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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Wyss, C. |
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Equine Behaviour @ team @ |
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