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Author |
Palme, R. |
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Monitoring stress hormone metabolites as a useful, non-invasive tool for welfare assessment in farm animals |
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Journal Article |
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2012 |
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Animal Welfare |
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21 |
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3 |
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331-337 |
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animal welfare, corticosterone, cortisol, faeces, farm animals, stress |
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A multitude of endocrine mechanisms are involved in coping with challenges. Glucocorticoids, secreted by the adrenal glands, are in the front line of the battle to overcome stressful situations. They are usually measured in plasma samples as parameters of adrenal activity and thus of disturbance. Unfortunately, collecting blood samples itself can disturb an animal. Thus, non-invasive methods for the determination of glucocorticoids or their metabolites have become increasingly popular. The pros and cons of various non-invasive sample materials (saliva, excreta, milk, hair/feathers and eggs) for glucocorticoid determination are given. Above all, faecal samples offer the
advantage that they can be collected easily. In faecal samples, circulating hormone levels are integrated over a certain period of time and represent the cumulative secretion of hormones. Thus, the levels are less affected by short fluctuations or the pulse-like nature of hormone secretion. However, using this technique to assess an animal’s adrenocortical activity is not especially simple. Whether frequent sampling is necessary or single samples will suffice depends upon the study’s aim (whether one is examining the impact of acute or chronic stressors). Background knowledge of the metabolism and excretion of cortisol/corticosterone metabolites is required and a careful validation for each species and sex investigated is obligatory. The present review also addresses analytical issues regarding sample storage, extraction procedures and immunoassays and includes a comprehensive list of published studies (up to 2011) describing the use of such methods in farmed animals. Applied properly, non-invasive techniques to monitor glucocorticoid metabolites in faecal samples of various species are a useful tool for welfare assessment, especially as they are easily applied at farm or group level. |
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0962-7286 |
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Equine Behaviour @ team @ |
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5793 |
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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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Dalmau, A.; Ferret, A.; Chacon, G.; Manteca, X. |
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Title |
Seasonal Changes in Fecal Cortisol Metabolites in Pyrenean Chamois |
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Journal Article |
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2007 |
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Journal of Wildlife Management |
Abbreviated Journal |
J Wildl Manag |
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71 |
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1 |
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190-194 |
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Cadí-Moixeró, Nature Reserve, chamois, cortisol metabolites, feces, hunting reserve, Pyrenees, Rupicapra pyrenaica pyrenaica, seasonal rhythm, stress |
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We studied seasonal changes in fecal cortisol metabolites (FCM), which have been widely used as indicators of stress, in a population of Pyrenean chamois (Rupicapra pyrenaica pyrenaica) in the Cadí Range of northeastern Spain. We collected fecal samples from 2001 to 2003 in 3 particular locations with different altitudes and male or female presence, and we analyzed them for FCM and fecal nitrogen as an indicator of diet quality. We observed a clear seasonal pattern, with the highest FCM in winter, and we obtained correlations between FCM and monthly mean minimum temperatures and fecal nitrogen. We observed no effects of tourism presence, trophy hunting, or rut season on FCM. Analysis of cortisol metabolites in feces can be a good measure of winter stress in Pyrenean chamois. |
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Equine Behaviour @ team @ |
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4254 |
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Merkies, K.; McKechnie, M.J.; Zakrajsek, E. |
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Behavioural and physiological responses of therapy horses to mentally traumatized humans |
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Journal Article |
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2018 |
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Applied Animal Behaviour Science |
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Equine-assisted therapy; Ptsd; Horse; Behaviour; Cortisol; Heart rate |
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The benefits to humans of equine-assisted therapy (EAT) have been well-researched, however few studies have analyzed the effects on the horse. Understanding how differing mental states of humans affect the behaviour and response of the horse can assist in providing optimal outcomes for both horse and human. Four humans clinically diagnosed and under care of a psychotherapist for Post-Traumatic Stress Disorder (PTSD) were matched physically to four neurotypical control humans and individually subjected to each of 17 therapy horses loose in a round pen. A professional acting coach instructed the control humans in replicating the physical movements of their paired PTSD individual. Both horses and humans were equipped with a heart rate (HR) monitor recording HR every 5secs. Saliva samples were collected from each horse 30 min before and 30 min after each trial to analyze cortisol concentrations. Each trial consisted of 5 min of baseline observation of the horse alone in the round pen after which the human entered the round pen for 2 min, followed by an additional 5 min of the horse alone. Behavioural observations indicative of stress in the horse (gait, head height, ear orientation, body orientation, distance from the human, latency of approach to the human, vocalizations, and chewing) were retrospectively collected from video recordings of each trial and analyzed using a repeated measures GLIMMIX with Tukey's multiple comparisons for differences between treatments and time periods. Horses moved slower (p < 0.0001), carried their head lower (p < 0.0001), vocalized less (p < 0.0001), and chewed less (p < 0.0001) when any human was present with them in the round pen. Horse HR increased in the presence of the PTSD humans, even after the PTSD human left the pen (p < 0.0001). Since two of the PTSD/control human pairs were experienced with horses and two were not, a post-hoc analysis showed that horses approached quicker (p < 0.016) and stood closer (p < 0.0082) to humans who were experienced with horses. Horse HR was lower when with inexperienced humans (p < 0.0001) whereas inexperienced human HR was higher (p < 0.0001). Horse salivary cortisol did not differ between exposure to PTSD and control humans (p > 0.32). Overall, behavioural and physiological responses of horses to humans are more pronounced based on human experience with horses than whether the human is diagnosed with a mental disorder. This may be a reflection of a directness of movement associated with humans who are experienced with horses that makes the horse more attentive. It appears that horses respond more to physical cues from the human rather than emotional cues. This knowledge is important in tailoring therapy programs and justifying horse responses when interacting with a patient in a therapy setting. |
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0168-1591 |
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Equine Behaviour @ team @ |
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6385 |
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Author |
Simpson, B.S. |
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Title |
Neonatal foal handling |
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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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303-317 |
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Neonatal foal handling; Imprint training; Learning in foals; Heart rate; Serum cortisol concentrations |
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Recent interest has focused on the advantage of intensively handling young foals as a means of producing more tractable horses, accustomed to humans and receptive to training. To date, the effect of this intensive handling, dubbed “imprint training” in the popular literature, has not been tested. The present study compares seven foals handled intensively as neonates with eight untreated controls. The handling protocol started from 2-8 h after birth and continued daily for a total of 5 days. The protocol consisted of a series of stimuli and experiences that were each repeated until the foal no longer resisted or reacted negatively. Subsequently, foals were tested before weaning, at 4 months of age. Results indicated that handled foals (HF) ranked higher than control foals (CF) in subjective ratings of calmness (*P<0.0117) and friendliness (*P<0.0001) and in several specific handling tasks (venipuncture *P<0.0220; placing in stock *P<0.0128). Although, in approach tests all foals but one allowed approach of a person to 4 m, significantly more HF approached the person than CF (P<0.0080). In stimulus tests, foals were presented specific stimuli to which they had been tested as neonates. Two of eight CF were too unruly and dangerous to test. Of foals that could be tested, CF required significantly more time to hook-up a heart rate monitor (**P<0.0055). Split-plot analysis indicated that HF had lower heart rates to initial left-sided stimuli, presented first, than CF (*P<0.0421). In response to right-sided stimuli, heart rate scores of CF were not significantly different from HF (P<0.2259), suggesting reduced reactivity over time due to a learning effect. Behavioral responses to specific stimuli did not differ between CF and HF, suggesting that neonatal handling has a general rather than specific effect on subsequent behavior. Cortisol concentrations were measured before and after testing and the difference calculated. All foals had higher post-testing levels than pre-testing levels. There was a significant difference between HF and CF, indicating greater reactivity among the CF (*P<0.050). In general, the results indicated that foals handled as neonates were more tractable and less reactive. Specific neonatal handling tasks, such as sticking a finger up the foal's nose or patting the bottom of the foot, seemed to have no beneficial effect on related tasks such as passing a nasogastric tube or tapping with a farrier's hammer at 4 months of age. Mechanisms for the observed effect of neonatal handling require further investigation. |
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Equine Behaviour @ team @ |
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3623 |
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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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2012 |
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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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Erber, R. |
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Xenophon Publishing |
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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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Hoffmann, G.; Bockisch, F.-J.; Kreimeier, P. |
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Einfluss des Haltungssystems auf die Bewegungsaktivität und Stressbelastung bei Pferden in Auslaufhaltungssystemen |
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Journal Article |
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2009 |
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Landbauforschung – vTI Agriculture and Forestry Research |
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2 |
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59 |
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105-112 |
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Bewegung, Cortisol, Herzfrequenzvariabilität (HFV), Pferd, Verhalten [movement, cortisol, heart rate variability (HRV), horse, behaviour] |
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Frühere Untersuchungen haben gezeigt, dass die tägliche Bewegung für die Gesunderhaltung der Pferde notwendig ist. Inwieweit sich jedoch unterschiedliche Bewegungsangebote auf das Stress-und Bewegungsverhalten von Pferden in einer Gruppen-Auslaufhaltung auswirken und ob der Bewegungsbedarf der Pferde durch eine Auslaufhaltung ohne zusätzliche Bewegung gedeckt werden kann, ist der Literatur bisher nicht zu entnehmen. Daher sollte in der nachfolgend beschriebenen Untersuchung der Frage nachgegangen werden, welche Auswirkungen verschiedene Bewegungsangebote auf die Bewegungsaktivität von Pferden in Gruppen-Auslaufhaltungen haben und ob diese das Wohlbefinden der Tiere beeinflussen. Letzteres wurde durch Messung der Herzfrequenzvariabilität und Bestimmung von Cortisolmetaboliten im Pferdekot erfasst und die Bewegungsaktivität der Pferde wurde mit ALT-Pedometern bestimmt.
Verglichen wurden eine Einzel-und Gruppenhaltung mit jeweils angrenzendem Auslauf, aber ohne eine zusätzliche Bewegung der Pferde außerhalb des Stalls. In drei weiteren Varianten der Gruppenhaltung bekamen die Pferde täglichen Auslauf auf einer unbegrünten Koppel, auf einer Weide oder durch gezielte Bewegung in einer Führanlage. Die Bewegungsaktivität konnte durch die zusätzliche Bewegung in Form von Weide oder Führanlage signifikant gesteigert werden.
Ein zusätzliches Bewegungsangebot führte bei den Pferden zu einer Abnahme der Stressbelastung und sollte auch den Pferden ermöglicht werden, die in einer Gruppenhaltung gehalten werden, um ihre physische und psychische Gesundheit zu erhalten.
[Former studies confirm the necessity of daily movement for the health of a horse. But so far no description could be found in the literature how different movement offerings impact the stress and movement behaviour of horses in group husbandries with close-by discharge. The same holds true for the question whether a discharge husbandry system can meet the movement requirements of horses if there isn�t any additional movement possibility. The aim of the present study was to examine different movement offerings, their effects on the movement activities of horses in a group horse husbandry with close-by discharge and the impact of the movement on the wellbeing of the animals.
The heart rate variability and the concentration of the cortisol metabolites in the horse excrement were analyzed for detecting the wellbeing of the horses. Additionally ALT-Pedometers were used for determining the movement activity.
A single and a group husbandry system, each with closeby discharge, were compared when horses had no additional movement outside the stable. In three further variants the group husbandry was supplemented with daily time on a non-grassy pasture land, a pasture or in a horse walker. Pasture or horse walker increased movement activity significantly. Nevertheless an additional movement offering resulted in a lower stress load of the horses and should also be allowed to horses in group husbandry systems to ensure the horse�s physical and mental health.] |
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Equine Behaviour @ team @ |
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5661 |
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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 |
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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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5869 |
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Tiefenbacher, S.; Lee, B.; Meyer, J.S.; Spealman, R.D. |
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Noninvasive technique for the repeated sampling of salivary free cortisol in awake, unrestrained squirrel monkeys |
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Journal Article |
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Year |
2003 |
Publication |
American Journal of Primatology |
Abbreviated Journal |
Am. J. Primatol. |
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Volume |
60 |
Issue |
2 |
Pages |
69-75 |
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Keywords |
saliva; cortisol; squirrel monkey; sampling technique; Hpa |
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Abstract |
The use of noninvasive measures of hypothalamic-pituitary-adrenal (HPA) axis function is of growing interest among preclinical and clinical investigators. This report describes a method for the repeated assessment of salivary free cortisol in awake, unrestrained squirrel monkeys (Saimiri sciureus) based on a saliva sampling technique previously developed for rhesus monkeys. Individually housed adult male squirrel monkeys were trained to chew on dental rope attached to a pole, from which saliva was extracted by centrifugation and analyzed for cortisol by radioimmunoassay (RIA). Eight of nine monkeys readily acquired the task, reliably providing adequate saliva samples for the assay. Salivary free cortisol levels were examined in these subjects under basal conditions and in response to two types of neuroendocrine challenge. Levels of salivary free cortisol showed relatively low intra- and interindividual variability, with mean individual morning levels ranging between 17.1 and 37.9 µg/dl. Squirrel monkeys demonstrated a consistent daily rhythm in salivary free cortisol ranging from a high of 27.4 ± 5.2 µg/dl (mean ± SEM) at 12 P.M. to a low of 7.5 ± 1.6 µg/dl at 6 P.M.. Intravenous (IV) challenges with 1 µg/kg ACTH, or 10 and 50 µg/kg CRF resulted in significant increases in salivary free cortisol. The described sampling technique provides a reliable and sensitive means for repeated measurement of HPA activity in unrestrained, awake squirrel monkeys. In addition, our findings illustrate several features of HPA system rhythmicity and reactivity using salivary cortisol instead of blood plasma or serum. Am. J. Primatol. 60:69–75, 2003. © 2003 Wiley-Liss, Inc. |
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Wiley Subscription Services, Inc., A Wiley Company |
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1098-2345 |
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Equine Behaviour @ team @ |
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5573 |
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Author |
Paramastri, Y.; Royo, F.; Eberova, J.; Carlsson, H.-E.; Sajuthi, D.; Fernstrom, A.-L.; Pamungkas, J.; Hau, J. |
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Title |
Urinary and fecal immunoglobulin A, cortisol and 11-17 dioxoandrostanes, and serum cortisol in metabolic cage housed female cynomolgus monkeys (Macaca fascicularis) |
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Journal Article |
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Year |
2007 |
Publication |
Journal of Medical Primatology |
Abbreviated Journal |
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Volume |
36 |
Issue |
6 |
Pages |
355-364 |
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Keywords |
cortisol; cynomolgus monkey; immunoglobulin A; long tailed macaque; Macaca fascicularis; metabolism cage |
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Abstract |
Background and methods Quantitative enzyme-immunoassays of urinary and fecal immunoglobulin A (IgA), cortisol and 11-17-dioxoandrostanes (11,17-DOA), and serum cortisol in eight metabolic-cage-housed female cynomolgus monkeys were performed. The monkeys were divided into two groups, B and NB. Group B animals were blood sampled every 6 hours, whereas Group NB animals were not handled/blood sampled. Results No differences were recorded between the amounts of feces and urine excreted by the two groups. Group B animals excreted more urinary cortisol than did Group NB animals indicating that restraint-blood sampling resulted in a stress response. Excreted amounts of IgA and 11,17-DOA (urine and feces) did not differ between the groups. Conclusions Urinary cortisol was a reliable marker of the stress associated with repeated blood sampling. Declining amounts of excreted urinary cortisol indicated that cynomolgus monkeys acclimated quickly to repeated blood sampling in metabolism cages. Within and between animal variation in amounts of feces voided demonstrated the importance of expressing fecal markers as ‘amounts excreted per time unit per kg body weight’ rather than just measuring the concentrations in fecal samples. |
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Blackwell Publishing Ltd |
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1600-0684 |
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Equine Behaviour @ team @ |
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5854 |
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