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Cerasoli, F.; Podaliri Vulpiani, M.; Saluti, G.; Conte, A.; Ricci, M.; Savini, G.; D'Alterio, N. |
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Title |
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 |
Abbreviated Journal |
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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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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Year |
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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0737-0806 |
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Equine Behaviour @ team @ |
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6694 |
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Palme, R. |
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Title |
Non-invasive measurement of glucocorticoids: Advances and problems |
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Journal Article |
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2019 |
Publication |
Physiology & Behavior |
Abbreviated Journal |
Physiol. Behav. |
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199 |
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229-243 |
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Glucocorticoids; Non-invasive; Faecal cortisol/corticosterone metabolites; Immunoassays; Physiological/biological validation |
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Glucocorticoids (GCs; i.e. cortisol/corticosterone) are a central component of the stress response and thus their measurement is frequently used to evaluate the impact of stressful situations. Their metabolites from faeces of various animal species are more and more taken as a non-invasive aid to assess GC release and thus adrenocortical activity. The current literature review includes an extensive collection (1327 papers) and evaluation (see also Supplementary Tables) of the literature on faecal cortisol/corticosterone metabolite (FCM) analysis published to date. It aims at giving reference for researchers interested in implementing FCM analysis into their study or seeking to improve such methods by providing background knowledge on GC metabolism and excretion, conveying insights into methodological issues and stating caveats of FCM analysis and by highlighting prerequisites for and some examples of a successful application of such methods. Collecting faecal samples and analysing FCMs may appear simple and straightforward, but researchers have to select and apply methods correctly. They also need to be aware of the many pitfalls and potentially confounding factors and, last but not least, have to carefully interpret results. Applied properly, measurement of FCMs is a powerful non-invasive tool in a variety of research areas, such as (stress) biology, ethology, ecology, animal conservation and welfare, but also biomedicine. |
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0031-9384 |
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Palme2019_attachment.pdf |
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Equine Behaviour @ team @ |
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6517 |
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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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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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König v. Borstel, U.; Visser, E.K.; Hall, C. |
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Indicators of stress in equitation |
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Journal Article |
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2017 |
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Applied Animal Behaviour Science |
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Appl. Anim. Behav. Sci. |
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190 |
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43-56 |
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Stress; Horse; Riding; Heart rate variability; Cortisol; Behaviour |
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Abstract Stress is a generic concept describing the body's reaction to external stimuli, including both physiological and psychological factors. Therefore, by definition, the assessment of psychological stress in the exercising horse encompasses the problem of teasing apart the psychological and physiological factors both of which result in stress responses. The present study reviews the existing literature on various measures of stress taken specifically in the context of equitation science. Particular attention has been paid to short-term effects, and commonly used measurements of short-term stress include heart rate, a number of heart rate variability parameters, blood or saliva cortisol levels, eye temperature, and various behaviour parameters including in particular behaviour patterns presumably indicative of conflict with the rider's/trainer's aids. Inspection of the individual studies' results revealed that disagreement between these different measures of stress is commonplace. For physiological parameters, the largest proportion of agreement (i.e. both parameters simultaneously indicated either higher, insignificant or lower stress compared to a control treatment) was found for heart rate and heart rate variability parameters, while generally limited agreement was found for cortisol. It appears that cortisol levels may not be particularly useful for assessing/assessment of the valence of a situation in the exercising horse as cortisol levels are predominantly linked to activation and exercise levels. Although heart rate variability parameters reflect in theory more closely sympathovagal balance compared to cortisol levels, great care has to be taken regarding the use of appropriate time-frames, appropriate raw data correction methods as well as the use of appropriate equipment. In spite of its wide-spread and apparently successful use, popular equipment may in fact not be accurate enough under field conditions. Eye temperature is another promising parameter for assessment of psychological stress, but the technique is likewise susceptible to application errors. Given the high susceptibility of physiological parameters to errors at various experimental stages, behavioural rather than physiological parameters may in fact provide more accurate measures of valence when conducting experiments in the exercising horse. Behavioural parameters that appear to be particularly practical in assessing stress in ridden horses' behaviour are associated with frequencies of behaviour indicative of conflict. However, while increased frequencies of are a good indicator of stress, the absence of conflict behaviour does not provide proof of the absence of stress due to the possible occurrence of conditions such as Learned Helplessness. In future studies, the above issues should be taken into consideration when designing experiments to assess psychological stress in ridden horses. |
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0168-1591 |
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Equine Behaviour @ team @ |
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6160 |
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Aurich, J.; Wulf, M.; Ille, N.; Erber, R.; von Lewinski, M.; Palme, R.; Aurich, C. |
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Effects of season, age, sex and housing on salivary cortisol concentrations in horses |
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Journal Article |
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2015 |
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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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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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Year |
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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Wyss, C. |
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Equine Behaviour @ team @ |
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5869 |
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Nuñez, C.M.V.; Adelman, J.S.; Smith, J.; Gesquiere, L.R.; Rubenstein, D.I. |
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Linking social environment and stress physiology in feral mares (Equus caballus): Group transfers elevate fecal cortisol levels |
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Journal Article |
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2014 |
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General and Comparative Endocrinology |
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196 |
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26-33 |
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Equus caballus; Fecal cortisol; Feral mare; Group transfer; Stress; Social instability |
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Abstract Feral horses (Equus caballus) have a complex social structure, the stability of which is important to their overall health. Behavioral and demographic research has shown that decreases in group (or band) stability reduce female fitness, but the potential effects on the physiological stress response have not been demonstrated. To fully understand how band stability affects group-member fitness, we need to understand not only behavioral and demographic, but also physiological consequences of decreases to that stability. We studied group changes in feral mares (an activity that induces instability, including both male and female aggression) on Shackleford Banks, NC. We found that mares in the midst of changing groups exhibit increased fecal cortisol levels. In addition, mares making more group transfers show higher levels of cortisol two weeks post-behavior. These results offer insights into how social instability is integrated into an animal’s physiological phenotype. In addition, our results have important implications for feral horse management. On Shackleford Banks, mares contracepted with porcine zona pellucida (PZP) make approximately 10 times as many group changes as do untreated mares. Such animals may therefore be at higher risk of chronic stress. These results support the growing consensus that links between behavior and physiological stress must be taken into account when managing for healthy, functional populations. |
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0016-6480 |
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Equine Behaviour @ team @ |
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5743 |
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Kedzierski, W.; Wilk, I.; Janczarek, I. |
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Physiological response to the first saddling and first mounting of horses: comparison of two sympathetic training methods |
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Journal Article |
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2014 |
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Animal Science Papers and Reports |
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32 |
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3 |
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219-228 |
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cortisol / emotional reaction/ horses / natural training / stress |
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There is not much research done on the influence of sympathetic training on the emotional reaction
of horses. The aim of the present study was to evaluate the emotional response and the stress level
in horses to two sympathetic training methods: (1) with the use of the “round pen technique” (RP),
and (2) in which the RP was not applied (SH). Twenty two half-bred Anglo-Arab horses (2.5
years ±3 months of age) were subject to an initial training. Eleven horses were randomly included
to the RP method and the other 11 horses for the SH method. Heart rate (HR) and saliva cortisol
concentration were measured as indicators of horse emotional arousal and stress level, respectively.
The HR values were analysed: at rest, during the habituation period, just after the first saddling
and tightening of the girth, during the first time a human leaned over the horse’s back, and during
the mounting of the horse. Saliva samples were taken before and 15 min after each training session
studied. After saddling, the HR occurred significantly higher when the RP technique was used. The
significant increase in saliva cortisol concentration was observed only after the first mounting of
the horse. Generally, the use of the RP technique did not involve more important physiological
reactions in the trained horses than did the SH method. |
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Equine Behaviour @ team @ |
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5816 |
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Siniscalchi, M.; McFarlane, J.R.; Kauter, K.G.; Quaranta, A.; Rogers, L.J. |
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Title |
Cortisol levels in hair reflect behavioural reactivity of dogs to acoustic stimuli |
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Journal Article |
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Year |
2013 |
Publication |
Research in Veterinary Science |
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94 |
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1 |
Pages |
49-54 |
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Keywords |
Dogs; Behaviour; Cortisol; Hair; Acoustic stimuli |
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Abstract |
Cortisol levels in hair samples were examined in fourteen domestic dogs and related to the dogs’ responses to different acoustic stimuli. Stimuli were playbacks of species-typical vocalizations recorded during three different situations (“disturbance”, “isolation” and “play” barks) and the sounds of a thunderstorm. Hair samples were collected at 9:00 h and 17:00 h two weeks after the behavioural tests. Results showed that behavioural reactivity to playback of the various stimuli correlates with cortisol levels in hair samples collected at 9:00 h, and the same was the case for the separate measures of behaviour (i.e. hiding, running away, seeking attention from the tester, panting and lowering of the body posture). Hence, levels of cortisol in hair appear to reflect the dog’s chronic state of emotional reactivity, or temperament. |
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0034-5288 |
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Call Number |
Equine Behaviour @ team @ |
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5833 |
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