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Flauger, B.; Krueger, K.; Gerhards, H.; Möstl, E. |
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Title |
Simplified method to measure glucocorticoid metabolites in faeces of horses |
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Journal Article |
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2010 |
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Veterinary Research Communications |
Abbreviated Journal |
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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Equine Behaviour @ team @ |
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5073 |
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Gröschl, M.; Wagner, R.; Rauh, M.; Dörr, H.G. |
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Title |
Stability of salivary steroids: the influences of storage, food and dental care |
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2001 |
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Steroids |
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66 |
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10 |
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737-741 |
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Cortisol; 17OH-Progesterone; Progesterone; Saliva; Stability |
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We studied influences of dental care, food and storage on the reproducibility of salivary steroid levels. Cortisol (F), 17OH-progesterone (17OHP) and Progesterone (P) were measured using adapted commercial radioimmunoassays. Saliva samples of healthy adults (n = 15; m:8; f:7) were collected directly before and after dental care, and directly before and after breakfast with various foodstuffs. A second experiment investigated stability of steroids under different storage conditions. Four series of identical saliva portions (I: Native saliva; II: Centrifuged saliva; III: Saliva with trifluor acetate (TFA); IV: Saliva with 0.5% NaN3) were stored at room temperature and at 4°C for up to three weeks. To demonstrate influences of repeated thawing and re-freezing of saliva on steroid values, saliva samples (n = 15) were divided into identical portions. These portions were frozen and re-thawed up to 5 times before measurement. Neither dental care nor intake of bread or milk effected the reproducibility of F, 170HP, and P. Steroid levels decreased significantly in the course of three weeks under different storage conditions (P < 0.001). This decrease was clinically relevant from the second week onward, with exception of NaN3 treated samples. After repeated freezing and re-thawing 17OHP and P decreased slightly (about 5%). Only F decreased significantly after the third thawing (P < 0.001). The results show the usefulness of standardized handling of saliva samples for improving reproducibility and reliability of salivary steroid measurements. |
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0039-128x |
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Equine Behaviour @ team @ |
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5561 |
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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 |
Kedzierski, W.; Wilk, I.; Janczarek, I. |
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Title |
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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Year |
2014 |
Publication |
Animal Science Papers and Reports |
Abbreviated Journal |
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32 |
Issue |
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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König v. Borstel, U.; Visser, E.K.; Hall, C. |
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Title |
Indicators of stress in equitation |
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Journal Article |
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2017 |
Publication |
Applied Animal Behaviour Science |
Abbreviated Journal |
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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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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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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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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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 |
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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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Palme2019_attachment.pdf |
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Equine Behaviour @ team @ |
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6517 |
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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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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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Equine Behaviour @ team @ |
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5793 |
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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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2007 |
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Journal of Medical Primatology |
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36 |
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6 |
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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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Call Number |
Equine Behaviour @ team @ |
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5854 |
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