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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 |
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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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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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Year |
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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Anderson, M.K.; Friend, T.H.; Evans, J.W.; Bushong, D.M. |
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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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Dreschel, N.A.; Granger, D.A. |
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Title |
Methods of collection for salivary cortisol measurement in dogs |
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Journal Article |
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Year |
2009 |
Publication |
Hormones and Behavior |
Abbreviated Journal |
Horm. Behav. |
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55 |
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1 |
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163-168 |
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Dog; Canine; Salivary cortisol; Methods; Measurement; Stress |
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Salivary cortisol has been increasingly used as a measure of stress response in studies of welfare, reaction to stress and human–animal interactions in dogs and other species. While it can be a very useful measure, there are a number of saliva collection issues made evident through studies in the human and animal fields which have not been investigated in the canine species. Collection materials and the volume of saliva that is collected; the use of salivary stimulants; and the effect of food contamination can all dramatically impact cortisol measurement, leading to spurious results. In order to further examine the limitations of the collection method and the effects of collection material and salivary stimulant on salivary cortisol levels, a series of clinical, in vitro and in vivo studies were performed. It was found that there is a large amount of inter- and intra-individual variation in salivary cortisol measurement. Beef flavoring of collection materials leads to unpredictable variability in salivary cortisol concentration. Using salivary stimulants such as citric acid also has the potential to affect cortisol concentration measurement in saliva. Hydrocellulose appears to be a useful collection material for salivary cortisol determination. Recommendations for collection materials and use of salivary stimulants are presented. |
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0018-506x |
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Equine Behaviour @ team @ |
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5560 |
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Siniscalchi, M.; McFarlane, J.R.; Kauter, K.G.; Quaranta, A.; Rogers, L.J. |
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Cortisol levels in hair reflect behavioural reactivity of dogs to acoustic stimuli |
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Journal Article |
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2013 |
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Research in Veterinary Science |
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94 |
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1 |
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49-54 |
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Dogs; Behaviour; Cortisol; Hair; Acoustic stimuli |
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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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Equine Behaviour @ team @ |
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5833 |
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Ayala, I.; Martos, N.F.; Silvan, G.; Gutierrez-Panizo, C.; Clavel, J.G.; Illera, J.C. |
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Cortisol, adrenocorticotropic hormone, serotonin, adrenaline and noradrenaline serum concentrations in relation to disease and stress in the horse |
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2012 |
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Research in Veterinary Science |
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93 |
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1 |
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103-107 |
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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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Author |
Gröschl, M.; Wagner, R.; Rauh, M.; Dörr, H.G. |
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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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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 |
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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Young, T.; Creighton, E.; Smith, T.; Hosie, C. |
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A novel scale of behavioural indicators of stress for use with domestic horses |
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Journal Article |
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2012 |
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Applied Animal Behaviour Science |
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140 |
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1–2 |
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33-43 |
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Horse; Behaviour scores; Cortisol; Saliva; Welfare assessment; Non-invasive |
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Behaviour scores (BS) offer non-invasive, objective and easy to use ways of assessing welfare in animals. Their development has, however, largely focused on behavioural reactions to stressful events (often induced), and little use of physiological measures has been made to underpin and validate the behavioural measures. This study aimed to develop a physiologically validated scale of behavioural indicators of stress for the purpose of welfare assessment in stabled domestic horses. To achieve this, behavioural and physiological data were collected from 32 horses that underwent routine husbandry procedures. Principal component analysis (PCA) of the behavioural and physiological data revealed three meaningful components that were used as the basis of the scale. Analysis of video clips of the horses’ responses to the husbandry procedures was undertaken by a panel of equestrian industry professionals using a free choice profiling (FCP) methodology. These results were added to the scale along with key definitions from relevant literature. Salivary cortisol levels were significantly correlated with the BS confirming the scale was meaningful and reflected physiological stress. The scale offers an easy to use ‘tool’ for rapid, reliable non-invasive welfare assessment in horses, and reduces the need for potentially invasive physiological measures. |
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0168-1591 |
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Equine Behaviour @ team @ |
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5637 |
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Flauger, B.; Krueger, K.; Gerhards, H.; Möstl, E. |
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Simplified method to measure glucocorticoid metabolites in faeces of horses |
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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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