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Author Christensen, J.W.; Ahrendt, L.P.; Lintrup, R.; Gaillard, C.; Palme, R.; Malmkvist, J. doi  openurl
  Title Does learning performance in horses relate to fearfulness, baseline stress hormone, and social rank? Type (up) Abstract
  Year 2012 Publication Applied Animal Behaviour Science Abbreviated Journal App Anim Behav Sci  
  Volume 140 Issue 1 Pages 44-52  
  Keywords Horse; Learning; Fearfulness; Stress; Reinforcement; Social rank  
  Abstract The ability of horses to learn and remember new tasks is fundamentally important for their use by humans. Fearfulness may, however, interfere with learning, because stimuli in the environment can overshadow signals from the rider or handler. In addition, prolonged high levels of stress hormones can affect neurons within the hippocampus; a brain region central to learning and memory. In a series of experiments, we aimed to investigate the link between performance in two learning tests, the baseline level of stress hormones, measured as faecal cortisol metabolites (FCM), fearfulness, and social rank. Twenty-five geldings (2 or 3 years old) pastured in one group were included in the study. The learning tests were performed by professional trainers and included a number of predefined stages during which the horses were gradually trained to perform exercises, using either negative (NR) or positive reinforcement (PR). Each of the learning tests lasted 3 days; 7min/horse/day. The NR test was repeated in a novel environment. Performance, measured as final stage in the training programme, and heart rate (HR) were recorded. Faeces were collected on four separate days where the horses had been undisturbed at pasture for 48h. Social rank was determined through observations of social interactions during feeding. The fear test was a novel object test during which behaviour and HR were recorded. Performance in the NR and PR learning tests did not correlate. In the NR test, there was a significant, negative correlation between performance and HR in the novel environment (rS=-0.66, P<0.001, i.e. nervous horses had reduced performance), whereas there was no such correlation in the home environment (both NR and PR). Behavioural reactions in the fear test correlated significantly with performance in the NR test in the novel environment (e.g. object alertness and final stage: rS=-0.43, P=0.04), suggesting that performance under unfamiliar, stressful conditions may be predicted by behavioural responses in a fear test. There was a negative correlation between social rank and baseline stress hormones (rS=-0.43, P=0.04), i.e. high rank corresponded to low FCM concentrations, whereas neither rank nor FCM correlated with fearfulness or learning performance. We conclude that performance under stressful conditions is affected by activation of the sympathetic nervous system during training and related to behavioural responses in a standardised fear test. Learning performance in the home environment, however, appears unrelated to fearfulness, social rank and baseline FCM levels.  
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  ISSN 0168-1591 ISBN Medium  
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  Notes Approved no  
  Call Number Equine Behaviour @ team @ S0168-1591(12)00168-2 Serial 5769  
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Author Palme, R. pdf  isbn
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  Title Non-invasive monitoring of stress hormones for welfare assessment in domestic and wild equids Type (up) Conference Article
  Year 2015 Publication Proceedings of the 3. International Equine Science Meeting Abbreviated Journal Proc. 3. Int. Equine. Sci. Mtg  
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  Abstract Stress responses play an important role in allowing animals to cope with challenges. Glucocorticoids, key elements in the neuroendocrine stress axis, are traditionally measured as a parameter for welfare assessment. As blood sample collection itself disturbs an animal, non-invasive or minimal invasive methods have gained importance for assessing stress. In horses saliva and faeces are most frequently used. Faecal samples offer the advantage that they can be collected easily and stress-free. In faecal samples circulating hormone levels are integrated over a certain period of time. As a consequence faecal glucocorticoid metabolites represent the cumulative secretion and they are less affected by short episodic fluctuations of hormone secretion.

However, in order to gain reliable information about an animal’s adrenocortical activity, certain criteria have to be met: Depending whether the impact of acute or chronic stressors is assessed, frequent sampling might be necessary whereas in other cases, single samples will suffice. Background knowledge regarding the metabolism and excretion of glucocorticoids is essential and a careful validation is obligatory. In addition, this presentation will address analytical issues regarding sample storage, extraction procedures, and immunoassays and various examples of a successful application in equids will be given. Applied properly, non-invasive techniques to monitor stress hormones are a useful tool for animal welfare assessment.
 
  Address  
  Corporate Author Palme, R. Thesis  
  Publisher Xenophon Publishing Place of Publication Wald Editor Krueger, K.  
  Language Summary Language Original Title  
  Series Editor Series Title Abbreviated Series Title Proc. 3. Int. Equine. Sci. Mtg  
  Series Volume in prep Series Issue Edition  
  ISSN ISBN 978-3-95625-000-2 Medium  
  Area Expedition Conference  
  Notes Id - Approved no  
  Call Number Equine Behaviour @ team @ Serial 5795  
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Author Schwarzenberger, F.; Mostl, E.; Palme, R.; Bamberg, E. url  openurl
  Title Faecal steroid analysis for non-invasive monitoring of reproductive status in farm, wild and zoo animals Type (up) Journal Article
  Year 1996 Publication Animal Reproduction Science Abbreviated Journal Animal Reproduction: Research and Practice  
  Volume 42 Issue 1-4 Pages 515-526  
  Keywords Faecal steroids; Non-invasive monitoring; Oestrogens; Progesterone metabolites; Reproductive hormones  
  Abstract Non-invasive faecal oestrogen and progesterone metabolite evaluations are well established approaches for monitoring reproductive function in a variety of mammalian species. The route of excretion of steroid hormone metabolites varies considerably among species, and also between steroids within the same species. Steroid concentrations in faeces exhibit a similar pattern to those in plasma, but have a lag time, which depending upon the species, can be from 12 h to more than 2 days. Faecal steroid metabolites in mammals are mainly unconjugated compounds. Faecal oestrogens consist predominantly of oestrone and/or oestradiol-17α or -17β. Therefore, specific oestrogen antibodies or antibodies against total oestrogens can be used for their determination. Progesterone is metabolised to several 5α- or 5β-reduced pregnanediones and hydroxylated pregnanes prior to its faecal excretion. Therefore, relevant antibodies for their determination show considerable cross-reactivities with several pregnane metabolites, whereas specific progesterone antibodies are less suitable. Faecal oestrogen evaluations have been used as reliable indicators of pregnancy in several ungulate and some primate species. They have also been used to determine the preovulatory period in carnivores, corpus luteum activity in New World primates, and to diagnose cryptorchidism in horses. Faecal progesterone metabolite analysis has been successfully used for monitoring corpus luteum function and pregnancy, abortion, seasonality and treatment therapies in an ever expanding list of species.  
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  Call Number refbase @ user @ Serial 327  
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Author Mostl, E.; Palme, R. url  openurl
  Title Hormones as indicators of stress Type (up) Journal Article
  Year 2002 Publication Domestic Animal Endocrinology Abbreviated Journal Fourth International Conference on Farm Animal Endocrinology  
  Volume 23 Issue 1-2 Pages 67-74  
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  Abstract Animal welfare is of increasing importance and absence of chronic stress is one of its prerequisites. During stress, various endocrine responses are involved to improve the fitness of the individual. The front-line hormones to overcome stressful situations are the glucocorticoids and catecholamines. These hormones are determined as a parameter of adrenal activity and thus of disturbance. The concentration of glucocorticoids (or their metabolites) can be measured in various body fluids or excreta. Above all, fecal samples offer the advantage that they can be easily collected and this procedure is feedback free. Recently, enzyme immunoassays (EIA) have been developed and successfully tested, to enable the measurement of groups of cortisol metabolites in animal feces. The determination of these metabolites in fecal samples is a practical method to monitor glucocorticoid production.  
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  Notes Approved no  
  Call Number Equine Behaviour @ team @ Serial 4067  
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Author Palme, R.; Fischer, P.; Schildorfer, H.; Ismail, M.N. url  openurl
  Title Excretion of infused 14C-steroid hormones via faeces and urine in domestic livestock Type (up) Journal Article
  Year 1996 Publication Animal Reproduction Science Abbreviated Journal  
  Volume 43 Issue 1 Pages 43-63  
  Keywords Sheep--endocrinology; Pig--endocrinology; Pony; 14C-steroids; Faeces; Urine; Blood  
  Abstract The aim of this comparative study was to gain more information about the excretion of steroid hormones in farm animals. This should help to establish or improve non-invasive steroid monitoring procedures, especially in zoo and wildlife animals. Over a period of 4 h the 14C-steroid hormones (3.7 MBq) progesterone (three females), testosterone (three males), cortisol and oestrone (two males, two females) were infused intravenously in sheep, ponies and pigs. Faeces were collected immediately after defecation. Urine was sampled via a permanent catheter in females and after spontaneous urination in males. A total of 88 +/- 10% (mean +/- SD) of the administered radioactivity was recovered. Considerable interspecies differences were measured both in the amounts of steroid metabolites excreted via faeces or urine and the time course of excretion. Progesterone and oestrone in ewes, and progesterone in mares were excreted mainly in the faeces (over 75%). The primary route of excretion of all other 14C-steroids was via the urine but to a different extent. In general, sheep showed the highest degree of faecal excretion and pigs the least. The highest radioactivity in urine (per mmol creatinine) was observed during the infusion or in one of the next two samples thereafter, whereas in faeces it was measured about 12 h (sheep), 24 h (ponies) or 48 h (pigs) after the end of the infusion. Thereafter the radioactivity declined and reached background levels within 2-3 weeks. In faeces, steroid metabolites were present mainly in an unconjugated form, but in blood and urine as conjugates. Mean retention time of faecal radioactivity suggested that the passage rate of digesta (duodenum to rectum) played an important role in the time course of the excretion of steroids. The information derived from this investigation could improve the precision of sampling as well as the extraction of steroids from the faeces. Furthermore, the study demonstrates that it should be possible to establish methods for measuring faecal androgen and cortisol metabolites for assessing male reproductive endocrinology and stress in animals.  
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  Call Number Equine Behaviour @ team @ Serial 4069  
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Author Touma, C.; Palme, R. doi  openurl
  Title Measuring fecal glucocorticoid metabolites in mammals and birds: the importance of validation Type (up) Journal Article
  Year 2005 Publication Annals of the New York Academy of Sciences Abbreviated Journal Ann N Y Acad Sci  
  Volume 1046 Issue Pages 54-74  
  Keywords Animals; Birds/*metabolism; Circadian Rhythm; Feces/*chemistry; Glucocorticoids/*analysis; Mammals/*metabolism; Reproducibility of Results; Seasons; Sex Factors  
  Abstract In recent years, the noninvasive monitoring of steroid hormone metabolites in feces of mammals and droppings of birds has become an increasingly popular technique. It offers several advantages and has been applied to a variety of species under various settings. However, using this technique to reliably assess an animal's adrenocortical activity is not that simple and straightforward to apply. Because clear differences regarding the metabolism and excretion of glucocorticoid metabolites (GCMs) exist, a careful validation for each species and sex investigated is obligatory. In this review, general analytical issues regarding sample storage, extraction procedures, and immunoassays are briefly discussed, but the main focus lies on experiments and recommendations addressing the validation of fecal GCM measurements in mammals and birds. The crucial importance of scrutinizing the physiological and biological validity of fecal GCM analyses in a given species is stressed. In particular, the relevance of the technique to detect biologically meaningful alterations in adrenocortical activity must be shown. Furthermore, significant effects of the animals' sex, the time of day, season, and different life history stages are discussed, bringing about the necessity to seriously consider possible sex differences as well as diurnal and seasonal variations. Thus, comprehensive information on the animals' biology and stress physiology should be carefully taken into account. Together with an extensive physiological and biological validation, this will ensure that the measurement of fecal GCMs can be used as a powerful tool to assess adrenocortical activity in diverse investigations on laboratory, companion, farm, zoo, and wild animals.  
  Address Max Planck Institute of Psychiatry, Department of Behavioral Neuroendocrinology, Kraepelinstrasse 2-10, D-80804 Munich, Germany. touma@mpipsykl.mpg.de  
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  ISSN 0077-8923 ISBN Medium  
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  Notes PMID:16055843 Approved no  
  Call Number Equine Behaviour @ team @ Serial 4073  
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Author Arlettaz, R.; Patthey, P.; Baltic, M.; Leu, T.; Schaub, M.; Palme, R.; Jenni-Eiermann, S. doi  openurl
  Title Spreading free-riding snow sports represent a novel serious threat for wildlife Type (up) Journal Article
  Year 2007 Publication Proceedings. Biological Sciences / The Royal Society Abbreviated Journal Proc Biol Sci  
  Volume 274 Issue 1614 Pages 1219-1224  
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  Abstract Stress generated by humans on wildlife by continuous development of outdoor recreational activities is of increasing concern for biodiversity conservation. Human disturbance often adds to other negative impact factors affecting the dynamics of vulnerable populations. It is not known to which extent the rapidly spreading free-riding snow sports actually elicit detrimental stress (allostatic overload) upon wildlife, nor what the potential associated fitness and survival costs are. Using a non-invasive technique, we evaluated the physiological stress response induced by free-riding snow sports on a declining bird species of Alpine ecosystems. The results of a field experiment in which radiomonitored black grouse (Tetrao tetrix) were actively flushed from their snow burrows once a day during four consecutive days showed an increase in the concentration of faecal stress hormone (corticosterone) metabolites after disturbance. A large-scale comparative analysis across the southwestern Swiss Alps indicated that birds had higher levels of these metabolites in human-disturbed versus undisturbed habitats. Disturbance by snow sport free-riders appears to elevate stress, which potentially represents a new serious threat for wildlife. The fitness and survival costs of allostatic adjustments have yet to be estimated.  
  Address Zoological Institute, Division of Conservation Biology, University of Bern, Baltzerstrasse 6, 3012 Bern, Switzerland. raphael.arlettaz@nat.unibe.ch  
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  Series Volume Series Issue Edition  
  ISSN 0962-8452 ISBN Medium  
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  Notes PMID:17341459 Approved no  
  Call Number Equine Behaviour @ team @ Serial 4075  
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Author Franceschini, C.; Siutz, C.; Palme, R.; Millesi, E. doi  openurl
  Title Seasonal changes in cortisol and progesterone secretion in Common hamsters Type (up) Journal Article
  Year 2007 Publication General and Comparative Endocrinology Abbreviated Journal Gen Comp Endocrinol  
  Volume 152 Issue 1 Pages 14-21  
  Keywords Animals; Cortisone/*secretion; Cricetinae/*metabolism; Feces/chemistry; Female; Lactation/physiology; Male; Pregnancy; Progesterone/*secretion; Reproduction/physiology; *Seasons; Sexual Behavior, Animal/physiology  
  Abstract In this study, we investigated endocrine factors and behaviour in free-living Common hamsters (Cricetus cricetus) during reproductive and non-reproductive periods of the annual cycle. We applied a non-invasive method to gain information on seasonal changes in adrenocortical activity in male and female hamsters by analysing faecal glucocorticoid metabolite concentrations (FCM). In addition, plasma progesterone concentrations were monitored in females throughout the non-hibernation season. The animals were live-trapped from spring emergence until the onset of hibernation in autumn. Reproductive status was determined at capture and blood and faecal samples were collected. During behavioural observations, agonistic and sexual interactions were recorded. FCM concentrations were significantly higher in males than in females during the reproductive period. In males, a pronounced increase in FCM during the reproductive period coincided with high frequencies of intrasexual aggression. In females, FCM levels remained relatively constant. Aggressive behaviour in females increased during the reproductive period, but was much less frequent than in males. Females, which successfully raised a second litter after a postpartum oestrus and concurrent lactation and gestation had lower FCM levels than individuals, which lost their second litter after parturition. As expected, plasma progesterone concentrations were low before and after the reproductive period. During gestation, levels peaked and remained elevated during lactation. The results of this field study provide insight in critical periods associated with reproduction in male and female Common hamsters.  
  Address Department of Behavioural Biology, University of Vienna, Althanstrasse 14, A-1090 Vienna, Austria. claudia.franceschini@univie.ac.ct  
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  ISSN 0016-6480 ISBN Medium  
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  Notes PMID:17408667 Approved no  
  Call Number Equine Behaviour @ team @ Serial 4076  
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Author Szenci, O.; Palme, R.; Taverne, M.A.; Varga, J.; Meersma, N.; Wissink, E. openurl 
  Title Evaluation of false ultrasonographic pregnancy diagnoses in sows by measuring the concentration of unconjugated estrogens in feces Type (up) Journal Article
  Year 1997 Publication Theriogenology Abbreviated Journal Theriogenology  
  Volume 48 Issue 5 Pages 873-882  
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  Abstract On Days 26, 28, and 30 after AI, ultrasonographic pregnancy diagnoses were performed on 207 gilts and sows by using a 3.5 MHz linear-array transducer. Fecal samples were taken from the rectum after each ultrasonographic examination, and the concentrations of unconjugated estrogens in selected samples (n = 73) were measured by RIA. Fecal unconjugated estrogen concentration of 11.7 ng/g feces or higher was indicative of pregnancy. The overall sensitivity and specificity of the ultrasonographic test was 99% for farrowing sows and 73.1% for nonfarrowing sows. With one exception, sows with a false negative diagnosis by ultrasonography on Day 26 were correctly diagnosed pregnant by elevated fecal unconjugated estrogens or repeated ultrasonographic examinations on Days 28 or 30. Return to estrus around the sampling period may cause false positive results in the unconjugated estrogen assay, while early embryonic mortality can result in false positive diagnoses in both the ultrasonographic test and estrogen assay. Although there was a positive correlation between the concentrations of unconjugated estrogens in the feces and litter size at farrowing in the selected sows, it seems very unlikely that fecal estrogens can provide an accurate tool for predicting litter size.  
  Address University of Veterinary Science, Department of Obstetrics and Reproduction H-1400 Budapest, P.O. Box 2, Hungary  
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  ISSN 0093-691X ISBN Medium  
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  Notes PMID:16728179 Approved no  
  Call Number Equine Behaviour @ team @ Serial 4077  
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Author Heistermann, M.; Palme, R.; Ganswindt, A. doi  openurl
  Title Comparison of different enzyme-immunoassays for assessment of adrenocortical activity in primates based on fecal analysis Type (up) Journal Article
  Year 2006 Publication American journal of primatology Abbreviated Journal Am. J. Primatol.  
  Volume 68 Issue 3 Pages 257-273  
  Keywords 11-Hydroxycorticosteroids/*analysis; Adrenocorticotropic Hormone/pharmacology; Anesthesia; Animals; Corticosterone/analysis; Feces/*chemistry; Glucocorticoids/*analysis; Haplorhini/*metabolism; Hydrocortisone/analysis; Hypothalamo-Hypophyseal System/drug effects/physiology; Immunoenzyme Techniques/*methods; Pituitary-Adrenal System/drug effects/physiology; Species Specificity  
  Abstract Most studies published to date that used fecal glucocorticoid measurements to assess adrenocortical activity in primate (and many nonprimate) species applied a specific cortisol or corticosterone assay. However, since these native glucocorticoids are virtually absent in the feces of most vertebrates, including primates, the validity of this approach has recently been questioned. Therefore, the overall aim of the present study was to assess the validity of four enzyme-immunoassays (EIAs) using antibodies raised against cortisol, corticosterone, and reduced cortisol metabolites (two group-specific antibodies) for assessing adrenocortical activity using fecal glucocorticoid metabolite (GCM) measurements in selected primate species (marmoset, long-tailed macaque, Barbary macaque, chimpanzee, and gorilla). Using physiological stimulation of the hypothalamo-pituitary-adrenocortical (HPA) axis by administering exogenous ACTH or anesthesia, we demonstrated that at least two assays detected the predicted increase in fecal GCM levels in response to treatment in each species. However, the magnitude of response varied between assays and species, and no one assay was applicable to all species. While the corticosterone assay generally was of only limited suitability for assessing glucocorticoid output, the specific cortisol assay was valuable for those species that (according to high-performance liquid chromatography (HPLC) analysis data) excreted clearly detectable amounts of authentic cortisol into the feces. In contrast, in species in which cortisol was virtually absent in the feces, group-specific assays provided a much stronger signal, and these assays also performed well in the other primate species tested (except the marmoset). Collectively, the data suggest that the reliability of a given fecal glucocorticoid assay in reflecting activity of the HPA axis in primates clearly depends on the species in question. Although to date there is no single assay system that can be used successfully across species, our data suggest that group-specific assays have a high potential for cross-species application. Nevertheless, regardless of which GC antibody is chosen, our study clearly reinforces the necessity of appropriately validating the respective assay system before it is used.  
  Address Department of Reproductive Biology, German Primate Center, Gottingen, Germany. mheiste@gwdg.de  
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  ISSN 0275-2565 ISBN Medium  
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  Notes PMID:16477600 Approved no  
  Call Number Equine Behaviour @ team @ Serial 4078  
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