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	Author	Touma, C.; Palme, R.
	Title	Measuring fecal glucocorticoid metabolites in mammals and birds: the importance of validation			Type	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
	Corporate Author				Thesis
	Publisher		Place of Publication		Editor
	Language	English	Summary Language		Original Title
	Series Editor		Series Title		Abbreviated Series Title
	Series Volume		Series Issue		Edition
	ISSN	0077-8923	ISBN		Medium
	Area		Expedition		Conference
	Notes	PMID:16055843			Approved	no
	Call Number	Equine Behaviour @ team @			Serial	4073
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	Author	Thiel, D.; Jenni-Eiermann, S.; Palme, R.
	Title	Measuring corticosterone metabolites in droppings of capercaillies (Tetrao urogallus)			Type	Journal Article
	Year	2005	Publication	Annals of the New York Academy of Sciences	Abbreviated Journal	Ann N Y Acad Sci
	Volume	1046	Issue		Pages	96-108
	Keywords	Adrenocorticotropic Hormone/administration & dosage/analysis/metabolism; Animals; Circadian Rhythm; Corticosterone/administration & dosage/analysis/metabolism; Feces/chemistry; Female; Freezing; Galliformes/metabolism; Male; Reproducibility of Results; Sex Factors; Temperature; Time Factors; Tritium/diagnostic use
	Abstract	The capercaillie (Tetrao urogallus), the largest grouse species in the world, is decreasing in numbers in major parts of its distribution range. Disturbances by human outdoor activities are discussed as a possible reason for this population decline. An indicator for disturbances is the increase of the glucocorticoid corticosterone, a stress hormone, which helps to cope with life-threatening situations. However, repeated disturbances might result in a long-term increase of the basal corticosterone concentration, which can result in detrimental effects like reduced fitness and survival of an animal. To measure corticosterone metabolites (CMs) noninvasively in the droppings of free-living capercaillies, first an enzyme immunoassay (EIA) in captive birds had to be selected and validated. Therefore, the excretion pattern of intravenously injected radiolabeled corticosterone was determined and 3H metabolites were characterized. High-performance liquid chromatography (HPLC) separations of the samples containing peak concentrations revealed that corticosterone was extensively metabolized. The HPLC fractions were tested in several EIAs for glucocorticoid metabolites. The physiological relevance of this method was proved after pharmacological stimulation of the adrenocortical activity. Only the recently established cortisone assay, measuring CMs with a 3,11-dione structure, detected an expressed increase of concentrations following ACTH stimulation. To set up a sampling protocol suited for the field, we examined the influence of various storage conditions and time of day on concentrations of CMs.
	Address	Swiss Ornithological Institute, 6204 Sempach, Switzerland. dominik.thiel@vogelwarte.ch
	Corporate Author				Thesis
	Publisher		Place of Publication		Editor
	Language	English	Summary Language		Original Title
	Series Editor		Series Title		Abbreviated Series Title
	Series Volume		Series Issue		Edition
	ISSN	0077-8923	ISBN		Medium
	Area		Expedition		Conference
	Notes	PMID:16055846			Approved	no
	Call Number	Equine Behaviour @ team @			Serial	4079
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	Author	Baltic, M.; Jenni-Eiermann, S.; Arlettaz, R.; Palme, R.
	Title	A noninvasive technique to evaluate human-generated stress in the black grouse			Type	Journal Article
	Year	2005	Publication	Annals of the New York Academy of Sciences	Abbreviated Journal	Ann N Y Acad Sci
	Volume	1046	Issue		Pages	81-95
	Keywords	Adrenocorticotropic Hormone/metabolism; Animals; Bird Diseases/metabolism; Conservation of Natural Resources; Corticosterone/metabolism; Ecosystem; Feces/chemistry; Female; Galliformes/metabolism; Immunoenzyme Techniques/methods/veterinary; Male; Reproducibility of Results; Stress/metabolism/*veterinary; Tritium/diagnostic use
	Abstract	The continuous development of tourism and related leisure activities is exerting an increasingly intense pressure on wildlife. In this study, a novel noninvasive method for measuring stress in the black grouse, an endangered, emblematic species of European ecosystems that is currently declining in several parts of its European range, is tested and physiologically validated. A radiometabolism study and an ACTH challenge test were performed on four captive black grouse (two of each sex) in order to get basic information about the metabolism and excretion of corticosterone and to find an appropriate enzyme-immunoassay (EIA) to measure its metabolites in the feces. Peak radioactivity in the droppings was detected within 1 to 2 hours. Injected (3)H-corticosterone was excreted as polar metabolites and by itself was almost absent. A cortisone-EIA was chosen from among seven tested EIAs for different groups of glucocorticoid metabolites, because it cross-reacted with some of the formed metabolites and best reflected the increase of excreted corticosterone metabolites, after the ACTH challenge test. Concentrations of the metabolites from fecal samples collected from snow burrows of free-ranging black grouse were within the same range as in captive birds. The noninvasive method described may be appropriate for evaluating the stress faced by free-living black grouse populations in the wild, particularly in mountain ecosystems where human disturbance, especially by winter sports, is of increasing conservation concern.
	Address	Zoological Institute, Division of Conservation Biology, Baltzerstrasse 6, CH-3012 Bern, Switzerland
	Corporate Author				Thesis
	Publisher		Place of Publication		Editor
	Language	English	Summary Language		Original Title
	Series Editor		Series Title		Abbreviated Series Title
	Series Volume		Series Issue		Edition
	ISSN	0077-8923	ISBN		Medium
	Area		Expedition		Conference
	Notes	PMID:16055845			Approved	no
	Call Number	Equine Behaviour @ team @			Serial	4080
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	Author	Palme, R.
	Title	Measuring fecal steroids: guidelines for practical application			Type	Journal Article
	Year	2005	Publication	Annals of the New York Academy of Sciences	Abbreviated Journal	Ann N Y Acad Sci
	Volume	1046	Issue		Pages	75-80
	Keywords	Animals; Feces/chemistry; Immunoassay/methods; Reproducibility of Results; Specimen Handling/methods; Steroids/analysis
	Abstract	During the past 20 years, measuring steroid hormone metabolites in fecal samples has become a widely appreciated technique, because it has proved to be a powerful, noninvasive tool that provides important information about an animal's endocrine status (adrenocortical activity and reproductive status). However, although sampling is relatively easy to perform and free of feedback, a careful consideration of various factors is necessary to achieve proper results that lead to sound conclusions. This article aims to provide guidelines for an adequate application of these techniques. It is meant as a checklist that addresses the main topics of concern, such as sample collection and storage, time delay extraction procedures, assay selection and validation, biological relevance, and some confounding factors. These issues are discussed briefly here and in more detail in other recent articles.
	Address	Institute of Biochemistry, Department of Natural Sciences, University of Veterinary Medicine, Veterinaerplatz 1, A-1210 Vienna, Austria. Rupert.Palme@vu-wien.ac.at
	Corporate Author				Thesis
	Publisher		Place of Publication		Editor
	Language	English	Summary Language		Original Title
	Series Editor		Series Title		Abbreviated Series Title
	Series Volume		Series Issue		Edition
	ISSN	0077-8923	ISBN		Medium
	Area		Expedition		Conference
	Notes	PMID:16055844			Approved	no
	Call Number	Equine Behaviour @ team @			Serial	4081
Permanent link to this record



	Author	Mostl, E.; Rettenbacher, S.; Palme, R.
	Title	Measurement of corticosterone metabolites in birds' droppings: an analytical approach			Type	Journal Article
	Year	2005	Publication	Annals of the New York Academy of Sciences	Abbreviated Journal	Ann N Y Acad Sci
	Volume	1046	Issue		Pages	17-34
	Keywords	Animals; Birds/metabolism; Corticosterone/analysis/metabolism; Feces/*chemistry; Gas Chromatography-Mass Spectrometry; Immunoassay; Molecular Structure; Reproducibility of Results; Sensitivity and Specificity
	Abstract	Fecal steroid analyses are becoming increasingly popular among both field and laboratory scientists. The benefits associated with sampling procedures that do not require restraint, anesthesia, and blood collection include less risk to subject and investigator, as well as the potential to obtain endocrine profiles that are not influenced by the sampling procedure itself. In the feces, a species-specific pattern of metabolites is present, because glucocorticoids are extensively metabolized. Therefore, selection of adequate extraction procedures and immunoassays for measuring the relevant metabolites is a serious issue. In this review, emphasis is placed on the establishment and analytical validation of methods to measure glucocorticoid metabolites for a noninvasive evaluation of adrenocortical activity in droppings of birds.
	Address	Institute of Biochemistry, Department of Natural Sciences, University of Veterinary Medicine, Vienna, Veterinarplatz 1, A-1210 Vienna, Austria. erich.moestl@vu-wien.ac.at
	Corporate Author				Thesis
	Publisher		Place of Publication		Editor
	Language	English	Summary Language		Original Title
	Series Editor		Series Title		Abbreviated Series Title
	Series Volume		Series Issue		Edition
	ISSN	0077-8923	ISBN		Medium
	Area		Expedition		Conference
	Notes	PMID:16055841			Approved	no
	Call Number	Equine Behaviour @ team @			Serial	4082
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	Author	Palme, R.; Rettenbacher, S.; Touma, C.; El-Bahr, S.M.; Mostl, E.
	Title	Stress hormones in mammals and birds: comparative aspects regarding metabolism, excretion, and noninvasive measurement in fecal samples			Type	Journal Article
	Year	2005	Publication	Annals of the New York Academy of Sciences	Abbreviated Journal	Ann N Y Acad Sci
	Volume	1040	Issue		Pages	162-171
	Keywords	Adrenal Glands/chemistry/metabolism; Animals; Birds; Catecholamines/analysis/chemistry/metabolism; Feces/chemistry; Glucocorticoids/analysis/chemistry/metabolism; Hormones/analysis/metabolism; Mammals; Species Specificity; Stress/metabolism
	Abstract	A multitude of endocrine mechanisms are involved in coping with challenges. Front-line hormones to overcome stressful situations are glucocorticoids (GCs) and catecholamines (CAs). These hormones are usually determined in plasma samples as parameters of adrenal activity and thus of disturbance. GCs (and CAs) are extensively metabolized and excreted afterwards. Therefore, the concentration of GCs (or their metabolites) can be measured in various body fluids or excreta. Above all, fecal samples offer the advantages of easy collection and a feedback-free sampling procedure. However, large differences exist among species regarding the route and time course of excretion, as well as the types of metabolites formed. Based on information gained from radiometabolism studies (reviewed in this paper), we recently developed and successfully validated different enzyme immunoassays that enable the noninvasive measurement of groups of cortisol or corticosterone metabolites in animal feces. The determination of these metabolites in fecal samples can be used as a powerful tool to monitor GC production in various species of domestic, wildlife, and laboratory animals.
	Address	Institute of Biochemistry, Department of Natural Sciences, University of Veterinary Medicine, Vienna, Austria. rupert.palme@vu-wien.ac.at
	Corporate Author				Thesis
	Publisher		Place of Publication		Editor
	Language	English	Summary Language		Original Title
	Series Editor		Series Title		Abbreviated Series Title
	Series Volume		Series Issue		Edition
	ISSN	0077-8923	ISBN		Medium
	Area		Expedition		Conference
	Notes	PMID:15891021			Approved	no
	Call Number	Equine Behaviour @ team @			Serial	4083
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	Author	Arlettaz, R.; Patthey, P.; Baltic, M.; Leu, T.; Schaub, M.; Palme, R.; Jenni-Eiermann, S.
	Title	Spreading free-riding snow sports represent a novel serious threat for wildlife			Type	Journal Article
	Year	2007	Publication	Proceedings. Biological Sciences / The Royal Society	Abbreviated Journal	Proc Biol Sci
	Volume	274	Issue	1614	Pages	1219-1224
	Keywords
	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
	Corporate Author				Thesis
	Publisher		Place of Publication		Editor
	Language	English	Summary Language		Original Title
	Series Editor		Series Title		Abbreviated Series Title
	Series Volume		Series Issue		Edition
	ISSN	0962-8452	ISBN		Medium
	Area		Expedition		Conference
	Notes	PMID:17341459			Approved	no
	Call Number	Equine Behaviour @ team @			Serial	4075
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	Author	Christensen, J.W.; Munk, R.; Hawson, L.; Palme, R.; Larsen, T.; Egenvall, A.; König von Borstel, U.U.; Rørvang, M.V.
	Title	Rider effects on horses' conflict behaviour, rein tension, physiological measures and rideability scores			Type	Journal Article
	Year	2021	Publication	Applied Animal Behaviour Science	Abbreviated Journal	Appl. Anim. Behav. Sci.
	Volume	234	Issue		Pages	105184
	Keywords	Equitation science; Heart rate; Horse riding; Sport horse; Temperament; Training cues
	Abstract	Many breeding organisations include a subjective scoring of rideability by a professional rider into their evaluation of sports horses, but the consistency and reliability of the scoring system is debateable. The aim of this study was to investigate (i) whether professional riders agree in their scoring of rideability, and (ii) whether rideability scores are affected by rein tension, horse conflict behaviour, heart rate, and salivary cortisol, and (iii) whether riders induce different levels of conflict behaviour and physiological responses in the horses. Ten professional, female riders each rode 10 dressage horses (level M German scale; n = 100 combinations) through a standardised dressage test (10 min warm-up followed by a 4-min test) and subsequently scored the horses for rideability on the official 1-10 scale (1 = poor to 10 = excellent) from the Danish Riding Federation. Rein tension, horse heart rate, saliva cortisol and conflict behaviour were measured for each rider-horse pair. The riders were inconsistent in their scoring of rideability to the individual horses, e.g. scores for one of the horses ranged from 1 to 8. There was a significant effect of rider (P = 0.003) and the frequency of conflict behaviour (undesired head movements: P < 0.001, breaking the gait: P = 0.013, and other evasive behaviour: P = 0.032) on rideability scores, i.e. the more conflict behaviour the lower the score. There was no significant effect of rein tension and the physiological measures on rideability scores. However, there was a significant effect of rider on rein tension, horses' heart rate and increases in saliva cortisol concentrations and a tendency for some types of conflict behaviour, suggesting that some riders induced more discomfort in the horses. Future studies could help shed light on which elements of riding style are particularly important for sports horse welfare. In conclusion, this study found a large variation in rideability scores assigned to ten sports horses by ten professional riders. Rideability scores were dependent on the level of horse conflict behaviour, but not rein tension and physiological measures. Further studies are needed to improve the objectivity, consistency and reliability of rideability assessment of sports horses.
	Address
	Corporate Author				Thesis
	Publisher		Place of Publication		Editor
	Language		Summary Language		Original Title
	Series Editor		Series Title		Abbreviated Series Title
	Series Volume		Series Issue		Edition
	ISSN	0168-1591	ISBN		Medium
	Area		Expedition		Conference
	Notes				Approved	no
	Call Number	Equine Behaviour @ team @			Serial	6696
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	Author	Palme, R.
	Title	Non-invasive measurement of glucocorticoids: Advances and problems			Type	Journal Article
	Year	2019	Publication	Physiology & Behavior	Abbreviated Journal	Physiol. Behav.
	Volume	199	Issue		Pages	229-243
	Keywords	Glucocorticoids; Non-invasive; Faecal cortisol/corticosterone metabolites; Immunoassays; Physiological/biological validation
	Abstract	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.
	Address
	Corporate Author				Thesis
	Publisher		Place of Publication		Editor
	Language		Summary Language		Original Title
	Series Editor		Series Title		Abbreviated Series Title
	Series Volume		Series Issue		Edition
	ISSN	0031-9384	ISBN		Medium
	Area		Expedition		Conference
	Notes	Palme2019_attachment.pdf			Approved	no
	Call Number	Equine Behaviour @ team @			Serial	6517
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	Author	Sheriff, M.J.; Dantzer, B.; Delehanty, B.; Palme, R.; Boonstra, R.
	Title	Measuring stress in wildlife: techniques for quantifying glucocorticoids			Type	Journal Article
	Year	2011	Publication	Oecologia	Abbreviated Journal
	Volume	166	Issue	4	Pages	869-887
	Keywords
	Abstract	Stress responses play a key role in allowing animals to cope with change and challenge in the face of both environmental certainty and uncertainty. Measurement of glucocorticoid levels, key elements in the neuroendocrine stress axis, can give insight into an animal’s well-being and can aid understanding ecological and evolutionary processes as well as conservation and management issues. We give an overview of the four main biological samples that have been utilized [blood, saliva, excreta (feces and urine), and integumentary structures (hair and feathers)], their advantages and disadvantages for use with wildlife, and some of the background and pitfalls that users must consider in interpreting their results. The matrix of choice will depend on the nature of the study and of the species, on whether one is examining the impact of acute versus chronic stressors, and on the degree of invasiveness that is possible or desirable. In some cases, more than one matrix can be measured to achieve the same ends. All require a significant degree of expertise, sometimes in obtaining the sample and always in extracting and analyzing the glucocorticoid or its metabolites. Glucocorticoid measurement is proving to be a powerful integrator of environmental stressors and of an animal’s condition.
	Address
	Corporate Author				Thesis
	Publisher		Place of Publication		Editor
	Language		Summary Language		Original Title
	Series Editor		Series Title		Abbreviated Series Title
	Series Volume		Series Issue		Edition
	ISSN	1432-1939	ISBN		Medium
	Area		Expedition		Conference
	Notes				Approved	no
	Call Number	Equine Behaviour @ team @ Sheriff2011			Serial	6150
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