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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	Palme, R.; Touma, C.; Arias,N.; Dominchin, M.F.; Lepschy, M.
	Title	Steroid extraction: Get the best out of faecal samples			Type	Journal Article
	Year	2012	Publication	Veterinary Medicine Austria	Abbreviated Journal	Vet. Med. Austria
	Volume	100	Issue		Pages	238-246
	Keywords
	Abstract	Faecal steroid hormone metabolites are becoming increasingly popular as parameters for reproductive functions and stress. Theextraction of the steroids from the faecal matrix represents the initial step before quantification can be performed. The steroid metabolites present in the faecal matrix are of varying polarity and composition, so selection of a proper extraction procedure is essential. There have been some studies to address this complex but often neglected point. Radiolabelled steroids (e.g. cortisol or progesterone) have frequently been added to faecal samples to estimate the efficiency of the extraction procedures used. However, native, unmetabolized steroids are normally not present in the faeces and therefore the results are artificial and do not accurately reflect the actual recoveries of the substances of interest. In this respect, recovery experiments based on faecal samples from radiometabolism studies are more informative. In these samples, the metabolite content accurately reflects the mixture of metabolites present in the given species. As a result, it is possible to evaluate different extraction methods for use with faecal samples. We present studies on sheep, horses, pigs, hares and dogs that utilized samples containing naturally metabolized, 14C-labelled steroids.
	Address	Review, faeces, extrac- tion, non-invasive hormone moni- toring, stress, reproduction.
	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		ISBN		Medium
	Area		Expedition		Conference
	Notes				Approved	no
	Call Number	Equine Behaviour @ team @			Serial	6046
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	Author	Palme, R.; Touma, C.; Arias, N., Dominchin, M.N.; Lepschy, M.
	Title	Steroid extraction: Get the best out of faecal samples			Type	Journal Article
	Year	2013	Publication	Wiener Tierärztliche Wochenschriften	Abbreviated Journal	Wien Tierärztl Monat – Vet Med Austria
	Volume	100	Issue		Pages	238-246.
	Keywords	Review, faeces, extraction, non-invasive hormone monitoring, stress, reproduction.
	Abstract	Faecal steroid hormone metabolites are becoming increasingly popular as parameters for reproductive functions and stress. The extraction of the steroids from the faecal matrix represents the initial step before quantification can be performed. The steroid metabolites present in the faecal matrix are of varying polarity and composition, so selection of a proper extraction procedure is essential. There have been some studies to address this complex but often neglected point. Radiolabelled steroids (e.g. cortisol or progesterone) have frequently been added to faecal samples to estimate the efficiency of the extraction procedures used. However, native, unmetabolized steroids are normally not present in the faeces and therefore the results are artifi- cial and do not accurately reflect the actual recoveries of the substances of interest. In this respect, recovery experiments based on faecal samples from radiometabolism studies are more informative. In these samples, the metabolite content accurately reflects the mixture of metabolites present in the given species. As a result, it is possible to evaluate different extraction methods for use with faecal samples. We present studies on sheep, horses, pigs, hares and dogs that utilized samples containing naturally metabolized, 14C-labelled steroids. We recommend extracting faecal steroids by simply suspending the faeces in a high percentage of a primary alcohol (for glucocorticoid metabolites 80% aqueous methanol proved best suited for virtually all mammalian species tested so far). Not only does the procedure significantly increase the total amount of recovered radioactivity, it also increases the percentage of unconjugated metabolites, which are more likely to be recognized by the antibodies used in various immunoassays. The advantages of this extraction procedure are clear: it is very easy to use (no evaporation step is needed), it yields high recoveries and variation based on the extraction procedure is reduced to a minimum.
	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		ISBN		Medium
	Area		Expedition		Conference
	Notes				Approved	no
	Call Number	Equine Behaviour @ team @			Serial	6520
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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	Franceschini, C.; Siutz, C.; Palme, R.; Millesi, E.
	Title	Seasonal changes in cortisol and progesterone secretion in Common hamsters			Type	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
	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	0016-6480	ISBN		Medium
	Area		Expedition		Conference
	Notes	PMID:17408667			Approved	no
	Call Number	Equine Behaviour @ team @			Serial	4076
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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	Krueger, K.; Marr, I.; Dobler, A.; Palme, R.
	Title	Preservation of fecal glucocorticoid metabolites and immunoglobulin A through silica gel drying for field studies in horses			Type	Journal Article
	Year	2019	Publication	Conservation Physiology	Abbreviated Journal	conphys
	Volume	7	Issue	1	Pages
	Keywords
	Abstract	Non-invasive methods enable stress evaluation through measuring fecal glucocorticoid metabolites (FGMs), and immunoglobulin A (IgA) in the feces avoiding stressful blood drawing or stressful restraining of animals in the field. However, FGMs and IgA are mostly analysed in freshly frozen samples, which is difficult when fresh samples cannot be frozen immediately or frozen samples cannot be stored or transported. Good results were also derived from air-dried fecal samples, which are hampered by unstable air humidity in the field. These difficulties may be overcome, when drying of samples could be induced with colorless silica gel (SiO2) granules in a secure set-up, such as an air tight tube. We determined the speed of drying 1.5 g of a fresh fecal sample from six horses on air and on silica gel. Furthermore, FGMs and IgA were analysed in differently stored subsamples from 12 horses: in frozen fecal samples, in air- or silica gel-dried samples stored for 1 day and for 7 days, and in wet fecal samples kept in a tube at room temperature for 7 days. FGM levels remained stable in feces dried on air or on silica gel for 7 days, whereas IgA quantities showed a significant loss. Under field conditions, when freezing or transporting the frozen samples is not possible and humidity hampers air drying, drying samples on silica gel in air tight tubes appears to be very helpful and reliable for analysing FGMs.
	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	2051-1434	ISBN		Medium
	Area		Expedition		Conference
	Notes				Approved	no
	Call Number	Equine Behaviour @ team @			Serial	6594
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	Author	Merl, S.; Scherzer, S.; Palme, R.; Möstl, E.
	Title	Pain causes increased concentrations of glucocorticoid metabolites in horse feces			Type	Journal Article
	Year	2000	Publication	Journal of Equine Veterinary Science	Abbreviated Journal	J Equine Vet Sci
	Volume	20	Issue	9	Pages	586-590
	Keywords
	Abstract	The concentration of 11,17-dioxoandrostanes (11,17-DOA), a group of cortisol metabolites, was measured using enzyme immunoassay in fecal samples of horses experiencing painful episodes. One group of horses consisted of 10 stallions castrated (samples were collected daily for 10 days); the other group was made up of 29 horses which were brought to an animal hospital because of signs of colic (samples were collected twice daily for six days). Before castration, median concentrations of 10.5 nmol/kg feces were measured. On days 1 and 2 after castration, median 11,17-DOA values increased up to 26.2 and 50.0 nmol/kg feces, respectively, and decreased thereafter to levels lower than at the beginning of the sampling period. High variations were measured between individual cases of colic. In animals with colic, all horses excreted more than 33 nmol 11,17-DOA/kg feces for various periods. The highest concentration measured was 885 nmol/kg feces. One animal out of the 29 colic horses did not show any clinical signs of pain upon arrival in the hospital. The 11,17-DOA values were below 17 nmol/kg feces in all those samples. From this data we conclude, that the concentration of 11,17-DOA in feces is a parameter for painful situations that have occurred one or two days earlier.
	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	0737-0806	ISBN		Medium
	Area		Expedition		Conference
	Notes				Approved	no
	Call Number	Equine Behaviour @ team @			Serial	6047
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	Author	Palme, R.
	Title	Non-invasive monitoring of stress hormones for welfare assessment in domestic and wild equids			Type	Conference Article
	Year	2015	Publication	Proceedings of the 3. International Equine Science Meeting	Abbreviated Journal	Proc. 3. Int. Equine. Sci. Mtg
	Volume		Issue		Pages
	Keywords
	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	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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