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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. | ||||
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| Notes | Approved | no | |||
| Call Number | Equine Behaviour @ team @ | Serial | 6520 | ||
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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 | ||||
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| Language | English | Summary Language | Original Title | ||
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| 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.; Moestl, E. | ||||
| Title | Measurement of cortisol metabolites in faeces of sheep as a parameter of cortisol concentration in blood | Type | Journal Article | ||
| Year | 1997 | Publication | Zeitschrift für Säugetierkunde | Abbreviated Journal | J. Mammal. Biol. |
| Volume | 62 | Issue | Pages | 192-197 | |
| Keywords | glucocorticoids, metabolites, animal biology, sheep, immunoenzyme techniques, | ||||
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| Notes | Approved | no | |||
| Call Number | Equine Behaviour @ team @ | Serial | 6044 | ||
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| Author |
Palme, R.; Fischer, P.; Schildorfer, H.; Ismail, M.N. | ||||
| Title | Excretion of infused 14C-steroid hormones via faeces and urine in domestic livestock | Type | 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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| Notes | Approved | no | |||
| Call Number | Equine Behaviour @ team @ | Serial | 4069 | ||
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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 | ||||
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| Language | English | Summary Language | Original Title | ||
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| ISSN | 0077-8923 | ISBN | Medium | ||
| Area | Expedition | Conference | |||
| Notes | PMID:16055844 | Approved | no | ||
| Call Number | Equine Behaviour @ team @ | Serial | 4081 | ||
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| Author |
Palme, R. | ||||
| Title | Monitoring stress hormone metabolites as a useful, non-invasive tool for welfare assessment in farm animals | Type | Journal Article | ||
| Year | 2012 | Publication | Animal Welfare | Abbreviated Journal | |
| Volume | 21 | Issue | 3 | Pages | 331-337 |
| Keywords | animal welfare, corticosterone, cortisol, faeces, farm animals, stress | ||||
| Abstract | A multitude of endocrine mechanisms are involved in coping with challenges. Glucocorticoids, secreted by the adrenal glands, are in the front line of the battle to overcome stressful situations. They are usually measured in plasma samples as parameters of adrenal activity and thus of disturbance. Unfortunately, collecting blood samples itself can disturb an animal. Thus, non-invasive methods for the determination of glucocorticoids or their metabolites have become increasingly popular. The pros and cons of various non-invasive sample materials (saliva, excreta, milk, hair/feathers and eggs) for glucocorticoid determination are given. Above all, faecal samples offer the advantage that they can be collected easily. In faecal samples, circulating hormone levels are integrated over a certain period of time and represent the cumulative secretion of hormones. Thus, the levels are less affected by short fluctuations or the pulse-like nature of hormone secretion. However, using this technique to assess an animal’s adrenocortical activity is not especially simple. Whether frequent sampling is necessary or single samples will suffice depends upon the study’s aim (whether one is examining the impact of acute or chronic stressors). Background knowledge of the metabolism and excretion of cortisol/corticosterone metabolites is required and a careful validation for each species and sex investigated is obligatory. The present review also addresses analytical issues regarding sample storage, extraction procedures and immunoassays and includes a comprehensive list of published studies (up to 2011) describing the use of such methods in farmed animals. Applied properly, non-invasive techniques to monitor glucocorticoid metabolites in faecal samples of various species are a useful tool for welfare assessment, especially as they are easily applied at farm or group level. |
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| ISSN | 0962-7286 | ISBN | Medium | ||
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| Notes | Approved | no | |||
| Call Number | Equine Behaviour @ team @ | Serial | 5793 | ||
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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 |
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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. |
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| 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. | ||||
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| 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 |
Möstl, E.; Palme, R. | ||||
| Title | Hormones as indicators of stress | Type | Journal Article | ||
| Year | 2002 | Publication | Domestic Animal Endocrinology | Abbreviated Journal | Domest. Anim. Endocrinol. |
| 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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| Series Volume | Series Issue | Edition | |||
| ISSN | 0739-7240 | ISBN | Medium | ||
| Area | Expedition | Conference | |||
| Notes | Approved | no | |||
| Call Number | Equine Behaviour @ team @ | Serial | 5930 | ||
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| 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 | ||||
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| Language | English | Summary Language | Original Title | ||
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| ISSN | 0077-8923 | ISBN | Medium | ||
| Area | Expedition | Conference | |||
| Notes | PMID:16055841 | Approved | no | ||
| Call Number | Equine Behaviour @ team @ | Serial | 4082 | ||
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