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Author |
Palme, R. |
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Title  |
Monitoring stress hormone metabolites as a useful, non-invasive tool for welfare assessment in farm animals |
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Journal Article |
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Year |
2012 |
Publication |
Animal Welfare |
Abbreviated Journal |
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Volume |
21 |
Issue |
3 |
Pages |
331-337 |
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Keywords |
animal welfare, corticosterone, cortisol, faeces, farm animals, stress |
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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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0962-7286 |
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Equine Behaviour @ team @ |
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5793 |
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Author |
Ganswindt, A.; Palme, R.; Heistermann, M.; Borragan, S.; Hodges, J.K. |
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Title |
Non-invasive assessment of adrenocortical function in the male African elephant (Loxodonta africana) and its relation to musth |
Type |
Journal Article |
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Year |
2003 |
Publication |
General and Comparative Endocrinology |
Abbreviated Journal |
Gen Comp Endocrinol |
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Volume |
134 |
Issue |
2 |
Pages |
156-166 |
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Keywords |
Adrenal Cortex/*metabolism/secretion; Adrenal Cortex Function Tests/methods/*veterinary; Adrenocorticotropic Hormone/physiology; Animals; Carbon Isotopes/diagnostic use; Chromatography, High Pressure Liquid/veterinary; Elephants/*metabolism/urine; Feces/*chemistry; Glucocorticoids/analysis/urine; Hydrocortisone/*analysis/diagnostic use/urine; Immunoenzyme Techniques/methods/veterinary; Male; Reproduction/physiology; Sexual Behavior, Animal/physiology; Stress, Psychological/diagnosis/*physiopathology; Testosterone/*analysis/diagnostic use/urine |
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Abstract |
Adult male elephants periodically show the phenomenon of musth, a condition associated with increased aggressiveness, restlessness, significant weight reduction and markedly elevated androgen levels. It has been suggested that musth-related behaviours are costly and that therefore musth may represent a form of physiological stress. In order to provide data on this largely unanswered question, the first aim of this study was to evaluate different assays for non-invasive assessment of adrenocortical function in the male African elephant by (i) characterizing the metabolism and excretion of [3H]cortisol (3H-C) and [14C]testosterone (14C-T) and (ii) using this information to evaluate the specificity of four antibodies for determination of excreted cortisol metabolites, particularly with respect to possible cross-reactions with androgen metabolites, and to assess their biological validity using an ACTH challenge test. Based on the methodology established, the second objective was to provide data on fecal cortisol metabolite concentrations in bulls during the musth and non-musth condition. 3H-C (1 mCi) and 14C-T (100 microCi) were injected simultaneously into a 16 year old male and all urine and feces collected for 30 and 86 h, respectively. The majority (82%) of cortisol metabolites was excreted into the urine, whereas testosterone metabolites were mainly (57%) excreted into the feces. Almost all radioactive metabolites recovered from urine were conjugated (86% 3H-C and 97% 14C-T). In contrast, 86% and >99% of the 3H-C and 14C-T metabolites recovered from feces consisted of unconjugated forms. HPLC separations indicated the presence of various metabolites of cortisol in both urine and feces, with cortisol being abundant in hydrolysed urine, but virtually absent in feces. Although all antibodies measured substantial amounts of immunoreactivity after HPLC separation of peak radioactive samples and detected an increase in glucocorticoid output following the ACTH challenge, only two (in feces against 3alpha,11-oxo-cortisol metabolites, measured by an 11-oxo-etiocholanolone-EIA and in urine against cortisol, measured by a cortisol-EIA) did not show substantial cross-reactivity with excreted 14C-T metabolites and could provide an acceptable degree of specificity for reliable assessment of glucocorticoid output from urine and feces. Based on these findings, concentrations of immunoreactive 3alpha,11-oxo-cortisol metabolites were determined in weekly fecal samples collected from four adult bulls over periods of 11-20 months to examine whether musth is associated with increased adrenal activity. Results showed that in each male levels of these cortisol metabolites were not elevated during periods of musth, suggesting that in the African elephant musth is generally not associated with marked elevations in glucocorticoid output. Given the complex nature of musth and the variety of factors that are likely to influence its manifestation, it is clear, however, that further studies, particularly on free-ranging animals, are needed before a possible relationship between musth and adrenal function can be resolved. This study also clearly illustrates the potential problems associated with cross-reacting metabolites of gonadal steroids in EIAs measuring glucocorticoid metabolites. This has to be taken into account when selecting assays and interpreting results of glucocorticoid metabolite analysis, not only for studies in the elephant but also in other species. |
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German Primate Centre, Department of Reproductive Biology, Kellnerweg 4, 37077 Gottingen, Germany. ganswindt@www.dpz.gdwg.de |
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English |
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0016-6480 |
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PMID:14511986 |
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Call Number |
Equine Behaviour @ team @ |
Serial |
4085 |
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Author |
Palme, R. |
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Title |
Non-invasive measurement of glucocorticoids: Advances and problems |
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Journal Article |
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Year |
2019 |
Publication |
Physiology & Behavior |
Abbreviated Journal |
Physiol. Behav. |
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Volume |
199 |
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Pages |
229-243 |
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Keywords |
Glucocorticoids; Non-invasive; Faecal cortisol/corticosterone metabolites; Immunoassays; Physiological/biological validation |
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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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0031-9384 |
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Palme2019_attachment.pdf |
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Call Number |
Equine Behaviour @ team @ |
Serial |
6517 |
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Author |
Palme, R. |
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Title |
Non-invasive monitoring of stress hormones for welfare assessment in domestic and wild equids |
Type |
Conference Article |
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Year |
2015 |
Publication |
Proceedings of the 3. International Equine Science Meeting |
Abbreviated Journal |
Proc. 3. Int. Equine. Sci. Mtg |
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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. |
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Xenophon Publishing |
Place of Publication |
Wald |
Editor |
Krueger, K. |
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Proc. 3. Int. Equine. Sci. Mtg |
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in prep |
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978-3-95625-000-2 |
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Id - |
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Equine Behaviour @ team @ |
Serial |
5795 |
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Author |
Merl, S.; Scherzer, S.; Palme, R.; Möstl, E. |
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Title |
Pain causes increased concentrations of glucocorticoid metabolites in horse feces |
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Journal Article |
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Year |
2000 |
Publication |
Journal of Equine Veterinary Science |
Abbreviated Journal |
J Equine Vet Sci |
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20 |
Issue |
9 |
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586-590 |
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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. |
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0737-0806 |
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no |
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Equine Behaviour @ team @ |
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6047 |
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Author |
Krueger, K.; Marr, I.; Dobler, A.; Palme, R. |
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Title |
Preservation of fecal glucocorticoid metabolites and immunoglobulin A through silica gel drying for field studies in horses |
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Journal Article |
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Year |
2019 |
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Conservation Physiology |
Abbreviated Journal |
conphys |
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7 |
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1 |
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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. |
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2051-1434 |
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Equine Behaviour @ team @ |
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6594 |
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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. |
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Title |
Rider effects on horses' conflict behaviour, rein tension, physiological measures and rideability scores |
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Journal Article |
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Year |
2021 |
Publication |
Applied Animal Behaviour Science |
Abbreviated Journal |
Appl. Anim. Behav. Sci. |
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234 |
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Pages |
105184 |
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Keywords |
Equitation science; Heart rate; Horse riding; Sport horse; Temperament; Training cues |
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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. |
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0168-1591 |
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Equine Behaviour @ team @ |
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6696 |
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Author |
Franceschini, C.; Siutz, C.; Palme, R.; Millesi, E. |
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Title |
Seasonal changes in cortisol and progesterone secretion in Common hamsters |
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Journal Article |
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Year |
2007 |
Publication |
General and Comparative Endocrinology |
Abbreviated Journal |
Gen Comp Endocrinol |
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152 |
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1 |
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14-21 |
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Animals; Cortisone/*secretion; Cricetinae/*metabolism; Feces/chemistry; Female; Lactation/physiology; Male; Pregnancy; Progesterone/*secretion; Reproduction/physiology; *Seasons; Sexual Behavior, Animal/physiology |
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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. |
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Address |
Department of Behavioural Biology, University of Vienna, Althanstrasse 14, A-1090 Vienna, Austria. claudia.franceschini@univie.ac.ct |
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0016-6480 |
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PMID:17408667 |
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Equine Behaviour @ team @ |
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4076 |
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Author |
Arlettaz, R.; Patthey, P.; Baltic, M.; Leu, T.; Schaub, M.; Palme, R.; Jenni-Eiermann, S. |
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Title |
Spreading free-riding snow sports represent a novel serious threat for wildlife |
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Journal Article |
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Year |
2007 |
Publication |
Proceedings. Biological Sciences / The Royal Society |
Abbreviated Journal |
Proc Biol Sci |
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274 |
Issue |
1614 |
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1219-1224 |
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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. |
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Address |
Zoological Institute, Division of Conservation Biology, University of Bern, Baltzerstrasse 6, 3012 Bern, Switzerland. raphael.arlettaz@nat.unibe.ch |
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English |
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0962-8452 |
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PMID:17341459 |
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no |
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Equine Behaviour @ team @ |
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4075 |
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Author |
Palme, R.; Touma, C.; Arias,N.; Dominchin, M.F.; Lepschy, M. |
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Title |
Steroid extraction: Get the best out of faecal samples |
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Journal Article |
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2012 |
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Veterinary Medicine Austria |
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Vet. Med. Austria |
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100 |
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238-246 |
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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. |
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Review, faeces, extrac- tion, non-invasive hormone moni- toring, stress, reproduction. |
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Equine Behaviour @ team @ |
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6046 |
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