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Thiel, D., Jenni-Eiermann, S., & Palme, R. (2005). Measuring corticosterone metabolites in droppings of capercaillies (Tetrao urogallus). Ann N Y Acad Sci, 1046, 96–108.
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.
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
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Baltic, M., Jenni-Eiermann, S., Arlettaz, R., & Palme, R. (2005). A noninvasive technique to evaluate human-generated stress in the black grouse. Ann N Y Acad Sci, 1046, 81–95.
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.
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
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Palme, R. (2005). Measuring fecal steroids: guidelines for practical application. Ann N Y Acad Sci, 1046, 75–80.
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.
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Mostl, E., Rettenbacher, S., & Palme, R. (2005). Measurement of corticosterone metabolites in birds' droppings: an analytical approach. Ann N Y Acad Sci, 1046, 17–34.
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.
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Peel, J. A., Peel, M. B., & Davies, H. M. S. (2006). The effect of gallop training on hoof angle in thoroughbred racehorses. Equine Vet J Suppl, (36), 431–434.
Abstract: REASONS FOR PERFORMING STUDY: The economic impact of soundness problems in racehorses is very high and low hoof angle at the toe has been associated with a lack of soundness. However, it is not clear what environmental and management factors might contribute to a low hoof angle. OBJECTIVES: To investigate the hypothesis that the hooves of racehorses become flatter when in gallop training, as well as to determine factors contributing to this trend. METHODS: Weekly hoof measurements were taken with a hoof gauge from 45 Thoroughbred racehorses; 4 Thoroughbred show horses kept in consistent conditions and shod by the same farrier as some of the racehorses; and 6 unshod free-ranging horses. A further 15 horses were measured twice in one day to determine the repeatability of the method. RESULTS: Repeatability coefficients were 0.31 degrees for the left hoof and 0.37 degrees for the right. Racehorses in training showed a significant decrease in hoof angle over time while free ranging horses and show horses did not. Free-ranging horses had a significantly lower angle in winter (wet) compared with summer (dry) in both left (P = 0.040) and right (P = 0.017). Show horses had no significant change in hoof angle. Racehorses that had a period of rest during the experiment (n = 11) showed a decrease in hoof angle during training and an increase over their rest period for both hooves (P = 0.005 for the left hoof, P = 0.0009 for the right). CONCLUSIONS: Training for fast exercise in Thoroughbred racehorses is associated with a reduction in hoof angle and wet pasture conditions may also be associated with a reduced hoof angle in free-ranging horses. Potential relevance: Gallop exercise has a potentially large effect on hoof angle and therefore, a change in angle should be expected to occur in racehorses starting fast exercise work. Hence management of horses with abnormally low hoof angles may require an adaptation to their training regime in order to minimise this effect.
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Parsons, K. J., & Wilson, A. M. (2006). The use of MP3 recorders to log data from equine hoof mounted accelerometers. Equine Vet J, 38(7), 675–680.
Abstract: REASONS FOR PERFORMING STUDy: MP3 recorders are readily available, small, lightweight and low cost, providing the potential for logging analogue hoof mounted accelerometer signals for the characterisation of equine locomotion. These, however, require testing in practice. OBJECTIVES: To test whether 1) multiple MP3 recorders can maintain synchronisation, giving the ability to synchronise independent recorders for the logging of multiple limbs simultaneously; and 2) features of a foot mounted accelerometer signal attributable to foot-on and foot-off can be accurately identified from horse foot mounted accelerometers logged directly into an MP3 recorder. METHODS: Three experiments were performed: 1) Maintenance of synchronisation was assessed by counting the number of samples recorded by each of 4 MP3 recorders while mounted on a trotting horse and over 2 consecutive 30 min periods in 8 recorders on a bench. 2) Foot-on and foot-off times obtained from manual transcription of MP3 logged data and directly logged accelerometer signal were compared. 3) MP3/accelerometer acquisition units were used to log accelerometer signals from racehorses during extended training sessions. RESULTS: Mean absolute error of synchronisation between MP3 recorders was 10 samples per million (compared to mean number of samples, range 1-32 samples per million). Error accumulation showed a linear correlation with time. Features attributable to foot on and foot off were equally identifiable from the MP3 recorded signal over a range of equine gaits. CONCLUSIONS: Multiple MP3 recorders can be synchronised and used as a relatively cheap, robust, reliable and accurate logging system when combined with an accelerometer and external battery for the specific application of the measurement of stride timing variables across the range of equine gaits during field locomotion. POTENTIAL RELEVANCE: Footfall timings can be used to identify intervals between the fore and hind contacts, the identification of diagonal advanced placement and to calculate stride timing variables (stance time, protraction time and stride time). These parameters are invaluable for the characterisation and assessment of equine locomotion.
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Machnik, M., Hegger, I., Kietzmann, M., Thevis, M., Guddat, S., & Schanzer, W. (2007). Pharmacokinetics of altrenogest in horses. J Vet Pharmacol Ther, 30(1), 86–90.
Abstract: The Federation Equestre Internationale has permitted the use of altrenogest in mares for the control of oestrus. However, altrenogest is also suspicious to misuse in competition horses for its potential anabolic effects and suppression of typical male behaviour, and thus is a controlled drug. To investigate the pharmacokinetics of altrenogest in horses we conducted an elimination study. Five oral doses of 44 mug/kg altrenogest were administered to 10 horses at a dose interval of 24 h. Following administration blood and urine samples were collected at appropriate intervals. Altrenogest concentrations were measured by liquid chromatography-tandem mass spectrometry. The plasma levels of altrenogest reached maximal concentrations of 23-75 ng/mL. Baseline values were achieved within 3 days after the final administration. Urine peak concentrations of total altrenogest ranged from 823 to 3895 ng/mL. Twelve days after the final administration concentrations were below the limit of detection (ca 2 ng/mL).
Keywords: Administration, Oral; Animals; Chromatography, Liquid/veterinary; Doping in Sports/prevention & control; Horses/*metabolism; Male; Mass Spectrometry/veterinary; Progesterone Congeners/administration & dosage/blood/*pharmacokinetics/urine; Reproducibility of Results; Substance Abuse Detection/veterinary; Trenbolone/administration & dosage/*analogs & derivatives/blood/pharmacokinetics/urine
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Kralj-Fiser, S., Scheiber, I. B. R., Blejec, A., Moestl, E., & Kotrschal, K. (2007). Individualities in a flock of free-roaming greylag geese: behavioral and physiological consistency over time and across situations. Horm Behav, 51(2), 239–248.
Abstract: The concept of personality implies individual differences in behavior and physiology that show some degree of repeatability/consistency over time and across contexts. Most studies of animal personality, particularly studies of individuals' variation in physiological mechanisms, have been conducted on selected individuals in controlled conditions. We attempted to detect consistent behaviors as well as physiological patterns in greylag ganders (Anser anser) from a free-roaming flock living in semi-natural conditions. We tested 10 individuals repeatedly, in a handling trial, resembling tests for characterization of “temperaments” in captive animals. We recorded the behavior of the same 10 individuals during four situations in the socially intact flock: (1) a “low density feeding condition”, (2) a “high density feeding condition”, (3) a “low density post-feeding situation” and (4) while the geese rested. We collected fecal samples for determination of excreted immuno-reactive corticosterone (BM) and testosterone metabolites (TM) after handling trials, as well as the “low density feeding” and the “high density feeding” conditions. BM levels were very highly consistent over the repeats of handling trials, and the “low density feeding condition” and tended to be consistent over the first two repeats of the “high density feeding condition”. Also, BM responses tended to be consistent across contexts. Despite seasonal variation, there tended to be inter-test consistency of TM, which pointed to some individual differences in TM as well. Aggressiveness turned out to be a highly repeatable trait, which was consistent across social situations, and tended to correlate with an individual's resistance during handling trials. Also, “proximity to the female partner” and “sociability” – the average number of neighboring geese in a close distance while resting – were consistent. We conclude that aggressiveness, “affiliative tendencies” and levels of excreted corticosterone and testosterone metabolites may be crucial factors of personality in geese.
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