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Touma, C., Palme, R., & Sachser, N. (2004). Analyzing corticosterone metabolites in fecal samples of mice: a noninvasive technique to monitor stress hormones. Horm Behav, 45(1), 10–22.
Abstract: In small animals like mice, the monitoring of endocrine functions over time is constrained seriously by the adverse effects of blood sampling. Therefore, noninvasive techniques to monitor, for example, stress hormones in these animals are highly demanded in laboratory as well as in field research. The aim of our study was to evaluate the biological relevance of a recently developed technique to monitor stress hormone metabolites in fecal samples of laboratory mice. In total, six experiments were performed using six male and six female mice each. Two adrenocorticotropic hormone (ACTH) challenge tests, two dexamethasone (Dex) suppression tests and two control experiments [investigating effects of the injection procedure itself and the diurnal variation (DV) of glucocorticoids (GCs), respectively] were conducted. The experiments clearly demonstrated that pharmacological stimulation and suppression of adrenocortical activity was reflected accurately by means of corticosterone metabolite (CM) measurements in the feces of males and females. Furthermore, the technique proved sensitive enough to detect dosage-dependent effects of the ACTH/Dex treatment and facilitated to reveal profound effects of the injection procedure itself. Even the naturally occurring DV of GCs could be monitored reliably. Thus, our results confirm that measurement of fecal CM with the recently established 5alpha-pregnane-3beta,11beta,21-triol-20-one enzyme immunoassay is a very powerful tool to monitor adrenocortical activity in laboratory mice. Since mice represent the vast majority of all rodents used for research worldwide and the number of transgenic and knockout mice utilized as animal models is still increasing, this noninvasive technique can open new perspectives in biomedical and behavioral science.
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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.
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Stahl, F., & Dorner, G. (1982). Responses of salivary cortisol levels to stress-situations. Endokrinologie, 80(2), 158–162.
Abstract: A procedure is described for determining salivary cortisol levels by a competitive protein-binding assay using horse transcortin. The collection of saliva was performed by means of filter paper-strips. Filter paper samples are more than 5 days stable after air-drying. In this form, the samples could be stored without refrigerator or deep-freezer and, if necessary, sent by post to the laboratory without any special precaution. Stressful situation of either painful or anxious origin were associated with an adequate increase of salivary cortisol levels. The increases were 157 to 230% of the initial or normal values dependent on the kind of stress. The mean values in 4 cases of Cushing's syndrome were 380% and 1 hour after 25 I.U. ACTH 690% higher than those in normal persons. In normal persons, a well-defined circadian rhythm has been observed.
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Nelson, G. S. (1970). Onchocerciasis. Adv Parasitol, 8, 173–224.
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Collery, L. (1974). Observations of equine animals under farm and feral conditions. Equine Vet J, 6(4), 170–173.
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Krzak, W. E., Gonyou, H. W., & Lawrence, L. M. (1991). Wood chewing by stabled horses: diurnal pattern and effects of exercise. J. Anim Sci., 69(3), 1053–1058.
Abstract: Nine yearling horses, stabled in individual stalls, were used in a trial to determine the diurnal pattern of wood chewing and the effects of exercise on this behavior. The trial was a Latin square design conducted over three 2-wk periods during which each horse was exposed to each of the three following treatments: 1) no exercise (NE), 2) exercise after the morning feeding (AM), and 3) exercise in the afternoon (PM). Horses were fed a complete pelleted feed in the morning and both pelleted feed and long-stemmed hay in the afternoon. Exercise consisted of 45 min on a mechanical walker followed by 45 min in a paddock with bare soil. Each stall was equipped with two untreated spruce boards during each period for wood chewing. Wood chewing was evaluated by videotaping each horse for 22 h during each period, determining the weight and volume of the boards before and after each period, and by visual appraisal of the boards. Intake of trace mineralized salt was also measured. Wood chewing occurred primarily between 2200 and 1200. All measures of wood chewing were correlated when totals for the entire 6 wk were analyzed. When analysis was performed on 2-wk values, videotape results were not correlated with volume or weight loss of boards. Horses chewed more when on the NE treatment (511 s/d) than when on AM or PM (57 and 136 s/d, respectively; P less than .05). Salt intake tended to be greater for NE than for the other treatments (P less than .10).(ABSTRACT TRUNCATED AT 250 WORDS)
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Keay, J. M., Singh, J., Gaunt, M. C., & Kaur, T. (2006). Fecal glucocorticoids and their metabolites as indicators of stress in various mammalian species: a literature review. J Zoo Wildl Med, 37(3), 234–244.
Abstract: Conservation medicine is a discipline in which researchers and conservationists study and respond to the dynamic interplay between animals, humans, and the environment. From a wildlife perspective, animal species are encountering stressors from numerous sources. With the rapidly increasing human population, a corresponding increased demand for food, fuel, and shelter; habitat destruction; and increased competition for natural resources, the health and well-being of wild animal populations is increasingly at risk of disease and endangerment. Scientific data are needed to measure the impact that human encroachment is having on wildlife. Nonbiased biometric data provide a means to measure the amount of stress being imposed on animals from humans, the environment, and other animals. The stress response in animals functions via glucocorticoid metabolism and is regulated by the hypothalamic-pituitary-adrenal axis. Fecal glucocorticoids, in particular, may be an extremely useful biometric test, since sample collection is noninvasive to subjects and, therefore, does not introduce other variables that may alter assay results. For this reason, many researchers and conservationists have begun to use fecal glucocorticoids as a means to measure stress in various animal species. This review article summarizes the literature on many studies in which fecal glucocorticoids and their metabolites have been used to assess stress levels in various mammalian species. Variations between studies are the main focus of this review. Collection methods, storage conditions, shipping procedures, and laboratory techniques utilized by different researchers are discussed.
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Shaw, E. B., Houpt, K. A., & Holmes, D. F. (1988). Body temperature and behaviour of mares during the last two weeks of pregnancy. Equine Vet J, 20(3), 199–202.
Abstract: Average daily core body temperature and behavioural patterns of pregnant mares were studied, in search of definitive signs of parturition within 24 h of the event. Nineteen pony mares were sampled twice daily for core body temperature. A significant temperature drop, averaging 0.1 degrees C (0.2 degrees F) was observed during the day prior to parturition. Between 18.00 h and 06.00 h, during the two weeks before parturition, Thoroughbred and Standardbred mares (n = 52) spent an average 66.8 per cent of their time standing, 27.0 per cent eating, 4.9 per cent lying in sternal recumbency, 1.0 per cent lying in lateral recumbency, and 0.3 per cent walking. On the night before parturition, mares spent significantly less time lying in sternal recumbency than on previous nights and on the night of parturition all behaviour patterns except eating were significantly different from the nights of the two weeks before parturition. There was an increase in walking (5.3 per cent), lying in sternal recumbency (8 per cent) and lying in lateral recumbency (5.3 per cent) whereas standing (53.3 per cent) was decreased. In 58 observed pregnancies, 54 mares (97 per cent) foaled in a recumbent position and 50 mares (86 per cent) foaled between 18.00 h and 06.00 h.
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Bottoms, G. D., Roesel, O. F., Rausch, F. D., & Akins, E. L. (1972). Circadian variation in plasma cortisol and corticosterone in pigs and mares. Am J Vet Res, 33(4), 785–790.
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Kirkpatrick, J. F., Vail, R., Devous, S., Schwend, S., Baker, C. B., & Wiesner, L. (1976). Diurnal variation of plasma testosterone in wild stallions. Biol Reprod, 15(1), 98–101.
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