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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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Skandakumar, S., Stodulski, G., & Hau, J. (1995). Salivary IgA: a Possible Stress Marker In Dogs. In Animal Welfare (Vol. 4, pp. 339–350).
Abstract: Stress in humans has been reported to be associated with a decrease in the salivary immunoglobulin A (s-IgA) levels enabling the possible use of s-IgA to assess stress. Prolonged stress, if reliably assessed in a non-invasive manner, may be used to assess animal welfare. This study analysed groups of dogs undergoing physical and temperamental training and s-IgA levels were measured by rocket immunoelectrophoresis in prospective samples. Behavioural assessment was carried out and cortisol levels in saliva were measured by ELISA. A significant negative correlation (P < 0.007) between the logarithmic cortisol concentrations and s-IgA levels in saliva was recorded. The behavioural assessment of the dogs agreed well with the biochemical markers. It is concluded that IgA levels in saliva may be a useful marker of dog well-being and that stress results in decreased s-IgA levels.
Keywords: Animal Welfare; Behaviour; Cortisol; Dog; Salivary Iga (S-Iga); Stress; Well-Being
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Dalmau, A., Ferret, A., Chacon, G., & Manteca, X. (2007). Seasonal Changes in Fecal Cortisol Metabolites in Pyrenean Chamois. J Wildl Manag, 71(1), 190–194.
Abstract: We studied seasonal changes in fecal cortisol metabolites (FCM), which have been widely used as indicators of stress, in a population of Pyrenean chamois (Rupicapra pyrenaica pyrenaica) in the Cadí Range of northeastern Spain. We collected fecal samples from 2001 to 2003 in 3 particular locations with different altitudes and male or female presence, and we analyzed them for FCM and fecal nitrogen as an indicator of diet quality. We observed a clear seasonal pattern, with the highest FCM in winter, and we obtained correlations between FCM and monthly mean minimum temperatures and fecal nitrogen. We observed no effects of tourism presence, trophy hunting, or rut season on FCM. Analysis of cortisol metabolites in feces can be a good measure of winter stress in Pyrenean chamois.
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Flauger, B., Krueger, K., Gerhards, H., & Möstl, E. (2010). Simplified method to measure glucocorticoid metabolites in faeces of horses. Vet Res Comm, 34(2), 185–195.
Abstract: Glucocorticoids or their metabolites can be measured in several body fluids or excreta, including plasma, saliva, urine and faeces. In recent years the measurement of glucocorticoid metabolites (GCMs) in faeces has gained increasing attention, because of its suitability for wild populations. In horses, however, the group-specific enzyme immunoassay described so far has a limited racticability due to its complex extraction procedure. Therefore, we tested the applicability of
other enzyme immunoassays for glucocorticoid metabolites. The present study clearly proved that an enzyme immunoassay (EIA) for 11-oxoetiocholanolone using 11-oxoetiocholanolone-17-CMO: BSA (3α,11-oxo-A EIA) as antigen showed high amounts of immunoreactive substances. Therefore it was possible to use just a small amount of the supernatant of a methanolic suspension of faeces. The results correlated well with the already described method for measuring GCMs in horse faeces, i.e. analysing the samples with an EIA after a two step clean up procedure of the samples (Merl et al. 2000). In addition, the 3α,11-oxo-A EIA has the advantage of providing a bigger difference between baseline values and peak values after ACTH stimulation. The new assay increased the accuracy of the test, lowered the expenses per sample, and storing samples at room temperature after collection was less critical than with other assays investigated in our study. This is a big advantage both in the field of wildlife management of equids and in the field of equestrian sports and it shows the importance of choosing an assay which is in good accordance with the metabolites excreted in a given species. Keywords: ACTH challenge; enzyme immunoassay; stress behaviour; cortisol
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Hoshaw, B. A., Evans, J. C., Mueller, B., Valentino, R. J., & Lucki, I. (2006). Social competition in rats: Cell proliferation and behavior. Behav. Brain. Res., 175(2), 343–351.
Abstract: Behavioral and physiological changes were studied following prolonged exposure to social competition in pairs of non-food-deprived rats competing daily for a limited supply of graham cracker crumbs. Stable dominant-subordinate relationships developed in most pairs, as measured by feeding time, which were maintained over a 5-6-week study period. In other behavioral tests, subordinates demonstrated a decreased latency to immobility in the forced swim test compared with dominants, but no difference in locomotor activity. Subordinates had increased bladder size, decreased adrenal gland size, and a 35% reduction of hippocampus cell proliferation compared with the dominant member. Therefore, prolonged social competition, based on restricted access to palatable substances, produced hierarchies among individuals that were associated with differences in behavior, physiology and hippocampal cell proliferation.
Keywords: Social stress; Depression; Forced swim test; Neurogenesis
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Creel, S. (2001). Social dominance and stress hormones. Trends. Ecol. Evol, 16(9), 491–497.
Abstract: In most cooperatively breeding birds and mammals, reproductive rates are lower for social subordinates than for dominants, and it is common for reproduction in subordinates to be completely suppressed. Early research conducted in captivity showed that losing fights can increase glucocorticoid (GC) secretion, a general response to stress. Because GCs can suppress reproduction, it has been widely argued that chronic stress might underlie reproductive suppression of social subordinates in cooperative breeders. Contradicting this hypothesis, recent studies of cooperative breeders in the wild show that dominant individuals have elevated GCs more often than do subordinates. The findings that elevated GCs can be a consequence of subordination or a cost of dominance complicate the conventional view of social stress, with broad ramifications for the evolution of dominance and reproductive suppression.
Keywords: Dominance; rank; stress; glucocorticoids; cooperative breeding; sociality; behavioural endocrinology; mammals
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Griffin, A. S. (2008). Social learning in Indian mynahs, Acridotheres tristis: the role of distress calls. Anim. Behav., 75(1), 79–89.
Abstract: Socially acquired predator avoidance is a phenomenon in which individuals acquire an avoidance response towards an initially neutral stimulus after they have experienced it together with the antipredator signals of social companions. Earlier research has established that alarm calls used for intraspecific communication are effective stimuli for triggering acquisition. However, animals produce a large range of other antipredator responses that might engage antipredator learning. Here, I examine the effects of conspecific distress calls, a signal that is produced by birds when restrained by a predator, and that appears to be directed towards predators, rather than conspecifics, on predator avoidance learning in Indian mynahs, Acridotheres tristis. Distress calls reflect high levels of alarm in the caller and should, therefore, mediate robust learning. Experiment 1 revealed that subjects performed higher rates of head movements in response to a previously unfamiliar avian mount after it had been presented simultaneously with playbacks of conspecific distress vocalizations. Experiment 2 revealed that increased rates of head saccades resembled the spontaneous response evoked by a novel stimulus more closely than it resembled the response evoked by a perched raptor, suggesting that distress calls inculcated a visual exploratory response, rather than an antipredator response. While it is usually thought that the level of acquisition in learners follows a simple relationship with the level of alarm shown by demonstrators, the present results suggest that this relationship may be more complex. Antipredator signals with different functions may have differential effects on learners.
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VanDierendonck, M. C. (2006). Social relationships in a group of horses without a mature stallion (Vol. Chapter 4). Universiteit Utrecht.
Abstract: Feral horses are social animals, which have to rely on survival strategies centered on the formation of cohesive social bonds within their bands. Many problems in the husbandry of social animals such as horses, are due to the fact that the limits of their adaptive abilities are exceeded. Evidence suggests that the fundamental social characteristics of domestic horses have remained relatively unchanged. The social structure, social strategies and social interactions were investigated (3 non-consecutive years, 24 hr per day for several weeks) in long term established groups of domestic horses (mares and geldings of all ages) and a few small introduced groups, kept in (semi)natural environments. The general aim was to investigate the social needs of domestic horses. The social life of domestic horses was characterised by long lasting bonds with preferred partners which were established and maintained by allogrooming, play, proximity and dominance behaviours. Bonding partners were mainly found within the same sex-age group, but adult geldings also bonded with sub-adult mares and geldings. Adult mares were clustered in a group, while the other animals formed a second group. Among the adult mares, subgroups according to reproductive state were formed. Individuals regulated their social network by interfering with interactions between other members of the herd, which in itself is complex. An intervention is a behavioural action of one animal that actively interferes with an ongoing interaction between a dyad with the apparent aim of altering that interaction. This was verified by post-hoc analyses of disturbed and undisturbed interactions. Interventions in allogrooming or play were performed significantly more often when at least one member of the initial dyad was a preferred partner of, or familiar to (within the small introduced bands) the intervener. The stronger the preferred association in allogrooming between the intervener and member(s) of the initial dyad, the higher the probability the intervener would displace one initial member and continue allogrooming with the other. Just five behaviours were extracted which reliably reflected the dominance relations among horses. Aggression with the hind quarters was used both offensively and defensively and therefore not suitable as a reliable parameter. Individual dominance relationships were related to social experience. The implications of these findings for horse husbandry were assessed. It is argued that the execution of affiliative behaviours may be rewarding in itself, and therefore always will be a highly motivated behaviour. It is shown that social positive physical interactions (allogrooming, play) with other horses is an ethological need and therefore indispensable in modern husbandry systems. Ethological needs are so important for the animal that husbandry systems that lack the possibilities to execute such behaviours will cause chronic stress. It is concluded that all horses need physical social contact, and that horses, which lack appropriate social learning experiences during ontogeny, may be hampered in their social functioning later in life. Solutions for problems, including dominance problems, in individual social housing and group housing are presented.
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Palme, R., Touma, C., Arias, N., Dominchin, M.N., & Lepschy, M. (2013). Steroid extraction: Get the best out of faecal samples. Wien Tierärztl Monat – Vet Med Austria, 100, 238–246.
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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h:, M., Lévy, F., Fortin, M., Leterrier, C., & LansadLansade, L. (2013). Stress and temperament affect working memory performance for disappearing food in horses, Equus caballus. Animal Behaviour, 86(6), 1233–1240.
Abstract: In the present study, we sought to determine the influence of stress and temperament on working memory for disappearing food in horses. After assessment of five dimensions of temperament, we tested working memory of horses using a delayed-response task requiring a choice between two food locations. Delays ranging from 0 to 20 s were tested. The duration of working memory for disappearing food was first characterized without stressors (N = 26). The horses were then divided into two groups and their performance was assessed under stressful (exposure to acute stressors prior to testing, N = 12) or control conditions (N = 12). Results showed that the duration of working memory for disappearing food lasted at least 20 s under nonstressful conditions, and that under stressful conditions this duration lasted less than 12 s. This stress-induced impairment confirms in a nonrodent species that working memory performance is very sensitive to exposure to stressors. In addition, working memory performance in horses is influenced by the temperamental dimension of fearfulness according to the state of stress: fearful horses showed better performance under control conditions and worse performance under stressful conditions than nonfearful horses. These findings are discussed in the context of the Yerkes–Dodson law of stress and performance.
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