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Palme, R. (2012). Monitoring stress hormone metabolites as a useful, non-invasive tool for welfare assessment in farm animals. Animal Welfare, 21(3), 331–337.
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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Kedzierski, W., Wilk, I., & Janczarek, I. (2014). Physiological response to the first saddling and first mounting of horses: comparison of two sympathetic training methods. Animal Science Papers and Reports, 32(3), 219–228.
Abstract: There is not much research done on the influence of sympathetic training on the emotional reaction
of horses. The aim of the present study was to evaluate the emotional response and the stress level
in horses to two sympathetic training methods: (1) with the use of the “round pen technique” (RP),
and (2) in which the RP was not applied (SH). Twenty two half-bred Anglo-Arab horses (2.5
years ±3 months of age) were subject to an initial training. Eleven horses were randomly included
to the RP method and the other 11 horses for the SH method. Heart rate (HR) and saliva cortisol
concentration were measured as indicators of horse emotional arousal and stress level, respectively.
The HR values were analysed: at rest, during the habituation period, just after the first saddling
and tightening of the girth, during the first time a human leaned over the horse’s back, and during
the mounting of the horse. Saliva samples were taken before and 15 min after each training session
studied. After saddling, the HR occurred significantly higher when the RP technique was used. The
significant increase in saliva cortisol concentration was observed only after the first mounting of
the horse. Generally, the use of the RP technique did not involve more important physiological
reactions in the trained horses than did the SH method.
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Carlsson, H. - E., Lyberg, K., Royo, F., & Hau, J. (2007). Quantification of stress sensitive markers in single fecal samples do not accurately predict excretion of these in the pig. Research in Veterinary Science, 82(3), 423–428.
Abstract: All feces produced during 24 h were collected from five pigs and cortisol and immunoreactive cortisol metabolites (CICM), and IgA were quantified. Within pigs, the concentrations of CICM and IgA varied extensively between random samples obtained from a single fecal dropping, and deviated in most cases significantly from the true concentration measured in total fecal output (CV 6.7–130%). The CICM and IgA contents varied considerably (CV 8.1–114%) within and between individual fecal droppings from the same pig compared to the total fecal excretion. In conclusion, single random samples could not be used to reliably quantify the total fecal concentration or excretion of CICM or IgA in pigs. Analyses of all feces collected during shorter periods than 24 h did not provide an accurate estimate of the daily excretion of CICM. Thus, the concentration of stress sensitive molecules in random single fecal samples as an indicator of animal welfare should be interpreted with prudence.
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Carlstead, K., & Brown, J. L. (2005). Relationships between patterns of Fecal corticoid excretion and behavior, reproduction, and environmental factors in captive black (Diceros bicornis) and white (Ceratotherium simum) rhinoceros. Zoo Biol., 24(3), 215–232.
Abstract: Mortality is high in zoo-housed black rhinoceros (Diceros bicornis), and the reproductive rates of captive white rhinoceros (Ceratotherium simum) are unsustainably low. To determine the possible role of stress in the causation of these problems, we analyzed weekly fecal samples collected for 1 year from black (10 males and 16 females) and white (six males and 13 females) rhinoceroses at 16 zoos for corticoid metabolite concentrations. Fecal corticoid profiles were examined in relation to behavior as rated by keepers in a questionnaire, luteal phase ovarian cycles of females (Brown et al., 2001), and socioenvironmental factors. We compared individual fecal corticoid profiles by examining hormone means and variability (i.e., standard deviation (SD) and coefficient of variation (CV)). For the black rhinos, higher mean corticoid concentrations were found at zoos where rhinos were maintained in enclosures that were exposed to the public around a greater portion of the perimeter. Higher variability in corticoid excretion was correlated with higher rates of fighting between breeding partners and higher institutional mortality rates. Black rhino pairs that were kept separated exhibited lower corticoid variability and less fighting activity when they were introduced during female estrous periods compared to pairs that were kept together every day. For white rhinos, significantly lower mean corticoids were found for individuals that rated higher on “friendliness to keeper.” Higher corticoid variability was found in noncycling as compared to cycling white rhino females. Noncycling females exhibited higher rates of stereotypic pacing and lower frequencies of olfactory behaviors. Interindividual differences in mean corticoids in both species appeared to be related to responsiveness to humans, whereas corticoid variability was related to intraspecific social relationships. More importantly, high corticoid variability appeared to be an indicator of chronic or “bad” stress, because of its association with potentially deleterious consequences in each species (i.e., fighting and mortality (black rhino), and reproductive acyclicity (white rhino)). Our results provide evidence that social stressors may cause chronic stress in black and white rhinos, and that this contributes to the captive-population sustainability problems observed in each species. Zoo Biol 0:1–18, 2005. © 2005 Wiley-Liss, Inc.
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Meehan, C. L., & Mench, J. A. (2007). The challenge of challenge: Can problem solving opportunities enhance animal welfare? Appl. Anim. Behav. Sci., 102(3-4), 246–261.
Abstract: Cognitive mechanisms are an important part of the organization of the behavior systems of animals. In the wild, animals regularly face problems that they must overcome in order to survive and thrive. Solving such problems often requires animals to process, store, retrieve, and act upon information from the environment--in other words, to use their cognitive skills. For example, animals may have to use navigational, tool-making or cooperative social skills in order to procure their food. However, many enrichment programs for captive animals do not include the integration of these types of cognitive challenges. Thus, foraging enrichments typically are designed to facilitate the physical expression of feeding behaviors such as food-searching and food consumption, but not to facilitate complex problem solving behaviors related to food acquisition. Challenging animals by presenting them with problems is almost certainly a source of frustration and stress. However, we suggest here that this is an important, and even necessary, feature of an enrichment program, as long as animals also possess the skills and resources to effectively solve the problems with which they are presented. We discuss this with reference to theories about the emotional consequences of coping with challenge, the association between lack of challenge and the development of abnormal behavior, and the benefits of stress (arousal) in facilitating learning and memory of relevant skills. Much remains to be done to provide empirical support for these theories. However, they do point the way to a practical approach to improving animal welfare--to design enrichments to facilitate the cognitive mechanisms which underlie the performance of complex behaviors that cannot be performed due to the restrictions inherent to the captive environment.
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Cayado, P., Munoz-Escassi, B., Dominguez, C., Manley, W., Olabarri, B., Sanchez de la Muela, M., et al. (2006). Hormone response to training and competition in athletic horses. Equine Vet J Suppl, (36), 274–278.
Abstract: REASONS FOR PERFORMING STUDY: It is recognised that the amount of psychological stress that an animal encounters determines the degree of response of the hypothalamic-pituitary-adrenal (HPA) axis. In human athletes, the added emotive stress of competition is an important element in the adrenal response. The aim of this study was to examine the effect of show-jumping as well as dressage on stress levels by comparing horses' stress response at a horse show compared to their familiar home. METHODS: Fifty-one horses involved in competition were used. EDTA blood samples were collected before exercise, upon arrived to the schooling area (control), and k over a jump or dressage course. After sampling, plasma was separated and stored at -80 degrees C until determinations of cortisol and ACTH were performed. Fourteen healthy horses not involved in competition were used as control group. RESULTS: Competition induced a significant increase in cortisol and ACTH responses in both, jumping and dressage horses and this effect was more apparent in dressage horses. When horses were most experienced, cortisol and ACTH responses were much lower. CONCLUSION: This study shows that competition elicits a classic physiological stress response in horses and that different training programmes induce different responses. It suggests that horses involved in competition can provide a good model to study the exercise-induced stress response.
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Innes, L., & McBride, S. (2008). Negative versus positive reinforcement: An evaluation of training strategies for rehabilitated horses. Appl. Anim. Behav. Sci., 112(3–4), 357–368.
Abstract: Rescued equids are often exposed to rehabilitation and training (or retraining) programmes to improve their physical and psychological well-being as well as to facilitate the re-homing process. Training uses either positive or negative reinforcement learning procedures and it is considered here that, there may be welfare implications associated with using the latter technique as it has the potential to overlay acute stress on animals with a chronic stress life history. The aim of this study, therefore, was to compare these training strategies (negative versus positive reinforcement) on equine behaviour and physiology as the first step in establishing an optimal rehabilitation approach (from a welfare perspective) for equids that have been subjected to chronic stress in the form of long-term neglect/cruelty. Over a 7-week period, 16 ponies (aged 6–18 months) were trained using either positive (‘positive’) (n = 8) or negative reinforcement (‘negative’) (n = 8) techniques to lead in hand, stand to be groomed, traverse an obstacle course and load into a trailer. Heart rate was measured (5 s intervals) on days 1 and 4 of each training week, ‘Pre’- (1 h), ‘During’ (0.5 h) and ‘Post’- (1 h) training session. Ethograms (10.00–20.00 h) outside of the training period were also compiled twice weekly. In addition, weekly arena tests (as a measure of reactivity) were also performed 1 week before and during the 7 weeks of training. Results showed significant differences between the two training schedules for some measures during the latter stages of the trial and suggested that animals trained under a positive reinforcement schedule were more motivated to participate in the training sessions and exhibited more exploratory or ‘trial and error’ type behaviours in novel situations/environments. In this context, the incorporation of positive reinforcement schedules within a rehabilitation programme may be of benefit to the animal from a welfare perspective.
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Houpt, K. A., Eggleston, A., Kunkle, K., & Houpt, T. R. (2000). Effect of water restriction on equine behaviour and physiology. Equine Vet J, 32(4), 341–344.
Abstract: Six pregnant mares were used to determine what level of water restriction causes physiological and/or behavioural changes indicative of stress. Nonlegume hay was fed ad libitum. During the first week of restriction, 5 l water/100 kg bwt was available, during the second week 4 l/100 kg bwt and, during the third week, 3 l/100 kg bwt. Ad libitum water intake was 6.9 l/100 kg bwt; at 3 l/100 kg bwt water intake was 42% of this. Daily hay intake fell significantly with increasing water restriction from 12.9 +/- 0.75 kg to 8.3 +/- 0.54 kg; bodyweight fell significantly for a total loss of 48.5 +/- 8.3 kg in 3 weeks. Daily blood samples were analysed; osmolality rose significantly with increasing water restriction from 282 +/- 0.7 mosmols/kg to 293.3 +/- 0.8 mosmols/kg bwt, but plasma protein and PCV did not change significantly. Cortisol concentrations fell from 8.1 ng/ml to 6.4 ng/ml over the 3 week period. Aldosterone fell from 211.3 +/- 74.2 pg/ml to 92.5 +/- 27.5 pg/ml at the end of the first week. The behaviour of 4 of the 6 mares was recorded 24 h/day for the duration of the study. The only significant difference was in time spent eating, which decreased with increasing water restriction from 46 +/- 3% to 30 +/- 3%. It is concluded that water restriction to 4 l/100 kg bwt dehydrates pregnant mares and may diminish their welfare, but is not life- or pregnancy-threatening.
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Shanahan, S. (2003). Trailer loading stress in horses: behavioral and physiological effects of nonaversive training (TTEAM). J Appl Anim Welf Sci, 6(4), 263–274.
Abstract: Resistance in the horse to trailer loading is a common source of stress and injury to horses and their handlers. The objective of this study was to determine whether nonaversive training based on the Tellington-Touch Equine Awareness Method (TTEAM; Tellington-Jones &Bruns, 1988) would decrease loading time and reduce stress during loading for horses with a history of reluctance to load. Ten horses described by their owners as “problem loaders” were subjected to pretraining and posttraining assessments of loading. Each assessment involved two 7-min loading attempts during which heart rate and saliva cortisol were measured. The training consisted of six 30-min sessions over a 2-week period during which the horse and owner participated in basic leading exercises with obstacles simulating aspects of trailering. Assessment showed heart rate and saliva cortisol increased significantly during loading as compared to baseline (p <.001 and p <.05, respectively). Reassessment after training showed a decrease in loading time (p <.02), reduced heart rate during loading (p <.002), and reduced saliva cortisol as compared to pretraining assessments. Seven “good loaders” also were subject to loading assessment for physiological comparison. Increases in heart rate during loading were significantly higher in the good loaders (p <.001). Nonaversive training simulating aspects of loading may effectively reduce loading time and stress during loading for horses with a history of resistance to trailer loading.
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Bachmann, I., Bernasconi, P., Herrmann, R., Weishaupt, M. A., & Stauffacher, M. (2003). Behavioural and physiological responses to an acute stressor in crib-biting and control horses. Appl. Anim. Behav. Sci., 82(4), 297–311.
Abstract: The responses of eleven pairs of crib-biting and non-crib-biting horses (controls) to an arousal-inducing stimulus were studied. Video-observation of the horses revealed that crib-biting horses spent between 10.4 and 64.7% of their stabling time performing the stereotypy. During the first 2 days of an experimental period, the horses were conditioned to receive food from a special bucket. On the third day the food bucket was presented, but the horses were not allowed to feed. Arousal behaviour and crib-biting intensity as well as plasma cortisol concentration, heart rate (HR) and heart rate variability (HRV) were recorded at rest, and during and after presentation of the food stimulus. The stimulus induced a significant increase of HR and arousal behaviour in crib-biters and in controls, whereas the crib-biting frequency decreased. Power spectral analysis of the HRV revealed significant differences between crib-biters and controls at rest: crib-biters had a lower vagal tone (high frequency component, HF) and a higher sympathetic tone (low frequency component, LF) than controls. The lower basal parasympathetic activity might be an indication why crib-biting horses, in contrast to the controls, showed neither a significant decrease of the HF component during presentation of the food stimulus nor an increase of the HF component after presentation. Thus, there might be differences in the tuning of the autonomous nervous system and of the stress reactivity in crib-biting and in control horses. The results suggest that the crib-biting horses are more stress sensitive and physiologically and psychologically less flexible than the control horses.
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