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      Validating faecal glucocorticoid metabolite analysis in the Virunga mountain gorilla using a natural biological stressor

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          Abstract

          The continued degradation of primate habitat worldwide is forcing many primate populations into small protected forest islands surrounded by high-density human populations. One well-studied example is the critically endangered mountain gorilla ( Gorilla beringei beringei). Decades of monitoring and research on Rwanda's mountain gorillas offer a unique opportunity to use non-invasive endocrine analysis to address pressing questions about the conservation of this endangered population. The aims of our study were as follows: (i) to validate field and laboratory methods for assessing stress through faecal glucocorticoid metabolite (FGM) analysis using inter-social unit interactions as a natural stressor; (ii) to determine the excretion lag times between interactions and detectable stress response in faeces; and (iii) to determine whether there are circadian patterns of FGM excretion. We collected ~6000 faecal samples from 127 known gorillas in 10 habituated groups, monitored by the Dian Fossey Gorilla Fund's Karisoke Research Center over 21 months in 2011 and 2012. Extracted FGMs were measured using a cortisol enzyme immunoassay (R4866; C. J. Munro). Results revealed cause–effect relationships between inter-unit interactions and increased FGMs (relative to individual pre-event samples) between 20 and 140 h after interactions, with the peak most often occurring on day 3. There was no evidence of circadian patterns in FGM concentrations, as previously shown in many species with long gut passage times. However, baseline FGM concentrations were lower in adult males than in adult females, and variation was associated with the collection month, indicating possible seasonal variation. This study provides a biologically validated, field-friendly faecal hormone metabolite extraction and laboratory enzyme immunoassay analysis method for non-invasive monitoring of adrenocortical activity in Virunga mountain gorillas. The methods are useful for future evaluation of a variety of environmental and human-induced potential stressors in this critically endangered population.

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          Physiological stress in ecology: lessons from biomedical research.

          Increasingly, levels of the 'stress hormones' cortisol and corticosterone are being used by ecologists as indicators of physiological stress in wild vertebrates. The amplitude of hormonal response is assumed to correlate with the overall health of an animal and, by extension, the health of the population. However, much of what is known about the physiology of stress has been elucidated by the biomedical research community. I summarize five physiological mechanisms that regulate hormone release during stress that should be useful to ecologists and conservationists. Incorporating these physiological mechanisms into the design and interpretation of ecological studies will make these increasingly popular studies of stress in ecological settings more rigorous.
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            Endocrinology of the stress response.

            The stress response is subserved by the stress system, which is located both in the central nervous system and the periphery. The principal effectors of the stress system include corticotropin-releasing hormone (CRH); arginine vasopressin; the proopiomelanocortin-derived peptides alpha-melanocyte-stimulating hormone and beta-endorphin, the glucocorticoids; and the catecholamines norepinephrine and epinephrine. Appropriate responsiveness of the stress system to stressors is a crucial prerequisite for a sense of well-being, adequate performance of tasks, and positive social interactions. By contrast, inappropriate responsiveness of the stress system may impair growth and development and may account for a number of endocrine, metabolic, autoimmune, and psychiatric disorders. The development and severity of these conditions primarily depend on the genetic vulnerability of the individual, the exposure to adverse environmental factors, and the timing of the stressful events, given that prenatal life, infancy, childhood, and adolescence are critical periods characterized by increased vulnerability to stressors.
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              The concept of allostasis in biology and biomedicine.

              Living organisms have regular patterns and routines that involve obtaining food and carrying out life history stages such as breeding, migrating, molting, and hibernating. The acquisition, utilization, and storage of energy reserves (and other resources) are critical to lifetime reproductive success. There are also responses to predictable changes, e.g., seasonal, and unpredictable challenges, i.e., storms and natural disasters. Social organization in many populations provides advantages through cooperation in providing basic necessities and beneficial social support. But there are disadvantages owing to conflict in social hierarchies and competition for resources. Here we discuss the concept of allostasis, maintaining stability through change, as a fundamental process through which organisms actively adjust to both predictable and unpredictable events. Allostatic load refers to the cumulative cost to the body of allostasis, with allostatic overload being a state in which serious pathophysiology can occur. Using the balance between energy input and expenditure as the basis for applying the concept of allostasis, we propose two types of allostatic overload. Type 1 allostatic overload occurs when energy demand exceeds supply, resulting in activation of the emergency life history stage. This serves to direct the animal away from normal life history stages into a survival mode that decreases allostatic load and regains positive energy balance. The normal life cycle can be resumed when the perturbation passes. Type 2 allostatic overload begins when there is sufficient or even excess energy consumption accompanied by social conflict and other types of social dysfunction. The latter is the case in human society and certain situations affecting animals in captivity. In all cases, secretion of glucocorticosteroids and activity of other mediators of allostasis such as the autonomic nervous system, CNS neurotransmitters, and inflammatory cytokines wax and wane with allostatic load. If allostatic load is chronically high, then pathologies develop. Type 2 allostatic overload does not trigger an escape response, and can only be counteracted through learning and changes in the social structure.
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                Author and article information

                Journal
                Conserv Physiol
                Conserv Physiol
                conphys
                conphys
                Conservation Physiology
                Oxford University Press
                2051-1434
                2016
                30 August 2016
                : 4
                : 1
                : cow029
                Affiliations
                [1 ]The Dian Fossey Gorilla Fund International, Atlanta, GA 30315, USA
                [2 ]Institute for Mind and Biology, University of Chicago , Chicago, IL 60637, USA
                [3 ]Davee Center for Epidemiology & Endocrinology, Lincoln Park Zoo, Chicago, IL 60614, USA
                Author notes
                * Corresponding author: Karisoke Research Center, Karisoke Research Center/Dian Fossey Gorilla Fund International, BP 105, Ruhengeri, Rwanda. Tel: +250 78 835 0016. Email: winnie.eckardt@ 123456gmail.com

                Editor: Steven Cooke

                Article
                cow029
                10.1093/conphys/cow029
                5006093
                27602226
                160ad762-33c1-496e-9931-753184558ee6
                © The Author 2016. Published by Oxford University Press and the Society for Experimental Biology.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 15 December 2015
                : 13 June 2016
                : 22 June 2016
                Funding
                Funded by: Lincoln Park Zoo's Davee Center for Epidemiology & Endocrinology; United States Fish and Wildlife Service;
                Award ID: F12AP01120
                Funded by: Wenner-Gren Foundation, http://dx.doi.org/10.13039/100001388;
                Award ID: 20110146
                Funded by: National Science Foundation Doctoral Dissertation Improvement Grant;
                Award ID: 1122321
                Funded by: Leakey Foundation, http://dx.doi.org/10.13039/100005966;
                Funded by: Judith Harris;
                Categories
                Research Article

                ape,circadian pattern,faecal sample,interaction,lag time
                ape, circadian pattern, faecal sample, interaction, lag time

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