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Use of feather corticosterone analysis in avian research has become a widespread and powerful tool, however, there has been limited work on assessing the robustness of the methodology. Through several assay validations we address current issues and provide suggestions for further refinement of the technique, Feather corticosterone measurement is becoming a widespread tool for assessing avian physiology. Corticosterone is deposited into feathers during growth and provides integrative and retrospective measures of an individual’s hypothalamic–pituitary–adrenal (HPA) axis function. Although researchers across disciplines have been measuring feather corticosterone for the past decade, there are still many issues with the extraction and measurement of corticosterone from feathers. In this paper, we provide several directives for refining the methodology for feather hormone analysis. We compare parallelism between the standard curve and serially diluted feather tissue from wild turkeys, Canada jays, and black-capped chickadees to demonstrate the wide applicability across species. Through a series of validations, we compare methods for feather preparation, sample filtration and extract reconstitution prior to corticosterone quantification using a radioimmunoassay. Higher corticosterone yields were achieved following pulverization of the feather however, more variation between replicates was observed. Removal of the rachis also increased the amount of corticosterone detected per unit mass while glass versus paper filters had no effect, and using ethanol in the reconstution buffer decreased intra-assay variation. With these findings and continued methodological refinement, feather corticosterone has the potential to be a powerful tool for both ecologists and physiologists working with historical and contemporary specimens.

A bibliometric analysis of contemporary peer-reviewed literature was used to examine the overlap between conservation and physiology. Although the term “conservation physiology” rarely appears in papers that combine the disciplines, they are indeed being integrated in recent years and we expect increased integration in the near future., Contemporary conservation science benefits from the perspectives of a variety of different disciplines, including a recent synergy with physiology, an interface known as ‘conservation physiology’. To evaluate the degree of interaction between conservation and animal/plant physiology, we conducted three bibliometric analyses. We first pursued the use of the term ‘conservation physiology’ since its first definition in 2006 to determine how frequently it has been used and in which publications. Secondly, we evaluated the occurrence of conservation terms in animal and plant physiology journals, physiological terms in conservation journals, and a combination of terms in ecology journals. Thirdly, we explored trends in a subset of conservation physiology articles published between 2006 and 2012. We identified a surge in the use of the term ‘conservation physiology’ in 2012, after only a slow increase in usage between 2006 and 2011. Conservation journals tend to have been significantly more active in publishing conservation physiology than animal physiology, plant physiology or ecology journals. However, we found evidence that ecology and animal physiology journals began to incorporate more conservation physiology after 2006, while conservation- and plant physiology-themed journals did not. Among 299 conservation physiology articles that we identified, vertebrate taxa have been over-represented in conservation physiology compared with their relative taxonomic abundance, invertebrate taxa have been under-represented, and plants have been represented in proportion to their relative taxonomic abundance; however, those findings are reasonably consistent with publication trends in conservation biology. Diffuse distribution of conservation physiology papers throughout the literature may have been a barrier to the growth of the subdiscipline when the interface was emerging. The introduction of the focused journal Conservation Physiology in 2013 may address that deficiency. Moreover, development of a unifying framework could help to aggregate knowledge and attract potential contributors by highlighting and facilitating access to and application of conservation physiology.

This paper explores how biological responses can inform conservation strategies of threatened giant clams through global change, including climate change, global warming, marine heatwaves and ocean acidification. Understanding responses to environmental stressors can help project future distributions of suitable giant clam habitat for protection and use in selective breeding programmes., Giant clams are threatened by overexploitation for human consumption, their valuable shells and the aquarium trade. Consequently, these iconic coral reef megafauna are extinct in some former areas of their range and are included in the International Union for Conservation of Nature (IUCN) Red List of Threatened Species and Convention on International Trade in Endangered Species of Wild Fauna and Flora. Now, giant clams are also threatened by rapid environmental change from both a suite of local and regional scale stressors and global change, including climate change, global warming, marine heatwaves and ocean acidification. The interplay between local- to regional-scale and global-scale drivers is likely to cause an array of lethal and sub-lethal effects on giant clams, potentially limiting their depth distribution on coral reefs and decreasing suitable habitat area within natural ranges of species. Global change stressors, pervasive both in unprotected and protected areas, threaten to diminish conservation efforts to date. International efforts urgently need to reduce carbon dioxide emissions to avoid lethal and sub-lethal effects of global change on giant clams. Meanwhile, knowledge of giant clam physiological and ecological responses to local–regional and global stressors could play a critical role in conservation strategies of these threatened species through rapid environmental change. Further work on how biological responses translate into habitat requirements as global change progresses, selective breeding for resilience, the capacity for rapid adaptive responses of the giant clam holobiont and valuing tourism potential, including recognizing giant clams as a flagship species for coral reefs, may help improve the prospects of these charismatic megafauna over the coming decades.

Some eight years ago Martin Wikelski and I started musing about how physiological knowledge and tools could be used to understand conservation problems. We were certainly not the first to do so, but we were able to codify the idea by coin ing the term ‘conservation physiology’ (Wikelski and Cooke, 2006), which has been embraced by many. When the leadership at the Society of Experimental Biology (SEB), including Tony Farrell, Craig Franklin, and Paul Hutchinson, embarked on a plan to launch a new journal, the topic of conservation physiology quickly rose to the top, given its timeliness and potential to engage both plant and animal researchers. That was nearly two years ago. Today I am thrilled to launch the second volume of the journal Conservation Physiology, published jointly by Oxford University Press (OUP) and the SEB. Here I briefly reflect on the first year (2013) of contributions to Conservation Physiology and consider what can be expected in 2014 and beyond. The Conservation Physiology submission portal opened quietly in December of 2012 and, with negligible advertising, we started receiving submissions. The inaugural paper (Cooke et al., 2013) was published in March, with the official launch of the journal occurring with great fanfare at the SEB Annual Meeting in Valencia, Spain in July. As of December 2013, Volume 1 had over 30 papers published, with another handful of accepted articles in production. The content is remarkably diverse, spanning taxa including plants (Funk

Freshwater fish must maintain the concentration of salts in their blood within a narrow range, but this status is challenged by the prevailing water chemistry and its fluctuations. This paper provides a framework for understanding how fish in nature will respond to forecasted future changes in water chemistry., Alterations in water chemistry can challenge resident fish species. More specifically, chemical changes that disrupt ion balance will negatively affect fish health and impact physiological and ecological performance. However, our understanding of which species and populations are at risk from ionoregulatory disturbances in response to changing freshwater environments is currently unclear. Therefore, we propose a novel framework for incorporating ionoregulatory physiology into conservation management of inland fishes. This framework introduces the concepts of fundamental chemical niche, which is the tolerable range of chemical conditions for a given species based on laboratory experiments, and realized chemical niche, which is the range of chemical conditions in which a species resides based on distribution surveys. By comparing these two niches, populations that may be at risk from ionoregulatory disturbances and thus require additional conservation considerations can be identified. We highlight the potential for commonly measured ionoregulatory traits to predict fundamental and realized chemical niches but caution that some traits may not serve as accurate predictors despite being important for understanding ionoregulatory mechanisms. As a sample application of our framework, the minimum pH distribution (realized niche) and survival limit pH (fundamental niche) of several North American fishes were determined by systematic review and were compared. We demonstrate that ionoregulatory capacity is significantly correlated with a realized niche for many species, highlighting the influence of ionoregulatory physiology on fish distribution patterns along chemical gradients. Our aim is that this framework will stimulate further research in this field and result in a broader integration of physiological data into conservation management decisions for inland waters.

This paper reviews applications of dynamic energy budget (DEB) theory to conservation issues. We discuss how the DEB framework, through its mechanistic nature, its universality and the wide range of outputs it provides, is well equipped to tackle current and future challenges in the conservation of species., The contribution of knowledge, concepts and perspectives from physiological ecology to conservation decision-making has become critical for understanding and acting upon threats to the persistence of sensitive species. Here we review applications of dynamic energy budget (DEB) theory to conservation issues and discuss how this theory for metabolic organization of all life on earth (from bacteria to whales) is well equipped to support current and future investigations in conservation research. DEB theory was first invented in 1979 in an applied institution for environmental quality assessment and mitigation. The theory has since undergone extensive development and applications. An increasing number of studies using DEB modelling have provided valuable insights and predictions in areas that pertain to conservation such as species distribution, evolutionary biology, toxicological impacts and ecosystem management. We discuss why DEB theory, through its mechanistic nature, its universality and the wide range of outcomes it can provide represents a valuable tool to tackle some of the current and future challenges linked to maintaining biodiversity, ensuring species survival, ecotoxicology, setting water and soil quality standards and restoring ecosystem structure and functioning in a changing environment under the pressure of anthropogenic driven changes.

Populus fremontii (Fremont cottonwood) is recognized as one of the most important foundation tree species in the southwestern USA and northern Mexico because of its ability to structure communities across multiple trophic levels, drive ecosystem processes and influence biodiversity via genetic-based functional trait variation. However, the areal extent of P. fremontii cover has declined dramatically over the last century due to the effects of surface water diversions, non-native species invasions and more recently climate change. Consequently, P. fremontii gallery forests are considered amongst the most threatened forest types in North America. In this paper, we unify four conceptual areas of genes to ecosystems research related to P. fremontii’s capacity to survive or even thrive under current and future environmental conditions: (i) hydraulic function related to canopy thermal regulation during heat waves; (ii) mycorrhizal mutualists in relation to resiliency to climate change and invasion by the non-native tree/shrub, Tamarix; (iii) phenotypic plasticity as a mechanism for coping with rapid changes in climate; and (iv) hybridization between P. fremontii and other closely related Populus species where enhanced vigour of hybrids may preserve the foundational capacity of Populus in the face of environmental change. We also discuss opportunities to scale these conceptual areas from genes to the ecosystem level via remote sensing. We anticipate that the exploration of these conceptual areas of research will facilitate solutions to climate change with a foundation species that is recognized as being critically important for biodiversity conservation and could serve as a model for adaptive management of arid regions in the southwestern USA and around the world.

Lay summary: A review that compares changes in body mass, glucocorticoid and sympathetic responses, and reproductive and immune function, in wild animals recently introduced into captivity to their wild counterparts. Conclusion is that captivity can be a powerful chronic stressor that may be possible to mitigate, but the impact is highly species-specific., Wild animals are brought into captivity for many reasons—conservation, research, agriculture and the exotic pet trade. While the physical needs of animals are met in captivity, the conditions of confinement and exposure to humans can result in physiological stress. The stress response consists of the suite of hormonal and physiological reactions to help an animal survive potentially harmful stimuli. The adrenomedullary response results in increased heart rate and muscle tone (among other effects); elevated glucocorticoid (GC) hormones help to direct resources towards immediate survival. While these responses are adaptive, overexposure to stress can cause physiological problems, such as weight loss, changes to the immune system and decreased reproductive capacity. Many people who work with wild animals in captivity assume that they will eventually adjust to their new circumstances. However, captivity may have long-term or permanent impacts on physiology if the stress response is chronically activated. We reviewed the literature on the effects of introduction to captivity in wild-caught individuals on the physiological systems impacted by stress, particularly weight changes, GC regulation, adrenomedullary regulation and the immune and reproductive systems. This paper did not review studies on captive-born animals. Adjustment to captivity has been reported for some physiological systems in some species. However, for many species, permanent alterations to physiology may occur with captivity. For example, captive animals may have elevated GCs and/or reduced reproductive capacity compared to free-living animals even after months in captivity. Full adjustment to captivity may occur only in some species, and may be dependent on time of year or other variables. We discuss some of the methods that can be used to reduce chronic captivity stress.

Identifying biomarkers of well-being will be beneficial to our understanding of animal welfare. We developed and validated an assay for measuring Asian elephant immunoglobulin A (IgA) in feces, saliva, urine and serum as a potential welfare measure, and show that longitudinal assessments are necessary because of high intra-individual variability., Additional measures of well-being would be beneficial to the management of a variety of species in human care, including elephants. Immunoglobulin A (IgA) is an immune protein associated with pathogen defense, which has been demonstrated to decrease during times of stress, and increase in response to positive stimuli. This paper describes the development and validation of an enzyme immunoassay (EIA) for the quantification of Asian elephant (Elephas maximus) IgA in feces, saliva, urine, and serum. Samples were collected weekly from four females for 6 months to assess IgA and glucocorticoid (GC) concentrations, establish relationships between these two biomarkers, and determine variability in IgA within and between individuals, and across sample types. IgA was quantified in all four sample types, although urinary concentrations were low and sometimes undetectable in individual samples. Concentrations were highly variable within and between individuals, with fecal, salivary and serum IgA, and fecal, salivary and urinary GCs all differing significantly across individuals. Contrary to previous findings, IgA and GC were generally not correlated. Serum IgA was less variable within individuals, with the exception of one female that experienced a brief illness during the study. However, marked inter-individual differences were still apparent. When data from all individuals were combined, fecal IgA was significantly predicted by salivary and urinary IgA; however, this relationship did not hold when individuals were analyzed separately. Analysis of a fifth female that exhibited a more severe systemic illness demonstrated clear increases in fecal IgA and GC, suggesting these may also be useful health biomarkers. Further investigation is needed to determine what sample type is most reflective of biological state in elephants, and how IgA concentrations are associated with health and positive and negative welfare states. Based on observed variability, a longitudinal approach likely will be necessary to use IgA as a measure of well-being.

There are limited studies reporting the use of cryopreserved sperm in amphibians, and fewer reporting the generation of sexually mature individuals. This paper gives proof of concept to the value of genome banking for amphibian conservation by demonstrating in one amphibian species, the Eastern dwarf tree frog (Litoria fallax), that offspring produced from cryopreserved sperm are capable of achieving metamorphosis and reaching sexually maturity., Amphibians are the most threatened vertebrate class globally based on recent rates of decline and extinction. Sperm cryopreservation and other assisted reproductive technologies have the potential to help manage small and threatened populations and prevent extinctions. There are a growing number of reports of recovery of amphibian sperm after cryopreservation, but relatively few published reports of amphibian embryos generated from frozen sperm developing beyond metamorphosis to the adult stage and achieving sexual maturation. In this study on the Eastern dwarf tree frog (Litoria fallax), a temperate amphibian species from eastern Australia, a small number of viable metamorphs and one sexually mature male frog (itself producing sperm) were produced from cryopreserved sperm, demonstrating the capacity of embryos generated from cryopreserved sperm to complete the life cycle to sexual maturity. Low progression rates between developmental stages were not deemed to be due to effects of cryopreservation, since control embryos from unfrozen sperm had a similarly low progression rate through development.

Atmospheric nitrogen deposition resulting from anthropic activities is a major threat to global biodiversity. Given high costs of deploying automated monitoring networks, this paper makes the case for the use of plant biomonitors, such as mosses, grasses, trees and epiphytes to characterize atmospheric nitrogen pollution., Atmospheric nitrogen deposition is the third largest cause of global biodiversity loss, with rates that have more than doubled over the past century. This is especially threatening for tropical regions where the deposition may soon exceed 25 kg of N ha−1 year−1, well above the threshold for physiological damage of 12–20 kg of N ha−1 year−1, depending on plant species and nitrogenous compound. It is thus urgent to monitor these regions where the most diverse biotas occur. However, most studies have been conducted in Europe, the USA and recently in China. This review presents the case for the potential use of biological organisms to monitor nitrogen deposition, with emphasis on tropical plants. We first present an overview of atmospheric chemistry and the nitrogen metabolism of potential biomonitors, followed by a framework for monitoring nitrogen deposition based on the simultaneous use of various functional groups. In particular, the tissue nitrogen content responds to the rate of deposition, especially for mosses, whose nitrogen content increases by 1‰ per kilogram of N ha−1 year−1. The isotopic signature, δ15N, is a useful indicator of the nitrogen source, as the slightly negative values (e.g. 5‰) of plants from natural environments can become very negative (−11.2‰) in sites with agricultural and husbandry activities, but very positive (13.3‰) in urban environments with high vehicular activity. Mosses are good biomonitors for wet deposition and atmospheric epiphytes for dry deposition. In turn, the nitrogen saturation of ecosystems can be monitored with trees whose isotopic values increase with saturation. Although given ecophysiological limitations of different organisms, particular studies should be conducted in each area of interest to determine the most suitable biomonitors. Overall, biomonitors can provide an integrative approach for characterizing nitrogen deposition in regions where the deployment of automated instruments or passive monitoring is not feasible or can be complementary.

This paper investigates intrinsic and extrinsic influences on otariid energetics. We conducted metabolic experiments with three species of Australian otariid and found that sex and water temperature, but not species, significantly influenced metabolic rates. These findings have implications for understanding the physiological capability of otariids to adapt to changing environments., The study of marine mammal energetics can shed light on how these animals might adapt to changing environments. Their physiological potential to adapt will be influenced by extrinsic factors, such as temperature, and by intrinsic factors, such as sex and reproduction. We measured the standard metabolic rate (SMR) of males and females of three Australian otariid species (two Australian fur seals, three New Zealand fur seals and seven Australian sea lions). Mean SMR ranged from 0.47 to 1.05 l O2 min−1, which when adjusted for mass was from 5.33 to 7.44 ml O2 min−1 kg−1. We found that Australian sea lion mass-specific SMR (sSMR; in millilitres of oxygen per minute per kilogram) varied little in response to time of year or moult, but was significantly influenced by sex and water temperature. Likewise, sSMR of Australian and New Zealand fur seals was also influenced by sex and water temperature, but also by time of year (pre-moult, moult or post-moult). During the moult, fur seals had significantly higher sSMR than at other times of the year, whereas there was no discernible effect of moult for sea lions. For both groups, females had higher sSMR than males, but sea lions and fur seals showed different responses to changes in water temperature. The sSMR of fur seals increased with increasing water temperature, whereas sSMR of sea lions decreased with increasing water temperature. There were no species differences when comparing animals of the same sex. Our study suggests that fur seals have more flexibility in their physiology than sea lions, perhaps implying that they will be more resilient in a changing environment.

In this paper we present a new analytical method that allows the extraction of the stress hormone cortisol from naturally sloughed cetacean skin cells. Skin cortisol is unaffected by short-term cortisol fluctuations and could thus be used to assess chronic stress levels in cetaceans., We developed a chemical analytical procedure for sampling, extracting and determining epidermal skin cortisol concentrations (SCCs) in the harbour porpoise (Phocoena phocoena) using gas chromatography–tandem mass spectrometry. In brief, this involved a pressurized liquid extraction with a two-step solid-phase clean-up. A derivatization step was conducted prior to detection. To evaluate the new assay, cortisol was analysed in three different sample types obtained from four harbour porpoises: skin plates, dorsal fin skin plugs (with and without lidocaine) and epidermal scrapes. Skin cortisol concentrations could be measured using the new assay in the majority of the tested skin samples down to a minimal sample size of 49 mg dry weight (dw). Water content ranged from 10 to 46% in the plug samples, which had SCCs from 2.1 to 77.7 ng/g dw. Epidermal scrape samples had the highest water content (83–87%) and lower SCCs (0.6–15 ng/g dw), while the skin plates had intermediate water contents (60–66%) and SCCs of 2.6–13.0 ng/g dw. SCC was slightly higher in plugs with lidocaine than without (average values of 41 and 33 ng/g dw, respectively). Substantial within-individual variations in cortisol concentrations are also common in other matrices such as blood and hair. Some important factors behind this variation could be e.g. the animal's sex, age, body condition, reproductive stage, and the body region sampled, as well as season, moulting cycles and water temperature. Clearly, more research into SCCs is required. The findings described here represent the first critical steps towards using epidermal skin cell samples to assess chronic stress levels in cetaceans and the development of a widely applicable health-assessment tool in these species.

Although not well known, Arctic grayling can move through saline waters and are captured regularly in nearshore coastal waters in Arctic Canada and Alaska with salinities up to 18 ppt. We highlight the implications this has for Blair et al. (2016), a paper recently published in Conservation Physiology.

This review focuses on non-invasive endocrinology, which is a key component of amphibian conservation physiology. It enables rapid assessment of reproductive and stress hormones in free-living and captive populations. It also provides a direct physiological measure of population sensitivity to extreme environments and their sub-lethal impacts on reproduction, health and survival., Non-invasive endocrinology utilizes non-invasive biological samples (such as faeces, urine, hair, aquatic media, and saliva) for the quantification of hormones in wildlife. Urinary-based enzyme immunoassay (EIA) and radio-immunoassay have enabled the rapid quantification of reproductive and stress hormones in amphibians (Anura: Amphibia). With minimal disturbance, these methods can be used to assess the ovarian and testicular endocrine functions as well as physiological stress in captive and free-living populations. Non-invasive endocrine monitoring has therefore greatly advanced our knowledge of the functioning of the stress endocrine system (the hypothalamo–pituitary–interrenal axis) and the reproductive endocrine system (the hypothalamo–pituitary–gonadal axis) in the amphibian physiological stress response, reproductive ecology, health and welfare, and survival. Biological (physiological) validation is necessary for obtaining the excretory lag time of hormone metabolites. Urinary-based EIA for the major reproductive hormones, estradiol and progesterone in females and testosterone in males, can be used to track the reproductive hormone profiles in relationship to reproductive behaviour and environmental data in free-living anurans. Urinary-based corticosterone metabolite EIA can be used to assess the sublethal impacts of biological stressors (such as invasive species and pathogenic diseases) as well as anthropogenic induced environmental stressors (e.g. extreme temperatures) on free-living populations. Non-invasive endocrine methods can also assist in the diagnosis of success or failure of captive breeding programmes by measuring the longitudinal patterns of changes in reproductive hormones and corticosterone within captive anurans and comparing the endocrine profiles with health records and reproductive behaviour. This review paper focuses on the reproductive and the stress endocrinology of anurans and demonstrates the uses of non-invasive endocrinology for advancing amphibian conservation physiology. It also provides key technical considerations for future research that will increase the accuracy and reliability of the data and the value of non-invasive endocrinology within the conceptual framework of conservation physiology.