A body composition model with multiple storage compartments for polar bears (Ursus maritimus).

Climate warming is rapidly altering Arctic ecosystems. Polar bears (Ursus maritimus) need sea ice as a platform from which to hunt seals, but increased sea-ice loss is lengthening periods when bears are without access to primary hunting habitat. During periods of food scarcity, survival depends on the energy that a bear has stored in body reserves, termed storage energy, making this a key metric in predictive models assessing climate change impacts on polar bears. Here, we developed a body composition model for polar bears that estimates storage energy while accounting for changes in storage tissue composition. We used data of dissected polar bears (n = 31) to link routinely collected field measures of total body mass and straight-line body length to the body composition of individual bears, described in terms of structural mass and two storage compartments, adipose and muscle. We then estimated the masses of metabolizable proteins and lipids within these storage compartments, giving total storage energy. We tested this multi-storage model by using it to predict changes in the lipid stores from an independent dataset of wild polar bears (n = 36) that were recaptured 8-200 days later. Using length and mass measurements, our model successfully predicted direct measurements of lipid changes via isotopic dilutions (root mean squared error of 14.5 kg). Separating storage into two compartments, and allowing the molecular composition of storage to vary, provides new avenues for quantifying energy stores of individuals across their life cycle. The multi-storage body composition model thus provides a basis for further exploring energetic costs of physiological processes that contribute to individual survival and reproductive success. Given bioenergetic models are increasingly used as a tool to predict individual fitness and population dynamics, our approach for estimating individual energy stores could be applicable to a wide range of species.

Cortisol levels in blood and hair of unanesthetized grizzly bears (Ursus arctos) following intravenous cosyntropin injection.

Hair cortisol concentration (HCC) is being used increasingly to evaluate long-term stress in many mammalian species. Most of the cortisol is assumed to passively diffuse from circulating blood into hair follicles and gradually accumulate in growing hair. However, our research with free-ranging grizzly bears (Ursus arctos) suggests HCC increases significantly within several hours following capture, a time too brief to be explained by this mechanism alone. In this study with captive grizzly bears, we sought to determine if a brief spike in blood cortisol concentration, thus mimicking a single stressful event, would cause an increase in HCC over a 7-day period. To do this, we administered a single intravenous dose (5 µg/kg) of cosyntropin to three captive unanaesthetised adult female grizzly bears on two occasions, during April when hair growth was arrested and during August when hair was growing. In both trials, the cosyntropin caused a two-fold or greater increase in serum cortisol levels within 1 hr but did not appear to influence HCC at 1, 48, and 168 hr following cosyntropin administration. We conclude the cosyntropin-induced cortisol spike was likely insignificant when compared to the adrenocortical response that occurs in free-ranging bears when captured. We suggest further study with a larger sample of captive bears to evaluate the combined effects of anaesthesia and multiple doses of cosyntropin administered over several hours would better simulate the adrenocortical response of free-ranging grizzly bears during capture.

Updating the AIHTS Trapping Standards to Improve Animal Welfare and Capture Efficiency and Selectivity.

In 1999, after pressure from the European Union, an Agreement on International Humane Trapping Standards (AIHTS) that would result in the banning of the steel-jawed leghold traps in the European Community, Canada, and Russia was signed. The United States implemented these standards through an Agreed Minute with the European Community. Over the last two decades, scientists have criticized the AIHTS for (1) omitting species that are commonly trapped; (2) threshold levels of trap acceptance that are not representative of state-of-the-art trap technology; (3) excluding popular traps which are commonly used by trappers although they are known to cause prolonged pain and stress to captured animals; (4) inadequate coverage of capture efficiency and species selectivity (i.e., number of captures of target and non-target species) performance. Concerns about the ability of standards and test procedures to ensure animal welfare, and about the implementation of standards, have also been voiced by wildlife biologists, managers, and conservation groups. In this review, we present a synopsis of current trapping standards and test procedures, and we compare the standards to a then contemporary 1985-1993 Canadian trap research and development program. On the basis of the above-noted concerns about AIHTS, and our experience as wildlife professionals involved in the capture of mammals, we formulated the following hypotheses: (1) the list of mammal species included in the AIHTS is incomplete; (2) the AIHTS have relatively low animal welfare performance thresholds of killing trap acceptance and do not reflect state-of-the-art trapping technology; (3) the AIHTS animal welfare indicators and injuries for restraining traps are insufficient; (4) the AIHTS testing procedures are neither thorough nor transparent; (5) the AIHTS protocols for the use of certified traps are inadequate; (6) the AIHTS procedures for the handling and dispatching of animals are nonexistent; (7) the AIHTS criteria to assess trap capture efficiency and species selectivity are inappropriate. We conclude that the AIHTS do not reflect state-of-the-art trapping technology, and assessment protocols need to be updated to include trap components and sets, animal handling and dispatching, and trap visit intervals. The list of traps and species included in the standards should be updated. Finally, the concepts of capture efficiency and trap selectivity should be developed and included in the standards. Based on our review, it is clear that mammal trapping standards need to be revisited to implement state-of-the-art trapping technology and improve capture efficiency and species selectivity. We believe that a committee of international professionals consisting of wildlife biologists and veterinarians with extensive experience in the capture of mammals and animal welfare could produce new standards within 1-2 years. We propose a series of measures to fund trap testing and implement new standards.

Hair Cortisol Concentration and Body Mass in Moose (Alces alces) Infested with Deer Keds (Lipoptena cervi).

The deer ked (Lipoptena cervi), a hematophagous ectoparasite of cervids, is currently spreading in Scandinavia, and the moose (Alces alces) is its main host. However, little is known about the impact of deer keds on moose. We analyzed the hair cortisol concentration (HCC) from 262 moose harvested in the fall in relation to age class, sex, body mass (BM), and deer ked infestation intensity, and BM in relation to age class, sex, and infestation intensity. We found that HCC decreased with increasing deer ked intensity at low ked intensities, but for the higher levels of ked intensities, there was a positive relationship between HCC and ked intensity. The HCC was higher in males than in females and lower in yearlings than in calves and adults. Our failure to find any association between BM and deer ked intensity suggested a negligible impact of deer ked infestation on moose foraging and metabolism at the level of infestation observed early in the infestation, but did not exclude an effect later in winter. Our findings suggested that moose generally tolerated moderate parasitism by keds. However, the increase in HCC at higher ked intensities suggested that the tolerance strategy could be disrupted with further increases in intensities and consequently may negatively affect animal health and welfare.

Do follicles matter? Testing the effect of follicles on hair cortisol levels.

Cortisol concentrations in hair are used increasingly as a biomarker of long-term stress in free-ranging wildlife. Cortisol is believed to be integrated into hair primarily during its active growth phase, typically occurring over weeks to months or longer periods, depending on latitude. Cortisol concentrations in hair thus reflect the activity of the hypothalamic-pituitary-adrenal axis over this time. However, local, independent cortisol secretion within the skin, which includes hair follicles, may also contribute to cortisol levels in growing hair. Methodological differences between studies include the measurement of cortisol in only the hair shaft (i.e. follicle absent, as with shaved hair) versus the whole hair (i.e. follicle present, as with plucked hair). If the concentration of cortisol in the follicle is high enough to influence the overall hair cortisol concentration (HCC), this could confound comparisons between studies using different types of hair samples (hair shafts vs. whole hair) and collection methods. Here, we test the hypothesis that cortisol present in follicles influences HCC. We compared HCC in paired subsamples of hair with and without follicles from 30 free-ranging Scandinavian brown bears (Ursus arctos) and observed significantly greater HCC in samples with follicles present. The effect of follicles remained significant also with sex and age of sampled bears taken into account in a linear mixed model. Finally, we provide an overview of collection methods and types of hair samples used for HCC analysis in 77 studies dealing with stress in wild mammal species. Our findings highlight the need to unify methods of hair collection and preparation to allow for valid comparisons, and to optimize labour input in ecophysiological studies.

Environment, endocrinology, and biochemistry influence expression of stress proteins in bottlenose dolphins.

Natural and anthropogenic stressors have been reported to impact the health of marine mammals. Therefore, investigation of quantifiable biomarkers in response to stressors is required. We hypothesized that stress protein expression would be associated with biological and health variables in wild and managed-care bottlenose dolphins (Tursiops truncatus). To test this hypothesis, our study objectives were to (1) determine if stress proteins in skin, white blood cells (WBCs), and plasma could be measured with an antibody-based microarray, (2) measure stress-protein expression relative to biological data (location, sex, age, environment), and (3) determine if stress-protein expression was associated with endocrine, hematological, biochemical and serological variables and gene expression in bottlenose dolphins. Samples were collected from two wild groups (n = 28) and two managed-care groups (n = 17). Proteins involved in the HPA axis, apoptosis, proteotoxicity, and inflammation were identified as stress proteins. The expression of 3 out of 33 proteins was significantly (P < 0.05) greater in skin than plasma and WBCs. Male dolphins had significantly greater expression levels for 10 proteins in skin compared to females. The greatest number of stress-associated proteins varied by the dolphins' environment; nine were greater in managed-care dolphins and 15 were greater in wild dolphins, which may be related to wild dolphin disease status. Protein expression in skin and WBCs showed many positive relationships with measures of plasma endocrinology and biochemistry. This study provides further understanding of the underlying mechanisms of the stress response in bottlenose dolphins and application of a combination of novel methods to measure stress in wildlife.

Can concentrations of steroid hormones in brown bear hair reveal age class?

Although combining genetic and endocrine data from non-invasively collected hair samples has potential to improve the conservation of threatened mammals, few studies have evaluated this opportunity. In this study, we determined if steroid hormone (testosterone, progesterone, estradiol and cortisol) concentration profiles in 169 hair samples collected from free-ranging brown bears (Ursus arctos) could be used to accurately discriminate between immature and adult bears within each sex. Because hair samples were acquired opportunistically, we also needed to establish if interactions between hormones and several non-hormone factors (ordinal day, year, contact method, study area) were associated with age class. For each sex, we first compared a suite of candidate models by Akaike Information Criteria model selection, using different adult-age thresholds (3, 4 and 5 years), to determine the most supported adult age. Because hair hormone levels better reflect the endocrine state at an earlier time, possibly during the previous year, then at the time of sampling, we re-analysed the data, excluding the records for bears at the adult-age threshold, to establish if classification accuracy improved. For both sexes, candidate models were most supported based on a 3-year-old adult-age threshold. Classification accuracy did not improve with the 3-year-old bear data excluded. Male age class was predicted with a high degree of accuracy (88.4%) based on the concomitant concentrations of all four hormones. Female age class was predicted with less accuracy (77.1%) based only on testosterone and cortisol. Accuracy was reduced for females, primarily because we had poor success in correctly classifying immature bears (60%) whereas classification success for adult females was similar to that for males (84.5%). Given the small and unbalanced sample used in this study, our findings should be viewed as preliminary, but they should also provide a basis for more comprehensive future studies.

Compatibility of preparatory procedures for the analysis of cortisol concentrations and stable isotope (δ13C, δ15N) ratios: a test on brown bear hair.

The measurement of naturally occurring glucocorticoids and stable isotopes of several elements has gained importance in wildlife studies in recent decades and opened a myriad of ecological applications. Cortisol and stable isotopes equilibrate in animal tissues over periods of integration related to the growth rate of the tissue, providing information reflecting systemic cortisol secretion and dietary intake. Sample preparation shares the common step of first cleaning the sample of external contamination. However, it is not well understood how different solvents used in sample preparation affect isotopic and cortisol values, and whether it is safe to follow the same procedures for both measures to optimize analyses of the same sample. We conducted an experiment to compare different preparation protocols for the analysis of cortisol concentrations and stable carbon (δ13C) and nitrogen (δ15N) isotope ratios in hair. Hair samples from 12 brown bears (Ursus arctos) were each divided into five aliquots; two aliquots were rinsed with a 2:1 chloroform:methanol (v/v) mixture with one aliquot ground prior to cortisol analysis and the other left intact for stable isotope analyses; two aliquots were washed with methanol with one aliquot ground prior to cortisol analysis and the other left intact for stable isotope analyses; and one aliquot washed with methanol and ground prior to stable isotope analyses. The cortisol, δ13C and δ15N values remained consistent following all treatments. Our results indicate that hair samples rinsed with a 2:1 chloroform:methanol mixture or washed with methanol can be used for both types of analyses. Further, hair that has been ground in a standard hair cortisol procedure can also be used for stable isotope analysis. This information is important for improving laboratory efficiency and compatibility of procedures used for wildlife physiological ecology studies where concurrent measurements of cortisol and stable isotopes in hair are required.

The quantification of reproductive hormones in the hair of captive adult brown bears and their application as indicators of sex and reproductive state.

Recognizing the potential value of steroid hormone measurements to augment non-invasive genetic sampling, we developed procedures based on enzyme-linked immunoassays to quantify reproductive steroid hormone concentrations in brown bear (Ursus arctos) hair. Then, using 94 hair samples collected from eight captive adult bears over a 2-year period, we evaluated (i) associations between hair concentrations of testosterone, progesterone, estradiol and cortisol; (ii) the effect of collecting by shaving vs. plucking; and (iii) the utility of reproductive hormone profiles to differentiate sex and reproductive state. Sample requirements (125 mg of guard hair) to assay all hormones exceeded amounts typically obtained by non-invasive sampling. Thus, broad application of this approach will require modification of non-invasive techniques to collect larger samples, use of mixed (guard and undercoat) hair samples and/or application of more sensitive laboratory procedures. Concentrations of hormones were highly correlated suggesting their sequestration in hair reflects underlying physiological processes. Marked changes in hair hormone levels during the quiescent phase of the hair cycle, coupled with the finding that progesterone concentrations, and their association with testosterone levels, differed markedly between plucked and shaved hair samples, suggests steroids sequestered in hair were likely derived from various sources, including skin. Changes in hair hormone concentrations over time, and in conjunction with key reproductive events, were similar to what has been reported concerning hormonal changes in the blood serum of brown bears. Thus, potential for the measurement of hair reproductive hormone levels to augment non-invasive genetic sampling appears compelling. Nonetheless, we are conducting additional validation studies on hair collected from free-ranging bears, representative of all sex, age and reproductive classes, to fully evaluate the utility of this approach for brown bear conservation and research.

Comparison of methanol and isopropanol as wash solvents for determination of hair cortisol concentration in grizzly bears and polar bears.

Methodological differences among laboratories are recognized as significant sources of variation in quantification of hair cortisol concentration (HCC). An important step in processing hair, particularly when collected from wildlife, is the choice of solvent used to remove or "wash" external hair shaft cortisol prior to quantification of HCC. The present study systematically compared methanol and isopropanol as wash solvents for their efficiency at removing external cortisol without extracting internal hair shaft cortisol in samples collected from free-ranging grizzly bears and polar bears. Cortisol concentrations in solvents and hair were determined in each of one to eight washes of hair with each solvent independently. •There were no significant decreases in internal hair shaft cortisol among all eight washes for either solvent, although methanol removed detectable hair surface cortisol after one wash in grizzly bear hair whereas hair surface cortisol was detected in all eight isopropanol washes.•There were no significant differences in polar bear HCC washed one to eight times with either solvent, but grizzly bear HCC was significantly greater in hair washed with isopropanol compared to methanol.•There were significant differences in HCC quantified using different commercial ELISA kits commonly used for HCC determinations.

A Double-Blinded, Randomized Comparison of Medetomidine-Tiletamine-Zolazepam and Dexmedetomidine-Tiletamine-Zolazepam Anesthesia in Free-Ranging Brown Bears (Ursus Arctos).

We compared anesthetic features, blood parameters, and physiological responses to either medetomidine-tiletamine-zolazepam or dexmedetomidine-tiletamine-zolazepam using a double-blinded, randomized experimental design during 40 anesthetic events of free-ranging brown bears (Ursus arctos) either captured by helicopter in Sweden or by culvert trap in Canada. Induction was smooth and predictable with both anesthetic protocols. Induction time, the need for supplemental drugs to sustain anesthesia, and capture-related stress were analyzed using generalized linear models, but anesthetic protocol did not differentially affect these variables. Arterial blood gases and acid-base status, and physiological responses were examined using linear mixed models. We documented acidemia (pH of arterial blood < 7.35), hypoxemia (partial pressure of arterial oxygen < 80 mmHg), and hypercapnia (partial pressure of arterial carbon dioxide ≥ 45 mmHg) with both protocols. Arterial pH and oxygen partial pressure were similar between groups with the latter improving markedly after oxygen supplementation (p < 0.001). We documented dose-dependent effects of both anesthetic protocols on induction time and arterial oxygen partial pressure. The partial pressure of arterial carbon dioxide increased as respiratory rate increased with medetomidine-tiletamine-zolazepam, but not with dexmedetomidine-tiletamine-zolazepam, demonstrating a differential drug effect. Differences in heart rate, respiratory rate, and rectal temperature among bears could not be attributed to the anesthetic protocol. Heart rate increased with increasing rectal temperature (p < 0.001) and ordinal day of capture (p = 0.002). Respiratory rate was significantly higher in bears captured by helicopter in Sweden than in bears captured by culvert trap in Canada (p < 0.001). Rectal temperature significantly decreased over time (p ≤ 0.05). Overall, we did not find any benefit of using dexmedetomidine-tiletamine-zolazepam instead of medetomidine-tiletamine-zolazepam in the anesthesia of brown bears. Both drug combinations appeared to be safe and reliable for the anesthesia of free-ranging brown bears captured by helicopter or by culvert trap.

Development and application of an antibody-based protein microarray to assess physiological stress in grizzly bears (Ursus arctos).

A novel antibody-based protein microarray was developed that simultaneously determines expression of 31 stress-associated proteins in skin samples collected from free-ranging grizzly bears (Ursus arctos) in Alberta, Canada. The microarray determines proteins belonging to four broad functional categories associated with stress physiology: hypothalamic-pituitary-adrenal axis proteins, apoptosis/cell cycle proteins, cellular stress/proteotoxicity proteins and oxidative stress/inflammation proteins. Small skin samples (50-100 mg) were collected from captured bears using biopsy punches. Proteins were isolated and labelled with fluorescent dyes, with labelled protein homogenates loaded onto microarrays to hybridize with antibodies. Relative protein expression was determined by comparison with a pooled standard skin sample. The assay was sensitive, requiring 80 µg of protein per sample to be run in triplicate on the microarray. Intra-array and inter-array coefficients of variation for individual proteins were generally <10 and <15%, respectively. With one exception, there were no significant differences in protein expression among skin samples collected from the neck, forelimb, hindlimb and ear in a subsample of n = 4 bears. This suggests that remotely delivered biopsy darts could be used in future sampling. Using generalized linear mixed models, certain proteins within each functional category demonstrated altered expression with respect to differences in year, season, geographical sampling location within Alberta and bear biological parameters, suggesting that these general variables may influence expression of specific proteins in the microarray. Our goal is to apply the protein microarray as a conservation physiology tool that can detect, evaluate and monitor physiological stress in grizzly bears and other species at risk over time in response to environmental change.

LEUKOCYTE COPING CAPACITY AS A TOOL TO ASSESS CAPTURE- AND HANDLING-INDUCED STRESS IN SCANDINAVIAN BROWN BEARS (URSUS ARCTOS).

Brown bears (Ursus arctos) are often captured and handled for research and management purposes. Although the techniques used are potentially stressful for the animals and might have detrimental and long-lasting consequences, it is difficult to assess their physiological impact. Here we report the use of the leukocyte coping capacity (LCC) technique to quantify the acute stress of capture and handling in brown bears in Scandinavia. In April and May 2012 and 2013, we collected venous blood samples and recorded a range of physiological variables to evaluate the effects of capture and the added impact of surgical implantation or removal of transmitters and sensors. We studied 24 brown bears, including 19 that had abdominal surgery. We found 1) LCC values following capture were lower in solitary bears than in bears in family groups suggesting capture caused relatively more stress in solitary bears, 2) ability to cope with handling stress was better (greater LCC values) in bears with good body condition, and 3) LCC values did not appear to be influenced by surgery. Although further evaluation of this technique is required, our preliminary results support the use of the LCC technique as a quantitative measure of stress.

Quantifying long-term stress in brown bears with the hair cortisol concentration: a biomarker that may be confounded by rapid changes in response to capture and handling.

The measurement of cortisol in hair is becoming important in studying the role of stress in the life history, health and ecology of wild mammals. The hair cortisol concentration (HCC) is generally believed to be a reliable indicator of long-term stress that can reflect frequent or prolonged activation of the hypothalamic-pituitary-adrenal axis over weeks to months through passive diffusion from the blood supply to the follicular cells that produce the hair. Diffusion of cortisol from tissues surrounding the follicle and glandular secretions (sebum and sweat) that coat the growing hair may also affect the HCC, but the extent of these effects is thought to be minimal. In this study, we report on a range of factors that are associated with, and possibly influence, cortisol concentrations in the hair of free-ranging brown bears (Ursus arctos). Through two levels of analyses that differed in sample sizes and availability of predictor variables, we identified the presence or absence of capture, restraint and handling, as well as different methods of capture, as significant factors that appeared to influence HCC in a time frame that was too short (minutes to hours) to be explained by passive diffusion from the blood supply alone. Furthermore, our results suggest that HCC was altered after hair growth had ceased and blood supply to the hair follicle was terminated. However, we also confirmed that HCC was inversely associated with brown bear body condition and was, therefore, responsive to diminished food availability/quality and possibly other long-term stressors that affect body condition. Collectively, our findings emphasize the importance of further elucidating the mechanisms of cortisol accumulation in hair and the influence of long- and short-term stressors on these mechanisms.

Environmental factors and habitat use influence body condition of individuals in a species at risk, the grizzly bear.

Metrics used to quantify the condition or physiological states of individuals provide proactive mechanisms for understanding population dynamics in the context of environmental factors. Our study examined how anthropogenic disturbance, habitat characteristics and hair cortisol concentrations interpreted as a sex-specific indicator of potential habitat net-energy demand affect the body condition of grizzly bears (n = 163) in a threatened population in Alberta, Canada. We quantified environmental variables by modelling spatial patterns of individual habitat use based on global positioning system telemetry data. After controlling for gender, age and capture effects, we assessed the influence of biological and environmental variables on body condition using linear mixed-effects models in an information theoretical approach. Our strongest model suggested that body condition was improved when patterns of habitat use included greater vegetation productivity, increased influence of forest harvest blocks and oil and gas well sites, and a higher percentage of regenerating and coniferous forest. However, body condition was negatively affected by habitat use in close proximity to roads and in areas where potential energetic demands were high. Poor body condition was also associated with increased selection of parks and protected areas and greater seasonal vegetation productivity. Adult females, females with cubs-of-year, juvenile females and juvenile males were in poorer body condition compared with adult males, suggesting that intra-specific competition and differences in habitat use based on gender and age may influence body condition dynamics. Habitat net-energy demand also tended to be higher in areas used by females which, combined with observed trends in body condition, could affect reproductive success in this threatened population. Our results highlight the importance of considering spatiotemporal variability in environmental factors and habitat use when assessing the body condition of individuals. Long-term and large-scale monitoring of the physiological state of individuals provides a more comprehensive approach to support management and conservation of species at risk.

Environmental, biological and anthropogenic effects on grizzly bear body size: temporal and spatial considerations.

BACKGROUND: Individual body growth is controlled in large part by the spatial and temporal heterogeneity of, and competition for, resources. Grizzly bears (Ursus arctos L.) are an excellent species for studying the effects of resource heterogeneity and maternal effects (i.e. silver spoon) on life history traits such as body size because their habitats are highly variable in space and time. Here, we evaluated influences on body size of grizzly bears in Alberta, Canada by testing six factors that accounted for spatial and temporal heterogeneity in environments during maternal, natal and 'capture' (recent) environments. After accounting for intrinsic biological factors (age, sex), we examined how body size, measured in mass, length and body condition, was influenced by: (a) population density; (b) regional habitat productivity; (c) inter-annual variability in productivity (including silver spoon effects); (d) local habitat quality; (e) human footprint (disturbances); and (f) landscape change. RESULTS: We found sex and age explained the most variance in body mass, condition and length (R(2) from 0.48-0.64). Inter-annual variability in climate the year before and of birth (silver spoon effects) had detectable effects on the three-body size metrics (R(2) from 0.04-0.07); both maternal (year before birth) and natal (year of birth) effects of precipitation and temperature were related with body size. Local heterogeneity in habitat quality also explained variance in body mass and condition (R(2) from 0.01-0.08), while annual rate of landscape change explained additional variance in body length (R(2) of 0.03). Human footprint and population density had no observed effect on body size. CONCLUSIONS: These results illustrated that body size patterns of grizzly bears, while largely affected by basic biological characteristics (age and sex), were also influenced by regional environmental gradients the year before, and of, the individual's birth thus illustrating silver spoon effects. The magnitude of the silver spoon effects was on par with the influence of contemporary regional habitat productivity, which showed that both temporal and spatial influences explain in part body size patterns in grizzly bears. Because smaller bears were found in colder and less-productive environments, we hypothesize that warming global temperatures may positively affect body mass of interior bears.

Falling through the cracks: shortcomings in the collaboration between biologists and veterinarians and their consequences for wildlife.

Although biologists and veterinarians have shown considerable success in working together to address wildlife-related issues, including disease, chemical immobilization, reproductive biology, and conservation biology, examples of shared efforts to evaluate and ensure the welfare of study animals are mostly absent. I present the case that this deficiency arises primarily from a lack of mutual understanding between fields with respect to the other's training and experience in addressing animal welfare issues. In effect, each assumes that the final word on animal welfare rests with the other. The reality is, however, that neither field contains the knowledge and skills required to address animal welfare concerns alone. Nevertheless, wildlife researchers are increasingly encountering difficulties conducting research on wild animals because of opposition from stakeholders on the basis of animal welfare concerns. Further, a growing number of articles in the peer-reviewed scientific literature are reporting on potential biases in research results developing from the welfare impacts of widely used techniques, including methods of capturing and marking wildlife. By viewing animal welfare as a shared responsibility and combining their knowledge and skills, wildlife biologists and veterinarians have an opportunity to reform "invasive" wildlife research in a manner that is less harmful to the animals being studied, less likely to bias research results, and less objectionable to the stakeholders who ultimately influence or make decisions on how wildlife research is conducted.

Spatial analysis of factors influencing long-term stress in the grizzly bear (Ursus arctos) population of Alberta, Canada.

Non-invasive measures for assessing long-term stress in free ranging mammals are an increasingly important approach for understanding physiological responses to landscape conditions. Using a spatially and temporally expansive dataset of hair cortisol concentrations (HCC) generated from a threatened grizzly bear (Ursus arctos) population in Alberta, Canada, we quantified how variables representing habitat conditions and anthropogenic disturbance impact long-term stress in grizzly bears. We characterized spatial variability in male and female HCC point data using kernel density estimation and quantified variable influence on spatial patterns of male and female HCC stress surfaces using random forests. Separate models were developed for regions inside and outside of parks and protected areas to account for substantial differences in anthropogenic activity and disturbance within the study area. Variance explained in the random forest models ranged from 55.34% to 74.96% for males and 58.15% to 68.46% for females. Predicted HCC levels were higher for females compared to males. Generally, high spatially continuous female HCC levels were associated with parks and protected areas while low-to-moderate levels were associated with increased anthropogenic disturbance. In contrast, male HCC levels were low in parks and protected areas and low-to-moderate in areas with increased anthropogenic disturbance. Spatial variability in gender-specific HCC levels reveal that the type and intensity of external stressors are not uniform across the landscape and that male and female grizzly bears may be exposed to, or perceive, potential stressors differently. We suggest observed spatial patterns of long-term stress may be the result of the availability and distribution of foods related to disturbance features, potential sexual segregation in available habitat selection, and may not be influenced by sources of mortality which represent acute traumas. In this wildlife system and others, conservation and management efforts can benefit by understanding spatial- and gender-based stress responses to landscape conditions.

Serum corticosteroid binding globulin expression is modulated by fasting in polar bears (Ursus maritimus).

Polar bears (Ursus maritimus) from several subpopulations undergo extended fasting during the ice-free season. However, the animals appear to conserve protein despite the prolonged fasting, though the mechanisms involved are poorly understood. We hypothesized that elevated concentrations of corticosteroid binding globulin (CBG), the primary cortisol binding protein in circulation, lead to cortisol resistance and provide a mechanism for protein conservation during extended fasting. The metabolic state (feeding vs. fasting) of 16 field sampled male polar bears was determined based on their serum urea to creatinine ratio (>25 for feeding vs. <5 for fasting). There were no significant differences in serum cortisol levels between all male and female polar bears sampled. Serum CBG expression was greater in lactating females relative to non-lactating females and males. CBG expression was significantly higher in fasting males when compared to non-fasting males. This leads us to suggest that CBG expression may serve as a mechanism to conserve protein during extended fasting in polar bears by reducing systemic free cortisol concentrations. This was further supported by a lower serum glucose concentration in the fasting bears. As well, a lack of an enhanced adrenocortical response to acute capture stress supports our hypothesis that chronic hunger is not a stressor in this species. Overall, our results suggest that elevated serum CBG expression may be an important adaptation to spare proteins by limiting cortisol bioavailability during extended fasting in polar bears.