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.

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.

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.

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.

Grizzly bear corticosteroid binding globulin: Cloning and serum protein expression.

Serum corticosteroid levels are routinely measured as markers of stress in wild animals. However, corticosteroid levels rise rapidly in response to the acute stress of capture and restraint for sampling, limiting its use as an indicator of chronic stress. We hypothesized that serum corticosteroid binding globulin (CBG), the primary transport protein for corticosteroids in circulation, may be a better marker of the stress status prior to capture in grizzly bears (Ursus arctos). To test this, a full-length CBG cDNA was cloned and sequenced from grizzly bear testis and polyclonal antibodies were generated for detection of this protein in bear sera. The deduced nucleotide and protein sequences were 1218 bp and 405 amino acids, respectively. Multiple sequence alignments showed that grizzly bear CBG (gbCBG) was 90% and 83% identical to the dog CBG nucleotide and amino acid sequences, respectively. The affinity purified rabbit gbCBG antiserum detected grizzly bear but not human CBG. There were no sex differences in serum total cortisol concentration, while CBG expression was significantly higher in adult females compared to males. Serum cortisol levels were significantly higher in bears captured by leg-hold snare compared to those captured by remote drug delivery from helicopter. However, serum CBG expression between these two groups did not differ significantly. Overall, serum CBG levels may be a better marker of chronic stress, especially because this protein is not modulated by the stress of capture and restraint in grizzly bears.

Use of hyaluronidase to improve chemical immobilization of free-ranging polar bears (Ursus maritimus).

We assessed the efficacy and safety of hyaluronidase to improve chemical immobilization of free-ranging polar bears (Ursus maritimus) captured from helicopter by remote drug delivery along the Ontario coast line of northwestern James Bay and southern Hudson Bay during September 2005 and October 2007. We used a single blind study design in which one person prepared and loaded all darts without the shooter knowing whether hyaluronidase (150 IU per dart) or sterile water was added to the immobilizing drug mixture of xylazine and zolazepam-tiletamine (XZT). We found that we often required more than one dart to immobilize bears in the control group (XZT+sterile water; >1 dart for 15 of 28 captures) versus the treatment group (XZT+hyaluronidase; >1 dart for seven of 26 captures). As a consequence, treatment bears were generally immobilized with smaller XZT dosages (7.9 vs. 9.4 mg/kg; P = 0.08) and shorter induction (10 vs. 15 min; P = 0.004) than control bears. We found no differences in vital rates and serum biochemistry results between control and treatment bears. We did find, however, that induction times correlated directly with rectal temperature at

Intranasal administration of xylazine to reduce stress in elk captured by net gun.

Forty free-ranging elk (Cervus elaphus manitobensis) were captured by net gun in Riding Mountain National Park (Manitoba, Canada) during February 2002 and were administered either saline (control) or xylazine by the intranasal route, to evaluate the efficacy and benefit of intranasal xylazine to reduce stress. Elk that received xylazine had higher relaxation scores than control elk, and the onset of sedation occurred quickly, often <1 min. Serum concentrations of cortisol, creatine kinase, and gamma-glutamyltransferase were lower in elk that received xylazine than in control elk. At the conclusion of handling, the intravenous administration of yohimbine quickly abolished the sedative effect of xylazine, which allowed elk to be released without concern of physical injury due to ataxia. The intranasal administration of xylazine can be used to reduce stress in wild animals under situations where they are being handled while physically restrained.

Antibodies to canine distemper and phocine distemper viruses in polar bears from the Canadian arctic.

Serum samples collected from 200 polar bears (Ursus marititnus) from two populations in the Canadian arctic, the western Hudson Bay and Lancaster Sound populations, between 1989 and 1996, were tested for antibodies to canine distemper (CDV) and phocine distemper viruses (PDV) using virus neutralization. Antibodies to CDV and PDV were detected in 48 and six polar bears, respectively. All six bears that tested positive for PDV also tested positive for CDV; in only one case did the antibody titer for PDV exceed that of CDV. Differences in antibody prevalence to CDV were detected between populations and age classes but not sex or year of sampling.