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Hibernation in brown bears is an annual process involving multiple physiologically distinct seasons—hibernation, active, and hyperphagia. While recent studies have characterized broad patterns of differential gene regulation and isoform usage between hibernation and active seasons, patterns of gene and isoform expression during hyperphagia remain relatively poorly understood. The hyperphagia stage occurs between active and hibernation seasons and involves the accumulation of large fat reserves in preparation for hibernation. Here, we use time-series analyses of gene expression and isoform usage to interrogate transcriptomic regulation associated with all three seasons. We identify a large number of genes with significant differential isoform usage (DIU) across seasons and show that these patterns of isoform usage are largely tissue-specific. We also show that DIU and differential gene-level expression responses are generally non-overlapping, with only a small subset of multi-isoform genes showing evidence of both gene-level expression changes and changes in isoform usage across seasons. Additionally, we investigate nuanced regulation of candidate genes involved in the insulin signaling pathway and find evidence of hyperphagia-specific gene expression and isoform regulation that may enhance fat accumulation during hyperphagia. Our findings highlight the value of using temporal analyses of both gene- and isoform-level gene expression when interrogating complex physiological phenotypes and provide new insight into the mechanisms underlying seasonal changes in bear physiology.

Although polar bears (Ursus maritimus) and brown bears (U. arctos) have been exhibited in zoological gardens for centuries, little is known about their nutritional needs. Multiple recent studies on both wild and captive polar bears and brown bears have found that they voluntarily select dietary macronutrient proportions resulting in much lower dietary protein and higher fat or digestible carbohydrate concentrations than are currently fed in most zoos. These lower protein concentrations selected by both species maximized growth rates and efficiencies of energy utilization in brown bears and may play a role in reducing kidney, liver, and cardiovascular diseases in both species. Therefore, we propose the need for the development of new dietary regimens for both species in managed care that better reflect their macronutrient needs. We developed a new kibble that is higher in fat and lower in protein than typical diets that have been fed in managed care, has a fatty acid profile more consistent with wild bear diets, and has been readily consumed by both brown bears and polar bears. The kibble can be fed as the sole diet or as part of more complex diets with additional fruits, meats, or vegetables. Because many nutritional deficiencies and related diseases can take months or years to appear, we urge caution and continued long-term monitoring of bears and their diets to ensure their optimal health.

Accurate information on diet composition is central to understanding and conserving carnivore populations. Quantitative fatty acid signature analysis (QFASA) has emerged as a powerful tool for estimating the diets of predators, but ambiguities remain about the timeframe of QFASA estimates and the need to account for species-specific patterns of metabolism. We conducted a series of feeding experiments with four juvenile male brown bears (Ursus arctos) to (1) track the timing of changes in adipose tissue composition and QFASA diet estimates in response to a change in diet and (2) quantify the relationship between consumer and diet FA composition (i.e., determine "calibration coefficients"). Bears were fed three compositionally distinct diets for 90-120 days each. Two marine-based diets were intended to approximate the lipid content and composition of the wild diet of polar bears (U. maritimus). Bear adipose tissue composition changed quickly in the direction of the diet and showed evidence of stabilization after 60 days. During hibernation, FA profiles were initially stable but diet estimates after 10 weeks were sensitive to calibration coefficients. Calibration coefficients derived from the marine-based diets were broadly similar to each other and to published values from marine-fed mink (Mustela vison), which have been used as a model for free-ranging polar bears. For growing bears on a high-fat diet, the temporal window for QFASA estimates was 30-90 days. Although our results reinforce the importance of accurate calibration, the similarities across taxa and diets suggest it may be feasible to develop a generalized QFASA approach for mammalian carnivores.

A male mutation bias is observed across vertebrates, and, where data are available, this bias is accompanied by increased per-generation mutation rates with parental age. While continuing mitotic cell division in the male germline post puberty has been proposed as the major cellular mechanism underlying both patterns, little direct evidence for this role has been found. Understanding the evolution of the per-generation mutation rate among species requires that we identify the molecular mechanisms that change between species. Here, we study the per-generation mutation rate in an extended pedigree of the brown (grizzly) bear, Ursus arctos horribilis. Brown bears hibernate for one-third of the year, a period during which spermatogenesis slows or stops altogether. The reduction of spermatogenesis is predicted to lessen the male mutation bias and to lower the per-generation mutation rate in this species. However, using whole-genome sequencing, we find that both male bias and per-generation mutation rates are highly similar to that expected for a non-hibernating species. We also carry out a phylogenetic comparison of substitution rates along the lineage leading to brown bear and panda (a non-hibernating species) and find no slowing of the substitution rate in the hibernator. Our results contribute to accumulating evidence that suggests that male germline cell division is not the major determinant of mutation rates and mutation biases. The results also provide a quantitative basis for improved estimates of the timing of carnivore evolution.

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., Administration of cosyntropin to captive grizzly bears caused increased serum cortisol levels within 1 hr. However, cosyntropin did not influence hair cortisol concentration at 1, 48, and 168 hr following administration. Cosyntropin‐induced hair cortisol levels were insignificant compared to free‐ranging bears during capture.

A male mutation bias is observed across vertebrates, and, where data are available, this bias is accompanied by increased per-generation mutation rates with parental age. While continuing mitotic cell division in the male germline post-puberty has been proposed as the major cellular mechanism underlying both patterns, little direct evidence for this role has been found. Understanding the evolution of the per-generation mutation rate among species requires that we identify the molecular mechanisms that change between species. Here, we study the per-generation mutation rate in an extended pedigree of the brown (grizzly) bear, Ursus arctos horribilis. Brown bears hibernate for one-third of the year, a period during which spermatogenesis slows or stops altogether. The cessation of spermatogenesis is predicted to lessen the male mutation bias and to lower the per-generation mutation rate in this species. However, using whole-genome sequencing, we find that both male bias and per-generation mutation rates are the same as expected for a non-hibernating species. We also carry out a phylogenetic comparison of substitution rates along the lineage leading to brown bear and panda (a non-hibernating species) and find no slowing of the substitution rate in the hibernator. Our results contribute to accumulating evidence that suggests that male germline cell division is not the major determinant of mutation rates and mutation biases. The results also provide a quantitative basis for improved estimates of the timing of carnivore evolution.

Life in the Arctic presents organisms with multiple challenges, including extreme photic conditions, cold temperatures, and annual loss and daily movement of sea ice. Polar bears ( Ursus maritimus) evolved under these unique conditions, where they rely on ice to hunt their main prey, seals. However, very little is known about the dynamics of their daily and seasonal activity patterns. For many organisms, activity is synchronized (entrained) to the earth’s day/night cycle, in part via an endogenous (circadian) timekeeping mechanism. The present study used collar-mounted accelerometer and global positioning system data from 122 female polar bears in the Chukchi and Southern Beaufort Seas collected over an 8-year period to characterize activity patterns over the calendar year and to determine if circadian rhythms are expressed under the constant conditions found in the Arctic. We reveal that the majority of polar bears (80%) exhibited rhythmic activity for the duration of their recordings. Collectively within the rhythmic bear cohort, circadian rhythms were detected during periods of constant daylight (June-August; 24.40 ± 1.39 h, mean ± SD) and constant darkness (23.89 ± 1.72 h). Exclusive of denning periods (November-April), the time of peak activity remained relatively stable (acrophases: ~1200-1400 h) for most of the year, suggesting either entrainment or masking. However, activity patterns shifted during the spring feeding and seal pupping season, as evidenced by an acrophase inversion to ~2400 h in April, followed by highly variable timing of activity across bears in May. Intriguingly, despite the dynamic environmental photoperiodic conditions, unpredictable daily timing of prey availability, and high between-animal variability, the average duration of activity (alpha) remained stable (11.2 ± 2.9 h) for most of the year. Together, these results reveal a high degree of behavioral plasticity in polar bears while also retaining circadian rhythmicity. Whether this degree of plasticity will benefit polar bears faced with a loss of sea ice remains to be determined.

Understanding how metabolic reprogramming happens in cells will aid the progress in the treatment of a variety of metabolic disorders. Brown bears undergo seasonal shifts in insulin sensitivity, including reversible insulin resistance in hibernation. We performed RNA-sequencing on brown bear adipocytes and proteomics on serum to identify changes possibly responsible for reversible insulin resistance. We observed dramatic transcriptional changes, which depended on both the cell and serum season of origin. Despite large changes in adipocyte gene expression, only changes in eight circulating proteins were identified as related to the seasonal shifts in insulin sensitivity, including some that have not previously been associated with glucose homeostasis. The identified serum proteins may be sufficient for shifting hibernation adipocytes to an active-like state.

SummaryUnderstanding hibernation in brown bears (Ursus arctos) can provide insight into many human diseases. During hibernation, brown bears experience states of insulin resistance, physical inactivity, extreme bradycardia, obesity, and the absence of urine production. These states closely mimic human diseases such as type 2 diabetes, muscle atrophy, renal and heart failure, cachexia, and obesity. The reversibility of these states from hibernation to active season allows for the identification of novel mediators with possible therapeutic value for humans. Recent studies have identified genes and pathways that are differentially expressed between active and hibernation seasons. However, little is known about the role of differential expression of gene isoforms on hibernation physiology. To identify both distinct and novel mRNA isoforms, we performed full-length RNA-sequencing (Iso-Seq) on three tissue types from three individuals sampled during both active and hibernation seasons. We combined the long-read data with the reference annotation for an improved transcriptome and mapped RNA-seq data from six individuals to the improved transcriptome to quantify differential isoform usage between tissues and seasons. We identified differentially expressed isoforms in all study tissues and showed that adipose has a high level of differential isoform usage with isoform switching, regardless of whether the genes were differentially expressed. Our analyses provide a comprehensive evaluation of isoform usage between active and hibernation states, revealing that differential isoform usage, even in the absence of differential gene expression, is an important mechanism for modulating genes during hibernation. These findings demonstrate the value of isoform expression studies and will serve as the basis for deeper exploration into hibernation biology.

Obesity is among the most prevalent of health conditions in humans leading to a multitude of metabolic pathologies such as type 2 diabetes and hyperglycemia. However, there are many wild animals that have large seasonal cycles of fat accumulation and loss that do not result in the health consequences observed in obese humans. One example is the grizzly bear (Ursus arctos horribilis) that can have body fat content 40% that is then used as the energy source for hibernation. Previous in vitro studies found that hibernation season adipocytes exhibit insulin resistance and increased lipolysis. Yet, other aspects of cellular metabolism were not addressed, leaving this in vitro model incomplete. Thus, the current studies were performed to determine if the cellular energetic phenotype-measured via metabolic flux-of hibernating bears was retained in cultured adipocytes and to what extent that was due to serum or intrinsic cellular factors. Extracellular acidification rate and oxygen consumption rate were used to calculate proton efflux rate and total ATP defined as both ATP from glycolysis and from mitochondrial respiration. Hibernation adipocytes treated with hibernation serum produced less ATP and exhibited lower maximal respiration and glycolysis rates than active season adipocytes. These effects were reversed with serum from the opposite season. Insulin had little influence on total ATP production and lipolysis in both hibernation and active serum-treated adipocytes. Together, these results suggest that the metabolic suppression occurring in hibernation adipocytes are downstream of insulin signaling and likely due to a combined reduction in mitochondria number and/or function and glycolytic processes. Future elucidation of the serum components and the cellular mechanisms that enable alterations in mitochondrial function could provide a novel avenue for the development of treatments for human metabolic diseases.

Studies of predator feeding ecology commonly focus on energy intake. However, captive predators have been documented to selectively feed to optimize macronutrient intake. As many apex predators experience environmental changes that affect prey availability, limitations on selective feeding can affect energetics and health. We estimated the protein:fat ratio of diets consumed by wild polar bears using a novel isotope-based approach, measured protein:fat ratios selected by zoo polar bears offered dietary choice and examined potential energetic and health consequences of overconsuming protein. Dietary protein levels selected by wild and zoo polar bears were low and similar to selection observed in omnivorous brown bears, which reduced energy intake requirements by 70% compared with lean meat diets. Higher-protein diets fed to zoo polar bears during normal care were concurrent with high rates of mortality from kidney disease and liver cancer. Our results suggest that polar bears have low protein requirements and that limitations on selective consumption of marine mammal blubber consequent to climate change could meaningfully increase their energetic costs. Although bear protein requirements appear lower than those of other carnivores, the energetic and health consequences of protein overconsumption identified in this study have the potential to affect a wide range of taxa.

Animal movements are major determinants of energy expenditure and ultimately the cost–benefit of landscape use. Thus, we sought to understand those costs and how grizzly bears (Ursus arctos) move in mountainous landscapes. We trained captive grizzly bears to walk on a horizontal treadmill and up and down 10% and 20% slopes. The cost of moving upslope increased linearly with speed and slope angle, and this was more costly than moving horizontally. The cost of downslope travel at slower speeds was greater than the cost of traveling horizontally but appeared to decrease at higher speeds. The most efficient walking speed that minimized cost per unit distance was 1.19±0.11 m s−1. However, grizzly bears fitted with GPS collars in the Greater Yellowstone Ecosystem moved at an average velocity of 0.61±0.28 m s−1 and preferred to travel on near-horizontal slopes at twice their occurrence. When traveling uphill or downhill, grizzly bears chose paths across all slopes that were ∼54% less steep and costly than the maximum available slope. The net costs (J kg−1 m−1) of moving horizontally and uphill were the same for grizzly bears, humans and digitigrade carnivores, but those costs were 46% higher than movement costs for ungulates. These movement costs and characteristics of landscape use determined using captive and wild grizzly bears were used to understand the strategies that grizzly bears use for preying on large ungulates and the similarities in travel between people and grizzly bears that might affect the risk of encountering each other on shared landscapes.

Hibernation is characterized by depression of many physiological processes. To determine if this state is reversible in a non-food caching species, we fed hibernating grizzly bears (Ursus arctos horribilis) dextrose for 10 days to replace 53% or 100% of the estimated minimum daily energetic cost of hibernation. Feeding caused serum concentrations of glycerol and ketones (β-hydroxybutyrate) to return to active season levels irrespective of the amount of glucose fed. By contrast, free fatty acids (FFAs) and indices of metabolic rate, such as general activity, heart rate, strength of heart rate circadian rhythm, and insulin sensitivity were restored to approximately 50% of active season levels. Body temperature was unaffected by feeding. To determine the contribution of adipose to the metabolic effects observed after glucose feeding, we cultured bear adipocytes collected at the beginning and end of the feeding and performed metabolic flux analysis. We found a ∼33% increase in energy metabolism after feeding. Moreover, basal metabolism before feeding was 40% lower in hibernation cells compared with fed cells or active cells cultured at 37°C, thereby confirming the temperature independence of metabolic rate. The partial depression of circulating FFAs with feeding likely explains the incomplete restoration of insulin sensitivity and other metabolic parameters in hibernating bears. Further depression of metabolic function is likely to be an active process. Together, the results provide a highly controlled model to examine the relationship between nutrient availability and metabolism on the hibernation phenotype in bears.

Hibernation is characterized by suppression of many physiological processes. To determine if this state is reversible in a non-food caching species, we fed hibernating grizzly bears (Ursus arctos horribilis) glucose for 10 days to replace 53% or 100% of the estimated minimum daily energetic cost of hibernation. Feeding caused serum concentrations of glycerol and ketones (ß-hydroxybutyrate) to return to active season levels irrespective of the amount of glucose fed. By contrast, free-fatty acids and indices of metabolic rate, such as general activity, heart rate, and strength of the daily heart rate rhythm and insulin sensitivity were restored to roughly 50% of active season levels. Body temperature was unaffected by feeding. To determine the contribution of adipose to these metabolic effects of glucose feeding we cultured bear adipocytes collected at the beginning and end of the feeding and performed metabolic flux analysis. We found a roughly 33% increase in energy metabolism after feeding. Moreover, basal metabolism before feeding was 40% lower in hibernation cells compared to fed cells or active cells cultured at 37°C, thereby confirming the temperature independence of metabolic rate. The partial suppression of circulating FFA with feeding likely explains the incomplete restoration of insulin sensitivity and other metabolic parameters in hibernating bears. Further suppression of metabolic function is likely an active process. Together, the results provide a highly controlled model to examine the relationship between nutrient availability and metabolism on the hibernation phenotype in bears.

Revealing the mechanisms underlying the reversible physiology of hibernation could have applications to both human and animal health as hibernation is often associated with disease-like states. The present study uses RNA-sequencing to reveal the tissue and seasonal transcriptional changes occurring in grizzly bears (

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Food sources for North America’s grizzly bear (Ursus arctos horribilis Ord, 1815) population have changed as habitats have fragmented, altering available resources and putting bears in contact with unnatural foods. Bears have evolved mechanisms to tolerate obesity, and do not develop adverse health consequences despite storing massive amounts of body fat. Captive adult grizzly bears were used to determine the effects of dietary fat on health. Group 1 was fed a diet high in polyunsaturated fatty acids (PUFA) wherein 9.5% of available calories came from saturated fatty acids (SFA). Group 2 was fed a diet wherein 28.8% of calories came from SFA. Plasma fatty acids, serum lipid profiles, insulin, inflammatory markers, systolic and diastolic blood pressure, and cardiac function parameters were measured. Serum lipids, SFA, and insulin did not differ between the two groups, although omega-3 fatty acids differed. Bears eating the SFA diet had significantly higher circulating adiponectin, interleukin-7 and interleukin-15, and tumor necrosis factor-alpha. Mild, asymptomatic systolic and diastolic dysfunctions were detected by strain echocardiography in the SFA group. The SFA diet group exhibited higher diastolic arterial pressures. Even though mild metabolic derangements were observed, grizzly bears were remarkably resistant to metabolic effects of diets high in SFA.

Recent studies suggest that bears have relatively high cognitive capabilities. However, cognitive processes and problem-solving abilities remain relatively unexplored in bear species. We studied the capacity for 8 captive brown bears ( Ursus arctos ) to move and use inanimate objects to obtain a food reward. We recorded their behaviors during the problem-solving process using a behavioral ethogram. Three items, a large log, a small log, and a box, were placed in an outdoor enclosure. As the bears progressed through 3 stages of trials, they would need to manipulate the objects and displace them into the proper location and orientation to climb atop to reach a suspended food reward. Completion of the third and final stage was deemed to be evidence of tool use. Six of the 8 bears were capable of tool use. Most bears (>90% of trials) were successful in completing the final stage in

Tri-axial accelerometers have been used to remotely identify the behaviors of a wide range of taxa. Assigning behaviors to accelerometer data often involves the use of captive animals or surrogate species, as their accelerometer signatures are generally assumed to be similar to those of their wild counterparts. However, this has rarely been tested. Validated accelerometer data are needed for polar bears Ursus maritimus to understand how habitat conditions may influence behavior and energy demands. We used accelerometer and water conductivity data to remotely distinguish 10 polar bear behaviors. We calibrated accelerometer and conductivity data collected from collars with behaviors observed from video-recorded captive polar bears and brown bears U. arctos, and with video from camera collars deployed on free-ranging polar bears on sea ice and on land. We used random forest models to predict behaviors and found strong ability to discriminate the most common wild polar bear behaviors using a combination of accelerometer and conductivity sensor data from captive or wild polar bears. In contrast, models using data from captive brown bears failed to reliably distinguish most active behaviors in wild polar bears. Our ability to discriminate behavior was greatest when species- and habitat-specific data from wild individuals were used to train models. Data from captive individuals may be suitable for calibrating accelerometers, but may provide reduced ability to discriminate some behaviors. The accelerometer calibrations developed here provide a method to quantify polar bear behaviors to evaluate the impacts of declines in Arctic sea ice.

The purpose of this study was to investigate changes in expression of known cellular regulators of metabolism during hyperphagia (Sept) and hibernation (Jan) in skeletal muscle and adipose tissue of brown bears and determine whether signaling molecules and transcription factors known to respond to changes in cellular energy state are involved in the regulation of these metabolic adaptations. During hibernation, serum levels of cortisol, glycerol, and triglycerides were elevated, and protein expression and activation of AMPK in skeletal muscle and adipose tissue were reduced. mRNA expression of the co-activator PGC-1α was reduced in all tissues in hibernation whereas mRNA expression of the transcription factor PPAR-α was reduced in the vastus lateralis muscle and adipose tissue only. During hibernation, gene expression of ATGL and CD36 was not altered; however, HSL gene expression was reduced in adipose tissue. During hibernation gene expression of the lipogenic enzyme DGAT in all tissues and the expression of the FA oxidative enzyme LCAD in the vastus lateralis muscle were reduced. Gene and protein expression of the glucose transporter GLUT4 was decreased in adipose tissue in hibernation. Our data suggest that high cortisol levels are a key adaptation during hibernation and link cortisol to a reduced activation of the AMPK/PGC-1α/PPAR-α axis in the regulation of metabolism in skeletal muscle and adipose tissue. Moreover, our results indicate that during this phase of hibernation at a time when metabolic rate is significantly reduced metabolic adaptations in peripheral tissues seek to limit the detrimental effects of unduly large energy dissipation.

There has been considerable emphasis on understanding isotopic discrimination for diet estimation in omnivores. However, discrimination may differ for carnivores, particularly species that consume lipid-rich diets. Here, we examined the potential implications of several factors when using stable isotopes to estimate the diets of bears, which can consume lipid-rich diets and, alternatively, fast for weeks to months. We conducted feeding trials with captive brown bears (Ursus arctos) and polar bears (Ursus maritimus). As dietary lipid content increased to ∼90%, we observed increasing differences between blood plasma and diets that had not been lipid extracted (∆(13)Ctissue-bulk diet) and slightly decreasing differences between plasma δ(13)C and lipid-extracted diet. Plasma Δ(15)Ntissue-bulk diet increased with increasing protein content for the four polar bears in this study and data for other mammals from previous studies that were fed purely carnivorous diets. Four adult and four yearling brown bears that fasted 120 d had plasma δ(15)N values that changed by±2‰. Fasting bears exhibited no trend in plasma δ(13)C. Isotopic incorporation in red blood cells and whole blood was ≥6 mo in subadult and adult bears, which is considerably longer than previously measured in younger and smaller black bears (Ursus americanus). Our results suggest that short-term fasting in carnivores has minimal effects on δ(13)C and δ(15)N discrimination between predators and their prey but that dietary lipid content is an important factor directly affecting δ(13)C discrimination and indirectly affecting δ(15)N discrimination via the inverse relationship with dietary protein content.

Training nonhuman animals in captivity for participation in routine husbandry procedures is believed to produce a lower stress environment compared with undergoing a general anesthetic event for the same procedure. This hypothesis rests largely on anecdotal evidence that the captive subjects appear more relaxed with the trained event. Blood markers of physiological stress responses were evaluated in 4 captive grizzly bears (Ursus arctos horribilis) who were clicker-trained for blood collection versus 4 bears who were chemically immobilized for blood collection. Serum cortisol and immunoglobulin A (IgA) and plasma β-endorphin were measured as indicators of responses to stress. Plasma β-endorphin was not different between the groups. Serum IgA was undetectable in all bears. Serum cortisol was undetectable in all trained bears, whereas chemically immobilized bears had marked cortisol elevations (p .05). The highest cortisol elevations were found in 2 bears with extensive recent immobilization experience. These findings support the use of positive reinforcement training for routine health procedures to minimize anxiety.

The overall health and persistence of whitebark pine is of international concern. Extensive tree mortality and loss of vigor due to the non-native pathogen white pine blister rust (Cronartium ribicola A. Dietr.), mountain pine beetle (Dendroctonus ponderosae Hopkins), altered fire regimes, and climate change endanger the existence of whitebark pine, which is an important food source (pine nuts) for several wildlife species. Prior stable isotope analysis revealed variability in intrinsic water-use efficiency and nutritional status; however, small sample sizes prohibited the identification of geo-climatic effects on those isotopes. We summarize carbon isotope discrimination (Δ13C, a measure of intrinsic water-use efficiency) and the natural abundance of δ13C, δ15N and δ34S in whitebark pine nuts in the Northern Rockies. Spatial differences in geography, soil parent material and climate (e.g., summer mean temperature, number of degree-days below 0 °C, mean annual precipitation, Hargreaves climatic moisture deficit, and the frost-free period) contributed to the low to moderate spatial resolution in selected models (R2 values ranged from 0.24 to 0.51). The importance of soil parent material in all models demonstrated whitebark pine exhibits previously unknown edaphic variation. Identification of the mechanistic drivers contributing to spatial heterogeneity in Δ13C provides an opportunity to select seed sources better suited to optimize long-term survival, vigor and cone production. Matching genetic resources to sites projected to support whitebark pine in future climates would ensure species persistence, while safeguarding an important wildlife food. Suitable wildlife habitat for projected warmer, drier climates was characterized as low values of Δ13C (

Activity sensors are often included in wildlife transmitters and can provide information on the behavior and activity patterns of animals remotely. However, interpreting activity-sensor data relative to animal behavior can be difficult if animals cannot be continuously observed. In this study, we examined the performance of a mercury tip-switch and a tri-axial accelerometer housed in collars to determine whether sensor data can be accurately classified as resting and active behaviors and whether data are comparable for the 2 sensor types. Five captive bears (3 polar [Ursus maritimus] and 2 brown [U. arctos horribilis]) were fitted with a collar specially designed to internally house the sensors. The bears’ behaviors were recorded, classified, and then compared with sensor readings. A separate tri-axial accelerometer that sampled continuously at a higher frequency and provided raw acceleration values from 3 axes was also mounted on the collar to compare with the lower resolution sensors. Both acce...

There is growing interest in the ecological significance of phenological diversity, particularly in how spatially variable resource phenologies (i.e. resource waves) prolong foraging opportunities for mobile consumers. While there is accumulating evidence of consumers moving across landscapes to surf resource waves, there is little data quantifying how phenological tracking influences resource consumption due to the challenge of documenting all the components of this ecological phenomenon (i.e., phenological variation, consumer movement, resource consumption, and consumer fitness). We examined the space use of GPS collared female brown bears to quantify the exploitation of a salmon resource wave by individual bears. We then estimated salmon consumption levels in the same individuals using stable isotope and mercury analyses of hair. We found strong positive relationships between time spent on salmon streams and percent salmon in assimilated diets (R2 = 0.70) and salmon mass consumed (R2 = 0.49). Salmon abundance varied 2.5-fold between study years, yet accounting for salmon abundance did not improve salmon consumption models. Resource abundance generally is viewed as the key variable controlling consumption levels and food web dynamics. However, our results suggest that in intact watersheds of coastal Alaska with abundant salmon runs, interannual variation in salmon abundance likely has less effect on salmon consumption than individual variation in bear foraging behavior. The results complement previous work to demonstrate the importance of phenological variation on bear foraging behavior and fitness.

Ursids are the largest mammals to retain a plantigrade posture. This primitive posture has been proposed to result in reduced locomotor speed and economy relative to digitigrade and unguligrade species, particularly at high speeds. Previous energetics research on polar bears (Ursus maritimus) found locomotor costs were more than double predictions for similarly sized quadrupedal mammals, which could be a result of their plantigrade posture or due to adaptations to their Arctic marine existence. To evaluate whether polar bears are representative of terrestrial ursids or distinctly uneconomical walkers, this study measured the mass-specific metabolism, overall dynamic body acceleration, and gait kinematics of polar bears and grizzly bears (Ursus arctos) trained to rest and walk on a treadmill. At routine walking speeds, we found polar bears and grizzly bears exhibited similar costs of locomotion and gait kinematics, but differing measures of overall dynamic body acceleration. Minimum cost of transport while walking in the two species (2.21 J kg−1 m−1) was comparable to predictions for similarly sized quadrupedal mammals, but these costs doubled (4.42 J kg−1 m−1) at speeds ≥5.4 km h−1. Similar to humans, another large plantigrade mammal, bears appear to exhibit a greater economy while moving at slow speeds.

Increased land use by polar bears (Ursus maritimus) due to climate‐change‐induced reduction of their sea‐ice habitat illustrates the impact of climate change on species distributions and the difficulty of conserving a large, highly specialized carnivore in the face of this global threat. Some authors have suggested that terrestrial food consumption by polar bears will help them withstand sea‐ice loss as they are forced to spend increasing amounts of time on land. Here, we evaluate the nutritional needs of polar bears as well as the physiological and environmental constraints that shape their use of terrestrial ecosystems. Only small numbers of polar bears have been documented consuming terrestrial foods even in modest quantities. Over much of the polar bear's range, limited terrestrial food availability supports only low densities of much smaller, resident brown bears (Ursus arctos), which use low‐quality resources more efficiently and may compete with polar bears in these areas. Where consumption of terrestrial foods has been documented, polar bear body condition and survival rates have declined even as land use has increased. Thus far, observed consumption of terrestrial food by polar bears has been insufficient to offset lost ice‐based hunting opportunities but can have ecological consequences for other species. Warming‐induced loss of sea ice remains the primary threat faced by polar bears.

Mammalian hibernation has intrigued scientists due to extreme variations in normal seasonal physiological homeostasis. Numerous species manifest a hibernation phenotype although the characteristics of the hypometabolic state can be quite different. Ground squirrels (e.g., Sciuridae) are often considered the prototypical hibernator as individuals in this genus transition from an active, euthermic state (37 °C) to a nonresponsive hibernating state where torpid body temperature commonly falls to 3-5 °C. However, the hibernating state is not continuous as periodic warming and arousals occur. In contrast, the larger hibernators of genus Ursus are less hypothermic (body temperatures decline from approximately 37°-33 °C), are more reactive, and cyclical arousals do not occur. Both species dramatically reduce cardiac output during hibernation from the active state (bears ~75 % reduction in bears and ~97 % reduction in ground squirrels), and both species demonstrate hypokinetic atrial chamber activity. However, there are several important differences in cardiac function between the two groups during hibernation. Left ventricular diastolic filling volumes and stroke volumes do not differ in bears between seasons, but increased diastolic and stroke volumes during hibernation are important contributors to cardiac output in ground squirrels. Decreased cardiac muscle mass and increased ventricular stiffness have been found in bears, whereas ground squirrels have increased cardiac muscle mass and decreased ventricular stiffness during hibernation. Molecular pathways of cardiac muscle plasticity reveal differences between the species in the modification of sarcomeric proteins such as titin and α myosin heavy chain during hibernation. The differences in hibernation character are likely to account for the alternative cardiac phenotypes and functional strategies manifested by the two species. Molecular investigation coupled with better knowledge of seasonal physiological alterations is dramatically advancing our understanding of small and large hibernators and their evolutionary differences.

We quantified the amount, spatial distribution, and importance of salmon (Oncorhynchus spp.)-derived nitrogen (N) by brown bears (Ursus arctos) on the Kenai Peninsula, Alaska. We tested and confirmed the hypothesis that the stable isotope signature (δ15N) of N in foliage of white spruce (Picea glauca) was inversely proportional to the distance from salmon-spawning streams (r=–0.99 and P

We used enzyme immunoassays to measure testosterone, progesterone, estradiol and cortisol concentrations in serial hair samples collected from captive adult brown bears. Reproductive hormone levels changed throughout the year in step with different reproductive events underscoring the strong potential for these measurement to be used to augment non-invasive genetic sampling., 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.

The confluence of obesity and diabetes as a worldwide epidemic necessitates the discovery of new therapies. Success in this endeavor requires translatable preclinical studies, which traditionally employ rodent models. As an alternative approach, we explored hibernation where obesity is a natural adaptation to survive months of fasting. Here we report that grizzly bears exhibit seasonal tripartite insulin responsiveness such that obese animals augment insulin sensitivity but only weeks later enter hibernation-specific insulin resistance (IR) and subsequently reinitiate responsiveness upon awakening. Preparation for hibernation is characterized by adiposity coupled to increased insulin sensitivity via modified PTEN/AKT signaling specifically in adipose tissue, suggesting a state of "healthy" obesity analogous to humans with PTEN haploinsufficiency. Collectively, we show that bears reversibly cope with homeostatic perturbations considered detrimental to humans and describe a mechanism whereby IR functions not as a late-stage metabolic adaptation to obesity, but rather a gatekeeper of the fed-fasting transition.

Use of naturally occurring stable isotopes to estimate assimilated diet of bears is one of the single greatest breakthroughs in nutritional ecology during the past 20 years. Previous research in the Greater Yellowstone Ecosystem (GYE), USA, established a positive relationship between the stable isotope of sulfur (δ34S) and consumption of whitebark pine (Pinus albicaulis) seeds. That work combined a limited sample of hair, blood clots, and serum. Here we use a much larger sample to reassess those findings. We contrasted δ34S values in spring hair and serum with abundance of seeds of whitebark pine in samples collected from grizzly (Ursus arctos) and American black bears (U. americanus) in the GYE during 2000–2010. Although we found a positive relationship between δ34S values in spring hair and pine seed abundance for grizzly bears, the coefficients of determination were small (R2 ≤ 0.097); we failed to find a similar relationship with black bears. Values of δ34S in spring hair were larger in black bears and δ34S values in serum of grizzly bears were lowest in September and October, a time when we expect δ34S to peak if whitebark pine seeds were the sole source of high δ34S. The relationship between δ34S in bear tissue and the consumption of whitebark pine seeds, as originally reported, may not be as clean a method as proposed. Data we present here suggest other foods have high values of δ34S, and there is spatial heterogeneity affecting the δ34S values in whitebark pine, which must be addressed. Published 2014. This article is a U.S. Government work and is in the public domain in the USA.

Many animals consume mixed diets that maximize their fitness by optimizing macronutrient intake. We tested whether brown bears (Ursus arctos), generalist omnivores that hibernate, regulated their diet to a common nutrient target, achieved a nutrient target related to fitness, and selected a nutrient target that differed between seasons and from other species with differing life histories. When given unlimited access to 2 or 3 highly digestible foods containing primarily protein, carbohydrate, or lipid, brown bears selected mixed diets in which protein provided 17% ± 4% SD of the metabolizable energy and 22% ± 6% of the dry matter. This dietary protein content maximized the rate of gain per unit of energy consumed, is similar to the level preferred by other omnivores, and is less than that preferred by obligate carnivores. Between seasons, bears selected similar dietary protein levels, although the proportion of lipid was higher during the fall than during the spring. Bears strongly preferred lipids over carbohydrates, as did other carnivores, but they used lipids and carbohydrates with equal efficiency to produce a dietary protein content that maximized mass gain per unit of energy intake. Thus, dietary sources of lipids and carbohydrates play an interchangeable and important role in determining the productivity of bears that goes beyond their role in providing energy.

Spawning cutthroat trout (Oncorhynchus clarkii bouvieri) were historically abundant within tributary streams of Yellowstone Lake within Yellowstone National Park and were a highly digestible source of energy and protein for Yellowstone's grizzly bears (Ursus arctos) and black bears (U. americanus). The cutthroat trout population has subsequently declined since the introduction of non-native lake trout (Salvelinus namaycush), and in response to effects of drought and whirling disease (Myxobolus cerebralis). The trout population, duration of spawning runs, and indices of bear use of spawning streams had declined in some regions of the lake by 1997–2000. We initiated a 3-year study in 2007 to assess whether numbers of spawning fish, black bears, and grizzly bears within and alongside stream corridors had changed since 1997–2000. We estimated numbers of grizzly bears and black bears by first compiling encounter histories of individual bears visiting 48 hair-snag sites along 35 historically fished streams. We analyzed DNA encounter histories with Pradel-recruitment and Jolly-Seber (POPAN) capture-mark-recapture models. When compared to 1997–2000, the current number of spawning cutthroat trout per stream and the number of streams with cutthroat trout has decreased. We estimated that 48 (95% CI = 42–56) male and 23 (95% CI = 21–27) female grizzly bears visited the historically fished tributary streams during our study. In any 1-year, 46 to 59 independent grizzly bears (8–10% of estimated Greater Yellowstone Ecosystem population) visited these streams. When compared with estimates from the 1997 to 2000 study and adjusted for equal effort, the number of grizzly bears using the stream corridors decreased by 63%. Additionally, the number of black bears decreased between 64% and 84%. We also document an increased proportion of bears of both species visiting front-country (i.e., near human development) streams. With the recovery of cutthroat trout, we suggest bears that still reside within the Lake basin will readily use this high-quality food resource. © 2014 The Wildlife Society.

a b s t r a c t Brown bears give birth and nurture their young for the first 3-5 months while fasting in hibernation. During this period, bears use body reserves to support the energy and protein costs of reproduction. Limited reserves may restrict reproductive investment. We developed a model to assess the energetic costs of reproduction during hibernation and to determine how maternal condition, length of lactation, litter size, and length of hibernation affect brown bear reproductive success. Increasing litter size by one cub was more costly than increasing the lactation period by 14 days. For hibernating females, ∼73% of their mass loss during lactation was lean mass. Minimum levels of fat reserves necessary to support reproduction varied from 19% to 33% depending on number of cubs and length of lactation while denned. Reproduction was not possible if body fat content was below 19% and length of hibernation was over 120 days. While the importance of fat or energy reserves at the beginning of hibernation has been long recognized, the importance of consuming high-protein foods after hibernation may be equally important. Therefore, more attention should be given to understanding pre- and post-hibernation nutritional factors affecting reproduction and the trade-offs between available food resources, maternal condition, and reproductive investment and success in wild bear populations. © 2013 Published by Elsevier B.V.

The Greater Yellowstone Ecosystem (GYE) has experienced changes in the distribution and availability of grizzly bear (Ursus arctos) food resources in recent decades. The decline of ungulates, fish, and whitebark pine seeds (Pinus albicaulis) has prompted questions regarding their ability to adapt. We examined body composition and diet of grizzly bears using bioelectrical impedance and stable isotopes to determine if 1) we can detect a change in diet quality associated with the decline in either ungulates or whitebark pine, and 2) the combined decline in ungulates, fish, and pine seeds resulted in a change in grizzly bear carrying capacity in the GYE. We contrasted body fat and mass in grizzly bears with a potential competitor, the American black bear (Ursus americanus), to address these questions. Grizzly bears assimilated more meat into their diet and were in better body condition than black bears throughout the study period, indicating the decline in ungulate resources did not affect grizzly bears more than black bears. We also found no difference in autumn fat levels in grizzly bears in years of good or poor pine seed production, and stable isotope analyses revealed this was primarily a function of switching to meat resources during poor seed-producing years. This dietary plasticity was consistent over the course of our study. We did not detect an overall downward trend in either body mass or the fraction of meat assimilated into the diet by grizzly bears over the past decade, but we did detect a downward trend in percent body fat in adult female grizzly bears after 2006. Whether this decline is an artifact of small sample size or due to the population reaching the ecological carrying capacity of the Yellowstone ecosystem warrants further investigation. © 2013 The Wildlife Society