Jon M. Arnemo

The genomic signature of dog domestication reveals adaptation to a starch-rich diet

The domestication of dogs was an important episode in the development of human civilization. The precise timing and location of this event is debated and little is known about the genetic changes that accompanied the transformation of ancient wolves into domestic dogs. Here we conduct whole-genome resequencing of dogs and wolves to identify 3.8 million genetic variants used to identify 36 genomic regions that probably represent targets for selection during dog domestication. Nineteen of these regions contain genes important in brain function, eight of which belong to nervous system development pathways and potentially underlie behavioural changes central to dog domestication. Ten genes with key roles in starch digestion and fat metabolism also show signals of selection. We identify candidate mutations in key genes and provide functional support for an increased starch digestion in dogs relative to wolves. Our results indicate that novel adaptations allowing the early ancestors of modern dogs to thrive on a diet rich in starch, relative to the carnivorous diet of wolves, constituted a crucial step in the early domestication of dogs.

Risk of capture-related mortality in large free-ranging mammals: experiences from Scandinavia

Abstract Chemical capture and anaesthesia of free-ranging mammals will always involve some risk of mortality even in healthy animals. Deaths may be directly or indirectly attributable to the anaesthetic event itself (e.g. drug overdose, drowning during induction and dart trauma) or may be caused by secondary effects from the capture (e.g. stress, myopathy, trauma or instrumentation with radio-transmitters). In long-term research projects on five major wildlife species in Scandinavia, the capture-related mortality rates (number of captures) were: moose Alces alces 0.7% (N = 2,816), brown bears Ursus arctos 0.9% (N = 1,079), wolverines Gulo gulo 2.8% (N = 461), Eurasian lynx Lynx lynx 3.9% (N = 380), and gray wolves Canis lupus 3.4% (N = 89). We suggest that wildlife professionals should strive for a zero mortality rate but adopt the standard that a mortality rate of > 2% probably should not be accepted in any large mammalian species. This can be achieved by: 1) using an experienced professional capture team, 2) developing and following a capture protocol specific to each species, and 3) requiring that a mortality assessment be undertaken after any capture-related death. This assessment should re-evaluate the capture protocol, including how changes in anaesthetics and methodological approaches could have prevented the mortality.

The Gut Microbiota Modulates Energy Metabolism in the Hibernating Brown Bear Ursus arctos

Hibernation is an adaptation that helps many animals to conserve energy during food shortage in winter. Brown bears double their fat depots during summer and use these stored lipids during hibernation. Although bears seasonally become obese, they remain metabolically healthy. We analyzed the microbiota of free-ranging brown bears during their active phase and hibernation. Compared to the active phase, hibernation microbiota had reduced diversity, reduced levels of Firmicutes and Actinobacteria, and increased levels of Bacteroidetes. Several metabolites involved in lipid metabolism, including triglycerides, cholesterol, and bile acids, were also affected by hibernation. Transplantation of the bear microbiota from summer and winter to germ-free mice transferred some of the seasonal metabolic features and demonstrated that the summer microbiota promoted adiposity without impairing glucose tolerance, suggesting that seasonal variation in the microbiota may contribute to host energy metabolism in the hibernating brown bear.

Physiologic Evaluation of Capture and Anesthesia with Medetomidine–Zolazepam–Tiletamine in Brown Bears (Ursus arctos)

Abstract Physiologic variables during anesthesia with medetomidine–zolazepam–tiletamine were evaluated in 52 free-ranging brown bears (Ursus arctos) darted from a helicopter and in six captive brown bears darted at a zoo. During anesthesia, rectal temperature, respiratory rate, heart rate, and pulse oximetry derived hemoglobin oxygen saturation were recorded. Arterial blood samples were collected and immediately analyzed for evaluation of pulmonary gas exchange, acid–base status, and selected hematologic and plasma variables. At the end of anesthesia, atipamezole was administered intramuscularly at five times the medetomidine dose. Capture-induced hyperthermia and lactic acidemia were documented in free-ranging bears. Hypoxemia during anesthesia was documented in both free-ranging and captive bears. In free-ranging bears, rectal temperature, heart rate, lactate, hematocrit, and hemoglobin decreased significantly during anesthesia, whereas partial pressure of arterial carbon dioxide, pH, potassium, and glucose increased. Yearlings had a significantly higher heart rate, pH, base excess, bicarbonate, and glucose, and had a significantly lower rectal temperature, sodium, hematocrit, and hemoglobin when compared with subadult and adult brown bears. In conclusion, alterations in pulmonary gas exchange and acid–base status in brown bears during anesthesia with medetomidine–zolazepam–tiletamine with the doses and capture methods used in this study were identified. Oxygen supplementation is recommended to counteract hypoxemia during anesthesia.

Capture, anesthesia, and disturbance of free-ranging brown bears (Ursus arctos) during hibernation.

We conducted thirteen immobilizations of previously collared hibernating two- to four-year-old brown bears (Ursus arctos) weighing 21–66 kg in central Sweden in winter 2010 and 2011 for comparative physiology research. Here we report, for the first time, an effective protocol for the capture and anesthesia of free-ranging brown bears during hibernation and an assessment of the disturbance the captures caused. Bears were darted in anthill, soil, or uprooted tree dens on eleven occasions, but two bears in rock dens fled and were darted outside the den. We used medetomidine at 0.02–0.06 mg/kg and zolazepam-tiletamine at 0.9–2.8 mg/kg for anesthesia. In addition, ketamine at 1.5 mg/kg was hand-injected intramuscularly in four bears and in six it was included in the dart at 1.1–3.0 mg/kg. Once anesthetized, bears were removed from the dens. In nine bears, arterial blood samples were analyzed immediately with a portable blood gas analyzer. We corrected hypoxemia in seven bears (PaO2 57–74 mmHg) with supplemental oxygen. We placed the bears back into the dens and antagonized the effect of medetomidine with atipamezole. Capturing bears in the den significantly increased the risk of den abandonment. One of twelve collared bears that were captured remained at the original den until spring, and eleven, left their dens (mean ± standard deviation) 3.2±3.6 (range 0.5–10.5) days after capture. They used 1.9±0.9 intermediate resting sites, during 6.2±7.8 days before entering a new permanent den. The eleven new permanent dens were located 730±589 m from the original dens. We documented that it was feasible and safe to capture hibernating brown bears, although they behaved differently than black bears. When doing so, researchers should use 25% of the doses used for helicopter darting during the active period and should consider increased energetic costs associated with den abandonment.

Adipose-derived stem cells from the brown bear (Ursus arctos) spontaneously undergo chondrogenic and osteogenic differentiation in vitro.

In the den, hibernating brown bears do not develop tissue atrophy or organ damage, despite almost no physical activity. Mesenchymal stem cells could play an important role in tissue repair and regeneration in brown bears. Our objective was to determine if adipose tissue-derived stem cells (ASCs) can be recovered from wild Scandinavian brown bears and characterize their differentiation potential. Following immobilization of wild brown bears 7-10 days after leaving the den in mid-April, adipose tissue biopsies were obtained. ASCs were recovered from 6 bears, and shown to be able to undergo adipogenesis and osteogenesis in monolayer cultures and chondrogenesis in pellet cultures. Remarkably, when grown in standard cell culture medium in monolayer cultures, ASCs from yearlings spontaneously formed bone-like nodules surrounded by cartilaginous deposits, suggesting differentiation into osteogenic and chondrogenic lineages. This ability appears to be lost gradually with age. This is the first study to demonstrate stem cell recovery and growth from brown bears, and it is the first report of ASCs spontaneously forming extracellular matrix characteristic of bone and cartilage in the absence of specific inducers. These findings could have implications for the use of hibernating brown bears as a model to study disuse osteoporosis.

SEROSURVEY FOR CANINE DISTEMPER VIRUS, CANINE ADENOVIRUS, LEPTOSPIRA INTERROGANS, AND TOXOPLASMA GONDII IN FREE-RANGING CANIDS IN SCANDINAVIA AND SVALBARD

Prevalence of antibodies reactive to canine distemper virus (CDV), canine adenovirus type 1 (CAV-1), Leptospira interrogans serovars Canicola and Icterohaemorrhagiae, and Toxoplasma gondii were examined in free-ranging Scandinavian canids. Sampling included 275 red foxes (Vulpes vulpes) from mainland Norway, 60 arctic foxes (Vulpes lagopus) from the high-arctic islands of Svalbard, and 98 wolves (Canis lupus) from the joint Swedish-Norwegian population. Methods used included virus neutralization tests for CDV and CAV-1, a microscopic agglutination test for L. interrogans, and a direct agglutination test for T. gondii. High prevalence of antibody to CAV-1 was identified in red foxes (59.6%), wolves (67.7%), and arctic foxes (37.8%). The prevalence of antibody to CDV varied between 9.6% and 12.3% in the three species. Antibodies to L. interrogans serovar Icterohaemorrhagiae were found in 9.9% of the red foxes and 8.4% of the wolves sampled, whereas no antibody-positive arctic foxes were found. All animals were antibody-negative for L. interrogans serovar Canicola. Antibodies to T. gondii were found in 66.9, 51.7, and 18.6% of red foxes, arctic foxes and wolves, respectively. Significantly more adults than juveniles were antibody-positive for CDV in red foxes and arctic foxes, for CAV-1 in wolves, and for T. gondii in red foxes and wolves. There was a general tendency for adult female red foxes to have a higher prevalence of antibodies for CDV than adult males; this difference was statistically significant. The results indicate that CDV and CAV-1 are endemic in red foxes and wolves on the Scandinavian mainland and in arctic foxes on Svalbard. Although infection with L. interrogans serovar Icterohaemorrhagiae was relatively common in wild canids on mainland Norway, it was not found on Svalbard, where the maintenance host (Rattus norvegicus) is absent. All three species are commonly exposed to T. gondii through predation on infected intermediate hosts.

PLASMA BIOCHEMICAL VALUES FROM APPARENTLY HEALTHY FREE-RANGING POLAR BEARS FROM SVALBARD

To establish reference values for free-ranging polar bears (Ursus maritimus) at Svalbard, Norway, plasma samples from 15 females and 20 males were analyzed for 28 blood biochemistry parameters. Animals were chemically immobilized (Zoletil®: tiletamine and zolazepam) on land at Barentsøya, Edgeøya, and the eastern coast of Spitsbergen in August 1998. All bears were apparently healthy, with ages ranging from 1–22 yr. Females had almost two times higher levels of lipase than males. Several parameters varied with age. Levels of alkaline phosphatase (ALP) and calcium (Ca) decreased with age, being significantly higher in young individuals (<6 yr) compared to middle-aged (6–13 yr) and older bears (>13 yr). Globulin was lower in animals <6 yr of age than in animals >13 yr of age, while the opposite was the case for albumin. Levels of ALP, Ca, and potassium decreased with age. We found no significant changes in total protein correlated to age, but total protein levels were higher in obese compared to lean individuals. Further, total protein levels were slightly lower and had greater variation compared to data from polar bears in captivity, which may reflect food availability for the latter group. The mean ratio between urea and creatinine was 10.9 and indicated these bears were fasting. These data provide a baseline from which to compare biochemical parameters in captive and free-ranging polar bears and will be especially valuable for future studies of polar bears at Svalbard.

Factors Affecting Date of Implantation, Parturition, and Den Entry Estimated from Activity and Body Temperature in Free-Ranging Brown Bears

Knowledge of factors influencing the timing of reproduction is important for animal conservation and management. Brown bears (Ursus arctos) are able to vary the birth date of their cubs in response to their fat stores, but little information is available about the timing of implantation and parturition in free-ranging brown bears. Body temperature and activity of pregnant brown bears is higher during the gestation period than during the rest of hibernation and drops at parturition. We compared mean daily body temperature and activity levels of pregnant and nonpregnant females during preimplantation, gestation, and lactation. Additionally we tested whether age, litter size, primiparity, environmental conditions, and the start of hibernation influence the timing of parturition. The mean date of implantation was 1 December (SD = 12), the mean date of parturition was 26 January (SD = 12), and the mean duration of the gestation period was 56 days (SD = 2). The body temperature of pregnant females was higher during the gestation and lactation periods than that of nonpregnant bears. The body temperature of pregnant females decreased during the gestation period. Activity recordings were also used to determine the date of parturition. The parturition dates calculated with activity and body temperature data did not differ significantly and were the same in 50% of the females. Older females started hibernation earlier. The start of hibernation was earlier during years with favorable environmental conditions. Dates of parturition were later during years with good environmental conditions which was unexpected. We suggest that free-ranging pregnant brown bears in areas with high levels of human activities at the beginning of the denning period, as in our study area, might prioritize investing energy in early denning than in early parturition during years with favorable environmental conditions, as a strategy to prevent disturbances caused by human.

Brown Bears (Ursus arctos) Seem Resistant to Atherosclerosis ­Despite Highly Elevated Plasma Lipids during Hibernation and Active State

Hibernation is an extreme physiological challenge for the brown bear (Ursus arctos) in which metabolism is based mainly on lipids. The study objective was to compare plasma lipids in hibernating and active free‐ranging brown bears and relate them to arterial histopathology. Blood was drawn from seven immobilized free‐ranging brown bears (three females, 2–3 years old) during hibernation in February and from the same bears while active in June and analyzed by enzymatic and automated hematology methods within 48 hours of sampling. Left anterior descending coronary arteries and aortic arches from 12 bears (six females, 1.5–12 years old) killed in hunting were examined by histopathology. Total plasma cholesterol decreased from hibernation to the active period (11.08 ± 1.04 mmol/L vs. 7.89 ± 1.96 mmol/L, P= 0.0028) as did triglyceride (3.16 ± 0.62 mmol/L vs. 1.44 ± 0.27 mmol/L, P= 0.00012) and LDL cholesterol (4.30 ± 0.71 mmol/L vs. 2.02 ± 1.03 mmol/L, P= 0.0075), whereas HDL cholesterol was unchanged. No atherosclerosis, fatty streaks, foam cell infiltration, or inflammation were seen in any arterial samples. Brown bears tolerate elevated cholesterol levels, obesity, physical inactivity, and circulatory slow flow during hibernation without signs of atherosclerosis. This species might serve as a reverse translational model for atherosclerosis resistance. Clin Trans Sci 2012; Volume 5: 269–272