Tubulopapillary mammary carcinoma in a brown bear (Ursus arctos)

A 28-yr-old, nulliparous female brown bear (Ursus arctos) at the Karacabey Ovakurusu Bear Sanctuary presented with an enlargement of the mammary gland. Three other nodules were also noted in the proximity of the mammary gland and over the vulva. Clinical, hematologic, ultrasonographic, and radiologic examinations were performed; the enlarged mammary gland was removed and the other masses were also excised. Histopathologic examination revealed tubulopapillary carcinoma of the mammary gland, and the other masses were diagnosed as epidermoid cysts. This is the first reported case of tubulopapillary mammary carcinoma accompanied by epidermoid cysts in a bear.     

Fallot's tetralogy in a European brown bear (Ursus arctos)

A congenital heart malformation was diagnosed in a free-ranging adult female European brown bear (Ursus arctos) found dead due to intraspecific violence. At necropsy, the heart had all four features of Fallot's tetralogy. No further malformations were identified in the animal, which was of normal size and weight for its age, in normal body condition, and had probably borne young. The heart condition was considered to have contributed to death during an attack by another bear.     

The mating system of the brown bear Ursus arctos

• 1 Research on mating systems and reproductive strategies is valuable for providing ethological knowledge, important for the management and conservation of a species, and in a broader sense, important for biodiversity conservation.
• 2 We reviewed the literature to document the mating system of the brown bear Ursus arctos. We determined that many aspects of the reproduction of the brown bear remain unclear, including (i) biological aspects, such as hormone and oestrous cycling, sperm competition, mate choice, sexually selected infanticide, etc. and (ii) human impacts on the mating system, occurring when humans alter population size and structure, through, for example, hunting or habitat degradation.
• 3 We considered three mating system classification frameworks from the literature ( Emlen & Oring 1977 , Clutton‐Brock 1989 , Shuster & Wade 2003 ) and applied various brown bear populations to them. We did this (i) to document the plasticity of the mating system of the brown bear, and (ii) to find commonalities among the reported mating system classifications in order to provide a general and common classification of the brown bear's mating system.
• 4 The mating system of the brown bear can, in general, be classed as ‘polygamous’. Subclassifications can nevertheless be valuable on smaller spatial scales.
• 5 Within the polygamous mating system of the brown bear, biological aspects and human impacts can influence reproductive strategies at the individual and population level. Mating system classification frameworks often lack a common terminology, which contributes to the variety of published classifications of the mating system of the brown bear.

     

Seasonal patterns in the physiology of the European brown bear (Ursus arctos arctos) in Finland

The physiological indicators such as body temperature, blood chemistry and hematology of seven European brown bears (Ursus arctos arctos) were used in the present study. They were kept in either the Zoological Garden of University of Oulu (65°N, 25°24'E) or the Ranua Zoological Garden approx. 150 km NE of Oulu. Transmitters with a temperature-dependent pulse rate were implanted subcutaneously or into the abdominal cavity under anesthesia. Our data indicate that the body temperature of the bear decreases during the winter sleep to 4–5°C below the normal level (37.0–37.5°C). The lowest values, 33.1–33.3°C, were measured several times in midwinter. Hematocrit, hemoglobin and erythrocyte counts seem to be higher, and the leucocyte count lower during the denning period than in the awake bear. Plasma N-wastes were lower during the winter sleep than before or after it. The analysed blood parameters showed that plasma catecholamines and thyroid hormones decreased in the fall.     

Non-invasive genetic study of the endangered Cantabrian brown bear (Ursus arctos)

The Brown Bear (Ursus arctos) population present in the Cantabrian Mountains has suffered a dramatic decline in recent centuries and is now threatened with extinction. This situation has led to the development and implementation of a species recovery plan. To accomplish this plan, we need to improve our knowledge about the ecology, demography and genetics of this population. This paper presents the genetic analysis of the Cantabrian brown bear population using non-invasive samples (faeces and hairs) collected between 2004 and 2006. It was necessary to optimize a set of 18 microsatellite loci and a sex marker (several new multiplex reactions were developed) to obtain a suitable probability of identity among genotypes to work with this small, deeply structured population. Genotyping of 48 individuals was carried out using a two-step PCR protocol to increase the quality of the multilocus genotypes. Validation of genotypes was performed using a multi-tube approach combined with different software programmes to measure their error rate and reliability. Diversity in the Cantabrian population was low (H _e = 0.51) and the population was markedly subdivided into two subpopulations (western and eastern) without current gene flow between them. The level of divergence between the two subpopulations (F _st = 0.41) and the extremely low diversity in the eastern group (H _e = 0.25) indicate that this has had an extremely low effective population size and had been isolated from the main group during the last century. Connectivity between the two subpopulations will be of prime importance for the long-term survival of this species in the Cantabrian Mountains.     

Geographic and genetic boundaries of brown bear (Ursus arctos) population in the Caucasus

The taxonomic status of brown bears in the Caucasus remains unclear. Several morphs or subspecies have been identified from the morphological (craniological) data, but the status of each of these subspecies has never been verified by molecular genetic methods. We analysed mitochondrial DNA sequences (control region) to reveal phylogenetic relationships and infer divergence time between brown bear subpopulations in the Caucasus. We estimated migration and gene flow from both mitochondrial DNA and microsatellite allele frequencies, and identified possible barriers to gene flow among the subpopulations. Our suggestion is that all Caucasian bears belong to the nominal subspecies of Ursus arctos. Our results revealed two genetically and geographically distinct maternal haplogroups: one from the Lesser Caucasus and the other one from the Greater Caucasus. The genetic divergence between these haplogroups dates as far back as the beginning of human colonization of the Caucasus. Our analysis of the least‐cost distances between the subpopulations suggests humans as a major barrier to gene flow. The low genetic differentiation inferred from microsatellite allele frequencies indicates that gene flow between the two populations in the Caucasus is maintained through the movements of male brown bears. The Likhi Ridge that connects the Greater and Lesser Caucasus mountains is the most likely corridor for this migration.     

Improved noninvasive genotyping method: application to brown bear (Ursus arctos) faeces

We redesigned new microsatellite primers and one sex‐specific primer for amplification of faecal DNA from brown bears (Ursus arctos). We also combined a semi‐nested polymerase chain reaction (PCR) with a newly developed multiplex preamplification method in order to increase the quality of the amplified DNA fragments. In comparison with a conventional PCR approach, the genotyping error rate was substantially reduced and the amplification rate was increased. This new approach could be transposed to other species where conventional PCR methods experience low success due to limited DNA concentration and/or quality.     

Split parturition observed in a captive North American brown bear (Ursus arctos)

Reproductive physiology in North American ursids is characterized by mating from spring to early summer, delayed implantation, and birth during hibernation. During spring 2008, a captive adult female brown bear was mated with two adult males. Pregnancy was determined by elevated progesterone concentrations during late fall before hibernation. Two male cubs were born on December 31, 2008, and a third female cub was born 17 days later on January 16. All were successfully raised and all were confirmed to have identical paternity. When normalized to age, cub growth rates did not differ. To our knowledge, this is the first documented case of markedly different birth dates in a single litter of brown bear cubs.     

An evaluation of long-term preservation methods for brown bear (Ursus arctos) faecal DNA samples

Relatively few large-scale faecal DNA studieshave been initiated due to difficulties inamplifying low quality and quantity DNAtemplate. To improve brown bear faecal DNA PCRamplification success rates and to determinepost collection sample longevity, fivepreservation methods were evaluated: 90%ethanol, DETs buffer, silica-dried, oven-driedstored at room temperature, and oven-driedstored at –20 °C. Preservationeffectiveness was evaluated for 50 faecalsamples by PCR amplification of a mitochondrialDNA (mtDNA) locus (146 bp) and a nuclear DNA(nDNA) locus (200 bp) at time points of oneweek, one month, three months and six months. Preservation method and storage timesignificantly impacted mtDNA and nDNAamplification success rates. For mtDNA, allpreservation methods had 75% success atone week, but storage time had a significantimpact on the effectiveness of the silicapreservation method. Ethanol preserved sampleshad the highest success rates for both mtDNA(86.5%) and nDNA (84%). Nuclear DNAamplification success rates ranged from 26–88%, and storage time had a significant impacton all methods but ethanol. Preservationmethod and storage time should be importantconsiderations for researchers planningprojects utilizing faecal DNA. We recommendpreservation of faecal samples in 90% ethanolwhen feasible, although when collecting inremote field conditions or for both DNA andhormone assays a dry collection method may beadvantageous.     

Range and local population densities of brown bear Ursus arctos in Slovenia

Solid understanding of species’ range and local population densities is important for successful wildlife management and research. Specific behavioral and ecological characteristics make brown bear Ursus arctos a difficult species to study. We present a map of range and local population densities of brown bears in Slovenia, made with the use of a new approach similar to voting classifications based on a combination of four datasets: Global Positioning System telemetry data, records of bear removals, systematic and opportunistic direct observations and signs of bear presence, and noninvasive genetic samples. Results indicate that the majority of bears in Slovenia live in Dinaric Mountains in the southern part of the country where local bear population densities exceed 40 bears/100 km². This is one of the highest population densities reported so far for this species worldwide. Population densities decrease towards the north (Alpine region) and are very low along the border with Italy and Austria where almost no females are present. This explains slow past and present expansion of this transboundary bear population into the Alps and should be considered in future bear re-colonization management strategies. Results also showed that data from observations and removals overestimate bear population densities at low values, while mortality and genetic data overestimate population densities in areas with more people. Nevertheless, all data types appeared useful for describing the general bear distribution patterns. Similar approach could be applied to studies of other charismatic or game species, for which several types of data are often available.     

On the association of giant short-faced bear (Arctodus simus) and brown bear (Ursus arctos) in late Pleistocene North America

Climate change and human impacts are often implicated in Quaternary megafaunal extinctions. The discovery of associated remains of extinct giant short-faced bears (Arctodus simus) and invading brown bears (Ursus arctos) raises the possibility of competition as another potential factor. We describe fossil remains of both genera from Pellucidar Cave, Vancouver Island, Canada. Analyses of ancient mitochondrial DNA support the identifications of post-cranial brown bear specimens and assign these bears to Clade 4. Our results are consistent with the migration of brown bears from Eastern Beringia to the contiguous United States before the Last Glacial Maximum (LGM) and to Vancouver Island as environmental conditions became favorable after the LGM. Radiocarbon age estimates on these specimens indicate the presence of giant short-faced bears approximately 13.5 thousand calibrated years before present (cal. ka BP; uncalibrated 11,775 ± 30, 11,720 ± 50, and 11,615 ± 30 BP) and of brown bears immediately preceding (～14.5 cal. ka BP; 12,440 ± 35, 12,425 ± 30 BP) and following this time (～13 cal. ka BP; uncal. 11,100 ± 30 BP), suggesting niche partitioning to reduce competition among these species. We suggest that shifts in food availability or quality due to post-glacial vegetation and faunal changes were probably of primary importance in the arrival and the disappearance of giant short-faced bears on Vancouver Island. This study focuses on a key time period and geographic location that is useful in understanding Pleistocene extinctions in North America.     

Enzyme‐ and immunohistochemical aspects of skeletal muscle fibers in brown bear (Ursus arctos)

To further elucidate the pattern of MHC isoform expression in skeletal muscles of large mammals, in this study the skeletal muscles of brown bear, one of the largest mammalian predators with an extraordinary locomotor capacity, were analyzed. Fiber types in longissimus dorsi, triceps brachii caput longum, and rectus femoris muscles were determined according to the myofibrillar ATPase (mATPase) histochemistry and MHC isoform expression, revealed by a set of antibodies specific to MHC isoforms. The oxidative (SDH) and glycolytic enzyme (α‐GPDH) capacity of fibers was demonstrated as well. By mATPase histochemistry five fiber types, i.e., I, IIC, IIA, IIAX, IIX were distinguished. Analyzing the MHC isoform expression, we assume that MHC‐I, ‐IIa, and ‐IIx are expressed in the muscles of adolescent bears. MHC‐I isoform was expressed in Type‐I fibers and coexpressed with presumably ‐IIa isoform, in Type‐IIC fibers. Surprisingly, two antibodies specific to rat MHC‐IIa stained those fast fibers, that were histochemically and immunohistochemically classified as Type IIX. This assumption was additionally confirmed by complete absence of fiber staining with antibody specific to rat MHC‐IIb and all fast fiber staining with antibody that according to our experience recognizes MHC‐IIa and ‐IIx of rat. Furthermore, quite high‐oxidative capacity of all fast fiber types and their weak glycolytic capacity also imply for MHC‐IIa and ‐IIx isoform expression in fast fibers of bear. However, in adult, full‐grown animal, only MHC‐I and MHC‐IIa isoforms were expressed. The expression of only two fast isoforms in bear, like in many other large mammals (humans, cat, dog, goat, cattle, and horse) obviously meets the weight‐bearing and locomotor demands of these mammals. J. Morphol., 2009. © 2008 Wiley‐Liss, Inc.     

Ancient DNA analysis reveals divergence of the cave bear, Ursus spelaeus, and brown bear, Ursus arctos, lineages

The cave bear, Ursus spelaeus, represents one of the most frequently found paleontological remains from the Pleistocene in Europe. The species has always been confined to Europe and was contemporary with the brown bear, Ursus arctos. Relationships between the cave bear and the two lineages of brown bears defined in Europe, as well as the origins of the two species, remain controversial, mainly due to the wide morphological diversity of the fossil remains, which makes interpretation difficult [1, 2]. Sequence analysis of ancient DNA is a useful tool for resolving such problems because it provides an independent source of data [3]. We previously amplified a short DNA fragment of the mitochondrial DNA control region (mt control region) of a 40,000-year-old Ursus spelaeus sample [4]. In this paper, we describe the DNA analysis of two mtDNA regions, the control region and the cytochrome b gene. Control region sequences were obtained from ten samples of cave bears ranging from 130,000 to 20,000 years BP, and one particularly well-conserved sample gave a complete cyt b sequence. Our data demonstrate that cave bears split largely before the lineages of brown bears around 1.2 million years ago. Given its abundance, its wide distribution in space and time, and its large morphological diversity, the cave bear is a promising model for direct observation of the evolution of sequences throughout time, extinction periods, and the differentiation of populations shaped by climatic fluctuations during the Pleistocene.     

Integrating sign surveys and telemetry data for estimating brown bear (Ursus arctos) density in the Romanian Carpathians

Accurate population size estimates are important information for sustainable wildlife management. The Romanian Carpathians harbor the largest brown bear (Ursus arctos) population in Europe, yet current management relies on estimates of density that lack statistical oversight and ignore uncertainty deriving from track surveys. In this study, we investigate an alternative approach to estimate brown bear density using sign surveys along transects within a novel integration of occupancy models and home range methods. We performed repeated surveys along 2‐km segments of forest roads during three distinct seasons: spring 2011, fall‐winter 2011, and spring 2012, within three game management units and a Natura 2000 site. We estimated bears abundances along transects using the number of unique tracks observed per survey occasion via N‐mixture hierarchical models, which account for imperfect detection. To obtain brown bear densities, we combined these abundances with the effective sampling area of the transects, that is, estimated as a function of the median (± bootstrapped SE) of the core home range (5.58 ± 1.08 km²) based on telemetry data from 17 bears tracked for 1‐month periods overlapping our surveys windows. Our analyses yielded average brown bear densities (and 95% confidence intervals) for the three seasons of: 11.5 (7.8–15.3), 11.3 (7.4–15.2), and 12.4 (8.6–16.3) individuals/100 km². Across game management units, mean densities ranged between 7.5 and 14.8 individuals/100 km². Our method incorporates multiple sources of uncertainty (e.g., effective sampling area, imperfect detection) to estimate brown bear density, but the inference fundamentally relies on unmarked individuals only. While useful as a temporary approach to monitor brown bears, we urge implementing DNA capture–recapture methods regionally to inform brown bear management and recommend increasing resources for GPS collars to improve estimates of effective sampling area.     

Food habits and foraging of the brown bear Ursus arctos in central south Norway

Analysis of 134 excrement samples from the brown bear Ursus arctos L in boreal forests m central south Norway found volumes to be largest for Vaccinium spp, herbage Empetrum spp, and Formicidae The most important plant material in volume uptake were herbage, leaves, and mosses, closely followed by berries and seeds All together, plant material amounted to c 85% by volume, Formicidae to c 8%, and domestic sheep to c 7% These values were radically changed by applying correction factors for different nutritional value of food items The importance of large mammals then increased substantially, while the importance of herbage was reduced There was a drastic shift in food composition through the seasons Formicidae and flowers of deciduous trees were prevalent in spring, herbage in early summer, Vaccinium spp and sheep in late summer, and Vaccinium spp and Empetrum spp in autumn     

Microevolution of the mitochondrial DNA control region in the Japanese brown bear (Ursus arctos) population.

We investigated nucleotide sequences of the mitochondrial DNA control region to describe natural genetic variations and to assess the relationships between subpopulations of the brown bear Ursus arctos on Hokkaido Island, Japan. Using the polymerase chain reaction product-direct sequencing technique, partial sequences (about 930 bases) of the control region were determined for 56 brown bears sampled throughout Hokkaido Island. A sequence alignment revealed that the brown bear control region included a variable sequence on the 5' side and a repetitive region on the 3' side. Phylogenetic trees reconstructed from the 5' variable region (696-702 bases) exhibited 17 haplotypes, which were clustered into three groups (Clusters A, B, and C). The distribution of each group did not overlap with those of the others, and the three different areas were located in separate mountainous forests of Hokkaido Island. Furthermore, most of the phylogenetically close haplotypes within each group were distributed geographically close to each other. In addition, the 3' repetitive region (arrays of 10 bases) exhibited a much faster mutation rate than the 5' variable region, resulting in heteroplasmy. Such mitochondrial DNA divergence in each group could have occurred after the brown bears migrated from the continent to Hokkaido and became fixed in the different areas.     

Monitoring the effective population size of a brown bear (Ursus arctos) population using new single-sample approaches

The effective population size (N(e) ) could be the ideal parameter for monitoring populations of conservation concern as it conveniently summarizes both the evolutionary potential of the population and its sensitivity to genetic stochasticity. However, tracing its change through time is difficult in natural populations. We applied four new methods for estimating N(e) from a single sample of genotypes to trace temporal change in N(e) for bears in the Northern Dinaric Mountains. We genotyped 510 bears using 20 microsatellite loci and determined their age. The samples were organized into cohorts with regard to the year when the animals were born and yearly samples with age categories for every year when they were alive. We used the Estimator by Parentage Assignment (EPA) to directly estimate both N(e) and generation interval for each yearly sample. For cohorts, we estimated the effective number of breeders (N(b) ) using linkage disequilibrium, sibship assignment and approximate Bayesian computation methods and extrapolated these estimates to N(e) using the generation interval. The N(e) estimate by EPA is 276 (183-350 95% CI), meeting the inbreeding-avoidance criterion of N(e) > 50 but short of the long-term minimum viable population goal of N(e) > 500. The results obtained by the other methods are highly consistent with this result, and all indicate a rapid increase in N(e) probably in the late 1990s and early 2000s. The new single-sample approaches to the estimation of N(e) provide efficient means for including N(e) in monitoring frameworks and will be of great importance for future management and conservation.     

Nuclear DNA microsatellite analysis of genetic diversity and gene flow in the Scandinavian brown bear (Ursus arctos)

In the 1930s, the Scandinavian brown bear was close to extinction due to vigorous extermination programmes in Norway and Sweden. Increased protection of the brown bear in Scandinavia has resulted in the recovery of four subpopulations, which currently contain close to 1000 individuals. Effective conservation and management of the Scandinavian brown bear requires knowledge of the current levels of genetic diversity and gene flow among the four subpopulations. Earlier studies of mitochondrial DNA (mtDNA) diversity revealed extremely low levels of genetic variation, and population structure that grouped the three northern subpopulations in one genetic clade and the southernmost subpopulation in a second highly divergent clade. In this study, we extended the analysis of genetic diversity and gene flow in the Scandinavian brown bear using data from 19 nuclear DNA microsatellite loci. Results from the nuclear loci were strikingly different than the mtDNA results. Genetic diversity levels in the four subpopulations were equivalent to diversity levels in nonbottlenecked populations from North America, and significantly higher than levels in other bottlenecked and isolated brown bear populations. Gene flow levels between subpopulations ranged from low to moderate and were correlated with geographical distance. The substantial difference in results obtained using mtDNA and nuclear DNA markers stresses the importance of collecting data from both types of genetic markers before interpreting data and making recommendations for the conservation and management of natural populations. Based on the results from the mtDNA and nuclear DNA data sets, we propose one evolutionarily significant unit and four management units for the brown bear in Scandinavia.