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A comparative chromosome banding analysis of the Ursidae and their relationship to other carnivores.

Trypsin G-banded karyotypes of eight species of Ursidae were prepared from retrovirus-transformed skin fibroblast cultures. The banding patterns of all bears are highly conserved, even though their diploid numbers range from 42 to 72. A comprehensive analysis of the homologous banding patterns within the Ursidae and with a hypothesized ancestral carnivore karyotype permitted the reconstruction of three significant chromosomal reorganization events that occurred during the evolution of the modern ursids. The first was a multichromosomal fissioning away from the biarmed (2n = 44) primitive carnivore karyotype, leading to six species of the Ursinae subfamily (2n = 78). The second was a comprehensive chromosome fusion in the lineage that led to the Ailuropodinae (giant panda) subfamily (2n = 44). The third event was a second, independent, but less extensive, centromeric fusion occurring in the line that led to the Tremarctinae (spectacled bear) subfamily (2n = 52). Ursidae karyotypes are not only highly conserved within the family but also exhibit extensive chromosome banding homology with other carnivore families.

Animals↗

Ursodeoxycholic acid in the Ursidae: biliary bile acids of bears, pandas, and related carnivores.

The biliary bile acid composition of gallbladder bile obtained from six species of bears (Ursidae), the Giant panda, the Red panda, and 11 related carnivores were determined by reversed phase liquid chromatography and gas chromatography-mass spectrometry. Bile acids were conjugated solely with taurine (in N-acyl linkage) in all species. Ursodeoxycholic acid (3 alpha, 7 beta-dihydroxy-5 beta-cholan-24-oic acid) was present in all Ursidae, averaging 1-39% of biliary bile acids depending on the species; it was not detected or present as a trace constituent (< 0.5%) in all other species, including the Giant panda. Ursodeoxycholic acid was present in 73 of 75 American Black bears, and its proportion averaged 34% (range 0-62%). Ursodeoxycholic acid averaged 17% of biliary bile acids in the Polar bear (n = 4) and 18% in the Brown bear (n = 6). Lower proportions (1-8%) were present in the Sun bear (n = 2), Ceylon Sloth bear (n = 1), and the Spectacled bear (n = 1). Bile of all species contained taurine-conjugated chenodeoxycholic acid and cholic acid. In some related carnivores, deoxycholic acid, the 7-dehydroxylation product of cholic acid, was also present. To determine whether the 7 beta hydroxy group of ursodeoxycholic acid was formed by hepatic or bacterial enzymes, bile acids were determined in hepatic bile obtained from bears with chronic biliary fistulae. Fistula bile samples contained ursodeoxycholic acid, chenodeoxycholic acid, and a trace amount of cholic acid, all as taurine conjugates, indicating that ursodeoxycholic acid is a primary bile acid formed in the liver in Ursidae.(ABSTRACT TRUNCATED AT 250 WORDS)

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Rapid radiation events in the family Ursidae indicated by likelihood phylogenetic estimation from multiple fragments of mtDNA.

The bear family (Ursidae) presents a number of phylogenetic ambiguities as the evolutionary relationships of the six youngest members (ursine bears) are largely unresolved. Recent mitochondrial DNA analyses have produced conflicting results with respect to the phylogeny of ursine bears. In an attempt to resolve these issues, we obtained 1916 nucleotides of mitochondrial DNA sequence data from six gene segments for all eight bear species and conducted maximum likelihood and maximum parsimony analyses on all fragments separately and combined. All six single-region gene trees gave different phylogenetic estimates; however, only for control region data was this significantly incongruent with the results from the combined data. The optimal phylogeny for the combined data set suggests that the giant panda is most basal followed by the spectacled bear. The sloth bear is the basal ursine bear, and there is weak support for a sister taxon relationship of the American and Asiatic black bears. The sun bear is sister taxon to the youngest clade containing brown bears and polar bears. Statistical analyses of alternate hypotheses revealed a lack of strong support for many of the relationships. We suggest that the difficulties surrounding the resolution of the evolutionary relationships of the Ursidae are linked to the existence of sequential rapid radiation events in bear evolution. Thus, unresolved branching orders during these time periods may represent an accurate representation of the evolutionary history of bear species.

Animals↗

Comparative anatomy of the cardiac foramen ovale in cats (Felidae), dogs (Canidae), bears (Ursidae) and hyaenas (Hyaenidae).

The structure of the foramen ovale from 16 species representing 4 carnivore families, the Felidae, Canidae, Ursidae and Hyaenidae, was studied using the scanning electron microscope. The Felidae were represented by 9 domestic cat fetuses (Felis catus), 2 snow leopard neonates (Uncia uncia), an ocelot neonate (Leopardus pardalis), 2 lion neonates (Panthera leo), a panther neonate (Panthera pardus) and 3 tigers (Neofelis tigris), comprising 2 fetuses and a neonate. The Canidae were represented by a golden jackal neonate (Canis aureus), a newborn wolf (Canis lupus), 8 domestic dog fetuses (Canis familiaris), 3 red fox neonates (Vulpes vulpes) and a dhole neonate (Cuon alpinus). The Ursidae were represented by a brown bear neonate (Ursus arctos), a day-old grizzly bear cub (Ursus arctos horribilis), a polar bear neonate (Ursus maritimus), and 2 additional bear fetuses (species unknown). The Hyaenidae were represented by a striped hyaena neonate (Hyaena hyaena). In each species, the foramen ovale, when viewed from the terminal part of the caudal vena cava, had the appearance of a short tunnel. A thin fold of tissue, the developed remains of the embryonic septum primum, extended from the distal end of the caudal vena cava for a variable distance into the lumen of the left atrium and contributed towards the 'tunnel' appearance in all specimens. It constituted a large proportion of the tube, and its distal end was straight-edged. There was fibrous material underlying the endothelium of the flap, the apparent morphology of which suggested that it comprised cardiac muscle.(ABSTRACT TRUNCATED AT 250 WORDS)

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Occurrence and structure of epipharyngeal pouches in bears (Ursidae).

The infrequent mention of epipharyngeal pouches occurring in some species of bears indicates the scarcity of morphological and functional knowledge about these structures. In order to provide precise morphological data on the structure of these remarkable formations and to verify their taxonomic utility, the pharyngeal regions of 1 spectacled bear and 3 brown bears were examined. All these individuals possessed epipharyngeal pouches, which are tubular, blind-ending outpouchings of the caudodorsal pharyngeal wall equipped with respiratory epithelium and a thick layer of elastic fibres. While the spectacled bear and Ursus arctos syriacus possessed a single pouch on the caudodorsal wall of the nasopharynx, in Ursus arctos and Ursus arctos beringianus 2 unequally sized pouches were present. Two additional sacs of smaller size, representing outpouchings of the lateral pharyngeal wall, occurred in the spectacled bear. These findings prove epipharyngeal pouches to be constant and unique morphological features of the family Ursidae, the anatomical features suggesting involvement in the respiratory system most probably in important aspects of ursid phonation. This is the first description of epipharyngeal pouches in the spectacled bear.

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An empirical evaluation of genetic distance statistics using microsatellite data from bear (Ursidae) populations.

A large microsatellite data set from three species of bear (Ursidae) was used to empirically test the performance of six genetic distance measures in resolving relationships at a variety of scales ranging from adjacent areas in a continuous distribution to species that diverged several million years ago. At the finest scale, while some distance measures performed extremely well, statistics developed specifically to accommodate the mutational processes of microsatellites performed relatively poorly, presumably because of the relatively higher variance of these statistics. At the other extreme, no statistic was able to resolve the close sister relationship of polar bears and brown bears from more distantly related pairs of species. This failure is most likely due to constraints on allele distributions at microsatellite loci. At intermediate scales, both within continuous distributions and in comparisons to insular populations of late Pleistocene origin, it was not possible to define the point where linearity was lost for each of the statistics, except that it is clearly lost after relatively short periods of independent evolution. All of the statistics were affected by the amount of genetic diversity within the populations being compared, significantly complicating the interpretation of genetic distance data.

Alleles↗

Comparative genomics: tracking chromosome evolution in the family ursidae using reciprocal chromosome painting.

The Ursidae family includes eight species, the karyotype of which diverges somewhat, in both chromosome number and morphology, from that of other families in the order Carnivora. The combination of consensus molecular phylogeny and high-resolution trypsin G-banded karyotype analysis has suggested that ancestral chromosomal fissions and at least two fusion events are associated with the development of the different ursid species. Here, we revisit this hypothesis by hybridizing reciprocal chromosome painting probes derived from the giant panda (Ailuropoda melanoleuca), domestic cat (Felis catus), and man (Homo sapiens) to representative bear species karyotypes. Comparative analysis of the different chromosome segment homologies allowed reconstruction of the genomic composition of a putative ancestral bear karyotype based upon the recognition of 39 chromosome segments defined by painting as the smallest conserved evolutionary unit segments (pSCEUS) among these species. The different pSCEUS combinations occurring among modern bear species support and extend the postulated sequence of chromosomal rearrangements and provide a framework to propose patterns of genome reorganization among carnivores and other mammal radiations.

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[The occurrence of tumors in large bears (Ursidae)--a literature review and six case descriptions].

Histological findings on two Malayan sun bears and four sloth bears show that malignant neoplasms play an important role in tropic bears. Further, most of the tumors originated from the hepatic and biliary tract. Our results were compared with other investigations on zoo animals during the last 70 years revealing that malignant neoplasms are the most common ones in bears of the family Ursidae. Accordingly to our results, sloth and Malayan sun bears seem to have a disposition to develop malignant neoplasms of the hepatic and biliary tract, but within other species only polar bears seem to suffer predominantly from such neoplasia. The reason for this phenomenon could be an alimentary intake of carcinogens. Furthermore, Malayan sun bears show very often neoplasms of the thyroid gland as it is also observed in other carnivora.

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Phylogenetic relationships of bears (the Ursidae) inferred from mitochondrial DNA sequences.

The phylogenetic relationships among some bear species are still open questions. We present here mitochondrial DNA sequences of D-loop region, cytochrome b, 12S rRNA, tRNA(Pro), and tRNA(Thr) genes from all bear species and the giant panda. A series of evolutionary trees with concordant topology has been derived based on the combined data set of all of the mitochondrial DNA sequences, which may have resolved the evolutionary relationships of all bear species: the ancestor of the spectacled bear diverged first, followed by the sloth bear; the brown bear and polar bear are sister taxa relative to the Asiatic black bear; the closest relative of the American black bear is the sun bear. Primers for forensic identification of the giant panda and bears are proposed. Analysis of these data, in combination with data from primates and antelopes, suggests that relative substitutional rates between different mitochondrial DNA regions may vary greatly among different taxa of the vertebrates.

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Phylogeography of brown bears (Ursus arctos) of Alaska and paraphyly within the Ursidae.

Complete nucleotide sequences of the mitochondrial cytochrome b, tRNA(prolime), and tRNA(threonine) genes were described for 166 brown bears (Ursus arctos) from 10 geographic regions of Alaska to describe natural genetic variation, construct a molecular phylogeny, and evaluate classical taxonomies. DNA sequences of brown bears were compared to homologous sequences of the polar bear (maritimus) and of the sun bear (Helarctos malayanus), which was used as an outgroup. Parsimony and neighbor-joining methods each produced essentially identical phylogenetic trees that suggest two distinct clades of mtDNA for brown bears in Alaska: one composed only of bears that now reside on some of the islands of southeastern Alaska and the other which includes bears from all other regions of Alaska. The very close relationship of the polar bear to brown bears of the islands of southeastern Alaska as previously reported by us and the paraphyletic association of polar bears to brown bears reported by others have been reaffirmed with this much larger data set. A weak correlation is suggested between types of mtDNA and habitat preference by brown bears in Alaska. Our mtDNA data support some, but not all, of the currently designated subspecies of brown bears whose descriptions have been based essentially on morphology.

Alaska↗

A phylogeny of the bears (Ursidae) inferred from complete sequences of three mitochondrial genes.

Complete sequences of DNA are described for the cytochrome b tRNA(Thr) and tRNA(Pro) genes of mitochondria of four extant species of ursids and compared to sequences of four other species of ursids previously studied by us. Phylogenetic analyses indicate that the giant panda and the spectacled bear are the basal taxa of the ursid radiation. The ursines, a group which includes the sun bear, sloth bear, American black bear, Asiatic black bear, brown bear, and polar bear, experienced a rapid radiation during the mid Pliocene to early Pleistocene. The Asiatic black bear and American black bear are sister taxa. The brown bear and polar bear are the most recently derived of the ursines, with the polar bear originating from within a clade of brown bears during the Pleistocene. This paraphyletic association suggests that the rate of morphological evolution may be accelerated relative to that of molecular evolution when a new ecological niche is occupied. Calibration of the corrected average number of nucleotide differences per site with the fossil record indicates that transitions at third positions of codons in the ursid cytochrome b gene occur at a rate of approximately 6% per million years, which is considerably slower than comparable values reported for other species of mammal.

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Basic haematological values in carnivores--I. The Canidae, the Hyaenidae and the Ursidae.

Basic haematological values for 32 animals of four carnivore species are reported. In six adult wolves (Canis lupus) the mean values estimated for the erythrocyte count 7.48 X 10(12)/l, haematocrit 0.465/l, haemoglobin 172.0 g/l and leukocyte counts 7.33 X 10(9)/l are given. For five young wolves, these parameters were markedly decreased, only the white cell count was raised. In 14 hunting dogs (Lycaon pictus) the mean values estimated are: erythrocyte count 9.15 x 10(12)/l haematocrit 0.435/l, haemoglobin 179.2 g/l and leukocytes 12.95 X 10(9)/l. In six striped hyaenas (Hyaena hyaena) the mean estimated values are: erythrocyte count 8.11 X 10(12)/l, haemoatocrit 0.445/l, haemoglobin 178.0 g/l and leukocytes 13.95 X 10(9)/l. Only individual values for the reported parameters are given in the Asiatic black bear (Selenarctos thibetanus). All results are compared with values derived from the literature for animals under investigation and for the domestic dog (Canis familiaris).

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Phylogeny of the bears (Ursidae) based on nuclear and mitochondrial genes.

The taxomic classification and phylogenetic relationships within the bear family remain argumentative subjects in recent years. Prior investigation has been concentrated on the application of different mitochondrial (mt) sequence data, herein we employ two nuclear single-copy gene segments, the partial exon 1 from gene encoding interphotoreceptor retinoid binding protein (IRBP) and the complete intron 1 from transthyretin (TTR) gene, in conjunction with previously published mt data, to clarify these enigmatic problems. The combined analyses of nuclear IRBP and TTR datasets not only corroborated prior hypotheses, positioning the spectacled bear most basally and grouping the brown and polar bear together but also provided new insights into the bear phylogeny, suggesting the sister-taxa association of sloth bear and sun bear with strong support. Analyses based on combination of nuclear and mt genes differed from nuclear analysis in recognizing the sloth bears as the earliest diverging species among the subfamily ursine representatives while the exact placement of the sun bear did not resolved. Asiatic and American black bears clustered as sister group in all analyses with moderate levels of bootstrap support and high posterior probabilities. Comparisons between the nuclear and mtDNA findings suggested that our combined nuclear dataset have the resolving power comparable to mtDNA dataset for the phylogenetic interpretation of the bear family. As can be seen from present study, the unanimous phylogeny for this recently derived family was still not produced and additional independent genetic markers were in need.

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Transrectal ultrasonographic examination of the female urogenital tract in nonpregnant and pregnant captive bears (Ursidae).

The development of reliable methods for monitoring the reproductive cycle of bears is needed to optimise breeding management in captivity. The aim of the present study was to develop non-invasive procedures for obtaining basic data on the reproductive status of nonpregnant and pregnant bears. Eight female captive bears (five Ursus arctos arctos, two Tremarctos ornatus and one Melursus ursinus) were examined using transrectal adaptor ultrasonographic imaging. Plasma concentrations of progesterone and gestagens in faeces were determined simultaneously. The ultrasonographic appearance of the female urogenital tract was assessed during the nonbreeding season and verified in four adult bears post mortem. Pregnant bears were examined twice, as the reproductive physiology of bears is characterized by delayed implantation and pseudopregnancy. During embryonic diapause there was no difference between pseudopregnant and pregnant bears both by ultrasonographic imaging and endocrinological data. After implantation (early December) the pregnancies were visualised by ultrasonographic imaging, whereas plasma concentrations of progesterone and gestagens in faeces had not yet increased. In conclusion, both transrectal ultrasonographic imaging and gestagen monitoring in faeces are efficient methods for obtaining important data on reproduction in bears.

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