A retrospective study of dental conditions of captive brown bears (Ursus arctos spp.) compared with free-ranging Alaskan grizzlies (Ursus arctos horribilis).

The teeth of 63 skulls of brown bears (Ursus arctos spp.) that had lived in the Bernese bear pit between 1850 and 1995 were examined and radiographed for occlusion, loss of teeth, calcified dental plaque and calculus, attrition, and caries. Dental pathology and periodontal status were evaluated with respect to age and location in the dentition. Further techniques such as computed tomography, microhardness testing, histologic sectioning, and scanning electron microscopy were used in selected cases to verify the macroscopic and radiographic diagnoses. Fourteen skulls originating from free-ranging wild Alaskan brown bears (Ursus arctos horribilis) were examined as a control group. For zoo bears over 10 yr old, severe enamel and dentinal attrition has been observed in canine teeth, with exposed pulp and proximal lesions in molar teeth. Deposits of calcified dental calculus were found in various locations and increased with age. A much lower degree of calculus deposition was found in the Alaskan grizzlies, whose average caries frequency was higher but with large individual variation and different locations in canine teeth and occlusal lesions in molar and premolar teeth. Radiographic evaluation of perialveolar osteolytic processes revealed a greater frequency of apical and combined apical-marginal lesions of the alveoli in zoo bears over 10 yr old. Stereotypical behavior like cage chewing is a suspected cause of canine tooth and secondary alveolar lesions, whereas a nutritionally inappropriate diet and inadequate opportunities for tooth-cleaning activities are responsible for the lack of natural cleaning and the extensive calculus formation that results. The methods used in this study are useful in assessing and comparing the dental health status of free-ranging bears, and those living in zoological gardens, by evaluating anesthetized bears or bear skull collections representing both groups.     

Anesthesia of polar bears (Ursus maritimus) with zolazepam-tiletamine, medetomidine-ketamine, and medetomidine-zolazepam-tiletamine.

A 1:1 combination (by weight) of zolazepam and tiletamine is the drug of choice for anesthetizing polar bears (Ursus maritimus), but recovery time is prolonged when additional doses are administered. Recoveries may last 24 hr and may threaten the health of the bears. We compared the anesthetic effects of zolazepam-tiletamine (ZT) with those of medetomidine-ketamine (MK) and medetomidine-zolazepam-tiletamine (MZT) in 93 free-ranging polar bears. The MZT combination was administered in smaller dose and volume, resulted in more rapid, safer, and more predictable induction, provided more reliable anesthesia, and was safely reversed with atipamezole. Frequent occurrence of sudden recoveries during anesthesia with MK limited our use of this combination. MK and MZT sometimes caused apnea and bradycardia initially and hyperthermia at increased ambient temperatures. Hypoxemia occurred transiently with all combinations. When anesthesia with ZT and MK exceeded 1 hr, frequent necessary top-up doses caused irregular physiologic function. ZT is recommended for short duration anesthesia (< or = 1 hr), but MZT is better for anesthesia of longer duration and under circumstances where reversibility is desirable.     

Carfentanil citrate used as an oral anesthetic agent for brown bears (Ursus arctos).

Carfentanil citrate was given orally to five adult brown bears (Ursus arctos) on 14 separate occasions during the winter and summer to determine effective anesthetic dosages and how season may alter these dosages. Lower blood urea nitrogen:creatinine ratios, depressed appetite, and decreased activity levels in the winter versus summer were reflective of different metabolic states, even though bears were not hibernating in the winter. Doses of carfentanil citrate between 6.0 and 15.2 microg/kg were mixed with 5-10 ml of honey, which the bears licked voluntarily from a spoon. During each anesthetization, respiratory and heart rates, hemoglobin saturation, temperature, electrocardiogram, blood gas values, and level of consciousness were monitored and utilized to determine effective dosages. Mean (+/- SE) dose requirements in the winter were 7.6 +/- 0.4 microg/kg, whereas a greater mean dose of 12.7 +/- 0.5 microg/kg was required in the summer (P < 0.05). After ingestion began, sternal recumbency occurred in an average of 7.5 min (range: 4-11 min), and full restraint and safe handling was achieved in 21 min (range: 8-40 min). At the end of each procedure, naltrexone was given as the reversal agent at a ratio of 100 mg naltrexone per 1 mg carfentanil, with 25% of the dose given i.v. or i.m. and 75% given s.c. Mean reversal time was 6 min after injection of naltrexone (range: 4-9 min). Rapid induction and recovery times and ease of oral administration make carfentanil citrate an effective anesthetic agent for use in brown bears. However, hypoventilation and respiratory acidosis were noted in all bears, and oxygen insufflation is recommended.     

Spinal spondylosis and acute intervertebral disc prolapse in a European brown bear (Ursus arctos arctos)

A 22-year-old male European brown bear (Ursus arctos arctos) was presented to the Onderstepoort Veterinary Academic Hospital after an acute onset of hind limb paralysis 4 days earlier. Previous radiographs revealed marked degenerative joint disease of the stifles, tarsi and digits. The clinical findings were consistent with acute disc prolapse. Lateral radiographs of the entire vertebral column were made as well as ventrodorsal pelvic radiographs. The latter were within normal limits. The vertebral column revealed multiple lesions consistent with chronic and acute disc herniations. Lateral compression of the caudal lumbar nerve roots could not be ruled out. Owing to multiple significant findings of the vertebral column and the poor prognosis for full recovery after surgery, the bear was euthanased. The diagnosis of an acute disc prolapse and multiple chronic disc herniations was confirmed on necropsy.     

Occurrence of Baylisascaris transfuga in wild populations of European brown bears (Ursus arctos) as identified by a new PCR method

The European brown bear (Ursus arctos) is a species present in limited areas of Europe and several small populations are considered endangered. This species can be affected by a range of parasites. In particular, the genus Baylisascaris is an emerging parasite of wild animals which can cause severe larva migrans syndrome in aberrant hosts, which include 100 species of birds, mammals and also humans. Baylisascaris transfuga is the species reported from bears, and with the exception of a few laboratory trials, little is known about the capacity of this species to infect other animals. Furthermore, the identification of this species has traditionally been based on light microscopy, using either morphology of the adults at necropsy or detection of the eggs in faeces, which are methods limited by the experience and the efforts of the observer. The current work was aimed at developing a specific PCR to detect the parasite directly from faecal samples of naturally infected brown bears in the field, without the need for previous flotation. Using eggs and adults of B. transfuga collected in Croatia, we first developed a PCR to detect a portion of the second internal transcribed spacer region (ITS-2) of ribosomal DNA and then applied it to bear faecal samples spiked with a known number of B. transfuga eggs. We show here for the first time that this method allows the detection of a minimum of two Baylisascaris eggs in 25mg of faecal material, thus it demonstrates a high diagnostic capacity that could be applied to evaluate the prevalence of the parasite in faecal samples from wild populations of brown bears.     

Serum progesterone concentrations using P-EIA kit in captive and free-ranging Hokkaido brown bears, Ursus arctos yesoensis.

Serum progesterone (P) concentrations using P-EIA kit (Ovucheck, Cambridge Life Science Co., Ltd.) were examined in 8 captive and 7 free-ranging female Hokkaido brown bears (Ursus arctos yesoensis). The intra- and inter-assay coefficients of variation were 8.9%, 12.6% and 16.6%, 22.7%, respectively, based on 2 serum samples. There was a significant correlation between EIA and radioimmunoassay results based on 64 serum samples (r = 0.725; p less than 0.01). Serum P concentrations were examined in 5 pregnant, 2 solitary non-pregnant bears and a lactating non-pregnant bear in captivity. Annual changes of P levels in pregnant bears were observed as a small elevation during the mating season (May-June), a re-elevation in September-October and a sharp elevation in November-December. The sharp elevation was suspected to reflect changes when implantation occurred. Annual changes of P levels in solitary non-pregnant bears were similar to those in pregnant bears. An annual change of P levels in a lactating non-pregnant bear maintained levels under 5 ng/ml. Two of 7 free-ranging bears exhibited P levels over 1 ng/ml and the birth of cubs was confirmed in the following year in 1 of the 2 bears. P concentrations of other free-ranging bears exhibited less than 1 ng/ml, and these bears were considered to be non-pregnant. It was concluded that P-EIA kit was available for measuring P concentrations in Hokkaido brown bears.     

Immobilization of sun bears (Helarctos malayanus) with medetomidine-zolazepam-tiletamine.

A mixture of medetomidine (50.0 microg/kg, i.m.) and zolazepam-tiletamine (2.0 mg/kg, i.m.) effectively immobilized 16 sun bears (Helarctos malayanus) for more than 1 hr with good myorelaxation and minimal effects on cardiorespiratory performance during 22 immobilizations. All bears were immobilized once, except for six individuals that were immobilized twice. Atipamezole (250 microg/kg, i.v.) effectively reversed medetomidine-induced sedation and reduced recovery time significantly. Respiratory rates of immobilized bears did not change significantly over time. Rectal body temperature and heart rate decreased significantly after 10 min of immobilization. Hematologic and serum biochemical parameters did not change significantly within 30 min of induction.     

In vitro maturation of oocytes derived from the brown bear (Ursus arctos)

This study was conducted to determine whether meiotic maturation could be induced in ovarian oocytes from the American brown bear (Ursus arctos), a model for gamete "rescue" techniques for endangered ursids. The bears were euthanized, and their ovaries were transported to the laboratory within 4 h. The mean ovarian size was 2.4 x 1.8 cm (range: 2.0-3.3 x 1.5-2.2 cm). The ovaries obtained from the 2 brown bears yielded 97 oocytes (48.5/female), and 88 (90.7%) of them were morphologically classified as normal quality. Oocytes were in vitro matured at 38.5 C in 5% CO2 for 24 or 48 h in TCM-199 supplemented with 10% FBS, 1 microg/ml estradiol-17beta, and 10 microg/ml FSH. In Exp. 1, morphologic evaluation of matured oocytes was conducted by measuring the diameters of oocytes with a zona pellucida (ZP) or cytoplasm without a ZP. In Exp. 2, activation was induced by applying two 20 microsec DC pulses of 2.0 kV/cm delivered by an Electro Cell Fusion Generator. The activated oocytes were cultured in TCM-199 containing 2 mM of 6-dimethylaminopurine for 4 h, in Charles Rosenkrans (CR) 1 for 3 days and the in CR2 for another 4 days. The diameters of the matured bear oocytes with a ZP and with cytoplasm without a ZP (161.8 +/- 6.0 and 135.3 +/- 7.5 microm, respectively) were significantly (P<0.05) larger than those of bovine oocytes (150.7 +/- 4.9 and 118.7 +/- 7.5 microm). The maturation rates of the bear oocytes were 17.6 and 59.4% at 24 and 48 h of in vitro maturation, the percentage of activated oocytes that developed to the 2 or 4-cell stage was 31.6%; however, no blastocysts were observed. These results indicate that bear oocytes can develop to metaphase II in an in vitro culture system and that activated oocytes can develop to the 2 or 4-cell stages.     

Use of fluoxetine to treat stereotypical pacing behavior in a brown bear (Ursus arctos)

A 12-year-old, male, brown bear (Ursus arctos) named Abdi exhibited stereotypical pacing behavior. He was kept as a pet for 10 years by local villagers, then he was rescued and taken to the Karacabey Bear Sanctuary in July 2001. His physical condition was extremely poor. Because he had never lived with other bears, he was terrified and refused to integrate with them. After 6 months he was healthy physically but observations by the keepers indicated that he was pacing all day. He was treated with fluoxetine (0.62 mg/kg orally every 24 hours) for 6 months, and had been observed totally for 18 months. After stereotypy ceased completely, he was transferred to the large naturalistic enclosure and did not show any stereotypical behavior during the subsequent observation period (1 year). He was treated successfully using a combination of fluoxetine, the provisioning of extra space, and the addition of novel stimulation in a naturalistic enclosure. The space and stimulation could be sufficient to mimic the therapeutic effect of the pharmacological therapy in the long term.     

Brown bear (Ursus arctos arctos) femoral head and neck excision.

A 30-yr-old untamed European female brown bear (Ursus arctos arctos) with a craniodorsal luxation of the right femoral head and bilateral degenerative joint disease of the coxofemoral joint had a femoral head and neck excision following unsatisfactory conservative medical therapy. The bear was injected with zolazepam-tiletamine, and anesthesia was induced with i.v. thiopental and maintained with isoflurane in oxygen via endotracheal tube. A lumbosacral epidural injection of medetomidine-bupivacaine provided additional analgesia. Slight initial cardiorespiratory depression was counteracted with fluid and inotropic drug administration and ventilatory assistance. The bear's gluteal muscle anatomy differs from that of the dog. Recovery was uneventful. The bear was confined indoors for 6 wk and was able to ambulate normally within 6 mo.     

Changes in sexual behavior and fecal steroid hormone concentrations during the breeding season in female Hokkaido brown bears (Ursus arctos yesoensis) under captive condition

The longitudinal changes in fecal steroid hormone concentrations and sexual behavior in 2 mated/pregnant and 3 non-mated female Hokkaido brown bears were investigated during the breeding season. Behavioral estrus (standing) lasted for 14 and 32 days in the mated females and for 25 to 36 days in the non-mated females. In non-mated females, sexual behavior, such as female-female mounting and masturbation, was observed for several days before and after the estrous period. In mated females, mean fecal estradiol-17beta concentrations were higher in the estrous period than in the post-estrous period, while fecal progesterone concentrations were higher in the post-estrous period than in the estrous period. The similar trends of steroid hormone changes were observed in the non-mated females.     

Optimization of glycerol concentration and freezing rate in the cryopreservation of ejaculate from brown bear (Ursus arctos)

In order to establish a semen bank for the endangered Cantabrian brown bear, we tested five glycerol concentrations and three freezing rates for electroejaculated semen. Electroejaculation was performed on nine males. Semen was diluted in TES-Tris-Fructose (20% egg yolk, 2% EDTA, 1% Equex) with 2%, 4%, 6%, 8% or 10% glycerol and frozen at -10, -20 or -40°C/min. Before and after cryopreservation, samples were analysed for motility (CASA), viability and acrosomal status (flow cytometry). Pre-freezing results showed that glycerol concentration had no significant effect on total motility or progressive motility, but it significantly decreased VCL, ALH, viability and acrosomal status (p < 0.05). After thawing, sperm motility was higher at extender with 4%, 6% and 8% glycerol, but only at 4% and 6% glycerol for viability and acrosomal status. For 4% and 6% glycerol, freezing rates did not have significant effects. The curve fitting gave an estimate of the optimal glycerol concentration, with all the optimal values for every parameter between 6% and 7% glycerol falling. We propose using 6% glycerol and a freezing velocity of -20°C/min for freezing brown bear ejaculated spermatozoa.     

Cryopreservation of the semen collected by electroejaculation from the Hokkaido brown bear (Ursus arctos yesoensis)

Four adult Hokkaido brown bears were used as semen donors, and semen characteristics were examined before freezing and after thawing. A total of 10 electroejaculates were diluted with Tris-egg yolk extender and cooled to 4 degrees C over 90 min. Spermatozoa were equilibrated with 4.7% glycerol for 80 min. Semen packed in 0.25 ml plastic straws were frozen with liquid nitrogen vapor. Percentages (mean +/- SD) of motile and live sperm were 96+/-2 and 86.5+/-7.2% before freezing, and 43+/-5 and 67.4+/-3.9% after thawing, respectively. Although the number of progressively motile sperm after thawing varied among samples (1.8+/-1.2 x 10(8) cells/ejaculate), frozen semen in the present study might serve for artificial insemination.     

Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for mtDNA typing in hokkaido brown bear (Ursus arctos yesoensis)

To develop an easy method of typing of mitochondrial DNA (mtDNA) in Hokkaido brown bears (Ursus arctos yesoensis), the PCR-RFLP technique was improved using four restriction enzymes: Mbo 1, Cfr 13 I, TspE 1, and Fok 1. This approach identified seven groups of mtDNA haplotypes, HB1/2/5-7, HB 3, HB4, HB8/9, HB10/11, HB12 and HB13 from 102 brown bears of northern, central and eastern Hokkaido.     

Effect of the change of social environment on the behavior of a captive brown bear (Ursus arctos)

We observed the behavior of a captive sterilized male brown bear before and after the death of his female sibling to investigate the effect of the change of social environment on his behavior and welfare. Observations were carried out by continuous recording during daylight for 6 wk prior to hibernation when the bear was kept with the sibling, and they were repeated 2 years later when the bear was alone (total observation time is 108 h). Feeding, moving, and the total amount of time spent resting (including sleeping and alert inactive) were not affected by the change of social environment. However, when the bear was alone, the percentage of time he spent alert inactive almost trebled (pair, 17.7 ± 3.3%; alone, 48.5 ± 5.5%; P < 0.001), and the time spent sleeping was less than one-third (pair, 51.1 ± 6.1%; alone, 14.2 ± 5.0%; P < 0.001) than when the female was present. The bear spent most of his sleeping time in lateral lying posture (a posture probably associated with rapid eyes movement sleep). The percentage of time dedicated to this posture was significantly reduced after the death of his sibling (pair, 35.9 ± 7.4%; alone, 15.0 ± 5.4%; P < 0.05), whereas the percentage of time spent in quadrupedal posture increased (pair, 17.1 ± 5.4%; alone, 37.8 ± 7.8; P < 0.05). One of the possible reasons for these changes may be an increased risk perception of the bear after the death of his sibling. Our results highlight the importance of social environment and of its changes, which should be carefully considered to maintain captive bears in good welfare conditions.     

Efficiency of fecal steroid hormone measurement for assessing reproductive function in the Hokkaido brown bear (Ursus arctos yesoensis)

The present study aimed to establish simple systems for measuring fecal steroid hormones in order to monitor the reproductive profiles of captive Hokkaido brown bears. The efficiency of fecal sample processing at the steps of dehydration and extraction and the correlation between steroid concentrations in matched fecal and blood samples were studied. Then, monthly changes in fecal estradiol-17 beta and progesterone in female bears, and testosterone in male bears were examined. The procedure was finalized as follows. Fecal samples were dried at 100 degrees C for 3 hr and extracted with diethyl ether. The diethyl ether in the extracts was evaporated and residues were reconstituted in ethanol for the assays. Hormone concentrations were quantified using enzyme immunoassays. Concentrations of progesterone and testosterone in fecal and plasma samples were correlated in the systems. The changes in fecal progesterone and testosterone concentrations were similar to those in serum concentrations of bears as reported previously. In contrast, fecal estradiol concentrations did not correlate with plasma levels probably because of the time lag in excretion. However, the changes in estradiol-17 beta concentrations in feces in the present study were similar to those reported in serum. In conclusion, fecal progesterone and testosterone assay systems appear practical for monitoring ovarian and testicular activities without immobilization, though methodological improvements and further validation may be required. For the fecal estradiol-17 beta assay, there is a need to solve the problem of excretion time lag before the system can be used in the study of reproductive physiology.     

Comparative anatomy of the radial sesamoid bone in the polar bear (Ursus maritimus), the brown bear (Ursus arctos) and the giant panda (Ailuropoda melanoleuca).

Since we have clarified the manipulation mechanism using the radial sesamoid (RS) in the giant panda (Ailuropoda melanoleuca), our aim in this study is to examine the position, shape and function of the RS morphologically, and to observe the attachment to the RS of the M. abductor pollicis longus and the M. opponens pollicis in the other Ursidae species. So, we focused on the carpus and manus of the polar bear (Ursus maritimus) and the brown bear (Ursus arctos) in this study. The RS was tightly articulated to the radial carpal, and could not adduct-abduct independently of the radial carpal. The M. abductor pollicis longus tendon and the M. opponens pollicis belly were attached to the RS, independently. In the polar bear, the deep concave and the flat surface were confirmed in attachment area for these two muscles. The morphological relationship between the RS and the M. abductor pollicis longus and the M. opponens pollicis in the two species of bears were essentially consistent with that in the giant panda. It also demonstrated that the manipulation mechanism of the giant panda has been completely based on the functional relationship between the small RS, and the M. abductor pollicis longus and the M. opponens pollicis in Ursidae species.     

Humeral Chondrosarcoma in a Hokkaido Brown Bear (Ursus arctos yesoensis)

Humeral chondrosarcoma was found in an 18-year-old male Hokkaido brown bear (Ursus arctos yesoensis). Necropsy revealed a large firm mass under the left superficial pectoral muscle at the axillary region. The mass involved the left shoulder joint and peripheral muscles, and connected to the head of the humerus with osteolysis. Histopathologically, the mass was composed of irregularly shaped myxomatous to cartilaginous tumor lobules. The tumor cell showed moderate nuclear atypia with a relatively high mitotic index, especially in the edges of the myxomatous lobules. The tumor cells were positively immunostained with vimentin and S-100 protein. Based on these findings, the tumors were diagnosed as chondrosarcoma. Metastases were found in the left axillary lymph node, lungs, liver and kidney.