Wound healing during hibernation by black bears (Ursus americanus) in the wild: elicitation of reduced scar formation

Even mildly hypothermic body or limb temperatures can retard healing processes in mammals. Despite this, we observed that hibernating American black bears (Ursus americanus Pallas, 1780) elicit profound abilities in mounting inflammatory responses to infection and/or foreign bodies. In addition, they resolve injuries during hibernation while maintaining mildly hypothermic states (30-35 °C) and without eating, drinking, urinating or defecating. We describe experimental studies on free-ranging bears that document their abilities to completely resolve cutaneous cuts and punctures incurred during or prior to hibernation. We induced small, full-thickness cutaneous wounds (biopsies or incisions) during early denning, and re-biopsied sites 2-3 months later (near the end of denning). Routine histological methods were used to characterize these skin samples. All biopsied sites with respect to secondary intention (open circular biopsies) and primary intention (sutured sites) healed, with evidence of initial eschar (scab) formation, completeness of healed epidermis and dermal layers, dyskeratosis (inclusion cysts), and abilities to produce hair follicles. These healing abilities of hibernating black bears are a clear survival advantage to animals injured before or during denning. Bears are known to have elevated levels of hibernation induction trigger (delta-opioid receptor agonist) and ursodeoxycholic acid (major bile acid within plasma, mostly conjugated with taurine) during hibernation, which may relate to these wound-healing abilities. Further research as to the underlying mechanisms of wound healing during hibernation could have applications in human medicine. Unique approaches may be found to improve healing for malnourished, hypothermic, diabetic and elderly patients or to reduce scarring associated with burns and traumatic injuries.     

Matrilinear hierarchy in the American black bear (Ursus americanus)

The American black bear (Ursus americanus) was long thought to be solitary and its social organization has not been well described. Here, we present new data on black bear social structure. The objectives of the study were to make detailed observations of the behavior of wild black bears to determine their social interactions and structure. We tested whether black bears interacted socially beyond mating and competing for resources, if black bears tracked relationships and interacted regularly even when resources were not limited, and whether the social structure of a population of black bears was based on a matrilinear hierarchy. We collected data by direct observation of bears from 1993 to 2014. Observations of 1210 social interactions at a provisioning site indicated that females compete and form matrilinear hierarchies. Dominant bears established a hierarchy for food, control of space, and control of younger bears. Post interaction scent marking took place, which suggested that dominant females were conditioning subordinates to their scent marks. Affiliative behavior occurred between related and unrelated bears and helped to establish the social structure of the bear community. Based on our data, human–bear conflicts can be reduced by behavioral modifications by humans when they encounter bears. Knowledge of bear behavior and the matrilinear hierarchy provide a basis for non-lethal management of bears that find themselves in a bear–human conflict situation.     

Blindness in a wild American black bear cub (Ursus americanus)

An approximately six‐month‐old wild American black bear (Ursus americanus) was found wandering in Saskatchewan and was presented to the Veterinary Medical Centre of the Western College of Veterinary Medicine for apparent blindness. Clinical examination confirmed an inability to navigate a photopic maze, bilateral tapetal hyper‐reflectivity, fundi devoid of retinal vessels, and small pale optic nerve papillae. Single‐flash electroretinography revealed A and B‐wave amplitudes of approximately 40 and 140 microvolts, respectively, in both eyes. Histologic abnormalities included bilateral optic papillary mineralization and bilateral segmental optic nerve degeneration, with occasional intralesional lymphocytes confirmed with immunohistochemistry for CD3+. There was also bilateral multifocal retinal dysplasia, gliosis, lymphocytic retinitis, a complete lack of retinal blood vessels, an intravitreal vascular membrane, and a mild lymphocytic–plasmacytic uveitis with small pre‐iridal cellular membranes. The presence of a positive ERG in a blind bear with numerous retinal ganglion cells and degenerative changes in the optic nerve are most consistent with vision loss due to optic nerve injury, which given the young age of the bear likely occurred during ocular development. The presence of ocular inflammation suggests this injury resulted from an inflammatory/infectious process. The etiology could not be determined. Hepatic concentrations of vitamin A were within the normal reference range for domestic species. Pan‐herpesvirus PCR and immunohistochemistry for canine distemper virus and Toxoplasma gondii were negative, although this does not rule out these or other infectious etiologies. This represents the first case report of neonatal or congenital ocular abnormalities in an ursid species.     

Sex steroid and prolactin profiles in male American black bears (Ursus americanus) during denning

Serum sex steroid and prolactin profiles were examined in the male American black bear, Ursus americanus during denning. Sera collected in December and the following March from 8 denning male black bears in Minnesota, U.S.A. were assayed for testosterone, estradiol-17 beta and prolactin. Eight bears were confirmed to be the denning mode based on a serum urea to creatinine ratio less than 10. Serum testosterone concentrations tended to increase from December to the subsequent March whereas serum estradiol-17 beta concentrations tended to decrease during this period. There were few changes in serum prolactin concentrations between December and March. These findings suggest that spermatogenesis and testicular steroidogenesis initiated during denning may be influenced by changes in serum sex steroid concentrations in the American black bear.     

Antebrachial fractures in four captive polar bears (Ursus maritimus)

OBJECTIVE: To identify common risk factors for antebrachial fractures of captive polar bears and to evaluate outcome after fracture repair. STUDY DESIGN: Retrospective study. ANIMALS: Four captive polar bears. METHODS: United States zoological collections were surveyed to determine the prevalence of fractures in captive polar bears. Medical records of captive polar bears that had antebrachial fractures were reviewed for signalment, history, physical and radiographic findings, fracture management, postoperative care, and outcome. Serum samples from healthy bears and bears with antebrachial fractures were assayed for 25-hydroxyvitamin D (25-OHD) concentrations. RESULTS: Nineteen fractures (12 polar bears) occurred from 1974 to 2002; 12 fractures involved the antebrachium. Management of 4 antebrachial fractures was reviewed; 3 were repaired by internal fixation and 1 by external coaptation. Fractures healed and bears were returned to exhibit on average 3 months postfracture. Of 11 serum samples assayed for 25-OHD concentrations, 6 were below normal, 1 was low normal and 4 were within normal reference intervals. The 7 bears with subnormal or low normal values were housed in 2 zoos. Subnormal vitamin D concentrations were identified in 2 of 3 bears with fractures. CONCLUSIONS: Fracture disease is not uncommon in captive polar bears. Additional research is necessary to explore the role of nutrition in polar bear fracture disease. CLINICAL RELEVANCE: Internal fixation of antebrachial fractures is feasible and reasonably well tolerated in captive polar bears.     

Blood lipid concentrations and lipoprotein patterns in captive and wild American black bears (Ursus americanus)

OBJECTIVE: To compare blood lipid concentrations and lipoprotein patterns for captive and wild American black bears (Ursus americanus). ANIMALS: 7 captive and 9 wild adult (> or = 4 years old) black bears. PROCEDURE: Blood was collected from 2 groups of captive black bears (groups A and B) and 1 group of wild black bears (group C). Blood triglyceride (TG) and cholesterol concentrations were compared among groups. Plasma lipoproteins were isolated by use of a self-generating gradient of iodixanol, and lipoprotein patterns were compared between groups A and B. RESULTS: Captive bears (mean +/- SD, 187.8 +/- 44.4 kg) weighed significantly more than wild bears (mean, 104.8 +/- 41.4 kg), but mean body weight did not differ between groups A and B. Mean blood TG concentrations for groups B (216.8 +/- 16.0 mg/dL) and C (190.7 +/- 34.0 mg/dL) were significantly higher than that of group A (103.9 +/- 25.3 mg/dL). Mean blood cholesterol concentration was also significantly higher for group B (227.8 +/- 8.2 mg/dL) than for groups A (171.7 +/- 35.5 mg/dL) or C (190.8 +/- 26.8 mg/dL). Mean very-low-density lipoprotein TG and low-density lipoprotein cholesterol concentrations were 2- and 3-fold higher, respectively, for group B, compared with concentrations for group A. CONCLUSIONS AND CLINICAL RELEVANCE: Blood lipid concentrations vary significantly among populations of black bears. Plasma lipoprotein patterns of captive bears differed significantly between colonies and may have reflected differences in diet or management practices.     

Mobility of the forearm skeleton in the Asiatic black (Ursus thibetanus), brown (U. arctos) and polar (U. maritimus) bears

In several primates and carnivores, pronation/supination angles of the forearm skeleton were examined, and it is thought that a larger angle is useful to acquire dexterous behaviors in feeding and/or life style, including climbing. In this study, the pronation/supination angles in Asiatic black, brown and polar bears were nondestructively examined. These specimens were classified as adult or non-adult. Three or four carcasses of each group of Asiatic black and brown bears were used for CT analysis, whereas only one adult polar bear was used. The forearms were positioned within the gantry of a CT scanner in both maximally supinated and pronated states. Extracted cross-sectional CT images of two positions were superimposed by overlapping the outlines of each ulna. The centroids of the radii were detected, and then the centroid of each radius and the midpoint of a line which connects between both ends of the surface of each radius facing the ulna, were connected by lines to measure the angle of rotation as an index of pronation/supination. In adult brown and polar bears, the angles were smaller as compared with the other groups (Asiatic black and non-adult brown bears). Asiatic black and non-adult brown bears can climb trees, whereas adult brown bears and polar bears cannot. This suggests that the pronation/supination angle is related to arboreal activity in Ursidae.     

Laryngeal squamous cell carcinoma in a North American black bear (Ursus americanus).

A 10-yr-old female black bear (Ursus americanus) at the North Carolina Zoological Park presented with a 2-wk history of intermittent coughing and stertor. The animal was anesthetized for examination, and at intubation, a large mass associated with the tonsils and larynx was observed obstructing the airway. Cardiopulmonary collapse occurred during the procedure, and efforts at resuscitation were ineffective. At necropsy, an 8 cm times 6 cm times 5 cm mass was present in the larynx, infiltrating the epiglottis and obstructing the airway. Regional lymph nodes were mildly enlarged and contained tan inspissated substance. Histologically, the neoplasm was composed of nests of keratinizing squamous epithelial cells with evidence of lymphatic invasion and metastases to regional lymph nodes.     

Microscopic and molecular identification of hemotropic mycoplasmas in South American coatis (Nasua nasua)

Hemoplasmas were detected in two apparently healthy captive South American coatis (Nasua nasua) from southern Brazil during an investigation for vector-borne pathogens. Blood was subjected to packed cell volume (PCV) determination, a commercial real-time PCR panel for the detection of Anaplasma spp., Babesia spp., Bartonella spp., Hepatozoon spp., Leishmania spp., Mycoplasma haemofelis, ‘Candidatus Mycoplasma turicensis’, ‘Candidatus Mycoplasma haemominutum’, Neorickettsia risticii, Rickettsia rickettsii and Leptospira spp., and a pan-hemoplasma conventional PCR assay. PCV was normal, but both coatis tested positive for hemoplasmas and negative for all the remaining pathogens tested. Using different techniques for microscopy (light, confocal or SEM), structures compatible with hemoplasmas were identified. Sequencing of the 16S rRNA gene identified an organism resembling Mycoplasma haemofelis and another hemotropic Mycoplasma sp., with a sequence identity of 96.8% to a Mycoplasma sp. previously detected in capybaras.     

Sarcoptic mange and Pelodera dermatitis in an American black bear (Ursus americanus)

An adult female free-ranging American black bear (Ursus americanus) was presented in poor body condition, with advanced skin disease. Skin changes included hair loss, lichenification, crusting, and focal erosions. Skin scrapings and histopathology identified two distinct parasitic conditions that were contributing to this animal's dermatitis. Large numbers of larvae, nymphs, and adults of Sarcoptes scabiei were present in the superficial epidermis, and nematodes consistent with Pelodera strongyloides were abundant within the hair follicles. This appears to be the first reported case of Pelodera dermatitis in a bear species, adding a new differential agent to the list for dermatitis in bears. The sarcoptic mange and poor body condition of this bear may have been contributing factors to the development of the opportunistic Pelodera infestation.     

Helicobacter spp. from captive bottlenose dolphins (Tursiops spp.) and polar bears (Ursus maritimus)

The gastric fluid of six bottlenose dolphins and the faeces of four polar bears from the same oceanarium were examined for the presence of Helicobacter. As detected by PCR, all dolphins and 8/12 samples collected from polar bears were positive for Helicobacter. Novel sequence types were identified in samples collected from these animals of which several were unique to either the dolphins or the polar bears. At least one sequence type was, however, detected in both animal taxa. In addition, a sequence type from a dolphin shared a 98.2-100% identity to sequences from other Helicobacter species from harp seals, sea otters and sea lions. This study reports on the occurrence of novel Helicobacter sequence types in polar bears and dolphins and demonstrates the broad-host range of some species within these animals.     

Magnetic resonance imaging diagnosis of intervertebral disc disease and myelomalacia in an American black bear (Ursus americanus)

A 23-yr-old black bear (Ursus americanus) was examined because of paralysis of unknown duration. The precise onset of clinical signs was unknown as a result of seasonal torpor. The bear was immobilized and transported to a university veterinary teaching hospital for further evaluation and treatment. Radiography revealed increased mineral opacity and ventral bridging across vertebral segments T8-11. Magnetic resonance imaging demonstrated dorsal and ventral compression of the spinal cord at T8-9. Given the bear's advanced age, the unknown duration of spinal cord compression, unknown presence of deep pain perception, and thus an unknown prognosis for surgical success, euthanasia was elected. Postmortem examination revealed severe spondylosis deformans from T7 to L3 and dorsal extradural extruded disc material in the area of T8-9. Histopathology demonstrated the dorsal horns of the spinal cord at T9 were replaced by foamy macrophages extending into the dorsal and lateral funiculi of the white matter compatible with focal, severe, chronic myelomalacia. This is the first report of intervertebral disc disease and myelomalacia diagnosed using MRI in a large carnivore.     

Molecular detection of filarial nematode parasites in Japanese black bears (Ursus thibetanus japonicus) from Iwate Prefecture, Japan

This study aimed to detect filarial parasites in blood samples of Japanese black bears (Ursus thibetanus japonicus) collected from Iwate Prefecture, Japan. Positive amplicons were obtained from 26 out of 30 samples by nested PCR targeting 18S ribosomal RNA gene and first internal transcribed spacer regions. DNA sequences of Mansonella sp. close to M. ozzardi and Dirofilaria sp. were detected for eight and 11 positive amplicons, respectively. Co-infection was detected for the remaining seven amplicons. Dirofilaria sp. was identified as D. ursi by further genetic analysis of 5S ribosomal RNA gene sequence. The results of this study will contribute to further investigations of Japanese black bears for monitoring their risk as a reservoir of possible zoonotic filarial parasites.     

Thyroid cystadenoma, colloid goiter, and hypothyroidism in an American black bear (Ursus americanus).

A 178-kg, 14-yr-old captive female American black bear (Ursus americanus) was examined because of lethargy, inappetance, obesity, and alopecia. Serum chemistry and complete blood count values were within normal limits. Based on serum levels for total thyroxine (T4), free T4 by equilibrium dialysis (fT4ED), and canine thyroid-stimulating hormone concentrations, using assays validated for domestic dogs, hypothyroidism was diagnosed presumptively, and therapy with levothyroxine sodium (0.022 mg/kg p.o. b.i.d.) was initiated. Haircoat, body weight, appetite, and activity level improved within 30 days. The levothyroxine dose was decreased twice (to 0.018 mg/kg p.o. b.i.d. and then to 0.011 mg/kg p.o. b.i.d.) during the course of treatment based on monitoring of serum T4 and fT4ED concentrations. After euthanasia for severe refractory lameness, postmortem examination revealed bilateral thyroid lobe enlargement and a fluid-filled cyst within the right lobe. Histologically, colloid goiter was present in both lobes, and a follicular cystadenoma had replaced one third of the cranial pole of the right lobe. The goiter and cystadenoma likely contributed to the hypothyroid condition in this bear and fT4ED was a more sensitive indicator of hypothyroidism than was T4. The recommended canine dosage of levothyroxine may be too high for the treatment of hypothyroidism in American black bears; 0.011 mg/kg p.o. b.i.d. may be a more appropriate dosage.     

Occurrence of hemotropic mycoplasmas in non-human primates (Alouatta caraya,Sapajus nigritus and Callithrix jacchus) of southern Brazil

Hemoplasmas, the erythrocyte-associated mycoplasmas, have been detected in several primates, causing mostly subclinical infection. This study aimed to determine the prevalence of hemoplasma infection in captive and free-ranging monkeys from southern Brazil, as well as factors and hematological abnormalities associated with infection. Blood samples from 40 non-human primates (NHP) were tested for hemoplasmas and coinfections. An overall of 10/40 (25.0%) NHP tested positive for hemoplasmas using PCR-based assays, including 9/14 (64.3%) black howler monkeys (Alouatta caraya) and 1/24 (4.2%) black-horned capuchin (Sapajus nigritus). Infection was not statistically associated with anemia, but wild-born monkeys and male black howler monkeys were more likely to be positive when compared with captive-born animals and female black howler monkeys, respectively. The sequences from the black howler monkey hemoplasma were similar (94% identity) to the squirrel monkey hemoplasma (“Candidatus Mycoplasma kahanei”) and were phylogenetically located in a different cluster when compared to the human hemoplasma (“Candidatus Mycoplasma haemohominis”).     

Electroejaculation and semen cryopreservation of free-ranging Japanese black bears (Ursus thibetanus japonicus)

The Japanese black bear (Ursus thibetanus japonicus) is endangered for extinction in some areas of Japan, and semen collection and cryopreservation are an important means to preserve genetic resources. The aim of this study was to characterize and cryopreserve semen of free-ranging Japanese black bears. Semen was collected by electroejaculation procedure from 4 free-ranging Japanese black bears at the capture point in the field. Ejaculates containing motile sperm were recovered from all of the animals and ejaculate volume, total sperm count, % motility (percentage of motile spermatozoa), % viability (percentage of spermatozoa that excluded eosin) and % abnormal morphology (range (mean)) were 0.65-2.20 (1.51) ml, 99-1082 (490) x 10(6), 5-100 (31), 42-97 (66) and 20-87 (53), respectively. Three of the 4 ejaculates were diluted with an egg yolk-TRIS-citrate-glucose extender and cryopreserved in liquid nitrogen. Motile spermatozoa were observed after freezing and thawing in all cases. This study showed that electroejaculation would be a useful method for collecting semen from free-ranging Japanese black bears and that at least motile spermatozoa would be obtained by freezing the thus collected electroejaculates.     

Characteristics of a hepatozoonosis in lungs of Japanese black bears (Ursus thibetanus japonicus)

In a survey of pathologic agents in the wild animals of central Japan, we found a Hepatozoon sp. in the lungs of Japanese black bears in Fukui, Shiga, and Gifu Prefectures, Japan. Histopathologic examination of organs and tissues from the 18 bears inspected showed hepatozoonosis in all. Immature and mature meronts were found in all lobules and between alveoli, with a few found between pleura and in connective tissue. In the lungs, inflammatory cells were not found around meronts, merozoites, or tubercles made of macro-phages including zoites, but inflammatory cells were found around degenerating cells, zoites, and tubercles. A Hepatozoon sp. has not been reported as being detected in bears of any species before.     

Characteristics of frozen-thawed spermatozoa cryopreserved with different concentrations of glycerol in captive Japanese black bears (Ursus thibetanus japonicus)

Seven mature Japanese black bears were used as semen donors, and a total of 7 semen samples collected from the animals by the electroejaculation method were cryopreserved in liquid nitrogen. Egg yolk-TRIS-citrate-glucose extender was used, and the effects of different final concentrations of glycerol, at 4-12% (v/v), on frozen-thawed spermatozoa were examined. No significant difference was observed in percent motility or percent abnormal morphology of frozen-thawed spermatozoa among the different glycerol concentrations. Percent viability and percent intact acrosomes of spermatozoa cryopreserved with 4 and 6% glycerol were significantly higher than those with 10 and 12% glycerol. These results suggest that a suitable glycerol concentration for freezing Japanese black bear semen within the range tested would be 4-6%.     

Immobilization of Japanese black bears (Ursus thibetanus japonicus) with tiletamine hydrochloride and zolazepam hydrochloride

The effect of anesthetizing with a 1:1 combination of tiletamine hydrochloride and zolazepam hydrochloride (TZ) was evaluated in 75 Japanese black bears. TZ was administered to 43 captive and 11 wild, 8 captives and 13 hibernating captive bears at the doses of approximately 9.0 mg/kg (usual dosage), 18.0 mg/kg (high dosage) and 5.0 mg/kg (low dosage), respectively. Sufficient anesthetic effects were achieved in all bears, and rectal temperatures, heart rates and respiratory rates did not change significantly during an hour handling. Complete blood cell examinations showed no abnormal data. A combination of TZ would be an efficient and safe drug for chemical immobilization of Japanese black bears.     

Haplosporidium nelsoni (MSX) in Eastern Oyster Populations of North Carolina

Abstract

Populations of the eastern oyster Crassostrea virginica from 36 locations off the coast of North Carolina were examined cytologically in the fall of 1989 for Haplosporidium nelsoni (MSX). The pathogen was found in oysters from 11 (31%) of the sites. Systemic Perkinsus marinus infections were noted in oysters from 26 (72%) of the locations.