Noninvasive monitoring of stress hormone levels in a female steller sea lion (Eumetopias jubatus) pup undergoing rehabilitation.

Steller sea lions (Eumetopias jubatus) rarely strand in areas monitored by humans, and there is little published data on the diseases, parasites, nutritional state, and stress levels of Steller sea lions in the wild. In May 2002, a female Steller sea lion pup (EJS-02-01) was sighted separated from her mother after strong storms in Southeast Alaska. After 5 days of observations, EJS-02-01 was transferred to the Alaska SeaLife Center (ASLC) in Seward, Alaska. During 11 mo of rehabilitation at ASLC, body weight was monitored and opportunistic fecal samples (n = 86) were analyzed for corticosterone concentrations. Fecal corticosterone concentrations ranged from 15 to 3,805 ng/ g for EJS-02-01. Peak corticosterone values reflected responses to acute stressors during rehabilitation. EJS-02-01 was successfully released at Gran Point, Alaska, in April 2003. Fecal corticosterone assay monitoring provided a valuable tool to monitor various stressors and is useful in monitoring long-term situations like rehabilitation.     

Characterization of ocular gland morphology and tear composition of pinnipeds

Objective The importance of tear film integrity to ocular health in terrestrial mammals is well established, however, in marine mammals, the role of the tear film in protection of the ocular surface is not known. In an effort to better understand the function of tears in maintaining health of the marine mammal eye surface, we examined ocular glands of the California sea lion and began to characterize the biochemical nature of the tear film of pinnipeds. Procedures Glands dissected from California sea lion eyelids and adnexa were examined for gross morphology, sectioned for microscopic analysis, and stained with hematoxylin and eosin. The tear film was examined using interferometry. Tears were collected from humans and pinnipeds for the analysis of protein and carbohydrate content. Results The sea lion has sebaceous glands in the lid, but these glands are different in size and orientation compared with typical meibomian glands of terrestrial mammals. Two other accessory ocular glands located dorsotemporally and medially appeared to be identical in morphology, with tubulo‐acinar morphology. An outer lipid layer on the ocular surface of the sea lion was not detected using interferometry, consistent with the absence of typical meibomian glands. Similar to human tears, the tears of pinnipeds contain several proteins but the ratio of carbohydrate to protein was greater than that in human tears. Conclusions Our findings indicate that the ocular gland architecture and biochemical nature of the tear film of pinnipeds have evolved to adapt to the challenges of an aquatic environment.     

Evaluation of the repair process of cartilage defects of the equine third carpal bone with and without subchondral bone perforation

To determine the effect of subchondral bone drilling (forage) on the cartilage repair process after injury has occurred, a cartilage defect (1 cm in diameter) was created on the radial facet of the proximal surface of each third carpal bone in 6 adult horses. In one of the third carpal bones (right or left thoracic limb) of each horse, a 1-cm cartilage defect was created, and 5 holes (1 mm in diameter and 10 mm deep) were drilled through the subchondral bone into the cancellous bone. In the other thoracic limb, an identical defect was created, but not drilled. Analyses of cell numbers and types in the synovia and the mucin precipitate quality were done before, at 1 week after, and 3 weeks after surgical manipulation was done and showed no significant difference between the joint environment of drilled carpi and those of nondrilled carpi. At 21 weeks after surgical manipulation was done, each joint was examined radiographically, macroscopically, and microscopically to compare the condition of the joints and the state of repair of the cartilage in each defect. The amount of surface of the defect covered by the dense fibrous and fibrocartilagenous repair tissue and the thickness of the repair tissue were significantly greater (P less than 0.05 and P less than 0.01, respectively) in the drilled carpal bones. In addition, the attachment of the repair tissue to underlying chondro-osseous tissue was better in the drilled carpal bones. Fibrocartilage was resurfacing the drilled defects, whereas only fibrous tissue was present in the nondrilled defects.(ABSTRACT TRUNCATED AT 250 WORDS)     

Healing of full-thickness cartilage compared with full-thickness cartilage and subchondral bone defects in the equine third carpal bone.

The effect of lesion depth on the quality of third carpal bone cartilage repair was examined. A 1-cm diameter articular defect penetrating the calcified cartilage in one limb and the subchondral bone plate in the opposite limb was created in the radial facet of the third carpal bones. Clinical and xeroradiographic examinations were performed every 4 weeks until 4 months (3 horses) and 6 months (3 horses) after surgery. The synovial membrane, non-opposing articular surfaces and articular defects were examined grossly, histologically and histochemically. Grossly, deeper defects contained thicker, whiter tissue, but both joints contained generalised degenerative changes. Defects extending through calcified cartilage were filled deeply by fibrocartilage and superficially by fibrous connective tissue. Defects extending through subchondral bone were consistently filled with hyaline-like cartilage in the depths of the lesion, fibrocartilage in the intermediate layer and fibrous connective tissue superficially. The results indicate that subchondral bone is the source of hyaline-like cartilage repair tissue and suggest that quality of healing of cartilage defects may be improved by penetrating the subchondral bone plate. It also appears that the synovitis associated with the procedure must be controlled before the procedure can be advocated for treatment of clinical cases.     

New marine calicivirus serotype infective for swine

A new serotype of calicivirus was isolated from California sea lions (Zalophus californianus) with severe vesicular disease. Neutralizing antibodies were found in 27 of 82 (32.9%) serum samples from California sea lions and in 15 of 146 (10.3%) serum samples from Steller sea lions (Eumetopias jubatus) tested. The seropositive animals were widely dispersed along the margins of the eastern Pacific basin, from the Bering Sea to the Santa Barbara Channel. Seropositive samples were found from as early as 1976 through the present time. This new calicivirus serotype, San Miguel sea lion virus type 13, was inoculated into weaned pigs, resulting in induction of severe vesicular disease, which spread to all pigs, including uninoculated pen contacts. Virus was continually shed by most of the pigs throughout the 2-week duration of the experiment.     

Magnetic resonance imaging‐detected adaptation and pathology in the distal condyles of the third metacarpus,associated with lateral condylar fracture in Thoroughbred racehorses

Reasons for performing the study: Lateral condylar (LC) fractures of the third metacarpus (McIII) are a common reason for euthanasia in racehorses, and may be the result of repetitive overloading or cumulative pathological change. Magnetic resonance imaging (MRI) allows monitoring of bone and cartilage to detect pathological and adaptive changes that may be precursors of fracture. Objectives: To describe bone and cartilage MRI features in the distal condyles of McIII of Thoroughbred racehorses, with and without condylar fracture. Hypotheses: 1) A greater degree of bone and cartilage adaptation or pathology will be seen in fractured McIIIs compared with their respective contralateral McIIIs. 2) Contralateral McIIIs will have a greater degree of bone and cartilage adaptation or pathology than McIIIs from control horses that did not sustain a LC fracture. Methods: The McIIIs from 96 horses subjected to euthanasia at racecourses were divided into 3 groups: Group 1: nonfractured bones from horses without LC fracture; Group 2: nonfractured bones from horses with unilateral LC fracture; and Group 3: fractured bones from horses with unilateral LC fracture. The MR images were examined and graded for bone and cartilage changes. Results: Nine percent of Group 1 (n = 9) and 11% of Group 2 bones (n = 5) had incomplete LC fractures. Focal palmar necrosis was most frequently detected in bones from Group 1 (12%) compared with Groups 2 (9%) and 3 (4%). The prevalence of bone and/or cartilage abnormalities tended to increase from Group 1 to Group 2 to Group 3. Conclusions: Magnetic resonance imaging is able to detect cartilage and bone changes that may be associated with LC fracture. There was no significant difference in bone/cartilage changes between bones from Groups 1 and 2, despite increased pathology in Group 2 bones. Potential relevance: Periodic monitoring of bone and/or cartilage changes in distal McIII of Thoroughbred racehorses may help to prevent catastrophic LC fractures.     

Osteocartilaginous metaplasia in the right atrial myocardium of healthy adult sheep.

Among 172 hearts from clinically normal adult sheep aged 1.5 to 7 years evaluated for the presence of cartilage and/or bone in the right atrial myocardium, 3.49% (6/172) had palpable evidence of osteocartilaginous foci. An additional 8% prevalence was estimated based on radiographs of hearts that contained < or =1 mm foci of nonpalpable, radiographically dense bone. Microscopically, the nodules in the atria were characterized by mature lamellar bone enclosing adipose tissue, with occasional new bone formation by endochondral ossification. No degenerative changes were evident in the affected atrial myocardium, suggesting that these lesions were clinically insignificant background changes.     

An explanation of a groove found on the nasal process of the equine incisive bone

Thirteen fresh equine heads were dissected in an attempt to determine the cause of a groove frequently found on the dorsal border and medial side of the nasal process of the incisive bone. This groove appeared on both sides in 40 out of 44 equine skulls. The sulcus seems to be caused by the combined action of the lateralis nasi muscle and the medial accessory cartilage of the nose (cartilago nasalis accessoria medialis). Other sulci found on the nasal process of the bone may be explained as impressions caused by nerves.     

Congenital chondrodysplastic dwarfism with dyshematopoiesis in a holstein calf

A holstein calf with congenital chondrodysplastic dwarfism was histopathologically examined. The head of the calf was relatively flat giving a dog-like appearance with its short nose and sloping forehead. Limb bones were dumbbell-like with short diaphysis and hypertrophied metaphyses. Bone marrow was pale, whitish and fatty. In the metaphyseal plates most of chondrocytes were pyknotic with swollen and ghost-like cytoplasm, and were irregularly arranged. Column of calcified cartilage were poorly formed losing comb-like structure. Bone marrow was ischemic with poor hematopoiesis and was moderately replaced by adipose tissue. In articular cartilage, most of chondrocytes were degenerated with ghost-like cytoplasm. Many cartilage canals and occasional bone marrow-like structure were formed. The characteristics lesions of the calf were chondrodysplasia and dyshematopoiesis.     

A NEWLY RECOGNIZED PATTERN OF CANINE OSTEOCHONDROMATOSIS

Clinicopathological findings are described for a unique skeletal pattern of osteochondromatosis (syn. multiple osteochondromas) in three crossbred littermate dogs. Multiple nonpainful bilaterally symmetric skeletal lesions arose from focal semiannular and annular areas of periosteal thickening on the cortical surface of the metaphyses and diaphyses of long bones. Flat bones of the skull were spared. In radiographs osteochondromas in different stages of evolution were apparent. Grossly there were smoothly contoured, nodular subperiosteal masses of dense fibrocartilaginous and osseous tissue that were continuous with and overlying apparently normal but thin trabeculae of cancellous bone. Cortical compacta was attenuated or absent beneath exostoses. In histologic sections of the cartilage-capped exostoses a hyperplastic periosteum formed a focal semiannular or annular cap of proliferative hyaline cartilage tissue that underwent endochondral ossification and replacement by cancellous bone at its base. Intertrabecular spaces in the cancellous bone of exostoses were filled with fibro-osseous and hematopoietic tissue. A hereditary origin was suspected for the osteochondromas. Origin of some osteochondromas as semiannular or annular perturbations of the perichondrial ring of metaphyseal physes of long bones likely contributed to limb shortening and a pattern of billaterally symmetric angular limb deformities of all limbs as occurs in some children affected by osteochondromatosis.     

Current prevalence of adult Uncinaria spp. in northern fur seal (Callorhinus ursinus) and California sea lion (Zalophus californianus) pups on San Miguel Island, California, with notes on the biology of these hookworms

A prevalence survey for hookworms (Uncinaria spp.) was done in northern fur seal (Callorhinus ursinus) and California sea lion (Zalophus californianus) pups on San Miguel Island, CA, in 2000. Intestines of dead pups were examined for adult hookworms in July. These parasites were found in 95% of 20 fur seal pups and 100% of 31 sea lion pups. The number of hookworms varied from 4 to 2142 (mean = 760) in fur seal pups and from 20 to 2634 (mean = 612) in sea lion pups. A direct relationship was evident between body condition and number of hookworms in the pups; that is, pups in poor condition had fewer hookworms than those in good condition. There was a decline in the number of hookworms in sea lion pups in 2000 compared to collections in 1996. Eggs of Uncinaria spp. were found in rectal feces (collected in late September and early October) of none of 35 (0%) live fur seal pups and 41 of 48 (85%) live sea lion pups. Packed cell volume values, determined for most of the same live pups, were essentially normal for C. ursinus but were much lower than normal for most Z. californianus. Hookworm larvae were not found in blubber of fur seal and sea lion pups or in rookery sand in July. Rookery sand, positive for live hookworm larvae when put in a refrigerator, was negative at removal 2.5 years later. The average number of eggs in utero of female hookworms was 285 for three specimens from a fur seal pup and 281 from three specimens from a sea lion pup. One hookworm larva was recovered from milk stripped from the teats of a stranded Z. californianus female at The Marine Mammal Center, Sausalito, CA.     

Effects of clustered drill holes on the breaking strength of the equine third metacarpal bone

The breaking strength (stress at failure) of equine third metacarpal bones, with and without clustered drill holes, was determined in vitro. Paired ossa metacarpalia II-IV of 39 horses (n = 39) between 2 and 7 years old were tested in palmarodorsal 3-point bending. Four treatments were compared. Clustered 2.7- or 3.5-mm drill holes, in a 4- or 7-hole pattern, were made in the dorsal cortex of the distal diaphysis of the left third metacarpal bone. Undrilled right third metacarpi were used as controls. Bones with clustered drill holes failed by an oblique fracture through 1 or more drill holes, whereas undrilled bones failed with a middiaphyseal transverse fracture. Clustered drill holes acted as a stress concentrator and significantly (P less than 0.05) decreased the stress required for failure. However, differences in breaking strength between treatment groups were not significant (P greater than 0.05).     

Pulsed Carbon Dioxide Laser for Cartilage Vaporization and Subchondral Bone Perforation in Horses Part II: Morphologic and Histochemical Reactions

A pulsed carbon dioxide laser was used to vaporize articular cartilage in four horses, and perforate the cartilage and subchondral bone in four horses. Both intercarpal joints were examined arthroscopically and either a 1 cm cartilage crater or a series of holes was created in the third carpal bone of one joint. The contralateral carpus served as a control. After euthanasia at week 8, the treated and control joints were examined for gross changes, and samples of cartilage and subchondral bone, synovial membrane, and peripheral lymph nodes were examined histologically. Depletion of cartilage matrix glycosaminoglycan was assessed by safranin-O histochemical staining of the laser site and adjacent cartilage. Cartilage removal by laser vaporization resulted in rapid regrowth, with fibrous and fibrovascular tissue and occasional regions of fibrocartilage at week 8. The subchondral bone, synovial membrane, and draining lymph nodes appeared essentially unaffected by the laser cartilage vaporization procedure. Conversely, carbon dioxide laser drilling of subchondral bone resulted in poor penetration, extensive areas of thermal necrosis of bone, and significant secondary damage to the apposing articular surface of the radial carpal bone.     

Anatomy of the splint bones and their surrounding area particularly in consideration of fascial attachments

This study concerns the vestigial metacarpal and metatarsal bones, so called splint bones. The soft tissues, which stabilize the splint bones were carefully examined. The palmar metacarpal/plantar metatarsal fascia, the metacarpal/metatarsal interosseous ligaments and the fibres, which exit distally from the splint bone, were examined in depth. Cadaveric distal limbs were collected from 5 slaughtered or euthanized horses. The legs were examined both, microscopically and macroscopically. The palmar metacarpal/plantar metatarsal fascia is a very stable structure, which connects the splint bone with its environment. The connection between splint and McIII/MtIII, established through the metacarpal/ metatarsal interosseous ligaments, showed impressive differences between specimens examined. Macroscopic examination of the soft tissue in the distal region showed a band structure, which originates from the distal end of the splint bone and extends in dorsal and distal direction towards the proximal sesamoid bone.     

EX VIVO MAGNETIC RESONANCE IMAGING OF THE DISTAL ROW OF EQUINE CARPAL BONES: ASSESSMENT OF BONE SCLEROSIS AND CARTILAGE DAMAGE

The distal row of carpal bones (C2, C3, and C4) from eight left intercarpal joints--four from Standardbred Trotters and four from Swedish Warmblood horses--were used to assess the potential of magnetic resonance (MR) imaging to detect cartilage and bone lesions. The joints used in the study were classified by macroscopic and radiographic examinations as having normal, mild, moderate, or severe articular cartilage lesions and bone sclerosis. Those classifications correlated well with the appearance of the MR images. Bone sclerosis in the MR images was observed as regions of decreased signal intensity. Upon quantitative analysis of the MR images there was a significant difference (p < 0.0001) in the MR signal intensity from areas where radiographic bone sclerosis was observed compared to areas of radiographic nonsclerotic bone. In addition, the MR images were used to pilot the location of histology slices through areas of interest that were then examined microscopically; hence, the lesions found from the MR imaging examination were verified microscopically. It was concluded that cartilage lesions and cartilage loss are related to the sclerotic state of the underlying bone. The MR protocols developed in this study were applied on five intact cadaveric carpal joints, and it was concluded that MR imaging could successfully be used in the intact joint to detect minor cartilage and bone lesions not visualized by either radiography or macroscopic examination. Hence, MR imaging can be used to delineate interactions between articular cartilage and subchondral bone over time and in vivo.     

Use of laser rhinoscopy to treat a nasal obstruction in a captive California sea lion (Zalophus californianus).

Laser rhinoscopy was used to treat a nasal obstruction in a captive California sea lion (Zalophus californianus). The rehabilitated, adult, female sea lion developed mucopurulent, intermittent, bilateral nasal discharge and functional nasal obstruction 20 mo after acquisition by the Aquarium of the Pacific in Long Beach, California. A 3-mm-thick soft tissue structure spanning the region between the soft and hard palates, a deviated nasal septum, and several nasopharyngeal polyps were identified. Biopsies and cultures of the obstructive web showed ulcerative granulation tissue with suppurative inflammation, bacterial infection, and a partial section of an arthropod larva (not speciated). Laser rhinoscopy was performed to relieve the caudal nasopharyngeal obstruction and ablate the polyps. The sea lion appeared to breathe through the nares with lessened nasal discharge for a period of 6 wk after laser therapy, but within 8 wk the mucopurulent nasal discharge returned, the obstruction had reformed, and the sea lion was euthanized. Postmortem examination confirmed antemortem diagnoses of caudal nasopharyngeal obstruction secondary to inflammatory tissue; however, no additional sections of arthropod parasites were located microscopically.     

San Miguel sea lion virus fed to mink and pigs.

Mink became infected with San Miguel sea lion virus when fed ground meat from seal carcasses showing vesicular-like lesions in the skin. The mink also contracted the infection when they were fed San Miguel sea lion virus infected pig meat or cell culture propagated virus. San Miguel sea lion virus infection in mink was inapparent but the virus was isolated from blood and rectal swabs. Pigs treated similarly with the same virus preparations given to mink developed a severe vesicular disease syndrome similar to that produced by vesicular exanthema of swine. In a separate trial, pigs fed a large sample of commercial ground seal meat did not develop disease signs or antibodies. Further work is needed to assess the hazard of introducing San Miguel sea lion virus into swine on the same premises when potentially San Miguel sea lion virus infective seal meat is fed to mink.     

Epiphyseal cartilage canal blood supply to the metatarsophalangeal joint of foals

Reasons for performing study: It is presently unknown whether cartilage ischaemia plays any part in the pathogenesis of osteochondral fragmentation within the equine metatarsophalangeal joint, as no detailed studies on microcirculation in the area have been reported. Objective: To describe the developmental pattern of the blood supply to the epiphyseal growth cartilage in the metatarso‐phalangeal joint of foals. Methods: Eight Standardbred foals were sacrificed between birth and age 7 weeks to undergo a barium perfusion procedure to demonstrate vessels within growth cartilage canals of one hindlimb. The metatarso‐phalangeal joint was cleared in methyl salicylate and perfused vessels studied in the intact bones. The bones were sawed into 5 mm thick slabs, decalcified and radiographed. Selected slabs were cleared in methyl salicylate for a second time and examined at low magnification. The dorsal half of the sagittal ridge of the distal third metatarsal bone and the plantar half of the proximal phalanx were examined histologically. Results: Regions of the epiphysis with thick cartilage contained a greater number of perfused vessels than regions with thin cartilage. The cartilage canal vessels were oriented either parallel or perpendicular to the underlying ossification front. Cartilage canal vessels were incorporated into the ossification front during growth and became reliant on a subchondral arterial source. Macroscopically visible lesions were not detected in the current group of foals. On histological examination, pathological changes consisting of an area of chondronecrosis surrounded by fibrovascular granulation tissue were found in sections from the lateral proximo‐plantar eminence of the proximal phalanx in the 7‐week‐old foal. Conclusion: The same anatomical feature (traversing the ossification front to enter cartilage canals) reported to render vessels vulnerable to failure in the tarsus was also present in the metatarso‐phalangeal joint of foals. Potential relevance: Osteochondrosis may occur by the same pathogenetic mechanism in the metatarso‐phalangeal joint as in the tarsus of foals.     

MAGNETIC RESONANCE ANATOMY OF THE CARPUS OF THE HORSE DESCRIBED FROM IMAGES ACQUIRED FROM LOW‐FIELD AND HIGH‐FIELD MAGNETS

Cadaver carpi of 30 mature horses with no history of carpal or proximal metacarpal pain were examined using low‐field (0.27 T) and high‐field (1.5 T) magnetic resonance imaging (MRI). Normal MRI anatomy in transverse, sagittal, and dorsal plane images was determined by comparison with anatomical specimens and standard texts. Subchondral bone and cortical bone thickness measurements were obtained from standardised sites. There was variable subchondral bone thickness in the radius and carpal bones; subchondral bone thickness was consistently larger at dorsal compared with palmar sites in the proximal row of carpal bones. The endosteal surface of the subchondral bone was smooth. The shape of the ulnar carpal bone was variable and one or more small osseous fragments were identified palmar to the bone in 5/30 limbs. There was no evidence to suggest that these were pathological fractures or avulsions of the lateral palmar intercarpal ligament. The amount of muscle tissue in the superficial and deep digital flexor tendons in the proximal aspect of the carpus varied, but none was present at the level of the middle carpal joint and distally. Several structures could be evaluated that cannot be imaged using radiography, ultrasonography, or arthroscopy, including the transverse intercarpal ligaments, the radiocarpal ligament, the short palmar carpal ligaments, and the carpometacarpal ligaments. Anatomical variations not previously described were identified, including the layers of the medial aspect of the carpal fascia. Knowledge of the variation in MRI appearance of the carpus of nonlame horses is helpful for interpretation of MR images of lame horses.