Maternal HMB treatment affects bone and hyaline cartilage development in their weaned piglets via the leptin/osteoprotegerin system

It has been demonstrated in animal studies that prenatal administration of β‐hydroxy‐β‐methylbutyrate (HMB, metabolite of leucine) influences general growth and mechanical endurance of long bones in newborn offspring in sex‐dependent manner. The present experiment was conducted to evaluate the effect of HMB treatment of pregnant sows on bone development in offspring at weaning. From 70th day until the 90th day of gestation, sows received either a basal diet (n = 12) or the same diet supplemented with HMB (n = 12) at the dose of 0.2 g/kg of body weight/day. Femora obtained from six males and females in each group weaned at the age of 35 days were examined. Maternal HMB treatment significantly enhanced body weight and changed bone morphology increasing femur mechanical strength in both sexes. Maternal HMB supplementation also elevated bone micro‐ and macroelement concentrations and enhanced content of proteoglycans in articular cartilage. Based on the obtained results, it can be concluded that maternal HMB supplementation in the mid‐gestation period significantly accelerated bone development in both sexes by upregulation of a multifactorial system including leptin and osteoprotegerin. However, the sex (irrespective of the HMB treatment) was the factor which influenced the collagen structure in cartilages and trabecular bone, as demonstrated both by the Picrosirius red staining and performed analysis of thermal stability of collagenous tissues. The structural differences in collagen between males and females were presumably related to a different collagen maturity. No studies conducted so far provided a detailed morphological analysis of bone, articular cartilage, growth plate and the activities of the somatotropic and pituitary–gonadal axes, as well as leptin/osteoprotegerin system in weaned offspring prenatally treated with HMB. This study showed also the relationship between the maternal HMB treatment and bone osteometric and mechanical traits, hormones, and growth and bone turnover markers such as leptin, osteoprotegerin and insulin‐like growth factor‐1.     

Biochemical characterisation of navicular hyaline cartilage,navicular fibrocartilage and the deep digital flexor tendon in horses with navicular disease

The study hypothesis was that navicular disease is a process analogous to degenerative joint disease, which leads to changes in navicular fibrocartilage and in deep digital flexor tendon (DDFT) matrix composition and that the process extends to the adjacent distal interphalangeal joint. The objectives were to compare the biochemical composition of the navicular articular and palmar cartilages from 18 horses with navicular disease with 49 horses with no history of front limb lameness, and to compare navicular fibrocartilage with medial meniscus of the stifle and collateral cartilage of the hoof. Cartilage oligomeric matrix protein (COMP), deoxyribonucleic acid (DNA), total glycosaminoglycan (GAG), metalloproteinases MMP-2 and MMP-9 and water content in tissues were measured. Hyaline cartilage had the highest content of COMP and COMP content in hyaline cartilage and tendon was higher in lame horses than in sound horses (p<0.05). The concentration of MMP-2 amount in hyaline cartilage was higher in lame horses than in sound horses. The MMP-2 amounts were significantly higher in tendons compared to other tissue types. Overall, 79% of the lame horses with lesions had MMP-9 in their tendons and the amount was higher than in sound horses (p<0.05). In horses with navicular disease there were matrix changes in navicular hyaline and fibrocartilage as well as the DDFT with potential implications for the pathogenesis and management of the condition.     

Influence of interleukin-1beta and hyaluronan on proteoglycan release from equine navicular hyaline cartilage and fibrocartilage

Proteoglycan (PG) release, in response to recombinant human interleukin-1beta (rh-IL-1beta), was measured in cartilage explants obtained from the equine distal sesamoid bone (navicular bone). Fibrocartilage from the surface of the navicular bone apposing the deep digital flexor tendon and hyaline cartilage from the surface of the navicular bone articulating with the middle phalanx were labelled with 35SO4. Hyaline cartilage from the distal metacarpus was used as a control tissue. Following radiolabel incorporation, the three cartilage types were treated with rh-IL-1beta (100 U/mL) in the presence of hyaluronan (0.2, 2, 20, 200 and 2000 microgram/mL). rh-IL-1beta-Induced PG release was measured by scintillation assay of PG-bound radiolabel. Increases in PG release of 94% (P < 0.01), 101% (P < 0.05) and 122% (P < 0.05), in response to rh-IL-1beta, were noted in fibrocartilage, navicular hyaline cartilage and metacarpal hyaline cartilage, respectively. Hyaluronan (0.2 microgram/mL) significantly reduced rh-IL-1beta-induced PG release in metacarpal hyaline cartilage (P < 0.01). In fibrocartilage and navicular hyaline cartilage, hyaluronan did not reduce PG release and at some concentrations appeared to increase PG release, although this was not statistically significant. These experiments show that (i) fibrocartilage and hyaline cartilage of the navicular bone release PGs in response to rh-IL-1beta, and (ii) hyaluronan does not prevent rh-IL-1beta-induced breakdown of navicular bone cartilage.     

Analysis of cartilage oligomeric matrix protein and matrix metalloproteinase-9 in cerebrospinal fluid of miniature dachshund with intervertebral disc herniation

We evaluated whether the cerebrospinal fluid (CSF) concentration of cartilage oligomeric matrix protein (COMP) is related to disease severity, prognosis and matrix metalloproteinase (MMP)-9 activity of the CSF in miniature dachshund with intervertebral disc herniation. Samples were obtained from 23 patients and 6 normal dogs, and all patients received hemilaminectomy. Twenty dogs recovered successfully and 3 of 11 dogs without deep nociception had MMP-9 activity in the CSF and an unsuccessful outcome. The COMP levels from patients were significantly higher than those from normal dogs. MMP-9 activity and neurological severity were not related to the COMP levels. However, the COMP levels from 3 unsuccessful cases that had MMP-9 activity were significantly lower than those from all recovered cases and/or successful cases without deep nociception. Concerning severe cases, increased proteolytic activity might affect the COMP concentration and prognosis due to MMP-9 associated deleterious effects.     

Analysis of cartilage oligomeric matrix protein and matrix metalloproteinase-9 in cerebrospinal fluid of miniature dachshund with intervertebral disc herniation

We evaluated whether the cerebrospinal fluid (CSF) concentration of cartilage oligomeric matrix protein (COMP) is related to disease severity, prognosis and matrix metalloproteinase (MMP)-9 activity of the CSF in miniature dachshund with intervertebral disc herniation. Samples were obtained from 23 patients and 6 normal dogs, and all patients received hemilaminectomy. Twenty dogs recovered successfully and 3 of 11 dogs without deep nociception had MMP-9 activity in the CSF and an unsuccessful outcome. The COMP levels from patients were significantly higher than those from normal dogs. MMP-9 activity and neurological severity were not related to the COMP levels. However, the COMP levels from 3 unsuccessful cases that had MMP-9 activity were significantly lower than those from all recovered cases and/or successful cases without deep nociception. Concerning severe cases, increased proteolytic activity might affect the COMP concentration and prognosis due to MMP-9 associated deleterious effects.     

Intracytoplasmic hyaline globules in the adrenal medulla of laboratory animals.

Intracytoplasmic hyaline globules were found frequently in chromaffin cells of the adrenal medulla from several species of laboratory animals that died from various causes. The globules stained intensely red with PAS, were weakly acid-fast, did not stain with oil-red O and showed bright yellow autofluorescence. Ultrastructurally, they were finely granular, round to oval, intensely osmiophilic and were surrounded by an intimately bound trilaminar membrane. The globules were interpreted as aggregated secretory granules that occur in degenerating cells.     

Rhabdomyosarcoma of the tongue in a dog

A lingual rhabdomyosarcoma was diagnosed in a six-year-old Staffordshire bull terrier with clinical signs of dysphagia. The unsuccessful response to initial chemotherapy, to surgical resection and subsequent surgical resection and radiation therapy is documented. The accurate diagnosis and prognosis of such tumours is also discussed. Lingual rhabdomarcoma has not previously been reported in the dog.     

Idiopathic hyperchylomicronaemia in miniature schnauzers

Several authors have reported the presence of fasting hyperlipidaemia in adult miniature schnauzers presented with either paroxysmal abdominal pain, vomiting or diarrhoea. Acute pancreatitis is commonly confirmed. There appears to be no sex predilection for the condition although the majority of affected dogs are middle aged and older. Lipid profiles of affected dogs reveal extreme elevations of fasting serum triglyceride (TG) concentrations ranging from 5·5 mmol/litre to over 90 mmol/litre; the hyperlipidaemia is almost exclusively attributed to chylomicron excess. To date, the reviewer's studies have not shown a correlation between TG levels and severity of signs. However, miniature schnauzers with fasting TG levels greater than 5·5 mmol/litre are considered to be at significant risk and should be further evaluated as candidates for dietary intervention.     

Besnoitiosis in a miniature donkey

A 1-year-old male miniature donkey ( Equus asinus ) from a herd of eight was presented with a 9-month history of pruritic dermatitis, lethargy and anorexia. Physical examination revealed diffuse lichenification and scales involving the skin of the face, head and dorsum from the neck to the pelvis. The main histological alteration within the superficial and deep dermis was the presence of multiple large, spherical, thick walled, protozoal Besnoitia cysts. In addition, the inflammatory response consisted of a moderate, superficial and deep perivascular, mixed mononuclear cell infiltrate, with epidermal hyperplasia and compact orthokeratosis. Based upon the large size of the protozoal cysts and the ultrastructural features of the bradyzoites contained therein (conoid, polar ring, rhoptries, micronemes and microtubules), a diagnosis of cutaneous besnoitiosis was established. Treatment with trimethoprim-sulphamethoxazole resulted in significant clinical improvement. To our knowledge, this is the first reported case of besnoitiosis in a miniature donkey in North America.     

Thrombosis of the portal vein in a miniature schnauzer

A miniature schnauzer with a history of apathy, anorexia and jaundice was presented to the Utrecht University Clinic for Companion Animals. Abnormal laboratory findings included highly increased levels of total bile acids and alkaline phosphatase, and hyponatraemia. Abdominal ultrasonography revealed that the right side of the liver was enlarged and the left side was small, together with a thrombus in the portal vein. Biopsies from the right side of the liver demonstrated subacute to chronic active hepatitis, for which the dog was treated with prednisolone (1 mg/kg/day for four weeks). No improvement was observed and the owner requested euthanasia. At necropsy the left lobes of the liver were found to be small and firm, while the right lobes were large and soft. There were two thrombi in the portal vein. Microscopic examination revealed chronic active hepatitis and cirrhosis.     

Hematologic values of the yucatan miniature pig

Mean values and ranges for 16 hematological parameters for healthy, young, sexually-mature Yucatan miniature swine are presented. No significant differences were observed between sexes with regard to hemograms. Comparison of observed values with those published in the literature for standard domestic and other breeds of miniature swine reveals no major differences.     

Ultrastructure of normal epiphyseal cartilage of the articular-epiphyseal cartilage complex in growing swine

Normal epiphyseal cartilage from the articular-epiphyseal cartilage complex (A-E complex) of the distal parts of the femur and humerus of growing commercial crossbred boars was collected, embedded in plastic, and studied by light and electron microscopies. The morphology of this cartilage was determined to provide a basis for comparison with cartilage affected with osteochondrosis, an important clinical disease in swine. Normal epiphyseal cartilage from the A-E complex in growing swine was divided into 4 major regions of cells: resting, proliferating, hypertrophic, and calcifying regions. Cells in the resting zone contained prominent lipid and densely aggregated glycogen. As the cells proliferated and matured, the lipid and glycogen became less prominent. The lipid droplets became smaller and scarcer, and the glycogen became dispersed in the cytoplasm. Proliferating and hypertrophic cells clustered in roughly egg-shaped groups of 4 to 8 cells/plane of section. In the calcifying region, the interterritorial matrix (between cell clusters) calcified, and the territorial matrix (uniting cells in a cluster) remained uncalcified. Calcified matrix extended the depth of one cell group from the area of capillary penetration, and the capillaries invaded by entering a cluster of cells. Territorial matrices in all regions of A-E complex epiphyseal cartilage were composed of randomly oriented collagen fibrils with a granular fibrillar proteoglycan network dispersed between the fibrils. Heterogeneity of chondrocytes was characterized by the presence of both light- and dark-staining cells in the proliferating through calcifying regions and by 3 morphologically distinct light cell types in the late hypertrophic and calcifying regions.