Lymphocytes of dogs immunised with purified excreted-secreted antigens of Leishmania infantum co-incubated with Leishmania infected macrophages produce IFN gamma resulting in nitric oxide-mediated amastigote apoptosis

The role of nitric oxide (NO) in the anti-leishmanial activity has been confirmed both in vitro and in vivo. Recently, we demonstrated that NO-mediated apoptosis-like amastigote death pathway is an important and highly regulated mechanism used for the clearance of Leishmania within infected murine macrophages stimulated to produce NO endogenously. To further characterize these important effector mechanisms in dog, a natural host-reservoir of L. infantum/L. chagasi, we have developed an ex vivo infection model of canine macrophages. Exposure of L. infantum-infected macrophages to autologous peripheral lymphocytes derived from dogs immunised with purified excreted-secreted antigens of L. infantum promastigotes (LiESAp) formulated with muramyl dipeptide (MDP) as adjuvant resulted in a significant leishmanicidal effect due to interferon (IFN)-gamma dependent macrophage activation. Concomitant accumulation of NO(3)(-)/NO(2)(-) in supernatants of co-cultured cells and in situ staining of parasites with terminal deoxynucleotidyltransferase-mediated dUTP nick-end labelling (TUNEL) and YOPRO-1 showed that NO-mediated apoptosis of intracellular L. infantum amastigotes is occurring in canine macrophages as previously observed in mouse models. Monitoring these parameters in dogs after immunisation and before experimental challenge can represent a useful and easy way to rapidly evaluate vaccine candidates against canine visceral leishmaniasis.     

Enhancement of bone healing using non-glycosylated rhBMP-2 released from a fibrin matrix in dogs and cats

OBJECTIVES: To test a non-glycosylated recombinant human bone morphogenetic protein-2 (ngly-rhBMP-2)/fibrin composite, which has been shown experimentally to enhance healing of bone defects in rodents, in a clinical case series of dogs and cats undergoing treatment for fracture non-unions and arthrodesis. METHODS: A ngly-rhBMP-2/fibrin composite was applied in 41 sites in 38 dogs and cats for which a cancellous bone autograft was indicated, replacing the graft. RESULTS: Bridging of the bone defect with functional bone healing was achieved in 90 per cent of the arthrodesis and fracture nonunions treated in this manner. CLINICAL SIGNIFICANCE: This prospective clinical study demonstrates the beneficial effects of ngly-rhBMP-2 in a specially designed fibrin matrix on the treatment of bone defects, and validates the use of this composite as an alternative to bone autografts in dogs and cats.     

Treatment of 10 dogs with discospondylitis by fluoroscopy-guided percutaneous discectomy

Ten dogs suffering from discospondylitis were treated by percutaneous discectomy and local and systemic antibiotic therapy. With fluoroscopic guidance, a cylinder 5 mm in diameter was removed from the centre of the intervertebral space, yielding a fenestration and decompressing the disc without producing any spinal instability. The causative bacteria were identified in nine of the 10 biopsy specimens, but in only three urine cultures and four blood cultures. In two cases, the antibiotics used initially had to be changed owing to the organisms' antibiotic resistance. The clinical signs of the dogs improved markedly after two to nine days (mean 4.2 days) and had resolved completely after five to 14 days (mean 9.1 days). In all the cases the disease could be classified histologically as either acute or chronic, and the disease was classified as chronic in one dog. No side effects were observed.     

Effects of dietary cation-anion difference on blood cortisol and ACTH levels in reproducing ewes

Formulation of rations to induce a compensated metabolic acidosis in the post-partum cow has proved a useful strategy for prevention of milk fever. Such acidification improves the ability of the animal to maintain calcium homeostasis by promoting the absorption of calcium from the intestine and mobilization of calcium from the bone. In humans, an acidogenic diet results in mild metabolic acidosis in association with a state of cortisol excess and this increase in plasma cortisol may increase bone catabolism. The objective of our experiment was to induce acidification by anionic salt supplementation and to study the effects of anionic supplementation on plasma cortisol and adrenocorticotropic hormone (ACTH) levels in sheep. Twenty-seven twin-bearing sheep were assigned to two experimental groups and a control group, depending on dietary cation-anion difference (DCAD) (+272.6, -88.9 and +164.5 mEq/kg DM respectively). Sheep assigned to each dietary treatment received their respective rations beginning 6 weeks pre-partum and continuing until 12 days post-partum. Anionic diet induced a non-respiratory systemic acidosis in association with a mild increase in plasma cortisol concentration without changes in plasma ACTH levels. Our data suggest that the mild hypercortisolism observed in sheep fed the anionic diet may not be an effector for bone resorption induced by anionic salts. A mild hypercortisolism of this magnitude may lead to osteoporosis but this might require many years of adrenal hypersecretion while anionic salts are only used during the last weeks of pregnancy.     

Neurogenesis in a young dog with epileptic seizures

Epileptic seizures can lead to various reactions in the brain, ranging from neuronal necrosis and glial cell activation to focal structural disorganization. Furthermore, increased hippocampal neurogenesis has been documented in rodent models of acute convulsions. This is a report of hippocampal neurogenesis in a dog with spontaneous epileptic seizures. A 16-week-old epileptic German Shepherd Dog had marked neuronal cell proliferation (up to 5 mitotic figures per high-power field and increased immunohistochemical expression of proliferative cell nuclear antigen) in the dentate gyrus accompanied by microglial and astroglial activation. Some granule cells expressed doublecortin, a marker of immature neurons; mitotically active cells expressed neuronal nuclear antigen. No mitotic figures were found in the brain of age-matched control dogs. Whether increased neurogenesis represents a general reaction pattern of young epileptic dogs should be investigated.     

Echocardiographic parameters in healthy young adult Sphynx cats

The objective of this retrospective study is to determine normal reference values for 2-Dimension (2D) and Motion-mode (M-mode) echocardiographic parameters in nonsedated healthy young adult Sphynx cats and to compare them to those of the domestic shorthair (DSH). 131 Sphynx cats underwent cardiac screening prior to breeding. The control group consisted of 30 healthy adult domestic cats. A complete cardiac ultrasound was performed on all cats using right parasternal long and short axis views. There were few echocardiographic parameters in the Sphynx that differed from those of the healthy DSH. Only the left atrial (LA) dimension in 2D and M-mode, the left atrial/aortic (LA/Ao) ratio and the internal dimension of the left ventricle in systole (LVIDs) measured with M-mode were different. In conclusion, although the heart of Sphynx cat can often have a particular 2-D echocardiographic appearance, the M-mode cardiac dimensions are similar to those of the DSH.     

Response of white leghorn chickens to infection with avian leukosis virus subgroup J and infectious bursal disease virus

The effects of viral-induced immunosuppression on the infectious status (viremia and antibody) and shedding of avian leukosis virus (ALV) were studied. Experimental white leghorn chickens were inoculated with ALV subgroup J (ALV-J) and infectious bursal disease virus (IBDV) at day of hatch with the ALV-J ADOL prototype strain Hcl, the Lukert strain of IBDV, or both. Appropriate groups were exposed a second time with the Lukert strain at 2 wk of age. Serum samples were collected at 2 and 4 wk of age for IBDV antibody detection. Samples for ALV-J viremia, antibody detection, and cloacal shedding were collected at 4, 10, 18, and 30 wk of age. The experiment was terminated at 30 wk of age, and birds were necropsied and examined grossly for tumor development. Neoplasias detected included hemangiomas, bile duct carcinoma, and anaplastic sarcoma of the nerve. Control birds and IBDV-infected birds were negative for ALV-J-induced viremia, antibodies, and cloacal shedding throughout experiment. By 10 wk, ALV-J-infected groups began to develop antibodies to ALV-J. However, at 18 wk the incidence of virus isolation increased in both groups, with a simultaneous decrease in antibody levels. At 30 wk, 97% of birds in the ALV-J group were virus positive and 41% were antibody positive. In the ALV-J/IDBV group, 96% of the birds were virus positive at 30 wk, and 27% had antibodies to ALV-J. In this study, infection with a mild classic strain of IBDV did not influence ALV-J infection or antibody production.     

Disseminated pheochromocytoma in a North American river otter (Lontra canadensis)

A 21-yr-old male North American river otter (Lontra canadensis) with a chronic history of degenerative osteoarthritis was evaluated for acute posterior paralysis. Because no definitive cause was identified and a poor prognosis was expected, the otter was euthanatized. A malignant neoplasm of adrenal gland origin with disseminated metastases to the central nervous system, lymph nodes, diaphragm, pancreas, spleen, and liver was diagnosed on postmortem examination. No clinical signs of disseminated neoplasia had been noted throughout the otter's history. The adrenal neoplasm was composed of nests of epithelial cells surrounded by a fine fibrovascular stroma. Neoplastic cells were immunohistochemically positive for chromogranin A, PGP9.5, metencephalin, and endorphin and negative for melan A and inhibin, confirming a diagnosis of a malignant pheochromocytoma. On the basis of the necropsy finding, metastasis of the pheochromocytoma might have contributed to the observed clinical signs.