Objective: This research is designed to explore the methods of isolation and culture for endothelial progenitor cells from bovine bone marrow, characteristic, induced differentiative capacity in vitro. Material and methods: Main experimental reagents contain DMEM/F12, fetal bovine serum, percoll lymphocyte separating, Trypsin 1: 250, VEGF, bFGF, GF-1, EDTA and so on. Cultivation system is DMEM/F12 with 10% FBS and VEGF 10 ng/mL, cultured under 37°C, 5% CO2, saturated humidity. Cell viability is measured by trypan blue solution exclusion test. Immunofluorescent detection is used to detected cell surface markers and double swallows, while bovine chromosome is analyzed by karyotyping. Results: We find that the majority of bovine endothelial progenitor cells (EPCs) are fibrous shaped. Frozen survival of bovine EPCs before and after cryopreservation is 95.2±0.14% and 80.9±0.30% respectively; cryopreservation affects little on the viability of bovine EPCs. Immunofluorescent detection of the cell surface markers CD34, CD133 and flk present positive, which can confirm that the cell cultured in vitro are EPCs. Then Dil-ac-LDL and FITC-UAE-1 uptake assays are carried out. Eventually, bovine EPCs are induced to differentiate into endothelial cells and smooth muscle cells respectively, demonstrating the multi-lineage differentiation potential of bovine EPCs in vitro.Conclusion: EPCs can be got with proper culture system. The little cell cryopreservation effect and stronger induced differentiation potential in vitro imply that EPCs can be applied in genetic resources conservation and reuse. [J Adv Vet Anim Res 2017; 4(1.000): 88-96]

This report describes atypical chronic trypanosomosis in a three year male Spitz dog. Fever, lethargy and anorexia were the early presenting signs without any hemato-biochemical abnormality. Peripheral blood smear examination was non-diagnostic on three consecutive times. Trypanosma evansi was confirmed in the Leishman stained thin blood smears (moderate parasetemia) on fourth parasitological examination. Biochemical profile showed a remarkable elevation in total serum bilirubin (6.7 mg%) and activities of alanine amino transferase (ALT) (950 IU/L) and alkaline phosphatase (AKP) (1050 IU/L) after a month. Anemia, leucopenia, neutropenia, lymphopenia and thrombocytopenia suggestive of bone marrow depression appeared by about 73 days of presentation of case. A rapid complete clinical recovery occurred within a week after treatment with quinapiramine sulphate and chloride combination @ 3.5mg/kg bwt. Hemoglobin, leucocyte and thrombocyte count improved within six days, however, liver enzyme activity normalized slowly over three months.

The objective of this study was to determine the feasibility of trephination of the frontal sinus and injection of antifungal cream using a medical bone marrow drill in dogs. Results were compared with frontal sinus trephination using a standard surgical technique. Bilateral trephination of the frontal sinuses was carried out in the heads of 11 cadavers using a medical bone marrow drill and a surgical bone chuck. The time taken to carry out the procedure using both techniques was compared. Before and after injection of antifungal cream into the frontal sinuses, computed-tomography (CT) scanning was done to assess for iatrogenic trauma and to determine the degree to which the sinuses were filled with each technique and evaluate the diffusion of the cream into the nasal cavity of each dog. The mean volume of the sinuses was 8.8 mL (3.1 to 14.3 mL). Trephination, flushing, and injecting of antifungal cream were significantly faster using the medical technique. There was no significant difference in the mean filling of the frontal sinuses between the medical (82.7%) and the surgical (82.4%) technique (P-value = 0.3). Filling of the nasal cavity was classified as very good in 6/11 cases, with evidence of trauma caused by the surgical trephination technique in 1 head. Findings suggest that use of the medical bone marrow drill is highly feasible for frontal sinus trephination. Injection of antifungal cream into the frontal sinuses using the bone marrow needle resulted in good diffusion into the ipsilateral nasal cavity and could be used to treat aspergillosis when debridement or sinusoscopy is not deemed necessary.