Nuclear transfer using a bovine mammary epithelial cell line (BMEC)

The developmental potential of nuclei from a bovine mammary epithelial cell line (BMEC) in nuclear transfer was investigated. For nuclear transfer donors, BMEC cells (passage 15) were cultured for 4–5 days after seeding at cell densities of 1.0 × 10⁵ cells/cm² (high‐density group) or 0.8 × 10⁴ cells/cm² (low‐density group). First, the effective electric stimulation for fusion of enucleated oocytes with BMEC cells was examined. Fusion rates reached maximum with two DC pulses of 30 V/150 µm for 20 µs in the high‐density group and with two DC pulses of 25 V/150 µm for 10 µs in the low‐density group. The fusion rate (37.5%) in the high‐density group was significantly (P < 0.005) lower than in the low‐density group (71.4%). Second, the in vitro developmental potential of nuclear transfer embryos derived from BMEC cells was examined. In the high‐density and low‐density groups, 18.8% and 24.1% of fused oocytes, respectively, developed to blastocyst stage. The results of this study indicate that nuclei from BMEC cells support the development of nuclear transfer embryos to the blastocyst stage and that the efficiency of oocyte–cell fusion is affected by the culture conditions of the donor BEMC cells before nuclear transfer.     

Electron microscopic observation of the respiratory tract of SPF piglets inoculated with Mycoplasma hyopneumoniae.

Seven hysterectomy derived piglets were repeatedly challenged with Mycoplasma hyoneumoniae during the first week of life. Samples of trachea, bronchi and lung tissue collected 2–11 weeks post-inoculation (p.i.) were examined using light and electron microscopy. Autoradiography was used to study in more detail the site of M. hyopneumoniae multiplication. Gross lesions were observed in lung tissue and were characterized by hyperplasia of the epithelium and an increased mononuclear cell accumulation in perivascular and peribronchiolar areas. Mild lesions of the trachea and the bronchi, including epithelial hyperplasia and infiltration of the lamina propria by inflammatory cells, were noted.

Electron microscopy showed that, 2–6 weeks p.i., changes in the mid-trachea and bronchi surface consisted of the loss of cilia. Mycoplasmas covered tufts of cilia remaining on the epithelial cell surface. Scanning and transmission electron micrographs showed that they were predominantly found closely associated with the top of cilia. No specialized terminal structure could be seen and no mycoplasma cells were identified lying free in the lumen nor in close contact with the plasma membrane of cells of microvilli. Some fine fibrils radiating from one mycoplasma to another or to cilia were seen at higher magnification by scanning electron microscopy. Six to eleven weeks p.i., a disrupted epithelial surface lacking cilia was observed. Cells were desquamated and shed into the lumen with cellular remains containing droplets of mucus.

Autoradiography revealed that label corresponded to the observed mycoplasma distribution. At the top of cilia, a high density of labeling was visible in the zone of high mycoplasma concentration. Therefore, incorporation of the label in the mycoplasma is proof or their multiplication in the trachea.

The intimate association between the mycoplasma and cilia may be an important factor in the pathogenesis of the disease caused by M. hyopneumoniae (swine enzootic pneumonia).     

The Effect of Pasteurella haemolytica A2 Infection on Phagocytosis Efficiency of Caprine Broncho‐Alveolar Macrophages

An experiment was designed to study the in vivo effect of Pasteurella haemolytica A2 infection on the phagocytosis activity of caprine broncho‐alveolar macrophages and the extent of pneumonic lesions. Twelve healthy local Kacang goats, about 7 months of age, were divided into two groups of six. Goats in group 1 were inoculated intratracheally with 4 ml inoculum containing 2.8 × 10⁹ colony‐forming units (CFU)/ml of Staphylococcus aureus. Goats in group 2 were inoculated intratracheally with 4 ml of inoculum containing 9.5 × 10⁸ CFU/ml of Pasteurella haemolytica A2 isolated earlier from pneumonic lungs of goat. At intervals of 3 and 7 days post‐challenge five goats from each group were killed and the lungs were washed with sterile phosphate‐buffered saline. Smears were prepared from the lung washing fluid and the number of macrophages with phagocytic activity was determined. At day 3 post‐infection, goats of both groups showed a similar pattern of pneumonic lesion. The lung washing fluid of goats in group 2 was found to contain numerous neutrophils and macrophages. Goats in group 2 showed significantly (P < 0.05) higher extent of lung lesions than group 1. Similarly, the average extent of lung lesions was significantly (P < 0.05) more severe in group 2 at day 7 post‐infection. The lung washing fluid contained mostly macrophages. The phagocytic activity following S. aureus infection was more efficient and significantly (P < 0.01) higher compared with infection by P. haemolytica A2. There were weak correlations between the extent of pneumonic lesion and the phagocytic activity. Thus, goats with poor phagocytic activity were likely to develop more extensive lung lesions.     

Identification of an Intrinsic Platelet Function Defect in Spitz Dogs

A recently identified intrinsic platelet function defect in 2 Spitz dogs is described. Both affected dogs had a history of chronic intermittent bleeding primarily from the nasal, oral, and gastrointestinal mucosa. Platelet aggregation in response to adenosine diphosphate (ADP), collagen, and platelet activating factor (PAF) was absent; however, platelet shape change did occur. Platelets aggregated in response to gamma thrombin, although a delayed onset and a reduced velocity of aggregation were present. Platelet ¹⁴C-serotonin release was diminished in response to collagen and PAF. Glycoprotein Illa was detected on the surface of platelets by flow cytometry. Platelets were morphologically normal under light and electron microscopy. Two male Spitz dogs, related to one of the affected dogs, did not have a bleeding diathesis. Collagen-induced platelet aggregation, however, was diminished in these 2 dogs. This platelet defect most closely resembles the defect described in Basset hounds.     

The effect of copper sulphate on performance and the structure of the gizzard in broilers

Addition of copper sulphate (about 200 mg per kg) to conventional diets produced a small, though variable, growth response and improved food conversion in broilers maintained under commercial conditions. In some experiments interactions were found between sex or coccidiostat and copper in the diet. Broilers fed on diets containing greater concentrations of copper (between 400 and 600 mg per kg) exhibited reduced growth rate and food intake. The gizzard linings of birds receiving diets containing 600 mg CuSO₄ per kg showed extensive damage. This damage was attributable to a massive increase in the shedding of the gizzard glandular cells into the koilin layer and to disruption and cessation of koilin production. Similar though less severe gizzard changes occurred in birds which received 400 mg per kg and there was a slight increase in cellular desquamation in some of the birds receiving 200 mg per kg. High concentrations of copper were found in the gizzard lining and caecal contents of birds receiving supplementary copper and there was a progressive increase in the concentration of copper in the litter on which such birds had been reared.     

Land Cover Type and Fire in Portugal: Do Fires Burn Land Cover Selectively?

The purpose of this study is to investigate if, or under what conditions, fires select given land cover types for burning. If fires burn unselectively then the land cover composition (the proportional area of various land cover types) of individual fires should approximate the land cover composition available in their neighborhood. In this study we test this hypothesis by performing statistical analyses of a data set consisting of paired vectors with the proportions of land cover types present in burned areas and in their respective surroundings. The statistical methods employed (a permutation technique and the C_max statistic) are commonly used in resource selection studies where data is subject to a unit-sum constraint. The results of the analysis of 506 fires that burned in Portugal in 1991 indicate that fires are selective, with small fires exhibiting stronger land cover preferences than large fires. According to the results of a multiple comparison analysis performed for small fires, there is a marked preference for shrubland followed by other forest cover types, while agriculture is clearly avoided. A similar analysis is performed to test if fire selectivity is related to the ecological region where it occurs. The results obtained in this study contribute to the discussion on the relative importance of fuels as a drivers of fire spread.     

Congenital aganglionosis in a 3-day-old Holstein calf

Necropsy of a 3-day-old Holstein heifer revealed proximal megacolon and distal colorectal hypoplasia. Histologically, the hypoplastic distal colon and rectum lacked submucosal and myenteric ganglia. Clinical history, physical examination, and pathologic findings were consistent with intestinal aganglionosis, a congenital anomaly well documented in humans and foals but not previously reported in cattle.