Immunohistochemical detection of scrapie prion proteins in clinically normal sheep in Pennsylvania.

Following diagnosis of scrapie in a clinically suspect Suffolk sheep, 7 clinically normal flockmates were purchased by the Pennsylvania Department of Agriculture to determine their scrapie status using an immunohistochemical procedure. Two of the 7 euthanized healthy sheep had positive immunohistochemical staining of the prion protein of scrapie (PrP-Sc) in their brains, nictitating membranes, and tonsils. The PrP-Sc was localized in the areas of the brain where, histopathologically, there was neurodegeneration and astrocytosis. The PrP-Sc occurred within germinal centers of the affected nictitating membranes and tonsils and was located in the cytoplasm of the dendrite-like cells, lymphoid cells, and macrophages. These results confirm that immunohistochemical examination of the nictitating membrane can be used as a screen for the presence of scrapie infection in clinically normal sheep at a capable veterinary diagnostic laboratory. In sheep with a PrP-Sc-positive nictitating membrane, the diagnosis of scrapie should be confirmed by histopathology and immunohistochemical examination of the brain following necropsy. Following full validation, immunohistochemistry assays for detection of PrP-Sc in nictitating membrane lymphoid tissues can improve the effectiveness of the scrapie control and eradication program by allowing diagnosis of the disease in sheep before the appearance of clinical signs.     

Humoral immune responses of black-footed penguins (Spheniscus demersus) after DNA-mediated immunization with a beta-galactosidase reporter gene.

Humoral immune responses of black-footed penguins (Spheniscus demersus) to DNA-mediated immunization with a beta-galactosidase reporter gene expression plasmid were evaluated. Six male and 6 female adult penguins received either test plasmid, pCMV-beta, containing the beta-galactosidase gene or control plasmid, pCI, lacking a gene for expression. Three birds from each group were used previously in a diluent control group and given one injection of sterile saline. All samples were screened for anti-beta-galactosidase antibodies by indirect enzyme-linked immunosorbent assay with anti-chicken immunoglobulin G as secondary antibody. Antibodies to beta-galactosidase were detected in the sera of pCMV-beta-inoculated penguins, with a peak response on day 21. Antibody titers of the test plasmid group versus both control groups on days 21, 28, and 42 differed significantly. These results demonstrate that black-footed penguins can be safely transfected with the gene encoding beta-galactosidase and will mount a humoral response against the in vivo-expressed protein. Knowledge from this initial study can be applied to the development of DNA-mediated vaccines against specific infectious diseases of penguins.     

Postovulatory Effect of Intravenous Administration of Lipopolysaccharide (Escherichia coli, O55:B5) on the Endocrine Status of Recently Ovulated Sows

The effects of lipopolysaccharide ( Escherichia coli , O55:B5), administered 18 h after ovulation in the second oestrus after weaning on the hormonal profiles in 14 Swedish cross-bred (Landrace × Yorkshire) multiparous sows were studied. The endotoxin group (E-group) sows were administered with 300 ng/kg of lipopolysaccharide (LPS) whereas the control group (C-group) sows were administered 5 ml of saline intravenously via an indwelling jugular cannula. Blood samples for hormonal analyses were collected from all sows until slaughter. In the E-group, progesterone, cortisol and prostaglandin F_2α metabolite levels increased significantly (p < 0.05) following LPS compared with the C-group. It can be concluded from this study that apart from elevating cortisol and prostaglandin F_2α metabolite, LPS also elevates progesterone levels.     

The Effect of Size and Diet on Gonad Production by the Chilean Sea Urchin Loxechinus albus

Conadal production was studied in Loxechinus albus (Molina, 1782) maintained in cages suspended from a long-line in the Estero Castro, Chiloé for 3 mo during the austral summer. The sea urchins were fed either an artificial diet or a natural diet consisting of the macroalgae Macrocystis pyrifera and Ulva sp. Both diets were tested for four size ranges: 40–45, 50–55, 60–65 and 70–75 mm diameter. For all four size ranges, highest gonad production was with the artificial diet. Gonad production was greatest in the 40–45 mm individuals with an increase of about 1,400% and 750% in the wet weight of gonads in individuals fed the artificial and natural diet, respectively. With the three other size ranges, the increase was nearly 100% with the artificial diet and nearly 0% with the natural diet The gonad index showed similar patterns, being highest in the smallest individuals. Small individuals fed the artificial diet would provide the most cost effective aquaculture as production is best. It is not necessary to grow L. albus to the minimal legal size for fisheries (70-mm diameter) for cost effective gonad production.     

STUDIES ON THE MANAGEMENT OF CARDIOPULMONARY EMBARRASSMENT DUE TO THIOPENTONE ANAESTHESIA IN THE EQUINE

Cardio pulmonary embarrassment was induced with thiopentone sodium in 35 healthy adult donkeys divided in to seven groups of five animals. “To effect” anaesthetic dose was (12.72 ± 0.69 mg/kg) which caused transient hypotension, tachycardia and hypoxemia. A stable fall in MAP 1. 6.77 ± 0.287 Kpa (50.78 ± 2.15 mmHg) and a flat EEG were considered to indicate acute circulatory insufficiency due to thiopentone overdose.     

A Comparison of Arterial and Lingual Venous Blood Gases in Anethetized Dogs

The purpose of this study was to determine whether lingual vanous blood gas samples reflect arterial acid-base gas status in anethetized dogs. Heparinized blood samples were drawn simultaneously from the lingual vein and a peripheral artery in 50 anestheized dogs that were clinical surgical patients, as well as from four experimental dogs in which hemorrahaic shock was being studied. Blood pH, oxygen tension (PO₂), and bicarbonate (HCO_3-)) from the two sources in clinical patients showed significant liner correlation, although arterial PO₂)(PaO₂)) tended to be approximately 110mm Hg higher than lingual venous PO₂). During hemorrahgic shock, however, PaO₂) and PaCO₂) were significantly different from lingual venous PO₂) and PCO₂), Lingula venous blood gas analysis may be useful in assessing acid-base and blood gas status in routline cases, but should not be relied upon in dogs with low cardiac output or poor perfusion.     

Characterization of and radioimmunoassay for canine thyroglobulin

Canine thyroglobulin (cTg) has been isolated and purified. It has similar electrophoretic patterns as Tg from other mammalian species. The main fraction had a MW of 660,000, whereas also fractions of a MW of approximately 1,300,000 (dimer) and 330,000 (subunit) were present. The iodine content was 0.8 to 1.0 % (w/w). cTg did not cross-react with antibodies against human Tg to a degree that would allow the use of a radioimmunoassay for human Tg for the determination of cTg in serum or plasma. Therefore a polyclonal antiserum was raised against cTg and a homologous radioimmunoassay was developed, which was sensitive (0.4 μg/l) and specific (cross-reactivity in cTg assay of human Tg, goat Tg, T₄, T₃, and DIT < 0.01 %).

Plasma Tg levels in normal dogs of both sexes and aged 3–15 years amounted to 192 ± 73 μg/l (mean ± SD, n=30). There was no relation between plasma Tg and T₄ levels.