Detection and characterization of leptospiral antigens using a biotin/avidin double-antibody sandwich enzyme-linked immunosorbent assay and immunoblot.

A biotin/avidin double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of antigens of Leptospira interrogans serovars in experimentally inoculated bovine urine samples was evaluated. Immunoglobulin G (IgG) from rabbits immunized with L. interrogans serovar hardjo type hardjobovis sonicated, whole cell, and formalinized-heated antigen preparations were purified by a protein A-superose column coupled to fast protein liquid chromatography, and evaluated for species specificity in the ELISA. The ELISA using each specific IgG detected as few as 10(4) leptospires of the homologous serovar hardjo diluted in phosphate-buffered saline solution with Tween 20 (PBSS-Tween 20). On immunoblot analysis of proteinase-K-digested whole cell leptospiral preparations, each IgG revealed the presence of bands specific to serovar hardjo, suggesting the presence of serovar-specific epitopes on the lipopolysaccharide molecules. The minimum number of cells of heterologous serovars pomona, grippotyphosa, bratislava, icterohaemorrhagiae and copenhageni detected by each ELISA was greater, ranging from 10(6) to 10(7). The common antigenic determinants observed on immunoblot analysis were different for each specific IgG, except for a major cross-reacting, possibly flagellar, protein doublet at approximately 36-36.5 kDa. Leptospires were equally well detected by the ELISA in both bovine urine and PBSS-Tween 20.     

Mystery swine disease in The Netherlands: the isolation of Lelystad virus.

In early 1991, the Dutch pig-industry was struck by the so-called mystery swine disease. Large-scale laboratory investigations were undertaken to search for the etiological agent. We focused on isolating viruses and mycoplasmas, and we tested paired sera of affected sows for antibodies against ten known pig viruses. The mycoplasmas M. hyosynoviae, M. hyopneumoniae, and Acholeplasma laidlawii, and the viruses encephalomyocarditis virus and porcine enterovirus types 2 and 7 were isolated from individual pigs. An unknown agent, however, was isolated from 16 of 20 piglets and from 41 of 63 sows. This agent was characterised as a virus and designated Lelystad virus. No relationship between this virus and other viruses has yet been established. Of 165 sows reportedly afflicted by the disease, 123 (75 per cent) seroconverted to Lelystad virus, whereas less than 10 per cent seroconverted to any of the other virus isolates or to the known viral pathogens. Antibodies directed against Lelystad virus were also found in pigs with mystery swine disease in England, Germany, and in the United States. We conclude that infection with Lelystad virus is the likely cause of mystery swine disease.     

Identification by isoelectric focusing of creatine kinase-MM isoforms in the plasma and striated muscle of pigs

Total creatine kinase (CK) and CK-MM isoforms were determined in plasma and longissimus dorsi muscle extracts from normal pigs. Based on their total CK activity, the pigs were divided into two groups. Pigs of group 1 (n=16) had a mean plasma total CK of 298±16 U/L and the distribution of the CK-MM isoforms was 65.7±2.5% CK-MM₃, 18.9±1.6% CK-MM₂ and 15.3±1.5% CK-MM. In group 2 (n=18; 826±75 U/L total CK) four isoforms were observed: 3.1±0.9% CK-MM, 67.9±3.0% CK-MM₃, 21.5±2.3% CK-MM₂ and 7.5±1.3% CK-MM₁. The differences between the two groups of pigs were significant (p<0.001) for CK-MM₁ and the presence of CK-MM. Four CK-MM isoforms were also detected in longissimus dorsi muscle homogenates: 45.6±8.1% CK-MM, 32.6±11.7% CK-MM₃, 16.6±2.3% CK-MM₂ and 5.1±2.8% CK-MM₁. The release of CK-MM isoforms from muscle into plasma seems to be unrelated to the concentration of these isoforms in striated muscle.     

Double-blind evaluation of implants of gold wire at acupuncture points in the dog as a treatment for osteoarthritis induced by hip dysplasia.

Thirty-eight dogs with hip dysplasia were studied to evaluate the use of gold wire implants at acupuncture points around the hip joints. They were assigned at random into two groups of 19. In the treated group, gold wire was inserted through hypodermic needles at electrically found acupuncture points around both hips. In the control group, the areas were prepared in the same way but had only the skin pierced at sites which were not acupuncture points, with a needle of the same size as that used in the treated group. Over a period of six months the dogs were studied repeatedly by two veterinarians and by the dogs' owners who were unaware of the treatments the dogs had received; they assessed the dogs' locomotion, hip function and signs of pain. Radiographs were taken at the beginning and end of the study. Although the data collected from both groups by the veterinarians and the owners showed a significant improvement of locomotion and reduction in signs of pain (P=0.036 for the veterinary evaluation and locomotion and P=0.0001 and P=0.0034 for the owners' evaluation of locomotion and pain, respectively), there were no statistically significant differences between the treated and control groups (P=0.19 and P=0.41, P=0.24, respectively).     

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.     

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.