A recombinant-based feline immunodeficiency virus antibody enzyme-linked immunosorbent assay.

We have developed an antibody detection enzyme-linked immunosorbent assay (ELISA) for the identification of animals infected by feline immunodeficiency virus (FIV). The ELISA solid-phase antigen consists of recombinant FIV gag proteins expressed in bacteria. The proteins are purified from bacterial lysates as insoluble inclusion bodies. In the case of bacterially expressed p24gag, it is shown that all of the linear, sequential epitopes presented by viral p24 during infection are retained. Purified preparations can be substituted for solid-phase whole virus in the IDEXX PetChektm immunoassay. The antibody ELISA duplicates the sensitivity and specificity of the whole virus based PetChek plate assay.     

Use of an indirect enzyme-linked immunosorbent assay (elisa) in the detection of channel catfish, Ictalurus punctatus (Rafinesque), antibodies to Edwardsiella ictaluri

Abstract. The use of an indirect enzyme-linked immunosorbent assay ( elisa ) for the detection of channel catfish antibody to Edwardsiella ictaluri is described. Changes in agglutination titre in fish immunized with Edwardsiella ictaluri heat killed whole bacterins or lipopolysaccharides were reflected by corresponding changes in elisa readings. Relatively high correlations were observed among elisa OD readings, computed elisa titres and corresponding agglutination titres.     

Serum amyloid A protein (SAA) in horses: objective measurement of the acute phase response

A sensitive and precise immunoassay for equine serum amyloid A protein (SAA) was established and used to determine, for the first time, the circulating concentration of this protein in health and disease. As in other species, equine SAA was present only at trace levels in healthy animals but behaved as an extremely sensitive and rapidly responding acute phase reactant following most forms of tissue injury, infection and inflammation, objectively reflecting the extent and activity of disease. Measurements of SAA should make a significant contribution to diagnosis and management of viral and bacterial infection in horses, and routine serial assays could provide an objective criterion for monitoring prospectively the general health of horses in training and racing.     

Incidence of microscopically detectable degenerative characteristics in skeletal muscle of turkey

1. The incidence of microscopically detectable degenerative characteristics in 5 skeletal muscles (m. pectoralis thoracicus, m. supracoracoideus, m. biceps femoris, m. semitendinosus, m. femorotibialis medius) of turkeys was investigated. 2. Samples were obtained from 30 Large White turkey males 14, 16 and 18 weeks old. Hyaline degeneration, infiltration of mononuclear cells and necrotic fibres were observed. 3. Individual fibres varied greatly in size and muscle fibre nuclei were often shrunken and pyknotic. 4. Weak and/or uniform reaction for Ca++-ATPase and SDH in all types of muscle fibres and loss of alkaline phosphatase activity in cell membranes were noted. A positive reaction for acid phosphatase occurred in regions of perivascular infiltration and in necrotic muscle fibres. The majority of muscle fibres possessed high activity for phosphorylase a and b. 5. Based on the use of fluorescein alpha-bungarotoxin conjugate, motor end-plates appeared to be morphologically intact. Direct immunofluorescence with anti-chicken IgG showed positive reaction in muscle fibres undergoing necrosis and in the involved connective tissue. 6. Degenerative changes varied with age and were most marked in the oldest birds. 7. Because gross degenerative symptoms were absent from both the birds and the meat from them, the condition appears to be either different from or a precursor to the degenerative myopathy characterised by other authors.     

Requirement for MHC class II positive accessory cells in an antigen specific bovine T cell response

Purified populations of bovine antigen presenting cells (APCs) and T cells have been isolated from peripheral blood and characterised using various monoclonal antibodies (mAbs) for cell surface markers. Bovine APCs were found in an adherent cell fraction and were non-specific esterase positive, phagocytic and expressed bovine major histocompatibility complex (MHC) class II determinants, all of which are typical macrophage characteristics. T cells were rigorously depleted of accessory cell function before being used in an antigen presenting cell assay. The generation of T helper cells in response to the soluble antigen, ovalbumin, was entirely dependent upon a critical number of APCs. Further the proliferative response was inhibited by several mAbs to bovine MHC class II molecules. Thus the interaction between bovine APCs and helper/inducer T lymphocytes (TH/I) appears to be similar to that in other species.     

The oxygen uptake capacity of swine

Comprehensive studies into domestic pigs and wild boars together with literature data provided a basis for an assessment of aerobic metabolic capacity (VO2max) of swine. The values quoted, from 20 to 94 ml/min-1/kg, had been due to several factors of methodological approach, growth, training, and nutrition as well as to pathophysiological aspects. For full capacity utilisation of VO2max, untrained pigs can be challenged at belt velocities between 0.8 and 1.8 m/s-1 and trained animals at 5 m/s-1.     

Host immune response to northern fowl mite: immunoblot and lectin blot identification of mite antigens

White leghorn hens were experimentally infested with northern fowl mites (Ornithonyssus sylviarum) and antibody responses to mite immunogens were monitored over 12 weeks. Mite burdens increased during the early phase of infestation and declined over the latter weeks of the study. Antigen was prepared from homogenized whole mites, which were then sonicated and extracted with non-ionic detergent. Antigen extract was fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and antibody-reactive polypeptides were identified by immunoblotting. At the start of infestation, hens had natural, pre-existing antibodies that reacted with several mite-extract components. Individual hens had different natural antibody reactivities; however, all birds had immunoglobulins reactive with extract polypeptides of 117,000, 77,000 and 36,000 molecular weight. A variety of mite extract components reacted with hen antibodies generated in response to experimental infestation. The number of antibody-reactive mite polypeptides increased through week 8 of infestation and then decreased by week 12. Fifteen polypeptides of northern fowl mite extract were reactive with antibodies developed by the majority of infested birds. These commonly reactive polypeptides had molecular weights ranging from 40,000 to 160,000. Glycoconjugates of fractionated mite extract were identified by blotting with lectins that have different carbohydrate binding specificities. Also identified were lectins that bound extract components with the same molecular weights as those moieties complexed by immunoglobulins of infested birds.