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Serodiagnosis of ovine paratuberculosis, using lipoarabinomannan in an enzyme-linked immunosorbent assay
1989
Sugden, E.A. | Corner, A.H. | Samagh, B.S. | Brooks, B.W. | Turcotte, C. | Nielsen, K.H. | Stewart, R.B. | Duncan, J.R.
The use of lipoarabinomannan (LAM; obtained from Mycobacterium paratuberculosis) in an ELISA (LAM-ELISA) to test 75 sheep sera from a paratuberculosis-infected flock resulted in an approximate threefold increase in sensitivity (from 23.5% to 70.6%), compared with the use of Annau's polysaccharide in a complement fixation test (P-CFT). Even after manipulation of the LAM-ELISA cut-off value to produce a specificity of 100% to match that of the P-CFT, the sensitivity still was approximately twofold greater than that of the P-CFT. Anti-bovine monoclonal antiglobulin-enzyme conjugates matched commercially available anti-ovine polyclonal antiglobulin-enzyme conjugates with respect to sensitivity and specificity. False-positive results were found to be less frequent after combining 2 serodiagnostic tests, LAM-ELISA and D antigenagar gel immunodiffusion, resulting in an increase in specificity from 88.1% to 95.2%. The repeatability of true seropositive and seronegative results was found to be 89.5% and 91.1%, respectively, for sera obtained less than or equal to 1 month prior to slaughter and 91.7% and 95.5%, respectively, for reanalysis of sera obtained at the time of slaughter.
Show more [+] Less [-]Differences in virulence between two strains of Streptococcus suis type II after experimentally induced infection of newborn germ-free pigs
1989
Vecht, U. | Arends, J.P. | Molen, E.J. van der | Leengoed, L.A.M.G. van
Fifteen newborn germ-free pigs were inoculated with 2 strains, D-282 and T-15, of Streptococcus suis type II. Some pigs also were preinoculated with Bordetella bronchiseptica, which successfully predisposed them to S suis infection. The 2 streptococcal strains were differentiated by muramidase treatment, which released certain high molecular-weight proteins, termed muramidase-released proteins (MRP), from the cell wall of strain D-282, but not from the cell wall of strain T-15. Only strain D-282 (MRP-positive) induced clinical signs of disease and markedly increased neutrophil numbers in pigs. Streptococci were more frequently isolated from fecal swab specimens obtained from pigs inoculated with strain D-282 (MRP-positive) than from specimens obtained from pigs inoculated with strain T-15 (MRP-negative). Both strains were isolated from nasal swab specimens obtained from all infected pigs. Postmortem examination revealed fibrinopurulent meningitis, polyserositis, and polyarthritis in pigs inoculated with strain D-282; this strain was isolated from the CNS, serosae, visceral organs, heart, and joints. Whereas strains D-282 caused several pathologic changes, strain T-15, isolated from the lungs, caused only pneumonia. Both strains were isolated from the tonsils of all pigs. Virulence differed distinctly between the MRP-positive and the MRP-negative strains.
Show more [+] Less [-]Antibodies to bovine serum albumin in swine sera: implications for false-positive reactions in the serodiagnosis of African swine fever
1989
Escribano, J.M. | Pastor, M.J. | Sanchez-Vizcaino, J.M.
Antibodies to bovine serum albumin were detected in swine sera by use of an immunoblotting technique. Such sera had false-positive reactions, as determined by results of African swine fever virus serodiagnostic techniques when bovine serum albumin was a contaminant in the soluble cytoplasmic antigen obtained from infected cells cultured in the presence of bovine serum. The soluble cytoplasmic antigen obtained from cell cultures infected with African swine fever virus in the presence of porcine serum did not react with the false-positive sera and, therefore, was used for African swine fever virus serodiagnostic methods, with 0% false-positive results.
Show more [+] Less [-]Variations of serum vitamin E, cholesterol, and total serum lipid concentrations in horses during a 72-hour period
1989
Craig, A.M. | Blythe, L.L. | Lassen, E.D. | Rowe, K.E. | Barrington, R. | Slizeski, M.
Fluctuations of serum vitamin E (alpha-tocopherol), cholesterol, and total lipids were monitored in 12 horses at 3-hour intervals for 72 hours. Mean coefficients of variation were 12, 5, and 15%, respectively. Statistical analyses were used to conclude that instrumentation error was accountable for only a small portion of the vitamin E variation. Results indicated that a single serum sample assay is an unsatisfactory indicator of vitamin E status in horses. These data have clinical application in the evaluation of horses suspected to be affected with equine degenerative myeloencephalopathy. The large variance of serum total lipids and the lack of correlation of it with serum vitamin E over time preclude the use of vitamin E/serum total lipids ratio in assessing vitamin E status.
Show more [+] Less [-]Serum amyloid A concentrations in cows given endotoxin as an acute-phase stimulant
1989
Boosman, R. | Niewold, T.A. | Mutsaers, C.W.A.A.M. | Gruys, E.
The concentrations of serum amyloid A (SAA) in 4 cows given Escherichia coli endotoxin as an acute-phase stimulant were quantitatively evaluated by use of an indirect micro-ELISA method and compared with other clinical hematologic values. Serum amyloid A concentration changed minimally after intradermal infection of endotoxin. The concentration of SAA was increased 5 hours after IV injection of endotoxin, with maximal concentration after 17 to 20 hours. The increase in SAA concentration coincided with decreasing serum Zn and Fe concentrations; however, Zn and Fe concentrations appeared to be restored when SAA concentration was still maximal. It was concluded that the SAA response of cattle is comparable with that of other species and can be used for monitoring the activity of clinical inflammation and tissue injury.
Show more [+] Less [-]Characterization of the attachment of Treponema hyodysenteriae to Henle intestinal epithelial cells in vitro
1989
Bowden, C.A. | Joens, L.A. | Kelley, L.M.
Properties of the attachment of Treponema hyodysenteriae to Henle intestinal epithelial (HIE 407) cells were examined. The frequency of attachment depended on the motility and viability of the spirochetes. Rabbit hyperimmune and swine convalescent antisera inhibited attachment. Treatment of HIE cells with neuraminidase had no effect on attachment; however, treatment of spirochetes with the enzyme decreased adherence significantly (P = 0.01). Attachment was inhibited by N-acetylneuraminic acid, D-glucuronic acid, and fetuin. Adherence was increased following coincubation with N-acetylglucosamine or yeast mannan. Surface antigens of T hyodysenteriae, isolated by chemical extraction, competitively inhibited adherence. Concentrated T hyodysenteriae culture supernatant fractions inhibited adherence, but concentrated phosphate buffered-saline washings of the spirochete and concentrated uninoculated media did not inhibit adherence. Sialic acid was detected in unwashed T hyodysenteriae and spent culture supernatant fractions in higher concentrations than from washed spirochetes and uninoculated media. It was concluded that the binding adhesins on T hyodysenteriae for cultured HIE cells may contain sialic acid residues.
Show more [+] Less [-]Alterations in bovine neutrophil function during the periparturient period
1989
Kehrli, M.E. Jr | Nonnecke, B.J. | Roth, J.A.
Neutrophils from 8 Holstein heifers were evaluated for function during the periparturient period. Random migration, ingestion of bacteria, superoxide anion production, native (nonenhanced) chemiluminescence, iodination, and antibody-dependent, cell-mediated cytotoxicity by neutrophils were determined. Foremilk samples were evaluated for bacteria. Significant (P less than 0.05) increases in random migration of neutrophils, iodination, and chemiluminescence were evident 2 weeks before parturition and then decreased dramatically by the first week after parturition. These impairments of neutrophil function after parturition may be manifested as a severe cumulative deficit in the native defenses afforded by the neutrophil.
Show more [+] Less [-]Relationship between dietary protein concentration and serum trypsin-like immunoreactivity in dogs
1989
Carro, T. | Williams, D.A.
Serum trypsinogen concentration was studied in 6 adult mixed-breed dogs randomly fed diets containing 6.8, 31.4, or 39.7% protein (dry weight) for 3 weeks each. Blood was collected on days 20, 21, and 22 of each feeding period, and serum trypsinogen concentrations were determined by radioimmunoassay of trypsin-like immunoreactivity (TLI). Mean serum TLI concentrations for each dog fed each diet were compared. A significant (P < 0.05) positive linear relationship (P < 0.02) was determined between serum TLI concentrations and the percentage of dietary protein. Mean serum TLI concentrations for each dog fed all diets ranged from 5.7 to 20.2 microgram/L.
Show more [+] Less [-]Concentration and degree of polymerization of hyaluronate in equine synovial fluid
1989
Saari, H. | Konttinen, Y.T. | Tulamo, R.M. | Antti-Poika, I. | Honkanen, V.
In addition to its well-known physicochemical properties, hyaluronate (HA) has recently been shown to have important biological and pathophysiologic regulatory effects on granulocytes, monocytes, fibroblasts, and endothelial cells, as well as on the healing of wounds and various joint disorders. Many of these effects depend on or are reflected in the concentration and degree of polymerization of HA. Therefore, high-performance liquid chromatography with size-exclusion column was used to characterize the concentration and degree of polymerization of HA in equine synovial fluid (SF). The mean (+/-SD) HA concentration was 0.47 +/- 0.19 mg/ml and there was no difference between control joints and those with positive response to local anesthetic administration (0.61 +/- 0.20 mg/ml vs 0.42 +/-0.17 mg/ml), suggesting that in horses with acute traumatic synovitis causing lameness, HA concentration in SF cannot be used as a marker for the condition. High-performance liquid chromatograms disclosed considerable variation between horses in the degree of polymerization reflected in the peak area to height ratio (mean +/-SD, 3.207 +/- 0.447; range, 2.229 to 3.915), indicating differences in local synthesis, degradation, or mobilization into lymph of SF HA. In addition, the correlation between SF HA concentration and degree of polymerization was 0.760 (P < 0.01; linear regression analysis) suggesting that HA concentration and chain length are independently regulated.
Show more [+] Less [-]Bovine serum and nasal secretion immunoglobulins against Pasteurella haemolytica serotype 1 antigens
1989
Nelson, S.L. | Frank, G.H.
Experimental intranasal inoculation of cattle with Pasteurella haemolytica serotype 1 resulted in a group that shed the bacteria in their nasal secretions (colonized) and a group that did not shed (uncolonized). After inoculation, antibody titers in serum and nasal secretions against the total P haemolytica increased significantly, and the proportion of total antibody against specific P haemolytica antigens changed so that the proportion directed against the 94- and 62-kD antigens increased. Prior to inoculation, the proportion of total antibody in the serum against 94- and 62-kD antigens of P haemolytica was higher in calves that remained uncolonized than in those that became colonized with P haemolytica after exposure. Antibody specificity of serum and nasal secretions differed in the relative amounts directed against each P haemolytica antigen. The specificity against P haemolytica antigens differed between IgG and IgA isotypes of serum and nasal secretions, with IgA being directed against fewer antigens than was IgG.
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