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Comparison of methods for estimation of Toxoplasma gondii-specific antibody production in the aqueous humor of cats
1995
Hill, S.L. | Lappin, M.R. | Carman, J.
Intraocular production of Toxoplasma gondii-specific antibody in cats has been estimated by comparing the ratio of T gondii-specific antibody in aqueous humor and serum with the ratio of total immunoglobulins in serum and aqueous humor (Goldmann-Witmer coefficient; aqueous antibody coefficient; C value). It has been proposed that in human beings, comparison of the ratio of T gondii-specific antibody in aqueous humor and serum with the ratio of antibodies against a nonocular pathogen in serum and aqueous humor is more accurate than methods using total immunoglobulin quantification. We developed an ELISA for detection of calicivirus-specific antibodies in the serum and aqueous humor of cats. By evaluating calicivirus-specific antibody concentrations in the aqueous humor of healthy and diseased cats, calicivirus was assessed as a nonintraocular pathogen. The ratio of T gondii-specific antibodies in the aqueous humor and serum and the ratio of calicivirus-specific antibodies in serum and aqueous humor were evaluated as a means of estimating intraocular T gondii-specific antibody production. A field strain of feline calicivirus was isolated, cultured, and purified. A calicivirus-specific IgG ELISA was developed for detection of feline calicivirus-specific IgG in serum and aqueous humor. Calicivirus-specific IgG was measured in the serum and aqueous humor from 3 groups of control cats. Results suggested that calicivirus is a nonintraocular pathogen in cats and that calicivirus IgG detected in aqueous humor is attributable to leakage across a damaged blood-ocular barrier. Intraocular production of T gondii-specific antibodies was estimated, using 2 formulas. The C value was calculated by multiplying the ratio of T gondii-specific IgM or IgG in aqueous humor and serum by the ratio of total immunoglobulins (using the corresponding IgM or IgG class) in serum and aqueous humor. The Ctc value (Toxoplasma-calicivirus Goldmann-Witmer coefficient) was calculated by multiplying the ratio of T gondii-specific IgM or IgG in aqueous humor and serum by the ratio of calicivirus-specific IgG in serum and aqueous humor. Serum and aqueous humor samples were obtained from 41 client-owned cats with uveitis, and T gondii-specific C values and Ctc values were calculated. Toxoplasma gondii-specific IgM or IgG C values of 10 or greater or T gondii-specific IgM or IgG Ctc values of 1 or greater were considered to be suggestive of intraocular T gondii-specific antibody production. Of the 41 cats, 20 (48.7%) had evidence of intraocular production of T gondii-specific antibody on the basis of either an IgM or IgG C value of 10 or greater. A Ctc value could not be calculated in 3 cats because calicivirus-specific IgG was not present in aqueous humor. Of the 38 cats for which Ctc values could be calculated, 25 (65.8%) had evidence of intraocular production of T gondii-specific antibody on the basis of either an IgM or IgG Ctc value of 1 or greater. The C values and Ctc values were in agreement for 75.9% of IgM containing samples and 75% of IgG containing samples. Sensitivity, specificity, predictive value of a positive test result, and predictive value of a negative test result for an IgM or IgG C value, when compared with the corresponding IgM or IgG Ctc value were determined. The results indicate that use of the C value for estimation of intraocular T gondii-specific antibody production will result in 28.6 (IgM) to 50% IgG) false-negative results and 12.5% (IgM and IgG) false-positive results, when compared with the Ctc value.
Show more [+] Less [-]Comparison of a radioimmunoassay (Charm II) test with high-performance liquid chromatography for detection of oxytetracycline residues in milk samples from lactating cattle
1995
Moats, W.A. | Anderson, K.L. | Rushing, J.E. | Wesen, D.P.
A radioimmunoassay test for tetracyclines (Charm II) was compared with high-pressure liquid chromatography (HPLC) for detection of oxytetracycline (OTC) residues in milk samples from individual lactating cows. Oxytetracycline was administered by 1 of 3 routes (IV, IM, or intrauterine) to 21 lactating dairy cows. A total of 292 duplicate milk samples were collected from milkings before and through 156 hours after OTC administration. Concentration of OTC in these samples was determined by use of the Charm II test and an HPLC method with a lower limit of quantitation, approximately 2 ng of OTC/ml. Samples were also classified with respect to presence of OTC residues relative to the FDA safe concentration (less than or equal to 30 ng/ml), using the Charm II (by control point determination) and HPLC methods. There was a significant (P less than or equal to 0.05) difference between test methods in classification of milk samples with respect to presence or absence of OTC at the FDA safe concentration. A total of 48 of the 292 test results (16.4%) did not agree. Using the HPLC test results as the standard with which Charm II test results were compared, 47 false presumptive-violative test results and 1 false presumptive-nonviolative Charm II test result (a sample containing 31 ng of OTC/ml, as evaluated by HPLC) were obtained. The samples with false presumptive-violative Charm II results contained (less than or equal to 30 ng of OTC/ml, as evaluated by HPLC. In some respects, the Charm II test performed appropriately as a screening test to detect OTC residues in milk samples from individual cows. However, the tendency for the test to yield presumptive-violative test results at OTC concentrations lower than the FDA safe concentration (as evaluated by HPLC), suggests that caution should be exercised in using the test as the sole basis on which a decision is made to reject milk. As indicated by the manufacturer, presumptive-violative Charm II test results should be confirmed by additional testing Although not specifically evaluated, the tendency for misclassification of milk samples as presumptive-violative by the Charm II test may or may not occur in commingled milk, compared with milk samples from individual cows.
Show more [+] Less [-]Computed tomographic morphometry of the lumbosacral spine of dogs
1995
Jones, J.C. | Wright, J.C. | Bartels, J.E.
In a 5-year prospective study, computed tomographic (CT) morphometry of the lumbosacral vertebral canal was performed on 42 large-breed dogs (21 controls and 21 dogs with lumbosacral stenosis). Dogs were allotted to 4 groups. Group 1 (n = 13) consisted of cadaver specimens obtained from dogs that died or were euthanatized for reasons unrelated to the spine; group 2 (n = 8) consisted of live dogs with no history of clinical signs related to the spine and with normal neurologic examination findings; group 3 (n = 10) consisted of dogs with surgically confirmed lumbosacral stenosis; and group 4 (n = 11) consisted of dogs with suspected lumbosacral stenosis that were managed conservatively. The CT scans were performed, using 5-mm contiguous slices obtained perpendicular to the vertebral canal, from the midbody of the 5th lumbar vertebra through the caudal endplate of the sacrum (L5-S3). Lumbosacral lordosis was minimized in all dogs by positioning them in dorsal recumbency with the hind limbs flexed. A tuberculin syringe calibration phantom was placed within the scanning field of view, parallel to the axis of the spine. In each dog, 11 CT slice locations within the lumbosacral spine were evaluated. At each slice location, sagittal plane diameter, dorsal plane diameter, and transverse area of the vertebral canal, vertebral body, and calibration phantom were measured, using the CT computer's software programs for distance and area calculation. Window/level settings were constant, and all measurements were made by the same operator (JCJ). Accuracy of calibration phantom CT measurements was 100% for sagittal and dorsal plane diameter and was 85% for transverse area. In control dogs (groups 1 and 2), vertebral canal dimensions were significantly (r greater than or equal to 0.50, P less than or equal to 0.0001) correlated with vertebral body dimensions, but not with dog weight or age. There were no significant differences between group 1 vs group 2, and group 3 vs group 4 for all absolute vertebral canal dimensions and for 5 ratios of vertebral canal to correlated vertebral body dimensions (general linear model for ANOVA). Pooled control dogs (n = 21) and those with lumbosacral stenosis (n = 21) were compared, and significant differences were not identified for absolute canal dimensions. Significant differences between control dogs and those with lumbosacral stenosis were identified in the ratios of vertebral canal transverse area to vertebral body sagittal diameter (P less than or equal to 0.01) and vertebral canal transverse area to vertebral body transverse area (P less than or equal to 0.001). For both these ratios, analysis by slice location identified significant differences (P < 0.05) between pooled groups at the caudal pedicles of L5 and L6. For the ratio of transverse canal area to sagittal vertebral body diameter, differences (P less than or equal to 0.05) also were found at the cranial pedicle of L7. These results indicate that: CT is an accurate method for performing morphometry of the canine lumbosacral spine; vertebral canal dimensions can be corrected for differences in dog size by calculating ratios of vertebral canal to vertebral body dimensions; statistical comparisons, using such corrected vertebral canal dimensions, may reveal differences not evident when absolute vertebral canal dimensions are used; and corrected transverse area of the vertebral canal differs in large-breed dogs with lumbosacral stenosis vs normal controls. Morphometric differences identified at more than 1 vertebral level support a theory that multilevel congenital or developmental stenosis of the lumbosacral vertebral canal may be a predisposing or contributing factor in large-breed dogs with acquired lumbosacral stenosis.
Show more [+] Less [-]Sensitivity and specificity of various serologic tests for detection of Toxoplasma gondii infection in naturally infected sows
1995
Dubey, J.P. | Thulliez, P. | Weigel, R.M. | Andrews, C.D. | Lind, P. | Powell, E.C.
The sensitivity and specificity of various serologic tests for antibodies to Toxoplasma gondii were compared in 1,000 naturally exposed sows, using isolation of viable T gondii as the definitive test. Serum samples obtained from heart blood of 1,000 sows from Iowa were examined for T gondii antibodies by use of the modified agglutination test (MAT), latex agglutination test (LAT), indirect hemagglutination test (IHAT), and ELISA. Toxoplasma gondii was isolated from 170 hearts of 1,000 sows by bioassays in mice and cats. The percentage of samples diagnosed as positive for each of the serologic tests was: MAT = 22.2% (titer >greater than or equal to 1:20), IHAT = 6.4% (titer greater than or equal to 1:64), LAT = 10.4% (titer greater than or equal to 1: 64), and ELISA = 24.1% (OD > 0.360). The sensitivity and specificity of these tests were calculated respectively to be: 82.9 and 90.29% for MAT, 29.4 and 98.3% for IHAT, 45.9 and 96.9% for LAT, and 72.9 and 85.9% for ELISA. The dye test was run at 1:20 dilution on only 893 sera because of bacterial contamination and presence of anticomplement substances. Dye test antibodies were found in 17.8% of the sera, and sensitivity and specificity were 54.4 and 90.8%, respectively. Thus, the MAT had the highest sensitivity among all serologic tests used.
Show more [+] Less [-]Ultrasonography as a method to determine tendon cross-sectional area
1995
Gillis, C. | Sharkey, N. | Stover, S.M. | Pool, R.R. | Meagher, D.M. | Willits, N.
Ultrasonographic cross-sectional area (CSA) measurements of equine superficial digital flexor (SDF) tendon were obtained to determine the feasibility of ultrasonography for CSA measurement of tendon in vivo and in vitro. Ultrasonographic measurements were compared with a more traditional CSA measurement method, ink-blot analysis. In addition, values for ultrasonographic SDF tendon mean echogenicity were obtained in vivo and in vitro. The left forelimb SDF tendons of 23 horses were evaluated ultrasonographically. Cross-sectional images were acquired at 4-cm intervals distal to the base of the accessory carpal bone (DACB) to the level of the proximal sesamoid bones while horses were standing squarely. After euthanasia, the left forelimbs were mounted in a materials testing system (MTS) and loaded under tension to standing load. Ultrasonographic images were again acquired at the same locations. The ultrasonographic images were digitized, and values for ultrasonographic CSA and mean echogenicity were obtained for each level. Immediately after mechanical testing, a 1-cm-thick transverse section of SDF tendon at 12 cm DACB was removed. Three ink blots were prepared from each end of the removed tendon section and digitized. The 6 CSA values were averaged to generate a value for morphologic CSA for each SDF tendon at 12 cm DACB. Standing ultrasonographic tendon CSA at 12 cm DACB was consistently smallest (mean +/- SD CSA = 86 +/- 11 mm2), followed by MTS ultrasonographic CSA (mean, 95 +/- 12 mm2), with ink-blot morphologic CSA being largest (mean, 99 +/- 15 mm2). Comparison of standing and MTS ultrasonographic values at 12 cm DACB revealed a strong positive linear correlation between methods (R2 = 0.74, P = 0.001). Comparison of ink-blot CSA at 12 cm DACB with standing and MTS ultrasonographic CSA revealed strong positive linear correlations (R2 = 0.64, P = 0.001 and R2 = 0.72, P = 0.001, respectively). For ultrasonographic mean echogenicity, standing values insignificantly exceeded MTS values at each level. The authors conclude that ultrasonography is a useful technique for the noninvasive assessment of SDF tendon CSA that can be applied in vivo and in vitro.
Show more [+] Less [-]Comparison of the sensitivity of the caudal fold skin test and a commercial gamma-interferon assay for diagnosis of bovine tuberculosis
1995
Whipple, D.L. | Bolin, C.A. | Davis, A.J. | Jarnagin, J.L. | Johnson, D.C. | Nabors, R.S. | Payeur, J.B. | Saari, D.A. | Wilson, A.J. | Wolf, M.M.
A study to determine and compare the sensitivity of the caudal fold tuberculin test (CFT) and a commercial gamma-interferon (gamma-IFN) assay for diagnosis of bovine tuberculosis was conducted. A dairy herd with approximately a third of the cattle infected with Mycobacterium bovis was chosen for this study. All cattle from this herd were slaughtered, and tissue specimens for bacteriologic culturing and histologic examination were collected. Results of the CFT and gamma-IFN assay were compared with results of bacteriologic culturing and histologic examination to determine test sensitivity. Results were analyzed, using each of the following 4 standards to classify cattle as infected: positive test result by bacteriologic culturing only; histologic examination only; bacteriologic culturing and histologic examination; and bacteriologic culturing or histologic examination. Sensitivity of the CFT ranged from 80.4 to 84.4%, depending on the standard of comparison. Sensitivity of the gamma-IFN assay ranged from 55.4 to 97.1%, depending on the standard of comparison and on the method of interpretation. The CFT was significantly (P < 0.001) more sensitive than the gamma-IFN assay for diagnosis of bovine tuberculosis when the gamma-IFN assay was conducted and interpreted as instructed by the manufacturer. Maximum overall sensitivity was achieved when results of the CFT and gamma-IFN assay were interpreted in parallel.
Show more [+] Less [-]Magnetic resonance arthrography of the scapulohumeral joint in dogs, using gadopentetate dimeglumine
1995
Bree, H van | Ryssen, B. van | Degryse, H. | Ramon, F.
Six scapulohumeral joints (3 normal joints and 3 joints with radiographic evidence of osteochondrosis) underwent conventional magnetic resonance (MR) imaging and MR scapulohumeral arthrography to evaluate delineation of the articular cartilage. The MR arthrography was performed, using 5 ml of 500 micromolar gadopentetate dimeglumine (Gd-DTPA) as a contrast medium. Delineation of normal articular cartilage and cartilage defects was less accurate after intra-articular administration of Gd-DTPA. Therefore, it was concluded that MR arthrography with Gd-DTPA is unrewarding for evaluation of osteochondrosis lesions.
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