Allan MacKenzie

Evaluation of platelet function in dogs with cardiac disease using the PFA-100 platelet function analyzer

BACKGROUND Cardiac disease has the potential to alter platelet function in dogs. Evaluation of platelet function using the PFA-100 analyzer in dogs of multiple breeds and with a broad range of cardiac conditions would help clarify the effect of cardiac disease on platelets. OBJECTIVES The objective of this study was to assess differences in closure time (CT) in dogs with cardiac disease associated with murmurs, when compared with that of healthy dogs. METHODS Thirty-nine dogs with cardiac murmurs and turbulent blood flow as determined echocardiographically were included in the study. The dogs represented 23 different breeds. Dogs with murmurs were further divided into those with atrioventricular valvular insufficiency (n=23) and subaortic stenosis (n=9). Fifty-eight clinically healthy dogs were used as controls. CTs were determined in duplicate on a PFA-100 analyzer using collagen/ADP cartridges. RESULTS Compared with CTs in the control group (mean+/-SD, 57.6+/-5.9 seconds; median, 56.5 seconds; reference interval, 48.0-77.0 seconds), dogs with valvular insufficiency (mean+/-SD, 81.9+/-26.3 seconds; median, 78.0 seconds; range, 52.5-187 seconds), subaortic stenosis (71.4+/-16.5 seconds; median, 66.0 seconds; range, 51.5-95.0 seconds), and all dogs with murmurs combined (79.6+/-24.1 seconds; median, 74.0 seconds; range, 48.0-187 seconds) had significantly prolonged CTs (P<.01). CONCLUSIONS The PFA-100 analyzer is useful in detecting platelet function defects in dogs with cardiac murmurs, most notably those caused by mitral and/or tricuspid valvular insufficiency or subaortic stenosis. The form of turbulent blood flow does not appear to be an important factor in platelet hypofunction in these forms of cardiac disease.

Measurement of Arginine Kinase Activity in Hemolymph of American Lobsters

Abstract The concentration of arginine kinase (AK) in the hemolymph of lobsters may aid in the identification of muscle diseases in these animals in the same way that creatine kinase activity in blood plasma does in mammalian species. Both manual and automated analysis methods were developed for measuring AK levels in the hemolymph of the American lobster Homarus americanus. A commercial reagent that is used to measure creatine kinase in human blood serum was modified by substituting phospho-L-arginine for creatine phosphate as the substrate to allow measurement of the AK enzyme. Stability of the enzyme in lobster hemolymph plasma was less than 3 h at room temperature (22°C), 6 h when refrigerated (2–5°C), and 24 h when frozen (−20°C). Comparisons were made between the manual and automated protocols using three lobster hemolymph plasma samples. The average recovery in the latter relative to the former was 103%. The automated assay was linear up to 1,940 U/L. Precision studies were conducted using the auto...

Preliminary evaluation of a gel tube agglutination major cross-match method in dogs

BACKGROUND A major cross-match gel tube test is available for use in dogs yet has not been clinically evaluated. OBJECTIVES This study compared cross-match results obtained using the gel tube and the standard tube methods for canine samples. METHODS Study 1 included 107 canine sample donor-recipient pairings cross-match tested with the RapidVet-H method gel tube test and compared results with the standard tube method. Additionally, 120 pairings using pooled sera containing anti-canine erythrocyte antibody at various concentrations were tested with leftover blood from a hospital population to assess sensitivity and specificity of the gel tube method in comparison with the standard method. RESULTS The gel tube method had a good relative specificity of 96.1% in detecting lack of agglutination (compatibility) compared to the standard tube method. Agreement between the 2 methods was moderate. Nine of 107 pairings showed agglutination/incompatibility on either test, too few to allow reliable calculation of relative sensitivity. Fifty percent of the gel tube method results were difficult to interpret due to sample spreading in the reaction and/or negative control tubes. CONCLUSIONS The RapidVet-H method agreed with the standard cross-match method on compatible samples, but detected incompatibility in some sample pairs that were compatible with the standard method. Evaluation using larger numbers of incompatible pairings is needed to assess diagnostic utility. The gel tube method results were difficult to categorize due to sample spreading. Weak agglutination reactions or other factors such as centrifuge model may be responsible.

Comparison of white and red blood cell estimates in urine sediment with hemocytometer and automated counts in dogs and cats

BACKGROUND Therapeutic decisions regarding urinalysis are commonly based on the presence of white and red blood cells. Traditionally, numbers per high-power field are estimated using wet-mount microscopic examination. This technique is not standardized and counts are likely prone to inaccuracy. In addition, differentiation of leukocyte types is not possible. OBJECTIVES The aims of this study were to (1) compare WBC and RBC estimates using wet-mount examination with counts obtained using a hemocytometer, (2) assess if a hematology automated analyzer (Sysmex ST-2000iV/XT) provides reliable WBC and RBC counts in urine comparable to hemocytometer counts, and (3) evaluate air-dried Wright-Giemsa-stained urine drop sediment preparations for the determination of differential leukocyte counts. METHODS WBC and RBC counts were obtained by performing wet-mount estimates, manual hemocytometer counts, and Sysmex automated counts on 219 canine and feline urine samples. Results were correlated using Spearman rank correlation. Air-dried Wright-Giemsa stained sediment drop preparations (n = 215) were examined for differential counts of leukocytes. RESULTS A low but significant association was found between WBC estimates on wet-mount examination and hemocytometer counts (rho = 0.37, P < .01). There was a high and significant association when RBC counts were compared between wet-mount and hemocytometer evaluation (rho = 0.7, P < .01). There was very high and significant interassay correlation between Sysmex data from duplicate samples for what the analyzer classified as WBC (rho = 0.97, P < .01) and RBC (rho = 0.94, P < .01). Low correlations were found between the Sysmex RBC counts and both wet-mount estimates and hemocytometer RBC counts (rho = 0.43, P < .01 and rho = 0.39, P < .01, respectively). Cell preservation in the air-dried sediment preparations was so poor that differential counts could not be performed. CONCLUSION WBC and RBC estimates on wet-mount examination agreed with hemocytometer counts and are therefore considered adequate. The Sysmex ST-2000iV/XT did not provide reliable cell counts under the conditions used.