Objective—To assess and compare the expression of perinuclear antineutrophilic cytoplasmic antibodies (pANCA) in sera obtained from dogs with inflammatory bowel disease (IBD) and dogs with intestinal lymphoma.
Animals—104 dogs with IBD and 23 dogs with intestinal lymphoma.
Procedures—Each ill dog had persistent gastrointestinal signs (> 3 weeks in duration) and absence of response to diet changes or antimicrobial treatments. Gastrointestinal endoscopy was performed in ill dogs to obtain intestinal biopsy specimens for histologic confirmation of IBD or lymphoma. A serum sample was obtained from each ill dog. Neutrophils were isolated from a blood sample from the healthy dog; neutrophil-bearing slides were incubated with serum from each ill dog and examined for expression of pANCA by use of an indirect immunofluorescence technique. Detection of cells that had a perinuclear fluorescence pattern was considered a positive result.
Results—The 2 groups of dogs did not differ with regard to breed and sex but did differ with regard to age. Expression of pANCA was detected in 38 of the 104 (36.5%) dogs with IBD and 4 of the 23 (17.4%) dogs with intestinal lymphoma. Although the frequency of pANCA expression was higher in dogs with IBD, compared with findings in dogs with intestinal lymphoma, the difference was not significant.
Conclusions and Clinical Relevance—Results indicated that circulating pANCA are present in some dogs with IBD or intestinal lymphoma. However, pANCA detection does not seem to be useful for distinguishing dogs with IBD from dogs with intestinal lymphoma.
Objective—To describe the effects of prednisone and acetylsalicylic acid (ASA) on results of thromboelastography in healthy dogs.
Animals—16 male mixed-breed dogs.
Procedures—Dogs were randomly assigned to 3 treatment groups (4 dogs/group) that received prednisone (median dose, 2.07 mg/kg), ASA (median dose, 0.51 mg/kg), or both drugs, PO, every 24 hours from days 0 through 6. Another group received no treatment (control dogs; n = 4). Thromboelastography variables (reaction time, clotting time, α-angle, maximum amplitude [MA], global clot strength, coagulation index, and percentage of clot lysis at 60 minutes [CL60]) were evaluated in blood samples collected (prior to drug administration in treated dogs) on days 0 (baseline), 2, 4, and 6.
Results—Administration of ASA alone did not alter TEG variables. For treatment effect, mean global clot strength was increased in the prednisone and drug combination groups, compared with values for control dogs; MA was also increased in the prednisone and drug combination groups, compared with that of controls. For treatment-by-time effect, median CL60 was increased in the prednisone group on day 6, compared with baseline value in the same dogs and with median CL60 of the control group on day 6. Median CL60 was also increased in the drug combination group on day 6, compared with the baseline value and with that of the control group on day 6.
Conclusions and Clinical Relevance—Prednisone administered at approximately 2 mg/kg/d, PO, for 7 days with or without concurrently administered ASA increased clot strength and decreased clot lysis in healthy dogs.
Objective—To determine the effect of Hct on blood glucose readings of dogs obtained by use of 2 point-of-care (POC) blood glucometers and a laboratory analyzer.
Animals—184 dogs, including 139 Greyhounds.
Procedures—Venous blood samples collected from 184 dogs with a range of Hcts (measured in EDTA-anticoagulated blood) were immediately analyzed with a handheld glucometer specifically developed for veterinary use and a glucometer developed for use in humans. The remainder of each blood sample was placed in fluoride oxalate tubes, and plasma glucose concentration was measured with a laboratory analyzer. Agreement between results for the POC glucometers and laboratory analyzer and effect of Hct on glucometer accuracy was assessed via regression analysis.
Results—Significant differences were detected between results of the glucometers and the reference laboratory analyzer. The Hct affected the correlation between results for the glucometers and the laboratory analyzer. Deviations of the glucometers from the reference interval varied with Hct. The glucometer for veterinary use more closely correlated with the glucose concentration when Hct was within or above its reference interval. The glucometer for use in humans more closely approximated laboratory reference glucose concentrations in anemic dogs.
Conclusions and Clinical Relevance—Hct had a relevant impact on the correlation between whole blood and plasma glucose concentrations in dogs. Significant variations between results obtained with the 2 glucometers could be critical when interpreting blood glucose measurements or selecting a POC glucometer for an intensive care setting and precise glycemic control in critically ill dogs.
Objective—To compare the ease of use and accuracy of 5 feline AB blood-typing methods: card agglutination (CARD), immunochromatographic cartridge (CHROM), gel-based (GEL), and conventional slide (SLIDE) and tube (TUBE) agglutination assays.
Sample Population—490 anticoagulated blood samples from sick and healthy cats submitted to the Transfusion or Clinical Laboratory at the Veterinary Hospital of the University of Pennsylvania.
Procedures—Sample selection was purposely biased toward those from anemic, type B, or type AB cats or those with autoagglutination. All blood samples were tested by use of GEL, SLIDE, and TUBE methods. Fifty-eight samples were also tested by use of CARD and CHROM methods. The presence of alloantibodies in all cats expressing the B antigen as detected by use of any method was also assessed.
Results—Compared with the historical gold-standard TUBE method, good to excellent agreement was achieved with the other typing tests: CARD, 53 of 58 (91% agreement); CHROM, 55 of 58 (95%); GEL, 487 of 490 (99%); and SLIDE, 482 of 487 (99%; 3 samples were excluded because of autoagglutination). Four of the samples with discordant test results originated from cats with FeLV-related anemia.
Conclusions and Clinical Relevance—Current laboratory and in-clinic methods provide simple and accurate typing for the feline AB blood group system with few discrepancies. Retyping after in-clinic typing with the GEL or TUBE laboratory methods is recommended to confirm any type B or AB cats.
Objective—To investigate the influence of diameter of the catheter and blood collection technique on platelet function and variables reflecting secondary hemostasis, physiologic anticoagulation, and fibrinolysis in dogs.
Animals—6 healthy Beagles.
Procedures—Blood samples were collected with 20- and 18-gauge venous catheters immediately after catheters were inserted in a peripheral vein, through a 14-gauge central venous catheter that had been placed via the Seldinger technique in a jugular vein < 30 minutes before sample collection, and through a 13-gauge central venous catheter placed via a catheter-through-the-needle technique < 30 minutes before sample collection (techniques 1 to 4, respectively). Platelet function was assessed in hirudin-anticoagulated whole blood via an impedance-based aggregometer with collagen (0.8, 0.4, 0.2, 0.1, and 0.05 μg/mL) as an inductor. Kaolin-activated thromboelastography variables were determined in citrated whole blood. Prothrombin time, activated partial thromboplastin time, fibrinogen and fibrin D-dimer concentrations, and activity of factor VIII, antithrombin, protein C, and protein S were assessed automatically in citrated plasma.
Results—At 0.05 μg of collagen/mL, the highest median rate of aggregation was observed for collection techniques 2 and 3 with 4.3 (range, 2.5 to 6.5) and 3.7 (range, 2.8 to 8.3) aggregation units/min; however, these values were not significantly different from values for the other collection techniques. Generally, sample collection technique did not have a significant impact on results of coagulation variables investigated.
Conclusions and Clinical Relevance—Various blood collection techniques can be used to obtain samples for coagulation testing.
Objective—To evaluate the influence of treatment with ultralow-dose aspirin (ULDAsp) on platelet aggregation, P-selectin (CD62P) expression, and formation of platelet-leukocyte aggregates in clinically normal dogs.
Animals—18 clinically normal dogs.
Procedures—Studies were conducted before and 24 hours after ULDAsp administration (0.5 mg/kg, PO, q 24 h, for 2 days). Whole blood impedance aggregometry for the assessment of platelet function was performed with sodium citrate–anticoagulated blood and aggregation agonists (ADP at 20, 10, and 5 μmol/L; collagen at 10, 5, and 2 μg/mL). Onset, maximum response, and rate of platelet aggregation were recorded. Flow cytometric assays were configured to detect thrombin-induced CD62P expression and platelet-leukocyte aggregates in EDTA-anticoagulated whole blood. Externalized platelet CD62P and constitutive CD61 (GPIIIa) were labeled with antibodies conjugated to phycoerythrin (PE) and fluorescein isothiocyanate (FITC), respectively. Red blood cell–lysed paraformaldehyde-fixed EDTA-anticoagulated whole blood was dual labeled with CD61-FITC and a panleukocyte antibody (CD18-PE) to characterize platelet-leukocyte aggregates.
Results—ULDAsp significantly delayed platelet aggregation onset with ADP at 20 μmol/L by 54% to 104%, attenuated maximum aggregation with various concentrations of ADP and collagen by ≥ 41%, and slowed aggregation rate with the highest ADP and collagen concentrations by ≥ 39%. Depending on the parameter tested, up to 30% of dogs failed to have an ULDAsp effect. Thrombin stimulation significantly increased CD62P expression in platelets and platelet-leukocyte aggregates, but ULDAsp did not alter basal or thrombin-stimulated CD62P expression.
Conclusions and Clinical Relevance—ULDAsp treatment of clinically normal dogs impaired platelet aggregation in most dogs, but did not influence CD62P platelet membrane expression. (Am J Vet Res 2010;71:1294–1304)
Objective—To use a chromogenic assay to measure tissue factor (TF) activity on the cell surface and in whole cell lysates of feline monocytes in response to treatment with lipopolysaccharide (LPS) and fetal bovine serum (FBS).
Animals—14 healthy cats.
Procedures—Peripheral blood monocytes were isolated via density gradient centrifugation followed by adhesion to plastic. Tissue factor procoagulant activity was measured by use of an assay that detects TF-activated factor X, on the basis of cleavage of a chromogenic TF-activated factor X–dependent substrate. Activity was quantified by comparison with a serially diluted human recombinant TF-activated factor × curve.
Results—The TF procoagulant activity assay was sensitive and specific for TF. Treatment with LPS stimulated TF procoagulant activity on the surface and in whole cell lysates of isolated feline leukocytes. The LPS response in intact cells was dose dependent and cell number dependent and was inhibited by FBS. Monocyte isolation was inefficient, with monocytes comprising a mean of 22% of the isolated cells.
Conclusions and Clinical Relevance—A TF-activated factor X–dependent chromogenic assay that uses human reagents successfully measured surface-expressed and intracellular TF activity of feline monocytes. Treatment with LPS induced TF expression on feline monocytes, but this response was inhibited by FBS. The chromogenic assay was a useful method for measuring TF procoagulant activity in feline cells in vitro and can be used as a research tool to investigate the role of cell-associated TF in thrombotic disorders in cats.
Objective—To compare results reported for blood gas partial pressures, electrolyte concentrations, and Hct in venous blood samples collected from cattle, horses, and sheep and analyzed by use of a portable clinical analyzer (PCA) and reference analyzer (RA).
Animals—Clinically normal animals (24 cattle, 22 horses, and 22 sheep).
Procedures—pH; Pco2; Po2; total carbon dioxide concentration; oxygen saturation; base excess; concentrations of HCO3−, Na+, K+, and ionized calcium; Hct; and hemoglobin concentration were determined with a PCA. Results were compared with those obtained for the same blood sample with an RA. Bias (mean difference) and variability (95% confidence interval) were determined for all data reported. Data were also subjected to analyses by Deming regression and Pearson correlation.
Results—Analysis of Bland-Altman plots revealed good agreement between results obtained with the PCA and those obtained with the RA for pH and total carbon dioxide concentration in cattle, K+ concentration in horses and sheep, and base excess in horses. Except for Na+ concentration and Hct in horses and sheep, correlation was good or excellent for most variables reported.
Conclusions and Clinical Relevance—Data from blood gas and electrolyte analyses obtained by use of the PCA can be used to evaluate the health status of cattle, horses, and sheep. Furthermore, the handheld PCA device may have a great advantage over the RA device as a result of the ability to analyze blood samples on farms that may be located far from urban centers.
Objective—To determine the effect of semen in urine specimens on urine protein concentration measured by means of dipstick analysis.
Sample Population—14 urine samples from 3 adult castrated male dogs and 14 semen samples from 7 adult sexually intact male dogs.
Procedures—Serial dilutions of the whole ejaculate or spermatozoa-free seminal fluid in urine were created, and unaltered and diluted urine samples were analyzed by means of a commercially available dipstick; pH and specific gravity of the samples were also measured. Spermatozoa and WBC counts of the semen samples and protein concentration of the seminal fluid were determined.
Results—Protein concentrations determined by means of dipstick analysis of urine samples to which whole ejaculate (dilutions of 1:1, 1:2, 1:16, 1:64, and 1:256) or seminal fluid (dilutions of 1:1, 1:2, 1:16, and 1:64) had been added were significantly higher than concentrations in unaltered urine samples. All 13 samples to which whole ejaculate was added at a dilution of 1:2 and 10 of 12 samples to which seminal fluid was added at a dilution of 1:2 were positive for blood on dipstick analysis. There was no significant linear correlation between spermatozoa or WBC count of the semen sample and protein concentration of the spermatozoa-free seminal fluid.
Conclusions and Clinical Relevance—Results suggested that regardless of whether spermatozoa were present, semen contamination could result in false-positive results for protein and blood during dipstick analysis of urine samples from sexually intact male dogs.
Objective—To evaluate agreement between 2 portable triglyceride meters and a veterinary laboratory for measurement of blood triglyceride concentrations in dogs and evaluate effects of Hct and blood volume analyzed.
Sample Population—97 blood samples collected from 60 dogs.
Procedures—Triglyceride concentrations were measured in blood by use of 2 meters and compared with serum triglyceride concentrations determined by a veterinary laboratory. Within- and between-day precision, accuracy, and effects of blood volume and Hct were analyzed.
Results—Accuracy of both meters varied with triglyceride concentration, although both accurately delineated dogs with triglyceride concentrations < 180 mg/dL versus ≥ 180 mg/dL. One meter had results with excellent overall correlation with results of the standard laboratory method, with a concordance correlation coefficient of 0.94 and mean difference of 20.3 mg/dL. The other meter had a good overall concordance correlation coefficient of 0.86 with a higher absolute mean difference of −27.7 mg/dL. Results were only affected by blood volume; triglyceride concentrations determined via both meters were significantly lower when 7 μL of EDTA-anticoagulated blood was used, compared with larger volumes.
Conclusions and Clinical Relevance—1 meter had greater accuracy in the range of 140 to 400 mg/dL and was therefore well suited to detect hypertriglyceridemia. The other meter was accurate with triglyceride values < 140 mg/dL and yielded results similar to those of the veterinary laboratory in the range of 140 to 400 mg/dL, therefore being suitable for determination of triglyceride concentrations in nonfed dogs and dogs with mildly high concentrations.