Comparison of methods for determining platelet numbers and volume in Cavalier King Charles spaniels

OBJECTIVE: The purpose of this study was to compare platelet concentration in cavalier King Charles spaniels (CKCS) measured by different methods commonly used in veterinary hospitals and commercial laboratories. METHODS: Blood samples obtained from 41 CKCS [corrected] were analysed by impedance cell counter, laser cell counter and microscopic estimation. Quantitative buffy coat analysis was performed only on 17 samples, selected from CKCS [corrected] that had low platelet counts detected by cell counters. Platelet counts, platelet estimations and platelet parameters using these different methods were compared. RESULTS: The median platelet number was lower when estimated using impedance cell counter (1363x10(9)/I) with respect to laser cell counter (1723x10(9)/I), microscopic estimation (238x10(9)/I) [corrected] or quantitative buffy coat analyser (292x10(9)/I) [corrected] (P<0.01). Although impedance cell counter, laser cell counter and microscopic estimation were positively correlated, there was no acceptable agreement among methods. CKCS [corrected] with macrothrombocytes in blood smears had significantly lower counts on impedance cell counter, laser cell counter and microscopic estimation. The percentages of CKCS [corrected] with platelet count < 100x10(9)/I [corrected] were 34.1 per cent (impedance cell counter), 26.8 per cent (laser cell counter), 22.0 per cent (microscopic estimation) (not statistically different) and 5.8 per cent (quantitative buffy coat analyser) (P<0.05). CLINICAL SIGNIFICANCE: CKCS [corrected] with macrothrombocytosis have low platelet counts on impedance cell counters, laser cell counters and microscopic estimation. CKCS [corrected] with low platelet counts may have a normal platelet crit detected by a quantitative buffy coat analyser and thus a normal circulating platelet mass.     

Comparison between manual and automated total nucleated cell counts using the ADVIA 120 for pleural and peritoneal fluid samples from dogs,cats, horses,and alpacas

Background: Analysis of body fluids includes an estimate of total nucleated cell count (TNCC). Automated methods may enhance the accuracy and timeliness of TNCC results. Objective: The purpose of this report was to assess the ability of the ADVIA 120 hematology analyzer to accurately count nucleated cells in pleural and peritoneal fluids from animals, compared with manual counts. Methods: Pleural and peritoneal fluids submitted in EDTA tubes to our laboratory over a 17‐month period were used in the study. TNCC/μL was determined by a manual method, using a hemocytometer, and by an automated method, using the ADVIA 120. Correlation of results was determined by Passing‐Bablok regression, Bland–Altman plots, and Pearson correlation analysis. Results: Samples from dogs (n=36), cats (n=36), horses (n=59), and alpacas (n=11) were analyzed. High correlation in TNCC between methods was found for peritoneal fluid (n=93, r=.959), pleural fluid (n=49, r=.966), and all fluids combined (n=142, r=.960) (P<.001). Variation between methods was greater in samples with TNCCs<1000/μL (r=.62, P<.001). The ADVIA systematically overestimated the number of cells in all fluid samples by 95 cells/μL (confidence interval=19.2–190.5/μL). Conclusion: The ADVIA 120 reliably determines TNCC in pleural and peritoneal effusions and can be recommended for routine veterinary laboratory analysis.     

Evaluation of flow cytometric and automated methods for detection of activated platelets in dogs with inflammatory disease

OBJECTIVE: To evaluate platelet surface-associated P-selectin, mean platelet component concentration (MPC), mean platelet component distribution width (MPCDW), mean platelet volume (MPV), and platelet distribution width (PDW) for detection of activated platelets in dogs with septic and nonseptic inflammatory disease. ANIMALS: 20 healthy dogs and 20 dogs with septic and nonseptic inflammatory disease. Procedures-Platelet surface-associated P-selectin (expressed as the median fluorescence intensity [MFI] of the platelet population), MPC, MPCDW, MPV, and PDW were determined in 20 healthy adult dogs, and reference ranges were calculated. These parameters were also determined in 11 dogs with nonseptic and 9 dogs with septic inflammatory disease and evaluated to determine which parameters were useful for detection of activated platelets. Results-12 dogs with inflammatory disease had P-selectin greater than the upper limit of the reference range, whereas 16 dogs with inflammatory disease had MPC lower than the lower limit of the reference range. All dogs in which P-selectin was greater than the upper limit of the reference range had MPC lower than the lower limit of the reference range. The correlation coefficient for P-selectin and MPC was 0.62. Differences in the MPCDW, MPV, and PDW in most dogs with inflammatory disease (compared with healthy dogs) were found; however, the correlation coefficients for P-selectin and MPCDW, MPV, and PDW were low. CONCLUSIONS AND CLINICAL RELEVANCE: Platelet surface-associated P-selectin and MPC appeared to be useful to detect activated platelets in most dogs with septic and nonseptic inflammatory disease.     

Comparison of two automated spectrophotometric methods for ceruloplasmin measurement in pigs

Ceruloplasmin (Cp) determination could provide an objective measure of the health status of an animal and could be used as marker of animal health and welfare (Skinner, 2001) (Skinner, J. G., 2001. Special report. International standardization of acute phase proteins. Veterinary Clinical Pathology 30, 2-7.) but only manual methods have been reported to determine Cp concentrations in pigs (McCosker, 1961; Toussaint et al., 1995; Eckersall et al., 1996) (McCosker, P. J., 1961. Paraphenylenediamine oxidase activity and copper-levels in mammalian plasmas. Nature. 190, 887-889; Toussaint, M. J. M., Van Ederen, A. M., Gruys, E., 1995. Implication of clinical pathology in assessment of animal health and in animal production and meat inspection. Comparative Haematology International 5, 149-157; Eckersall, P. D., Saini, P. K., McComb, C., 1996. The acute phase response of acid soluble glycoprotein, alpha(1)-acid glycoprotein, ceruloplasmin, haptoglobin and C-reactive protein, in the pig. Veterinary Immunology and Immunopathology 51, 377-385). In the present study two automated methods based on the use of two different substrates for the determination of serum ceruloplasmin in pigs were developed, evaluated and compared. Both methods showed a good precision and detection limits, with no signs of inaccuracy and could be applied to biochemical autoanalyzers usually found in clinical laboratories using only minimal amounts of serum. Additionally the behaviour of Cp in experimental and clinical situations was studied showing an increase of around two fold after turpentine administration and significantly higher values in cases of pigs with inflammatory conditions when compared with healthy pigs.     

Prevalence of low automated platelet counts in cats: comparison with prevalence of thrombocytopenia based on blood smear estimation

Abstract: True thrombocytopenia is uncommon in cats; however, low platelet counts frequently are found using automated cell counters. Although this discrepancy is a well known problem, the prevalence of low automated platelet counts in feline blood samples has not been documented. We retrospectively compared the prevalence of low automated platelet counts with low blood smear-estimated platelet counts in feline blood samples. Results of blood sample analysis from 359 cats during a 1-year period at the University of Glasgow Veterinary Haematology Laboratory were examined. Smear estimates of platelet number were done in those cases in which records did not indicate adequate platelet numbers. Platelet counts obtained with an impedance counter (Minos Vet, Abx Hematologie) were <200×10⁹ cells/L in 256 samples (71%) and <50×10⁹ cells/L in 43 samples (12%). However, based on estimation of platelet numbers from blood smears, only 11 samples (3.1%) had platelet counts of <200×10⁹ cells/L and 9 samples (2.5%) had counts of <50×10⁹ cells/L. Four cats with thrombocytopenia estimated by blood smear evaluation had clinical signs of a bleeding disorder. Disorders associated with thrombocytopenia included neoplasia, cytotoxic chemotherapy, and infectious diseases. There was no evidence that delay due to mailing of samples was associated with lower automated platelet counts than would have been obtained on the day of sampling. The high prevalence of apparent thrombocytopenia in automated platelet counts was attributed to a combination of platelet aggregation and the impedance method of cell differentiation by size. Vigilance and careful examination of blood smears is required to identify the few cats with true thrombocytopenia.     

Comparison of various methods for estimating body fat in dogs

Obesity is considered one of the most common forms of malnutrition occurring in dogs. Laboratory methods of evaluation of body composition in live dogs have included dual-energy X-ray absorptiometry (DEXA) and deuterium oxide (D(2)O) dilution. Clinical methods of evaluation include assigning a body condition score (BCS) based on visual observation, palpation, and morphometric measurements. This study used these four methods to evaluate 23 healthy, adult, client-owned dogs. Good correlation (coefficient of determination [r(2)]=0.78) was found between measurements of percent body fat (%BF) determined by the D(2)O dilution method and the DEXA scan. Percent body fat can also be estimated using BCS (r(2)=0.92 comparison with DEXA) or by using morphometric measurements with simple calculations (r(2)=0.92 comparison with DEXA).     

Comparison of three radiographic methods for diagnosis of hip dysplasia in eight-month-old dogs.

OBJECTIVE: To compare the accuracy of the extended-hip radiographic (EHR) score, the distraction index (DI), and the dorsolateral subluxation (DLS) score for identifying hip dysplasia in dogs at 8 months of age. DESIGN: Cohort study ANIMALS: 129 Labrador Retrievers, Greyhounds, and Labrador Retriever-Greyhound crossbreds. PROCEDURE: Radiography was performed when dogs were 8 months of age. Dogs were euthanatized at 8 to 36 months of age; hip dysplasia was diagnosed at the time of necropsy on the basis of results of a gross examination of the articular cartilage of the hip joints for signs of osteoarthritis. RESULTS: The EHR score, DI, and DLS score at 8 months of age were all significantly correlated with degree of cartilage degeneration at necropsy. Sensitivity and specificity of using EHR score at 8 months of age to diagnose hip dysplasia (scores > 3 were considered abnormal) were 38 and 96%, respectively; sensitivity and specificity of using DI (values > 0.7 were considered abnormal) were 50 and 89%; and sensitivity and specificity of using DLS score (scores < 55% were considered abnormal) were 83 and 84%. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that specificities of the 3 methods for diagnosing hip dysplasia in dogs at 8 months of age were similar. However, the DLS score had higher sensitivity, indicating that there were fewer false-negative results.     

Clinical application of reticulocyte counts in dogs and cats.

Reticulocytes are anucleate immature red blood cells that contain a network of RNA, organelles, and mitochondria, which stain with supravital dyes. Both aggregate and punctate reticulocytes are present in domestic cats, and aggregate reticulocytes are used to assess the degree of regeneration in anemic dogs and cats. Multiple factors influence the degree of regenerative response to anemia. These factors include time of reticulocyte measurement, concurrent diseases, species, and ongoing therapy. Although many automated systems for reticulocyte enumeration exist, manual counts remain the gold standard in veterinary medicine.     

Triggers for prophylactic use of platelet transfusions and optimal platelet dosing in thrombocytopenic dogs and cats.

Prophylactic platelet transfusions are frequently given to human patients with hypoproliferative thrombocytopenia. For several decades, the most common transfusion trigger was 20,000/microL, but the trend is now to use 10,000/microL in the absence of other risk factors for bleeding. This trigger seems to reduce the number of transfusions without increasing the risk of severe bleeding. Most studies involved in establishing platelet transfusion policies have involved patients with acute leukemia, with fewer studies involving patients undergoing hematopoietic stem cell transplantation or aggressive chemotherapy for other cancers and patients with aplastic anemia. In the presence of other risk factors for spontaneous bleeding, 20,000/microL is still considered an appropriate trigger. The trigger for prophylactic transfusion before surgery has not undergone the same recent scrutiny as has the trigger for spontaneous bleeding. The recommendation remains to raise the platelet count to 50,000 to 100,000/microL if possible, although it is recognized that surgery and other invasive procedures have been performed at lower platelet counts without major bleeding. Prophylactic transfusion is not used in disorders of platelet consumption and destruction to prevent spontaneous bleeding but is used before surgery. Because of the comparative lack of experience with platelet transfusion in veterinary medicine, it is difficult to make generalizations for dogs and cats. Using the guidelines established for therapeutic and prophylactic transfusion of human patients is a reasonable starting point, however. A therapeutic transfusion policy is suggested in the veterinary setting provided that the patient can be closely observed for critical bleeding and a prompt transfusion can be given. This policy should ultimately reduce the overall number of platelet transfusions given to hospital patients. If an animal cannot be closely observed or the ability to transfuse on demand is limited, prophylactic transfusion is recommended. The triggers for initiating a platelet transfusion in dogs are extrapolated from human data; these values are lower by 50% for cats. Because of the imprecision of platelet counting at low values, platelet counts must always be interpreted in conjunction with clinical signs of hemorrhage. If platelet-rich plasma or platelet concentrate is available, a dose of 1 platelet unit per 10 kg is recommended, although resources may dictate a smaller dose. This will raise the recipient platelet count by a maximum of about 40,000/microL. Assuming a trigger of 10,000/microL, a transfusion will probably be required approximately every 3 days. It must be remembered that the frequency of platelet transfusions may be greater in the presence of factors accelerating platelet loss or destruction. If fresh whole blood is used, a rule of thumb is to transfuse 10 mL/kg, which will raise the recipient platelet count by a maximum of approximately 10,000/microL. Daily transfusions or transfusions every other day will probably be required.     

Comparison of some serological methods and coproscopic examinations for diagnosis of Giardia spp. invasion in dogs

Giardiasis was detected in 53.5% of dogs examined by FASTest Giardia Strip for use in dogs. Using the ProSpecT Giardia EZ Microplate Assay 52.2% of these results was confirmed. Cysts of Giardia spp. were found only in 6.5% of samples of feces examined by flotation or decantation techniques. The examinations confirmed problems with coproscopic diagnosis of giardiasis in dogs. They confirmed the greater usefulness of FASTest Giardia Strip for immunodiagnostic of giardiasis in carnivores.