Validation of the Sysmex XT‐2000iV hematology system for dogs,cats, and horses. II. Differential leukocyte counts

Background: The Sysmex XT‐2000iV is a laser‐based, flow cytometric hematology system that stains nucleic acids in leukocytes with a fluorescent dye. A 4‐part differential is obtained using side fluorescence light and laser side scatter. Objective: The purpose of this study was to validate the Sysmex XT‐2000iV for determining differential leukocyte counts in blood from ill dogs, cats, and horses. Methods: Blood samples from diseased animals (133 dogs, 65 cats, and 73 horses) were analyzed with the Sysmex XT‐2000iV (Auto‐diff) and the CELL‐DYN 3500. Manual differentials were obtained by counting 100 leukocytes in Wright‐stained blood smears. Results: Leukocyte populations in the Sysmex DIFF scattergram were usually well separated in equine samples, but were not as well separated in canine and feline samples. Correlation among the Sysmex XT‐2000iV, CELL‐DYN 3500, and manual counts was excellent for neutrophil counts (r ≥.97) and good for lymphocyte counts (r ≥.87) for all three species. Systematic differences between the 3 methods were seen for lymphocyte and monocyte counts. The Sysmex reported incomplete differential counts on 18% of feline, 13% of canine, and 3% of equine samples, often when a marked left shift (>10% bands) and/or toxic neutrophils were present. Eosinophils were readily identified in cytograms from all 3 species. Neither the Sysmex nor the CELL‐DYN detected basophils in the 7 dogs and 5 cats with basophilia. Conclusions: The Sysmex XT‐2000iV automated differential leukocyte count performed well with most samples from diseased dogs, cats, and horses. Basophils were not detected. Immature neutrophils or prominent toxic changes often induced errors in samples from cats and dogs.     

Evaluation of piglet blood utilizing the Technicon H*1

The analytic precision of an automated blood analyzer, the Technicon H*1(R), was evaluated utilizing blood samples collected from 20 piglets at 1 and 14 days of age. The effect of storing the blood samples at 4 degrees C for 24 and 48 hours also was determined. Blood samples were analyzed twice on the first day and once on each of the subsequent tow days. Within-sample coefficient of variation was approximately 1% for hemoglobin concentration, erythrocyte count, hematocrit, mean cell volume, erythrocyte distribution width and hemoglobin distribution width (HDW); and approximately 5% for total leukocyte (WBC), neutrophils and lymphocyte counts. Mean HDW and automated differential WBC counts changed during storage to a degree that could be of clinical importance. Manual determination of differential WBC counts were compared with those obtained from the automated analyzer. Results correlated well for neutrophils (r=0.92 in 1-day-old and r=0.93 in 14-day-old piglets, P<0.001) and lymphocytes (r=0.85 in 1-day-old and r=0.93 in 14-day-old piglets, P<0.001). Other WBC values were too low to compare reasonably.     

Feline platelet counting with prostaglandin E1 on the Sysmex XT‐2000iV

Background: The large size of many feline platelets and the high frequency of platelet aggregation often results in falsely low platelet counts in this species. A combination of optical platelet counting to detect even large platelets and the use of prostaglandin E1 (PGE1) to inhibit platelet clumping may increase the accuracy of feline platelet counting. Objective: The objective of this study was to compare platelet counts in feline whole blood samples with and without the addition of PGE1 and using different analytical methods in a clinical setting. Methods: Platelet counts were determined in 10 feline patients in a referral veterinary hospital using 2 sample types (EDTA, EDTA with PGE1) and 2 methods of analysis (optical counting [PLT‐O] and impedance counting [PLT‐I]) on the Sysmex XT 2000 iV analyzer. Results: All PGE1–PLT‐O samples had platelet counts of >200 × 10⁹/L. Mean platelet count using PGE1–PLT‐O (410,256±178 × 10⁹/L) was significantly higher (P<.03) compared with PGE1–PLT‐I (256±113 × 10⁹/L), EDTA–PLT‐O (238±107 × 10⁹/L), and EDTA–PLT‐I (142±84 × 10⁹/L) methods. Depending on the method, platelet counts in 2 to 7 of 10 cats were <200 × 10⁹/L when PGE1‐PLT‐O was not used. A slightly increased platelet count in response to treatment of a feline patient with thrombocytopenia would have been missed without use of PGE1–PLT‐O. Conclusions: Using PLT‐O analysis on EDTA samples containing PGE1 provides higher, and therefore likely more accurate, feline platelet counts in a clinical setting.     

Neutrophilic myeloperoxidase index and mean light absorbance in neonatal septic and nonseptic foals

Background: Two neutrophilic indices reported by the ADVIA 120 Hematology Analyzer, neutrophilic myeloperoxidase index (MPXI), and mean light absorbance (neutrophil X mean [NXM]) have been proposed as indicators of systemic inflammatory disease in horses and of neutrophil activation in coronary ischemic syndromes in people. Objective: The aim of this study was to evaluate NXM and MPXI in healthy, sick nonseptic, and sick septic foals to determine whether conditions likely associated with neutrophil activation result in decreases in these variables. Methods: In this retrospective study, CBC data from 61 neonatal foals presented to the Equine Teaching Hospital of Barcelona were evaluated for correlations between MPXI, NXM, percentage of large unstained cells, neutrophil count, and percentage of band neutrophils. Results obtained in septic (n=32), sick nonseptic (n=22), and healthy foals (n=7) were compared. In addition, results recorded in septic/neutropenic (n=12), septic/non‐neutropenic (n=20), nonseptic/neutropenic (n=8), nonseptic/non‐neutropenic (n=14), and healthy foals (n=7) were also compared. Results: A weak negative correlation was found between MPXI and neutrophil count and between NXM and percentage of band neutrophils. Septic/neutropenic foals had significantly higher MPXI values (median 17.9, minimum–maximum 4.7–42.5) than did septic/non‐neutropenic (1.5, ?24.4 to 22.3), nonseptic/neutropenic (6.6, 0.6–17.9), and nonseptic/non‐neutropenic foals (8.8, ?10.1 to 16.8) but did not differ significantly from controls (12.8, ?8.5 to 20.4). Conclusions: Significant differences in NXM or MPXI were not found when disease groups were compared with controls; however, septic/neutropenic foals had significantly higher median MPXI than other groups of sick foals. Further prospective studies are needed to clarify if this finding is related to decreased neutrophil function or activation in septic/neutropenic foals.     

ASVCP quality assurance guidelines: control of general analytical factors in veterinary laboratories

Owing to lack of governmental regulation of veterinary laboratory performance, veterinarians ideally should demonstrate a commitment to self-monitoring and regulation of laboratory performance from within the profession. In response to member concerns about quality management in veterinary laboratories, the American Society for Veterinary Clinical Pathology (ASVCP) formed a Quality Assurance and Laboratory Standards (QAS) committee in 1996. This committee recently published updated and peer-reviewed Quality Assurance Guidelines on the ASVCP website. The Quality Assurance Guidelines are intended for use by veterinary diagnostic laboratories and veterinary research laboratories that are not covered by the US Food and Drug Administration Good Laboratory Practice standards (Code of Federal Regulations Title 21, Chapter 58). The guidelines have been divided into 3 reports on 1) general analytic factors for veterinary laboratory performance and comparisons, 2) hematology and hemostasis, and 3) clinical chemistry, endocrine assessment, and urinalysis. This report documents recommendations for control of general analytical factors within veterinary clinical laboratories and is based on section 2.1 (Analytical Factors Important In Veterinary Clinical Pathology, General) of the newly revised ASVCP QAS Guidelines. These guidelines are not intended to be all-inclusive; rather, they provide minimum guidelines for quality assurance and quality control for veterinary laboratory testing. It is hoped that these guidelines will provide a basis for laboratories to assess their current practices, determine areas for improvement, and guide continuing professional development and education efforts.     

Hematologic and serum biochemical reference values for the wild Spectacled Caiman, Caiman crocodilus crocodilus, from the Venezuelan plains

Background: Commercial farming of Caiman crocodilus crocodilus has had an impact on the use of this species for meat consumption and the leather industry. Spectacled Caimans comprise part of the South American plains biodiversity. Misinterpretation of laboratory data is a risk owing to the limited hematologic and serum biochemical values available for this species. Objective: The aim of this study was to determine hematologic and serum biochemical values for wild Spectacled Caimans from the Venezuelan plains. Methods: Blood samples were collected form the caudal tail vein of 100 Spectacled Caimans (40 males and 60 females) from the plains located in the State of Apure. Values for RBC count, PCV, hemoglobin concentration, WBC absolute and differential counts, and thrombocyte counts were obtained using manual methods, and RBC indices were calculated. Serum biochemical analysis included measurement of alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, and creatine kinase activities and concentrations of total protein and albumin. Comparisons between sexes were analyzed using the Mann–Whitney test. Results: Reference values for wild Spectacled Caimans were determined. Minor differences in hematologic values, particularly for RBC counts, were found compared with previously published values for this species. Serum biochemical values were similar to those available for other crocodilians. There were no significant differences between males and females. Conclusions: Minor differences between the values obtained for wild Spectacled Caimans and those previously published for this species may be related to differences in methodology and environmental conditions. Availability of hematologic and serum biochemical reference values will be useful for accurate diagnosis and management of disease in this species.