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.     

Hematologic values in calves during the first 6 months of life

BACKGROUND: Age-related changes in hematologic values are known to occur in many species. Few published studies include repeated measurements of hematologic parameters in calves during the first months of life. OBJECTIVE: The aim of the present study was to monitor hematologic values by sequential measurements from birth to 6 months of age in 15 healthy calves of the Norwegian Red breed, and compare the results to reference intervals for adult, lactating dairy cows. METHODS: Fifteen clinically healthy calves were sampled every week during the first 5 weeks of life and every month thereafter until 6 months of age. Hematologic values were measured using the ADVIA 120 hematology system. Reference intervals were determined for 75 healthy adult cows of the same breed. RESULTS: Compared with adult reference intervals, the MCV was lower and the RBC count was higher in calves throughout the investigation period. Hemoglobin concentration stayed largely within the adult reference interval. Mean MCHC was lower than adult values for 5 weeks, then increased and reached adult values by weeks 10-12. The mean lymphocyte count for calves reached adult reference values at weeks 6-8, and the mean monocyte count increased steadily until weeks 14-16. For most leukocytes, interindividual variation was larger during the first 5-8 weeks of life. The mean platelet count for calves was higher than the adult reference interval until weeks 19-21 of age. CONCLUSIONS: Age-specific reference intervals for calves from birth to 6 month of age are needed for RBC count, MCV, MCHC, red cell distribution width, and platelet and lymphocyte counts.     

Artifactual changes in canine blood following storage, detected using the ADVIA 120 hematology analyzer

BACKGROUND: Artifactual changes in blood may occur as a consequence of delayed analysis and may complicate interpretation of CBC data. OBJECTIVE: The aim of this study was to characterize artifactual changes in canine blood, due to storage, using the ADVIA 120 hematology analyzer. METHODS: Blood samples were collected into EDTA from 5 clinically healthy dogs. Within 1 hour after blood sample collection and at 12, 24, 36 and 48 hours after storage of the samples at either 4 degrees C or room temperature (approximately 24 degrees C), a CBC was done using the ADVIA 120 and multispecies software. A linear mixed model was used to statistically evaluate significant differences in values over time, compared with initial values. RESULTS: The HCT and MCV were increased significantly after 12 hours of collection at both 4 degrees C and 24 degrees C, and continued to increase through 48 hours. The MCHC initially decreased significantly at 12-24 hours and then continued to decrease through 48 hours at both temperatures. Changes in HCT, MCV, and MCHC were greater at 24 degrees C than at 4 degrees C at all time points. A significant increase in MPV and a decrease in mean platelet component concentration were observed at all time points at 24 degrees C. Samples stored at 24 degrees C for 48 hours had significantly higher percentages of normocytic-hypochromic RBCs, and macrocytic-normochromic RBCs, and lower platelet and total WBC counts. CONCLUSIONS: Delayed analysis of canine blood samples produces artifactual changes in CBC results, mainly in RBC morphology and platelet parameters, that are readily detected using the ADVIA 120. Refrigeration of specimens, even after 24 hours of storage at room temperature, is recommended to improve the accuracy of CBC results for canine blood samples.     

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.     

Reticulocyte indices in a canine model of nutritional iron deficiency

BACKGROUND: Reticulocyte indices, especially reticulocyte hemoglobin content (CH retic), have shown promise as markers of iron deficiency (ID), but there have been no prospective investigations of reticulocyte indices in experimental models of ID. OBJECTIVES: The objective of this study was to compare reticulocyte indices with conventional hematologic and biochemical indices as markers of ID in dogs. METHODS: Iron deficiency was induced in 7 dogs by feeding an iron-deficient diet, and corrected by restoring dietary iron and by giving iron parenterally. Blood samples were collected at weekly intervals. Results of hematologic and biochemical tests were compared using t-tests and receiver operator characteristic (ROC) curve analysis. RESULTS: Comparing mean values on days 0 and 35, by which time hemoglobin concentration decreased to 90% of baseline in all dogs, % Macro retic, % Hypo retic, % Low CH retic, and % High CH retic differed by greater than 3-fold, whereas no conventional hematologic or biochemical indices differed by as much as 2-fold. Comparing conventional hematologic and reticulocyte indices by ROC curve analysis using 4 different biochemical diagnostic criteria of ID, CH retic, % Hypo retic, % Low CH retic, and % High CH retic had higher areas under the curve (AUC) than either MCV conv or MCHC conv according to all 4 criteria, and % Macro retic and MCV retic had higher AUC values according to 3 of 4 criteria (differences were not always statistically significant). CONCLUSIONS: Results of this study support the value of reticulocyte indices in the diagnosis and monitoring of ID in dogs. Species similarities in the pathophysiology and hematologic manifestations of ID suggest these findings also may be relevant to ID in people.