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Veterinary clinical pathology2013; 42(1); 92-98; doi: 10.1111/vcp.12013

Reliability of 400-cell and 5-field leukocyte differential counts for equine bronchoalveolar lavage fluid.

Abstract: Reliable enumeration of mast cells and eosinophils in equine bronchoalveolar lavage (BAL) fluid is important because small increases in the percentages of these cells support the clinical diagnosis of inflammatory airway disease (IAD). Increases in BAL neutrophils also occur with IAD but are not specific due to overlap between IAD and recurrent airway obstruction (RAO). Objective: The objectives of this study were to evaluate the reliability of a standard 400-cell leukocyte differential count and an alternate method evaluating 5 microscopic fields at 500× magnification in equine BAL fluid cytocentrifuged preparations. Methods: BAL samples from 60 horses with and without pulmonary inflammation were evaluated using 400-cell and 5-field leukocyte differential counting methods. Reliability of enumeration of each leukocyte type was assessed by calculating and comparing intraclass correlation coefficients (ICC). Reliability of mast cell enumeration was further evaluated by comparing ICCs of slides with different cell densities. Results: Reliability was higher for all cell types with the 5-field method; however, overall the difference between methods was not statistically significant. Neutrophil reliability was high (ICC > 0.90) with both methods. Adequate reliability (ICC > 0.85) for mast cells was achieved only with the 5-field method on slides with higher cell density. Conclusions: Enumeration of mast cells is unreliable when the standard 400-cell differential counting method is used, whereas the 5-field method on slides with higher cell density reached acceptable reproducibility. Neutrophil percentages were highly reliable with both methods.
© 2013 American Society for Veterinary Clinical Pathology.
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Publication Date: 2013-01-03 PubMed ID: 23289790DOI: 10.1111/vcp.12013Google Scholar: Lookup
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Summary

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The research aimed to evaluate the reliability of different counting methods for assessing the number of mast cells and eosinophils, types of white blood cells, in a fluid sample acquired from horse lungs (bronchoalveolar lavage or BAL). The study compared the reliability of the traditional 400-cell leukocyte differential count method and an alternate five-field method. The findings revealed that the five-field method shows higher reliability, especially for counting mast cells in samples with higher cell density.

Objective and Methods

  • This research was conducted to test the reliability of the 400-cell leukocyte differential counting method and a 5-field method in analyzing equine bronchoalveolar lavage (BAL) fluid preparations. The validity of the count was important as it helped establish the diagnosis of inflammatory airway disease (IAD) in horses. Changes in the number of specific white blood cells, mast cells and eosinophils, provide substantial evidence for diagnosing IAD.
  • The study engaged BAL samples from 60 horses, including those with and without lung inflammation, for the evaluation. The reliability of each leukocyte’s enumeration was determined by calculating intraclass correlation coefficients (ICC).

Results

  • The results revealed that the reliability of counting each cell type was higher with the 5-field method, but this difference was not statistically significant across methods.
  • The percentage of neutrophils was highly reliable, with ICC>0.90, in both the 400-cell and 5-field methods. However, the mast cell enumeration showed an adequate reliability (ICC > 0.85) only in the 5-field method and for slides with higher cell density.

Conclusions

  • The conclusion of this study highlighted that the enumeration of mast cells is unreliable when using the standard 400-cell differential counting method. Instead, the five-field method, especially employed for slides with a higher cell density, provided satisfactory reproducibility. Yet, both methods were excellent for enumerating neutrophils.

Cite This Article

APA
Fernandez NJ, Hecker KG, Gilroy CV, Warren AL, Léguillette R. (2013). Reliability of 400-cell and 5-field leukocyte differential counts for equine bronchoalveolar lavage fluid. Vet Clin Pathol, 42(1), 92-98. https://doi.org/10.1111/vcp.12013

Publication

Veterinary clinical pathology
ISSN: 1939-165X
NlmUniqueID: 9880575
Country: United States
Language: English
Volume: 42
Issue: 1
Pages: 92-98

Researcher Affiliations

Fernandez, Nicole J
  • Department of Veterinary Clinical and Diagnostic Sciences, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB, Canada. fernandn@ucalgary.ca
Hecker, Kent G
    Gilroy, Cornelia V
      Warren, Amy L
        Léguillette, Renaud

          MeSH Terms

          • Animals
          • Bronchoalveolar Lavage Fluid / cytology
          • Eosinophils
          • Female
          • Horse Diseases / diagnosis
          • Horse Diseases / pathology
          • Horses
          • Leukocyte Count / methods
          • Leukocyte Count / veterinary
          • Lung Diseases, Obstructive / diagnosis
          • Lung Diseases, Obstructive / pathology
          • Lung Diseases, Obstructive / veterinary
          • Lymphocytes
          • Macrophages
          • Male
          • Mast Cells

          Citations

          This article has been cited 22 times.
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