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.
Publication Date: 2013-01-03 PubMed ID: 23289790DOI: 10.1111/vcp.12013Google Scholar: Lookup
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- Journal Article
- Bronchoalveolar Lavage
- Cells
- Clinical Pathology
- Clinical Study
- Diagnosis
- Diagnostic Technique
- Disease Diagnosis
- Equine Diseases
- Equine Health
- Inflammation
- Inflammatory Response
- Laboratory Methods
- Leukocytes
- Mast Cells
- Microscopy
- Neutrophils
- Respiratory Disease
- Respiratory Health
- Veterinary Medicine
- Veterinary Research
- White Blood Cells
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
Researcher Affiliations
- Department of Veterinary Clinical and Diagnostic Sciences, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB, Canada. fernandn@ucalgary.ca
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 18 times.- Morini M, Gobbo F, Rinnovati R, Romagnoli N, Peli A, Massarenti C, Spadari A, Pietra M. Bronchoalveolar Lavage Cytology in Severe Equine Asthma: Cytocentrifugated versus Sediment Smear Preparations.. Vet Sci 2023 Aug 16;10(8).
- Bond S, McMullen C, Timsit E, Léguillette R. Topography of the respiratory, oral, and guttural pouch bacterial and fungal microbiotas in horses.. J Vet Intern Med 2023 Jan;37(1):349-360.
- Olave CJ, Ivester KM, Couetil LL, Burgess J, Park JH, Mukhopadhyay A. Effects of low-dust forages on dust exposure, airway cytology, and plasma omega-3 concentrations in Thoroughbred racehorses: A randomized clinical trial.. J Vet Intern Med 2023 Jan;37(1):338-348.
- Basano I, Romolo A, Iamone G, Memoli G, Riccio B, Lavoie JP, Miniscalco B, Bullone M. Giant Multinucleated Cells Are Associated with Mastocytic Inflammatory Signature Equine Asthma.. Animals (Basel) 2022 Apr 20;12(9).
- Slowikowska M, Bajzert J, Miller J, Stefaniak T, Niedzwiedz A. The Dynamics of Circulating Immune Complexes in Horses with Severe Equine Asthma.. Animals (Basel) 2021 Apr 2;11(4).
- Couetil L, Cardwell JM, Leguillette R, Mazan M, Richard E, Bienzle D, Bullone M, Gerber V, Ivester K, Lavoie JP, Martin J, Moran G, Niedźwiedź A, Pusterla N, Swiderski C. Equine Asthma: Current Understanding and Future Directions.. Front Vet Sci 2020;7:450.
- Brankston G, Greer AL, Marshall Q, Lang B, Moore K, Hodgins D, Hennessey JTG, Beeler-Marfisi J. Increased Weekly Mean PM(2.5), and NO(2) Are Associated With Increased Proportions of Lower Airway Granulocytes in Ontario Horses.. Front Vet Sci 2020;7:185.
- Bond SL, Greco-Otto P, MacLeod J, Galezowski A, Bayly W, Léguillette R. Efficacy of dexamethasone, salbutamol, and reduced respirable particulate concentration on aerobic capacity in horses with smoke-induced mild asthma.. J Vet Intern Med 2020 Mar;34(2):979-985.
- Bond SL, Workentine M, Hundt J, Gilkerson JR, Léguillette R. Effects of nebulized dexamethasone on the respiratory microbiota and mycobiota and relative equine herpesvirus-1, 2, 4, 5 in an equine model of asthma.. J Vet Intern Med 2020 Jan;34(1):307-321.
- Bond SL, Hundt J, Léguillette R. Effect of injected dexamethasone on relative cytokine mRNA expression in bronchoalveolar lavage fluid in horses with mild asthma.. BMC Vet Res 2019 Nov 6;15(1):397.
- Davis KU, Sheats MK. Bronchoalveolar Lavage Cytology Characteristics and Seasonal Changes in a Herd of Pastured Teaching Horses.. Front Vet Sci 2019;6:74.
- Varegg MS, Kløverød KM, Austnes MK, Siwinska N, Slowikowska M, Zak A, Niedzwiedz A. The effect of single pretreatment with salbutamol on recovery of bronchoalveolar lavage fluid in horses with suspected or confirmed severe equine asthma.. J Vet Intern Med 2019 Mar;33(2):976-980.
- Rossi H, Virtala AM, Raekallio M, Rahkonen E, Rajamäki MM, Mykkänen A. Comparison of Tracheal Wash and Bronchoalveolar Lavage Cytology in 154 Horses With and Without Respiratory Signs in a Referral Hospital Over 2009-2015.. Front Vet Sci 2018;5:61.
- Léguillette R, Tohver T, Bond SL, Nicol JA, McDonald KJ. Effect of Dexamethasone and Fluticasone on Airway Hyperresponsiveness in Horses With Inflammatory Airway Disease.. J Vet Intern Med 2017 Jul;31(4):1193-1201.
- Niedzwiedz A, Borowicz H, Januszewska L, Markiewicz-Gorka I, Jaworski Z. Serum 8-hydroxy-2-deoxyguanosine as a marker of DNA oxidative damage in horses with recurrent airway obstruction.. Acta Vet Scand 2016 Jun 7;58(1):38.
- Couëtil LL, Cardwell JM, Gerber V, Lavoie JP, Léguillette R, Richard EA. Inflammatory Airway Disease of Horses--Revised Consensus Statement.. J Vet Intern Med 2016 Mar-Apr;30(2):503-15.
- Niedzwiedz A, Jaworski Z. Oxidant-antioxidant status in the blood of horses with symptomatic recurrent airway obstruction (RAO).. J Vet Intern Med 2014 Nov-Dec;28(6):1845-52.
- Niedzwiedz A, Jaworski Z, Tykalowski B, Smialek M. Neutrophil and macrophage apoptosis in bronchoalveolar lavage fluid from healthy horses and horses with recurrent airway obstruction (RAO).. BMC Vet Res 2014 Jan 24;10:29.
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