Comparison of four staining methods for detection of mast cells in equine bronchoalveolar lavage fluid.
Abstract: Mast cells normally are present in equine bronchoalveolar lavage fluid (BALF), but usually represent <2% of all cells in healthy horses. An increased percentage of mast cells has been associated with airway hyperactivity and inflammatory airway diseases, but marked differences are reported between studies in normal and diseased horses. Because an abnormal mast cell count may be of clinical relevance, we compared the ability of a fast Romanowsky method to stain mast cell granules with that of 3 metachromatic stains: automated Romanowsky, May-Grünwald Giemsa, and toluidine blue stains. The BALF cells from 24 horses were studied. A differential cell count was performed blindly on 400 cells. The percentages of mast cells obtained were analyzed by means of repeated-measures analysis of variance and Fischer's PLSD test. The Bland and Altman method was used to assess agreement among stains. The mean percentage of mast cells in BALF was significantly lower with the fast Romanowsky than with the automated Romanowsky, May-Grünwald Giemsa, and toluidine blue stains. With the fast Romanowsky stain, the metachromatic granules of mast cells were not stained, and their identification was based on morphologic criteria. Toluidine blue staining allowed detection of the highest mean percentage of mast cells, but was inadequate for performing a differential cell count on other cell types. In conclusion, fast Romanosky stain may be inadequate for detection of mast cells in equine BALF, whereas automated Romanowsky, May-Grünwald Giemsa, and toluidine blue stains provide metachromatic staining of mast cell granules.
Publication Date: 2006-04-06 PubMed ID: 16594597DOI: 10.1892/0891-6640(2006)20[377:cofsmf]2.0.co;2Google Scholar: Lookup
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- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This study compares four different staining methods for detecting mast cells in horse lung fluid. It finds that a fast Romanowsky method might be less effective for this purpose, while three other metachromatic stains yield better results.
Objectives and Methodology
- The primary aim of the research was to evaluate and compare the effectiveness of four different stains – fast Romanowsky, automated Romanowsky, May-Grünwald Giemsa, and toluidine blue – in detecting mast cells in the bronchoalveolar lavage fluid (BALF) in horses.
- In this experiment, BALF cells from 24 horses were tested using these four staining methods.
- Then, a differential cell count was performed blindly on 400 cells.
- Subsequently, various statistical tests, including repeated-measures analysis of variance and Fischer’s PLSD test, were used to analyze the percentage of mast cells discovered. For assessing the agreement among stains, the Bland and Altman method was employed.
Findings and Interpretation
- Results showed that the mean percentage of mast cells in BALF was significantly lower when stained with the fast Romanowsky method, indicating that it might not be the best choice.
- On the other hand, the other three stains (automated Romanowsky, May-Grünwald Giemsa, and toluidine blue) showed metachromatic staining of mast cell granules, suggesting a better efficiency in mast cell detection.
- The fast Romanowsky stain did not stain the metachromatic granules fo mast cells. Therefore, their identification with this method relied on morphologic criteria alone.
- While toluidine blue staining resulted in the highest mean percentage of mast cells, it was found to be unsatisfactory for conducting a differential cell count on other types of cells.
Conclusion
- To sum up, the research concluded that the fast Romanowsky stain could be inadequate for detecting mast cells in equine BALF.
- In contrast, the other three stains, automated Romanowsky, May-Grünwald Giemsa, and toluidine blue, appear to be more effective by providing metachromatic staining of mast cell granules.
- Future studies might look into enhancing the efficiency of staining methods for not just mast cells but also other types of cells in the BALF.
Cite This Article
APA
Leclere M, Desnoyers M, Beauchamp G, Lavoie JP.
(2006).
Comparison of four staining methods for detection of mast cells in equine bronchoalveolar lavage fluid.
J Vet Intern Med, 20(2), 377-381.
https://doi.org/10.1892/0891-6640(2006)20[377:cofsmf]2.0.co;2 Publication
Researcher Affiliations
- Department of Clinical Sciences, Faculty of Veterinary Medicine, Université de Montréal, Saint-Hyacinthe, Québec, Canada.
MeSH Terms
- Animals
- Bronchoalveolar Lavage Fluid / cytology
- Horses
- Mast Cells / cytology
- Staining and Labeling / methods
- Staining and Labeling / veterinary
Citations
This article has been cited 14 times.- Lakshminarayana S, Sowmya SV, Rao RS. Efficacy of Csaba stain to demonstrate mature and immature mast cells in oral inflammatory lesions: An in-vitro study. J Oral Maxillofac Pathol 2022 Oct-Dec;26(4):483-487.
- 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).
- Grigorev IP, Korzhevskii DE. Modern Imaging Technologies of Mast Cells for Biology and Medicine (Review). Sovrem Tekhnologii Med 2021;13(4):93-107.
- Shawaf T, Almubarak A, Alhumam N, Almathen F, Hussen J. Cytological analysis of tracheal wash and bronchoalveolar lavage fluid in health and respiratory disease in dromedary camels. PeerJ 2021;9:e11723.
- García ER, Barros E, Stransky S, Chávez-Olórtegui C, Freitas MB, Novaes RD, Gonçalves RV. Acidic Phospholipase A2-Peptide Derivative Modulates Oxidative Status and Microstructural Reorganization of Scar Tissue after Cutaneous Injury. Evid Based Complement Alternat Med 2020;2020:8273986.
- Oliveira BM, Pinto A, Correia A, Ferreira PG, Vilanova M, Teixeira L. Characterization of Myeloid Cellular Populations in Mesenteric and Subcutaneous Adipose Tissue of Holstein-Friesian Cows. Sci Rep 2020 Feb 4;10(1):1771.
- Jiang L, Fang P, Septer S, Apte U, Pritchard MT. Inhibition of Mast Cell Degranulation With Cromolyn Sodium Exhibits Organ-Specific Effects in Polycystic Kidney (PCK) Rats. Int J Toxicol 2018 Jul Aug;37(4):308-326.
- Yu Y, Chen Y, Mikael P, Zhang F, Stalcup AM, German R, Gould F, Ohlemacher J, Zhang H, Linhardt RJ. Surprising absence of heparin in the intestinal mucosa of baby pigs. Glycobiology 2017 Jan;27(1):57-63.
- Singh A, Sreedhar G, George J. Incidence of Mast Cells in Gingival and Periapical Inflammation- A Kaleidoscopic Study. J Clin Diagn Res 2016 May;10(5):ZC06-8.
- Houtsma A, Bedenice D, Pusterla N, Pugliese B, Mapes S, Hoffman AM, Paxson J, Rozanski E, Mukherjee J, Wigley M, Mazan MR. Association between inflammatory airway disease of horses and exposure to respiratory viruses: a case control study. Multidiscip Respir Med 2015;10:33.
- Sajadi A, Kafi M, Sharifiyazdi H, Azari M. Rapid leukocyte esterase detection of cytological endometritis in dairy cows. Iran J Vet Res 2025;26(1):78-82.
- Siquier-Dameto G, Iguaran-Pérez A, Gimeno-Beltrán J, Bellia G, Giori AM, Boadas-Vaello P, Verdú E. Subcutaneous Injection and Brush Application of Ovalbumin-Aluminum Salt Solution Induces Dermatitis-like Changes in Mice. J Clin Med 2025 Mar 3;14(5).
- Rasmussen N, Karlsen P, Otten ND, Fjeldborg J, Hansen S. Bilateral bronchoalveolar lavage cytology profiles in a warmblood horse population during a 1-year period. J Vet Intern Med 2024 Jul-Aug;38(4):2391-2398.
- Elieh Ali Komi D, Wöhrl S, Bielory L. Mast Cell Biology at Molecular Level: a Comprehensive Review. Clin Rev Allergy Immunol 2020 Jun;58(3):342-365.
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