Reliability of cytological evaluation of mast cells from bronchoalveolar lavage fluid in horses: Intraobserver agreement and mast cell identification

Hansen, Sanni; Fjeldborg, Julie; Hansen, Anders J.; Baptiste, Keith E.

doi:10.1111/eve.13112

Cited by 4 publications

(8 citation statements)

References 23 publications

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“…Increases in the relative percentage of mast cells in BALF may occur more commonly than previously reported . This could be attributable, in part, to laboratory variation in cell staining techniques used or geographical inconsistencies of mastocytic and eosinophilic BALF cytological changes. A small increase in the relative percentage of mast cells within BALF has been associated with AHR in athletic horses .…”

Section: Introductionmentioning

confidence: 98%

The relationship between bronchoalveolar lavage fluid cytology and airway hyper‐reactivity in a population of Australian horses presented for poor performance

et al. 2019

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Objective To investigate the relationship between bronchoalveolar lavage fluid cytology, particularly mast cells, and airway hyper-reactivity in athletic horses presented for poor performance that included a respiratory tract evaluation in two disparate locations in Australia.Design Multi-centre, retrospective and prospective crosssectional studyMethods Eighty four adult horses underwent both pulmonary function testing and histamine bronchoprovocation with a commercial flowmetric plethysmography system. A bronchoalveolar lavage was performed four to twelve hours later. Bronchoalveolar lavage fluid cytology was categorised using two differing classification systems to define mild equine asthma. Statistical analysis was used to assess associations between bronchoalveolar lavage fluid relative inflammatory cell percentages, and airway hyper-reactivity and their associated categorisations.Results Sixty four percent (54/84) of horses displayed airway hyper-reactivity, as defined by PC 35 < 6 mg/ml of histamine. A relative mastocytosis was the most common bronchoalveolar lavage fluid cytological abnormality. Horses with a sole mast cell response of ≥ 5% within their bronchoalveolar lavage fluid displayed airway hyper-reactivity at a lower dose of nebulized histamine than horses with normal bronchoalveolar lavage fluid cytology. Horses with mixed cell responses (relative mast cell percentage > 2% and/or relative neutrophil percentage > 5% and/or eosinophil relative cell percentage ≥ 1%) displayed airway hyperreactivity at a lower dose of nebulized histamine than horses with normal bronchoalveolar lavage fluid cytology. ConclusionIn the Australian context, recently revised increased bronchoalveolar lavage fluid cytology relative cell percentage cut offs appear appropriate for sole mast cell responses. The historical lower cut offs appear to be appropriate for mixed inflammatory cell responses.

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Section: Introductionmentioning

confidence: 98%

The relationship between bronchoalveolar lavage fluid cytology and airway hyper‐reactivity in a population of Australian horses presented for poor performance

et al. 2019

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“…33,34 Only a May-Grünwald-Giemsa staining was included for the BAL cytology, preferably a toluidine blue could have been added for improved mast cell reliability. 4,35 In conclusion, the location of AI in the individual horse varied Compiling to the restrictive reference intervals, one must further have in mind the high risk of a false negative result (4%-25%) when evaluating the BAL sample from only 1 lung. Based on the overall results of this and previous studies, 2,[15][16][17]22 the authors recommend cytological evaluation of both individual lungs for the evaluation of especially mild-moderate asthma.…”

Section: Discussionmentioning

confidence: 99%

“…This theory of the increased amount of dust in the right lung lobe, further conflicts with the findings of a higher BAL mast cell count in the left long lobe compare to the right. The variation in agreement of BAL cytology cell count between studies could also be explained by volume infused for BAL, slide variation, staining method and technique, number of cells counted as well as experience of enumerators 4,5,23 . Most variation will be minimized or even eliminated by applying a standardized BAL cytology protocol, including BAL procedure and subsequently laboratory methods.…”

Section: Discussionmentioning

confidence: 99%

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Bilateral bronchoalveolar lavage cytology profiles in a warmblood horse population during a 1‐year period

Rasmussen,

Karlsen,

Otten

et al. 2024

Veterinary Internal Medicne

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BackgroundBronchoalveolar lavage (BAL) cytology results from 1 lung might not be representative of both lungs.ObjectivesTo determine whether the lung site sampled would influence the horse's BAL cytology profile, and if a pooled BAL sample would be superior with regard to BAL cytology diagnosis in a cohort of healthy and subclinical asthmatic warmblood horses.AnimalsFifty‐nine horses in 2021 and 70 horses in 2022, the follow‐up included 53 of the same in each year.MethodsA cross‐sectional study with follow‐up included BAL cytology samples from individual lungs and from pooled BAL samples. The BAL samples were enumerated and differential cell count were applied to categorize the horses as control or with airway inflammation (AI).ResultsBronchoalveolar lavage mast cell count was higher in left lung compared to right lung (2021; median 1.6 [range, 0.6‐3.3] vs 1.2 [0.7‐1.5] P = .009, 2022; median 3.1 [2.1‐4.2] vs 2.4 [1.7‐3.4], P < .001) and compared to pooled samples (2022; median 2.6 [1.7‐3.7], P < .001). Between year 2021 and 2022, 17 of the horses had changes in BAL cytology from control to AI or vice versa.Conclusions and Clinical ImportancePooled BAL sample was the least reliable for detecting AI, and was not representative of the overall lung condition.

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“…Endoscopic examination including BAL cytology is a valuable tool in the EA diagnostic. 14 The challenges encountered when processing BAL fluid is that it must be performed within a limited time span, using time consuming laboratory techniques 17,[19][20][21] and staining protocols 20,21 in order to ensure adequate quality for microscopic enumeration of cells for differential cell counts. 17 Both time and the temperature of stored BAL samples have been shown to affect cell morphology and increase bacterial growth.…”

Section: Concentration Of Ngal Inmentioning

confidence: 99%

Neutrophil gelatinase‐associated lipocalin as a potential biomarker for equine asthma

Hansen

Otten

Spang-Hanssen

et al. 2023

Equine Veterinary Journal

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BackgroundStudies in people have found neutrophil gelatinase‐associated lipocalin (NGAL) concentrations are increased in asthma and can be used to distinguish between asthma subtypes. NGAL has not yet been investigated in equine asthma (EA).ObjectivesTo investigate the ability of NGAL concentrations in bronchoalveolar lavage (BAL) fluid and serum to distinguish between control horses, horses with mild–moderate EA (MEA) and horses with severe EA (SEA).Study designRetrospective cross‐sectional study.MethodsDetails of endoscopic examination including tracheal mucus score (TMS, scale 0–5) and BAL cytology performed on 227 horses were extracted from records and NGAL concentrations were measured on stored serum and BAL fluid samples. The horses were divided into groups (control group n = 73, MEA n = 98, SEA n = 56) based on clinical signs and BAL cytology results. Differences between groups were evaluated with the Mann–Whitney test and correlation between BAL NGAL, serum NGAL, and BAL cytology were evaluated using Spearman's correlation.ResultsBAL NGAL concentrations were higher in EA than in control horses (median: 25.6 and 13.3 μg/L, respectively, p < 0.001). Concentrations of NGAL in BAL differed between groups, with higher concentrations in MEA than in control horses (median: 18.5 and 13.3 μg/L, respectively, p < 0.001), and higher concentrations in SEA than in MEA horses (median: 54.1 and 18.5 μg/L, respectively, p < 0.001). BAL NGAL concentration differed between horses with TMS ≤2 an >2 (median 15.6 and 21.1 μg/L, respectively, p = 0.004). No differences were found in serum NGAL concentration between any of the groups.Main limitationOnly 66 of the 227 (29%) horses had haematology and serum NGAL measured.ConclusionBAL NGAL concentration differed between control and EA and reflected severity of disease. These results justify further research into the potential of NGAL as a biomarker of EA.

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Reliability of cytological evaluation of mast cells from bronchoalveolar lavage fluid in horses: Intraobserver agreement and mast cell identification

Cited by 4 publications

References 23 publications

The relationship between bronchoalveolar lavage fluid cytology and airway hyper‐reactivity in a population of Australian horses presented for poor performance

The relationship between bronchoalveolar lavage fluid cytology and airway hyper‐reactivity in a population of Australian horses presented for poor performance

Bilateral bronchoalveolar lavage cytology profiles in a warmblood horse population during a 1‐year period

Neutrophil gelatinase‐associated lipocalin as a potential biomarker for equine asthma

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