Topic:Bronchoalveolar Lavage
Bronchoalveolar lavage (BAL) is a diagnostic procedure used to collect samples from the lower respiratory tract of horses. The technique involves the introduction of a sterile fluid into the bronchi and alveoli, which is then retrieved for analysis. BAL is utilized to assess the cellular and microbiological composition of the lower airways, aiding in the diagnosis of respiratory conditions such as inflammatory airway disease, recurrent airway obstruction, and infections. The procedure provides valuable insights into the type and extent of inflammation or infection present in the respiratory system. This page compiles peer-reviewed research studies and scholarly articles that explore the methodology, applications, and clinical relevance of bronchoalveolar lavage in equine respiratory health.
Effects of low-dust forages on dust exposure, airway cytology, and plasma omega-3 concentrations in Thoroughbred racehorses: A randomized clinical trial. Racehorses commonly develop evidence of mild asthma in response to dust exposure. Diets deficient in omega-3 polyunsaturated fatty acids (Ω-3) might exacerbate this response. Objective: To compare dust exposure, bronchoalveolar lavage fluid (BALF) cytology, and plasma Ω-3 and specialized pro-resolving mediators (SPM) concentrations amongst racehorses fed dry hay, steamed hay, and haylage. Methods: Forty-three Thoroughbred racehorses. Methods: Prospective clinical trial. Horses were randomly assigned to be fed dry hay, steamed hay, or haylage for 6 weeks. Measures of exposure to dust in the...
Free Swimming and Exercise-Induced Pulmonary Hemorrhage in Endurance Horses: A Preliminary Study. Swimming is used for rehabilitation of musculoskeletal injuries and for conditioning to improve equine fitness. However, there are anecdotal reports that suggest that tethered swimming can induce epistaxis, likely secondary to exercise-induced pulmonary hemorrhage (EIPH). The objective of this observational, prospective study was to investigate if EIPH occurs during intensive free-swimming training sessions (5 × 70m) using 15 endurance horses. On tracheo-bronchoscopic evaluations following swimming, low grade mucus scores were observed, but no tracheal blood was observed. Bronchoalveolar la...
Impact of Lower Airway Inflammation on Fitness Parameters in Standardbred Racehorses. Mild-moderate equine asthma (MEA) is a common respiratory disorder among racehorses, characterized by lower airway inflammation. Although it is generally agreed that MEA causes poor performance, contrasting results have been reported about the effects of tracheal mucus and the leukocyte populations of the bronchoalveolar lavage (BAL) on performance. This study aims to investigate the associations between airway inflammation and fitness parameters measured during an incremental treadmill test on Standardbred racehorses. For this purpose, the clinical records of 116 Standardbreds subjected to a ...
Molecular detection of equid herpesvirus in bronchoalveolar lavage fluid from asymptomatic horses in Southern Brazil. Molecular approaches to diagnose respiratory viruses have provided an opportunity for early and subclinical pathogen detection, particularly in samples from the upper respiratory tract. This study aimed to investigate the presence of herpesviruses, particularly equid herpesvirus (EHV)-2 and EHV-5, in samples from the lower respiratory tract of healthy racehorses from Southern Brazil. Unassigned: Samples from the lower respiratory tract (i.e., bronchoalveolar lavage fluid [BALF]) were assessed by video endoscopy, cytological evaluation of BALF, and tracheal aspirates (TA), along with quantitati...
Cytologic scoring of equine exercise-induced pulmonary hemorrhage: Performance of human experts and a deep learning-based algorithm. Exercise-induced pulmonary hemorrhage (EIPH) is a relevant respiratory disease in sport horses, which can be diagnosed by examination of bronchoalveolar lavage fluid (BALF) cells using the total hemosiderin score (THS). The aim of this study was to evaluate the diagnostic accuracy and reproducibility of annotators and to validate a deep learning-based algorithm for the THS. Digitized cytological specimens stained for iron were prepared from 52 equine BALF samples. Ten annotators produced a THS for each slide according to published methods. The reference methods for comparing annotator's and al...
Quantitative Transcriptome Analysis of Purified Equine Mast Cells Identifies a Dominant Mucosal Mast Cell Population with Possible Inflammatory Functions in Airways of Asthmatic Horses. Asthma is a chronic inflammatory airway disease and a serious health problem in horses as well as in humans. In humans and mice, mast cells (MCs) are known to be directly involved in asthma pathology and subtypes of MCs accumulate in different lung and airway compartments. The role and phenotype of MCs in equine asthma has not been well documented, although an accumulation of MCs in bronchoalveolar lavage fluid (BALF) is frequently seen. To characterize the phenotype of airway MCs in equine asthma we here developed a protocol, based on MACS Tyto sorting, resulting in the isolation of 92.9% pur...
Nebulized glycosylated caffeic acid phenylether ester attenuation of environmental particulate-induced airway inflammation in horses. The objective of this study was to determine the extent that nebulized glycosylated caffeic acid phenylether ester-4-O-alpha-D-glucopyranoside (G-CAPE) attenuates particulate-induced airway inflammation in healthy horses. Our hypothesis was that nebulization with G-CAPE would result in improved respiratory scores, higher arterial oxygen partial pressure, and less inflammatory airway infiltrates in horses with induced airway inflammation, compared with untreated controls. Five healthy adult horses were housed inside a climate controlled, closed barn on straw bedding and fed moldy grass hay for...
Long-Read Transcriptome of Equine Bronchoalveolar Cells. We used Pacific Biosciences long-read isoform sequencing to generate full-length transcript sequences in equine bronchoalveolar lavage fluid (BALF) cells. Our dataset consisted of 313,563 HiFi reads comprising 805 Mb of polished sequence information. The resulting equine BALF transcriptome consisted of 14,234 full-length transcript isoforms originating from 7017 unique genes. These genes consisted of 6880 previously annotated genes and 137 novel genes. We identified 3428 novel transcripts in addition to 10,806 previously known transcripts. These included transcripts absent from existing genome...
Effect of the Administration of a Nutraceutical Supplement in Racehorses with Lower Airway Inflammation. Mild−moderate equine asthma (MEA) is a chronic inflammatory disorder of the lower airways of the horse, characterized by tracheal mucus accumulation, cough and poor performance. The therapeutic approach is based on pharmacological treatment and environmental management. Moreover, the efficacy of the administration of antioxidant molecules has been reported. The aim of the present study was to evaluate the effect of the administration of a commercial nutraceutical supplement, composed of several herbal extracts, on lower airway inflammation in racehorses. Twelve Thoroughbreds affected by MEA ...
Diagnostic Value of Tracheal Wash Cytology for Monitoring Exercise-Induced Pulmonary Hemorrhage in Thoroughbred Racehorses. Lower airway cytology has been proposed as a complementary diagnostic method to confirm the presence and quantifying the severity of exercise-induced pulmonary hemorrhage (EIPH). Bronchoalveolar lavage is usually preferred over tracheal wash (TW), yet the need for sedation imposes as a limitation for active sport horses in addition to be a highly invasive technique. This study aims to evaluate the diagnostic value of TW with Total Hemosiderin Score (THS) for monitoring EIPH in active Thoroughbred racehorses. A sampling of 47 randomly selected Thoroughbreds were endoscopically examined for EIPH...
Clinical effects and pharmacokinetics of nebulized lidocaine in healthy horses. Nebulized lidocaine appears promising as a novel corticosteroid-sparing therapeutic for equine asthma, but its safety and pharmacokinetic behavior have yet to be confirmed. Unassigned: To describe the effect of nebulized lidocaine on upper airway sensitivity, lung mechanics, and lower respiratory cellular response of healthy horses, as well as delivery of lidocaine to lower airways, and its subsequent absorption, clearance, and duration of detectability. Unassigned: Six healthy university- and client-owned horses with normal physical examination and serum amyloid A, and no history of respirato...
Single-cell gene expression analysis of cryopreserved equine bronchoalveolar cells. The transcriptomic profile of a cell population can now be studied at the cellular level using single-cell mRNA sequencing (scRNA-seq). This novel technique provides the unprecedented opportunity to explore the cellular composition of the bronchoalveolar lavage fluid (BALF) of the horse, a species for which cell type markers are poorly described. Here, scRNA-seq technology was applied to cryopreserved equine BALF cells. Analysis of 4,631 cells isolated from three asthmatic horses in remission identified 16 cell clusters belonging to six major cell types: monocytes/macrophages, T cells, B/plasm...
Neutrophil Extracellular Traps Are Found in Bronchoalveolar Lavage Fluids of Horses With Severe Asthma and Correlate With Asthma Severity. Asthma encompasses a spectrum of heterogenous immune-mediated respiratory disorders sharing a similar clinical pattern characterized by cough, wheeze and exercise intolerance. In horses, equine asthma can be subdivided into severe or moderate asthma according to clinical symptoms and the extent of airway neutrophilic inflammation. While severe asthmatic horses are characterized by an elevated neutrophilic inflammation of the lower airways, cough, dyspnea at rest and high mucus secretion, horses with moderate asthma show a milder neutrophilic inflammation, exhibit intolerance to exercise but no...
Lipidomic analysis of surfactant and plasma from horses with asthma and age-matched healthy horses. To perform lipidomic analysis of surfactant and plasma from asthmatic and healthy horses. Methods: 30 horses with clinical signs of asthma and 30 age-matched control horses. Methods: Detailed history, physical examination, CBC, and bronchoalveolar lavage fluid (BALF) cytologies were obtained. Asthmatic horses were grouped based on their BALF inflammatory profile: severe equine asthma (SEA), mild equine asthma with neutrophilic airway inflammation (MEA-N), or mild equine asthma with eosinophilic airway inflammation (MEA-E). Each asthma group was assigned its own age-matched control group. Lipid...
Mild-moderate equine asthma: A scoping review of evidence supporting the consensus definition. Current consensus defines mild-moderate equine asthma (mEA; previously inflammatory airway disease) by a hierarchy of indicators of lung pathology: cough, poor performance, increased tracheobronchial mucus, inflammatory bronchoalveolar lavage (BAL) cytology and pulmonary dysfunction. Exclusion criteria include fever, systemic disease, or increased resting respiratory effort. The aim of this review was to inform future research by identifying gaps, strengths and weaknesses in the current body of evidence supporting this consensus-proposed definition. Objectives were to critique evidence support...
Comprehensive Flow Cytometric Characterization of Bronchoalveolar Lavage Cells Indicates Comparable Phenotypes Between Asthmatic and Healthy Horses But Functional Lymphocyte Differences. Equine asthma (EA) is a highly relevant disease, estimated to affect up to 20% of all horses, and compares to human asthma. The pathogenesis of EA is most likely immune-mediated, yet incompletely understood. To study the immune response in the affected lower airways, mixed leukocytes were acquired through bronchoalveolar lavage (BAL) and the cell populations were analyzed on a single-cell basis by flow cytometry (FC). Samples of 38 horses grouped as respiratory healthy or affected by mild to moderate (mEA) or severe EA (sEA) according to their history, clinical signs, and BAL cytology were ana...
Isolation of Extracellular Vesicles From the Bronchoalveolar Lavage Fluid of Healthy and Asthmatic Horses. Extracellular vesicles (EVs) are membrane-bound particles that engage in inflammatory reactions by mediating cell-cell interactions. Previously, EVs have been isolated from the bronchoalveolar lavage fluid (BALF) of humans and rodents. The aim of this study was to investigate the number and size distribution of EVs in the BALF of asthmatic horses (EA, = 35) and healthy horses ( = 19). Saline was injected during bronchoscopy to the right lung followed by manual aspiration. The retrieved BALF was centrifuged twice to remove cells and biological debris. The supernatant was concentrated and EVs w...
Upper and Lower Airways Evaluation and Its Relationship with Dynamic Upper Airway Obstruction in Racehorses. Dynamic upper airway obstructions (DUAO) are common in racehorses, but their pathogenetic mechanisms have not been completely clarified yet. Multiple studies suggest that alterations of the pharyngo-laryngeal region visible at resting endoscopy may be predictive of the onset of DUAO, and the development of DUAO may be associated with pharyngeal lymphoid hyperplasia (PLH), lower airway inflammation (LAI) and exercise-induced pulmonary hemorrhage (EIPH). The present study aims to investigate the possible relationship between the findings of a complete resting evaluation of the upper and lower ai...
Association between dynamic upper airway obstructions and fitness parameters in Standardbred racehorses during high-speed treadmill exercise. To evaluate the association between the severity of dynamic upper airway obstruction (DUAO) and selected fitness parameters in Standardbred racehorses measured by treadmill test. 191 Standardbred racehorses that underwent a standardized diagnostic protocol for poor performance evaluation between 2002 and 2021 at the Veterinary Teaching Hospital, University of Milan. Horses underwent a diagnostic protocol including clinical examination, lameness evaluation, laboratory analyses, ECG, resting upper airway endoscopy, metabolic incremental treadmill test, high-speed treadmill endoscopy, postexercis...
Characterization of fungal exposure and dectin-1 expression in healthy horses and horses with severe asthma. To quantify dectin-1 expression in bronchoalveolar lavage fluid (BALF), create polyclonal antibodies against equine dectin-1 and localize it in tissues, and quantify fungal exposure in pastured and stabled asthmatic and nonasthmatic horses. Methods: BALF samples from 6 controls and 6 horses with severe asthma. Stored lung and nasal wash samples. Methods: Dectin-1 expression was quantified by quantitative PCR (qPCR). Purified peptide from equine dectin-1 was used to generate polyclonal antibodies and was confirmed with immunological testing. Fungal exposure was quantified in BALF samples by cou...
Giant Multinucleated Cells Are Associated with Mastocytic Inflammatory Signature Equine Asthma. Equine asthma is currently diagnosed by the presence of increased neutrophil (>5%), mast cell (>2%), and/or eosinophil (>1%) differential cell count. Macrophages are normal resident cells within the alveoli. Their presence in BALF is considered normal, but the clinical implication of the presence of activated or fused macrophages (giant multinucleated cells, GMC) is currently overlooked. We aimed to assess the prevalence, cytological determinants, and clinical significance of increased GMC counts in BALF of 34 asthmatic horses compared to 10 controls. Counts were performed on 15 rando...
Associations between Exercise-Induced Pulmonary Hemorrhage (EIPH) and Fitness Parameters Measured by Incremental Treadmill Test in Standardbred Racehorses. Exercise-induced pulmonary hemorrhage (EIPH) is a condition affecting up to 95% of racehorses, diagnosed by detecting blood in the trachea after exercise and/or the presence of hemosiderophages in the bronchoalveolar lavage fluid (BALf). Although EIPH is commonly associated with poor performance, scientific evidence is scarce. The athletic capacity of racehorses can be quantified through some parameters obtained during an incremental treadmill test; in particular, the speed at a heart rate of 200 bpm (V200), and the speed (VLa4) and the heart rate (HRLa4) at which the blood lactate concentrati...
Effect of intrabronchial administration of autologous adipose-derived mesenchymal stem cells on severe equine asthma. Severe equine asthma (SEA) is a common chronic respiratory disease and a significant health and well-being problem in horses. Current therapeutic strategies improve pulmonary function and clinical signs in some horses, but in the long-term, return to full athletic function appears to be rare. The aim of this study was to assess the safety and the effect of intrabronchial administration of adipose-derived mesenchymal stem cells (AD-MSC) on pulmonary inflammatory and clinical parameters in horses with SEA. This was a randomized controlled trial. Twenty adult horses diagnosed with SEA were random...
Effects of Bedding Material on Equine Lower Airway Inflammation: A Comparison of Two Peat Beddings, Wood Pellet, and Straw Pellet. The effects of bedding material on air quality are important amongst horses worldwide. Respiratory diseases, especially equine asthma, are highly prevalent with air hygiene playing a major role on the pathophysiology of these diseases. The objective of our study was to investigate the effects of four bedding materials on the respiratory signs, tracheal mucus score, and tracheal wash (TW) and bronchoalveolar lavage fluid (BALF) cytology in healthy adult horses. The study design was a prospective controlled cross-over study, and the subjects were healthy adult riding school horses ( = 32) from a...
Bronchial brush cytology, endobronchial biopsy, and SALSA immunohistochemistry in severe equine asthma. Horses with severe equine asthma (SEA), also known as heaves and recurrent airway obstruction, have persistent neutrophilic inflammation of the lower airways. Cytologic evaluation of bronchoalveolar lavage (BAL) fluid is commonly used to confirm the clinical diagnosis of SEA. However, the utility of microscopic assessment of bronchial brushings, endobronchial biopsies, and immunohistochemical detection of disease-associated biomarkers for the diagnosis of SEA remain poorly characterized. Salivary scavenger and agglutinin (SALSA) has anti-inflammatory properties and downregulated gene expressio...
Airway remodeling in horses with mild and moderate asthma. There is a remodeling of the central airways in horses with severe asthma but whether a similar process occurs in horses with the mild or moderate asthma (MMA) is unknown. Objective: To evaluate lesions affecting the central airways of horses with MMA. Methods: Twelve horses with MMA and 8 control horses. Methods: Case-control retrospective study of horses classified as MMA affected or controls based on history and bronchoalveolar lavage fluid cytology. Endobronchial biopsies were analyzed using histomorphometry and a semiquantitative histologic scoring system. Results: Histomorphometry identi...
Effects of forages, dust exposure and proresolving lipids on airway inflammation in horses. To investigate the role of omega-3 polyunsaturated fatty acids (Ω-3)-derived proresolving lipid mediators (PRLM) in the resolution of mild airway inflammation in horses. Methods: 20 horses with mild airway inflammation. Methods: Horses previously eating hay were fed hay pellets (low Ω-3 content; n = 10) or haylage (high Ω-3 content; 9) for 6 weeks. Dust exposure was measured in the breathing zone with a real-time particulate monitor. Bronchoalveolar lavage (BAL) was performed at baseline, week 3, and week 6. The effect of PRLM on neutrophil apoptosis and efferocytosis was examined in vitro....
The Role of Thoracic Ultrasonography and Airway Endoscopy in the Diagnosis of Equine Asthma and Exercise-Induced Pulmonary Hemorrhage. Mild-moderate (MEA), severe (SEA) equine asthma and exercise-induced pulmonary hemorrhage (EIPH) are common respiratory disorders in horses. The present retrospective study aims to evaluate the role of ultrasonography and endoscopy in the diagnosis of these conditions. Three hundred and three horses were included and divided into SEA, MEA and MEA + EIPH groups, on the basis of history, clinical examination and bronchoalveolar lavage fluid (BALf) cytology; scores were assigned to lung ultrasonography, pharyngeal lymphoid hyperplasia (PLH), tracheal mucus (TM) and tracheal bifurcation edema (TB)...
Flow cytometric analysis of equine bronchoalveolar lavage fluid cells in horses with and without severe equine asthma. Severe equine asthma (SEA) is a common, debilitating lower airway inflammatory disorder of older horses. Alveolar macrophages (AMs) survey inhaled particulates from barn sources causing them to switch from an anti-inflammatory to a proinflammatory phenotype, resulting in neutrophil recruitment to the lung. This proinflammatory switch may contribute to the development and prolongation of SEA. Validated antibodies to identify the cells involved in the pathogenesis of SEA are lacking. In this study, monoclonal antibodies against CD90, CD163, and CD206 were tested for reactivity with equine leukoc...
The effect of lower airway inflammation on inflammatory cytokine gene expression in bronchoalveolar lavage fluid and whole blood in racing Thoroughbreds. Immunological mechanisms involved in the pathogenesis of mild to moderate equine asthma (MEA) are not completely understood. There are limited data on bronchoalveolar lavage fluid (BALF) and blood inflammatory cytokine profiles in racehorses with MEA, and the effect of racing on inflammatory cytokines is unknown. Objective: We hypothesized that inflammatory cytokine gene expression in BALF and resting blood would be higher in racehorses with lower airway inflammation compared to healthy controls, and that gene expression in blood collected immediately post-race would be increased compared to r...