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.
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...
Nicoletella semolina in the airways of healthy horses and horses with severe asthma. Nicoletella semolina was identified in the airways of horses and its low prevalence could be because of its difficult differentiation from other Pasteurellaceae. Objective: To develop a molecular method for the identification of N. semolina and to evaluate its prevalence in the mouth and the airways of healthy and severe asthmatic horses. Methods: Six healthy and 6 severely asthmatic horses in phase I, 10 severely asthmatic horses in phase II, and 10 healthy horses in phase III. Methods: Cohort (phases I and II) and cross-sectional (phase III) studies. Quantitative polymerase chain reaction pr...
Equine respiratory viruses, airway inflammation and performance in thoroughbred racehorses. Equine asthma is a common cause of poor performance in racehorses but it is unclear if respiratory viruses contribute to its etiology. The objective of the study was to determine if respiratory viruses were associated with clinical signs and bronchoalveolar lavage fluid (BALF) cytology in Thoroughbred racehorses. Equine herpesviruses (EHV-1, 2, 4, 5) and equine rhinitis A and B viruses (ERBV, ERAV) genomes were quantified by qPCR in nasopharyngeal, tracheal, and BALF samples collected after racing. The relationships between virus detection and load and clinical signs, performance, BALF cytolog...
Effects of Bedding Material on Equine Lower Airway Inflammation: A Crossover Study Comparing Peat and Wood Shavings. Bedding materials affect stable air hygiene, and thus the development and exacerbation of equine asthma. There is limited knowledge concerning the effects of different types of bedding material on equine lower airway inflammation. The objective of our study was to investigate the effects of bedding materials on respiratory signs, tracheal mucus score, and lower airway cytology in healthy adult horses. The study design was a prospective controlled study, and the subjects were healthy adult riding school horses (n = 32) from a single stable. Wood shavings were compared to peat, which was used as...
Pharmacokinetics and pulmonary distribution of Draxxin® (tulathromycin) in healthy adult horses. The objective of this study was to determine the pharmacokinetics and tolerability of tulathromycin (Draxxin ; 2.5 mg/kg once) after intramuscular (IM), subcutaneous (SC), and slow intravenous (IV) administration to six adult horses. A three-phase design and 4-week washout period were used. Drug concentrations in blood and bronchoalveolar lavage (BAL) samples were determined by ultra-performance liquid chromatography tandem mass spectrometry and pharmacokinetic parameters calculated using noncompartmental analysis. Following SC and IM administration, all horses exhibited sweating, discomfort,...
Evaluation of Treatment With Respiratory Gene Technology and Serum in a Group of Standard Bred Racehorses With Cytological Evidence of Mild Equine Asthma. Alternative treatment options to glucocorticoids for equine asthma is desirable due to withdrawal time. The objective was to evaluate if serum and Respiratory Gene Technology (RGT), a commercial kit to produce autologous conditioned serum, was effective in reducing bronchoalveolar lavage (BAL) neutrophils and mast cells in racehorses with cytological evidence of mild equine asthma . Thirty-six Standardbred trotters in active training were enrolled in this randomized clinical trial; a healthy control group (n=11), a RGT group (n=12) and a serum group (n=13). Endoscopy including tracheal wash (T...
Bronchoalveolar lavage fluid cytological findings in healthy Amiata donkeys. The approach to respiratory diseases in donkeys is similar to that for horses; nevertheless, Bronchoalveolar lavage fluid (BALF) and tracheal wash cytology in this species have been described only a few times in the literature. To describe BALF cytological findings in a cohort of 24 healthy Amiata donkeys. Bronchoalveolar lavage (BAL) was carried out via standing sedation with a large animal where the BAL catheter passed blindly through the nasal passage into the trachea. The total nucleated cell count of the BALF was found similar to that already described in healthy horses and donkeys. No di...
Differential gene expression and Ingenuity Pathway Analysis of bronchoalveolar lavage cells from horses with mild/moderate neutrophilic or mastocytic inflammation on BAL cytology. Mild to moderate equine asthma syndrome (mEAS) affects horses of all ages and breeds. To date, the etiology and pathophysiology of mEAS are active areas of research, and it remains incompletely understood whether mEAS horses with different immune cell 'signatures' on BAL cytology represent different phenotypes, distinct pathobiological mechanisms (endotypes), varied environmental conditions, disease severity, genetic predispositions, or all of the above. In this descriptive study, we compared gene expression data from BAL cells isolated from horses with normal BALF cytology (n = 5), to those i...
Surfactant protein D concentrations in serum and bronchoalveolar lavage fluid from young healthy horses on pasture and in a barn environment. To evaluate surfactant protein D (SP-D) concentrations in serum and bronchoalveolar lavage fluid (BALF) from young healthy horses on pasture or housed in a typical barn. Methods: 20 young healthy horses. Methods: Horses were randomly assigned to 1 of 2 groups (pasture, n = 10; barn, 10), and serum and BALF samples were collected for SP-D determination at baseline (all horses on pasture) and 2 weeks and 4 weeks after the barn group of horses was relocated from the pasture to the barn. Other evaluations included physical and tracheoscopic examinations. Findings were compared within and between g...
Opsonization but not pretreatment of equine macrophages with hyperimmune plasma nonspecifically enhances phagocytosis and intracellular killing of Rhodococcus equi. Evidence regarding the efficacy of equine hyperimmune plasma to prevent pneumonia in foals caused by Rhodococcus equi is limited and conflicting. Objective: Opsonization with R. equi-specific hyperimmune plasma (HIP) will significantly increase phagocytosis and decrease intracellular replication of R. equi by alveolar macrophages (AMs) compared to normal plasma (NP). Methods: Fifteen adult Quarter Horses were used to collect bronchoalveolar lavage cells. Methods: In the first experiment, AMs from 9 horses were pretreated (incubated) with either HIP, NP, or media only (control) and then infecte...
Asthmatic Disease as an Underlying Cause of Dorsal Displacement of the Soft Palate in Horses. It is important to treat the upper and lower respiratory tracts as a single unit, as lower respiratory tract diseases can often cause upper respiratory functional disorders, whereas upper respiratory obstructions could be a factor in lower respiratory problems. The present study aimed to investigate the hypothesis that asthmatic diseases may be an underlying cause of dorsal displacement of the soft palate in horses. Pleasure or sport horses (n = 57) with a history of asthmatic disease were incorporated in the study. All horses were examined in the exacerbation phase of the asthmatic disease. ...
Validation of a Rapid Bronchoalveolar Lavage Fluid Colorimetric Evaluation for Assessing the Severity of Exercise-Induced Pulmonary Hemorrhage in Horses in Field Conditions. Diagnosis and assessment of severity of exercise-induced pulmonary hemorrhage (EIPH) relies on postexercise visualization of fresh blood in the airways via tracheobronchoscopic examination (TBE) and/or counting erythrocytes in bronchoalveolar lavage fluid (BALFRBC). Determining the BALFRBC is more sensitive than TBE but its usefulness is hampered by the need to have BALFRBC counted at a laboratory. We explored the feasibility of evaluating the severity of EIPH by using a color chart comprised of five shades of red and matching those colors with the color of BALF immediately following collectio...
Metabolomics analysis of bronchoalveolar lavage fluid samples in horses with naturally-occurring asthma and experimentally-induced airway inflammation. The present work characterized the metabolomic profile of bronchoalveolar lavage fluid (BALF) in healthy horses, experimentally-induced airway inflammation by lipopolysaccharide (LPS) nebulization, and naturally-occurring asthma (n = 3 in each group). All animals underwent clinical and upper airway endoscopic examinations, and bronchoalveolar lavage. BALF supernatant samples were subjected to metabolic analysis based on gas chromatography-mass spectrometry (GC-MS). Overall, 67 peaks were obtained from BALF GC-MS analysis, corresponding to 53 metabolites which were categorized according to ch...
Clinical Feasibility and Airway Deposition of Nebulized Voriconazole in Healthy Horses. Voriconazole (VRC) is a potential treatment for pneumomycosis in horses. The objectives of this study were to determine if the delivery of Vfend using a Flexineb nebulizer produced clinically significant [VRC] in lower airways. The hypothesis was that [VRC] after delivery by nebulization would be greater in the pulmonary epithelial lining fluid than plasma. A secondary objective was to determine [VRC] in upper airways through the collection of nasopharyngeal wash (NPW) samples. Voriconazole solution [Vfend-6.25 mg/mL, 100 (n = 2), 200 (n = 3), 500 (n = 1) mg] was nebulized once in 6 health...
Deep Learning-Based Quantification of Pulmonary Hemosiderophages in Cytology Slides. Exercise-induced pulmonary hemorrhage (EIPH) is a common condition in sport horses with negative impact on performance. Cytology of bronchoalveolar lavage fluid by use of a scoring system is considered the most sensitive diagnostic method. Macrophages are classified depending on the degree of cytoplasmic hemosiderin content. The current gold standard is manual grading, which is however monotonous and time-consuming. We evaluated state-of-the-art deep learning-based methods for single cell macrophage classification and compared them against the performance of nine cytology experts and evaluated...
The Relationship between Lung Inflammation and Aerobic Threshold in Standardbred Racehorses with Mild-Moderate Equine Asthma. Mild-moderate equine asthma (MEA) is a common disease of young racehorses characterized by inflammation of the lower airways diagnosed by cytological examination of bronchoalveolar lavage fluid (BALF), and it is considered an important cause of poor performance. The most common parameter of athletic capacity associated with horse performance is the speed at 4 mmol/L of lactate (VLA4). The aim of the present work was to evaluate the relation between the different BALF inflammatory cells and VLA4 in racehorses affected by MEA. A population of Standardbred racehorses that underwent an accurate pr...
A retrospective study of bronchoalveolar lavage fluid analysis in barrel racing horses with exercise-induced pulmonary hemorrhage and asthma in Texas from 2016 to 2018. Exercise-induced pulmonary hemorrhage (EIPH) and asthma in barrel racing horses is a common disease across the United States. Limited information is available on non-infectious respiratory diseases in this population, the interaction between these two diseases, and the occurrence of both EIPH and asthma in the horse. The purpose of this study was to evaluate the bronchoalveolar lavage (BAL) fluid cytological results of barrel racing horses with EIPH, asthma, or both. A retrospective study was conducted using the medical records of horses that presented with cough and decreased athletic perform...
Respiratory metabolites in bronchoalveolar lavage fluid (BALF) and exhaled breath condensate (EBC) can differentiate horses affected by severe equine asthma from healthy horses. The use of an untargeted metabolomic approach to investigate biofluids of respiratory origin is of increasing interest in human and veterinary lung research. Considering the high incidence of equine asthma (> 14%) within horse population and the importance of this animal model for human disease, we aimed to investigate the metabolomic profile of bronchoalveolar lavage fluid (BALF) and exhaled breath condensate (EBC) in healthy and asthmatic horses. Results: On the basis of clinical, endoscopic and BALF cytology findings, 6 horses with severe asthma (Group A) and 6 healthy horses (Group C) we...
Physiologic effects of furosemide in combination with water restriction when administered at 4 and 24 hours prior to high-intensity treadmill training. Although controversial, due to its reported effectiveness in attenuating bleeding associated with exercise-induced pulmonary hemorrhage (EIPH), furosemide is currently a permitted race day medication in most North American racing jurisdictions. The objective of this study was to assess the efficacy of furosemide in reducing the presence and severity of EIPH when administered 24 hr prior to strenuous treadmill exercise. Eight exercised Thoroughbred horses received saline or 250 mg of furosemide either 4 or 24 hr prior to high-speed treadmill exercise in a balanced 3-way cross-over design. Bl...