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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.
A CONSORT-guided, randomized controlled clinical trial of nebulized administration of dexamethasone and saline on lower airway cytokine mRNA expression in horses with moderate asthma. Nebulized administration of dexamethasone on cytokine regulation in horses with moderate asthma has not been investigated. Objective: To investigate the changes in expression of inflammatory cytokine mRNA after nebulized administration of dexamethasone treatment of horses with moderate asthma. Methods: Horses with naturally occurring moderate asthma (n = 16) and healthy control horses (n = 4). All horses were kept in a dusty environment during the study. Methods: Prospective, parallel, randomized, controlled, blinded clinical trial. Blood endogenous cortisol, tracheal mucus, and bronch...
The impact of age on vitamin D receptor expression, vitamin D metabolism and cytokine production in ex vivo Rhodococcus equi infection of equine alveolar macrophages. Rhodococcus equi (R. equi), a pneumonia-causing intracellular bacterium, results in significant morbidity and mortality in young foals, while healthy adult horses rarely develop disease. Survival and replication within alveolar macrophages (AMφ) are the hallmarks of R. equi's pathogenicity. The vitamin D receptor (VDR) and its ligand, the active vitamin D metabolite 1,25(OH)2D, are important in immune responses to intracellular bacteria. The vitamin D/VDR pathway regulates the downstream production of cytokines in infected human AMφ. The immunomodulatory role of the vitamin D/VDR pathway in ...
Intramuscular but not nebulized administration of a mRNA vaccine against Rhodococcus equi stimulated humoral immune responses in neonatal foals. Design and evaluate immune responses of neonatal foals to a mRNA vaccine expressing the virulence-associated protein A (VapA) of Rhodococcus equi. Methods: Cultured primary equine respiratory tract cells; Serum, bronchoalveolar lavage fluid (BALF), and peripheral blood mononuclear cells (PBMCs) from 30 healthy Quarter Horse foals. Methods: VapA expression was evaluated by western immunoblot in cultured equine bronchial cells transfected with 4 mRNA constructs encoding VapA. The mRNA construct with greatest expression was used to immunize foals at ages 2 and 21 days in 5 groups: (1) 300 μg neb...
Lipid species profiling of bronchoalveolar lavage fluid cells of horses housed on two different bedding materials. The lipidome of equine BALF cells has not been described. The objectives of this prospective repeated-measures study were to explore the BALF cells' lipidome in horses and to identify lipids associated with progression or resolution of airway inflammation. BALF cells from 22 horses exposed to two bedding materials (Peat 1-Wood shavings [WS]-Peat 2) were studied by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The effects of bedding on lipid class and species compositions were tested with rmANOVA. Correlations between lipids and cell counts were examined. The BALF cells' lipidome s...
A Longitudinal Analysis of Equine Asthma Presentation and Response to Treatment Using Lung Function Testing and BAL Cytology Analysis in Combination with Owner Perception. (1) Background: Equine asthma (EA) is a pervasive and important cause of poor performance and respiratory morbidity in horses. Diagnosis of EA includes an owner complaint, clinical scoring, lung function testing, and cytological analysis of bronchoalveolar lavage (BAL) cytology. There is a paucity of information about the longitudinal course of the disease using these outcome assessments; thus, this study sought to describe and quantify, in horses with more than one visit to a specialty pulmonary clinic in New England, the type and range of clinical presentations with an eventual diagnosis of ...
Single-cell transcriptomics delineates the immune cell landscape in equine lower airways and reveals upregulation of FKBP5 in horses with asthma. Equine asthma (EA) is a heterogenous, complex disease, with a significant negative impact on horse welfare and performance. EA and human asthma share fundamental similarities, making EA a useful model for studying the disease. One relevant sample type for investigating chronic lung inflammation is bronchoalveolar lavage fluid (BALF), which provides a snapshot of the immune cells present in the alveolar space. To investigate the immune cell landscape of the respiratory tract in horses with mild-to-moderate equine asthma (mEA) and healthy controls, single-cell RNA sequencing was conducted on equ...
Molecular and Cellular Evaluation of Horses With Summer Pasture Associated Asthma Syndrome. Equine asthma is an airway disease that affects a large number of horses annually leading to considerable economic losses in the horse industry. Despite advances in research in this area, there is still a lack of information on its etiology and molecular characterization in pasture associated asthma. The objective of the current study was to characterize the inflammatory disease of lower airways in horses maintained on pasture through cytologic and immunologic profile during the summer in a tropical environment by analysis of the gene expression of Th1 cytokines (IFN- λ, IL-8), Th2 cytokines ...
Detection of fungi in the airways of horses according to the sample site: a methodological study. Fungal detection in equine airways may be performed on either tracheal wash (TW) or bronchoalveolar lavage fluid (BALF) by either cytology or culture. However, method comparisons are sparse. Our objective was to determine the prevalence of fungi in airways of horses according to the sample site and laboratory methodology. Sixty-two adult horses, investigated in the field or referred for respiratory disease, were included. Tracheal wash, and BALF collected separately from both lungs, were collected using a videoendoscope. Fungi were detected in cytologic samples examined by light microscopy, an...
Comparison of the broncoalveolar lavage fluid proteomics between foals and adult horses. Neonates have different cellular composition in their bronchoalveolar lavage fluid (BALF) when compared to foals and adult horses; however, little is known about the non-cellular components of BALF. The objective of this study was to determine the proteomic composition of BALF in neonatal horses and to compare it to that of foals and adult horses. Bronchoalveolar lavage fluid samples of seven neonates (< 1 week age), four 5 to 7-week-old foals, and six adult horses were collected. Quantitative proteomics of the fluid was performed using tandem mass tag labeling followed by high resolution l...
Atrial fibrillation as a risk factor for exercise-induced pulmonary haemorrhage following a standardised exercise test. Atrial fibrillation (AF) has been proposed as a risk factor for exercise-induced pulmonary haemorrhage (EIPH) due to increased pressure in the left atrium. Objective: To evaluate if AF was associated with EIPH following a standardised exercise test (SET) to fatigue. Methods: Two-arm controlled experiment. Methods: Ten untrained Standardbred mares mean (standard deviation [SD]) age 6 (2) years performed a SET on the treadmill in sinus rhythm (SR) (SET1) and 25-44 days after induction of self-sustained AF (SET2). AF was induced by tachypacing using a pacing device. Endoscopy, including trachea...
Bronchoalveolar Lavage Cytology in Severe Equine Asthma: Cytocentrifugated versus Sediment Smear Preparations. Equine asthma is a common respiratory disease that may affect horses of any age. The diagnosis of severe equine asthma (SEA) (historically referred as recurrent airway obstruction or RAO) is based mainly on the history of the animal and clinical signs, which are further supported by the cytological examination of the bronchoalveolar lavage (BAL). This can also be helpful in monitoring the inflammation of the lower airways in response to environmental management and medication. The cytocentrifugated preparation is usually considered the method of choice for BAL cytological interpretation. The a...
Effect of Lavage Solution Type on Bronchoalveolar Lavage Fluid Cytology in Clinically Healthy Horses. Equine bronchoalveolar lavage (BAL) is usually performed with 250-500 mL of isotonic saline at pH 5.5. The acidic pH of saline may cause an increase in airway neutrophil count 48 h after BAL. Other isotonic solutions such as Ringer's solution, phosphate-buffered saline (PBS) or Plasma-Lyte 148 have a neutral pH of 7.4 and might be a better choice for BAL by not provoking inflammation and the influx of neutrophils into airways. BAL was performed in four healthy horses in four different lung lobes using four different solutions in a randomized crossover design. In each lobe, BAL was performed tw...
Airway Hyperresponsiveness, but Not Bronchoalveolar Inflammatory Cytokines Profiles, Is Modified at the Subclinical Onset of Severe Equine Asthma. Airway hyperresponsiveness (AHR) and inflammation are both observed in human and equine asthma. The aim of this study was to assess the timeline and relationship of both features at the subclinical onset of severe equine asthma (SEA). First, the repeatability of the pulmonary function test (PFT) using impulse oscillometry system, and the methacholine bronchoprovocation test (BPT) were assessed at a 1-day interval on six SEA horses in clinical remission and six control horses. Then, clinical and ancillary tests were performed before and after a 1-week low-dust environmental challenge, including...
Fatty acid fingerprints in bronchoalveolar lavage fluid and its extracellular vesicles reflect equine asthma severity. Equine asthma (EA) is an inflammatory disease of the lower airways driven by mediators released from cells. Extracellular vesicles (EVs) are vehicles for lipid mediators, which possess either pro-inflammatory or dual anti-inflammatory and pro-resolving functions. In this study, we investigated how the respiratory fatty acid (FA) profile reflects airway inflammatory status. The FA composition of bronchoalveolar lavage fluid (BALF), BALF supernatant, and bronchoalveolar EVs of healthy horses (n = 15) and horses with mild/moderate EA (n = 10) or severe EA (SEA, n = 5) was determined w...
Medical causes of poor performance and their associations with fitness in Standardbred racehorses. Poor performance is a multifactorial syndrome of racehorses, commonly associated with subclinical disorders, which can be diagnosed by exercise testing. Objective: Describe the prevalence of medical causes of poor performance in Standardbreds unassociated with lameness, and evaluate their relationships with fitness variables measured by exercise treadmill test. Methods: Hospital population of 259 nonlame Standardbred trotters referred for poor performance. Methods: The horses' medical records were retrospectively reviewed. Horses underwent a diagnostic protocol including resting examination, p...
Neutrophil gelatinase-associated lipocalin as a potential biomarker for equine asthma. Studies 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). Objective: To 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). Methods: Retrospective cross-sectional study. Methods: Details of endoscopic examination including tracheal mucus score (TMS, scale 0-5) and BAL cytolog...
Bronchoalveolar lavage hemosiderosis in lightly active or sedentary horses. Hemosiderophages in bronchoalveolar lavage fluid (BALF) are commonly ascribed to exercise-induced pulmonary hemorrhage (EIPH). Little information exists regarding the presence of these cells in horses that perform light or no work and that are referred for respiratory problems. Objective: Evaluate the presence of hemosiderophages in BALF of horses suspected of respiratory disease without history of or risk factors for EIPH and determine predictors of hemosiderophages in BALF in this population. Methods: Observational retrospective cross-sectional study using STROBE (Strengthening the Reporting...
Erratum: Single-cell gene expression analysis of cryopreserved equine bronchoalveolar cells. [This corrects the article DOI: 10.3389/fimmu.2022.929922.].
Diagnosis of Aspergillosis in Horses. Invasive pulmonary aspergillosis (IPA) may be a rare cause of granulomatous pneumonia in horses. The mortality of IPA is almost 100%; direct diagnostic tools in horses are needed. Bronchoalveolar lavage fluid (BALF) and serum samples were collected from 18 horses, including individuals suffering from IPA (n = 1), equine asthma (EA, n = 12), and 5 healthy controls. Serum samples were collected from another 6 healthy controls. Samples of BALF (n = 18) were analyzed for spp. DNA, fungal galactomannan (GM), ferricrocin (Fc), triacetylfusarinin C (TafC), and gliotoxin (Gtx). Analysis of 24 serum s...
Initial investigation of molecular phenotypes of airway mast cells and cytokine profiles in equine asthma. Equine asthma is a naturally occurring lung disease characterized by chronic, partially reversible airway obstruction, pulmonary remodeling, and lower airway inflammation. Asthma is currently divided into two major groups, mild to moderate asthma (mEA) and severe asthma (sEA), but further subtyping by phenotype (i.e., clinical presentation) and/or endotype (i.e., cellular mechanisms) may be warranted. For this study, we were interested in further investigation of cellular and inflammatory characteristics of EA, including airway mast cells. The purpose of this study was to: (1) compare mast cel...
Gene Expression Profiles of the Immuno-Transcriptome in Equine Asthma. Mild equine asthma (MEA) and severe equine asthma (SEA) are two of the most frequent equine airway inflammatory diseases, but knowledge about their pathogenesis is limited. The goal of this study was to investigate gene expression differences in the respiratory tract of MEA- and SEA-affected horses and their relationship with clinical signs. Methods: Clinical examination and endoscopy were performed in 8 SEA- and 10 MEA-affected horses and 7 healthy controls. Cytological and microbiological analyses of bronchoalveolar lavage (BAL) fluid were performed. Gene expression profiling of BAL fluid wa...
Protein microarray allergen profiling in bronchoalveolar lavage fluid and serum of horses with asthma. The diagnostic value of allergen-specific immunoglobulin E (IgE) in horses with asthma is uncertain. A recently developed protein microarray detected abnormally high latex-specific IgE concentrations in the serum of horses with severe asthma. Objective: The main objective was to characterize the IgE profiles of asthmatic horses in Switzerland using a protein microarray platform in serum and bronchoalveolar lavage fluid (BALF). The secondary objective was to determine whether serological and BALF allergen-specific IgE concentrations correlated. Methods: Forty-four asthmatic and 39 control horse...
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...
