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 (n = 32) from a single stable. Wood pellet, straw pellet, and loosely stored peat (Peat 3) were compared to peat packed in plastic-covered bales (Peat 2). Lower airway endoscopy and sampling (TW and BALF) for cytological examination were performed after each 35-day bedding period. The tracheal mucus scores (P = 0.014) and respiratory rate (P = 0.026) were higher during the straw pellet period compared to the Peat 2 period. The respiratory rate was lower during the wood pellet period compared to the Peat 2 period (P = 0.004). The TW neutrophil percentage during the straw pellet period was higher compared to the Peat 2 period (P = 0.0003). The BALF neutrophil percentage was higher during the straw pellet period (P = 0.005) and during the Peat 3 period compared to the Peat 2 period (P = 0.04). We conclude that baled peat (Peat 2) caused lower neutrophil percentages in the airway samples compared to straw pellet and loosely stored peat (Peat 3). No difference was observed between Peat 2 and wood pellet. The information gained from this study may assist veterinarians and horse owners in selecting appropriate bedding materials, especially for horses with equine asthma.

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 expression in SEA; therefore, it was investigated as a tissue biomarker for airway and systemic inflammation. Six asthmatic and 6 non-asthmatic horses were exposed to an inhaled challenge. Before and after challenge, samples of BAL fluid, bronchial brushing, and endobronchial biopsy were collected. Location of SALSA in biopsies was determined, and immunohistochemical label intensity was computed using image analysis software. Serum amyloid A (SAA) was measured to assess systemic inflammation. After challenge, neutrophil proportions were significantly higher in asthmatic versus non-asthmatic horses in BAL fluid (least squares means, 95% confidence interval: 80.9%, 57.2% to 93.1%, vs 3.6%, 1.1% to 10.7%) and in brush cytology slides (39.5%, 7.7% to 83.6%, vs 0.2%, 0% to 2.3%), illustrating the potential of brush cytology as an alternate modality to BAL for assessing intraluminal inflammation. Bronchial histopathologic findings and intensity of SALSA immunolabeling in surface and glandular epithelium were similar in asthmatic and non-asthmatic horses, indicating limited changes in bronchial tissue from the inhaled challenge. Increases in SAA indicated systemic inflammation, but SALSA immunolabeling did not change significantly.

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). These scores were compared between groups, and their associations with age, BALf cytology, tracheal wash microbiology and between endoscopic and ultrasonographic scores were statistically analyzed. Ultrasonographic scores were higher in the SEA and MEA + EIPH groups and associated with increased BALf neutrophils and hemosiderophages. The PLH score was higher in younger horses affected by MEA and EIPH and associated with increased eosinophils and hemosiderophages. TM and TB scores were greater in older horses affected by SEA, associated with increased neutrophils and inversely correlated with hemosiderophages. Moreover, TM grade was negatively correlated with mast cells. Thoracic ultrasonography and airway endoscopy can provide useful information about the inflammatory status of upper and lower airways in the horse.

Feed Intake Parameters of Horses Fed Soaked or Steamed Hay and Hygienic Quality of Hay Stored following Treatment

Horses suffering from equine asthma must consume low-dust forage, with soaking and steaming being suitable methods of hay treatment. The impacts of this treated hay’s subsequent storage and effects on the horses’ chewing activity are largely unknown. Meadow hay was soaked (10–15 °C, 15 min) or steamed (100 °C, 60 min). Microbial counts (colony forming units (CFU)) were determined by culture before and after soaking or steaming, and subsequent storage at 10 and 25 °C for 6, 12 and 24 h (three replicates each). Six horses were fed native, soaked and steamed hay, according to a cross-over design, and chewing parameters were measured. Steaming reduced (p < 0.05) typical mold vs. soaking (0 vs. 50 CFU/g) and yeasts vs. native and steamed hay (0 vs. 102 and 90 CFU/g). Storing soaked hay elevated bacteria, mold, and yeasts (p < 0.05). Within the first 60 min of hay intake, the steamed hay and soaked hay were eaten slower (19.5 and 21.5 g dry matter/min, respectively; p < 0.05) and the steamed hay was chewed more intensely (steamed hay: 3537; native: 2622; and soaked: 2521 chewing cycles/kg dry matter, p < 0.05). Steaming particularly improves the hygienic quality of hay. Soaked hay is not stable when stored and is less accepted by horses.

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 leukocytes by immunocytochemistry and flow cytometry. A multi-color flow cytometric assay was developed to identify leukocytes in equine bronchoalveolar lavage fluid (BALF). Four control and 4 SEA-susceptible horses had BALF collected before and after a 48-hour moldy hay challenge. Antibodies against CD90 uniquely labeled equine neutrophils, and antibodies against CD163 and CD206 identified equine macrophages. Postchallenge AM surface expression of CD163 increased in both groups of horses, but the increase was statistically significant in only the SEA-susceptible group ( P = .02). The surface expression of CD206 on AMs increased significantly in the SEA-susceptible group ( P = .03) but was unchanged in the control group ( P = .5). Increased expression of CD163 and CD206 during exacerbation of SEA suggested an association between AM phenotype and lung inflammation. However, functions of AMs in the pathogenesis of SEA remain to be elucidated.

Some Genetic and Environmental Effects on Equine Asthma in Polish Konik Horses

Current knowledge of the genetic and environmental backgrounds of equine asthma seems to be insufficient, especially for primitive horse breeds. The main objectives of this study were to estimate the effects of sex, birth period, stud, parentage line and inbreeding on asthma morbidity in Polish Konik horses. Records of 274 horses (housed in two studs) were analysed. These animals were allocated to maternal and paternal lines. Individual inbreeding coefficients were extracted from the additive relationship matrix. Horses underwent diagnosis based on observation of the basic symptoms (high frequency of coughing and excessive nasal discharge). Subsequently, some horses (28 individuals) were clinically examined to confirm the earlier observations. Generally, no significant effects of parentage line on heaves morbidity were identified by the use of logistic regression, although the Pearson’s chi-squared test had shown that individuals of some maternal and paternal lines had a predisposition to severe equine asthma. It was concluded that the individual inbreeding level is not associated with the incidence of EA, but a significant effect of the maternal inbreeding coefficient may be observed. It was also found that there is some variability in the incidence of this disease between studs.

Intradermal Testing Results in Horses Affected by Mild-Moderate and Severe Equine Asthma

Equine asthma is an inflammatory respiratory disorder, classified as mild-moderate (MEA) and severe (SEA). SEA is characterized by recurrent exacerbations, consisting of dyspnea, coughing and exercise intolerance; MEA causes poor performance, occasional cough and mucus hypersecretion. Although a precise pathogenesis is not completely understood, allergic mechanisms are considered an important pathophysiological feature of equine asthma. In equine medicine, intradermal testing (IDT) is effective in identifying hypersensitivity to specific allergens. However, to date, the studies about IDT in asthmatic horses obtained contradictory results. This study aims to evaluate IDT responses in MEA and SEA horses and to identify the most significant allergens. Thirty-eight asthmatic horses were enrolled and underwent IDT using 50 allergens; reactions were evaluated at 30 min, 4, 24 and 48 h and were assigned a score from 0 to 4. In SEA horses, the most frequent and strongest reactions were observed at 30 min and 4 h, suggesting the involvement of type I hypersensitivity; in MEA horses, also type IV hypersensitivity seemed to play a major role. Insects, Dermatophagoides spp. and dog epithelium induced in MEA and SEA horses the most significant hypersensitivity responses and could therefore be considered as the main allergenic antigens in our geographic area.