Development of equine upper airway fluid mechanics model for Thoroughbred racehorses

REASON FOR PERFORMING STUDY: Computational fluid dynamics (CFD) models provide the means to evaluate airflow in the upper airways without requiring in vivo experiments. HYPOTHESIS: The physiological conditions of a Thoroughbred racehorse's upper airway during exercise could be simulated. Methods: Computed tomography scanned images of a 3-year-old intact male Thoroughbred racehorse cadaver were used to simulate in vivo geometry. Airway pressure traces from a live Thoroughbred horse, during exercise was used to set the boundary condition. Fluid-flow equations were solved for turbulent flow in the airway during inspiratory and expiratory phases. The wall pressure turbulent kinetic energy and velocity distributions were studied at different cross-sections along the airway. This provided insight into the general flow pattern and helped identify regions susceptible to dynamic collapse. RESULTS: The airflow velocity and static tracheal pressure were comparable to data of horses exercising on a high-speed treadmill reported in recent literature. The cross-sectional area of the fully dilated rima glottidis was 7% greater than the trachea. During inspiration, the area of highest turbulence (i.e. kinetic energy) was in the larynx, the rostral aspect of the nasopharynx was subjected to the most negative wall pressure and the highest airflow velocity is more caudal on the ventral aspect of the nasopharynx (i.e. the soft palate). During exhalation, the area of highest turbulence was in the rostral and mid-nasopharynx, the maximum positive pressure was observed at the caudal aspect of the soft palate and the highest airflow velocity at the front of the nasopharynx. CONCLUSIONS AND CLINICAL RELEVANCE: In the equine upper airway collapsible area, the floor of the rostral aspect of the nasopharynx is subjected to the most significant collapsing pressure with high average turbulent kinetic during inhalation, which may lead to palatal instability and explain the high prevalence of dorsal displacement of the soft palate (DDSP) in racehorses. Maximal abduction of the arytenoid cartilage may not be needed for optimal performance, since the trachea cross-sectional area is 7% smaller than the rima glottidis.     

Effect of a tongue-tie on upper airway mechanics in horses during exercise

OBJECTIVE: To determine the effect of a tongue-tie on upper airway mechanics in exercising horses. ANIMALS: 5 Standardbreds. PROCEDURE: Peak inspiratory and expiratory tracheal and pharyngeal pressures and airflow were measured while horses exercised on a treadmill with and without a tongue-tie. Respiratory rate was also measured. Horses ran at speeds that corresponded to 50 (HR50), 75, 90 (HR90), and 100% of maximal heart rate. The tongue-tie was applied by pulling the tongue forward out of the mouth as far as possible and tying it at the level of the base of the frenulum to the mandible with an elastic gauze bandage. Peak inspiratory and expiratory tracheal, pharyngeal, and translaryngeal resistance, minute ventilation, and tidal volume were calculated. Data were analyzed by use of 2-way repeated-measures ANOVA. For post hoc comparison of significant data, the Student-Newman-Keuls test was used. RESULTS: We were unable to detect significant differences between groups for peak inspiratory or expiratory tracheal or pharyngeal resistance, peak pressure, peak expiratory flow, tidal volume, respiratory rate, or minute ventilation. Horses that ran with a tongue-tie had significantly higher peak inspiratory flows, compared with horses that ran without a tongue-tie. In the post hoc comparison, this effect was significant at 4 m/s, HR50, and HR90. CONCLUSION AND CLINICAL RELEVANCE: Application of a tongue-tie did not alter upper respiratory mechanics in exercising horses and may be beneficial in exercising horses with certain types of obstructive dysfunction of the upper airways. However, application of a tongue-tie does not improve upper airway mechanics in clinically normal horses.     

Factors for prognostic use in equine obstructive small intestinal disease

Twenty horses with small intestinal obstructions requiring surgery were evaluated prospectively. Ten horses lived (group 1) and 10 died (group 2). Eight of the horses in group 1 had simple obstruction and 7 of the horses in group 2 had strangulation obstruction. There was a significant difference (P less than 0.001) between the mean intraluminal hydrostatic pressure in horses of groups 1 and 2 (6.3 cm H2O and 15 cm H2O, respectively). The mean peritoneal fluid protein concentration in horses of groups 1 and 2 (2.8 mg/dl and 5.4 mg/dl, respectively) also differed significantly between groups (P less than 0.01). Histologic evaluation of the intestinal specimens from horses of group 1 (n = 3) and group 2 (n = 6) revealed more severe mucosal lesions in group 2. The measured values that were not significantly different between the 2 groups included PCV, total serum protein content, WBC count, anion gap, and duration of colic before admission. It was concluded that peritoneal fluid protein concentration and intraluminal hydrostatic pressure in small intestinal obstruction may be used as adjuncts to diagnosis and as prognostic indicators.     

3-methylindole as a model of equine obstructive lung disease

3-methylindole was administered orally and intravenously to horses and ponies in order to determine the ability of this chemical to provide a model of equine pulmonary disease. Both routes produced a severe and sometimes fatal pulmonary disease, characterised by bronchiolitis. Clinical signs developed 48 to 72 h after dosing and were most severe between Days 4 and 10 post dosing. Intravenous administration of 3-methylindole produced lung injury more rapidly and at a lower dose rate than the oral route. It is suggested that the respiratory condition induced by this chemical could become a method for standardisation of lung function techniques and interpretation in equine obstructive pulmonary disease.     

Platelets in equine recurrent airway obstruction

Platelets contribute to the pathogenesis of human allergic airway disease. The aim of this study was to compare platelet activating factor (PAF)-induced platelet aggregation and thromboxane (Tx) production, plasma Tx and 5-hydroxytryptamine (5-HT) in ponies with recurrent airway obstruction (RAO), an hypersensitivity to inhaled antigens, and normal ponies, before and after antigen exposure. Plasma 5-HT was significantly higher in ponies with RAO but was not further increased by antigen challenge. There was no difference between PAF-induced platelet aggregation or Tx production, or in plasma Tx before or after challenge. These data suggest there may be a difference between platelet 5-HT uptake in RAO and normal ponies but do not provide evidence of platelet activation following antigen exposure.     

Effect of a tongue-tie on upper airway mechanics during exercise following sternothyrohyoid myectomy in clinically normal horses

OBJECTIVE: To determine the effect of a tongue-tie on upper airway mechanics in clinically normal horses exercising on a treadmill following sternothyrohyoid myectomy. ANIMALS: 6 Standardbreds. PROCEDURE: Upper airway mechanics were measured with horses exercising on a treadmill at 5, 8, and 10 m/s 4 weeks after a sternothyrohyoid myectomy was performed. Pharyngeal and tracheal inspiratory and expiratory pressures were measured by use of transnasal pharyngeal and tracheal catheters connected to differential pressure transducers. Horses were fitted with a facemask and airflow was measured by use of a pneumotachograph. Horses underwent a standardized exercise protocol on a treadmill at 5, 8, and 10 m/s with and without a tongue-tie in a randomized cross-over design. Inspiratory and expiratory airflow, tracheal pressure, and pharyngeal pressure were measured, and inspiratory and expiratory resistances were calculated. RESULTS: We were unable to detect an effect of a tongue-tie on any of the respiratory variables measured. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that a tongue-tie does not alter upper airway mechanics following sternothyrohyoid myectomy in clinically normal horses during exercise.     

Immunological characterization of the equine airway epithelium and of a primary equine airway epithelial cell culture model

Our understanding of innate immunity within the equine respiratory tract is limited despite growing evidence for its key role in both the immediate defense and the shaping of downstream adaptive immune responses to respiratory disease. As the first interface to undergo pathogen invasion, the respiratory epithelium is a key player in these early events and our goal was to examine the innate immune characteristics of equine respiratory epithelia and compare them to an in vitro equine respiratory epithelial cell model cultured at the air-fluid interface (AFI). Respiratory epithelial tissues, isolated epithelial cells, and four-week old cultured differentiated airway epithelial cells collected from five locations of the equine respiratory tract were examined for the expression of toll-like receptors (TLRs) and host defense peptides (HDPs) using conventional polymerase chain reaction (PCR). Cultured, differentiated, respiratory epithelial cells and freshly isolated respiratory epithelial cells were also examined for the expression of TLR3, TLR9 and major histocompatibility complex (MHC) class I and class II using fluorescence-activated cell sorting (FACS) analysis. In addition, cytokine and chemokine profiles from respiratory epithelial tissues, freshly isolated respiratory epithelial cells, and cultured, differentiated, epithelial cells from the upper respiratory tract were examined using real-time PCR. We found that respiratory epithelial tissues and isolated epithelial cells expressed TLRs 1-4 and 6-10 as well as HDPs, MxA, 2'5' OAS, β-defensin-1, and lactoferrin. In contrast, epithelial cells cultured at the AFI expressed TLRs 1-4 and 6 and 7 as well as MxA, 2'5' OAS, β-defensin-1, but had lost expression of TLRs 8-10 and lactoferrin. In addition, MHC-I and MHC-II surface expression decreased in epithelial cells cultured at the AFI compared to isolated epithelial cells whereas TLR3 and TLR9 were expressed at similar levels. Lastly, we found that equine respiratory epithelial cells express an array of pro-inflammatory, antiviral and regulatory cytokines and that after four weeks of in vitro growth conditions, equine respiratory epithelial cells cultured at the AFI retained expression of GM-CSF, IL-10, IL-8, TGF-β, TNF-α, and IL-6. In summary, we describe the development of an in vitro equine respiratory epithelial cell culture model that is morphologically similar to the equine airway epithelium and retains several key immunological properties. In the future this model will be a used to study equine respiratory viral pathogenesis and cell-to-cell interactions.     

Respiratory mechanics in Standardbred horses with sub-clinical inflammatory airway disease and poor athletic performance

The aim of this study was to investigate and quantify respiratory mechanical dysfunctions in Standardbred horses with both poor performance and bronchoalveolar lavage fluid cytology characteristic of inflammatory airway disease (IAD). A control group of healthy Standardbred horses was compared. Respiratory mechanics and breathing pattern were examined at rest and during hyperventilation induced using a rebreathing method. At rest, respiratory mechanics and breathing pattern were superimposable in both groups. In IAD horses, rebreathing increased ventilation, with larger tidal volumes and lower respiratory frequencies. During hyperventilation, IAD animals showed frequency-dependent dynamic lung compliance, and had greater viscous lung resistance and rate of dynamic work of breathing. As IAD alters pulmonary mechanics, the ventilatory load increases and horses requiring significantly higher energy for breathing may suffer restrictions in their athletic performance. This rebreathing method permits early evaluation of respiratory mechanical dysfunction in poorly performing horses with sub-clinical IAD.     

Development and survival of free-living stages of equine strongyles under laboratory conditions

In a series of laboratory studies the optimum conditions for the development and survival of the free-living stages of strongyle parasites occurring in horses in tropical north Queensland were determined. No differences in behaviour were noted between the strongyle species. Development to the infective stage occurred only between 10 and 35 degrees C. The rate was affected by temperature, taking 15-24 days and 3 days, respectively, at the lowest and highest temperatures for the developing stages to reach the infective third stage. Yields of infective larvae were very low outside the range 20-33 degrees C, and were highest at 28 degrees C. Survival of infective larvae was good between 20 and 33 degrees C, and large numbers were recovered after 3 months in faeces incubated at 20-28 degrees C. At 33 and 37 degrees C larval survival was affected by the moisture content of the faeces, with infective larvae surviving better in dry than in moist faeces; even a residual moisture level of 40% significantly reduced the number of larvae recovered from faeces incubated at 37 degrees C for 1 month. Moisture also affected larval development, especially at the higher temperatures of 25-39 degrees C. When faecal moisture content fell to less than or equal to 20% by 3 days, larvae which had not yet reached the infective stage were still pre-infective at 7 days, while all larvae in faeces with adequate moisture had reached the infective third stage. It was not possible to determine the critical faecal moisture level below which larval development ceased, however, 28 degrees C (range 25-33 degrees C) was found to be the optimum temperature. Larval development was very rapid and yields of infective larvae highest at this temperature.     

Influence of subclinical inflammatory airway disease on equine respiratory function evaluated by impulse oscillometry

Reasons for performing study: Inflammatory airway disease (IAD) is a nonseptic condition of the lower respiratory tract. Its negative impact on respiratory function has previously been described using either forced expiration or forced oscillations techniques. However, sedation or drug‐induced bronchoconstriction were usually required. The impulse oscillometry system (IOS) is a noninvasive and sensitive respiratory function test validated in horses, which could be useful to evaluate IAD‐affected horses without further procedures. Objectives: To determine the sensitivity of IOS in detecting alterations of the respiratory function in subclinically IAD‐affected horses without inducing bronchoprovocation and to characterise their respiratory impedance according to frequency for each respiratory phase. Methods: Pulmonary function was evaluated at rest by IOS in 34 Standardbred trotters. According to the cytology of bronchoalveolar lavage fluid (BALF), 19 horses were defined as IAD‐affected and 15 horses were used as control (CTL). Total respiratory resistance (Rrs) and reactance (Xrs) from 1–20 Hz as well as their inspiratory and expiratory components were compared between groups. Results: A significant increase of Rrs at the lower frequencies (R_{1–10 Hz}) as well as a significant decrease of Xrs beyond 5 Hz (X_{5–20 Hz}) was observed in IAD compared to CTL horses. IOS‐data was also significantly different between inspiration and expiration in IAD‐affected horses. In the whole population, both BALF eosinophil and mast cell counts were significantly correlated with IOS measurements. Conclusions: Functional respiratory impairment may be measured, even in the absence of clinical signs of disease. In IAD‐affected horses, the different parameters of respiratory function (Rrs or Xrs) may vary depending on the inflammatory cell profiles represented in BALF. Potential relevance: Impulse oscillometry could be used in a routine clinical setting as a noninvasive method for early detection of subclinical respiratory disease and of the results of treatment in horses.     

Relationship of equine housing to large airway inflammation

The pasture, as compared to stall confinement, has been considered a more desirable environment for maintaining the health of the respiratory system in the horse. This conclusion is based on reports which showed that ventilation in most barns was generally poor, and that horses bedded on straw and fed hay were exposed to many forms of respirable debris. In this study, six normal horses were evaluated for evidence of airway mucosal inflammation after being housed on pasture or stabled in a barn for one month. The response of the horses' airways was measured by assigning scores for the degree of tracheal mucosal secretions that were observed by endoscopic visualization. Cytological examination of transtracheal wash secretions was also performed, as well as histologic evaluation of tracheobronchial tissues obtained by a transendoscopic epithelial biopsy technique. Samples were collected at three time points; the initial collection occurred after the horses were housed on pasture for one month. The horses were subsequently moved to a barn for an equal length of time and samples were obtained at the end of this period. The horses were then returned to their original pasture and final sampling was performed after they were housed in this environment for two months. There were no significant changes in any of the parameters evaluated, regardless of the environment in which the horses were maintained. These findings indicate that housing horses in a barn for four weeks does not cause tracheobronchial mucosal inflammation in a manner that could be detected using the methods employed in this study.     

Expression of thymic stromal lymphopoietin in equine recurrent airway obstruction

Thymic stromal lymphopoietin (TSLP) is a cytokine involved in lymphocyte development. In humans and mice, TSLP drives the differentiation of T helper 2 (Th2) cells and the development of allergic inflammation. The equine TSLP gene has been previously identified and characterized, but its role in the pathogenesis of equine allergic diseases is not known. Our objective was to assess the expression of TSLP in bronchoalveolar lavage (BAL) cells and in primary bronchial epithelial cells (BEC) isolated from horses with recurrent airway obstruction (RAO). RNA was isolated from BAL cells sampled from clinical cases of RAO (n=8) and from control horses (n=12). Furthermore, BAL samples were taken from an additional group of 8 RAO-susceptible and 8 control horses when on pasture (remission) and after 30 days of exposure to moldy hay (exacerbation). In order to study epithelial cells as a potential source of TSLP, cultures of primary bronchial epithelial cells (BEC) were established from 6 RAO-affected and 6 healthy horses and stimulated in vitro with hay dust solution (HDS). Expression of TSLP mRNA was assessed by quantitative real-time RT-PCR (qPCR). Clinical RAO-cases had higher TSLP expression in BAL than control horses (p<0.05). In an experimental group of horses there was no difference between healthy and susceptible horses in remission, whereas after 30-day experimental exposure to moldy hay, all susceptible horses upregulated TSLP expression in BAL (p=0.008, average 6.36-fold increase), whereas in healthy horses there was no significant increase in TSLP expression. BEC generated both from healthy and RAO-affected horses strongly upregulated TSLP expression after 6 h stimulation with HDS, which identifies epithelial cells as potential sources of TSLP in RAO. Finding of increased TSLP expression by BAL cells of RAO-affected horses is in agreement with the contribution of Th2-driven allergic inflammation in the pathogenesis of RAO.     

Miscellaneous conditions of the equine foot

The etiology, clinical presentation, radiographic findings, diagnostic criteria, differential diagnoses, treatment, and prognosis are reviewed for several clinically important conditions of the equine foot. These include pedal osteitis, sheared heels, distal sesamoid bone (navicular) fractures, subchondral bone cyst of the distal phalanx, distal interphalangeal joint subluxation, congenital phalangeal hypoplasia, bipartite and tripartite distal sesamoid bones, keratoma, ossification of the cartilages of the distal phalanx (sidebones), necrosis of the cartilages of the distal phalanx (quittor), thrush, canker, vesicular stomatitis, and chronic selenium toxicosis.