Tissue Doppler Imaging and Two-dimensional Speckle Tracking of Left Ventricular Function in Horses Affected with Recurrent Airway Obstruction before and after Clenbuterol Treatment

Cardiac effects of the β₂-adrenergic agent clenbuterol have been the focus of many studies, but effects on myocardial velocities and myocardial deformation parameters have not yet been evaluated in horses affected with recurrent airway obstruction (RAO) using tissue Doppler imaging (TDI) and two-dimensional speckle tracking (2DST). In our study, 7 horses affected by RAO were treated over 14 days with clenbuterol, 0.8 μg/kg every 12 hours. Standard echocardiographic, TDI (pulsed wave and color TDI), and 2DST examinations were performed before and after the treatment period. Myocardial function was recorded in the right parasternal short-axis view. Percent of fractional shortening and two-dimensional echocardiography (2DE) measurements did not show any significant changes after 2 weeks of treatment. Early diastolic velocity, E, increased significantly after clenbuterol in the left ventricular free wall (LVFW; P = .001). The E/late diastolic velocity (A) quotient (P = .003) and the isovolumetric contractility (P = .035) also increased significantly after treatment. Time parameters, particularly the time interval between the Q-wave in the echocardiograph and atrial release, the time of diastole and Tei index (parameter of global ventricular function), decreased significantly after clenbuterol administration in the LVFW (P = .014/P = .028/P = .015, respectively). The 2DE speckle tracking revealed a significant increase of the early diastolic systolic strain rate (P = .01) in the LVFW after therapy. In conclusion, 2 weeks of treatment with clenbuterol at a dosage of 0.8 μg/kg every 12 hours led to improved cardiac function in severely RAO-affected horses. This could be a sign of myocardial restoration (re-remodeling) after therapy.     

Neutrophil and platelet activation in equine recurrent airway obstruction is associated with increased neutrophil CD13 expression, but not platelet CD41/61 and CD62P or neutrophil–platelet aggregate formation

Recurrent airway obstruction (RAO) in mature horses is characterized by reversible airway obstruction and neutrophilic inflammation; there is also functional activation of circulating platelets and neutrophils. This study was undertaken to determine if changes in activation marker expression and heterotypic aggregate formation can be used as an indicator of this increased functional responsiveness.In vitro conditions for flow cytometric measurement of CD13, CD41/61 and CD62P expression on activated cells and heterotypic aggregate formation were established. Values were then compared before and after antigen challenge of RAO and healthy horses. Platelet adhesion to serum-coated plastic was measured as a functional marker of platelet activation.In vitro activation resulted in increased expression of neutrophil CD13 and platelet CD41/61 and CD62P. Activation of both cell types caused a significant increase in neutrophil–platelet aggregates.In horses with RAO, but not controls, there was a significant increase in the percentage of CD13 positive neutrophils at 10 h and 24 h and in the mean fluorescence intensity at 10 h. This was accompanied at 24 h by an increased mean platelet side scatter and thrombin-stimulated platelet adhesion.In conclusion, CD13 expression can be used as an indicator of equine neutrophil activation both in vitro and in vivo. Equine platelet activation in vitro can be detected by measuring CD41/61 or CD62P expression, and PAF-activated platelets and neutrophils form aggregates. However, despite evidence of circulating platelet activation, neither a change in expression of platelet activation markers, nor heterotypic aggregate aggregate formation could be detected.     

The Effect of Aerosolized and Intravenously Administered Clenbuterol and Aerosolized Fluticasone Propionate on Horses Challenged with Aspergillus fumigatus Antigen

β-Agonists have been shown to display anti-inflammatory properties in several experimental models. The aim of this study was to investigate the anti-inflammatory properties of clenbuterol (CB), administered either intravenously or by aerosol, in comparison with fluticasone propionate (FP) in recurrent airway obstruction (RAO)-susceptible horses. Eight horses, of which five were known to be susceptible to RAO, underwent an inhalation challenge with Aspergillus fumigatus (AF) antigen and were treated with CB intravenously, CB by aerosol, or FP by aerosol. Twenty-four hours after the challenge, bronchoalveolar lavage was performed, the total and differential cell counts were assessed, and cytokines were measured in isolated alveolar macrophages. After challenge with AF, RAO-susceptible horses showed an increase in total cell count, based on an increase in macrophages and lymphocytes, which was inhibited by treatment with intravenous CB, aerosolized CB and aerosolized FP. Neutrophil ratios were decreased when treated with aerosolized CB and FP. Expression of interleukin (IL)-1β and IL −8 was significantly increased after AF challenge .Interleukin −1β was significantly decreased following treatment with intravenous CB, aerosolized CB and aerosolized FP, whereas only FP decreased the expression of IL-8. These data suggest that the anti-inflammatory property of CB provide new opportunities in the therapeutic intervention of early inflammation in RAO.     

Equine recurrent airway obstruction and insect bite hypersensitivity: understanding the diseases and uncovering possible new therapeutic approaches

Recurrent airway obstruction (RAO) and insect bite hypersensitivity (IBH) are allergic conditions that are commonly encountered in the horse. Whilst complete allergen avoidance is an effective management strategy for both diseases, this may not be achievable in all cases and treatment options are therefore required. The inflammatory response is the main therapeutic target for glucocorticoids given to horses with RAO and severe cases of IBH, whilst the bronchodilators used in RAO primarily target airway smooth muscle. Such drugs are effective in most but not all individuals and there may be unwanted adverse effects. This article will review how knowledge of drug action and the pathogenesis of RAO and IBH can be utilised to identify potential targets for novel therapeutic agents that, in the longer term, may be safer and/or more effective in managing the allergic horse.     

Influence of valvular insufficiency and recurrent airway obstruction on haemodynamics and therapy in warmblood horses with atrial fibrillation

The aim of this study was to investigate the potential haemodynamic effects of valvular insufficiency and recurrent airway obstruction (RAO) in horses with atrial fibrillation (AF). Therefore in ten healthy horses (group 1) and 40 horses with AF a clinical examination, a lung examination, echocardiography and right heart catheterization for measurement of intracardic and pulmonary pressures were performed. According to the clinical findings the horses with AF were subdivided into 4 groups (group 2: AF; group 3: AF/valvular insufficiency; group 4: AF/RAO; group 5: AF/valvular insufficiency/RAO). Most of the horses of group 3 and 5 suffered from two valvular insufficiencies (mitral and tricuspid valve insufficiency: n=11, mitral and aortic valve insufficiency: n=2). The remaining horses showed a single mitral (n=6), tricuspid (n=2) or aortic valve insufficiency (n=1) or more than two valvular insufficiencies (n=4). In group 2 right ventricular mean pressure (RVPm) was higher than in group 1 and 4 (P<0.025); diastolic right ventricular pressure was higher than in group 1; PWP was higher than in group 1 and group 4; PDP was lower compared to group1. Compared to group1 in group 3 left atrial diameter (LA) was greater; the PAPs was higher and the PDP lower (P<0.05). In group 4 RVPm and PWP was lower compared to group 2. In group 5 LA, fractional shortening and diastolic left ventricular diameter were greater, PWP and PAPs were higher and PDP lower compared to group1. Twenty six of the 40 horses with AF (65%) were treated. Successful cardioversion to sinus rhythm occurred in 15 horses (58%). Therapy was successful in 50% of the treated horses of group 2 and 3, in 67% of the treated horses of group 4 and in 63% of the treated horses in group 5. In conclusion the presence of valvular insufficiency or RAO influences the haemodynamics of horses with AF.     

Effects of a MAPK p38 inhibitor on lung function and airway inflammation in equine recurrent airway obstruction

Reasons for performing study: It has been suggested that many of the beneficial effects of corticosteroids are mediated through mitogen‐activated protein kinase (MAPK) p38 inhibition. Objective: To investigate the efficacy of the MAPK p38 inhibitor compound MRL‐EQ1 to either prevent (Phase 1) or treat (Phase 2) recurrent airway obstruction (RAO) in horses. Methods: MRL‐EQ1 was administered i.v. at a dosage of 0.75‐1.5 mg/kg bwt q. 12 h. In Phase 1, susceptible horses in clinical remission were divided into 2 groups (n = 5/group), based on historical values of respiratory mechanics. All horses were entered in the study in pairs (one control, one treated horse) and exposed to the same environmental challenge (stabling, mouldy hay and dusty conditions). The treatment group received MRL‐EQ1 for 14 days while the control horses were untreated during the same period. In Phase 2, affected horses were ranked by severity of respiratory dysfunction and split randomly into either dexamethasone or MRL‐EQ1 treatment groups (n = 5/group). Bronchoalveolar lavage fluid, respiratory mechanic measurements, MRL‐EQ1 plasma concentration and tumour necrosis factor (TNF) whole blood activity were evaluated sequentially. Results: In Phase 1, MRL‐EQ1 did not prevent the occurrence of clinical signs and pulmonary inflammation. However, treatment was associated with a reduction in severity and a delay in the onset of signs and a reduction in pulmonary neutrophilia. In Phase 2, plasma concentrations achieved resulted in ex vivo suppression of lipopolysaccharide‐induced TNF production in equine blood. MRL‐EQ1 did not improve airway inflammation or lung function and was associated in a dose dependent manner with behavioural (depression, excitability) and blood changes (neutrophilia, increased serum muscle enzyme concentrations). Conclusions: Inhibition of p38 in the horse was partially effective in reducing clinical signs and airway inflammation when administered prior to, but not during clinical exacerbation in RAO. Potential relevance: Inhibitors of p38 MAPK with a better toxicity profile may be effective in the prevention or treatment of RAO.     

Oxidant‐Antioxidant Status in the Blood of Horses with Symptomatic Recurrent Airway Obstruction (RAO)

Background

Systemic oxidative stress in horses with recurrent airway obstruction (RAO) is poorly characterized.

Objectives

The goal of this study was to investigate whether equine RAO is associated with systemic disturbances in the oxidant‐antioxidant equilibrium.

Animals

Seven healthy horses and 7 horses with symptomatic RAO.

Methods

A prospective study. Healthy and RAO‐affected horses were exposed to a 48‐hour challenge with moldy hay and straw to induce clinical exacerbation of RAO. Venous blood was collected and the activities of the superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione reductase (GR) in equine erythrocyte lysates were measured. The concentration of thiobarbituric acid‐reactive substances (TBARSs) was assessed both in erythrocyte lysates and in plasma.

Results

A significant increase in the activities of GPx and SOD was detected in RAO‐affected horses compared with the control animals. There was no significant difference between groups in terms of the erythrocyte lysate activities of CAT, GR, or TBARs or the plasma concentration of TBARs.

Conclusion and Clinical Importance

Our results support the hypothesis that RAO in horses is associated with systemic oxidative stress. Future studies are needed to assess whether horses suffering from RAO can benefit from antioxidant supplementation.     

Analysis of Canadian and Irish forage,oats and commercially available equine concentrate feed for pathogenic fungi and mycotoxins

Respiratory infections, recurrent airway obstruction (RAO) and exercise induced pulmonary haemorrhage (EIPH) are major causes of poor performance in horses. Fungi and mycotoxins are now recognised as a major cause of these conditions. The most notable fungi are Aspergillus and Fusarium. Fungal spores can originate from forage, bedding and feed and, in turn, these fungal spores can produce a series of mycotoxins as secondary metabolites.This study set out to ascertain the degree of fungal and mycotoxin contamination in feed and fodder used in Irish racing yards over a one-year period. Weather conditions in forage producing areas were sampled by Met Eireann and the Canadian Meteorological Service.Fifty per cent of Irish hay, 37% of haylage and 13% of Canadian hay contained pathogenic fungi. Of the mycotoxins, T2 and zearalenone were most prominent. Twenty-one per cent of Irish hay and 16% of pelleted feed contained zearalenone. Forty per cent of oats and 54% of pelleted feed contained T2 toxins.