Comparative efficacy of inhaled albuterol between two hand-held delivery devices in horses with recurrent airway obstruction

Reasons for performing study: Studies investigating the clinical efficacy of albuterol administered with the same propellant and commercially available delivery devices in horses with recurrent airway obstruction (RAO) are not currently available. Objectives: To determine the efficacy of aerosolised albuterol administered to horses with RAO by means of 2 commercially available, hand‐held delivery devices. Methods: Ten horses with RAO were kept in a dusty environment and fed mouldy hay to induce airway obstruction. Lung mechanics were measured before and after the procedure. ΔP_max was measured 5 min after administration of 180 µg of albuterol from a pressurised metered dose inhaler, using an aerosol delivery device chosen randomly. This process was repeated every 5 min until maximal bronchodilation was achieved. After a 24 h washout period, lung mechanics data were again collected using the other aerosol delivery device. Results: Aerosolised albuterol induced a significant and rapid bronchodilation in the horses using both aerosol delivery devices. No statistically significant difference in pulmonary function was observed in response to albuterol therapy between the 2 devices. The dose required to achieve 50% of maximal bronchodilation was not statistically different between the 2 devices (173.35 ± 78.35 µg with Device 1 and 228.49 ± 144.99 µg with Device 2, P = 0.26). The decrease in lung resistance tended to be more pronounced after albuterol administration with Device 1 (P = 0.066). Conclusions: Aerosolised albuterol is an effective bronchodilator in horses with recurrent airway obstruction. There is no statistically significant difference between the 2 commercially available aerosol delivery devices in terms of efficacy. Potential relevance: Aerosolised albuterol is effectively delivered using currently available devices leading to maximal bronchodilation in horses with RAO at an average dose of 540 µg.     

Lung function and airway cytologic profiles in horses with recurrent airway obstruction maintained in low-dust environments

BACKGROUND: The effects of long-term environmental management on airway obstruction and inflammation in horses with recurrent airway obstruction (RAO) are unknown. HYPOTHESIS: Horses with RAO maintained in low-dust environments have persistent airway obstruction and neutrophilic inflammation. ANIMALS: Study horses were treated for RAO and then maintained in low-dust environments with no medical management. Horses were classified into 3 groups by years after diagnosis: 1 year (time 1, n = 9), 2-3 years (time 2, n = 7), and 5-6 years (time 3, n = 8). The comparison groups were age-matched healthy horses. METHODS: In this cross-sectional study, a clinical examination was performed, and the clinical score was calculated. Standard lung function, forced expiratory maneuvers, and the cytology of bronchoalveolar lavage fluid (BALF) were evaluated. RESULTS: The clinical scores of the RAO horses were higher than those of the non-RAO horses at time 2 (P = .018). Standard lung function data were not different between the groups at any time point. The forced expiratory flow between 75-95% of exhaled vital capacity was lower in RAO horses than in non-RAO horses at all time points (P < .02), indicating persistent peripheral airway obstruction. Cytologic evaluation of BALF revealed no difference in total nucleated cell numbers or differential cell counts between RAO and non-RAO horses at any time point. CONCLUSIONS AND CLINICAL IMPORTANCE: The peripheral airway obstruction detected in horses with RAO maintained in low-dust environments likely is due to irreversible airway remodeling but is not associated with cytologic evidence of airway inflammation.     

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.     

Differential association of MUC5AC and CLCA1 expression in small cartilaginous airways of RAO‐affected and control horses

Reasons for performing study: Airway mucus accumulation is associated with indoor irritant and allergen exposure in horses with recurrent airway obstruction (RAO). Epidermal growth factor receptor (EGFR) and a chloride channel (calcium activated, family member 1; CLCA1) are key signalling molecules involved in mucin gene expression. Objectives: We hypothesised that exposure to irritants and aeroallergens would lead to increased expression of the mucin gene eqMUC5AC and increased stored mucosubstance in the airways of RAO‐affected horses, associated with increased neutrophils and CLCA1 and EGFR mRNA levels. Methods: We performed quantitative RT‐PCR of eqMUC5AC, CLCA1 and EGFR; volume density measurements of intraepithelial mucosubstances; and cytological differentiation of intraluminal inflammatory cells in small cartilaginous airways from cranial left and right and caudal left and right lung lobes of 5 clinically healthy and 5 RAO‐affected horses that had been exposed to indoor stable environment for 5 days before euthanasia. Results: Neutrophils were increased in RAO‐affected horses compared to clinically healthy controls. EqMUC5AC mRNA levels were positively correlated with both CLCA1 and EGFR mRNA levels in RAO‐affected horses but only with CLCA1 in controls. The relationship between eqMUC5AC and CLCA1 differed in the 2 groups of horses with RAO‐affected animals overexpressing CLCA1 in relation to eqMUC5AC. Conclusions: These data implicate CLCA1 as a signalling molecule in the expression of eqMUC5AC in horses but also suggest differential regulation by CLCA1 and EGFR between horses with RAO and those with milder degrees of airway inflammation.     

Efficacy of oral prednisolone and dexamethasone in horses with recurrent airway obstruction in the presence of continuous antigen exposure

Reasons for performing study: Orally administered prednisolone and dexamethasone are used commonly in the treatment of recurrent airway obstruction (RAO) in horses. However, the efficacy of prednisolone in improving pulmonary function during continuous antigen exposure has not been evaluated critically and there is little evidence supporting the efficacy of low‐dose oral dexamethasone in the same conditions. Hypothesis: Oral prednisolone and dexamethasone improve pulmonary function in RAO under conditions of continuous antigen exposure, and dexamethasone is more effective than prednisolone at commonly used dosages. Methods: Using a randomised crossover design, prednisolone (2 mg/kg bwt) and dexamethasone (0.05 mg/kg bwt) were administered per os, s.i.d. for 7 days, to 7 horses during clinical exacerbation of the disease. Maximal difference in transpulmonary pressure (ΔP_L), lung resistance (R_L) and elastance (E_L) were measured before and after 3 and 7 days of treatment. Results: Prednisolone and dexamethasone improved pulmonary function significantly. However, the improvement was of greater magnitude after 3 and 7 days of treatment with dexamethasone compared to prednisolone. Also, after 7 days of treatment with dexamethasone, ΔP_L and R_L were not different from values obtained when horses were on pasture, while all 3 pulmonary function parameters remained different from pasture values after prednisolone treatment. Conclusions: Both corticosteroids improve pulmonary function, in spite of continuous antigen exposure. However, oral dexamethasone at 0.05 mg/kg bwt is more effective than prednisolone at 2 mg/kg bwt in the treatment of RAO. Potential relevance: Prednisolone was shown, for the first time, to our knowledge, to improve the pulmonary function of horses with RAO in the presence of continuous antigen exposure. This study also demonstrates the efficacy of low‐dose oral dexamethasone in reversing airway obstruction in these conditions.     

Abnormalities in lung surfactant in horses clinically affected with recurrent airway obstruction (RAO)

Background: Abnormalities in lung surfactant are well described in human respiratory diseases including asthma, but are poorly described in horses. Hypothesis: Lung surfactant is abnormal in horses with clinical signs of recurrent airway obstruction (RAO). Animals: Six healthy horses and 5 horses with RAO. Methods: Bronchoalveolar lavage fluid (BALF) was obtained from all horses by standard procedures. Cell‐free BALF was separated into crude surfactant pellets (CSP) and supernatant via ultracentrifugation. Phospholipid and protein content was analyzed from both of these fractions. Phospholipid composition of CSP was determined using high‐performance liquid chromatography with an evaporative light scatter detector. Surface tension of CSP was measured with a pulsating bubble surfactometer. Results: Compared with healthy horses, surfactant from RAO‐affected horses was characterized by significantly decreased phospholipid content in total surfactant (median; range: 23.2; 14.7–62.2 μg/mL BALF versus 172; 111–267 μg/mL BALF, P= .0062) and CSP (20.2; 6.4–48.9 μL/mL BALF versus 155; 94.4–248 μg/mL BALF, P= .0062), and a significantly lower percentage of phosphatidylglycerol (PG) (4.5; 3.6–5.6% versus 6.6; 4.1–7.6%, P= .028). Furthermore, the ratio between the percentages of phosphatidylcholine and PG was significantly higher in RAO‐affected horses than in healthy horses (20.9; 16.6: 25.9 versus 13.9; 11.8–22.8, P= .045). Conclusions and Clinical Importance: This study demonstrates that surfactant from RAO‐affected horses is abnormal. Further studies are needed to determine if these abnormalities are related to an increased tendency for bronchoconstriction and to a decreased ability to clear airway mucus in RAO‐affected horses.     

Evaluation of leukotriene biosynthetic capacity in lung tissues from horses with recurrent airway obstruction

OBJECTIVE: To evaluate leukotriene (LT) biosynthetic capacity in lung tissue from healthy horses and horses with recurrent airway obstruction (RAO). SAMPLE POPULATION: Lung parenchyma and airway specimens from 8 RAO-affected and 5 healthy horses. PROCEDURE: Horses were stabled for > or = 72 hours. Blood was drawn before euthanasia, after which lung specimens were collected. Tissue strips from small airways and parenchyma were incubated in organ baths with the precursor LTA4 or stimulated with calcium ionophore A23187 or the tripeptide N-formyl-Met-Leu-Phe (fMLP), with or without exogenous arachidonic acid, in the presence of isolated blood neutrophils. RESULTS: Stabling induced typical clinical signs of airway obstruction in RAO-affected horses but not control horses. When lung parenchyma or airway specimens from both groups of horses were incubated with calcium ionophore, with or without arachidonic acid, they did not form LT. In contrast, addition of LTA4 to both tissues resulted in conversion to LTB4, although concentrations of LTC4 were negligible in airways and parenchymal strips from healthy and RAO-affected horses. Incubation of airway and parenchymal strips with suspensions of autologous neutrophils did not influence formation of LT stimulated by calcium ionophore or fMLP, with or without exogenous arachidonic acid. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that lung parenchyma and airway tissues themselves are not of substantial importance for LT formation in the lungs, although these tissues possessed some LTA4 hydrolase activity, enabling LTB4 formation. It may be speculated that LTB4 originates primarily from neutrophils and may play a role in the inflammatory events of RAO.     

Evaluation of a risk-screening questionnaire to detect equine lung inflammation: results of a large field study

Reasons for performing study: The diagnosis of equine recurrent airway obstruction (RAO) and inflammatory airway disease (IAD) is based on clinical signs and increased inflammatory cell percentages in the bronchoalveolar lavage (BAL) fluid. Since a BAL is an invasive procedure, a risk‐screening questionnaire (RSQ) would be a valuable screening tool for lung inflammation. Objective: To evaluate the accuracy of a RSQ to detect lower airway inflammation (LAI) in a large population of horses. Methods: A standardised BAL was performed in the field on 167 horses in Alberta, Canada. Horses were separated into 3 categories: 1) BAL normal; 2) BAL mild to moderate LAI (MLAI), and 3) BAL severe LAI (SLAI). The horse owners were asked to complete a RSQ. The RSQ scores were compared to the BAL results to determine the likelihood of a horse having MLAI, SLAI or no LAI. Results: Based on BAL cytology, 28 (17%) horses were normal and 139 (83%) were abnormal, with 110 (66%) showing MLAI and 29 (17%) SLAI. Horses with SLAI and MLAI had a mean RSQ score of 0.95 and 0.70, respectively, compared to 0.60 for normal BAL horses. Horses with SLAI showed more clinical signs than normal and MLAI horses. The sensitivity and negative predictive values of the RSQ for detecting SLAI using a cut‐off score of 0.87, were excellent at 0.90 (95%CI 0.73–0.98) and 0.96 (95%CI 0.82–1.00). Questions on the clinical signs typically found in RAO cases differed significantly between horses with BAL SLAI and those with BAL normal. Conclusions: Prevalence of MLAI was high in this population. Although the RSQ did not allow differentiating normal horses from horses with MLAI, it has a high sensitivity to detect horses with SLAI and is therefore a good screening tool for SLAI.     

Randomized, controlled study of inhaled fluticasone propionate, oral administration of prednisone, and environmental management of horses with recurrent airway obstruction

OBJECTIVE: To determine whether administration of glucocorticoids provides additional benefits to environmental management of horses with recurrent airway obstruction (RAO). ANIMALS: 28 horses with RAO. PROCEDURE: Horses were classified as having mild, moderate, or severe RAO. Within each category, horses were randomly assigned to receive inhaled fluticasone propionate, inhaled control substance, or oral administration of prednisone. During the 4-week study, horses were maintained outdoors and fed a pelleted feed. Clinical scores, pulmonary function, results of cytologic examination of bronchoalveolar lavage fluid (BALF), and adrenal gland function were determined before and 2 and 4 weeks after initiation of treatment. RESULTS: Clinical score and pulmonary function of all RAO-affected horses improved during the treatment period. After 4 weeks, clinical scores and pulmonary function of horses treated with a glucocorticoid were not different from those for the control treatment. In horses with severe RAO, treatment with fluticasone for 2 weeks resulted in significantly greater improvement in pulmonary function, compared with pulmonary function after treatment with prednisone or the control substance. Treatment with a glucocorticoid for 4 weeks and a low-dust environment did not have any effect on cellular content of BALF Treatment with prednisone for 2 weeks resulted in a significant decrease in serum cortisol concentration, compared with concentrations after administration of fluticasone or the control substance. CONCLUSIONS AND CLINICAL RELEVANCE: Environmental management is the most important factor in the treatment of horses with RAO. Early treatment with inhaled fluticasone can help accelerate recovery of horses with severe RAO.     

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.     

Neurokinin receptors in recurrent airway obstruction: A comparative study of affected and unaffected horses

The purpose of the study was to compare in vitro airway responses to neurokinin A & B (NKA and NKB) and expression of NK-2 receptors in airways of horses affected and unaffected with recurrent airway obstruction (RAO). Neurokinin-A, an inflammatory mediator belonging to the tachykinin family of neuropeptides, causes bronchoconstriction by binding to NK-2 receptors. Neurokinin-B is a lesser-known neuropeptide that acts on NK-3 receptors. Horses were placed into RAO-affected and RAO-unaffected groups based on their history, clinical scoring, and pulmonary function testing. Lung tissue from each lobe was collected for immunohistochemical staining for NK-2 receptors. Cumulative concentration-response relationships were determined on bronchial rings (4-mm wide) collected and prepared from the right diaphragmatic lung lobe to graded concentrations (half log molar concentrations 10⁻⁷M to 10⁻⁴M) of NKA and NKB. The results showed that NKA caused significantly greater contraction than NKB in both groups. In RAO-affected horses, both agents produced significantly greater bronchial contractions than those in the RAO-unaffected horses. Immunohistochemical staining showed that the overall NK-2 receptor distribution was significantly increased in bronchial epithelium and smooth muscles of bronchi and pulmonary vessels of RAO-affected than RAO-unaffected horses. The findings indicate that NK-2 receptors are up-regulated in RAO, suggesting that NK-2 receptor antagonists may have some therapeutic value in controlling the progression of airway hyperreactivity in horses affected with RAO.     

Comparison of the caudal lung borders determined by percussion and ultrasonography in horses with recurrent airway obstruction

The aim of the study was to evaluate the diagnostic value of thoracic percussion and ultrasonography with the help of distance measurements and statistical methods in the determination of the caudal lung border in horses with recurrent airway obstruction (RAO). Examinations were performed on 11 healthy, warm-blooded horses of different breeds, age and grade of disease. First, the caudal lung border was determined by the traditional indirect percussion method in the 10th, 12th, 14th and 16th intercostal spaces at the end of inspiration and expiration on both sides of the thorax. To apply standardised measurements, a fix point was chosen as described earlier by the same authors for healthy horses. The distance between this point and the caudal lung border was measured with a tape-measure. Percussion was followed by ultrasonographic determination of the caudal lung border. Measurements were performed in the same way as described for the percussion technique. Mean values and standard errors of absolute values of differences between percussion and ultrasonographic measurements were the following, in centimetres (10th, 12th, 14th and 16th intercostal spaces). Left side expiration: 1.4, 0.4; 0.8, 0.2; 0.9, 0.2; 0.8, 0.4; left side inspiration: 0.8, 0.3; 1.5, 0.3; 1.4, 0.3; 1.1, 0.3; right side expiration: 2.1, 1.0; 2.1, 0.5; 1.6, 0.5; 0.8, 0.1; right side inspiration: 1.5, 0.7; 1.2, 0.6; 0.8, 0.2; 0.8, 0.3, respectively. Ultrasonography proved to be reliable in determining the caudal lung borders in horses with RAO. Results of the percussion examination did not differ significantly from those of the ultrasound method which was used as a reference technique. The differences between inspiration and expiration were greater in horses with RAO than in healthy horses in a previous study. Based on these results, percussion can be used as an integrated part of the physical examination in diagnosing caudal shift of the caudal lung border of horses suffering from RAO.     

Measurement of tidal breathing flow-volume loop indices in horses used for different sporting purposes with and without recurrent airway obstruction

Tidal breathing flow-volume loop (TBFVL) indices in healthy horses and in horses suffering from mild and moderate to severe recurrent airway obstruction (RAO) were measured. In addition, the ability of these indices to differentiate healthy controls from horses with mild RAO by separating the horses according to their usage was investigated. The horses were assigned to three sporting categories: 1 Leisure horses; 2 Horses competing at events with a duration of more than five minutes (eventing, coaching and endurance); and 3 Horses competing at events with a duration of less than five minutes (dressage and show jumping). Flow-volume curves were recorded with an ultrasonic spirometer and TBFVL indices were calculated with specially dedicated software. The sporting usage of the horses had a significant effect on the differentiating ability of TBFVL indices in diagnosing different degrees of RAO. The index TIF50 differentiated control horses from horses with minor RAO in all sporting categories.     

Relationship of horse owner assessed respiratory signs index to characteristics of recurrent airway obstruction in two Warmblood families

Reasons for performing study: The horse owner assessed respiratory signs index (HOARSI‐1–4, healthy, mildly, moderately and severely affected, respectively) is based on owner‐reported clinical history and has been used for the investigation of recurrent airway obstruction (RAO) genetics utilising large sample sizes. Reliable phenotype identification is of paramount importance in genetic studies. Owner reports of respiratory signs have shown good repeatability, but the agreement of HOARSI with an in‐depth examination of the lower respiratory tract has not been investigated. Objectives: To determine the correlation of HOARSI grades 3/4 with the characteristics of RAO and of HOARSI‐2 with the characteristics of inflammatory airway disease. Further, to test whether there are phenotypic differences in the manifestation of lung disease between families. Methods: Seventy‐one direct offspring of 2 RAO‐affected Warmblood stallions (33 from the first family, 38 from the second) were graded as HOARSI‐1–4 and underwent a clinical examination of the respiratory system, arterial blood gas analysis, endoscopic mucus scoring, cytology of tracheobronchial secretion (TBS) and bronchoalveolar lavage fluid (BALF), and clinical assessment of airway reactivity to methacholine chloride. Results: HOARSI‐3/4 animals in clinical exacerbation showed signs consistent with RAO: coughing, nasal discharge, abnormal lung sounds and breathing pattern as well as increased numbers of neutrophils in TBS and BALF, excessive mucus accumulation and airway hyper‐responsiveness to methacholine. HOARSI‐3/4 horses in remission only had increased amounts of tracheal mucus and TBS neutrophil percentages. Clinical phenotypes were not significantly different between the 2 families. Conclusions and clinical relevance: HOARSI reliably identifies RAO‐affected horses in our population.     

Aerosolized albuterol sulfate used as a bronchodilator in horses with recurrent airway obstruction.

OBJECTIVE: To determine the dose of aerosolized albuterol sulfate required to cause bronchodilation in horses with recurrent airway obstruction (RAO) and duration of this effect. ANIMALS: 19 horses with RAO (10 in experiment 1; 9 in experiment 2). PROCEDURE: Horses were moved from pasture to stables, and airway obstruction was induced. Pulmonary function was measured in 10 horses before and 5, 10, and 30 minutes after administration of vehicle or 120, 240, 360, or 720 microg of the drug. Nine horses received vehicle or 360 or 720 microg of albuterol, and pulmonary function was measured at baseline and 5 minutes and 1, 2, 3, 4, 5, 6, and 7 hours later. Horses were evaluated for adverse drug effects. RESULTS: 360 microg of albuterol was required to cause significant bronchodilatation; 720 microg did not enhance bronchodilatation or increase duration of action. Depending on which pulmonary function parameter was evaluated, bronchodilatation achieved by use of albuterol lasted between 30 minutes and 1 hour. Because there was a significant vehicle effect, the combined effect of vehicle and drug lasted up to 3 hours. Adverse effects were not observed. CONCLUSIONS: Aerosolized albuterol, 360 or 720 microg, is a safe and effective bronchodilator in horses with RAO. Onset of action is rapid (5 minutes), and effects last from 30 minutes to 3 hours. CLINICAL RELEVANCE: Aerosolized albuterol is useful for treatment of bronchospasm in horses with RAO.     

Expression of toll‐like receptor 2 mRNA in bronchial epithelial cells is not induced in RAO‐affected horses

Reasons for performing study: Airway inflammation in recurrent airway obstruction (RAO) is triggered by housing affected horses in stables. It has been suggested that RAO is an allergic condition, but innate immune mechanisms are also involved. Fungal products activate innate immune mechanisms through toll‐like receptor 2 (TLR2). In human airway epithelium, TLR2 activation leads to interleukin (IL)‐8 production. This pathway is negatively regulated by the zinc finger protein A20. This study was performed to enhance understanding of innate immune mechanisms in RAO. Hypothesis: TLR2 and IL‐8 mRNA are elevated in RAO during stabling compared with controls. A20 mRNA is negatively associated with the numbers of airway inflammatory cells. Objectives: To determine TLR 2 , IL‐8 and A20 mRNA expression in lungs of stabled and pastured RAO‐affected and control horses. Methods: Airway obstruction and inflammatory cell counts in bronchoalveolar lavage were measured, and TLR 2 , IL‐8 and A20 mRNA expression quantified by qRT‐PCR in 6 RAO‐affected and 6 control horses, during and after exposure to hay and straw. Results: Airway obstruction and neutrophils were increased in RAO‐affected horses during stabling. While stabling increased IL‐ 8 , TLR2 and A20 mRNA were unaffected. TLR2 and A20 were significantly correlated (r = 0.83) and A20 mRNA was negatively associated with inflammatory cells. Potential relevance: Stabling does not lead to an increase in TLR2 expression. Other molecules or processes in the TLR2 cascade might be important in fungal‐induced airway inflammation. Equine epithelial‐derived A20 may be involved in modulation of airway inflammation.     

Analysis of risk factors for recurrent airway obstruction in North American horses: 1,444 cases (1990-1999)

OBJECTIVE: To identify risk factors for recurrent airway obstruction (RAO) among horses examined at veterinary teaching hospitals in North America. DESIGN: Retrospective case-control study. ANIMALS: 1,444 horses with RAO and 1,444 control horses examined for other reasons. PROCEDURE: The Veterinary Medical Database was searched for records of horses in which RAO was diagnosed. A control group was identified by randomly selecting a horse with a diagnosis other than RAO that matched the institution and year of admission for each of the horses with RAO. Information obtained included hospital, admission year and month, age, sex, breed, and discharge status. The association between risk factors and diagnosis of RAO was estimated with logistic regression models. RESULTS: The risk of RAO increased significantly with age, with horses > or = 7 years old being 6 to 7 times as likely to have RAO as were horses < or = 4 years old. Thoroughbreds were 3 times as likely to have RAO as were ponies. Horses were 1.6 and 1.5 times as likely to be examined because of RAO during winter and spring, respectively, than they were during summer. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that RAO was more likely to be diagnosed in females, horses > or = 4 years old, and Thoroughbreds and that RAO has a seasonal distribution.     

Environment and prednisone interactions in the treatment of recurrent airway obstruction (heaves)

Recurrent airway obstruction (RAO) or heaves is a manifestation of a hypersensitivity to dust, moulds, and spores in the environment of a susceptible horse. Although in the majority of RAO-affected horses, clinical remission can be achieved by keeping horses at pasture to reduce their allergen exposure, this often is not practicable. For this reason, we investigated if changing the environment of a single stall in a 4 stall stable was sufficient to improve lung function and reduce inflammation in RAO-affected horses. In addition, we determined if addition of oral prednisone provided additional benefit. Twelve RAO-susceptible horses were stabled, fed hay, and bedded on straw until they developed airway obstruction. At this point, bedding was changed to wood shavings and they were fed a pelleted diet for 2 weeks. Lung function was measured and bronchoalveolar lavage was performed before and 3, 7, and 14 days after environmental modification. In a crossover design, horses were treated for the 14 days with prednisone tablets (2.2 mg/kg bwt, q. 24 h). Horses then returned to pasture for 30 days. Airway obstruction was greatest before environmental modification. Significant improvement in lung function occurred within 3 days of the change in environment and continued to Day 7. Airway function was best after 30 days at pasture. The clinical response achieved by environmental modification was not significantly improved by addition of oral prednisone. The total number of cells, total neutrophils, and percent neutrophils was greatest before environmental modification. In the absence of prednisone, total and percent neutrophils did not decrease until Day 14 and total cell number until 30 days at pasture. In the presence of prednisone, total cells and total and percent neutrophils decreased by Day 3 and again at pasture. The fact that lung function can be improved within 3 days by environmental management alone emphasises the need for allergen reduction as the cornerstone of treatment of RAO. Although prednisone induced a more rapid reduction in airway inflammation, this was not associated with a more rapid improvement in airway function.     

Theophylline does not potentiate the effects of a low dose of dexamethasone in horses with recurrent airway obstruction

REASONS FOR PERFORMING STUDY: Theophylline has been shown to have corticosteroid-sparing effects for the treatment of human asthma. A similar effect, if present in horses, would allow diminishing the dose of corticosteroids administered to equine patients with inflammatory airway diseases. OBJECTIVES: To evaluate whether theophylline potentiates the effects of a low dose of dexamethasone when treating horses with recurrent airway obstruction (RAO). HYPOTHESIS: Theophylline has steroid-sparing effects in horses with RAO. METHODS: Ten mature mixed breed horses in clinical exacerbation of RAO were studied. Using an incomplete crossover design and 3 experimental periods of 7 days duration, horses were distributed randomly in 5 treatment groups; and administered dexamethasone s.i.d., at either 0.05 mg/kg bwt i.v. or per os, or 0.02 mg/kg bwt alone or combined with theophylline at 5 mg/kg bwt per os b.i.d. A fifth group was treated with theophylline alone at the above dosage. Lung function was evaluated prior to drug administration and then 3 and 7 days later. RESULTS: Oral administration of dexamethasone alone or combined with theophylline failed to improve lung function significantly in RAO affected horses. Theophylline alone also failed to improve lung function in all treated horses. Conversely, dexamethasone administration at 0.05 mg/kg bwt i.v. resulted in a significant improvement in lung function starting on Day 3. CONCLUSIONS AND POTENTIAL RELEVANCE: Oral theophylline for 7 days did not improve the effects of a low dose of dexamethasone for the treatment of horses with RAO.     

Antioxidant and inflammatory responses of healthy horses and horses affected by recurrent airway obstruction to inhaled ozone

REASONS FOR PERFORMING STUDY: Inhaled ozone can induce oxidative injury and airway inflammation. Horses affected by recurrent airway obstruction (RAO) have a decreased pulmonary antioxidant capacity, which may render them more susceptible to oxidative challenge. It is currently unknown whether RAO-affected horses are more susceptible to oxidative stress than those unaffected by RAO. OBJECTIVES: To determine whether ozone exposure induces greater oxidative stress and airway inflammation in RAO-affected horses in remission than in healthy horses. METHODS: Seven healthy control horses and 7 RAO-affected horses were exposed to 0.8 ppm ozone for 2 h at rest. RESULTS: At baseline, bronchoalveolar lavage fluid (BALF) ascorbic acid concentrations were lower in RAO-affected horses than healthy controls. Ozone appeared to preferentially oxidise glutathione rather than ascorbic acid 6 h after exposure. Individual healthy and RAO-affected horses demonstrated oxidation of BALF glutathione after ozone exposure. Overall, RAO-affected horses did not demonstrate increased oxidative stress following ozone exposure, compared with healthy horses. Ozone did not induce significant airway inflammation in either group. CONCLUSIONS: RAO-affected horses in remission are not more sensitive to ozone despite a decreased pulmonary antioxidant capacity. Sensitivity to ozone appears to be independent of initial pulmonary antioxidant status. POTENTIAL RELEVANCE: Horses with high susceptibility to oxidative stress may benefit from antioxidant supplementation.