Insect Bite Hypersensitivity in Horses is Associated with Airway Hyperreactivity.

BACKGROUND: Genetic and epidemiologic evidence suggests that in horses, as in other species, different manifestations of hypersensitivity may occur together. HYPOTHESIS: Horses affected with insect bite hypersensitivity (IBH) show airway hyperreactivity (AH) to inhaled histamine, even in the absence of overt clinical signs of equine asthma (EA). ANIMALS: Twenty-two healthy controls (group C), 24 horses suffering from IBH alone (group IBH), and 23 horses suffering from IBH and EA (group IBH/EA). METHODS: The clinical histories were assessed using 2 standardized questionnaires, the Horse Owner Assessed Respiratory Signs Index (HOARSI), and IBH scoring. Horses were classified as EA-affected if their HOARSI was >1 and as IBH-affected if IBH score was >0. Confounding disorders were excluded by clinical examination. The arterial partial pressure of oxygen (PaO2 ) was measured and flowmetric plethysmography used to assess airway reactivity to increasing doses of inhaled histamine. RESULTS: The median histamine provocation concentration (PC) when ∆flow values increased by 35% (PC35) was significantly higher in group C (5.94 [1.11-26.33] mg/mL) compared to group IBH (2.95 [0.23-10.13] mg/mL) and group IBH/EA (2.03 [0.43-10.94] mg/mL; P < 0.01). The PC50 and PC75 showed very similar differences between groups. Furthermore, PaO2 was significantly lower in group IBH (84 ± 8 mmHg) and group IBH/EA (78 ± 11 mmHg) compared to group C (89 ± 6 mmHg; P < 0.01). CONCLUSIONS AND CLINICAL IMPORTANCE: IBH is associated with AH and decreased PaO2 , even in the absence of overt respiratory clinical signs.

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(-7)M to 10(-4)M) 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.

Effects of neuraminidase on equine isolated bronchi.

It has been demonstrated in mammals that the airway hyper-responsiveness, which accompanies viral infections, is the result of increased reflex bronchoconstriction due to inhibition of muscarinic prejunctional receptors, which belong to M2 subtypes. Multiple mechanisms account for virus-induced M2 receptor dysfunction. Viral neuraminidase may deglycosylate the M2 receptor, decreasing acetylcholine affinity. Equine influenza remains a common viral respiratory disease of horses worldwide, which results in loss to the equine industry, by decreasing performance, convalescence time and loss of peak performance due to chronic sequelae, such as airway hyper-responsiveness. The purpose of this study was to evaluate the effect of neuraminidase on equine isolated bronchi, assessed in equine bronchial smooth muscle rings, derived from five healthy equine male lungs. A pretreatment with vehicle did not modify contraction induced by electrical field stimulation (EFS) studies at each frequency tested. A pretreatment with pilocarpine (1-100 microM) significantly reduced, while methoctramine (1-100 microM) significantly increased contraction induced by EFS. Finally, neuraminidase (0.5 UI) significantly increased contraction induced by EFS. These results suggest that airway hyper-responsiveness that follows a viral influenza infection might be related to a dysfunction of muscarinic prejunctional receptors.

Altered airway responsiveness in adult sheep born prematurely: effects of allergen exposure.

The aim of this study was to determine the effects of preterm birth per se on airway function in adult sheep. Preterm birth was induced at approximately 0.89 of term. At approximately 1 year of age the authors measured pulmonary resistance (RL) and airway responsiveness before and after house dust mite (HDM) challenge. Mature preterm sheep tended to have greater baseline RL than controls (P = .12): the smaller preterm sheep showed significantly greater RL than controls following bronchoconstrictor challenge. Preterm animals tended to have greater baseline total blood leukocyte count (P = .06). It was concluded that preterm sheep, especially with low postnatal growth, have greater airway responsiveness to bronchoconstrictor and higher baseline RL.

Endothelin receptor alterations in equine airway hyperreactivity.

The purpose of this study was to evaluate the role of endothelin-1 (ET-1) and its receptors in the airway hyperreactivity of horses with obstructive pulmonary disease associated with summer pasture (SPAOPD). The right diaphragmatic lobe of the lung of 8 clinically healthy (unaffected) and 8 SPAOPD-affected horses was collected immediately after euthanasia. Bronchial rings (4 mm wide) were prepared and mounted in organ baths and attached to force transducers interfaced with a polygraph. Four rings were used to study each ET-1 receptor; 1 ring served as the control, and the other 3 were incubated with 10(-9), 10(-7), or 10(-5) M of either BQ-123, an ET(A)-receptor antagonist, or IRL-1038, an ET(B)-receptor antagonist. Cumulative concentrations (10(-8.5) to 10(-6) M) of ET-1 were applied to all rings. Using pooled pulmonary tissue from different regions of the lung, we performed a reverse-transcription polymerase chain reaction (RT-PCR) to determine ET(B)-receptor gene expression. Although ET-1 caused concentration-dependent bronchial ring contraction in both groups of horses, the rings of SPAOPD-affected horses had significantly greater contraction than the rings of unaffected horses. Whereas ET(A)-receptor blockade significantly increased the response to ET-1 in unaffected horses, ET(B)-receptor blockade significantly decreased the response in affected horses. The pA2 values showed a nonsignificant decrease in ET(A)-receptor affinity and a significant increase in ET(B)-receptor affinity in affected horses compared with unaffected horses. The ET(B)-receptor mRNA expression of the pooled pulmonary tissue showed a nonsignificant increase in affected horses compared with unaffected horses. The airway hyperreactivity to ET-1 observed in the bronchial rings from the affected horses appears to be due in part to activation of pulmonary ET(B) receptors, which appear to be inactive in unaffected horses.

Whole body barometric plethysmography: a screening method to investigate airway reactivity and acute lung injuries in freely moving pigs.

The aim of this work was to investigate whether the Penh index, measured using whole body barometric plethysmography, can be used as a screening parameter to evaluate the airway reactivity and the intensity of the pulmonary response to endotoxins. Penh was firstly recorded in non-sedated freely moving piglets exposed (1) to a nebulized acetylcholine (Ach) pre-treated or not with clenbuterol, or (2) to endotoxin challenge. To measure Penh simultaneously with total pulmonary resistance (R(L)), dynamic compliance (C(dyn)) and intrapleural pressure changes (Max Delta Ppl), an oesophageal balloon catheter technique was used and the piglets were anaesthetised. The recordings were performed during (1) an intravenous metacholine (Mch) challenge and (2) in endotoxin-exposed animals. In freely moving animals, Ach induced a significant dose-dependent increase in Penh, which was significantly blocked by clenbuterol. Endotoxin instillation also resulted in a significant rise in Penh while the corresponding response measured under anaesthesia was significantly and positively correlated with R(L) and Max Delta Ppl. Similar results were obtained during Mch challenge but the Penh was negatively correlated with C(dyn). We conclude that Penh could be used in freely moving piglets as a screening index for airway reactivity and pulmonary functional changes in cholinergic and endotoxin challenges.

Allergen challenge alters lymphocyte phosphodiesterase activity in horses with heaves.

Heaves is an allergic airway disease in horses characterised by reversible airway obstruction, bronchial hyperresponsiveness and airway inflammation associated with a Th(2) response. Cyclic nucleotide-dependent signalling pathways can regulate lymphocyte function. In this study, we examined lymphocyte PDE activity comparing horses with heaves to healthy control animals. Total PDE activity and the effects of isoenzyme selective inhibitors were measured before, 5 and 24 h after the start of a 7 h allergen challenge. Allergen challenge had no effect on either total cAMP PDE activity or its inhibition by the PDE4 selective inhibitor, rolipram, and the non-selective PDE inhibitor, theophylline. In contrast, the PDE3 selective inhibitor, quazinone, caused significantly greater inhibition of cAMP PDE activity before challenge in the heaves susceptible group. Additionally, total cGMP PDE activity was significantly lower 24 h after the start of challenge in the heaves affected group (11+/-2 and 21+/-3 pmol/min/mg for heaves and control animals, respectively) and the PDE5 selective inhibitor, zaprinast, caused significantly less inhibition in the heaves group at this time point. The functional significance of these findings was explored by examining the effect of PDE3, PDE4 and PDE5 selective inhibitors on mitogen-induced mononuclear cell proliferation before and 24 h after the start of allergen challenge. Proliferation decreased after challenge in the heaves group (stimulation index=328+/-110 and 200+/-72 before and after challenge, respectively) whilst remaining constant in the control group (stimulation index=161+/-13 and 183+/-45 before and after challenge, respectively). However, all three PDE inhibitors caused a similar amount of inhibition at each time point and the effect of a combination of a PDE3 and a PDE5 inhibitor was simply additive in both groups. These results suggest differences in the control of lymphocyte PDE activity in horses with heaves.

Comparative responses of bronchial rings to mediators of airway hyperreactivity in healthy horses and those affected with summer pasture-associated obstructive pulmonary disease.

OBJECTIVE: To compare responses of bronchial rings obtained from healthy horses and horses affected with summer pasture-associated obstructive pulmonary disease (SPAOPD) to selected mediators of airway hyperreactivity in vitro. SAMPLE POPULATION: Bronchial rings from 6 healthy horses and 6 horses affected with SPAOPD. PROCEDURE: Bronchial rings obtained from each group of horses were mounted in organ baths and attached to force transducers interfaced with a polygraph. After applying 2g of tension, each ring was allowed to equilibrate for 45 minutes in Tyrode's solution at 37 C. Cumulative concentration-response relationships to graded concentrations of selected mediators (10(-8) to 10(-4) M) were determined and analyzed for significance at each concentration. RESULTS: Acetylcholine, histamine, 5-hydroxytryptamine, and leukotriene D4 induced concentration-dependent contractile responses in bronchial rings. Prostaglandin F2alpha induced weak and inconsistent contractile responses. The other 2 agents, norepinephrine and substance P, did not induce concentration-dependent responses. Considering the overall group-drug effect, acetylcholine, histamine, 5-hydroxytryptamine, and leukotriene D4 were effective in inducing consistent concentration-dependent contractile responses in both groups. Only 5-hydroxytryptamine and histamine induced significant responses in contractility between groups. The response of bronchial rings from horses with SPAOPD to 5-hydroxytryptamine was significantly greater than those from control horses, whereas the response to histamine was significantly lower. Significant responses were evident at concentrations ranging from 10(-6) to 10(-4) M for both drugs. CONCLUSIONS AND CLINICAL RELEVANCE: Because the airways of horses with SPAOPD had increased responsiveness to 5-hydroxytryptamine in vitro, treatment modalities using 5-hydroxytryptamine antagonists should be investigated to address this phenomenon.

Comparison of forced oscillation with the conventional method for histamine bronchoprovocation testing in horses.

OBJECTIVE: To compare response of horses to histamine bronchoprovocation (HBP), using total respiratory resistance (Rrs) measured by forced oscillatory mechanics (FOM) with dynamic compliance (Cdyn) and pulmonary resistance (RL) measured by the esophageal balloon method. ANIMALS: 10 horses with various degrees of airway reactivity. PROCEDURE: The 2 methods for measuring airway responses to HBP were performed on separate days. Endpoints compared were increase of 150 or 200% of baseline Rrs (PC150Rrs, PC200Rrs) and to 150% of baseline RL (PC150RL) or decrease to 65% of baseline Cdyn (PC65Cdyn). Frequency dependence of respiratory mechanics, using FOM, was evaluated, using the slope of Rrs over 1 to 3 Hz and the ratios of Rrs at 1 Hz to Rrs at 2 and 3 Hz (Rrs1Hz/Rrs2Hz, Rrs1Hz/Rrs3Hz) and of Rrs at 2 Hz to Rrs at 3 Hz (Rrs2Hz/Rrs3Hz). Effect of histamine on frequency dependence was assessed. RESULTS: Correlation with PC65Cdyn was high for PC150Rrs (rs = 0.93) and PC200Rrs (rs = 0.96). Correlation between PC65Cdyn and PC150RL was weakly positive (rs = 0.61). The slope of Rrs over 1 to 3 Hz changed significantly between baseline (-0.07+/-0.09 cm H2O/L/s/Hz) and final histamine dose (-0.28+/-0.10 cm H2O/L/s/Hz). The Rrs1Hz/Rrs3Hz and Rrs2Hz/Rrs3Hz differed significantly (P < 0.05 between baseline [1.27+/-0.36 and 0.96+/-0.11, respectively] and final histamine dose [1.62+/-0.37 and 1.09+/-0.14, respectively]). CONCLUSIONS: Correlation between histamine-induced changes in respiratory mechanics, as measured by FOM and the pneumotachograph-esophageal balloon method, was good. The FOM results indicated frequency dependence of respiratory mechanics during HBP. CLINICAL RELEVANCE: A noninvasive method of measuring airway reactivity will facilitate routine evaluation of horses with suspected small airway disease and may be suitable for field studies.

Airway responsiveness to acetylcholine in congenitally bronchial-hypersensitive (BHS) and bronchial-hyposensitive (BHR) guinea pigs in vivo and in vitro.

The characteristics of airway responsiveness to acetylcholine (ACh) in congenitally bronchial-hypersensitive (BHS) and bronchial-hyposensitive (BHR) guinea pigs were clarified in vivo and in vitro. We measured the change in ventilatory mechanics in response to ACh inhalation by means of the bodyplethysmograph and the contractile responses of isolated trachea to ACh and carbachol (CCh). Further, muscarinic receptor subtypes involved these responses were identified. The basal values for ventilatory mechanics in BHS were not significantly different from those in BHR. Respiratory resistance to ACh was progressively increased in a time- and dose-dependent manner in BHS. The contractile responses of tracheal smooth muscle to ACh in BHS were significantly greater than those in BHR, but CCh-induced responses in BHS and BHR were similar. ACh- and CCh-induced contractions were mediated via M3 receptors. These results suggested that the falling-down of BHS in response to ACh inhalation was caused by the strong constriction of the airway and the reduction in ventilation. Moreover, the airway hyperresponsiveness to ACh in BHS might be partly dependent on the change in acetylcholinesterase activity.

Association between bronchoalveolar lavage cytologic features and airway reactivity in horses with a history of exercise intolerance.

OBJECTIVE: To correlate indices of airway reactivity to bronchoalveolar lavage (BAL) fluid cytologic features in horses with a recent decline in exercise tolerance. ANIMALS: 20 actively working horses from 2 to 24 years old. PROCEDURE: Bronchoalveolar lavage fluid samples were obtained and analyzed. Forced oscillatory mechanics (1-7 Hz) technique was used for measurements of total respiratory system resistance (RRS), compliance (CRS), and resonant frequency (fres). Changes in RRS (1 Hz) during histamine challenge were used to generate histamine dose-response curves, from which the provocative concentrations that evoked a 75 or 100% increase in baseline RRS (PCRRS75 and PCRRS 100, respectively) were determined. Age, sex, baseline lung mechanics, and BAL cytologic findings were correlated with PCRRS75 and PCRRS100. RESULTS: No horse of the study had clinical signs or history of obstructive pulmonary disease or increased percentage (> 7%) of neutrophils in bronchoalveolar lavage fluid samples. Mean (+/- SEM) RRS, CRS, and fres were 0.67 +/- 0.06 cm of H2O/L/s, 0.52 +/- 0.04 L/cm H2O, and 2.46 +/- 0.02 Hz, respectively. There was no correlation between age or sex, and RRS, CRS, fres, PCRRS75, or PCRRS100. There was a significant correlation (rs = -0.78, P < 0.001) between percentage of BAL fluid mast cells and PCRRS75 or PCRRS100, but correlation with other cell types and indices of airway reactivity were not observed. CONCLUSION: The strong association between mast cell percentage in BAL fluid and airway reactivity in this group suggests that mast cell products may contribute to bronchospasm, airway wall thickening, and/or loss of elastic recoil, which underlie airway hyperreactivity. Alternatively, mast cells may contribute to nonspecific airway reactivity in horses through unknown mechanisms.

Small airway disease as a vanguard for chronic obstructive pulmonary disease.

Equine allergic small airway disease is a highly prevalent respiratory condition among the stabled horse population. With the assistance of new diagnostic tools such as bronchoalveolar lavage, the condition can be recognized in young performing horses. The pathophysiological and clinical features resemble an earlier stage of chronic obstructive pulmonary disease as determined by the appearance of specific inflammatory cells. Although environmental management is paramount in controlling the disease, proper selective therapeutic regimens are as important to reduce the concurrent inflammation and to reduce exacerbations of the disease.

Allergic respiratory disease.

Allergic rhinitis and extrinsic allergic alveolitis are the most common allergic disorders of the bovine respiratory system. Environmental and management factors play significant roles in the pathogenesis of these disorders. When compared to infectious or toxic respiratory disease, allergic respiratory disease is relatively rare and of far less economic importance in North American cattle; however, the environmental and management conditions conducive to these diseases exist in many regions. Therefore, familiarity with the clinical and epidemiologic features of these unique diseases will aid the veterinarian in establishing an accurate diagnosis. Signs of respiratory dysfunction are common to anaphylactic and anaphylactoid reactions. Early recognition of these adverse reactions will provide the practitioner with the greatest chance of successful treatment.

Respiratory diagnostic pathology.

Effective treatment and control of bovine respiratory disease is dependent upon an accurate diagnosis. This article discusses the approach to diagnosis of bovine respiratory disease from the perspective of respiratory pathology. Topics covered include necropsy examination of the respiratory system, sample collection and submission, and the gross, and histopathologic lesions of the upper and lower bovine respiratory system.

Duration of antigen-induced hyperresponsiveness in horses with allergic respiratory disease and possible links with early airway obstruction.

Antigen-induced airway hyperresponsiveness in allergic horses has previously been demonstrated when clinical signs of acute airway obstruction were apparent, as a consequence of exposure of animals to hay and straw for variable periods of time, and repeat measurements of hyperresponsiveness have been made no earlier than 1 week after challenge. In the present study airway responsiveness to methacholine has been measured in normal horses and allergic horses in clinical remission before and 24, 48 and 72 h after a hay and straw challenge of fixed, short, duration (7 h). Correlations between early increases in interpleural pressure and hyperresponsiveness have also been investigated. As in other studies, the mean airway responsiveness of groups of normal and allergic horses in clinical remission was not significantly different. The responsiveness to methacholine of allergic, but not normal, horses was increased after antigen challenge and was significantly greater than that of normal horses at 48 and 72 h after challenge (log PD8 cm: -0.77 +/- 0.28 vs. 0.27 +/- 0.14 at 48 h and -0.6 +/- 0.25 vs. 044 +/- 01 at 72 h; P < 0.05). There was also a significant correlation between interpleural pressure at the end of the 7-h challenge in allergic horses and the increase in responsiveness to methacholine at 24, 48 and 72 h. These results demonstrate that antigen induces an increase in airway responsiveness in allergic horses that persists for up to 3 days and which may be linked to the initial increase in interpleural pressure.

Oxygen radicals contribute to antigen-induced airway hyperresponsiveness in conscious sheep.

We previously showed that oxygen radicals can induce airway hyperresponsiveness (AHR) in allergic sheep. The purpose of this study was to determine whether antigen challenge results in the generation of free oxygen radicals and if these radicals contribute to antigen-induced AHR. We first determined baseline airway responsiveness in seven Ascaris suum-sensitive sheep by calculating the cumulative provocative concentration of carbachol in breath units (BU; one BU defined as one breath of a 1% wt/vol carbachol solution) that increased specific lung resistance (SRL) 400% over baseline (PC400). On a different day, the sheep underwent inhalation challenge with A. suum antigen, SRL was measured before and immediately after challenge and then hourly for 2 h, at which time SRL had returned to baseline. The postchallenge PC400 was then measured. This procedure was repeated on separate occasions, each at least 14 days apart, except that the sheep were treated with an aerosol of catalase (CAT; 38 mg in 3 ml deionized water), the enzyme that catalyzes the decomposition of hydrogen peroxide (H2O2), at three different times: Trial 1, before antigen and then every 30 min after antigen challenge for 2 h; Trial II, 1 and 2 h after antigen challenge; and Trial III, only at 2 h after antigen challenge. In the control trial, antigen challenge caused a transient (mean +/- SEM) 303 +/- 48% increase in SRL over baseline (p < 0.05), and 2 h later, PC400 was reduced to 11.0 +/- 1.7 BU from a prechallenge value of 24.8 +/- 1.9 BU (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)