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.     

Clinical signs, evaluation of bronchoalveolar lavage fluid, and assessment of pulmonary function in horses with inflammatory respiratory disease

OBJECTIVE: To evaluate the association among clinical signs, results of cytologic evaluation of bronchoalveolar lavage (BAL) fluid, and measures of pulmonary function in horses with inflammatory respiratory disease. ANIMALS: 9 healthy horses, 5 horses with inflammatory airway disease (IAD), and 9 horses with chronic obstructive pulmonary disease (COPD). PROCEDURES: Clinical examination, lung function tests, and BAL were performed on each horse. RESULTS: Standard lung mechanics of horses with exacerbated COPD differed significantly from those of healthy horses; however, there were few differences among horses with IAD, horses with COPD during remission, and healthy horses. Most variables for forced expiration (FE) in horses with COPD or IAD differed significantly from those for healthy horses. Results of clinical examination had low to moderate sensitivity and predictive values for a diagnosis of COPD (range, 67 to 80%). Results of FE tests had high sensitivity, specificity, and predictive values for a diagnosis of COPD (79 to 100%), and results of standard lung mechanics tests had low sensitivity and predictive values (22 to 69%). Percentage of neutrophils in BAL fluid was highly sensitive (100%) but moderately specific (64%) for a diagnosis of COPD. CONCLUSIONS AND CLINICAL RELEVANCE: Clinical examination is moderately accurate for establishing a diagnosis of COPD. Forced expiration tests can specifically detect early signs of airway obstruction in horses with COPD and IAD that may otherwise be inapparent. Cytologic evaluation of BAL fluid allows early detection of inflammatory respiratory disease, but it is not specific for COPD.     

Histamine bronchoprovocation does not affect bronchoalveolar lavage fluid cytology, gene expression and protein concentrations of IL-4, IL-8 and IFN-gamma

In diagnosing inflammatory airway disease (IAD) in performance horses, a histamine bronchoprovocation (HBP) test is often performed. In previously published studies, HBP is usually undertaken prior to cytological examination of the bronchoalveolar lavage (BAL) cells. The purpose of this study was to determine if HBP alters (1) the total nucleated cell numbers and distribution in BAL fluid (BALF) and (2) the mRNA and protein concentrations of selected cytokines in BAL cells and BALF, respectively. BALF was initially collected endoscopically from the right middle or diaphragmatic lung lobe in eight healthy young Standardbred horses. Five to six days later, HBP was performed by aerosolization of histamine (8mg) over a 2min period. BALF was again collected within 2-4h of the HBP from the left middle or diaphragmatic lung lobe. In both samples, total and differential WBC counts were obtained. The gene expressions of interleukin-4 (IL-4), IL-8, interferon-gamma (IFN-gamma) and beta-actin in BAL cells were measured using real-time RT-PCR. The cytokine protein concentrations were measured in the BALF using ELISA. HBP was not associated with either a change in the total BAL cell number or in the distribution of the BAL cells. BAL cell expression of IL-4, IL-8 and IFN-gamma, detected in all samples with the exception of IL-4 in one horse (post-HBP), was not altered as a result of HBP. HBP was not associated with a significant change in IL-8 or IFN-gamma concentrations in the BALF. IL-4 protein was undetectable in BALF either prior to or following HBP. We conclude that HBP can precede BALF collection performed within 2-4h of the former without affecting selected parameters analysed in the BAL cells or BALF.     

Comparison of transtracheal aspirate and bronchoalveolar lavage cytology in 50 horses with chronic lung disease

Comparisons were made between transtracheal aspirate (TTA) and bronchoalveolar lavage (BAL) cytology obtained from 50 horses with chronic lung disease and from 10 control horses. There was no significant correlation between the TTA cytology and the BAL cytology, suggesting that the cell population in the trachea is not representative of the cell population in the lower airways. In control horses the range of differential cell counts obtained from TTA fluid was remarkably large, whereas the variability in cell populations observed in BAL fluid was smaller. In the principal horses the total and differential cell counts of the TTA and BAL fluids were within the 95 per cent confidence interval in 38 and 24 per cent of cases, respectively; and an increase in percentage neutrophils was most common. It was concluded that BAL may be a useful diagnostic aid when evaluating horses with chronic lung disease, but that the clinical usefulness of cytological evaluations of TTA fluid may be limited in these cases.     

Airway inflammation and mucus in two age groups of asymptomatic well-performing sport horses

REASONS FOR PERFORMING STUDY: Horses housed in conventional stable environments are exposed to high dust loads. Long-term exposure to inhaled dust may lead to subclinical airway disease. HYPOTHESES: Airway inflammation and mucus accumulation are 1) common in asymptomatic well-performing sport horses, 2) increased in older compared to younger individuals and 3) correlated. METHODS: Mucus quantity and quality (endoscopic scoring) and BALF differential cytology were assessed in 13 younger horses (mean age 5 years) and 13 older horses (mean age 15 years), which had no historical or clinical evidence of lower airway disease, were performing well and were housed permanently in a conventional stable environment. RESULTS: 1) Airway inflammation and/or mucus accumulation were very common. All sport horses showed evidence of subclinical inflammatory airway disease (IAD). 2) Older horses, having been exposed for 10 more years to a conventional stable environment, did not show increased subclinical airway inflammation or mucus accumulation. The only differences between the age groups were increased BALF total and lymphocyte cell counts in the younger horses. 3) Mucus quantity and quality scores were not significantly correlated with each other or with BALF neutrophil percentages and absolute numbers. CONCLUSIONS: Although clinically healthy and performing well, all of the examined horses housed in a conventional stable environment showed evidence of IAD. The degree of IAD was not increased in older horses that had not developed clinical signs. POTENTIAL RELEVANCE: The effect of subclinical degrees of IAD on performance in dressage and show jumping should be investigated further to better understand the clinical significance of IAD in sport horses.     

Exercising Blood Gas Analysis, Dynamic Upper Respiratory Tract Obstruction, and Postexercising Bronchoalveolar Lavage Cytology—A Comparative Study in Poor Performing Horses

Respiratory abnormalities are common causes of decreased performance in horses presumably because of impaired pulmonary gas exchange. The objectives of the present study were to describe respiratory abnormalities in poorly performing horses and to investigate the relationships between dynamic upper respiratory tract (URT) videoendoscopy, postexercising bronchoalveolar lavage (BAL) cytology, and exercising arterial blood gas analysis. Medical records of 93 horses with exercise intolerance, which presented for treadmill evaluation, were reviewed. Relationships between horse demographics, treadmill endoscopic findings, exercising blood gas values, and BAL cytology results were examined. A total of 25 (27%) horses had a URT obstruction and 91 (98%) horses had abnormal BAL cytology; 73 (78%) had evidence of inflammatory airway disease (IAD) and 83 (89%) had exercise-induced pulmonary hemorrhage (EIPH). In all, 39 (42%) horses had abnormal blood gas values. Dynamic URT obstruction was significantly associated with exercising hypoxemia (P = .036). There were no significant relationships between gas exchange and IAD or between EIPH. Out of 24 (26%) horses with combined URT obstruction and abnormal BAL, horses with URT obstruction and EIPH were more likely to be hypoxic during exercise (P = .037). It was concluded that horses with dynamic URT abnormalities are likely to have exercising hypoxemia. Although IAD and EIPH were commonly indentified in poor performers, they were not significantly associated with abnormal exercising blood gas analysis.     

RADIOGRAPHY OF THE EQUINE THORAX: ANATOMY AND TECHNIC

Radiographic evaluation of the entire adult equine thorax in the standing position requires a minimum of four overlapping lateral radiographs. Small horses could often be studied with fewer radiographs. This basic examination provided the optimal radiographic opportunity for detecting lung diseases in the horse. An understanding of normal gross and radiographic anatomy is an absolute necessity if one is to avoid the potential diagnostic pitfalls inherent in radiographic interpretation of the equine thorax. Of equal importance is an appreciation for the technic required to produce thoracic radiographs. The advent of rare-earth intensifying screen and highspeed film systems has enhanced the opportunity for obtaining diagnostic standing lateral equine thoracic radiographs.     

Clinical, radiographic, endoscopic, bronchoalveolar lavage and lung biopsy findings in horses with exercise-induced pulmonary hemorrhage

A clinical study was performed to determine whether clinical, endoscopic, radiographic, bronchoalveolar lavage (BAL) cytological, and pulmonary biopsy findings could be correlated in horses with exercise-induced pulmonary hemorrhage (EIPH) compared with controls. Racing standardbred horses were selected as either EIPH (n = 10) or control (n = 10), based on repeated postexertional endoscopy of the lower airways. Complete physical and respiratory examinations were performed and blood samples were submitted for arterial blood gas analysis, hematologic study, and fibrinogen determination. Bilateral chest radiographs were taken with the horse standing, and a BAL sample was obtained for cytological examination. Lung was biopsied transcutaneously. Weighted scores were calculated for clinical, radiographic, and pulmonary biopsy findings. The conclusion was that only routine physical examination may help the clinician when EIPH is suspected in horses, especially when there are abnormal findings on percussion of the caudodorsal areas of the chest.     

Bronchoalveolar lavage fluid in Standardbred racehorses: Influence of unilateral/bilateral profiles and cut-off values on lower airway disease diagnosis

The aim of this study was to determine whether the lung side being sampled would significantly influence bronchoalveolar lavage (BAL) cytological profiles and subsequent diagnosis in Standardbred racehorses. One hundred and thirty-eight French Trotters in active training and racing were included in a prospective observational study. BAL was performed using videoendoscopy in both right and left lungs during summer meetings in 2011 (64 horses) and 2012 (74 horses). Cytological data performed 24 h later from right and left lungs were compared and specifically used to classify horses as affected with exercise-induced pulmonary haemorrhage (EIPH), inflammatory airway disease (IAD), or were ‘controls’. For IAD, cytological definition was based on two different cut off values.Neutrophil percentages, haemosiderophage percentages and the haemosiderophage/macrophage (H/M) ratios were significantly higher in the right compared to the left lung. Measures of intra-class correlation coefficients revealed a fair agreement between left and right lungs for percentages of mast cells, eosinophils, and for the H/M ratio, and a moderate agreement for neutrophil percentages. Fair to moderate agreements were observed between left and right lungs for the diagnosis of IAD and/or EIPH based on kappa coefficients. When sampling one lung only, the risk of incorrectly classifying a horse as a ‘control’ increased with the use of the restraint cut-off values for IAD. As BAL from one lung is not representative of the other lung in the same horse, both lungs should be sampled for a better assessment of lung cellularity and for a precise diagnosis of lower airway diseases.     

Comparison of bronchoalveolar lavage and respiratory secretion cytology in horses with histologically diagnosed pulmonary disease

Equine bronchoalveolar lavage (BAL) fluid collected from 70 horses and respiratory secretions (RS) obtained from 61 of these horses were evaluated cytologically and grouped according to the histological diagnosis of the lungs from which they were obtained. The histological categories included: normal lung (8 horses); pulmonary eosinophilic infiltration (9 horses); interstitial pneumonia (5 horses); pulmonary hemorrhage (5 horses); and mild (12 horses), moderate (7 horses) and severe (24 horses) chronic small airway disease. In horses with pulmonary disease, all BAL samples and all but one RS sample differed cytologically to those obtained from normal horses; however, the type and severity of the pulmonary disease could not always be determined using either BAL or RS cytology. There was a positive association between the percentage of neutrophils in BAL and the neutrophil scores in RS specimens; there was no positive association between other cell types.     

Effects of age and allergen-induced airway inflammation in cats: radiographic and cytologic correlation

Thoracic radiography is an important diagnostic tool for feline respiratory medicine. The aim of this study was (1) to assess age-related changes of thoracic radiographic views in healthy young cats and (2) to test if experimentally-induced bronchial inflammation by inhaling Ascaris suum (AS) allergens leads to radiographic changes after single or repeated exposures. Healthy cats (n=15-30) aged between 6 and 30 months were evaluated. Eight healthy cats and eight AS-sensitised cats, respectively, inhaled sterile saline or allergen. Radiographs were taken 24h before, and 6, 24 and 48 h after the challenge. Bronchoalveolar lavage (BAL) was performed after the last radiographic examination. AS-sensitised cats underwent three further allergen challenges at 3-month intervals. The radiographic evaluation was based on a scoring system considering bronchial, interstitial and alveolar patterns. A significant age-related increase in interstitial and total radiographic score was detected in healthy cats older than 18 months and in healthy cats older than 24 months. Whilst saline inhalation did not affect radiographic scores, a single AS challenge induced significant changes of all scores within 6-24h. A significant positive correlation between radiographic scores and BAL neutrophils and eosinophils was found. Repeated AS challenges did not induce irreversible changes in radiographic scores.     

Comparison of results of thoracic radiography, cytologic evaluation of bronchoalveolar lavage fluid, and histologic evaluation of lung specimens in dogs with respiratory tract disease: 16 cases (1996-2000)

OBJECTIVE: To compare results of thoracic radiography, cytologic evaluation of bronchoalveolar lavage (BAL) fluid, and histologic evaluation of biopsy and necropsy specimens in dogs with respiratory tract disease and to determine whether histologic evaluation provides important diagnostic information not attainable by the other methods. DESIGN: Retrospective study. ANIMALS: 16 dogs. PROCEDURE: BAL fluid was classified as normal, neutrophilic, eosinophilic, mononuclear, mixed, neoplastic, or nondiagnostic. Radiographic abnormalities were classified as interstitial, bronchial, bronchointerstitial, or alveolar. Histologic lesions were classified as inflammatory, fibrotic, or neoplastic, and the predominant site of histologic lesions was classified as the alveoli, interstitium, or airway. RESULTS: The predominant radiographic location of lesions correlated with the histologic location in 8 dogs. Of 11 dogs with histologic evidence of inflammatory disease, 8 had inflammatory BAL fluid. Of the 2 dogs with histologic evidence of neoplasia, 1 had BAL fluid suggestive of neoplasia, and the other had BAL fluid consistent with septic purulent inflammation. Two dogs without any histologic abnormalities had mononuclear or nondiagnostic BAL fluid. Two dogs with histologic evidence of fibrosis had mononuclear or mixed inflammatory BAL fluid. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that although thoracic radiography, cytologic evaluation of BAL fluid, and histologic evaluation of lung specimens are complementary, each method has limitations in regard to how well results reflect the underlying disease process in dogs with respiratory tract disease. Lung biopsy should be considered in cases where results of radiography and cytology are nondiagnostic.     

Cytological Analysis of Bronchoalveolar Lavage Fluid in the Diagnosis of Spontaneous Respiratory Tract Disease in Dogs: A Retrospective Study

Results of cytological analysis of bronchoalveolar lavage (BAD fluid were compared with clinical diagnoses in dogs that presented with signs of respiratory disease to referral hospitals. Of 68 dogs in which a clinical diagnosis was possible, BAL cytological findings were considered definitive for the diagnosis in 17 cases (25%), supportive of the diagnosis in 34 cases (50%), and not helpful in 17 cases (25%). Findings were most often considered supportive of or definitive for the clinical diagnosis in dogs with alveolar or bronchial radiographic patterns, or the presence of pulmonary masses. BAL results among lung lobes differed in 23 of 63 dogs (37%) with diffuse radiographic patterns. Tracheal wash cytology differed from BAL fluid cytology in 45 of 66 dogs (68%). Bronchoalveolar lavage was a clinically useful procedure for the diagnostic evaluation of dogs with signs of respiratory disease.     

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.     

Exercise-induced pulmonary haemorrhage in the horse: results of a detailed clinical, post mortem and imaging study. VI. Radiological/pathological correlations

This study was initiated to determine if the extent and intensity of lung lesions associated with exercise-induced pulmonary haemorrhage (EIPH) in horses could be predicted from thoracic radiographs. Sets of thoracic radiographs from 24 horses with varied histories of EIPH were subjectively coded for radiographic quality, and perceived extent and intensity of diffuse interstitial opacity by three radiologists who had no knowledge of the corresponding autopsy results. Codes assigned from radiographs for the chosen parameters were compared with coded estimates of lung surface staining assigned at post mortem and volume measurements of haemosiderin deposits and bronchial arterial neovascularisation recorded from lung slices in separate studies. The non-parametric Spearman rank correlation test was used to test for statistical significance. All radiographically coded estimates of lesion severity were positively correlated with post mortem measurements of actual lesion involvement, but only the correlation between coded estimates of lesion opacity versus haemosiderin deposits and bronchial artery neovascularisation were statistically significant (P less than 0.05). Correlations between radiographic codes for lesion extent versus haemosiderin deposits and neovascularisation were just beyond the level of significance (P greater than 0.05 less than 0.1). These findings indicate that there are graded, radiographically discernible increases in interstitial opacity related to actual lesion severity. However, under the conditions of the study, accurate prediction of lung pathology in individual cases based on radiographic criteria was precluded by the wide variance of the coded values. The authors believe that with good radiographic technique and careful criteria selection, satisfactory prediction of lesion severity in EIPH cases could be achieved.     

Effect of nutritional antioxidant supplementation on systemic and pulmonary antioxidant status,airway inflammation and lung function in heaves-affected horses

An oxidant/antioxidant imbalance in favour of oxidants has been identified as playing a decisive role in the pathogenesis of chronic inflammatory airway diseases. Nutritional antioxidant supplementation might reduce oxidative damage by enhancement of the antioxidant defence, thereby modulating inflammatory processes. In a placebo-controlled, blind study, it was tested whether a dietary antioxidant supplement administered for 4 weeks would improve lung function and reduce airway inflammation in heaves-affected horses. Eight horses in clinical remission of heaves were investigated at rest and after a standardised exercise test before and after treatment with an antioxidant supplement (consisting of a mixture of natural antioxidants including vitamins E and C and selenium from a variety of sources) or placebo (oatfeed pellets without additive). Pulmonary function and exercise tolerance were monitored; systemic and pulmonary lining fluid uric acid, glutathione and 8-epi-PGF(2alpha) were analysed, and bronchoalveolar lavage (BAL) cytology and inflammatory scoring of the airways were performed. The antioxidant treatment significantly improved exercise tolerance and significantly reduced endoscopic inflammatory score. Plasma uric acid concentrations were significantly reduced, suggesting downregulation of the xanthine-dehydrogenase and xanthine-oxydase pathway. Haemolysate glutathione showed a nonsignificant trend to increase, while plasma 8-epi-PGF(2alpha) remained unchanged. Pulmonary markers and BAL cytology were not significantly affected by antioxidant supplementation. The present study suggests that the antioxidant supplement tested modulated oxidant/antioxidant balance and airway inflammation of heaves-affected horses.     

Thoracic Radiographic Features of Silicosis in 19 Horses

Clinical records and thoracic radiographs of 19 horses with a confirmed pathologic diagnosis of silicosis were reviewed. These horses had histories of varying degrees of chronic weight loss, exercise intolerance, and respiratory distress. At the time of presentation, two horses were asymptomatic. Ten horses were geldings and nine were female. The mean age of the 19 horses was 10.7 +/- 5.5 years. Fourteen horses were identified as being from the Monterey-Carmel Peninsula of midcoastal California. An abnormal, structured interstitial pulmonary pattern was identified on thoracic radiographs in each horse. The interstitial pulmonary changes were classified as miliary (13 horses), reticulonodular (4), or linear interstitial (2), and were best visualized dorsally and caudodorsally. In addition to the abnormal interstitial pulmonary pattern, areas of pulmonary consolidation were evident caudodorsally in seven horses. Other thoracic radiographic features included: hilar lymphadenopathy (4 horses), pleural effusion/thickening (4), cranial mediastinal lymphadenopathy (2), hyperinflation (1), and a discrete pulmonary mass (1). Necropsy findings in eight horses and results of lung biopsies in an additional five horses showed a diffuse, multifocal, granulomatous pneumonia with areas of pulmonary fibrosis. Cellular infiltrates included predominantly macrophages with intracellular and/or extracellular crystalline material, occasional lymphocytes, and giant cells. Similar cellular changes were also identified, during necropsy, in the hilar and tracheobronchial lymph nodes in each of the eight horses, although gross enlargement of the lymph nodes was present in only six horses. The radiographic and pathologic findings of these 19 horses are consistent with chronic or the accelerated forms of silicosis that are recognized in humans.     

Effects of pulmonary abscesses on racing performance of horses treated at referral veterinary medical teaching hospitals: 45 cases (1985-1997)

OBJECTIVE: To determine whether results of physical or radiographic examination or biochemical analyses in adult racehorses with primary lung abscesses were associated with ability to race following treatment. DESIGN: Multiple-center retrospective study. ANIMALS: 25 Standardbreds and 20 Thoroughbreds. PROCEDURE: Medical records of horses with a primary lung abscess that were admitted to any of 4 veterinary teaching hospitals were reviewed. Results of physical examination, laboratory testing, and thoracic radiography were reviewed. Racing performance after treatment was compared with performance before illness and with performance of the general population of racehorses of similar age, sex, and breed. RESULTS: 23 of 25 Standardbreds and 13 of 20 Thoroughbreds raced after diagnosis and treatment of a lung abscess. Most horses had a solitary abscess in the dorsal to caudodorsal lung fields. Results of initial physical examination, biochemical analyses, and culture and identification of the microbial isolate were not associated with whether a horse returned to racing. For horses that had raced prior to the illness, race performance after treatment of the lung abscess was not significantly different from performance before the illness. CONCLUSIONS AND CLINICAL RELEVANCE: On the basis of racing performance in those horses that resumed racing after treatment, long-term residual lung damage did not develop in horses with primary lung abscesses that were treated appropriately. It is not known whether horses that recovered would be more likely to bleed from the site of a prior infection when resuming strenuous exercise and whether lung abscesses contributed to a failure to resume racing.