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

Thirty-nine horses and 3 ponies underwent a thorough respiratory examination and were grouped as follows: healthy (4 horses and 1 pony); mild chronic pulmonary disease (CPD 11 horses); moderate CPD (16 horses and 1 pony); and severe CPD (8 horses and 1 pony). Bronchoalveolar lavage (BAL) fluid collected from all animals and respiratory secretions (RS) obtained from 39 of these animals were evaluated cytologically and the results were compared. It was concluded that cytological examination of either BAL fluid or RS was useful in diagnosing various equine pulmonary diseases. The only advantage that BAL offered over RS sampling was in cases in which there was no RS available in the trachea. In addition, the severity of the CPD did not always correlate with either RS or BAL cytology.     

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.     

Equine bronchoalveolar lavage cytology: survey of Thoroughbred racehorses in training

SUMMARY Sixty-two Thoroughbred horses aged between 1 and 7 years in training in Sydney had bronchoalveolar lavage (BAL) samples collected for cytological examination. All horses, except the yearlings and those with a cough, had raced at the time of the examination and the trainers reported satisfactory performance. Free erythrocytes were found In 73% of samples and haemoslderophages In 90% of the samples, Indicating Immediate or past occurrences of exercise-Induced pulmonary haemorrhage (EIPH). Bronchoalveolar fluid from the yearlings contained significantly less (P / 0.05) erythrocytes and haemosiderophages than samples from horses In other age groups. In the older horses, there was also more haemosiderln within the macrophages. No differences In BAL cytology could be attributed to gender, and there was no relationship between BAL cytology and racing performance. The main cytological findings were (mean ± sd): total nucleated cells - 832 ± 578/μL with the main cell types being: macrophages - 59 ± 10% (haemosiderophages - 20 ± 24%); neutrophlls - 9 ± 6%; lymphocytes - 31 ± 9%. The erythrocyte count was 10.3 ± 17.7% of the total cell count. Horses with chronic coughing had a higher proportion of macrophages and a lower proportion of lymphocytes in the leucocytes obtained from BAL. There was a higher occurrence of EIPH detected In BAL findings than that previously reported when endoscopic examination has been used to diagnose EIPH. The occurrence and severity of EIPH as Indicated by the BAL findings was found to be related to exercise Intensity. The cytological findings were similar to those reported in horses in the northern hemisphere. We conclude that BAL cytology may be useful In the diagnosis of various lower respiratory tract disorders and that exercise-induced pulmonary haemorrhage occurs In virtually all horses In race training.     

Cytological findings in bronchoalveolar lavage fluid from feedlot calves: associations with pulmonary microbial flora.

Samples obtained by bronchoalveolar lavage (BAL) were used to evaluate pulmonary cytology in 59 feedlot calves with clinical signs of respiratory disease (cases) and 60 clinically normal comparison calves (controls). Many calves in both case and control groups had inflammatory changes in the lower respiratory tract, as determined by changes in proportions in the BAL differential cell count. Approximately 35% of cases and 40% of controls showed a normal differential cell count. It therefore appeared that the criteria used to select cases for treatment, which were similar to those often used in the field, were poor predictors of lower respiratory tract disease. A positive association was found between an increased proportion of neutrophils in BAL fluid and isolations of Pasteurella multocida and Mycoplasma bovis from BAL fluid.     

Equine multinodular pulmonary fibrosis: a newly recognized herpesvirus-associated fibrotic lung disease

Pulmonary fibrosis and interstitial lung disease are poorly understood in horses; the causes of such conditions are rarely identified. Equine herpesvirus 5 (EHV-5) is a gamma-herpesvirus of horses that has not been associated with disease in horses. Pathologic and virologic findings from 24 horses with progressive nodular fibrotic lung disease associated with EHV-5 infection are described and compared with 23 age-matched control animals. Gross lesions consisted of multiple nodules of fibrosis throughout the lungs. Histologically, there was marked interstitial fibrosis, often with preservation of an "alveolar-like" architecture, lined by cuboidal epithelial cells. The airways contained primarily neutrophils and macrophages. Rare macrophages contained large eosinophilic intranuclear viral inclusion bodies; similar inclusion bodies were also found cytologically. The inclusions were identified as herpesviral-like particles by transmission electron microscopy in a single horse. In situ hybridization was used to detect EHV-5 nucleic acids within occasional macrophage nuclei. With polymerase chain reaction (PCR), the herpesviral DNA polymerase gene was detected in 19/24 (79.2%) of affected horses and 2/23 (8.7%) of the control horses. Virus genera-specific PCR was used to detect EHV-5 in all of the affected horses and none of the control horses. EHV-2 was detected in 8/24 (33.3%) of affected horses and 1/9 (11.1%) of the control horses. This disease has not been reported before, and the authors propose that based upon the characteristic gross and histologic findings, the disease be known as equine multinodular pulmonary fibrosis. Further, we propose that this newly described disease develops in association with infection by the equine gamma-herpesvirus, EHV-5.     

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.     

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.     

Clinical alterations and mRNA levels of IL-4 and IL-5 in bronchoalveolar cells of horses with transient pulmonary eosinophilia

The aim of this study was to assess clinical signs and altered pulmonary cell expression of cytokines related to eosinophil kinetics in horses with pulmonary eosinophilia. Pulmonary eosinophilia was detected by bronchoalveolar lavage (BAL) in a group of standardbreds in training. Horses had detailed clinical examination, bronchoscopy, endobronchial biopsy and BAL on three occasions at approximately 6 month intervals. During the second sampling period BAL eosinophils were significantly elevated (p>0.010), with five horses having from 5% to 37% eosinophils in BAL. Neither detailed clinical examination parameters nor gene expression of IL-4 and IL-5 mRNA (real-time-PCR) were associated with BAL eosinophilia. Pulmonary eosinophilia abated without treatment apart from deworming. It appears that pronounced lung eosinophilia in horses can be transient, abate without specific treatment, and in this instance, lack correlation to upregulation of expression of either IL-4 or IL-5.     

Intradermal testing of horses with chronic obstructive pulmonary disease and recurrent urticaria.

Six horses with chronic obstructive pulmonary disease (COPD) and 8 horses with recurrent urticaria were skin tested with 67 extracts from 58 allergens, including pollens, epidermals, cultivated farm plants, dusts, molds, and insects. Reactions were evaluated 3 times over a 24-hour period immediately after the injections. Results were compared with those obtained from 11 clinically normal horses. All horses had positive skin test reactions. Significant differences was evident between horses with COPD and clinically normal horses for only 3.0% of the possible extract reactions, and between horses with urticaria and clinically normal horses for only 4.5% of the possible extract reactions. Horses with COPD or urticaria had greater total percentage of allergen extract reactions than did clinically normal horses. Positive reactions were observed at all 3 evaluation periods, and late-onset reactions were not always preceded by positive reaction at earlier periods. All horses with COPD or urticaria had at least 1 skin test reaction that exceeded the mean +/- 2 SD, as calculated for each of the 67 extracts for the group of clinically normal horses.     

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.     

Effects of collecting serial tracheal aspirate and bronchoalveolar lavage samples on the cytological findings of subsequent fluid samples in healthy Standardbred horses

Objective To evaluate the effect of collecting serial tracheal aspirate (TA) and bronchoalveolar lavage (BAL) samples on the cytological findings of subsequent fluid samples obtained from horses without clinical signs of respiratory disease. Study design Experimental. Study population Six healthy Standardbred horses. Methods Endoscopically‐guided TA samples, and BAL samples collected using the blind field technique were obtained from the six horses on days 1, 2, 3, 4, 5, 12, and 17. On day 17, horses were sampled three times: at baseline and at 2.5 h and 4 h apart. The differential cytology of the fluid samples collected at each time point was expressed as percentages and compared statistically. Results There was a significant increase in neutrophil percentage in the TA samples taken at day 17 (at 2.5 h but not at 4 h apart). There was no significant change in the neutrophil percentages in the TA samples when repeated samples were taken ≥24 h apart. There was no significant change in the neutrophil percentages in the BAL fluid at any collection point. There were inconsistent changes in the percentages of lymphocytes and macrophages in the BAL fluid over time, but these remained within normal reference ranges and were considered clinically insignificant. Conclusions Serial TA and BAL samples can be taken at 24 h intervals without affecting the cytological findings of subsequent fluid samples collected using the techniques described.     

p65 Homodimer activity in distal airway cells determines lung dysfunction in equine heaves

Nuclear factor-κB (NF-κB) activity, which is a key regulator of inflammatory gene expression, is increased in bronchial epithelial cells from horses suffering from heaves (a hypersensitivity-associated inflammatory condition of the lung). To determine whether this increased activity extends to distal airways and to other pulmonary cells, cells recovered by broncho-alveolar lavage (BAL) in healthy and heaves-affected horses were assessed for NF-κB activity. NF-κB activity was much higher in BAL cells from heaves-affected horses, especially during crisis (disease exacerbation), than in cells from healthy horses. Moreover, the level of NF-κB activity found in BAL cells was positively correlated to total lung resistance and to the proportion of neutrophils present in BAL fluid. Finally, prototypical p65–p50 NF-κB heterodimers were absent from BAL cells, which mostly contained p65 homodimers. These results (1) show that increased NF-κB activity is a general feature of heaves lung; (2) demonstrate the importance of p65 homodimers in neutrophilic inflammation; and (3) suggest that the use of specific NF-κB inhibitors could improve lung function in heaves-affected horses.     

Alveolar clearance in horses with chronic obstructive pulmonary disease

OBJECTIVE: To assess sensitivity of scintigraphic alveolar clearance rate as an indicator of alveolar epithelium damage in horses. ANIMALS: 5 healthy horses (group A) and 5 with chronic obstructive pulmonary disease (COPD; group B). PROCEDURE: Horses underwent clearance rate (k [%/min]) determination. Clearance rate of group-B horses was determined after remission of the disease following 2 months at pasture (remission 1), stabling in a controlled environment (remission 2), and during crisis induced by exposure to moldy hay and straw. Methacholine challenge test was performed at each investigation period to determine nonspecific pulmonary airway hyperresponsiveness. Pulmonary function tests (PFT) also were performed, and cell populations in bronchoalveolar lavage (BAL) fluid were determined on another occasion. RESULTS: Group-B horses had significantly faster mean clearance rate during crisis (k = 4.30+/-0.95%/min), compared with that for remission 1(k = 1.98+/-0.55%/min), which did not differ from the rate in group-A horses (k = 1.95+/-0.33%/min). Despite lack of clinical signs of COPD during remission when stabled in a controlled environment, an intermediate value was found (k = 3.20+/-0.72%/min). CONCLUSIONS: This technique allowed grading of lung damage induced by COPD, whereas use of PFT and determination of BAL fluid cell populations failed to differentiate between remission 1 and remission 2. CLINICAL RELEVANCE: Determination of alveolar clearance rate by use of scintigraphy is a sensitive indicator of lung damage. A modified clearance rate was found despite the lack of clinical and functional changes.     

Comparison of tracheal aspirates and bronchoalveolar lavage in racehorses. 2. Evaluation of the diagnostic significance of neutrophil percentage

OBJECTIVE: To determine whether diagnosis of airway inflammation, using cut-off percentages for neutrophils, differs when based on samples from tracheal aspirate (TA) and bronchoalveolar lavage (BAL) collected concomitantly from the same racehorse. DESIGN: Retrospective case series of 48 young Thoroughbred and Standardbred racehorses in race training, but showing poor performance. PROCEDURE TA and BAL samples were collected from all horses 1 to 2 h after high-speed treadmill exercise. Aliquots of the retrieved fluid were cytocentrifuged and smears stained with Diff-Quik. The mean percentage of neutrophils was calculated. Diagnostic cut-off points were set at 20% for TA samples and 5% for BAL samples. Agreement in the interpretations between the two techniques was analysed. RESULTS: In 19 of 51 paired samples (37%) there were differences in diagnostic interpretation between TA and BAL samples. Of these, airway inflammation was indicated only by the TA sample in 13 and only by the BAL in 6. CONCLUSION: TA and BAL samples give important information about different regions of the airway, but neither should be used alone for the diagnosis of inflammation of the entire lung. The limitations of these procedures mean that both samples should be collected when it is desired to cytologically evaluate the entire lower airway.     

Pulmonary Eosinophilia Associated with Increased Airway Responsiveness in Young Racing Horses

Horses are known to acquire small airway disease (SAD), an allergen-induced naturally occurring syndrome of reversible obstructive lung disease accompanied by airway hyperresponsiveness and increased inflammatory cell numbers on bronchoalveolar lavage (BAL). This disorder has received scant attention in young racehorses. The purpose of the present report was to examine the effect of BAL eosinophilia in young racehorses on clinical examination, BAL, hematology, airway responsiveness, and on pulmonary function at rest and after a standardized exercise challenge. Five (3 males, 2 females; age 2.6 ± 0.9 years) with a history of respiratory compromise and BAL eosinophil differential count <5% and 6 controls (4 males, 2 females; age 3.5 ± 1.0 years) training and performing to expectation with normal BAL cell differential (eosinophils < 1%) were studied. Respiratory system clinical examination was performed and expressed as a clinical score. Arterial blood gas measurements, CBC, and pulmonary function testing were performed at rest. Pulmonary mechanics measurements were repeated 1 hour and 20 hours after a standardized treadmill exercise challenge. Incremental histamine inhalation challenge was performed and the concentration of histamine effecting a 35% decrease in dynamic compliance (PC35C_Dyn) was determined. Significant differences were noted between and controls with regard to clinical score ( P = .01), blood eosinophils ( P = .04), BAL cell count ( P = .04), BAL macrophage differential ( P = .04), PC35C_Dyn ( P = .008), and tidal volume and respiratory rate at 20 hours following exercise challenge ( P = .05). We conclude that pulmonary eosinophilia and airway hyperresponsiveness are manifest in some young horses without overt airway obstruction at rest. We speculate that these may be early events in the natural progression of heaves.     

Cytokine induction in pulmonary airways of horses with heaves and effect of therapy with inhaled fluticasone propionate

Work in humans and laboratory animals has identified a central role for cytokines and chemokines in development and persistence of lower airway inflammation. The objectives of this study were to determine interleukin (IL)-1 beta, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha induction in bronchoalveolar lavage (BAL) of control horses and horses with heaves both during remission and exacerbation of the disease, and to determine the effect of therapy with inhaled fluticasone propionate on the cytokine profile of horses with heaves. IL-1 beta and TNF-alpha mRNA expression was significantly higher in horses with heaves after exposure to moldy hay compared to either values obtained during clinical remission or to healthy controls. IL-8 mRNA expression and protein concentrations were significantly higher in horses with heaves than in controls. Both IL-4 and IFN-gamma mRNA expression was increased at various times in heaves-susceptible horses compared to controls. IL-2, IL-5 and IL-10 mRNA expression was not detected in BAL cells of either group. Therapy with inhaled fluticasone propionate after induction of a severe heaves exacerbation resulted in complete resolution of clinical signs, normalization of pulmonary function tests, and significant decrease in BAL neutrophilia. This was associated with a significant decrease in IL-4 mRNA expression and increase in IFN-gamma/IL-4 ratio in horses with heaves. These results demonstrate the clinical efficacy of inhaled fluticasone propionate for the treatment of heaves and suggest a role for cytokines in the development of lower airway inflammation in heaves-susceptible horses.     

An investigation of the incidence of chronic obstructive pulmonary disease (COPD) in random populations of Swiss horses

The incidence of Chronic Obstructive Pulmonary Disease (COPD) in Switzerland was determined in three groups of horses selected at random. Group A (97 horses) and Group B (93 horses) had no history of respiratory disease, but Group C (113 horses) had a history of lower respiratory tract disease. All horses were examined by auscultation of the respiratory tract under forced breathing, endoscopic examination of the upper and lower respiratory tract, arterial blood gas analysis at rest and cytological examination of respiratory secretions (RS). Fifty-four per cent of the horses in Group A and 54.8 per cent of Group B were diagnosed as suffering clinical COPD and only 12.4 per cent of horses in Group A and 8.6 per cent in Group B had no detectable abnormalities in their lower respiratory tracts. In Group B, 19.4 per cent were considered to be suffering from parasitic pulmonary disease indicated by elevated numbers of eosinophils in their RS. Pharyngeal lymphoid hyperplasia was exhibited in 70.1 per cent of Group A and 74.2 per cent of Group B. Cytological examination of RS was useful to determine if the secretions found in the trachea originated from the upper or lower respiratory tract. Arterial blood gas analysis revealed no significant differences between healthy horses and those with subclinical or mild COPD.     

Serological titers to bovine herpesvirus 1, bovine viral diarrhea virus, parainfluenza 3 virus, bovine respiratory syncytial virus and Pasteurella haemolytica in feedlot calves with respiratory disease: associations with bacteriological and pulmonary cytological variables.

Acute and convalescent serum samples were taken from 59 calves with signs of respiratory disease (cases) and 60 clinically normal animals (controls) during their first month in the feedlot. Sera were analyzed for antibodies to bovine parainfluenza 3 (PI3) virus by hemagglutination inhibition, to bovine viral diarrhea (BVD) virus, bovine respiratory syncytial (BRS) virus and bovine herpesvirus 1 (BHV1) by virus neutralization, and to Pasteurella haemolytica by indirect agglutination (PhIA) and cytotoxin neutralization (PhCN) tests. There was minimal evidence of serological activity to BHV1. Serological activity to the other agents occurred commonly and the prevalence of acute titers and their mean values was similar in case and control groups. Mean convalescent PI3 and P. haemolytica (PhIA) titers were higher in controls than cases (p < 0.01) but, otherwise, convalescent titers did not differ between groups. The incidence of seroconversion was similar in both groups for all agents except for PI3 virus which was more frequent in controls than cases (p < 0.0001). There was a positive association between PhIA and CN seroconversion and isolation of P. haemolytica from bronchoalveolar lavage (BAL) fluid (p < 0.1). The measure of agreement (kappa) between seroconversion with the P. haemolytica PhIA and PhCN tests was 0.51. Bacteriological and cytological evaluations of the respiratory tract were made using BAL. No associations were evident between serological titers and pulmonary cytology. A multivariate logistic analysis was used to evaluate associations between disease status and serological, bacteriological and cytological data. Cases were positively associated with the presence of neutrophils and Pasteurella multocida in BAL fluid and negatively associated with PI3 virus and PhIA seroconversion.     

Pulmonary artery wedge pressure and heart rate measurement during pharmacological stress induction for left cardial function diagnosis in horses with and without heart disease

In 18 horses, the pulmonary artery wedge pressure and the heart rate were measured during pharmacological stress load. 12 horses were healthy (4 trained, 8 untrained) and 6 horses had a heart disease (3 trained, 3 untrained). Pharmacological stress induction was carried out with the sympathomimetic drug dobutamine at a dosage rate of 7.5 microg/kg/min over 10 minutes of infusion. At the fourth minute, the parasympatholytic drug atropine was administered (5 microg/kg bw), and the heart rate and the pulmonary artery wedge pressure were continuously measured over 26 minutes. During sole dobutamine infusion, a significant decrease in heart rate and a significant increase in pulmonary artery wedge pressure were observed. After the application of atropine in the fourth minute, a significant increase in heart rate (from 35.7 +/- 6 up to 106 +/- 38/ min) and in pulmonary artery wedge pressure (from 15.7 +/- 3 up to 24 +/- 8.6 mmHg) were visible in the group of healthy horses. The horses with heart diseases had a significantly higher increase in both parameters (heart rate and pulmonary artery wedge pressure) with a significantly positive correlation (r = 0.7). The heart rate increased in the horses with heart diseases from 35.2 +/- 2,8 beats/min up to 132 +/- 45.7 beats/min and the pulmonary artery wedge pressure increased from 17.3 +/- 3,2 mmHg up to 32.7 +/- 13 mmHg. The cardiac status (healthy or heart disease) as well as the training level of the horses (untrained or trained) had a significant influence on the heart rate and the pulmonary artery wedge pressure. The untrained horses (healthy and heart disease) showed significantly higher values over a longer period of time than did the trained horses with the same cardiac status. Additionally the influence of pharmacological stress induction on echocardiographic parameters was investigated. The left atrial size (p = 0.015) and left ventricular diameter were significanly different in the systole (p = 0.008) and in the diastole (p = 0.001) between healthy horses and horses with heart diseases. All horses showed a positive correlation between the pulmonary artery wedge pressure and the left atrial size (r = 0.8), as well as between the left ventricular systolic (r = 0.6) and the diastolic diameter (r = 0.6). The correlation between the pulmonary artery wedge pressure and the left atrial size was nearly the same in the healthy horses (r = 0.74) and in the horses with heart diseases (r = 0.76). Regarding the training level, all untrained horses had a significantly higher correlation between the pulmonary artery wedge pressure and the left atrial size (r = 0.87) in comparison to the trained horses (r = 0.74). Particularly in the untrained horses with heart diseases, this correlation was remarcable (r = 0.99).