Mucus accumulation and neutrophilic inflammation in the airways are hallmarks of heaves. Endoscopically visible mucus accumulations, however, have not been studied during exposure to dusty hay and allergens (ie, environmental challenge). We hypothesized that (1) heaves-affected horses have increased mucus accumulation compared with controls, (2) mucus accumulations increase in heaves-affected horses during environmental challenge, and (3) environmental challenge also induces neutrophilic inflammation and mucus accumulation in control horses. Mucus accumulation was graded endoscopically (mucus grades [MGs] 1-5), and airway inflammation was evaluated by bronchoalveolar lavage fluid (BALF) cytology before (0 hours) and during (6, 24, 48 hours) environmental challenge. Large amounts of mucus (MG 4-5) were specific for heaves-affected horses in this study. Variation among controls was considerable, however, and intermediate grades (MG 2-3) were nonspecific, showing complete overlap between the 2 groups. Median mucus accumulations (25th, 75th percentiles) increased in heaves-affected horses from MG 2.5 (1.5, 3.5) at baseline to MG 3.5 (2.0, 4.0), 4.0 (3.0, 4.0), and 4.0 (4.0, 4.0) at 6, 24, and 48 hours, respectively. MG values did not increase in controls--overall MG 1.0 (1.0, 2.0)--even though controls also showed a moderate increase of BALF neutrophils. Mucus accumulations before and especially after exposure to dust and allergens are increased in heaves-affected horses compared with controls. Healthy controls show considerable variability in mucus accumulation but, despite an influx of neutrophils into the airways, no increase of mucus accumulation after exposure to hay dust.