BackgroundGastroesophageal reflux and microaspiration (MA) of gastric juice are associated with various human respiratory diseases but not in dogs.ObjectiveTo detect the presence of bile acids in bronchoalveolar lavage fluid (BALF) of dogs with various respiratory diseases.AnimalsTwenty‐seven West Highland White Terriers (WHWTs) with canine idiopathic pulmonary fibrosis (CIPF), 11 dogs with bacterial pneumonia (BP), 13 with chronic bronchitis (CB), 9 with eosinophilic bronchopneumopathy (EBP), 19 with laryngeal dysfunction (LD), 8 Irish Wolfhounds (IWHs) with previous BPs, 13 healthy WHWTs, all privately owned dogs, and 6 healthy research colony BeaglesMethodsProspective cross‐sectional observational study with convenience sampling of dogs. Bile acids were measured by mass spectrometry in BALF samples. Total bile acid (TBA) concentration was calculated as a sum of 17 different bile acids.ResultsConcentrations of TBA were above the limit of quantification in 78% of CIPF, 45% of BP, 62% of CB, 44% of EBP, 68% of LD, and 13% of IWH dogs. In healthy dogs, bile acids were detected less commonly in Beagles (0/6) than in healthy WHWTs (10/13). Concentrations of TBA were significantly higher in CIPF (median 0.013 μM, range not quantifiable [n.q.]‐0.14 μM, P < .001), healthy WHWTs (0.0052 μM, n.q.‐1.2 μM, P = .003), LD (0.010 μM, n.q.‐2.3 μM, P = .015), and CB (0.0078 μM, n.q.‐0.073 μM, P = .018) groups compared to Beagles (0 μM, n.q.).Conclusion and Clinical ImportanceThese results suggest that MA occurs in various respiratory diseases of dogs and also in healthy WHWTs.