Canine cholecystitis is diagnosed infrequently. Clinical signs, physical examination findings, and clinicopathological abnormalities are nonspecific. Few reports exist of associated ultrasonographic findings which also are nonspecific for the disorder. Ultrasonographic-guided, percutaneous cholecystocentesis has been suggested for diagnostic confirmation. The present report further documents ultrasonographic findings associated with canine acalculous cholecystitis and demonstrates the utility of ultrasonographic-guided, percutaneous cholecystocentesis in confirmation of the diagnosis in a prospectively acquired clinical case series.