A 12-year-old African lioness from a safari park presented with abdominal discomfort and severe jaundice. Postmortem examination revealed a tortuous gallbladder, divided internally by thin septa into multiple chambers. Microscopically, multiple sacculations separated by septa of loose fibrovascular tissue were seen. These sacculations had intercommunicating openings and pouch-like endings. The gallbladder epithelium showed diffuse coagulative necrosis, but numerous cystic formations were visible. Neither thickening of the gallbladder wall nor inflammatory infiltrates were identified. The lesion was diagnosed as multiseptate gallbladder, a rare congenital malformation described in human beings.