An 8-yr-old intact female African lion (Panthera leo) presented with a 3-day history of lethargy, anorexia, and vomiting. Hematologic and biochemical abnormalities included a leukocytosis, 41,700/μl (4,700–15,300) with a neutrophilia (37,530/μl; 2,000–9,200) and a left shift (1,250/μl bands; 0–300), and mild hypokalemia of 2.1 mEq/L (2.8–4.8). Abdominal radiographs revealed evidence of intestinal ileus, peritonitis, and the presence of effusion. An exploratory laparotomy was performed, and septic peritonitis due to a pyometra was diagnosed. The lion was treated with an ovariohysterectomy, abdominal lavage, fluid therapy, and a subcutaneous injection of cefovecin. The lion recovered, and clinical signs associated with septic peritonitis resolved within 36 hr. It was returned to conspecifics 3 wk later. Three months postoperatively, the lion showed no residual signs of septic peritonitis.