Gallbladder stasis and gallstone formation are well-known complications of both fasting-associated total parenteral nutrition (TPN) and long-term treatment with octreotide. Additive noxious effects to hepatobiliary function may develop when both therapies are given together as treatment of enteric fistulae. The aim of this study was to assess the risk of gallstone formation during treatment with TPN and octreotide separately and in combination. We studied four groups of New Zealand rabbits (n = 32) during a 2-week intervention period: 14 chow-fed controls; 6 fasted and TPN-fed; 6 chow-fed and administered octreotide, and 6 fasted and treated with both TPN and octreotide. After treatment, the bile duct was cannulated and the gallbladder and liver tissue were obtained for histological analysis. Hepatic and gallbladder bile were retrieved for microscopic examination and measurement of biliary lipids, bilirubin, calcium, total protein, and cholesterol nucleation time. The chemical composition of gallstones was also analyzed. The results of the study suggest that simultaneous administration of these two therapies in rabbits has additive effects on gallbladder stasis and bile lithogenicity. The administration of both treatments can play an important role in the formation of calcium palmitate gallstones in these animals.