Cysto-biliary fistulas are commonly diagnosed, either before or after surgical intervention, in patients with enlarged or advanced-stage hydatid cysts (HCs). Analysis of cystic contents in these patients shows that diminished scolex vitality is more common in patients with cysto-biliary fistulas. This study aims to demonstrate the scolicidal effects of bile on Echinococcus granulosus (EG) scolices at various bile concentrations and over specific periods. The time- and concentration-based scolicidal potency was analyzed in EG scolex samples obtained from HC patients. The integrity of the cyst wall and the scolicidal effect of the contents were evaluated using a vital staining technique with 0.1% eosin. Bile samples were collected from patients with cholelithiasis who were scheduled for surgery. Scolicidal efficacy of 100% was observed in all samples at the 1/4 bile concentration after 10 min and at the 1/8 concentration after 60 min. At a 1/16 concentration, scolicidal efficacy was 66% at 1 min, 89% at 10 min, 93% at 30 min, and 98% at 60 min. At a 1/32 bile concentration, scolicidal efficacy was 59% at 1 min, 68% at 10 min, 89% at 30 min, and 95% t 60 min. At a 1/64 concentration, the scolicidal efficacy was 53% at 1 min, 58% at 10 min, 77% at 30 min, and 93% at 60 min. After 1 hr, reasonable scolicidal efficacy was determined up to a 1/64 concentration; however, significant decreases in scolicidal efficacy were observed at 1/128 and 1/256 bile concentrations. Theoretically, bile, which is an endogenous secretion, can be used as a suitable and potent scolicidal agent. This study lays the groundwork for future in vivo clinical trials utilizing synthetic bile acids for scolicidal purposes or for studies evaluating the scolicidal effects of bile in patients who develop cysto-biliary fistulas.