Cystic liver disease (CLD), presenting with solitary or multiple cysts in the liver, is a common diagnosis today, primarily due to the frequent application of modern radiological methods. There is a wide range of possible causes. CLD of infective origin is usually caused by an echinococcal species. During the past three decades a number of cystic echinococcosis (CE) control programmes have led to a significant decrease in the incidence of human hydatidosis in some endemic areas. The aim of this study was to determine the seroprevalence of E. granulosus infection in Croatian patients with CLD. A total of 540 serum samples from patients with hepatic cysts detected by imaging methods were screened for the presence of E. granulosus IgG antibodies using semiquantitative enzyme-linked immunosorbent assay. The Western blot technique was used as a confirmatory test for the CE diagnosis. The overall E. granulosus seroprevalence rate in patients with CLD was 3.9%. There was no significant difference in seroprevalence rate between male and female patients (P = 0.541). According to age groups, there was a significant difference in seropositivity among age groups (P = 0.002). The highest seroprevalence rate was detected in the youngest age group (up to 18 years), both in males and females (20% and 13%, respectively). This study indicates that CE still represents a public health problem in Croatia. Preventive measures should be used to control Echinococcus infections, including avoidance of contact with infected dogs, egg-contaminated soil or plants; control and treatment of dogs with antihelmintics; hand washing, improved sanitation and health education.