Gallstone formation is a multifactorial disease, caused by the interaction of genetic and enviromental factors. In order to prevent gallbladder stone disease, it is useful to detect modifable risk factors, which contribute to its development. The aim of this study is to analyze the potential relationship between nutrition and the development of gallstone disease, and to establish the possibility for its prevention. The study examined 114 patients; 55 of them suffered from gallstone disease, while 59 were healthy controls who were age- and sex- matched. Diagnosis of gallbladder stone disease was made by ultrasonography. Diet was established using a 24-hour dietary recall method. In the multivariate model, high energy intake (OR = 9.720, p < 0.001) and overnight fasting period (12 hours and longer) (OR = 4.285, p = 0.005) were the most important predictors of gallstone disease, after adjustment for Body Mass Index. These factors can be altered in order to prevent gallstone disease.