INTRODUCTION:Gallstone disease is caused by multiple pathogenic factors and is common worldwide. Most studies have focused on the significance of the biliary microbiome in gallstone pathogenesis. Areas covered: In this study, the epidemiology of gallstone diseases and the existence, composition, origin, and mechanisms of the biliary microbiota were reviewed. Mechanisms involved in promoting the formation of different types of gallstones were also emphasized. The antibiotic susceptibility of the biliary microbiota is briefly discussed because it may guide clinical strategies. Expert commentary: The biliary microbiome facilitates the formation of brown pigment stones. Although glycoprotein (mucin) may be pivotal for many promoting substances to coagulate and integrate relevant components, new mechanisms involving prostaglandins, oxysterols, oxygen free radicals, and lipopolysaccharides have been discovered. Furthermore, specific bacterial species such as Helicobacter and Salmonella are involved in the pathogenesis of cholesterol gallstones. Recently, metabolomics of the biliary microbiome has been used to determine the detailed mechanisms that promote gallstone formation. Previously, the bacterial effects involved in the pathogenesis of brown pigment stones have not been analyzed in detail. Whether the administration of antibiotics is related to prophylaxis for gallstone formation and gallstone-associated infections remains unclear.