Prebiotics are known as components of intestinal microbiota that can improve and maintain human health status by stimulating the growth and activity of the intestinal tract as a method of controlling the intestinal environment. In this study, we examined whether 2’-fucosyllactose (FL) could affect intestinal microbial population and bowel activity. Water content and frequency of mouse feces were increased in the 2’-FL treated group at a high concentration (1,000 mg/kg), with brightness of the color enhanced and physical properties diluted. In addition, intestinal microbial analysis showed that harmful bacteria Clostridium and Staphylococcus strains were decreased and beneficial bacteria such as Lactobacillus strains were markedly increased in the group treated with a high concentration of 2’-FL compared to those in the control group. These findings suggest that administration of 2’-FL can maintain healthy bowel activity by reducing harmful bacteria population and improving diluted physical properties.