The intestinal mucus layer is shaped and modulated by the microbiota. In a new study, the inner mucus layer of germ-free mice was found to be more easily penetrable to bacteria-sized beads compared with conventionally raised mice. When germ-free mice were inoculated with conventional bacteria, mucus in the small intestine detached 5 weeks after colonization, and the colonic inner mucus required 6 weeks to become impenetrable. The microbiota composition of the small intestine was similar in conventionally raised donor mice and colonized germ-free mice for 3 weeks, with shifts thereafter and normalization after 7 weeks.