Simple SummaryThe equine hindgut is colonized by microorganisms, some of which are involved in fiber digestion and are crucial for the horse’s nutrition and health. These key microorganisms are very sensitive to dietary changes, which have been identified as a risk factor for colics. This study assessed the stressful effect of two consecutive abrupt dietary changes on the diversity, the composition, and the activity of fecal microorganisms focusing on fibrolytic bacteria. Twelve horses were subjected to an abrupt change from forage to a concentrate-rich diet, followed by a second change from a concentrate to forage-rich diet 5 days later. Half of the horses were given a supplement of living bacteria as a probiotic. Two days after the sudden change from forage to concentrate diet, the proportions and types of microorganisms were altered drastically, as was their capacity to degrade fibers. After this dietary stress, it took 3–4 weeks of a high-fiber diet to recover the basal state. Supplementation with probiotics promoted an earlier recovery of fibrolytic bacteria after the dietary stress. Abrupt dietary changes should be limited in horse management to protect the hindgut microorganisms and their capacity to use forage fibers, and consequently to limit the development of colic.In horses, abrupt changes from high-fiber (HF) to high-starch (HS) diets can affect the cecal and colonic microbiota. This study investigated modifications and recovery of fecal microbiota after two consecutive abrupt dietary changes. Twelve horses fed HF for 2 weeks were changed to HS for 5 days then returned to HF for 7 weeks. Six received lactic acid bacteria supplementation. Bacterial population diversity, structure, and activity, especially fibrolysis, were assessed to obtain an overview of alteration in hindgut microbiota. Two days after the abrupt change from HF to HS, the findings in feces were consistent with those previously reported in the cecum and colon, with a decrease in fibrolytic activity and an increase in amylolytic activity. Fecal parameters stabilized at their basal level 3–4 weeks after the return to HF. A bloom of cellulolytic bacteria and lower pH were observed after 1.5 weeks, suggesting a higher level of fiber degradation. In supplemented horses the relative abundance of potentially fibrolytic genera was enhanced 2 days after HS and 2 days to 2–3 weeks after the return to HF. Fecal analysis could be a promising technique for monitoring hindgut microbial variations accompanying dietary changes.