This study explored the effect of dietary xylo-oligosaccharide (XOS) supplementation on the gut microbial composition and activity in pigs of different ages. Eighty pigs with an average body weight (BW) of 30 kg were randomly divided into eight groups: A control group, a group that received antibiotic treatment, and six groups fed diets supplemented with 100, 250, and 500 g/t XOS, of which three groups were in the growing period (GP, 30-65 kg BW) and three groups in the growing and fattening period (GFP, 30-100 kg BW). At the end of the experiment, the intestinal contents were sampled for analyses of gut microbiota and bacterial metabolites including short-chain fatty acids (SCFAs) and bioamines. The results showed that 100 g/t XOS supplementation during the GFP significantly reduced the relative abundances of presumably pathogenic bacteria (Proteobacteria and Citrobacter), but enhanced the relative abundances of likely beneficial bacteria (Firmicutes and Lactobacillus). However, XOS supplementation during the GP showed little effect on the gut microbiota when pigs were killed at 100 kg BW. Meanwhile, 100 g/t XOS supplementation during the GFP decreased the level of 1,7-heptane diamine and increased the acetic acid, straight-chain fatty acids, and total SCFAs concentrations in the intestinal contents. Statistical analysis showed that both the dose and exposure time to XOS supplementation affected the microbial communities. In summary, 100 g/t XOS supplementation during the GFP modified the gut microbiota composition and metabolic activity. Possible consequences of such changes for the host are discussed.