Palm kernel expeller (PKE), a by-product of palm oil extraction, contains higher amounts of fiber than corn and soybean meal, but offers low energy density, protein value, and amino acid (AA) composition, limiting its use for swine. Recently however, it was reported that dietary fiber has a positive effect on the gut microbiota of the host, and therefore it is necessary to study the effect of PKE feeding on the intestinal microbiota of swine. In this study, we investigated the effects of supplementation with PKE in lactation diets on the gut microbiota composition of lactating sows and their litters. A total of 12 sows were randomly assigned to two dietary treatment groups in a completely randomized design. The treatments were a diet based on corn-soybean meal (CON) and CON supplemented with 20% of PKE. Sow and piglet fecal samples were collected before farrowing, on days 7 and 28 (weaning) after farrowing, and on days 7 and 28 (weaning) after farrowing, respectively, to verify gut microbiota composition by pyrosequencing analysis. The beta-diversity result showed a significant difference only in weaning-stage piglets, but dietary PKE altered the gut microbiota in sows by increasing the abundance of compared with CON. In piglets, dietary PKE decreased the abundance of opportunistic pathogen and increased the abundance of potentially beneficial bacteria, such as and . Our results can be helpful in developing feeding strategies and support the beneficial effects of dietary PKE to improve the gut health of animals.