Dietary yeast culture supplementation can contribute to the performance and health of sows and piglets, but few studies have focused on the relationships between the effects of yeast culture and gut microbiota. This study investigated the effect of yeast culture (Saccharomyces cerevisiae) supplementation from late gestation to weaning on the reproductive performance of lactating sows and their faecal microbiota. One hundred and six purebred Landrace sows, of parities two to six were selected and randomly assigned to a control (CON) and yeast culture supplementation (YC) groups based on parity and back fat thickness. The YC sows were individually fed with yeast culture at a dose of 24 g/d from day 90 of gestation to parturition and 40 g/d during lactational period. Blood samples were collected from sows on d 110 of gestation and at weaning at day 21 of lactation for plasma hormone and immunoglobulin analysis. Colostrum and milk on day 20 of lactation were collected for composition analysis. Faecal samples from sows on day 110 of gestation and day 20 of lactation were collected for short-chain fatty acid and faecal microbial analysis. Results showed that the farrowing performance of YC sows did not differ significantly from the CON group (P > 0.05). The average daily feed intake by the YC group during the lactation period was significantly increased by 9.98% (P = 0.004), the weaning-to-oestrus interval was shortened by 0.96 d (P = 0.046) and average daily weight gain of piglets increased by 7.14% (P = 0.036) compared with the CON group. Yeast culture supplementation also significantly improved the average daily milk yield in the first week of lactation (P = 0.035), lactose content in colostrum (P = 0.046), protein (P = 0.033) and DM (P < 0.001) content of milk. In the YC group, concentrations of plasma ghrelin (P = 0.02) and IgG (P = 0.015) were increased compared with the CON group, while that of glucagon-like peptide-1 was decreased (P = 0.006) on d 110 of gestation. The 16S rRNA gene sequencing showed that faecal microbiota changed at taxonomic levels with yeast culture addition (P < 0.05). Dietary yeast culture supplementation from late gestation to lactation improved feed intake, immunity status, milk yield, milk quality and faecal microbiota of sows, resulting in the improved growth performance of piglets.