This study was conducted to evaluate the effects of yeast culture (YC) supplementation on the growth performance, apparent nutrient digestibility and caecal microflora of broiler chickens. A total of 360 one-day-old Arbor Acres broiler chickens were randomly assigned to six dietary treatments containing 0.2%, 0.4%, 0.6%, 0.8% and 1% YC. The experiment lasted for 42 days. Diet and faecal samples were collected for analysis of dry matter, crude protein, ether extract, calcium and phosphorus. Caecal microbiota on days 21 and 42 were measured using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and real-time PCR. Dietary supplementation with YC did not affect feed intake. On day 42, the 0.8% YC group showed optimal growth and feed efficiency, as well as higher levels of apparent digestibility of ether extract, calcium and phosphorus. On day 21, both 0.8% and 1% YC groups exhibited a significant increase in Ruminococcus, Propionibacterium clostridiales, and Bifidobacterium density. The density of Bacteroides in the YC groups was significantly higher than that of the control group. On day 42, the densities of Bacteroides, Sphingomonas and Bifidobacterium were higher in the 0.8% YC group, whereas a significant decrease was observed in the number of Enterobacteriaceae. These results serve as evidence that dietary supplementation with 0.8% YC not only moderately optimized the feed efficiency and the apparent digestibility of ether extract, calcium and phosphorus, but also positively influenced the caecal bacterial density and diversity in broiler chickens.