After the legislative ban on the utilization of antibiotics in animal feed, phytochemical substances gained increasing attention as alternatives to antibiotics because of their bioactivities and safety for animals. The present study aimed to investigate the influence of dietary rosemary extract (RE) on growth performance, meat quality, serum biochemistry, antioxidant and immune capacities, and gut microbiota composition of broilers. By exploring connections among RE, physiological characteristics of broilers, and key microbiota, we sought to provide evidence for the utilization of RE in poultry feed. A total of 280 1-d-old female AA broilers were randomly separated into five groups, and were fed a basal diet supplemented with 0, 250, 500, 750, and 1,000 mg/kg of RE, respectively. Results showed that with regard to growth performance, both 500 and 750 mg/kg RE reduced the broiler feed-to-gain ratio from 1 to 21 d (P = 0.018). Regarding meat quality, all compositions of dietary RE reduced cooking loss of breast muscle (P < 0.01), and 500 and 1,000 mg/kg RE reduced the cooking loss of thigh muscle (P = 0.045). Regarding serum biochemical indexes, 500 mg/kg RE reduced ALB, TCHO, HDL-C, and LDL-C, and 750 mg/kg RE reduced GLU, TP, ALB, UA, TG, TCHO, HDL-C, and LDL-C (P < 0.01). Regarding antioxidant and immune capacities, 250, 500, 750, and 1,000 mg/kg RE increased T-AOC, GSH-Px, SOD, CAT, IL-2, IgA, IgG, and IgM levels (P < 0.01), and decreased serum MDA level (P < 0.01). RE at 750 mg/kg showed similar effects on growth performance, meat quality, and antioxidative and immune capacities, but a better influence on serum biochemical indexes of broilers compared with 500 mg/kg. Further analysis was conducted to investigate the effect of 750 mg/kg dietary RE on the gut microbial composition of broilers, and the results showed that 750 mg/kg RE reduced the relative abundance of g_Lachnoclostridium, g_Escherichia_Shigella, and g_Marvinbryantia (P <0.05, LDA score >2), which were negatively correlated to antioxidative and immune-associated parameters (P < 0.05). In conclusion, 750 mg/kg dietary RE was shown to have certain beneficial effects on growth performance and meat quality, and hypolipidemic and hypoglycemic effects on broilers. Furthermore, dietary RE improved antioxidant and immune capacities, which was partially attributed to the reduced abundance of certain pathogenic bacteria in broilers.