The composition and function of the intestinal microbiota play important roles in digestion and degradation of herbal medicines (HMs). However, few studies have examined the relationship between the fecal microbiota and HMs. In this study the effect of unfermented Astragalus (UA) and fermented Astragalus (FA) on growth performance, serum biochemical parameters, and fecal microbiota was evaluated in broiler chickens. In total, 180 one-day-old broiler chickens (Avian breeds) were randomly assigned to a control (C) group fed a basal diet, an unfermented (U) group fed a basal diet containing 0.5% UA, or a fermented (F) group fed a basal diet containing 0.5% FA, for 42 days. The F/G ratio was lower in F and U groups than in C group from 22 to 42 days (P < 0.05). Glutathione superoxide dismutase, antioxidant capacity, and total superoxide dismutase were higher, whereas malondialdehyde was lower in F group than in C and U groups from 1 to 21 days and from 22 to 42 days (P < 0.05). Fecal microbiota were profiled on an Illumina MiSeq platform following PCR amplification of the V4 region of the 16S rRNA gene. At the genus level Lactobacillus was the most abundant genus on day 7 in F group. Importantly, a potentially pathogenic genus, Enterococcus, was less abundant in the U and F groups than in the C group on day 35 (P < 0.05). These results indicate that dietary supplementation with 0.5% FA has beneficial effects on growth performance, serum biochemical parameters and fecal microbiota of broiler chickens.