Many types of Bacillus thuringiensis (Bt)-crops are being grown worldwide, triggering concerns about their potential impact on humans and livestock. To ensure better yield and food safety in China, an attempt has been made to develop Bt-rice targeting a broad range of insects. We aimed to investigate whether feeding genetically modified rice expressing the Bt chimeric Cry1Ac/Cry1Ab protein has any effects on the intestinal microbiota of broilers. Broilers were fed either Bt-rice or its unmodified isogenic parent line for 42 days, and total DNA was isolated from cecum contents for high-throughput sequencing of the 16S rRNA gene. In total, 1,241,005 reads, assigned to 12 phyla, 31 families, and 48 genera were generated. No significant differences were observed in the relative abundance of organisms identified among the major phyla, families, and genera, except for two less abundant families, Thermoanaerobacteraceae and Peptostreptococcaceae, and two less abundant genera, Anaerotruncus and Gelria. The results were in agreement with those from culture-based analysis and Biolog EcoPlates. These results illustrate that feeding Bt-rice has no adverse effects on the broiler intestinal microbiota and provide sufficient support for the food safety of Bt-rice.