TLRs are important innate immune receptors. It has been found that vanadium can affect the immune function in broilers. However, vanadium action in the regulation of TLRs is unknown. A total of 420 one-day-old avian broilers were divided into six groups, and fed on a corn-soybean basal diet as control diet (vanadium 0.073 mg/kg) or the same diet supplemented with vanadium at the doses of 5, 15, 30, 45 and 60 mg/kg in the form of ammonium metavanadate. TLR4 and TLR7 mRNA expression in lymphoid organs of each group were studied by real time RT-PCR form. The results indicated that the TLR4 and TLR7 mRNA expression in thymus and TLR4 mRNA expression in bursa were down-regulated in the 60 mg/kg group. The TLR7 mRNA expression in bursa was down-regulated in 30 mg/kg, 45 mg/kg and 60 mg/kg groups, and up-regulated in the 5 mg/kg group. Also, the TLR4 and TLR7 mRNA expression in spleen was down-regulated in the 45 mg/kg and 60 mg/kg groups, and up-regulated in the 5 mg/kg group. The abovementioned results showed that dietary vanadium in excess of 30 mg/kg could down-regulate the TLR4 and TLR7 mRNA expression in lymphoid organs, which finally impaired innate immunity in broilers.