The aim of this study was to investigate the efficacy of selenium nanoparticles (SeNPs) that had been biologically synthesized by Bacillus licheniformis to counteract deoxynivalenol (DON) toxicity in laying hens. Ninety-six healthy, 20-week-old laying hens were randomly assigned to four treatment groups, each of which included 3 replicates of 8 layers, and fed four different diets: an uncontaminated basal diet (control group), a 10 mg/kg DON-contaminated basal diet (DON group), a 10 mg/kg DON-contaminated basal diet with 0.5 mg/kg SeNPs (DON+SeNPs group) and the basal diet with 0.5 mg/kg SeNPs (SeNPs group).Serum T-AOC and GPx activities were significantly decreased (P<0.05) in hens fed the DON-contaminated diet, and DON significantly decreased the egg production rate (P<0.05), significantly increased the soft-shelled or cracked egg rate (P<0.05), significantly decreased serum calcium and inhibited the immune systems of the animals according to blood routine indexes. However, SeNPs improved the levels of GPx and T-AOC, increased the egg production rate, significantly decreased the soft-shelled or cracked egg rate (P<0.05), and decreased the influence of DON on the blood routine.In addition, SeNPs significantly (P<0.05) increased serum calcium. However, no differences were observed in egg quality (egg weight, Haugh units, yolk color, eggshell strength and eggshell thickness) among the four groups. These results showed that SeNPs could provide effective anti-oxidative protection against DON toxicity in laying hens, reduced DON's influence on egg production and blood calcium