Mycotoxins are toxic secondary metabolites produced by toxigenic fungi that commonly contaminate agricultural crops. The purpose of the current study was to evaluate whether Bacillus subtilis ANSB060, Bacillus subtilis ANSB01G, and Devosia sp. ANSB714-based mycotoxin biodegradation agent (MBA) could alleviate the negative effects of naturally moldy diet containing aflatoxin (AF), zearalenone (ZEN), and deoxynivalenol (DON) on growth performance, serum immune function, and antioxidant capacity as well as tissue residues in mice. A total of 54 mice were randomly divided into three dietary treatments: basal diet (CON), multi-mycotoxins contaminated diet (MCD) containing AF, ZEN and DON and multi-mycotoxins contaminated diet plus MBA at a dose of 1.0 g kg-1 feed (MCD + MBA). Mice fed with moldy diet showed a significant decrease in body weight gain (p < 0.05), whereas the relative weight of the liver, spleen and uterus were remarkably increased (p < 0.05). Serum IgA and IgM contents were significantly decreased in MCD treatment compared with that in CON treatment (p < 0.05). In contrast, serum interleukin-1β (IL-1β), interleukin-2 (IL-2), and interleukin-6 (IL-6) concentrations were significantly promoted in mice fed with moldy diet (p < 0.05). Besides, the exposure to mycotoxins caused marked down-regulation of serum superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities in mice (p < 0.05). The addition of MBA effectively counteracted these toxic effects of moldy diet on mice. And DON residues in kidneys of mice consuming moldy diet were eliminated by the supplementation with MBA. Taken together, Bacillus subtilis ANSB060, Bacillus subtilis ANSB01G, and Devosia sp. ANSB714-based mycotoxin biodegradation agent has great potential use as a microbial additive to counteract mycotoxins contamination in food and feed.