Mycotoxins like aflatoxins pose a significant threat to the health of both people and animals because of their deadly effects. This study aimed to investigate the potential of Lactobacillus strains in reducing the toxicity caused by aflatoxins in mice receiving a diet contaminated with aflatoxins. The mice were split up into various treatment groups, including a control group, an aflatoxin-treated group, and groups treated with the aflatoxin-contaminated diet along with Lactobacillus strains. Various parameters, including liver enzymes, blood parameters, malondialdehyde (MDA) levels, morphometric analysis of ileum, and gene expression, were analyzed to assess the effectiveness of the Lactobacillus strains in mitigating aflatoxins toxicity. Results showed that mice in the aflatoxin-treated group had increased MDA levels, indicating oxidative stress. Alternatively, the Lactobacillus cocktail treatment group showed a decreasing trend in MDA levels, suggesting a reduction in lipid peroxidation. The morphometric analysis of ileum tissue demonstrated that the Lactobacillus-treated group exhibited improved structural integrity compared to the aflatoxin-treated group. Additionally, gene expression analysis revealed that the Lactobacillus treatment attenuated the downregulation of SOD gene expression and mitigated the upregulation of iNOS gene expression induced by aflatoxins. These findings suggest that Lactobacillus strains have the potential to reduce aflatoxin-induced toxicity by alleviating oxidative stress, preserving intestinal tissue integrity, and modulating gene expression associated with antioxidant defense and inflammation. This study provides evidence for the beneficial effects of Lactobacillus strains in reducing aflatoxin toxicity in mice. The findings obtained may contribute to the development of preventive or therapeutic strategies against mycotoxin-induced toxicity.