The present work was carried out to study the ability of five probiotics on the in vitro degradation of Aflatoxins B1 (AFB1). The best results of in vitro were tested on the detoxification of AFB1 in rabbits. A total of 40 growing New Zealand White (NZW) male rabbits were assigned to five experimental groups. Animals were fed the following diets: basal diet (control), basal diet contaminated with 300 ppb AFB1, basal diet contaminated with 300 ppb AFB1. + probiotic 3 (0.5 g/kg diet), basal diet contaminated with 300 ppb AFB1 + ajowan (0.5 g/kg diet), and basal diet contaminated with 300 ppb AFB1 + probiotic 3 (0.5 g/kg diet) + ajowan (0.5 g/kg diet). Live body weight significantly (P < 0.05) decreased in rabbits fed AFB1 contaminated diet compared to the control rabbits. All additives improved (P < 0.05) the live body weight. The best improvement occurred with probiotics + ajowan. The addition of probiotics increased (P < 0.05) daily body weight gain in all weeks except the first week. Adding ajowan or ajowan + probiotic led to a significant (P < 0.05) increase in live body weight gain and feed intake compared to rabbits fed AFB1 alone. The digestion coefficients of dry matter (DM), organic matter (OM), crude fiber (CF), ether extract (EE), nitrogen-free extract (NFE), and digestible crude protein (DCP) significantly (P < 0.05) decreased in rabbits fed AFB1 contaminated diet. All additives improved (P < 0.05) the digestibility coefficients of DM, OM, EE, CF, NFE, and total digested nutrients (TDN)%. The best improvement occurred with probiotics + ajowan. Concentrations of serum total protein, albumin and globulin significantly (P < 0.05) decreased in rabbits fed AFB1 contaminated diet compared with the control rabbits. In conclusion, the addition of probiotic 3 (AVI-5-BAC) + ajowan could be recommended to eliminate the toxicity of AFB1 and improve growth performance criteria in rabbits.