Background: Food contamination with aflatoxin is one of the most important concerns of health professionals. One of the best ways to reduce aflatoxin content in food is to use probiotics. Therefore, this study was performed to isolate Bifidobacterium from the chick intestine and its probiotic activities and also its application with Polyvinylpyrrolidone (PVP) to reduce aflatoxin B1 (AFB1) toxin in the medium were investigated.Results: Samples were isolated from the chicken intestine. Biochemical and molecular evaluations indicated isolation of Bifidobacterium bifidum strain from chicken intestine. To measure probiotic activities, pH, bile, and salt tolerance tests were used. The selected B. bifidum had good probiotic activity. Then, the antimicrobial activity of isolate against gastrointestinal pathogens and the antibiotic susceptibility test were done. The selected strain showed antimicrobial activities on S. enterica, E. coli, and P. vulgaricus and was found to be resistant against Amikacin, Ampicillin, Erythromycin, and Ceftazidine antibiotics. Then, the effect of selected isolate and PVP on reducing AFB1 in the medium was studied using ELISA and HPLC. The selected strain showed the ability to reduce the concentration of AFB1 in the medium (50% reduction) and when used in combination with PVP showed the synergistic effects in reducing the concentration of AFB1 from the medium (up to 90%).Conclusion: In conclusion, it was found that selected B. bifidum strain together with PVP could have synergistic effects in reducing AFB1 toxin in medium up to 90%.