This study was carried out to determine the efficacy of different applications of a biopesticide for reduction of aflatoxin contamination in peanut. The biopesticide, afla-guard, delivers a nontoxigenic Aspergillus flavus to the field where it competes with naturally occurring toxigenic fungus. Biocontrol treatments included: (ı) soil application during sowing, (ıı) multiple application during sowing and 40 days after planting, (ııı) foliar application at 60 days after planting (ıv) control (untreated plots). Biopesiticide was applied to peanut plots in 2015 and 2016 in Randomized Complete Block Design with four replications. Peanuts were collected from control and treated plots at harvest-drying-pre-storage periods and analysed for aflatoxins. Aflatoxin concentrations were generally quite low in 2015, also the aflatoxin concentration in treated samples (from 0.04 to 0.71 µg/kg) was reduced by 97.38 to 99.82% compared with controls (from 21.84 to 27.12 µg/kg). In 2016, reductions were also noted for all biocontrol treatments (from 89.07 to 92.39%) compared with controls. In conjunction with the reductions in aflatoxin contamination, biocontrol treatments produced significant reductions with biopesticide in peanut. Therefore, it can be said that a biological control method is a promising approach for controlling aflatoxin.