© 2022 Elsevier Inc.Aims: Aflatoxin B1 (AFB1) is the most toxic and common form of AF found in food and feed. Although AFB1 exposure has toxic effects on many organs, studies on the brain are limited. Moreover, to the best of our knowledge, there is no study on the effect of probiotics on AFB1-induced neurotoxicity. Therefore, we aimed to evaluate the possible effects of probiotics on AFB1-induced neurotoxicity in the brain. Main methods: Thirty-two adult male Wistar rats were divided into four groups: Vehicle (VEH), Probiotic (PRO) (2.5 × 1010 CFU/day VSL#3, orally), Aflatoxin B1 (AFB1) (25 μg/kg/week AFB1, orally), and Aflatoxin B1 + Probiotic (AFB1 + PRO) (2.5 × 1010 CFU/day VSL#3 + 25 μg/kg/week AFB1, orally). At the end of eight weeks, rats were behaviorally evaluated by the open field test, novel object recognition test, and forced swim test. Then, oxidative stress and inflammatory markers in brain tissues were analyzed. Next, brain sections were processed for Hematoxylin&Eosin staining and NeuN and GFAP immunostaining. Key findings: Probiotic supplementation tended to decrease oxidative stress and inflammatory markers compared to the AFB1 group. Besides, brain tissues had more normal histological structures in VEH, PRO, and AFB1 + PRO groups than in the AFB1 group. Moreover, in probiotic groups, GFAP immunoreactivity intensity was decreased, while NeuN-positive cell number increased in brain tissues compared to the AFB1 group. Significance: Probiotics seem to be effective at reducing the neurotoxic effects of AFB1. Thus, our study suggested that especially Bifidobacterium and Lactobacillus species can improve AFB1-induced neurotoxicity with their antioxidant and anti-inflammatory effects.