Background: Aflatoxins are harmful mycotoxins that can contaminate animal and human food products. To prevent toxigenic fungi growth and aflatoxin production, researchers have long investigated plant compounds as potential inhibitory agents.Objectives: This study aimed to evaluate the in vitro effect of berberine sulfate and berberine chloride on the growth and aflatoxin production of Aspergillus flavus and Aspergillus parasiticus.Methods: The antifungal activity of berberine salts was determined according to the Clinical and Laboratory Standards Institute (CLSI) document M38-A3. The aflatoxin levels were measured using high-performance liquid chromatography (HPLC) method.Results: The berberine sulfate and berberine chloride showed inhibitory effects against both Aspergillus species, with minimum inhibitory concentration (MIC) ranging from 125 to 500 µg/mL. Berberine sulfate at 2000 μg/mL and berberine chloride at 1000 μg/mL completely inhibited the mycelial growth of A. flavus, while berberine chloride at 1000 μg/mL also completely inhibited the mycelial growth of A. parasiticus. Berberine sulfate at 2000 µg/mL reduced the mycelial growth of A. parasiticus by 96.7%. Conclusion: Berberine salts significantly decreased the total aflatoxin production of Aspergillus species at MIC/2 and MIC/4 concentrations (P˂0.05). The results suggest that berberine salts could be used as potential antifungal and anti-aflatoxigenic agents against toxigenic Aspergillus isolates.