Tehran University of Medical Sciences Iranian Journal of Microbiology 2008-3289 18 1 2026 02 15 165 171 Suprina Sharma Central Department of Microbiology, Tribhuvan University, Kirtipur, Nepal Dev Joshi Central Department of Microbiology, Tribhuvan University, Kirtipur, Nepal Pramod Poudel Central Department of Biotechnology, Tribhuvan University, Kirtipur, Nepal Supriya Sharma Central Department of Microbiology, Tribhuvan University, Kirtipur, Nepal Tika Karki Department of Biotechnology, Kathmandu University, Kavre, Nepal Reshma Tuladhar Central Department of Microbiology, Tribhuvan University, Kirtipur, Nepal 2025 08 01 2025 12 08 Background and Objectives: Maize and peanuts, staple foods in Nepal, are highly susceptible to fungal contamination, particularly with aflatoxin-producing Aspergillus flavus. This study aimed to detect aflatoxin-producing A. flavus in maize and peanut samples collected from different regions of Nepal. Materials and Methods: A total of 80 maize and 20 peanut samples were collected randomly from markets and households. Disinfected samples were inoculated on Potato Dextrose Agar and incubated at 28°C for 48 hours. A. flavus was identified by conventional cultural methods. Aflatoxin production was screened on Aspergillus Differential Medium (ADM) and confirmed through Thin Layer Chromatography (TLC) and High Performance Liquid Chromatography (HPLC). Results: Fungal contamination was detected in 67.0% of samples, with a significantly higher rate in household-stored samples (p = 0.00039). A. flavus was the predominant species, particularly in maize (72.5%). Of 50 A. flavus isolates, 15 (30%) were aflatoxin positive on ADM, with 11 (73.3%) being confirmed by TLC. HPLC revealed AFB1 as the most prevalent in both maize and peanuts, while AFB2 was restricted to maize. Conclusion: Maize and peanuts are highly susceptible to contamination with aflatoxin producing A. flavus in Nepal, particularly in household-stored samples, emphasizing significant food safety concerns. https://ijm.tums.ac.ir/index.php/ijm/article/view/5680 https://ijm.tums.ac.ir/index.php/ijm/article/download/5680/1874