Tehran University of Medical Sciences Iranian Journal of Microbiology 2008-3289 17 2 2025 04 05 303 311 Mahnour Taghavi Department of Microbiology, Faculty of Pharmacy, Islamic Azad University, Tehran, Iran Maryam Ahmadi Department of Microbiology, Faculty of Pharmacy, Islamic Azad University, Tehran, Iran Davoud Dehghan-Nayeri Department of Microbiology, Faculty of Pharmacy, Islamic Azad University, Tehran, Iran Zahra Salehi Department of Mycology, Pasteur Institute of Iran, Tehran, Iran Masoomeh Shams-Ghahfarokhi Departmentt of Mycology, Faculty of Medical Science, Tarbiat Modares University, Tehran, Iran Fatemehsadat Jamzivar Department of Mycology, Pasteur Institute of Iran, Tehran, Iran Mehdi Razzaghi-Abyaneh Department of Mycology, Pasteur Institute of Iran, Tehran, Iran 2024 10 19 2025 02 14 Background and Objectives: The increasing prevalence of fungal infections due to antifungal resistance underscores the need for novel treatment strategies. The present study aimed to investigate the inhibitory effects of soil-originated antagonistic bacteria against Aspergillus and Trichophyton species. Materials and Methods: Fifty soil samples collected from Isfahan and Khuzestan provinces by using the Zig-Zag method were cultured on glucose-yeast extract (GY) agar around fungal colonies to isolate antagonistic bacteria. Antifungal activity was assessed by measuring clear zones around the colonies of A. niger, A. fumigatus, T. rubrum, and T. mentagrophytes by co-culture linear method. Potent antagonistic bacteria were identified by 16S rRNA sequencing, and evaluated for antifungal activity using disk diffusion assays compared with amphotericin B and ketoconazole. Results: Among 50 samples, fifteen showed antifungal effects, yielding 55 bacterial strains. Four isolates with strong antifungal activity against all tested fungi were identified as Bacillus subtilis, B. licheniformis, B. axarquiensis, and Bacillus sp. These bacteria were distributed in distinct clusters phylogenitically and showed diverse antifungal activity. Conclusion: The results suggest the potential of soil-derived Bacillus species as promising antifungal agents. Further studies are recommended to identify their inhibitory metabolites, their ability as biocontrol agents against soil habitated fungi and to explore their mechanism of action and spectrum of activity. https://ijm.tums.ac.ir/index.php/ijm/article/view/5092 https://ijm.tums.ac.ir/index.php/ijm/article/download/5092/1770