Due to the increased resistance of bacteria to chemical drugs and the low side effects of medicinal plants, using these plants has been considered in disease treatment. Ferula asafoetida L., as one of the medicinal and native plants of southern Khorasan province (Iran), is used in traditional medicine to treat many diseases. The present study aimed to investigate the antimicrobial activity of hydroalcoholic extract of different parts of Ferula asafoetida L. in three regions of South Khorasan Province.The required chemicals were purchased from Merck and Sigma companies. The microbes were obtained from the Birjand University of Medical Sciences and the Iranian Scientific and Industrial Research Organization. The antimicrobial activity of hydroalcoholic extracts was evaluated by dilution in agar. For this purpose, the plant was collected from three areas: Sarbisheh, Ghayen, and Nehbandan. After identification, drying, and pulverizing, the extracts were prepared by soaking. Some of the extracts showed an inhibitory effect on gram-positive bacteria (Staphylococcus aureus and Bacillus subtilis), but none inhibited the growth of Gram-negative bacteria. Moreover, hydroalcoholic extracts of different parts of Ferula asafoetida L. of Sarbisheh had no inhibition effect on the growth of Candida albicans. The results revealed that the hydroalcoholic extracts of the plant could be used as a suitable substitute for chemical drugs to treat diseases. Manuscript profile