Ferula vesceritensis is indigenous to Algerian Sahara. According to ethnobotanical investigations, the fruit decoction is used in folk medicine to treat headaches, fever, and throat infections. In our continuing investigation of the roots of F. vesceritensis, the methylene chloride–methanol (1:1) extract of the dried roots led to the isolation of ten daucane sesquiterpenes substituted at positions 6 and 10 by angelate, p-hydroxy- or methoxybenzoic, and acetate, which were identified for the first time in this species. Evaluation of the antibacterial and antifungal activities of the crude extract using the disk diffusion method against five human pathogenic bacteria (including Gram-positive and Gram-negative) and two fungi (Alternaria alternata and Aspergillus fumigatus) showed it prevented the growth of all tested microorganisms, with average inhibition zone diameters ranging from 6.00–24.00 mm depending on concentration and microorganism type. No effect was observed against some microorganisms at the low concentration of 250 g/mL. The phytochemical findings and antimicrobial results justified and partly supported the plant's popular use as traditional remedies. Notably, the crude extract's strong activity against clinical and standard microorganisms, especially major pathogens like Proteus vulgaris and Alternaria alternata, suggests it could be a new medicinal resource for antibacterial agents.