Edible desert truffles are obligate symbiotic macro hypogenous ascomycetes fruit bodies. These are growing in depth between 5 and 10 cm. These consider a miracle of nature and an unexploited source of therapeutic compounds with antimicrobial, antioxidant, and wonderful food, especially for Bedouins. The current study investigated the activity of premature and mature ethyl acetate truffle extracts (white and Red) collected from the Western Egyptian Desert as antimicrobial and antioxidant. This study evaluated in vitro the efficacy of antimicrobial activity of organic truffles extract, and their effect on various pathogens (Gram-negative, Gram-positive bacteria, filamentous fungi, and yeast) by using agar well diffusion. TEM micrographs had been done for the most effective crude extracts. Furthermore, the activity of DPPH scavenging was studied for both mature truffles. Both truffles extracts had antibacterial activity more than antifungal activity. The selected extracts exhibited an inhibitory effect on the cell wall and protoplasm of pathogens. Terfezia sp. had antioxidant activity more than Tirmania sp. This investigation concluded that the truffle extracts could be considered a promising antibiotic and antioxidant drug in near future.