The work focuses on the valorization of the plant Cotula cinerea Del., which grows spontaneously in Algeria. It is collected from two stations in the province of El Oued, to assess the chemical composition of essential oils, from two different zones whereas Beni Guecha is a reg unlike Sidi Aoun is a sandy area characterized by the presence of dunes, and to test biological activities. Oil analysis by CG/MS enabled the identification of 28 terpene components representing 96.94% ± 2.35 of the total oil. The analyzed samples differ significantly in their chemical composition. Dominated by high levels of oxygenated monoterpenes (78% and 41%), the oils are rich in 1.8-cineole, camphor, trans-thujone, cis-chrysanthenol and terpinen-4-ol. Firstly, the antibacterial potential of C. cinerea essential oils showed good inhibition against E. coli ATCC25922 and Staphylococcus aureus ATCC25923 with an inhibitory diameter ranging from (16.70–14.64 mm). In addition, the results of the antioxidant effect of these oils showed that they have moderately weaker antioxidant power than ascorbic acid. On the other hand, these EOs showed significant antifungal potential against the two fungi tested.