The aims of this work are to investigate the chemical composition, the antibacterial activity of the essential oil and the chromosome numbers of two populations of Hertia cheirifolia. The samples were collected in the flowering stage, in eastern Algeria locality. The aerial parts of H. cheirifolia were submitted to a hydro-distillation. GC and GC / MS analysed the chemical compositions of the obtained essential oils. The antibacterial activity of essential oils was evaluated using the disks diffusion method against ten bacterial strains. For karyotypic analysis, the squashing method is used. Fifty-eight compounds representing 98.93% of the total oil were identified in H. cheirifolia. The chemical composition is dominated by the presence of major products, α-pinene (48.49 - 53.85%) and Germacrene-D (2.64 - 12.66%). Two distinct chemical breeds were identified, the α-pinene-spathulenol of Batna population, and the α-pinene-germacrene-D of Setif population. The essential oil of H. cheirifolia has a moderate activity against bacteria tested. In contrast, the strains E. coli ATCC 25922, P. syringae ATCC 53543 and E. fecalis ATCC 49452 are resistant to H. cheirifolia essential oils. The observations of root cells meristematic at metaphase of H. cheirifolia gave a diploid chromosome number 2n = 2x = 20, with a basic chromosome number (x= 10).