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Essential oils (EOs) play some important roles as antimicrobial and antioxidant compounds. EOs act at several cell sites and target numerous subcellular organelles such as mitochondria because of its importance in a triggering of many pathologies. However, their subcellular toxic mechanism is still under investigation. This study is carried out in order to examine the effects of Citrus aurantium L essential oilon the morphology and function of mitochondria isolated from yeast cells S. Cerevisiae. Treatment with EOs extracted from C. aurantium affects the oxygen consumption of mitochondria respiration chain by a decrease in the rate of ADP phosphorylation (inhibition of State 3). Strong Stimulation of oxygen consumption in state 4 is observed. A decrease in the respiratory control ratio (CR) obtained is due to the disturbances observed in oxidations rates of states 3 and 4. At the same time, exposition to EOs extracted from C. aurantiumdepressed the mitochondrial membrane potential (∆Ψ) by the same manner as protonophore CCCP. In parallel, EOsinduce a reduction about 80% in ATP activity. Moreover, EOs treatment generated an important oxidative stress observed with all stimulation of bioindicators stress parameters (CAT, GSH and MDA). These results suggest that mitochondrial damage observed, resulting in the uncoupling of the oxidative phosphorylation, ATP synthesis, CR and (∆Ψ) which is the fundamental factor for the oxidative phosphorylation via the mitochondrial complex V, probably due to ROS produced during oxidative stress localized on the mitochondrial respiratory chain and which could be involved in the cytotoxic activity mechanism of EOs . Finally, EOs extracted from C. aurantiumseem to exert their cytotoxic effects on the mitochondrial respiratory chain, probably by ROS generated disrupting the functioning of the mitochondrial complex V responsible for the energetic interactions, which are essential for physiological cell function.