This work aimed to evaluate the chemical composition of essential oils from Catha edulis Forsk collected in the Day Forest of the Republic of Djibouti. Additionally, in vivo toxicity studies, biochemical profiling, and hematological tests were conducted to determine the biological activity of the investigated essential oils. Finally, in vitro tests were performed to investigate the antibacterial activity of the essential oils. The essential oils were obtained at yields of 0.75%. Chromatographic analysis identified 39 compounds, of which cathinone (81.4%) and cathine (10.55%) were determined as the two major components, representing 91.95% of the total composition. Catha edulis essential oil had a rat LD50 of 2500 mg/kg, indicating very low toxicity. Chronic exposure studies revealed that use of the essential oil in rats resulted in persistent stimulant action at dosages of 100 and 200 mg/kg, whereas the weight gain of control rats was faster than that of the essential oil-treated rats. Hematological parameters showed a significant increase in red blood cell counts, hemoglobin, hematocrit, and platelets in treated rats, which could indicate blood hyperviscosity that may increase the risk of blood clots and cardiovascular diseases. Furthermore, the investigated essential oil exhibited antimicrobial activity against several bacterial strains.