Stone pine (Pinus pinea L.) and maritime pine (Pinus pinaster Aiton) are classifed into the Pinaceae family. The selected species from the Cupressaceae family is maritime juniper (Juniperus oxycedrus spp. macrocarpa Sibth. & Sm.). These evergreen species' habitat is in the Mediterranean and Sub-Mediterranean climate. The isolation of essential oils was carried out by the water distillation method in the Clevenger apparatus from the dried needles of the selected species. Subsequently, the essential oils were analyzed directly by coupled system gas chromatography-mass spectrometry (GC-MS). Monoterpene α-pinene was the most abundant compound in the essential oils of maritime pine and maritime juniper (28,52 % and 64,85 %, respectively), while in stone pine essential oil, monoterpene limonene was present in highest percentage (42,57 %). The antioxidant activity determined by the FRAP and DPPH methods showed that the isolated essential oils had low antioxidant activity. Furthermore, the ability to inhibit acetylcholinesterase and butyrylcholinesterase enzymes was determined by the Ellman's method. At a maximum essential oil concentration of 22.72 µg mL -1 tested, maritime pine and maritime juniper oils showed the best results for inhibiting both enzymes. The acetylcholinesterase enzyme was inhibitited with 94.08% and 78.03%; while butyrylcholinesterase was inhibited with 35.10% and 35.90%, respectively. Antimicrobial activity was also determined by the microdilution method. Essential oils did not show strong antibacterial activity, but have shown good antifungal activity. The essential oil of the maritime juniper stands out, with an MIC value below 100 µg ml -1 against S. pyogenes, E. faecalis, L. monocytogenes and S. pneumoniae, and both types of fungi tested.