The objective of this research was to evaluate the antimicrobial activity of carbon dioxide extracts of the unicellular biflagellated green alga Dunaliella salina against Escherichia coli, Staphylococcus aureus, Candida albicans, and Aspergillus niger. The effects of different extraction pressures ranging from 185 to 442 bar and extraction temperatures ranging from 9.8 to 45.2 degrees C on the extracts' composition and consequently on their antimicrobial activities were investigated. The extracts were analyzed by gas chromatography-mass spectrometry in order to identify the compounds responsible for the antimicrobial activity detected. Fourteen different volatile compounds and several fatty acids were identified. The highest antimicrobial activity was obtained using 314 bar and 9.8 degrees C. Under these conditions, the presence of an indolic derivative that had never been reported in D. salina was detected in the extract, together with polyunsaturated fatty acids and compounds related to carotene metabolism, such as beta-ionone and neophytadiene, with known antimicrobial activity.