We investigated the in vitro synergistic antifungal potential of combining serotonin (5-HT) and sertraline with amphotericin B and itraconazole against clinical isolates of Aspergillus spp. Synergy tests were performed using the chequerboard microdilution method. Activity was measured against Aspergillus fumigatus (n = 7), Aspergillus flavus (n = 3) and Aspergillus terreus (n = 2), and compared with that for Candida albicans and Candida parapsilosis. The fractional inhibitory concentration (FIC) indices ranged between 0.25 and 3 for the various isolates tested. 5-HT was shown to enhance the activity of amphotericin B against Aspergillus spp. Combination studies with 5-HT and itraconazole and with sertraline and itraconazole or amphothericin B showed different activities for the various strains, including synergism (FIC < 1.0), additivity (FIC = 1), and indifference (FIC between 1.0 and 2.0). 5-HT and sertraline showed antagonistic activity (FIC > 2) with amphotericin B and itraconazole against C. parapsilosis.