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Immunosuppressed individuals, including those undergoing cancer treatment, are more vulnerable to fungal infections, such as oral candidiasis, impacting their quality of life. Given the limitations of current therapies, the discovery of new antifungal agents, including those of natural origin, is crucial for the proper managing of these infections. We investigated the phytochemical profile and antifungal activity of both the essential oil and crude ethanolic extract (CEE) obtained from Eugenia luschnathiana against reference strains and clinical isolates of Candida from oncology patients. Toxicological characterization was also conducted. Gas chromatography coupled to mass spectrometry (GC-MS) and 1H Nuclear Magnetic Resonance (NMR) were used for phytochemical analysis. Antifungal evaluation was conducted to determine the Minimum Inhibitory Concentration (MIC) and Minimum Fungicidal Concentration (MFC); evaluation of potential mechanisms of action; activity on a fungal biofilm; evaluation of the cytotoxic effect on human keratinocytes of the HaCat lineage by the MTT method; determination of lethality for Artemia salina larvae. GC-MS identified a predominance of sesquiterpenes in the essential oil, notably (E)-Caryophyllene. The 1H NMR spectrum identified aliphatic, osidic, and aromatic compounds in the crude ethanolic extract. The essential oil showed no antifungal activity. However, the CEE exhibited fungicidal activity, with MIC and MFC ranging from 1.95 µg/mL to 3.90 µg/mL. The antifungal effect was affected by sorbitol, indicating a possible mechanism targeting fungal cell wall structures. At low concentration (19.5 µg/mL), the CEE inhibited 62,78% of C. albicans biofilm. The CEE demonstrated a promising toxicity profile, with an LC50 of 142.4 µg/mL against Artemia salina. In conclusion, the CEE from Eugenia luschnathiana exhibited potent antifungal activity, likely through cell wall disruption, biofilm inhibition, and a favorable toxicity profile for further exploration.