The essential oil from Callistemon viminalis (EOC) is rich in monoterpenes, with a variety of biological properties: antibacterial, antifungal, insecticide, and antioxidant. Inclusion complexes (ICs) with cyclodextrins (CDs) is an alternative to prevent toxicity, improve the activity, and reduce the concentration to be used. Thus, the objective of this work was to prepare an IC (EOC/β-CD) and evaluate the antibacterial, antifungal and phospholipase activities, as well as the toxicity. Antimicrobial activity used the agar diffusion test and antifungal activity the disc diffusion test. Toxicity tests were carried out using Lactuca sativa L. The inhibition of phospholipase activity using the venom of Bothrops atrox as an inducer was performed. Antibacterial and antifungal tests demonstrated a decrease in the minimum inhibitory concentration (MIC) of the IC. It was most significantly observed for the bacterium Listeria monocytogenes, for which there was a decrease in the MIC from 250 µg mL-1 to 62.5 µg mL-1 after complexation, and for the fungus Aspergillus flavus, with a decrease in MIC from 125 µg mL-1 to 62.5 µg mL-1 after complexation. Toxicity tests with Lactuca sativa showed a decrease in toxicity after complexation in all parameters analyzed, with no statistical difference from the negative control. Inhibition of phospholipase activity induced by Bothrops atrox venom was more expressive in the highest proportion studied (1:10 m:m), exerting 23% inhibition. The assays demonstrated that the complexation between the EOC and β-CD is a promising alternative for use in different branches, especially in food industry, to fully exploit its application potential.