Compared with traditional disinfectants, alternative antimicrobials have been given increasingly more attention. Alternative agents are often considered less toxic, more environmentally friendly and more natural. This group includes Thieves Household Cleaner. The key ingredients of this cleaner are non-ionic surface active agents, anionic surfactants and Thieves Essential Oil. Thieves Essential Oil is a therapeutic combination of 5 essential oils: Eugenia caryophyllus (clove), Citrus limon (lemon), Cinnamomum zeylanicum (cinnamon), Rosmarinus officinalis (rosemary) i Eucalyptus radiata (eucalyptus). This concentrated formula is suitable for almost every surface, including carpets, floors, glass and walls. The aim of this research was to examine the antibacterial effect of the universal cleaning agent Thieves on bacteria of the genera Staphylococcus aureus and Acinetobacter baumannii. Two methods were used: the method of diffusion into agar from the holes and the method of determining antimicrobial activity at a given time in which the efficacy of three differently prepared solutions was measured in a time range of 1, 3, 5, 10, 15, 20, and 30 minutes. Three different solutions were used: Thieves and vinegar, Thieves and water, and water and vinegar. All three solutions show different antimicrobial effects. Solution a) (Thieves and water) exhibits the weakest antibacterial property, while solution b) (Thieves and vinegar) exhibits the strongest antibacterial properties for all strains. In conclusion, Thieves Household Cleaner shows significant antimicrobial effects on bacteria of the genera Staphylococcus and Acinetobacter, especially the combination of Thieves and vinegar, and thus has great potential as an alternative cleaning agent that can replace coarse chemicals.