Essential oils have great potential in the field of the food industry as they can effectively prevent the presence of several bacterial and fungal pathogens. Essential oils are complex volatile compounds, synthesized naturally in different plant parts during the process of the secondary metabolism. The main goal of this work is to perform a qualitative evaluation of the antibacterial properties of 24 chemotyped essential oils against the growth of Bacillus subtillis. These Gram-positive bacteria are responsible for “rope” disease in bread preservation processes. The study was carried out using the method of disk-diffusion in agar. Biological activity was observed in five essential oils: Cymbopogon martinii var. motia, Thymus vulgaris QT Linanol, Thymus satureioides, Mentha piperita and Eugenia caryophyllus. The first three have in common the presence of some mono terpenic derivatives—Geraniol, Linalool and Carvacrol, respectively—with strong antimicrobial effects. The Cymbopogon martinii essential oil is one of the botanicals with the highest geraniol content (up to 80.53%) and showed more activity antimicrobial than the others. A contributing role of this knowledge could be the design of Cymbopogon martinii essential oil formula, which can be used in bakery industry as a preservative, such as nano-encapsulation for bakery doughs, active packaging of baked products or surface disinfectants.