The use of nano-emulsions has great advantages over conventional macroemulsions since the small droplet size allows to expand the options of applications besides presenting a greater surface area. This chapter focuses on the formulation of nano-emulsions of citrus essential oils in water, stabilized with a natural gum (mesquite gum), using a high pressure microfluidic homogenizer to obtain appropriate physicochemical characteristics and kinetic stability. When establishing the general conditions of the methods for obtaining nano-emulsions by high pressure homogenization, several formulations presented stability and size corresponding to nano-emulsions, and these were monitored during 4 months in order to study their stability as a function of time. Taking into account the results of size and stability, the best nano-emulsion obtained had a composition of Persian lemon oil (9.86%), mesquite gum (4.93%) Tween 80 (4.89%), Span 20 (1.45%), and deionized water (78.86%) with an average droplet size of 40 nm. In addition, the antibacterial activity studies also showed that this formulation had the best performance against common bacteria such as Staphylococcus aureus and Escherichia coli. The analysis of the minimum inhibitory concentration (MIC) shows that it is possible to prevent the growth of these particular bacteria using 6.25% of the best nano-emulsion formulations.