Investigations were carried out to assess the efficiency of 3 essential oils, clove, tea tree, and rosemary, as natural preservatives during the postharvest of lettuce leaves. The effect of different concentration (1 and 0.5 MIC) of plant essential oils applied in 3 forms (spray, immersion, and capsules) was studied on lettuce leaves. The evolution of different microbial populations was evaluated during refrigerated storage. The application forms of the biopreservatives were shown to be an important factor in determining the effectiveness of the essential oils. Clove and tea tree essential oils at 1 MIC and applied embedded in lactose capsules presented a significant inhibition on mesophilic, psicrotrophic, and coliforms populations, while rosemary in none of the 3 technological applications forms exerted inhibitory effect on all microbial populations evaluated. Essential oils (at 0.5 MIC) applied by spray, immersion, and embedded in lactose capsules exerted lower inhibitory effects, with respect to 1 MIC, on the different microbial populations present on lettuce leaves. At the end of the storage (7 d), lettuce samples treated with tea tree, clove, and rosemary (at 1 and 0.5 MIC) by spray were the only organoleptically acceptable. It is concluded that clove and tea tree essential oils can control different microbial population present in lettuce. Practical Application: The exploration of naturally occurring antimicrobials in food preservation receives increasing attention due to consumer awareness of natural food products. Biopreservatives are useful in extending the shelf life of foods, reducing or eliminating pathogenic bacteria and increasing overall quality of food products. The effectiveness of essential oil application in foods is the result of factor associations such as applications forms, concentration applied, the way of action, storage temperatures. The application methods (spray, immersion, and embedded in lactose capsules) and the concentration of essential oils have been shown to be important factors in determining the effectiveness of these biopreservatives. The oil concentrations required to produce a certain level of inhibition in actual foods could be questionable due to the organoleptic impact. However, these novel natural preservatives in combination with other factors in obstacle technologies are an alternative to control the pathogen growth minimizing undesirable changes in organoleptic characteristics.