Determination of the efficacy of ultrasound in combination with essential oil of oregano for the decontamination of Escherichia coli on inoculated lettuce leaves

Recent outbreaks related to fresh produce have shown that there is a need to develop new efficient and effective decontamination technologies that protect consumers' safety. Ultrasound and assisted ultrasound, i.e., the combination of ultrasound with another decontamination technology has been shown to be effective against bacteria. In this study equal surfaces of free-bacteria of Romaine lettuce were inoculated with Escherichia coli O157:H7 NCTC 12900. Leaves were then treated with an ultrasound probe system (26 kHz, 90 μm, 200 W, 14 mm Ø). Same processing time (300 s) was applied with continuous and pulsed modes of: (i) 10 s on/6 s off, (ii) 5 s on/5 s off and (iii) 2 s on/8 s off. Thus, total treatment durations were 5, 8, 10 and 25 min. Moreover, concentrations of essential oil (i.e. 0.010% v/v, 0.014% v/v. 0.018% v/v, 0.022% v/v and 0.025% v/v) of oregano were tested, operating with continuous and 2 s on/8 s off pulsed mode. The structure of treated samples was also analysed by SEM and light microscopy. Continuous ultrasound treatments led to a reduction of 2.65 ± 0.23 log CFU/cm² of E. coli on the lettuce. No significant differences were found among the tested pulsed and continuous modes. In addition no significant differences were found between injured and no-injured cells among all the applied treatments on the leaves of the lettuce. Application of the different concentrations of EO alone, at the levels mentioned previously reduced the bacteria on the surface of the lettuce. Less than 2 logs of total inactivation (on the lettuce and the treated water) were achieved when different concentrations of EO were tested alone. Assisted ultrasound enhanced the levels of microbial reduction on the surface of the lettuce at concentrations higher than 0.010% (v/v) of oregano EO. Significant differences were found at 0.018%, 0.022% and 0.025% (v/v) both in continuous and pulsed US modes when compared with samples treated only with US. Same concentrations of EO combined with continuous or pulsed mode of US reduced the level of E. coli in water below the limit of detection. When continuous or pulse mode of US was combined with 0.025% (v/v) of EO of oregano the result was synergetic. Images reported neither structural differences nor damage of the leaves among treated samples and controls. Concluding, the effectiveness of the assisted ultrasound process was dependent on the processing time rather than on the different configurations (continuous or on/off) or the total treatment duration. Furthermore, EO combined with US could work synergistically on enhancing the inactivation of the tested bacteria.