Mechanism of Bacterial Inactivation by (+)-Limonene and Its Potential Use in Food Preservation Combined Processes

This work explores the bactericidal effect of (+)-limonene, the major constituent of citrus fruits' essential oils, against E. coli. The degree of E. coli BJ4 inactivation achieved by (+)-limonene was influenced by the pH of the treatment medium, being more bactericidal at pH 4.0 than at pH 7.0. Deletion of rpoS and exposure to a sub-lethal heat or an acid shock did not modify E. coli BJ4 resistance to (+)-limonene. However, exposure to a sub-lethal cold shock decreased its resistance to (+)-limonene. Although no sub-lethal injury was detected in the cell envelopes after exposure to (+)-limonene by the selective-plating technique, the uptake of propidium iodide by inactivated E. coli BJ4 cells pointed out these structures as important targets in the mechanism of action. Attenuated Total Reflectance Infrared Microspectroscopy (ATR-IRMS) allowed identification of altered E. coli BJ4 structures after (+)-limonene treatments as a function of the treatment pH: ß-sheet proteins at pH 4.0 and phosphodiester bonds at pH 7.0. The increased sensitivity to (+)-limonene observed at pH 4.0 in an E. coli MC4100 lptD4213 mutant with an increased outer membrane permeability along with the identification of altered ß-sheet proteins by ATR-IRMS indicated the importance of this structure in the mechanism of action of (+)-limonene. The study of mechanism of inactivation by (+)-limonene led to the design of a synergistic combined process with heat for the inactivation of the pathogen E. coli O157:H7 in fruit juices. These results show the potential of (+)-limonene in food preservation, either acting alone or in combination with lethal heat treatments

Rapid discrimination and classification of edible insect powders using ATR-FTIR spectroscopy combined with multivariate analysis

Insects are being proposed as an alternative way to ensure world''s food and feed security. Methods to determine edible insect powder''s origin and species will be needed for quality control purposes. Infrared spectroscopy has been extensively used in rapid chemical fingerprinting of food products. The present research explores a new approach to discriminate and classify commercial edible insect powders using attenuated total reflectance mid-infrared spectroscopy combined with multivariate analysis. Infrared spectra of seven commercial edible insect powders from different species (Tenebrio molitor, Alphitobius diaperinus, Gryllodes sigillatus, Acheta domesticus and Locusta migratoria) and origins (the Netherlands and New Zealand) were collected to build up soft independent modelling of class analogy (SIMCA) models. SIMCA models clearly discriminated insects by their species and origin linking their differences to lipids and chitin. SIMCA models performance was tested using five spectra of each class not used to build up the training set. 100% correct predictions were obtained for all the samples analysed with the exception of one sample of Alphitobius diaperinus. Infrared spectroscopy coupled to multivariate analysis provided a powerful method for the assurance of insect powder''s authenticity