Inactivation of spoiling microorganisms in apple juice by a combination of essential oils’ constituents and physical treatments

A combination of different hurdles, such as mild heat (54 ℃ for 10 min) or pulsed electric field (25 pulses; 25 kV/cm; 3.35 kJ/cm per pulse) treatments and essential oils constituents (carvacrol, citral, and (+)-limonene), to reduce spoiling bacteria and yeasts in apple juice was evaluated. For this purpose, the heat and pulsed electric field resistances of five strains of Leuconostoc spp. and five Saccharomyces spp. strains were assayed, achieving different inactivation levels for each treatment and strain. For instance, Leuconostoc fallax 74, the most heat-resistant strain, was the second-most sensitive strain to pulsed electric field. The most resistant strains were exposed to combined processes of heat or pulsed electric field and 0.2 µl/ml essential oils constituents. The combination of heat and essential oils constituents proved to be synergistic against both microorganisms in apple juice. The most effective was the combination of mild heat and carvacrol, which caused the inactivation of 99% of L. fallax 74 and 99.99% of Saccharomyces cerevisiae CECT 1172 cells. Therefore, this study shows the great potential of carvacrol, citral, and (+)-limonene in combined treatments with mild heat to achieve a higher degree of inactivation of spoiling microorganisms in apple juice, and thus, to extend its shelf life.