Optimization of a natural antimicrobial formulation against potential meat spoilage bacteria and food-borne pathogens: Mixture design methodology and predictive modeling.

This study is about the combined antimicrobial effect of essential oils (EOs), namely Mediterranean (MN) EO, German thyme (GT) EO, Cinnamon (CN) EO, Indian (IN) EO, Asian (AN) EO, and citrus extract (CE) against spoilage bacteria (Lactobacillus sakei, Lactobacillus curvatus, Leuconostoc mesenteroides, Carnobacterium divergens, Brochothrix thermosphacta, and Pseudomonas aeruginosa) and selected pathogenic bacteria (E. coli O157:H7, Salmonella Typhimurium and Listeria monocytogenes). Firstly, each EO and CE were screened for antibacterial activity by microdilution assay, and the most efficient antimicrobial extracts were selected based on the lowest MIC values to perform the combination assays. Afterward, a simplex-centroid mixture design was used to develop optimal antimicrobial mixtures capable of protecting meat from spoilage and pathogenic bacteria. The optimization tool allowed us to postulate models and validate them statistically as well as to create a prediction profile of the experiment. Thus, the optimal mixtures named active formulation 1 (AF1) containing MN EO/GT EO/VC EO/CE with a ratio of 1:2:2:1 and active formulation 2 (AF2) containing IN EO/AN EO/CE/VC EO with a ratio of 2:2:1:2, were developed based on the demonstration of their synergistic effect against tested bacteria. The obtained formulations at organoleptically acceptable concentrations could be applied in the preservation of meat and meat products.

Advantages of nanotechnology developments in active food packaging.

Active food packaging (AP) is an innovative technology, which provides a new system for food preservation in the food industry. The growing need towards consumer demands and market trends inspires researchers and food industries to employ AP. The advent of nanotechnology has had a significant impact on AP applications due to its distinctive physicochemical and biological properties. The incorporation of nano-sized materials into the polymer matrix has greatly improved the thermal, mechanical, gas barrier properties and compostability of biopolymers, thereby overcoming the shortcomings of bio-based film packaging. One of the great features of nano-sized materials is that they can act as release-controlling devices for active compounds in AP. The current review summarized the impact of nano-sized materials in the physical properties of the bio-based packaging films and their potential utilization and applications in AP. The safety concentrations of nano-sized materials and their possible migration from food packaging into food were also reviewed to gain a perspective view on their utilization in AP.

Silver nanoparticles-essential oils combined treatments to enhance the antibacterial and antifungal properties against foodborne pathogens and spoilage microorganisms.

Plant-derived essential oils (EOs) and commercial silver nanoparticles (AgNPs) were tested to evaluate their antibacterial and antifungal efficiency against two pathogenic bacteria (Escherichia coli O157:H7 and Salmonella Typhimurium) and three spoilage fungi (Aspergillus niger, Penicillium chrysogenum, and Mucor circinelloides). A broth microdilution assay was used to determine the minimal inhibitory concentration (MIC) of EOs and AgNPs. In the MIC assay, the cinnamon EO, Mediterranean formulation, citrus EO and spherical-shaped silver nanoparticles (AgNPs) (AGC 1, AGC 0.5, AGPP and AGPPH) showed moderate to high antibacterial and antifungal properties, with MIC ranging from 7.8 to 62.5 ppm for AgNPs and 312.5-1250 ppm for EOs against the tested bacteria and fungi. The possible interaction between the EOs and the AgNPs was determined using a checkerboard method by evaluating fractional inhibitory concentration (FIC) values. The combination of two or more EOs and AgNPs (Active combination 1: AGPPH+cinnamon EO, Active combination 2: AGC 0.5+Mediterranean formulation+citrus EO, Active combination 3: AGPP+cinnamon EO+Asian formulation+lavang EO) showed synergistic effects (FIC <1.0) against all tested bacteria and fungi. A modified Gompertz model was used to evaluate growth parameters including maximum colony diameter (A), maximum growth rate (Vm), and lag phase (λ), under the three active combinations suggested by the checkerboard method using a vapor assay. The three active combinations 1, 2 and 3 reduced the growth rate and maximum colony diameter of E. coli, S. Typhimurium, A. niger, P. chrysogenum, and M. circinelloides, and extended their lag phase from 1 to 5 days. In in situ tests with inoculated rice, the three active combinations showed a significant reduction of all tested bacteria and fungi at 27 °C for 28 days.

Mixture design methodology and predictive modeling for developing active formulations using essential oils and citrus extract against foodborne pathogens and spoilage microorganisms in rice.

The antibacterial and antifungal effects of six plant-derived essential oils (EOs) and two types of citrus extracts (CEs) were studied against two pathogenic bacteria (Salmonella Typhimurium and Escherichia coli O157:H7) and three fungi (Aspergillus niger, Penicillium chrysogenum, and Mucor circinelloides). A broth microdilution assay and checkerboard method were used to measure the minimal inhibitory concentration (MIC) of each extract and the possible interactions between them. The MIC assay showed that cinnamon EO, Mediterranean EO, Southern formulation, citrus EO, organic citrus extract (OCE), and natural citrus extract (NCE) had the highest antimicrobial and antifungal activity. The checkerboard method showed that the Mediterranean EO+OCE combination acted in synergy against all tested pathogens. A centroid mixture design was used to develop active formulations by predicting optimal concentrations of EO/CEs for increased antibacterial/antifungal activity. A mixture of four formulations (625 ppm OCE, 313 ppm Mediterranean EO, 625 ppm citrus EO, and 313 ppm cinnamon EO) named as active formulation 1, and the mixture from five formulations (625 ppm NCE, 625 ppm Asian formulation, 313 ppm Southern formulation, 625 ppm cinnamon EO, and 313 ppm savory thyme EO) named as active formulation 2, were formulated and tested because of their high microbicidal effectiveness. In situ tests with rice showed a significant reduction (P ≤ 0.05) of all tested pathogenic bacteria and fungi from the vapor of active formulations 1 and 2 after 28 days of storage. PRACTICAL APPLICATION: Active formulations (essential oils and citrus extracts) developed in the study are highly effective against foodborne pathogens. Active formulations in this study could be used as natural preservatives in the food industry for controlling foodborne diseases and spoilage organisms in stored foods.