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.

Photodynamic inactivation of Salmonella enterica and Listeria monocytogenes inoculated onto stainless steel or polyurethane surfaces.

The photodynamic inactivation (PDI) uses molecules (photosensitizers) that absorb visible light (385-450 nm) energy, transfer it to adjacent molecular oxygen and thereby generating the biocidal singlet oxygen and other reactive oxygen species in situ. Efficacy of PDI was tested against Listeria monocytogenes and Salmonella enterica in three ways. Firstly, by adding the photosensitizer to bacterial suspensions. Secondly, bacteria were placed on inanimate surfaces and then sprayed with a photosensitizer suspension. Thirdly, bacteria were placed on coated inanimate surfaces, where the photosensitizer was permanently fixed in this coating (antimicrobial coating, AMC). Experiments were performed without and with soiling (albumin, sheep erythrocytes). In suspension, PDI reduced the number of viable Listeria monocytogenes and Salmonella enterica by more than 6 Log CFU/mL within seconds of light exposure. Photosensitizer spray suspension reduced the bacterial burden on surfaces with up to about 6 Log CFU/mL (5 s light exposure). PDI, even in the presence of high soiling, achieved a reduction of up to 5.1 ± 1.2 Log CFU/mL. The AMC showed a bacterial reduction that decreased from 5.1 to 0.7 Log CFU/mL with increasing soiling. Depending on the soiling and the respective bacteria, the spray suspension or AMC achieved a bacterial reduction on the running conveyor belt demonstrator ranging from 2.9 to 5.3 or 0.5 to 4.5 Log CFU/mL, respectively. PDI used visible light, phenalene-1-one and curcumin photosensitizers, and oxygen from ambient air to reduce the bioburden on typical surfaces in food processing. The AMC acts slower than the spray suspension but enables a permanent, self-sanitizing effect.

Disinfection of an ambulance using a compact atmospheric plasma device.

AIMS: The worldwide spread of the coronavirus SARS-CoV-2 has highlighted the need for fast and simple disinfection processes, amongst others for ambulance cars on site. To overcome current drawbacks regarding room disinfection, the use of cold atmospheric plasma in remote operation represents a promising alternative for the disinfection of larger volumes. In this study, a compact plasma system was evaluated regarding its disinfection efficiency inside an ambulance car. METHODS AND RESULTS: The developed plasma device is based on a dielectric barrier discharge (DBD) and operates with ambient air as process gas. The humidified afterglow from the plasma nozzle was introduced into an ambulance car with a volume of approximately 10 m3 while Bacillus atrophaeus endospores, Staphylococcus aureus or Phi 6 bacteriophages dried on different surfaces (PET-films, glass slides or aluminum foil) were exposed to the reactive gas inside the ambulance vehicle at eight different positions. Reductions of spores by more than 4 orders of magnitude were found on all surfaces and positions within 2 h. Due to their higher susceptibility, Phi 6 bacteriophages and S. aureus counts were reduced by at least 4 orders of magnitude within 30 min on all surfaces. CONCLUSION: The results show that different microorganisms dried on variable surfaces can be inactivated by several orders of magnitude inside an ambulance by plasma gas from of a compact DBD plasma nozzle. SIGNIFICANCE AND IMPACT OF THE STUDY: Plasma gas generated on site by a DBD plasma nozzle proved to be highly efficient for the disinfection of the interior of an ambulance car. Compact plasma systems could be a viable alternative for the disinfection of vehicles or rooms.

Litsea cubeba fruit essential oil and its major constituent citral as volatile agents in an antimicrobial packaging material.

Food packaging films were coated with polyvinyl acetate (PVA) containing different concentrations of citral or Litsea (L.) cubeba essential oil (EO). Antimicrobial contact trials in style of ISO22916 were performed. Citral coatings achieved bactericidal effects against Escherichia coli (2.1 log) and Staphylococcus aureus (4.3 log) at concentrations of 20%DM. L. cubeba inactivated more than 4 log cycles of both bacteria at a concentration of 20%DM. To determine the antimicrobial activity across the gas phase, a unique method for volatile agents was developed, adapting ISO22196. GC/MS measurements were performed to supplement microbiological tests in a model packaging system with a defined 220 ml headspace (HS). HS-equilibrium concentrations of 1.8 µg/mlAir were found for 20%DM 'citral-coatings, resulting in antimicrobial effects of 3.8 log against of E. coli. Saccharomyces cerevisiae (4.74 log) and Aspergillus niger (4.29 log) were more effectively inactivated by 3%DM and 5%DM coatings. In an application trial with strawberries, simulating a headspace packaging, growth inhibitory effects on the yeast and mold microbiota were found for the 20%DM coatings.

Screening essential oils for their antimicrobial activities against the foodborne pathogenic bacteria Escherichia coli and Staphylococcus aureus.

The application of essential oils as antimicrobials is a current subject of research and a promising approach in terms of natural food preservation. Due to the diversity of EO producing plant genera and the inconsistent use of susceptibility testing methods, information on the antibacterial potency of many EO varieties is fragmentary. This study was performed to assess the minimal inhibitory concentrations (MIC) of 179 EO samples from 86 plant varieties, using a single method approach, excluding emulsifying agents. MICs were acquired in a broth microdilution assay, using a dispersion based approach to incorporate EOs in a concentration range of 6400 to 50 µg/ml. Staphylococcus aureus and Escherichia coli were used as model bacteria. At concentrations below 400 µg/ml S. aureus was inhibited by 30, E. coli by 12 EO varieties. Azadirachta indica (50 µg/ml vs. S. aureus) and Litsea cubeba (50 µg/ml vs. S. aureus, 200 µg/ml vs. E. coli) essential oils were identified as promising new antimicrobial EO candidates with significant antimicrobial activity against the two foodborne pathogenic bacteria.