Coating with phenolic branched-chain fatty acid reduces Listeria innocua populations on apple fruit.

An antimicrobial coating was produced by mixing phenolic branched-chain fatty acid (PBC-FA) with glycerol and a carboxymethyl cellulose solution (CMC) at pH 7. The resulting PBC-FA-CMC solution formed an emulsion with an average droplet size of 77 nm. The emulsion in the coating solution was stable for at least 30 days at 20 °C. The in vitro antimicrobial activity of the film formed from the PBC-FA emulsion was tested against a mixture of 3 strains of Listeria innocua (7 log CFU/mL). Film with a concentration of 1000 µg/mL of PBC-FA effectively reduced the population of L. innocua below the limit of detection (<1.48 log CFU/mL) in vitro. The effect of the 1000 µg/mL PBC-FA-CMC coating formulation was then evaluated against L. innocua inoculated on "Gala" apples. Results showed that compared with the non-coated control, the coating reduced L. innocua populations by ~2 log CFU/fruit and ~6 log CFU/fruit on the apple when enumerated on tryptic soy agar and selective media (PALCAM), respectively, indicating that PBC-FA applied as a coating on apples resulted in the sub-lethal injury of bacterial cells. When L. innocua was inoculated onto PBC-FA-coated apples, the L. innocua population decreased by ~4 log CFU/fruit during 14 days of shelf-life at 20 °C. The PBC-FA coating lowered the moisture loss but did not affect the color, firmness, or soluble solids content of apples during the 14-day at 20 °C. Overall, this study revealed that there is a potential that PBC-FA can be used as an antimicrobial coating to inactivate Listeria and preserve the quality of apples.

Bio-based phenolic branched-chain fatty acid in wash water reduced populations of Listeria innocua on apple fruit.

Phenolic branched-chain fatty acid (PBC-FA) emulsion was produced by dissolving it in ethanol and mixing with water (pH 7). The resulting monodispersed emulsion droplets were approximately 200 nm in diameter. The stability of the emulsion was evaluated by storing it at 4 and 20 °C for 30 days. The antimicrobial activity of the PBC-FA emulsion was tested against Escherichia coli and Listeria innocua (8 log CFU/mL) by determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) using a microdilution method. The PBC-FA was effective against L. innocua with MIC and MBC of 14.1 µg/mL and caused membrane permeation as determined with SEM and Live/Dead cell assay, but was not effective against E. coli O157:H7 at the tested concentrations (5-250 µg/mL). We also evaluated PBC-FA emulsion's potential to be used as a wash against L. innocua inoculated on apples. The results showed that the 500 µg/mL PBC-FA emulsion with 5 % ethanol had equivalent antimicrobial activity (2-3 logs reductions) against L. innocua as the 20 µg/mL chlorine solution, a commonly used sanitizer. 500 µg/mL PBC-FA emulsion had better antimicrobial efficacy when organic matter (chemical oxygen demand: 9.0 g/L) was present compared to 20 µg/mL of chlorine. The effect of PBC-FA on the quality of the apples, was determined by measuring changes in color, firmness, and soluble solids content over a 14-day storage period at 20 °C. The quality of the apples was not affected by PBC-FA over the 14-day storage period, suggesting that PBC-FA emulsion can be used as a wash for apples without affecting their quality.

Green active coating from chitosan incorporated with spontaneous cinnamon oil nanoemulsion: Effects on dried shrimp quality and shelf life.

Green active film from chitosan (C) incorporated with spontaneous emulsified cinnamon oil nanoemulsion (CONE; droplet size of 79.27 nm and polydispersity index of 0.27) was developed. The obtained chitosan film containing CONE (C + CONE) had tensile elongation and light protective effect higher than C film due to the incorporation of bioactive compounds from cinnamon oil as proven by Fourier Transform Infrared Spectroscopy. The effect of C + CONE as active edible coating on the physical, chemical, and microbiological properties of dried shrimp was then investigated. The quality of samples coated with C + CONE (DS + C + CONE) was compared to those coated with C (DS + C) and without coating (DS). In this study, C + CONE could enhance astaxanthin content and reduce lipid oxidation in dried shrimp. During 6 weeks of storage, C + CONE was found to be an effective antimicrobial coating that significantly inhibited growth of bacteria, delayed lipid oxidation and retarded the production of volatile amines in dried shrimp. DS + C + CONE had lower malonaldehyde equivalents (0.52 mg/kg oil), trimethylamine (11.74 mg/100 g), total volatile base nitrogen (84.33 mg/100 g) and total viable count (4.80 Log CFU/g), but had higher astaxanthin content (12.53 ± 0.12 µg/g) than DS and DS + C. The results suggested that the developed C + CONE coating has potential to be used as active coating for preserving food quality.

Mechanism of Synergistic Photoinactivation Utilizing Curcumin and Lauric Arginate Ethyl Ester against Escherichia coli and Listeria innocua.

This study investigated the mechanism of how lauric arginate ethyl ester (LAE) improves the photoinactivation of bacteria by curcumin after diluting the 100 µmol/L stock curcumin-LAE micelle solution to the concentration used during the treatment based on the curcumin concentration. The photoinactivation of bacteria was conducted by irradiating the 1 µmol/L curcumin-LAE solution containing cocktails of Escherichia coli and Listeria innocua strains (7 log CFU/mL) for 5 min with UV-A light (λ = 365 nm). The changes in solution turbidity, curcumin stability, and bacterial morphology, viability, and recovery were observed using SEM, TEM, and live/dead cell assays. The study found that LAE enhances the photoinactivation of bacteria by increasing the permeability of cell membranes which could promote the interaction of reactive oxygen species produced by photosensitized curcumin with the cell components. The combination of curcumin and LAE was demonstrated to be more effective in inhibiting bacterial recovery at pH 3.5 for E. coli, while LAE alone was more effective at pH 7.0 for L. innocua.

Sustainable bio-based antimicrobials derived from fatty acids: Synthesis, safety, and efficacy.

Some conventional sanitizers and antibiotics used in food industry may be of concerns due to generation of toxic byproducts, impact on the environment, and the emergence of antibiotic resistance bacteria. Bio-based antimicrobials can be an alternative to conventional sanitizers since they are produced from renewable resources, and the bacterial resistance to these compounds is of less concern than those of currently used antibiotics. Among the bio-based antimicrobial compounds, those produced via either fermentation or chemical synthesis by covalently or electrovalently attaching specific moieties to the fatty acid have drawn attention in recent years. Disaccharide, arginine, vitamin B1, and phenolics are linked to fatty acids resulting in the production of sophorolipid, lauric arginate ethyl ester, thiamin dilauryl sulfate, and phenolic branched-chain fatty acid, respectively, all of which are reported to exhibit antimicrobial activity by targeting the cell membrane of the bacteria. Also, studies that applied these compounds as food preservatives by combining them with other compounds or treatments have been reviewed regarding extending the shelf life and inactivating foodborne pathogens of foods and food products. In addition, the phenolic branched-chain fatty acids, which are relatively new compounds compared to the others, are highlighted in this review.

Evaluation of the efficacy of antimicrobials against pathogens on food contact surfaces using a rapid microbial log reduction detection method.

Microbial contamination of food contact surfaces in food processing industries is a significant health hazard. Evaluating the efficacy of sanitizing agents used during food processing is essential to ensure public health and safety. This study describes an optical screening method using an oCelloScope to quantify the number of surviving bacterial cells, expressed as microbial log reduction (MLR), after antimicrobial treatment. We tested the efficacy of two sanitizing agents, sodium hypochlorite and benzalkonium chloride, against desiccated cells of three pathogens, S. Enteritidis, E. coli O157: H7, and L. monocytogenes that are of concern on food processing surfaces. Stainless steel slides were used to mimic commercial food processing surfaces. Bacterial cells were desiccated at 75% relative humidity (RH) before antimicrobial treatment on stainless steel surfaces, and survivor levels were analyzed via plate counts to calculate MLR. These were compared to MLR values generated using the oCelloScope. For analysis of MLR using the oCelloScope, cells were desiccated at 75% RH on polystyrene microtiter plates, treated with antimicrobials, and surviving cell numbers were analyzed. Our results show that MLR values of treated desiccated cells calculated using the BCA algorithm of the oCelloScope were comparable to the values generated using the traditional plate count assay for the same concentration and treatment duration of the antimicrobials against all the tested pathogens. MLR could not be calculated for a non-lytic antimicrobial (curcumin and UV-A irradiation) against E. coli O157:H7, however, modified growth curves demonstrated an antimicrobial effect of curcumin and irradiation treatment. The results indicate that this method can be used for rapid screening of MLR of lytic antimicrobial compounds. Quantification of MLR using the oCelloScope is an effective tool to rapidly identify appropriate antimicrobial treatments and can be used to study novel antimicrobial compounds in the future.

Use of Micellar Delivery Systems to Enhance Curcumin's Stability and Microbial Photoinactivation Capacity.

Microbial photoinactivation using ultraviolet (UV) or visible light can be enhanced by photosensitizers. This study assessed the efficacy of encapsulating a food-grade photosensitizer (curcumin) in surfactant micelles on its water dispersibility, chemical stability, and antimicrobial activity. Stock curcumin-surfactant solutions were prepared with Surfynol 465 (S465) or Tween 80 (T80) (5 mM sodium citrate buffer). The antimicrobial activity of curcumin-loaded surfactant solutions was determined by monitoring the inactivation of Escherichia coli O157: H7 and Listeria innocua after 5-min irradiation with UV-A light (λ = 365 nm). The solutions mixed with the bacterial suspensions contained 1 µM curcumin and each surfactant below, near, and above their critical micelle concentrations (CMCs). The addition of surfactants at any level to the curcumin solution enhanced its dispersibility, stability, and efficacy as a photosensitizer, thereby enhancing its antimicrobial activity. Gram-positive bacteria were more susceptible than Gram-negative bacteria when curcumin-loaded micelles were used against them. The photoinactivation efficacy of curcumin-surfactant solutions depended on the pH of the solution (low > high), surfactant type (S465 > T80), and the amount of surfactant present (below CMC ≥ near CMC > above CMC = unencapsulated curcumin). This result suggests that excessive partitioning of curcumin into micelles reduced its ability to interact with microbial cells. Synergistic antimicrobial activity was observed when S465 was present below or near the CMC with curcumin at pH 3.5, which could be attributed to a more effective interaction of the photosensitizer with the cell membranes as supported by the fluorescence lifetime micrographs. The use of a micelle-based delivery system facilitates adsorption and generation of reactive oxygen species in the immediate environment of the microbial cell, enhancing photoinactivation.

Impact of ripening inhibitors on molecular transport of antimicrobial components from essential oil nanoemulsions.

The objective of this study was to provide insights into the mechanisms involved in the mass transport of antimicrobial compounds from essential oil nanoemulsions to bacterial cell membranes. Origanum oil-in-water nanoemulsions were produced using spontaneous emulsification by titrating a mixture of essential oil, ripening inhibitor, and surfactant (Tween 80) into 5 mM sodium citrate buffer (pH 3.5). Stable nanoemulsions containing relatively small droplets (d < 60 nm) were produced using this low-energy method. The nature of the ripening inhibitor used in the oil phase of the nanoemulsions affected the antimicrobial activity of the nanoemulsions: corn (LCT) > medium-chain triglycerides (MCT). Differences in antimicrobial activity were attributed to the differences in the rate of transfer of hydrophobic antimicrobial constituents from the nanoemulsion to the MCT emulsion, which was used to mimic the hydrophobic region of the bacterial cell membranes. Each antimicrobial nanoemulsion was separated from the MCT emulsion by a dialysis tubing. Dialysis tubing with two different pore sizes was used, one excluding nanoemulsion droplet and micelle delivery, allowing the delivery of antimicrobial compounds only through the aqueous phase and the other by both the aqueous phase and micelles. For origanum oil nanoemulsions, the delivery of all antimicrobial agents occurred more efficiently when micelles were present.

Effect of ripening inhibitor type on formation, stability, and antimicrobial activity of thyme oil nanoemulsion.

The objective of this research was to study the impact of ripening inhibitor level and type on the formation, stability, and activity of antimicrobial thyme oil nanoemulsions formed by spontaneous emulsification. Oil-in-water antimicrobial nanoemulsions (10 wt%) were formed by titrating a mixture of essential oil, ripening inhibitor, and surfactant (Tween 80) into 5 mM sodium citrate buffer (pH 3.5). Stable nanoemulsions containing small droplets (d < 70 nm) were formed. The antimicrobial activity of the nanoemulsions decreased with increasing ripening inhibitor concentration which was attributed to a reduction in the amount of hydrophobic antimicrobial constituents transferred to the separated hydrophobic domain, mimicking bacterial cell membranes, by using dialysis and chromatography. The antimicrobial activity of the nanoemulsions also depended on the nature of the ripening inhibitor used: palm ≈ corn > canola > coconut which also depended on their ability to transfer hydrophobic antimicrobial constituents to the separated hydrophobic domain.