Sodium benzoate and sodium bisulfate as preservatives in apple juice and alternative sanitizers for washing cherry tomatoes.

Unpasteurized apple ciders and fresh produce have been linked to multistate outbreaks due to contamination by foodborne pathogens. Organic acids such as benzoic acid are effective antimicrobials, and acidified sodium benzoate (NaB) has been reported to be effective in reducing pathogens inoculated on cherry tomatoes and preventing cross-contamination. Sodium bisulfate (SBS) is a powerful acidulant but has not been studied in combination with NaB. The objective of the present study was to characterize the antibacterial activity of SBS and its combination with NaB in tryptic soy broth (TSB) and apple juice, as well as washing cherry tomatoes. The minimum inhibitory concentration and minimum bactericidal concentration of SBS were all 0.5% w/v (corresponding to TSB medium pH of 4.30) and 1.0% w/v (corresponding to TSB medium pH of 2.88), respectively, for Escherichia coli O157:H7 ATCC 43895, Salmonella Enteritidis ATCC 13076, and Listeria monocytogenes Scott A. In TSB, the triple combination of 1.0% w/v SBS, 0.1% w/v NaB, and 0.02% w/v oregano oil (OO) showed the faster inactivation rate of the three bacteria than treatments with one or two antimicrobials; the activity of double combinations followed the order of 0.1% w/v NaB +1.0% w/v SBS > 0.1% w/v NaB +0.5% w/v SBS + 0.02% w/v OO > 0.1% w/v NaB +0.5% w/v SBS. pH was a critical factor in the activity of antimicrobial combinations in TSB, and L. monocytogenes was more resistant than Gram-negative E. coli O157:H7 and S. Enteritis. In apple juice added with 0.05% w/v NaB, 0.25% w/v SBS, and 0.01% w/v OO alone or in combinations, 5 log CFU/mL or greater reductions in 72 h were observed for E. coli O157:H7 and S. Enteritidis in double and triple combinations, while only the triple combination and the SBS-OO combination resulted in the same effect for L. monocytogenes. For cherry tomatoes inoculated with 6.8 log CFU/g E. coli O157:H7, complete decontamination (>6 log CFU/g) was achieved after soaking for 1 min in solutions containing 0.5-1.5% w/v SBS and 0.1% w/v NaB or 1.5% w/v SBS alone, and no pathogens were detected in all wash solutions containing 0.5-1.5% w/v SBS with and without NaB. The lower pH of wash solutions with a higher amount of SBS was a dominant factor in decontamination and prevention of cross-contamination. The present study showed the potential of SBS and its combination with NaB to enhance the safety of apple juice and cherry tomatoes.

Physical and antimicrobial properties of self-emulsified nanoemulsions containing three synergistic essential oils.

The multiantibiotic resistant characteristic of Staphylococcus aureus and the prevalence of foodborne illnesses due to foods, particularly dairy products, contaminated by the pathogen call for alternative preservation technologies. Essential oils (EOs) have shown activities against various bacterial pathogens. However, EOs are water-insoluble and have unpleasant sensory properties, and strategies are needed to prepare colloidal systems for dispersion in liquid products and increase their antimicrobial activity and therefore reduce their usage level. In the present study, the overall objective was to fabricate and characterize self-emulsified nanoemulsions containing multiple EOs with synergistic antibacterial activities. Cinnamon, lemongrass, oregano, and tea tree EOs were observed to have synergistic antibacterial activities when used in combinations, especially for their triple combinations showing the fractional inhibitory concentration index of 0.625 and below. When triple EOs were emulsified by the combination of sunflower lecithin and Tween 20, mixing EOs with corn oil enabled the preparation of translucent oil-in-water nanoemulsions by simply stirring the mixtures. Three formulations were chosen for the preparation of self-emulsified nanoemulsions as these systems remained translucent, the droplet diameter remained to be smaller than 200 nm, and the polydispersity index remained to be smaller than 0.3 after 180-day ambient storage at 21 °C. The oil: surfactant mass ratio in these nanoemulsions (1.16 or higher) was higher than that of EO microemulsions (up to 0.77) prepared with a similar method. The minimum inhibitory concentration (MIC) of the nanoemulsions remained to be 0.5 or 1.0 mg/mL against S. aureus, while the minimum bactericidal concentration (MBC) remained to be 1.0 or 2.0 mg/mL after the 180-day storage. A higher EO: surfactant mass ratio of nanoemulsions resulted in a lower MIC and MBC. When tested in whole milk with 3.3% fat at 21 °C, no inhibition of S. aureus was observed for nanoemulsions at overall EO levels of 2.5 and 5.0 mg/mL; at an overall EO concentration of 7.5 mg/mL, no inhibition was observed for one nanoemulsion with the lowest EO:(corn oil + surfactant) mass ratio, while the gradual reduction of S. aureus by ~1 or 2 log CFU/mL in 120 h was observed for the other two nanoemulsions. The formulations and nanoemulsion preparation method in the present study may be significant to the utilization of EOs as natural preservatives to improve food safety.

Inactivation of Escherichia coli K12 on raw almonds using supercritical carbon dioxide and thyme oil.

Raw almonds could be contaminated by pathogens, but the current pasteurization practice using propylene oxide in the U.S. has flammability and carcinogenicity concerns. Supercritical carbon dioxide (scCO2) is a water-free technology and is a solvent of essential oils that are effective antimicrobial preservatives. The objective of this study was to investigate the possibility of combining scCO2 and thyme oil (TO) to reduce Escherichia coli K12 inoculated on raw almonds. Raw almonds inoculated with ∼6 log CFU/g E. coli K12 were batch-treated with scCO2 alone or the combination of presoaking in pure TO followed by scCO2 treatments at different combinations of temperature, pressure, and duration. Compared to scCO2 alone treatments, the combination of TO and scCO2 treatments significantly improved the disinfection effectiveness. Temperature had the most significant effect on the log reduction. At 70 °C, the log reduction by the combination treatment was over 4-log CFU/g and the maximum reduction was 5.16 log CFU/g. The findings suggest that the combination of TO and scCO2 may be a potential water-free technology to meet the requirement of over 4-log reduction of target microorganism for almond and other tree nut products.

Potential of acidified sodium benzoate as an alternative wash solution of cherry tomatoes: Changes of quality, background microbes, and inoculated pathogens during storage at 4 and 21°C post-washing.

To evaluate the feasibility of acidified sodium benzoate (NaB) as alternative washing solutions of fresh produce, the survival of inoculated pathogens, the background molds and yeasts counts, and quality parameters were compared during 4 and 21 °C storage of cherry tomatoes washed with 3000 ppm NaB at pH 2.0, 200 ppm free chlorine at pH 6.5, water adjusted to pH 2.0, and distilled water. The acidified NaB solution was the most effective in reducing the population of Escherichia coli O157:H7, Salmonella enterica and Listeria monocytogenes cocktails on tomatoes (>4 log CFU/g). NaB was more effective than free chlorine (P < 0.05) in reducing the two Gram-negative bacteria on tomatoes, while the reduction of Gram-positive L. monocytogenes by NaB (5.49 log CFU/g) and chlorine (4.98 log CFU/g) was similar (P > 0.05). No recovery of bacteria was found in all treatments during storage for 15 days. The acidified NaB effectively controlled yeasts and molds on cherry tomatoes to <1 log CFU/g or below the detection limit at both temperatures during 15-d storage, while free chlorine did not. Compared to unwashed controls, NaB had no effect on color, weight loss, firmness, and total soluble solids content of tomatoes during storage. The effect of NaB reducing pathogenic and spoilage microorganisms on tomatoes and maintaining quality during storage suggests its potential as an alternative wash solution in postharvest processing of fresh produce.

Antibacterial activity of acidified sodium benzoate against Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes in tryptic soy broth and on cherry tomatoes.

Concerns about undesirable by-products from chlorine sanitation of fresh produce and the limited efficacy with the presence of organic matter, have led to studies on alternative washing solutions. The aim of this study was to evaluate the antibacterial activities of acidified sodium benzoate (NaB) solutions against Escherichia coli O157:H7, Salmonella enterica and Listeria monocytogenes in growth medium and on cherry tomatoes. Experimentally, the minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs, >3 Log reduction) of NaB against E. coli O157:H7 ATCC 43895, S. Enteritidis, and L. monocytogenes Scott A were determined at pH 7.0-4.0 using micro-broth dilution method and agar plating method, respectively. The reduction of the three bacteria in tryptic soy broth (TSB) by 500 and 1000 ppm NaB at pH 2.0, 2.5 and 3.0 for 30 min at 21 °C was compared. Residual bacterial cocktails inoculated on cherry tomatoes were determined after soaking in 3000 ppm NaB solution adjusted to pH 2.0 for 3 min at 21 °C. Results showed that the MBC of NaB reduced from >10,000 ppm at pH 7.0 to 1000 ppm at pH 4.0 and was identical for the three bacteria. The log reduction of bacteria in TSB indicated that 1000 ppm NaB at pH 2.0 was the most effective in killing the three pathogens. The respective reduction of E. coli O157:H7 and S. enterica cocktails inoculated on cherry tomatoes immersed in 3000 ppm NaB (pH 2.0) at 21 °C for 3 min was 4.99 ±â€¯0.57 and 4.08 ±â€¯0.65 log CFU/g, which was significantly higher (p < 0.05) than the treatments of 200 ppm free chlorine at pH 6.5. Conversely, the reduction of L. monocytogenes on tomatoes by 3000 ppm NaB (4.88 ±â€¯0.73 log CFU/g) was similar (p > 0.05) to 200 ppm chlorine. Furthermore, the reduction of bacterial cocktails on tomatoes by 3000 ppm NaB at pH 2.0 was not affected after adding 1% tomato puree, and bacteria were not detected in NaB washing solutions with and without 1% tomato puree and on following un-inoculated tomatoes. This study showed that acidified NaB solution may be used as an alternative post-harvest wash of produce.

Self-assembled curcumin-soluble soybean polysaccharide nanoparticles: Physicochemical properties and in vitro anti-proliferation activity against cancer cells.

Nanoencapsulation of lipophilic bioactive compounds in food biopolymers is important to functional beverages, but protein-based nanocapsules are unstable around the isoelectric point of protein. The objectives of this work were to study physicochemical properties of self-assembled curcumin-soluble soybean polysaccharide (SSPS) nanoparticles and evaluate the activities against proliferation of human colon HCT116 and mammary adenocarcinoma MCF-7 cancer cells before and after simulated digestions. Capsules with a hydrodynamic diameter of 200-300 nm and an encapsulation efficiency of ∼90% were self-assembled after increasing curcumin-SSPS mixture to pH 12.0 and lowering pH to 7.0. The capsule dispersions were stable at pH 2.0-7.0 and after heating at 95 °C for 1 min. No significant difference was observed for the viability of HCT 116 and MCF-7 cells challenged with 0.4, 4.0, and 40 µg/ml nanoencapsulated curcumin before and after simulated gastric and intestinal digestions. These findings may be significant to help develop functional beverages for disease prevention.

Lactobionic acid enhances the synergistic effect of nisin and thymol against Listeria monocytogenes Scott A in tryptic soy broth and milk.

Listeria monocytogenes is a Gram-positive opportunistic human pathogen and it remains a significant cause of foodborne illnesses. A variety of natural and synthetic compounds have been studied to inhibit the growth of L. monocytogenes in foods. Antimicrobial combinations with synergistic antilisterial properties can reduce the dose of each antimicrobial, which can be further enhanced by chelating compounds. Therefore, the objective of this study was to determine antilisterial properties of binary or ternary combinations of lactobionic acid (LBA), nisin, and thymol in tryptic soy broth (TSB), 2% reduced-fat milk, and whole milk. The results showed that the minimum inhibitory concentration (MIC) of nisin, thymol and LBA was 125IU/mL, 0.25mg/mL, and 10mg/mL, respectively. The ternary combination was the most effective in reducing MICs of antimicrobials, with the MIC of nisin, thymol, and LBA being 31.25IU/mL, 0.0625mg/mL, and 1.25mg/mL, respectively. In TSB with 0.6% yeast extract, L. monocytogenes grew in individual or binary antimicrobial treatments of 31.25IU/mL nisin, 0.0625mg/mL thymol, and 1.25mg/mL LBA within 24h at 32°C, while it was completely inhibited by the ternary combination. In 2% reduced-fat milk at 21°C, the ternary combination of nisin, thymol, and LBA at respective concentrations of 250IU/mL, 2mg/mL, and 10mg/mL completely inhibited the bacterium to below the detection limit in 72h while >2log (CFU/mL) bacteria was still detected in all the binary combinations after 120h. In whole milk, the combination of 500IU/mL nisin, 2mg/mL thymol, and 10mg/mL LBA reduced bacteria to around 2log (CFU/mL) in 4h at 21°C, and no bacterial recovery was observed after 5 d. This study suggested the potential of the ternary combination of nisin, thymol and LBA for food preservation.

Potential of Cinnamon Oil Emulsions as Alternative Washing Solutions of Carrots.

The objective of this study was to evaluate the potential of cinnamon oil emulsions as alternative washing solutions to improve the microbial safety of carrots. Whey protein concentrate (WPC), gum arabic (GA), lecithin, and their combinations were used to prepare cinnamon oil emulsions. The emulsions were characterized for their hydrodynamic diameter (Dh) during 7 days of storage and their antimicrobial activity against cocktails of Salmonella enterica , Escherichia coli O157:H7, and Listeria monocytogenes . The Dh of the emulsion prepared with the GA+WPC blend did not change significantly (195.0 to 184.1 nm), whereas all other emulsions showed varying degrees of increases in Dh. Compared with free cinnamon oil dissolved in 5% ethanol, all emulsions showed similar or lower MICs and MBCs. Emulsions prepared with GA and equal masses of GA and WPC were chosen and diluted to 0.2 and 0.5% cinnamon oil to wash carrots that were surface inoculated with bacterial cocktails because of their lower MICs and MBCs than free oil. Emulsions resulted in significantly higher reductions of pathogens on carrots than free cinnamon oil, 3.0 to 3.7 versus 2.1 to 2.3 log CFU/g at 0.5% cinnamon oil and 2.0 to 3.0 versus 1.0 to 1.7 log CFU/g at 0.2% cinnamon oil. No transfer of bacteria from inoculated carrots to wash solutions and no effects of organic load on log reductions were only observed for wash treatments with 0.5% emulsified cinnamon oil. Thus, the cinnamon oil emulsions are potential alternative postharvest washing solutions for fresh produce production.

Microemulsions based on a sunflower lecithin-Tween 20 blend have high capacity for dissolving peppermint oil and stabilizing coenzyme Q10.

The objectives of the present study were to improve the capability of microemulsions to dissolve peppermint oil by blending sunflower lecithin with Tween 20 and to study the possibility of codelivering lipophilic bioactive compounds. The oil loading in microemulsions with 20% (w/w) Tween 20 increased from 3% (w/w) to 20% (w/w) upon gradual supplementation of 6% (w/w) lecithin. All microemulsions had particles of <12 nm that did not change over 70 d of storage at 21 °C. They had relatively low Newtonian viscosities and were physically and chemically stable after 50-200-fold dilution in water, resulting from similar hydrophile-lipophile-balance values of the surfactant mixture and peppermint oil. Furthermore, the microemulsions were capable of dissolving coenzyme Q10 and preventing its degradation at UV 302 nm, more significant for the microemulsion with lecithin. Therefore, natural surfactant lecithin can reduce the use of synthetic Tween 20 to dissolve peppermint oil and protect the degradation of dissolved lipophilic bioactive components in transparent products.

Thermal and UV stability of ß-carotene dissolved in peppermint oil microemulsified by sunflower lecithin and Tween 20 blend.

Microemulsions are suitable for simultaneous delivery of flavour oils and lipophilic bioactive compounds in transparent beverages. In the present study, the feasibility of delivering ß-carotene in microemulsions formulated with peppermint oil and a blend of Tween® 20 and various amounts of sunflower lecithin was investigated. The poorly water- and oil-soluble ß-carotene was dissolved in the transparent microemulsions that had particles smaller than 10nm and were stable during ambient storage for 65 d. The inclusion of ß-carotene did not change the flow-behaviour and Newtonian viscosity. The degradation of ß-carotene in microemulsions during ambient storage, ultraviolet radiation, and thermal treatments at 60 and 80 °C followed first order kinetics and was greatly suppressed when compared to the solution control. The antioxidant potential of peppermint oil and a greater content of lecithin in microemulsions enabled the better protection of ß-carotene. The studied microemulsions may find various applications in manufacturing transparent beverages.

Antimicrobial properties of nisin after glycation with lactose, maltodextrin and dextran and the thyme oil emulsions prepared thereof.

To clarify the reported conflicting antimicrobial activities of nisin after glycation, nisin was glycated with lactose, maltodextrin, and dextran at 70 °C and 50% relative humidity for 1-24 h. Nisin before and after glycation was studied for the first time to prepare thyme oil emulsions. The activity of glycated nisin and the thyme oil emulsions was tested in both tryptic soy broth (TSB) and 2% reduced fat milk. Results showed that nisin glycated with a smaller saccharide for a longer duration had a higher degree of glycation and the reduced number of positive charges lowered its antibacterial activity. The emulsified thyme oil had an additive effect with either glycated or native nisin against Listeria monocytogenes Scott A and Bacillus subtilis in TSB and 2% reduced fat milk. However, emulsions were less effective against L. monocytogenes Scott A in milk than same units of native nisin and same concentration of free thyme oil, likely due to the reduced availability of thymol and carvacrol, the main components of thyme oil. These results showed that glycation of nisin cannot broaden its antimicrobial activity and nisin is not a good compound to prepare emulsions of essential oils.

Thymol nanoencapsulated by sodium caseinate: physical and antilisterial properties.

In this work, thymol was encapsulated in sodium caseinate using high shear homogenization. The transparent dispersion at neutral pH was stable for 30 days at room temperature as determined by dynamic light scattering and atomic force microscopy, which agreed with high ζ potential of nanoparticles. The slightly decreased particle dimension during storage indicates the absence of Ostwald ripening. When molecular binding was studied by fluorescence spectroscopy, thymol was observed to bind with tyrosine and possibly other amino acid residues away from tryptophan of caseins. At pH 4.6 (isoelectric point of caseins), the stabilization of thymol nanoparticles against aggregation was enabled by soluble soybean polysaccharide, resulting from the combined electrostatic and steric repulsions. The encapsulated thymol showed the significantly improved antilisterial activity in milk with different fat levels when compared to thymol crystals, resulting from the quicker mixing and increased solubility in the milk serum. The transparent thymol nanodispersions have promising applications to improve microbiological safety and quality of foods.

Impacts of sample preparation methods on solubility and antilisterial characteristics of essential oil components in milk.

Essential oil components (EOCs) have limited water solubility and are used at much higher concentrations in complex food matrices than in growth media to inhibit pathogens. However, the correlation between solubility and activity has not been studied. The objective of this work was to characterize the solubility of EOCs in solvents and milk and correlate solubility with antilisterial activity. The solubilities of four EOCs, thymol, carvacrol, eugenol, and trans-cinnamaldehyde, in water was significantly increased in the presence of 5% (vol/vol) ethanol. In milk, the solubility of EOCs was lower than in water, with lower solubility in higher-fat milk. EOCs applied to milk as stock solutions (in 95% aqueous ethanol) enabled quicker dissolution and higher solubility in milk serum than other methods of mixing, such as end to end, and greater reductions of Listeria monocytogenes Scott A after 0 and 24 h. When the EOC concentration detected in milk serum was above the minimum bactericidal concentration, complete inhibition of L. monocytogenes in tryptic soy broth resulted. Therefore, the antilisterial properties in milk could be correlated with the solubility by comparison to the minimum inhibitory or bactericidal concentrations of EOCs. While the EOCs applied using ethanol generally had solubility and activity characteristics superior to those of other mixing methods, ethanol is not used to a great extent in nonfermented foods. Therefore, mixing methods without an organic solvent may be more readily adaptable to enhancing the distribution of EOCs in complex food systems.

Whole-tree agarwood-inducing technique: an efficient novel technique for producing high-quality agarwood in cultivated Aquilaria sinensis trees.

Agarwood is the fragrant resin-infused wood derived from the wounded trees of Aquilaria species. It is a valuable non-timber forest product used in fragrances and as medicine. Reforestation for Aquilaria trees in combination with artificial agarwood-inducing methods serves as a way to supply agarwood and conserve of wild Aquilaria stock. However, the existing agarwood-inducing methods produce poor-quality agarwood at low yield. Our study evaluated a novel technique for producing agarwood in cultivated Aquilaria trees, called the whole-tree agarwood-inducing technique (Agar-Wit). Ten different agarwood inducers were used for comparison of Agar-Wit with three existing agarwood-inducing methods. For Aquilaria trees treated with these ten inducers, agarwood formed and spread throughout the entire tree from the transfusion point in the trunk to the roots and branches of the whole tree. Agarwood yield per tree reached 2,444.83 to 5,860.74 g, which is 4 to 28 times higher than that by the existing agarwood-inducing methods. Furthermore, this agarwood derived from Agar-Wit induction was found to have a higher quality compared with the existing methods, and similar to that of wild agarwood. This indicates Agar-Wit may have commercial potential. Induction of cultivated agarwood using this method could satisfy the significant demand for agarwood, while conserving and protecting the remaining wild Aquilaria trees.

Comparison of compositions and antimicrobial activities of essential oils from chemically stimulated agarwood, wild agarwood and healthy Aquilaria sinensis (Lour.) gilg trees.

The composition and antimicrobial activity of the essential oils which were obtained from agarwood originated from Aquilaria sinensis (Lour.) Gilg stimulated by the chemical method (S1) were characterized, taking wild agarwood (S2) and healthy trees (S3) respectively as the positive and negative controls. The chemical composition of S1 was investigated by gas chromatography-mass spectrometry (GC-MS). The essential oil of S1 showed a similar composition to that of S2, being rich in sesquiterpenes and aromatic constituents. However, the essential oil of S3 was abundant in fatty acids and alkanes. Essential oils of S1 and S2 had better inhibition activities towards Bacillus subtilis and Staphyloccus aureus, compared with essential oil of S3. Escherichia coli was not sensitive to any of them.