Effect of anthocyanins on lipid oxidation and microbial spoilage in value-added emulsions with bilberry seed oil, anthocyanins and cold set whey protein hydrogels.

The objective of this work was to explore the storage properties of a structured oil-in-water emulsion containing both water- and fat-soluble bioactive compounds from bilberries (Vaccinium myrtillus L.). Bilberry seed oil (BSO) was dispersed in a continuous aqueous phase of anthocyanins (AC) and whey protein isolate. The microstructure was evaluated using light microscopy and the effect of anthocyanins on lipid oxidation and microbial growth was investigated. The results showed that it was possible to generate a stable emulsion structure that resisted phase separation during 25 weeks of storage. Gas chromatography-mass spectrometry measurements of the fatty acids in the BSO during storage showed that AC had a protective effect against lipid oxidation. The AC did not have an antimicrobial effect against the investigated strains Zygosaccharomyces bailii (ATCC 42476) and Aspergillus niger (ATCC 6275 (M68)).

Preservative effect of Camellia sinensis (L.) Kuntze seed extract in soy sauce and its mutagenicity.

The purpose of this study was to investigate the applicability of green tea seed (GTS) extract as a natural preservative in food. Food preservative ability and mutagenicity studies of GTS extract and identification of antimicrobial compounds from GTS extract were carried out. The GTS extract showed only anti-yeast activity against Candida albicans with MIC value of 938µg/mL and Zygosaccharomyces rouxii with a MIC of 469µg/mL. The active compounds were identified as theasaponin E1 (1), assamsaponin A (2), and assamsaponin B (3). And GTS extracts didn't show mutagenicity because there were no dose-dependent changes in colonies of Salmonella typhimurium TA98, TA100, TA1535, TA1537, and Escherichia coli WP2uvrA regardless of the metabolic activation system. And GTS extract also showed a potent food preservation affect which eliminated all yeast below the MIC value in application test at soy sauce. Overall, these results indicate that GTS extract could be a safe and effective food preservative with anti-yeast activity.

Characterization of in vitro antifungal activities of small and American cranberry (Vaccinium oxycoccos L. and V. macrocarpon Aiton) and lingonberry (Vaccinium vitis-idaea L.) concentrates in sugar reduced fruit spreads.

In this study, cranberry and lingonberry concentrates were added to commercial sugar-reduced fruit spreads (raspberry-Aloe vera, strawberry-guava, and strawberry-lime), and tested for their antifungal activities. Selected strains of the species Absidia glauca, Penicillium brevicompactum, Saccharomyces cerevisiae and Zygosaccharomyces bailii, as well as xerophilic environmental isolates of the genera Penicillium and Eurotium were used for challenge testing. Initially, varying concentrations of synthetic antifungal agents, such as sodium benzoate, potassium sorbate and butyl 4-hydroxybenzoate were tested against these fungi on wort agar containing 31% fructose at different pH values. Subsequently, the experiments were conducted in fruit spreads containing different concentrations of cranberry and lingonberry concentrates. The results of this study demonstrate that these concentrates were able to inhibit growth of visible colonies of xerophilic and non-xerophilic fungi. Cranberry and lingonberry concentrates are interesting candidates for natural preservation against fungal growth in sugar reduced fruit spreads.

Prenylated flavonoids from the stems and leaves of Desmodium caudatum and evaluation of their inhibitory activity against the film-forming growth of Zygosaccharomyces rouxii F51.

In order to provide scientific evidence for the relationship between the traditional usage, stems and leaves of Desmodium caudatum being used for protecting miso from spoilage, and its Japanese name (miso-naoshi), phytochemical study on the stems and leaves of this plant was carried out. Seven new prenylated flavonoids (1-3, 15-18), together with 19 known compounds (4-14, 19-26), were isolated, and the structures of new compounds were elucidated by extensive spectroscopic analyses. The minimum inhibitory concentrations (MICs) of 28 flavonoids, including 17 compounds (1, 2, 4, 5, 7-14, 20-22, 24, 25) isolated in this study and 11 flavonoids (27-37) previously isolated from the roots of this plant, against the film-forming yeast of Zygosaccharomyces rouxii F51 were determined. Fifteen compounds (2, 4, 5, 11, 12, 14, 21, 22, 25, 27, 28, 32-35) inhibited the film-forming growth of Z. rouxii F51 (MIC values, 7.8-62.5 µg/mL), among which 2",2"-dimethylpyran-(5",6":7,8)-5,2'-dihydroxy-4'-methoxy-(2R,3R)-dihydroflavonol (11) demonstrated potent inhibitory activity with an MIC value of 7.8 µg/mL.

Suitability of Bifidobacterium spp. and Lactobacillus plantarum as probiotics intended for fruit juices containing citrus extracts.

A strain of Lactobacillus plantarum and 4 strains of bifidobacteria were inoculated in apple juice and in a commercial beverage labeled as "red-fruit juice," containing citrus extracts as natural preservatives; the suitability of the probiotics was evaluated in relation to their resistance to 2 kinds of citrus extracts (biocitro and lemon extract), survival in juices at 4 and 37 °C, and inhibition of Zygosaccharomyces bailii. Cell count of L. plantarum and bifidobacteria over time was fitted through the Weibull equation, for the evaluation of the first reduction time (δ), death time, and microbiological shelf life (the break-point was set to 7 log cfu/mL). Bifidobacterium animalis subsp. lactis experienced the highest δ-value (23.21 d) and death time (96.59 d) in the red-fruit juice at 4 °C, whereas L. plantarum was the most promising strain in apple juice at 37 °C. Biocitro and lemon extract did not exert a biocidal effect toward probiotics; moreover, the probiotics controlled the growth of Z. bailii and the combination of L. plantarum with 40 ppm of biocitro reduced the level of the yeast after 18 d by 2 log cfu/mL.

Inhibition of spoiling yeasts of fruit juices through citrus extracts.

This article reports on the bioactivities of citrus extracts (citrus extract, lemon extract, and neroli) toward Saccharomyces cerevisiae, Zygosaccharomyces bailii, Zygosaccharomyces rouxii, Pichia membranifaciens, and Rhodotorula bacarum. The bioactivities of the extracts (from 10 to 100 ppm) were evaluated through a microdilution method; thereafter, citrus extracts (0 to 80 ppm) were tested in combination with either pH (3.0 to 5.0) or temperature (5 to 25°C). Finally, a confirmatory experiment was run in a commercial drink (referred to as red fruit juice) containing citrus extract (40 ppm) that was inoculated with either S. cerevisiae or Z. bailii (5 log CFU/ml) and stored at 4 and 25°C. Yeasts increased to 7 log CFU/ml (Z. bailii) or 8 log CFU/ml (S. cerevisiae) in the control at 25°C, but the citrus extract addition controlled yeast growth for at least 3 days; under refrigeration, the effect was significant for 10 days.

Physical properties and antimicrobial efficacy of thyme oil nanoemulsions: influence of ripening inhibitors.

Thyme oil-in-water nanoemulsions (pH 3.5) were prepared as potential antimicrobial delivery systems. The nanoemulsions were highly unstable to droplet growth and phase separation, which was attributed to Ostwald ripening due to the relatively high water solubility of thyme oil. Ostwald ripening could be inhibited by mixing thyme oil with a water-insoluble ripening inhibitor (≥60 wt % corn oil or ≥50 wt % MCT in the lipid phase) before homogenization, yielding nanoemulsions with good physical stability. Physically stable thyme oil nanoemulsions were examined for their antimicrobial activities against an acid-resistant spoilage yeast, Zygosaccharomyces bailii (ZB). Oil phase composition (ripening inhibitor type and concentration) had an appreciable influence on the antimicrobial activity of the thyme oil nanoemulsions. In general, increasing the ripening inhibitor levels in the lipid phase reduced the antimicrobial efficacy of nanoemulsions. For example, for nanoemulsions containing 60 wt % corn oil in the lipid phase, the minimum inhibitory concentration (MIC) of thyme oil to inhibit ZB growth was 375 µg/mL, while for nanoemulsions containing 90 wt % corn oil in the lipid phase, even 6000 µg/mL thyme oil could not inhibit ZB growth. This effect is also dependent on ripening inhibitor types: at the same concentration in the lipid phase, MCT decreased the antimicrobial efficacy of thyme oil more than corn oil. For instance, when the level of ripening inhibitor in the lipid phase was 70 wt %, the MICs of thyme oil for nanoemulsions containing corn oil and MCT were 750 and 3000 µg/mL, respectively. The results of this study have important implications for the design and utilization of nanoemulsions as antimicrobial delivery systems in the food and other industries.

Application of proanthocyanidins from peanut skins as a natural yeast inhibitory agent.

UNLABELLED: Proanthocyanidins were extracted from peanut skins and investigated for their antimicrobial activity against Saccharomyces cerevisiae, Zygosaccharomyces bailii, and Zygosaccharomyces bisporus in traditional growth media (Sabouraud Dextrose and Maltose broth) and a simulated apple juice beverage. Peanut skins extracts (PSE) were prepared through a multisolvent extraction procedure. The PSE extended the lag phase growth of the 3 yeasts studied at a concentration of 1 mg/mL and at 10 mg/mL yeast growth was totally inhibited for 120 h. PSE was fractionated by normal phase high performance liquid chromatography and the active components/fractions were determined. Compounds present in the fractions were identified by liquid chromatography-mass spectrometry to determine the compounds responsible for inhibition. Fractions consisting mostly of A-type proanthocyanidin dimers, trimers, and tetramers showed the highest percent inhibition toward the yeasts tested in this study. Both optical density (OD) and standard enumeration plating methods were performed in this study. The OD method led to an overestimation of the inhibitory effects of PSE, the 2 methods agreed in respect to treatment effects but not the severity of the inhibition. PRACTICAL APPLICATION: There is a growing consumer demand for "fresh like" products containing reduced amounts of chemical preservatives without compromising food safety and quality. Therefore, the goal of this study was to determine if an extract of peanut skins containing flavonoid rich compounds could function as a natural antimicrobial in a model beverage system. Proteins were removed through the process of producing the peanut skin extract, thus it is unlikely to contain peanut allergens. The antimicrobial compounds mentioned in this study were successfully integrated into a model beverage system, and were found to have antimicrobial effect. However, the incorporation of these compounds would likely lead to negative sensory attributes at the concentration needed to achieve an appreciable antimicrobial effect alone.

Cationic antimicrobial (ε-polylysine)-anionic polysaccharide (pectin) interactions: influence of polymer charge on physical stability and antimicrobial efficacy.

The cationic biopolymer ε-polylysine (ε-PL) is a potent food-grade antimicrobial that is highly effective against a range of food pathogens and spoilage organisms. In compositionally complex systems such as foods and beverages, cationic ε-PL molecules may associate with anionic substances, leading to increased turbidity, sediment formation, and reduced antimicrobial activity. This study therefore characterized the interactions between cationic ε-PL and anionic pectins with different degrees of esterification (DE) and then investigated the influence of these interactions on the antimicrobial efficacy of ε-PL. The nature of the interactions was characterized using isothermal titration calorimetry (ITC), microelectrophoresis (ME), and turbidity measurements. High (DE 61%), medium (DE 51%), and low (DE 42%) methoxyl pectins interacted with ε-PL molecules through electrostatic forces, forming either soluble or insoluble complexes with various electrical charges, depending on the relative mass ratio of pectin and ε-PL. The interaction of pectin with ε-PL increased as the negative charge density on the pectin molecules increased, that is, with decreasing DE. The antimicrobial efficacy of ε-PL against two acid-resistant spoilage yeasts (Zygosaccharomyces bailii and Saccharomyces cerevisiae) decreased progressively in the presence of increasing levels of all three pectins. Nevertheless, the low DE pectin decreased the antimicrobial efficacy of ε-PL much more dramatically, likely due to strong electrostatic binding of ε-PL onto low DE pectin molecules reducing its interaction with anionic microbe surfaces. This study provides knowledge that will facilitate the rational application of ε-PL as an antimicrobial in complex food systems.

Antifungal activity of alkanols against Zygosaccharomyces bailii and their effects on fungal plasma membrane.

A series of aliphatic primary alkanols from C(6) to C(13) were tested for antifungal activity against a food spoilage fungus Zygosaccharomyces bailii using a broth dilution method and were compared for their effects against Saccharomyces cerevisiae and Z. rouxii. Decanol (C(10)) was found to be the most potent fungicide against Z. bailii at a minimum fungicidal concentration of 50 microg/ml (0.31 mM), whereas undecanol (C(11)) was found to be the most potent fungistatic at a minimum inhibitory concentration of 25 microg/ml (0.14 mM). The time-kill curve study showed that decanol was fungicidal against Z. bailii at any growth stage. Octanol (C(8)) increased plasma membrane fluidity in the spheroplast cells of S. cerevisiae. The primary antifungal action of alkanols comes from their ability to disrupt the native membrane-associated function of integral proteins nonspecifically as nonionic surface-active agents (surfactants). The antifungal activity of decanol against Z. bailii was slightly enhanced in combination with anethole.

Caffeine induces macroautophagy and confers a cytocidal effect on food spoilage yeast in combination with benzoic acid.

Weak organic acids are an important class of food preservatives that are particularly efficacious towards yeast and fungal spoilage. While acids with small aliphatic chains appear to function by acidification of the cytosol and are required at high concentrations to inhibit growth, more hydrophobic organic acids such as sorbic and benzoic acid have been suggested to function by perturbing membrane dynamics and are growth-inhibitory at much lower concentrations. We previously demonstrated that benzoic acid has selective effects on membrane trafficking in Saccharomyces cerevisiae. Benzoic acid selectively blocks macroautophagy in S. cerevisiae while acetic acid does not, and sorbic acid does so to a lesser extent. Indeed, while both benzoic acid and nitrogen starvation are cytostatic when assayed separately, the combination of these treatments is cytocidal, because macroautophagy is essential for survival during nitrogen starvation. In this report, we demonstrate that Zygosaccharomyces bailii, a food spoilage yeast with relatively high resistance to weak acid stress, also exhibits a cytocidal response to the combination of benzoic acid and nitrogen starvation. In addition, we show that nitrogen starvation can be replaced by caffeine supplementation. Caffeine induces a starvation response that includes the induction of macroautophagy, and the combination of caffeine and benzoic acid is cytocidal, as predicted from the nitrogen starvation data.

Inhibitory activity of essential oils of garlic and onion against bacteria and yeasts.

The essential oils of garlic and onion and their constituent sulfides with three or more sulfur atoms were potent inhibitors of yeast growth. The minimum inhibitory concentrations of garlic oil, onion oil, diallyl trisulfide, diallyl tetrasulfide, and dimethyl trisulfide for all the yeasts tested ranged between 2 and 45 ppm. The oils and their constituent sulfides, however, were only very weakly antibacterial, showing minimum inhibitory concentrations of greater than 300 ppm for most of the bacteria tested. The antiyeast activity of garlic oil and onion oil was storage stable and was not influenced by pH. Film formation on soy sauce by Zygosaccharomyces rouxii SS1 was completely prevented for 30 days by the addition of 30 and 40 ppm of garlic oil and onion oil, respectively.

The antimicrobial properties of chitosan in mayonnaise and mayonnaise-based shrimp salads.

The potential for using chitosan glutamate as a natural food preservative in mayonnaise and mayonnaise-based shrimp salad was investigated. Mayonnaise containing 3 g/liter of chitosan combined with acetic acid (0.16%) or lemon juice (1.2 and 2.6%) was inoculated with log 5 to 6 CFU/g of Salmonella Enteritidis, Zygosaccharomyces bailii, or Lactobacillus fructivorans and stored at 5 and 25 degrees C for 8 days. In mayonnaise containing chitosan and 0.16% acetic acid, 5 log CFU/g of L. fructivorans were inactivated, and numbers remained below the sensitivity limit of the plate counting technique for the duration of the experiment. Z. bailii counts were also reduced by approximately 1 to 2 log CFU/g within the first day of incubation at 25 degrees C, but this was followed by growth on subsequent days, giving an overall growth delay of 2 days. No differences in counts of Z. bailii in mayonnaise stored at 5 degrees C or of Salmonella Enteritidis stored at either temperature were observed. In mayonnaise containing lemon juice at both 1.2 and 2.6%, no substantial differences were observed between the controls and the samples containing chitosan. In shrimp salads stored at 5 degrees C, the presence of a coating of chitosan (9 mg/g of shrimp) inhibited growth of the spoilage flora from approximately log 8 CFU/g in the controls to log 4 CFU/g throughout 4 weeks. However, at 25 degrees C, chitosan was ineffective as a preservative. The results demonstrated that chitosan may be useful as a preservative when combined with acetic acid and chill storage in specific food applications.