Influence of fat content and water activity on the heating pattern of model systems submitted to microwave heating.

This research analyzed the influence of fat content (1%, 3%, 5%, and 10%, w.b.) and of water activity (0.85, 0.90, 0.95, and 0.99) on the effectiveness of microwave heating within model systems (agar gel). Findings indicate that an increased fat content promotes more uniform heating in the model systems and results in longer heating times to reach the target temperature of 50°C. In the model systems with different aw values, the warming of the central model zone was slower while in the corners, heating was accelerated. Additionally, in these models, heating times increased as the aw was decreased. PRACTICAL APPLICATION: The study of model systems allows for a detailed assessment of food when subjected to microwaves; and how heating is influenced by food properties such as aw and fat content. Findings indicate that changes in product formulation could improve the heating of ready-to-eat foods, either by increasing the fat content (better uniformity) or having high aw (shorter heating time).

Essential oils microemulsions prepared with high-frequency ultrasound: physical properties and antimicrobial activity.

Essential oils (EOs) have demonstrated antimicrobial activity against bacteria due to the effects of their major components. The direct application of EOs may present a rapid volatilization of its components and can decrease their effectiveness. Encapsulation by means of emulsification can provide protection to lipid compounds on a microscale. The aim of this study was to characterize microemulsions of cinnamon essential oil (CEO), oregano essential oil (OEO), and rosemary essential oil (REO) prepared by high-frequency ultrasound and evaluate their antimicrobial activities against Escherichia coli and Listeria monocytogenes. The microemulsions (oil-in-water, O/W) of EOs were prepared using high-frequency ultrasound, applying a wave amplitude of 84 µm for 15 min (REO and CEO) or 30 min (OEO). The antimicrobial activity was determined by inoculating 108 CFU/mL of bacteria. Nonsurvival of the bacteria was confirmed by plate count in tryptic soy agar, determining the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC). The microemulsions exhibited droplet size diameters of 1.98 to 5.46 µm, showing high encapsulation efficiencies (79.91-81.97%) and low separation rates (2.50-6.67%). The MIC and MBC for the microemulsions for both bacteria were 20-75% less than values obtained for the non-encapsulated EOs. This study demonstrates that high-frequency ultrasound is a suitable technique for obtaining stable microemulsions to deliver natural antimicrobials that can be applied to control bacteria of high relevance in food safety.

Antimicrobial activity of protein-containing fractions isolated from Lactobacillus plantarum NRRL B-4496 culture.

The interest in lactic acid bacteria, including Lactobacillus plantarum NRRL B-4496, has increased in recent years as bio-preservatives, due to the production of secondary metabolites capable of inhibiting pathogenic bacteria. The objectives of this study were to evaluate the antimicrobial activity, cytotoxicity and the anti-inflammatory response of L. plantarum NRRL B-4496 cell-free supernatant (CFS). Furthermore, the CFS was fractionated by size exclusion chromatography using Sephadex G-25, and a minimal inhibitory volume test was determined against a panel of pathogenic bacteria. The cytotoxicity and the inflammatory activities of the fractions were evaluated using the human-derived THP-1 cell line. Results of this study indicates that CFS of L. plantarum NRRL B-4496 possesses antimicrobial protein compounds against the pathogen Listeria monocytogenes and showed no toxicity nor a pro-inflammatory response to human macrophages. The obtained results contribute to the development of novel bio-preservatives, L. plantarum cell-free supernatant or its fractions, with a potential use in the food industry.

Microencapsulated feruloyl esterase-producing lactobacilli ameliorate lipid profile and glycaemia in high fat diet-induced obese mice.

The effect of oral administration of spray-dried microcapsules of feruloyl esterase (FE) producing Lactobacillus fermentum CRL1446 (Lf) and Lactobacillus johnsonii CRL1231 (Lj) on high fat diet-induced obese mice was investigated to evaluate whether these strains could be used as a biotherapeutic for obesity. Swiss albino mice were divided into a normal diet fed group receiving empty microcapsules (control), a high fat diet plus empty microcapsules (HFD group), HFD plus microcapsules with Lf (HFD-Lf group) and HDF plus microcapsules with Lj (HFD-Lj group). Microcapsules containing Lf or Lj at a dose of ~107 cells/day/mouse were given orally for 7 weeks. Body weight gain, adiposity index, plasma leptin, lipid profiles, glycaemia, insulinemia, oral glucose tolerance, intestinal FE, glutathione peroxidase and glutathione reductase (GR) activities were determined. Administration of lactobacilli (HFD-Lf and HFD-Lj groups) improved metabolic parameters (triglyceride, total cholesterol, low-density lipoprotein cholesterol levels) and cardiovascular risk indicators (37-46% decrease of atherogenic index), and reduced body weight gain (29-38%), adiposity index (42-62%), plasma leptin levels, liver weight and fat deposition in liver. Intestinal FE activities significantly increased in HFD-Lf (62%) and HFD-Lj group (48%), thus improving hepatic GR activity (42% increment) compared to HFD group. Moreover, L. johnsonii increased HDL-cholesterol and L. fermentum reduced blood glucose to levels similar to the control. These FE-producing lactobacilli have the potential to improve biomarkers involved in obesity by increasing intestinal FE activity.

Probiotic viability and storage stability of yogurts and fermented milks prepared with several mixtures of lactic acid bacteria.

Currently, the food industry wants to expand the range of probiotic yogurts but each probiotic bacteria offers different and specific health benefits. Little information exists on the influence of probiotic strains on physicochemical properties and sensory characteristics of yogurts and fermented milks. Six probiotic yogurts or fermented milks and 1 control yogurt were prepared, and we evaluated several physicochemical properties (pH, titratable acidity, texture, color, and syneresis), microbial viability of starter cultures (Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus) and probiotics (Lactobacillus acidophilus, Lactobacillus casei, and Lactobacillus reuteri) during fermentation and storage (35 d at 5°C), as well as sensory preference among them. Decreases in pH (0.17 to 0.50 units) and increases in titratable acidity (0.09 to 0.29%) were observed during storage. Only the yogurt with S. thermophilus, L. delbrueckii ssp. bulgaricus, and L. reuteri differed in firmness. No differences in adhesiveness were determined among the tested yogurts, fermented milks, and the control. Syneresis was in the range of 45 to 58%. No changes in color during storage were observed and no color differences were detected among the evaluated fermented milk products. Counts of S. thermophilus decreased from 1.8 to 3.5 log during storage. Counts of L. delbrueckii ssp. bulgaricus also decreased in probiotic yogurts and varied from 30 to 50% of initial population. Probiotic bacteria also lost viability throughout storage, although the 3 probiotic fermented milks maintained counts ≥ 10(7)cfu/mL for 3 wk. Probiotic bacteria had variable viability in yogurts, maintaining counts of L. acidophilus ≥ 10(7) cfu/mL for 35 d, of L. casei for 7d, and of L. reuteri for 14 d. We found no significant sensory preference among the 6 probiotic yogurts and fermented milks or the control. However, the yogurt and fermented milk made with L. casei were better accepted. This study presents relevant information on physicochemical, sensory, and microbial properties of probiotic yogurts and fermented milks, which could guide the dairy industry in developing new probiotic products.

Antifungal activity of lactobacilli and its relationship with 3-phenyllactic acid production.

In this study, 13 lactic acid bacteria (LAB) strains (including 5 Lactobacillus casei, 2 Lactobacillus rhamnosus, 2 Lactobacillus fermentum, 1 Lactobacillus acidophilus, 1 Lactobacillus plantarum, 1 Lactobacillus sakei, and 1 Lactobacillus reuteri species) were assessed for both their antifungal activity against four food spoilage molds (Colletotrichum gloeosporioides, Botrytis cinerea, Penicillium expansum, and Aspergillus flavus) and their capability to produce the novel antimicrobial compound 3-phenyllactic acid (PLA). Results demonstrated that all molds were sensitive to varying degrees to the cell-free supernatants (CFS) from LAB fermentations (p<0.05), with growth inhibitions ranging from 2.65% to 66.82%. The inhibition ability of CFS was not affected by a heating treatment (121°C, 20 min); however, it declined markedly when the pH of CFS was adjusted to 6.5. With the exception of L. plantarum NRRL B-4496 and L. acidophilus ATCC-4495, all other LAB strains produced PLA ranging from 0.021 to 0.275 mM. The high minimum inhibitory concentration for commercial PLA (3.01-36.10mM) suggests that it cannot be considered the only compound related with the antifungal potential of studied LAB and that synergistic effects may exist among other metabolism products.

Aspergillus niger time to growth in dried tomatoes.

Individual and combined effects of aw and incorporation of selected concentrations of Mexican oregano essential oil on the time to growth (TTG) of Aspergillus niger intentionally inoculated into dried tomatoes were studied during storage at 25°C for 100 days. For aw 0.96, 1,000 ppm of Mexican oregano essential oil inhibited A. niger growth during 100 days, whereas 500 ppm were sufficient at aw 0.91 and 250 ppm for tomatoes with aw 0.78. A. niger growth was evident at different incubation times depending on tested tomato aw and concentration of essential oil; these data were utilized to model TTG. Regression analysis revealed good agreement between experimental and predicted data with a correlation coefficient higher than 0.98. Analysis of mold growth data through TTG models makes possible to include observations detected as no growth and can be utilized to predict mold time to growth for specific preservation factor combinations or to select preservation factor levels for an expected shelf-life based on A. niger growth.

Thermal inactivation of Botrytis cinerea conidia in synthetic medium and strawberry puree.

Botrytis cinerea is one of the most important post-harvest molds that cause quality deterioration of strawberries and other fruits even during refrigeration storage. This research studied the effects of thermal inactivation of B. cinerea in synthetic medium and strawberry puree using hot water baths at different temperatures. These media were studied in order to determine if results obtained in a solution with the major components of the fruit (synthetic media), are comparable to the ones obtained in fruit purees. The results demonstrated that B. cinerea spores can be inactivated by heat treatments using relatively low temperatures (42-46 °C). Inactivation curves were well described by first order kinetics (R² 0.91-0.99). B. cinerea conidia inoculated in synthetic medium required less time to achieve one log reduction in population than those inoculated in the fruit puree. D values were 22, 8.5, 4 and 1.4 min at 42, 44, 46 and 48 °C, respectively, in synthetic medium; while D values in strawberry puree were 44.9, 13.8, 4.7 and 1.4 min at 42, 44, 46 and 48 °C, respectively. The z values obtained were 4.15 and 5.08 °C for the strawberry puree and synthetic medium respectively, showing higher sensitivity of B. cinerea in fruit purees than in the synthetic medium. Thus, a change in the medium composition had a marked difference in the heat inactivation of B. cinerea conidia, and the results obtained in synthetic medium are not accurate to describe the behavior of the microorganism in the fruit.

Zygosaccharomyces bailii inactivation by means of UV light and low-frequency ultrasound treatments.

Zygosaccharomyces bailii inactivation suspended in apple juice was evaluated under the effects of selected treatments: short-wave UV light (UVC, using one or two lamps), or low-frequency ultrasound (US), or their simultaneous combination. US treatments (20 kHz, 120-µm wave amplitude) were performed at 35°C in a double-wall vessel by using a 13-mm probe. The UVC device consists of two 90-cm-long stainless steel tubes with 40-W UVC lamps covered with quartz tubes, each one inside a stainless steel tube (annular inside diameter of 2.6 cm) connected to a peristaltic pump. Inoculated systems were recirculated through individual or simultaneous US and UVC treatments, samples were taken periodically, and yeast survivors were determined by the plate-count technique. Yeast survival curves demonstrated that UVC alone or in combination with US produced higher inactivation than US alone. Survival curves were appropriately described by the Weibull distribution of resistances model, obtaining model parameter values that adequately reflected the effect of the studied treatments. For every tested case, the distribution of resistances model revealed an absence of mode, while mean values and variances decreased when simultaneous UVC irradiation with two lamps and US were applied, reaching a 7-log cycle reduction after 40 min of treatment. Combined treatment was more effective than individual US or UVC treatments.

Colletotrichum gloeosporioides growth-no-growth interface after selected microwave treatments.

To study microwave heating for potential postharvest treatments against anthracnose disease, Colletotrichum gloeosporioides growth-no-growth response after selected microwave treatments (2,450 MHz) was fitted by using a logistic regression model. Evaluated variables were power level, exposure time, presence or absence of water in the medium during treatment, and incubation-observation time. Depending on the setting, the applied power ranged from 77.2 to 435.6 W. For the experiments on dry medium (mold spores over filter paper), exposure times were 1, 2, 3, or 4 min, whereas spores dispersed in potato dextrose agar, a wet medium, had exposure times of 3, 6, or 9 s. Growth (response = 1) or no growth (response = 0) was observed after two different incubation-observation times (4 or 10 days). As expected, high power levels and long exposure times resulted in complete inhibition of C. gloeosporioides spore germination. In a number of cases (such as low power levels and short treatment times), only a delay in mold growth was observed. Scanning electron micrographs showed signs of mycelia dehydration and structural collapse in the spores of the studied mold. Cell damage was attributed to heating during microwave exposure. Reduced logistic models included variables and interactions that significantly (P < 0.05) affected mold growth, and were able to predict the growth-no-growth response in at least 83% of the experimental conditions. Microwave treatments (4 min at any of the studied power levels in dry medium, and 9 s at power levels of 30% or more for wet medium) proved effective in the inhibition of C. gloeosporioides in model systems. These no-growth conditions will be tested further on fresh fruits in order to develop feasible postharvest microwave treatments.

Susceptibility of food-borne bacteria to binary combinations of antimicrobials at selected a(w) and pH.

AIM: To evaluate the antibacterial susceptibilities of food-borne bacteria to individual and binary mixtures of a synthetic antimicrobial agent with a natural phenolic compound. METHODS AND RESULTS: Antibacterial susceptibilities of Escherichia coli, Listeria innocua, Salmonella Typhimurium and Staphylococcus aureus to individual and binary mixtures of potassium sorbate with a phenolic compound (thymol, carvacrol, or eugenol) were evaluated, at selected water activity (a(w); 0.99 or 0.97) and pH (5.5 or 4.5). The bacteria studied were susceptible to the action of the antimicrobials individually with minimal inhibitory concentrations that varied from 800-ppm potassium sorbate for Staph. aureus at a(w) 0.99, and pH 5.5 to 100-ppm thymol or carvacrol for the four studied bacteria at a(w) 0.97 and pH 4.5. Several binary mixtures of potassium sorbate with thymol, carvacrol or eugenol inhibited bacterial growth. Antimicrobial agent inhibitory concentrations in the mixture varied among bacteria, additionally depending on the a(w) and the pH tested. CONCLUSIONS: Synergistic binary mixtures with fractional inhibitory concentration index <0.6 include 100- or 200-ppm potassium sorbate with 50- or 100-ppm thymol, carvacrol or eugenol. SIGNIFICANCE AND IMPACT OF THE STUDY: The synergistic combinations could be useful in reducing the amounts of antimicrobials needed to inhibit growth, thus diminishing consumer concerns regarding chemical preservatives.

Performance of mycological media in enumerating desiccated food spoilage yeasts: an interlaboratory study.

Dichloran 18% glycerol agar (DG18) was originally formulated to enumerate nonfastidious xerophilic moulds in foods containing rapidly growing Eurotium species. Some laboratories are now using DG18 as a general purpose medium for enumerating yeasts and moulds, although its performance in recovering yeasts from dry foods has not been evaluated. An interlaboratory study compared DG18 with dichloran rose bengal chloramphenicol agar (DRBC), plate count agar supplemented with chloramphenicol (PCAC), tryptone glucose yeast extract chloramphenicol agar (TGYC), acidified potato dextrose agar (APDA), and orange serum agar (OSA) for their suitability to enumerate 14 species of lyophilized yeasts. The coefficient of variation for among-laboratories repeatability within yeast was 1.39% and reproducibility of counts among laboratories was 7.1%. The order of performance of media for recovering yeasts was TGYC > PCAC = OSA > APDA > DRBC > DG 18. A second study was done to determine the combined effects of storage time and temperature on viability of yeasts and suitability of media for recovery. Higher viability was retained at -18 degrees C than at 5 degrees C or 25 degrees C for up to 42 weeks, although the difference in mean counts of yeasts stored at -18 degrees C and 25 degrees C was only 0.78 log10 cfu/ml of rehydrated suspension. TGYC was equal to PCAC and superior to the other four media in recovering yeasts stored at -18 degrees C, 5 degrees C, or 25 degrees C for up to 42 weeks. Results from both the interlaboratory study and the storage study support the use of TGYC for enumerating desiccated yeasts. DG18 is not recommended as a general purpose medium for recovering yeasts from a desiccated condition.

Probabilistic modeling of Saccharomyces cerevisiae inhibition under the effects of water activity, pH, and potassium sorbate concentration.

Probabilistic microbial modeling using logistic regression was used to predict the boundary between growth and no growth of Saccharomyces cerevisiae at selected incubation periods (50 and 350 h) in the presence of growth-controlling factors such as water activity (a(w); 0.97, 0.95, and 0.93), pH (6.0, 5.0, 4.0, and 3.0), and potassium sorbate (0, 50, 100, 200, 500, and 1,000 ppm). The proposed model predicts the probability of growth under a set of conditions and calculates critical values of a(w), pH, and potassium sorbate concentration needed to inhibit yeast growth for different probabilities. The reduction of pH increased the number of combinations of a(w) and potassium sorbate concentration with probabilities to inhibit yeast growth higher than 0.95. With a probability of growth of 0.05 and using the logistic models, the critical pH values were higher for 50 h of incubation than those required for 350 h. With lower a(w) values and increasing potassium sorbate concentration the critical pH values increased. Logistic regression is a useful tool to evaluate the effects of the combined factors on microbial growth.

Saccharomyces cerevisiae thermal inactivation kinetics combined with ultrasound.

Inactivation kinetics of Saccharomyces cerevisiae during thermal treatments at moderate temperatures (45.0, 47.5, 50.0, 52.5, or 55.0 degrees C) combined with application of 20 kHz of ultrasound were evaluated. S. cerevisiae inactivation under the combined effects of heat and ultrasound followed first-order reaction kinetics, with decimal reduction times (D) that varied from 22.3 to 0.8 min. D values in treatments that combined heat and ultrasound were significantly smaller (P < 0.05) than D values obtained for thermal treatments and were more noticeable at temperatures below 50 degrees C. The dependence of the D value on temperature had a significantly (P < 0.05) greater z value for combined treatments. Yeast heat inactivation kinetics revealed decreased thermal resistance caused by ultrasound.

Individual and combined effects of vanillin and potassium sorbate on Penicillium digitatum, Penicillium glabrum, and Penicillium italicum growth.

The individual and combined effects of potassium sorbate and vanillin concentrations on the growth of Penicillium digitatum, P. glabrum, and P. italicum in potato dextrose agar adjusted to water activity 0.98 and pH 3.5 were evaluated. Inhibitory concentrations of potassium sorbate varied from 150 ppm for P. digitatum to 700 ppm for P. glabrum, and for vanillin from 1,100 ppm for P. digitatum and P. italicum and 1,300 ppm for P. glabrum. Fractional inhibitory concentration (FIC) isobolograms show curves deviated to the left of the additive line. Calculated FIC index varied from 0.60 to 0.84. FIC index as well as FIC isobolograms show synergistic effects on mold inhibition when vanillin and potassium sorbate are applied in combination.

Oscillatory high hydrostatic pressure inactivation of Zygosaccharomyces bailii.

Zygosaccharomyces bailii inactivation was evaluated in oscillatory high hydrostatic pressure (HHP) treatments at sublethal pressures (207, 241, or 276 MPa) and compared with continuous HHP treatments in laboratory model systems with a water activity (aw) of 0.98 and pH 3.5. The yeast was inoculated into laboratory model systems and subjected to HHP in sterile bags. Two HHP treatments were conducted: continuous (holding times of 5, 10, 15, 20, 30, 60, or 90 min) and oscillatory (two, three, or four cycles with holding times of 5 min and two cycles with holding times of 10 min). Oscillatory pressure treatments increased the effectiveness of HHP processing. For equal holding times, Z. bailii counts decreased as the number of cycles increased. Holding times of 20 min in HHP oscillatory treatments at 276 MPa assured inactivation (< 10 CFU/ml) of Z. bailii initial inoculum. Oscillatory pressurization could be useful to decrease Z. bailii inactivation time.

High hydrostatic pressure come-up time and yeast viability.

The effects of the come-up time at selected pressures (50 to 689 MPa) on Saccharomyces cerevisiae and Zygosaccharomyces bailii viability were evaluated at 21 degrees C. For Z. bailii the effects of the water activity (a(w)) of the suspension media and the stage of the growth cycle were also investigated. Pressure come-up times exerted an important effect on the yeast survival fraction, decreasing counts as pressure increased. An increased sensitivity to pressure treatments was observed with yeast cells from the exponential growth phase. Lethality increased as a(w) of the suspension media increased. For an a(w) of 0.98 and cells from the stationary growth phase, pressure treatments at less than 200 MPa had no effect on Z. bailii viability; however, no survivors (< 10 CFU/ml) were observed in treatments applied only for the time needed to reach pressures greater than 517 MPa. Yeast survivor curves showed an excellent fit (r > 0.996) when described by a phenomenological model based on the Fermi equation, S(P) = 1/¿1 + exp[(P - Pc)/k]¿, where S(P) is the survival fraction, P is the pressure, Pc is a critical pressure corresponding to 50% survival, and k is a constant representing the steepness of the curve.

Effect of oscillatory high hydrostatic pressure treatments on Byssochlamys nivea ascospores suspended in fruit juice concentrates.

The effect of continuous (689 MPa with holding times of 5, 15 or 25 min) and oscillatory (one, three or five cycles at 689 MPa with holding times of 1 s) high hydrostatic pressure treatments on the viability of Byssochlamys nivea ascospores suspended in apple and cranberry juice concentrates adjusted by dilution to water activities (aw) of 0.98 and 0.94 was evaluated at 21 and 60 degrees C. Inactivation of the initial spore inocula was achieved after three or five cycles of oscillatory pressurization at 60 degrees C when the aw was 0.98 in both fruit juices. With aw 0.94, the initial inocula were reduced by less than 1 log-cycle after five pressure cycles. Inactivation was not observed within 25 min with continuous pressurization at 60 degrees C. In treatments at 21 degrees C, no effect on spore viability was observed with continuous or oscillatory treatments.