Selection of surrogate bacteria in place of E. coli O157:H7 and Salmonella Typhimurium for pulsed electric field treatment of orange juice.

Pulsed electric field (PEF) technology has been used for the inactivation of microorganisms and to prevent flavor loss in liquid foods and beverages in place of thermal pasteurization. When used to pasteurize orange juice, PEF may prevent loss of volatile sensory attributes. Enterohemorrhagic E. coli O157:H7 (EHEC), two strains of Salmonella Typhimurium, and twenty strains of non-pathogenic bacteria were screened for inactivation in orange juice by PEF at 22 and 20kV/cm at 45 and 55 degrees C, respectively. Higher populations of both salmonellae were inactivated (2.81 and 3.54 log CFU/ml) at 55 degrees C, in comparison with the reduction of EHEC (2.22 log). When tested under the same conditions, inactivation of EHEC was slightly greater than that of a non-pathogenic E. coli (NPEC) ATCC 35218 (2.02 log). NPEC was further tested as a surrogate for EHEC by comparing inactivation kinetics at 45, 50 and 55 degrees C at field strengths of between 7.86 and 32.55kV/cm. Statistical comparison of revealed that EHEC and NPEC inactivation curves were homogeneous at outlet temperatures of 45 and 50 degrees C; however, EHEC was slightly more sensitive to PEF than the surrogate NPEC at 55 degrees C. The higher PEF resistance of non-pathogenic E. coli 35218 at 55 degrees C may provide a desirable margin of safety when used in pilot plant challenge studies in place of E. coli O157:H7.

Efficacy of supercritical carbon dioxide for nonthermal inactivation of Escherichia coli K12 in apple cider.

This study evaluated the efficacy of a supercritical carbon dioxide (SCCO(2)) system with a gas-liquid porous metal contactor for eliminating Escherichia coli K12 in apple cider. Pasteurized, preservative-free apple cider was inoculated with E. coli K12 and processed using the SCCO(2) system at CO(2) concentrations of 0-10% (wt.%, g CO(2)/100g product), outlet temperatures of 34, 38, and 42 degrees C, a system pressure of 7.6 MPa, and a flow rate of 1L/min. Increased CO(2) concentrations and temperatures significantly (P<0.05) enhanced the bactericidal effect, resulting in a maximum reduction of 7.31 log CFU/mL at 8% CO(2) and 42 degrees C. A response surface model indicated that minimum CO(2) concentrations of 9.9% at 34 degrees C, 7.4% at 38 degrees C, and 5.4% at 42 degrees C are needed to achieve a 5-log reduction of E. coli K12 in apple cider. SEM observations showed morphological changes in the cell envelope after SCCO(2) processing. At a processing condition of 8% and 38 degrees C, the reduction of E. coli was 6.03 log and the sublethal injury of the survivors was 84%. The regrowth or survival of E. coli in SCCO(2) processed apple cider was not observed during storage for 28 days at 4, 8, and 20 degrees C. Thus this study showed the potential of SCCO(2) processing with a gas-liquid porous metal contactor for the nonthermal pasteurization of apple cider.

Inactivation of Lactobacillus plantarum in apple cider, using radio frequency electric fields.

Radio frequency electric fields (RFEF) nonthermal processing effectively inactivates gram-negative bacteria in juices, but has yet to be shown effective at reducing gram-positive bacteria. Apple cider containing Lactobacillus plantarum ATCC 49445, a gram-positive bacterium, was RFEF processed under the following conditions: field strength of 0.15 to 15 kV/cm, temperature of 45 to 55 degrees C, frequency of 5 to 65 kHz, treatment time of 170 micros, and holding time of 5 to 50 s. The effect of refrigerating the inoculated cider prior to processing, the extent of sublethal injury, and the effect of storing the treated cider for 35 days were investigated. The population of L. plantarum was reduced by 1.0 log at 15 kV/cm, 20 kHz, and 50 degrees C, with a 5-s hold time. There is a synergistic effect between RFEF and heat above 50 degrees C. Inactivation significantly (P < 0.05) increased as frequency was decreased from 65 to 5 kHz. Inactivation increased linearly with field above 8 kV/cm. Holding cider at 55 degrees C after RFEF treatment for 5 and 50 s resulted in 2.5- and 3.1-log reductions, respectively. The surviving population was composed of 1.4-log sublethally injured cells. Storing processed cider at 4 degrees C for 35 days steadily and significantly (P < 0.05) reduced L. plantarum from 4.5 to 0.9 log CFU/ml. The electrical energy density was 51 J/ml. This provides the first evidence that nonthermal RFEF processing inactivates gram-positive bacteria, and that surviving cells may die off during refrigerated storage.

Impact of thermal and nonthermal processing technologies on unfermented apple cider aroma volatiles.

Aroma composition and microbial quality of identical lots of apple cider treated by pulsed electric field (PEF), ultraviolet irradiation (UV), or thermal pasteurization stored at 4 degrees C were compared at 0 and 4 weeks. Conditions were optimized to achieve identical 5 log reductions in Escherichia coli K12 for each treatment. PEF and thermal pasteurization maintained acceptable microbial quality for 4 weeks, but UV samples fermented after 2 weeks. Twenty-eight volatiles were quantified using gas chromatography-mass spectrometry (GC-MS) and odor activity values (OAV) determined. OAVs of 69:hexyl acetate, 41:hexanal, 25:2-methylbutyl acetate, 23:2-methyl ethyl butyrate, and 14:2-(E)-hexenal were observed for the control cider. Significant differences (p < 0.05) in the levels of these odorants were observed between treated apple ciders only after 4 weeks of storage. Thermal samples lost 30% of the major ester and aldehyde volatiles during storage with significant decreases (p < 0.05) in butyl acetate, 2-methylbutyl acetate, hexanal, and 2-(E)-hexenal. In UV cider, hexanal and 2-(E)-hexenal were completely lost after 4 weeks of storage. Microbial spoilage in UV cider after 4 weeks of storage was chemically confirmed by the detection of the microbial metabolite 1,3-pentadiene. PEF cider lost <2% of its total ester and aldehydes after 4 weeks of storage and was preferred by 91% of the sensory panel over thermally treated cider.

Antibrowning and antimicrobial properties of sodium acid sulfate in apple slices.

There are few available compounds that can both control browning and enhance microbial safety of fresh-cut fruits. In the present study, the antibrowning ability of sodium acid sulfate (SAS) on "Granny Smith" apple slices was first investigated in terms of optimum concentration and treatment time. In a separate experiment, the apple slices were treated with water or 3% of SAS, calcium ascorbate, citric acid, or acidified calcium sulfate for 5 min. Total plate count, color, firmness, and tissue damage were assessed during a 21-d storage at 4 degrees C. Results showed that the efficacy of SAS in inhibiting browning of apple slices increased with increasing concentration. A minimum 3% of SAS was needed to achieve 14 d of shelf life. Firmness was not significantly affected by SAS at 3% or lower concentrations. Antibrowning potential of SAS was similar for all treatment times ranging from 2 to 10 min. However, SAS caused some skin discoloration of apple slices. When cut surface of apple slices were stained with a fluorescein diacetate solution, tissue damage could be observed under a microscope even though visual damage was not evident. Among the antibrowning agents tested, SAS was the most effective in inhibiting browning and microbial growth for the first 14 d. Total plate count of samples treated with 3% SAS was significantly lower than those treated with calcium ascorbate, a commonly used antibrowning agent. Our results suggested that it is possible to use SAS to control browning while inhibiting the growth of microorganisms on the apple slices if the skin damage can be minimized. Practical Application: Fresh-cut apples have emerged as one of the popular products in restaurants, schools, and food service establishments as more consumers demand fresh, convenient, and nutritious foods. Processing of fresh-cut apples induces mechanical damage to the fruit and exposes apple tissue to air, resulting in the development of undesirable tissue browning. The fresh-cut industry currently uses antibrowning agents to prevent discoloration. However, the antibrowning solutions can become contaminated with human pathogens such as Listeria monocytogenes, and washing of apple slices with the contaminated solutions can result in the transfer of pathogens to the product. It would be ideal if an antibrowning compound prevented the proliferation of human pathogens in solutions and minimized the growth of pathogens during storage. The study was conducted to investigate antibrowning and antimicrobial properties of sodium acid sulfate (SAS) in comparison with other common antibrowning agents on Granny Smith apples. Results showed that among the antimicrobial agents we tested, SAS was the most effective in inhibiting browning and microbial growth for 14 d at 4 degrees C. However, SAS caused some skin discoloration of apple slices. Overall, SAS can potentially be used to inhibit tissue browning while reducing the microbial growth on apple slices. The information is useful for the fresh-cut produce industry to enhance microbial safety of fresh-cut apples while minimizing browning, thus increasing the consumption of the health benefiting fresh fruit.

Membrane damage and viability loss of Escherichia coli K-12 in apple juice treated with radio frequency electric field.

The need for a nonthermal intervention technology that can achieve microbial safety without altering nutritional quality of liquid foods led to the development of a radio frequency electric fields (RFEF) process. In order to understand the mechanism of inactivation of bacteria by RFEF, apple juice purchased from a wholesale distributor was inoculated with Escherichia coli K-12 at 7.8 log CFU/ml and then treated with RFEF. The inoculated apple juice was passed through an RFEF chamber operated at 20 kHz, 15 kV/cm for 170 micros at a flow rate of 540 ml/min. Treatment condition was periodically adjusted to achieve outlet temperatures of 40, 45, 50, 55, and 60 degrees C. Samples at each outlet temperature were plated (0.1 ml) and the number of CFU per milliliter determined on nonselective and selective agar media was used to calculate the viability loss. Bacterial inactivation and viability loss occurred at all temperatures tested with 55 degrees C treatment, leading to 4-log reductions. No significant effect was observed on bacterial population in control samples treated at 55 degrees C with a low-RFEF (0.15 kV/cm) field strength. These observations suggest that the 4-log reduction in samples treated at 15 kV/cm was entirely due to nonthermal effect. RFEF treatment resulted in membrane damage of the bacteria, leading to the efflux of intracellular ATP and UV-absorbing materials. Populations of injured bacteria recovered immediately (<30 min) from the treated apple juice averaged 0.43 log and were below detection after 1 h of RFEF treatment and determination using selective plates (tryptic soy agar containing 5% sodium chloride). The results of this study suggest that mechanism of inactivation of RFEF is by disruption of the bacterial surface structure leading to the damage and leakage of intracellular biological active compounds.

Comparison of effects of high-pressure processing and heat treatment on immunoactivity of bovine milk immunoglobulin G in enriched soymilk under equivalent microbial inactivation levels.

Immunoglobulin-rich foods may provide health benefits to consumers. To extend the refrigerated shelf life of functional foods enriched with bovine immunoglobulin G (IgG), nonthermal alternatives such as high-pressure processing (HPP) may offer advantages to thermal processing for microbial reduction. To evaluate the effects of HPP on the immunoactivity of bovine IgG, a soymilk product enriched with milk protein concentrates, derived from dairy cows that were hyperimmunized with 26 human pathogens, was subjected to HPP or heat treatment. To achieve a 5 log reduction in inoculated Escherichia coli 8739, the HPP or heat treatment requirements were 345 MPa for 4 min at 30 degrees C or for 20 s at 70 degrees C, respectively. To achieve a 5 log reduction in natural flora in the enriched soymilk, the HPP or heat treatments needed were 552 MPa for 4 min at 30 degrees C or for 120 s at 78.2 degrees C, respectively. At equivalent levels for a 5 log reduction in E. coli, HPP and heat treatment caused 25% and no detectable loss in bovine IgG activity, respectively. However, at equivalent levels for a 5 log reduction in natural flora, HPP and heat resulted in 65 and 85% loss of bovine IgG activity, respectively. Results of combined pressure-thermal kinetic studies of bovine milk IgG activity were provided to determine the optimal process conditions to preserve product function.