Adaptation of Salmonella spp. in juice stored under refrigerated and room temperature enhances acid resistance to simulated gastric fluid

The objective of this study was to evaluate the acid resistance of Salmonella spp. adapted in juices stored under refrigeration and room temperatures to simulated gastric fluid (SGF, pH 1.5). Five Salmonella serovars, Agona, Gaminara, Michigan, Montevideo, and Poona were used in this study. Apple, orange, and tomato juices inoculated with five serovars were stored at refrigeration (7 degrees C) and room temperature (20 degrees C) for 24 h for adaptation. Acid resistances of serovars adapted in juice were determined in SGF at 37 degrees C. All acid-adapted Salmonella serovars in juices displayed enhanced survival time compared to non-adapted controls. Among serovars, S. Poona adapted in apple at 20 degrees C and orange juices at 7 and 20 degrees C showed >2.0 log cfu/ml survivors, while the other serovars decreased to non-detectable level or <2.0 log cfu/ml for 100 s in SGF. Unlike apple and orange juices, all serovars adapted in tomato juice survived with >2.0 log cfu/ml for 100 s. For D-values, all Salmonella serovars adapted in apple and tomato juice enhanced their acid resistances compared to orange juices. S. Agona adapted in tomato juice at 7 degrees C and S. Poona in apple juice at 20 degrees C had the highest D-values with 82.9 and 82.5s, respectively. Results showed that the adaptation in juice increased acid resistance in SGF and varied by serovar, juice type, and adaptation temperature. Therefore, this study indicates that the introduction of Salmonella spp. to an acidic juice environment during processing can enhance their ability to survive in a human stomach, possibly increasing the risk of a Salmonella outbreak by juice.     

Effect of low-temperature, high-pressure treatment on the survival of Escherichia coli O157:H7 and Salmonella in unpasteurized fruit juices

The destructive effect of high pressure (615 MPa) combined with low temperature (15 degrees C) on various strains of Escherichia coli O157:H7 and various serovars of Salmonella in grapefruit, orange, apple, and carrot juices was investigated. The three-strain cocktail of E. coli O157:H7 (SEA13B88, ATCC 43895, and 932) was found to be most sensitive in grapefruit juice (8.34-log reduction) and least in apple juice (0.41-log reductions) when pressurized at 615 MPa for 2 min at 15 degrees C. Correspondingly, no injured survivor was detected in grapefruit and carrot juices under similar treatment conditions. No Salmonella spp. were detected in a 2-min pressure treatment (615 MPa, 15 degrees C) of grapefruit and orange fruit juices. Except for Enteritidis, all four serovars tested in the present study have viability loss of between 3.92- and 5.07-log reductions when pressurized in apple juice at 615 MPa for 2 min at 15 degrees C. No injured cells were recovered from grapefruit and orange juices, whereas the same treatment demonstrated reduction in numbers of Salmonella serovars Agona and Muenchen in apple juices and to a lesser extent with Typhimurium, Agona, and Muenchen in carrot juice. The present study demonstrated that low-temperature, high-pressure treatment has the potential to inactivate E. coli O157:H7 strains and different Salmonella spp. in different fruit juices.     

Thermal resistance parameters for Shiga toxin-producing Escherichia coli in apple juice

The purpose of the present study was to determine the heat resistance of six non-O157 Shiga toxin-producing Escherichia coli (STEC) serotypes in comparison to E. coli O157:H7 in single-strength apple juice without pulp. The thermal parameters for stationary-phase and acid-adapted cells of E. coli strains from serogroups O26, O45, O103, O111, O121, O145, and O157:H7 were determined by using an immersed coil apparatus. The most heat-sensitive serotype in the present study was O26. Stationary-phase cells for serotypes O145, O121, and O45 had the highest D(56°C)-value among the six non-O157 serotypes studied, although all were significantly lower (P < 0.05) than that of E. coli O157:H7. At 60°C E. coli O157:H7 and O103 demonstrated the highest D-values (1.37 ± 0.23 and 1.07 ± 0.03 min, respectively). The D(62°C) for the most heat-resistant strain belonging to the serotype O145 was similar (P > 0.05) to that for the most resistant O157:H7 strain (0.61 ± 0.17 and 0.60 ± 0.09 min, respectively). The heat resistance for stationary-phase cells was generally equal to or higher than that of acid-adapted counterparts. Although E. coli O157:H7 revealed D-values similar to or higher than the individual six non-O157 STEC serotypes in apple juice, the z-values for most non-O157 STEC tested strains were greater than those of E. coli O157:H7. When data were used to calculate heat resistance parameters at a temperature recommended in U.S. Food and Drug Administration guidance to industry, the D(71.1°C) for E. coli O157:H7 and non-O157 STEC serotypes were not significantly different (P > 0.05).     

Survival of antibiotic resistant and antibiotic sensitive strains of E. coli O157 and E. coli O26 in food matrices

Escherichia coli O157:H7 or E. coli O26, which were AS (antibiotic sensitive), AR (laboratory created antibiotic resistant mutants), or naturally MAR (multi-antibiotic resistant), were inoculated into laboratory media, yoghurt or orange juice and their growth/survival monitored during enrichment at 37 degrees C or storage at 4 degrees C. The strains were also inoculated into minced beef and their thermal inactivation (D-values) examined at 55 degrees C, with and without a prior heat shock at 48 degrees C. The growth kinetics (lag phases, growth rates) of the VTEC (verocytotoxigenic E. coli), incubated over 24 h at 37 degrees C in laboratory media, were similar regardless of the presence or absence of antibiotic resistance. In yoghurt and orange juice, E. coli O157:H7 MAR died off significantly faster (P<0.05) than any of other VTEC strains examined. E. coli O157:H7 MAR was also found to be significantly more heat sensitive (P<0.05) than the other VTEC strains tested. The reasons for the observed differences in survival of the different VTEC strains and the link between antibiotic resistance and survival in VTEC organisms are discussed.     

Escherichia coli O157:H7 populations in ruminants can be reduced by orange peel product feeding

Foodborne pathogenic bacteria such as Escherichia coli O157:H7 are threats to the safety of beef. Citrus peel and dried orange pulp are by-products from citrus juice production that have natural antimicrobial effects and are often incorporated into least-cost ration formulations for beef and dairy cattle. This study was designed to determine if orange peel and pulp affected E. coli O157:H7 populations in vivo. Sheep (n = 24) were fed a cracked corn grain-based diet that was supplemented with a 50-50 mixture of dried orange pellet and fresh orange peel to achieve a final concentration (dry matter basis, wt/wt) of 0, 5, or 10% pelleted orange peel (OP) for 10 days. Sheep were artificially inoculated with 10(10) CFU of E. coli O157:H7 by oral dosing. Fecal shedding of E. coli O157:H7 was measured daily for 5 days after inoculation, after which all animals were humanely euthanized. At 96 h postinoculation, E. coli O157:H7 shedding was reduced (P < 0.05) in sheep fed 10% OP. Populations of inoculated E. coli O157:H7 were reduced by OP treatment throughout the gastrointestinal tract; however, this reduction reached significant levels in the rumen (P < 0.05) of sheep fed 10% OP diets. Cecal and rectal populations of E. coli O157:H7 were reduced (P < 0.05) by inclusion of both 5 and 10% OP diets. Our results demonstrate that orange peel products can be used as a preharvest intervention strategy as part of an integrated pathogen reduction scheme.     

Survival of Escherichia coli O157:H7 during storage in pressure-treated orange juice.

The effect of a high-pressure treatment on the survival of a pressure-resistant strain of Escherichia coli O157:H7 (NCTC 12079) in orange juice during storage at 3 degrees C was investigated over the pH range of 3.4 to 5.0. The pH of shelf-stable orange juice was adjusted to 3.4, 3.6, 3.9, 4.5, and 5.0 and inoculated with 10(8) CFU ml(-1) of E. coli O157:H7. The orange juice was then pressure treated at 400 MPa for 1 min at 10 degrees C or was held at ambient pressure (as a control). Surviving E. coli O157:H7 cells were enumerated at 1-day intervals during a storage period of 25 days at 3 degrees C. Survival of E. coli O157:H7 during storage was dependent on the pH of the orange juice. The application of high pressure prior to storage significantly increased the susceptibility of E. coli O157:H7 to high acidity. For example, after pressure treatment, the time required for a 5-log decrease in cell numbers was reduced from 13 to 3 days at pH 3.4, from 16 to 6 days at pH 3.6, and from >25 to 8 days at pH 3.9. It is evident that the use of high-pressure processing of orange juice in order to increase the juice's shelf-life and to inactivate pathogens has the added advantage that it sensitizes E. coli O157:H7 to the high acid conditions found in orange juice, which results in the survival of significantly fewer E. coli O157:H7 during subsequent refrigerated storage.     

Antibacterial effect of monocaprylin on Escherichia coli O157:H7 in apple juice

The antibacterial effect of low concentrations of monocaprylin on Escherichia coli O157:H7 in apple juice was investigated. Apple juice alone (control) or containing 2.5 mM (0.055%) or 5 mM monocaprylin was inoculated with a five-strain mixture of E. coli O157:H7 at approximately 6.0 log CFU/ml. The juice samples were stored at 23 or 4 degrees C for 14 or 21 days, respectively, and the population of E. coli O157:H7 was determined on tryptic soy agar plates supplemented with 0.6% yeast extract. At both storage temperatures, the population of E. coli O157:H7 in monocaprylin-supplemented juice samples was significantly lower (P < 0.05) than that in the control samples. The concentration of monocaprylin and the storage temperature had a significant effect on the inactivation of E. coli O157:H7 in apple juice. Monocaprylin at 5 mM was significantly more effective than 2.5 mM monocaprylin for killing E. coli O157:H7 in apple juice. Inactivation of E. coli O157:H7 by monocaprylin was more pronounced in juice stored at 23 degrees C than in the refrigerated samples. Results of this study indicated that monocaprylin is effective for killing E. coli O157:H7 in apple juice, but detailed sensory studies are needed to determine the organoleptic properties of apple juice containing monocaprylin.     

Thermal resistance parameters for pathogens in white grape juice concentrate

The heat resistance of Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes that were in stationary phase, had been exposed to high osmotic pressure, or were acid adapted was evaluated in white grape juice concentrate (58 degrees Brix, pH 3.3). The most heat-resistant cells of all three pathogens were those exposed to high osmotic pressure or in stationary phase. Unlike in single-strength juices, in concentrate the acid-adapted cells for all three pathogens were less heat resistant than were cells in the other physiological states. E. coli O157:H7 had the highest heat resistance for all temperatures tested (e.g., D62 degrees C = 1.8 +/- 0.3 min, with a z-value of 9.9 +/- 0.6 degrees C). L. monocytogenes exposed to high osmotic pressure had the highest z-value (12.3 +/- 1.2 degrees C), although its D-values for all temperatures tested were lower (e.g., D62 degrees C = 0.93 +/- 0.1 min) than those for E. coli O157:H7. Salmonella was the most sensitive of the pathogens under all conditions. Based on the results obtained in this study, one example of a heat treatment that will inactivate 5 log units of all three pathogens in white grape juice concentrate was calculated as 1.5 min at 71.1 degrees C (z = 10.3 degrees C). Validation studies confirmed the predicted D71 degrees C for E. coli O157:H7 exposed to high osmotic pressure.     

Survival of Escherichia coli O157:H7 and Salmonella in apple cider and orange juice as affected by ozone and treatment temperature

Inactivation of Escherichia coli O157:H7 and Salmonella in apple cider and orange juice treated with ozone was evaluated. A five-strain mixture of E. coli O157:H7 or a five-serovar mixture of Salmonella was inoculated (7 log CFU/ml) into apple cider and orange juice. Ozone (0.9 g/h) was pumped into juices maintained at 4 degrees C, ambient temperature (approximately 20 degrees C), and 50 degrees C for up to 240 min, depending on organism, juice, and treatment temperature. Samples were withdrawn, diluted in 0.1% peptone water, and surface plated onto recovery media. Recovery of E. coli O157:H7 was compared on tryptic soy agar (TSA), sorbitol MacConkey agar, hemorrhagic coli agar, and modified eosin methylene blue agar; recovery of Salmonella was compared on TSA, bismuth sulfite agar, and xylose lysine tergitol 4 (XLT4) agar. After treatment at 50 degrees C, E. coli O157:H7 populations were undetectable (limit of 1.0 log CFU/ml; a minimum 6.0-log CFU/ml reduction) after 45 min in apple cider and 75 min in orange juice. At 50 degrees C, Salmonella was reduced by 4.8 log CFU/ml (apple cider) and was undetectable in orange juice after 15 min. E. coli O157:H7 at 4 degrees C was reduced by 4.8 log CFU/ml in apple cider and by 5.4 log CFU/ml in orange juice. Salmonella was reduced by 4.5 log CFU/ml (apple cider) and 4.2 log CFU/ml (orange juice) at 4 degrees C. Treatment at ambient temperature resulted in population reductions of less than 5.0 log CFU/ml. Recovery of E. coli O157:H7 and Salmonella on selective media was substantially lower than recovery on TSA, indicating development of sublethal injury. Ozone treatment of apple cider and orange juice at 4 degrees C or in combination with mild heating (50 degrees C) may provide an alternative to thermal pasteurization for reduction of E. coli O157:H7 and Salmonella in apple cider and orange juice.     

Destruction of Escherichia coli O157:H7 by vanillic acid in unpasteurized juice from six apple cultivars

The behavior of Escherichia coli O157:H7 in Granny Smith, Gala, Empire, McIntosh, Red Delicious, and Golden Delicious apple juice with or without supplementation with 5 or 10 mM vanillic acid was examined over a storage period of 7 days at 4 and 15 degrees C. The consequences of supplementation on sensory difference and preference were also determined by triangle testing. Juices made from the six apple cultivars had pH values ranging between pH 3.13 and 3.92. Vanillic acid exerted a concentration, pH, and time-dependent lethal effect toward E. coli O157:H7 in unpasteurized apple juice. Supplementation with 10 mM vanillic acid led to a 5-logarithm reduction in populations after 7 days at both temperatures, but sensory analysis revealed significant differences from and preference for unsupplemented juices. Supplementation with 5 mM vanillic acid accelerated death of E. coli O157:H7, but population reductions ranged from 5 log CFU/ml in low pH juices to none in high pH juices, particularly at 4 degrees C. No sensory difference or preference was detected in two of the six juices at this level of supplementation.     

Inactivation of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes in orange and tomato juice using ohmic heating

The effects of ohmic heating on reduction of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes in orange and tomato juice were investigated. Orange and tomato juice inoculated with E. coli O157:H7, Salmonella Typhimurium, and L. monocytogenes were subjected to ohmic heating with selected parameters including electric field strength from 10 to 20 V/cm and treatment times from 0 to 540 s. The number of pathogens was reduced by increasing the electric field strength from 10 to 20 V/cm as well as increasing treatment time. The population of E. coli O157:H7 was reduced more than 5 log after 120, 210, and 540 s of treatment in orange juice with 20, 15, and 10 V/cm electric field strengths, respectively. In tomato juice, levels of E. coli O157:H7 were reduced more than 5 log after 90, 180, and 480 s with the same electric field strengths. Similar phenomena were observed for Salmonella Typhimurium and L. monocytogenes, but E. coli O157:H7 was the most resistant to ohmic heating treatment. These results show that ohmic heating is potentially useful for inactivation of E. coli O157:H7, Salmonella Typhimurium, and L. monocytogenes and that the effect of inactivation depends on applied electric field strength, treatment time, pathogen species, and type of juice.     

Acid adaptation and temperature effect on the survival of E. coli O157:H7 in acidic fruit juice and lactic fermented milk product.

In this study, two strains of Escherichia coli O157:H7, (ATCC 43889 and ATCC 43895) were acid adapted at pH 5.0 in tryptic soy broth (TSB) for 4 h. Commercial products of mango juice (pH 3.2), asparagus juice (pH 3.6), Yakult--a diluted milk fermented drink (pH 3.6), and low-fat yoghurt (pH 3.9) were inoculated with acid-adapted or nonadapted cells of E. coli O157:H7. Survival of the inoculated E. coli O157:H7 in these commercial food products during storage at 25 or 7 degrees C was examined. It was found that although survival of the acid-adapted and nonadapted E. coli O157:H7 ATCC 43895 in asparagus juice during storage at 7 degrees C did not show marked difference, in general, acid adaptation and low temperature enhanced the survival of E. coli O157:H7 in both the commercial fruit juices tested. On the contrary, acid adaptation reduced the survival of both the strains of the test organism in Yakult and low-fat yoghurt stored at 7 degrees C. Besides, E. coli O157:H7 ATCC 43895 survived longer than ATCC 43889 in all the products examined, regardless of the storage temperature and acid adaptation.     

Response of Escherichia coli O157:H7 in apple and orange juices by hydrostatic pressure treatment with rapid decompression

The bactericidal efficiency of hydrostatic pressure treatment combined with a slow decompression (SD; about 30 s) or a rapid decompression (RD; about 2 ms) against clinically isolated Escherichia coli O157:H7 was investigated in apple juice, orange juice and McIlvaine buffers having the same pH values of the juices used. Effects of the SD and RD treatments on survivability of E. coli O157:H7 cells during storage at 4°C in the juices were also investigated. The RD treatment showed higher inactivation effect than the SD treatment in both the juices and buffers. Untreated E. coli O157:H7 cells were not inactivated during storage for 5 days; however, post-treatment storage after both the SD and RD treatments reduced survivability of E. coli O157:H7 cells in the juices. The degree of the reduction was higher in the cells subjected to the RD treatment than to the SD treatment.     

Fate of salmonellae in calcium-supplemented orange juice at refrigeration temperature.

Recent outbreaks of salmonellosis associated with orange juice have raised interest concerning the survival and growth of Salmonella in juice supplemented with calcium. A study was done to determine the influence of various calcium supplements on the survival of salmonellae in orange juice held at 4 degrees C for up to 32 days. Isolates of Salmonellal Muenchen (inoculum 1), Salmonella isolates from humans and animals (inoculum 2), and Salmonella isolates from produce outbreaks (inoculum 3) were inoculated into pasteurized orange juices with pH values ranging from 3.96 to 4.19 and containing 350 mg of calcium per 240-ml serving (1.46 mg of calcium/ml). Populations of Salmonella declined rapidly in juice containing calcium lactate (CaL), with counts decreasing from 4.86 log10 CFU/ml to < 1 log10 CFU/ml within 16 days, regardless of the Salmonella serotypes present in inoculum. Counts decreased from 4.89 log10 CFU/ml to < 1 log10 CFU/ml in orange juice supplemented with CaL and tricalcium phosphate (TCP) within 30 days. These reductions were significantly (P < or = 0.05) higher than those of the control (no calcium added), in which Salmonella populations decreased 3.19 +/- 0.20 log10 CFU/ml over 32 days. Populations in orange juice containing TCP or calcium citrate (CC) declined 1.34 +/- 0.20 log10 CFU/ml and 1.96 +/- 0.20 log10 CFU/ml, respectively, over 32 days. These counts were significantly higher than respective control counts in juice stored for 32 days. Populations of Salmonella of inoculum 3 inoculated into juice containing calcium citrate malate (CCM) were significantly higher than in the control. Higher numbers of cells in inoculum 3 also survived compared to numbers of cells of inocula 1 or 2 in juice supplemented with CCM. This study reveals that the form of calcium used to supplement orange juice influences the ability of salmonellae to survive.     

Acid tolerance and survival of Escherichia coli O157:H7 inoculated in fruit pulps stored under refrigeration.

The adaptation of Escherichia coli O157:H7 cells to acid conditions has been reported. This study showed the behavior of three strains of E. coli O157:H7 in two different physiological stages (acid shocked and control cells), inoculated in five fruit pulps stored at 4 degrees C for up to 30 days. The three strains of E. coli O157:H7 inoculated in grape pulp could be recovered up to day 30. E. coli O157:H7 strains survived for 4 days in all fruit pulps tested with different pH values (2.51 to 3.26), with the exception of acid-shocked cells of E. coli O157:H7 strain 933, which did not survive in "cajá" (Spodias lutea L.) pulp. The results clearly indicated that acid resistance can persist for long periods during storage at 4 degrees C. The protection conferred by acid adaptation suggests that acid-resistant organisms will be better equipped to outlast these acid challenges. The survival of E. coli O157:H7 in fruit pulps with a low pH, stored under refrigeration, is of extreme importance due to the high virulence of this microorganism. The viability of microorganisms was dependent on the viscosity index of fruit pulps. Less viscous pulps supported the bacteria survival longer than more viscous fruit pulps.     

Spread of bacterial pathogens during preparation of freshly squeezed orange juice

To study the potential of three bacterial pathogens to cross-contaminate orange juice during extraction, normal operation conditions during juice preparation at food service establishments were simulated. The spread of Salmonella enterica serovar Typhimurium, Escherichia coli O157:H7, and Listeria monocytogenes from inoculated oranges to work surfaces and to the final product was determined. The transference of these three bacterial pathogens to orange juice made from uninoculated oranges with the use of contaminated utensils was also studied. Fresh oranges were inoculated with a marker strain of rifampicin-resistant Salmonella Typhimurium, E. coli O157:H7, or L. monocytogenes. Final pathogen levels in juice were compared as a function of the use of electric or mechanical juice extractors to squeeze orange juice from inoculated oranges. Pathogen populations on different contact surfaces during orange juice extraction were determined on sulfite-phenol red-rifampicin plates for Salmonella Typhimurium and E. coli O157:H7 and on tryptic soy agar supplemented with 0.1 g of rifampicin per liter for L. monocytogenes. After inoculation, the average pathogen counts for the orange rind surface were 2.3 log10 CFU/cm2 for Salmonella Typhimurium, 3.6 log10 CFU/cm2 for E. coli O157:H7, and 4.4 log10 CFU/cm2 for L. monocytogenes. This contamination was spread over all utensils used in orange juice squeezing. Mean pathogen counts for the cutting board, the knife, and the extractor ranged from -0.3 to 2.1 log10 CFU/cm2, and the juice contained 1.0 log10 CFU of Salmonella Typhimurium per ml, 2.3 log10 CFU of E. coli O157:H7 per ml, and 2.7 log10 CFU of L. monocytogenes per ml. Contact with contaminated surfaces resulted in the presence of all pathogens in orange juice made from uninoculated oranges. These results give emphasis to the importance of fresh oranges as a source of pathogens in orange juice.     

Effect of vanillin on the fate of Listeria monocytogenes and Escherichia coli O157:H7 in a model apple juice medium and in apple juice

The effects of vanillin on the fates of Listeria monocytogenes and Escherichia coli O157:H7 at pH values between 3.5 and 4.5 were verified in a model apple juice (MAJ) medium and in apple juice incubated at 4 or 15 degrees C. Viable E. coli O157:H7 cells were recovered from MAJ for up to 10 days, but L. monocytogenes did not survive at pH 3.5. Supplementation with 40 mm vanillin exerted a lethal effect that was species, concentration, pH and temperature dependant. E. coli O157:H7 was more sensitive to vanillin than L. monocytogenes, and viable cells could not be recovered after 2 days incubation at either temperature. L. monocytogenes and E. coli O157:H7 were inoculated (10(5) cfu/ml) in pH adjusted (pH 4.00) or unadjusted (pH 3.42) juice from Granny Smith apples that was supplemented with 40 mm vanillin. Neither species were recovered after 3 days incubation at 4 or 15 degrees C. These findings indicate that vanillin could be useful as a preservative for minimally processed apple products.     

A Continuous High Pressure Carbon Dioxide System for Microbial Reduction in Orange Juice

ABSTRACT: A continuous high-pressure carbon dioxide system, run at ambient conditions, was tested on its performance in reducing both natural and inoculated microbial loads. The prototype system continuously processes orange juice with carbon dioxide (CO₂) at high pressures. A central composite design was originally used to examine the variables of pressure, residence time, and CO₂/juice ratio. For microbial reduction, residence time was the major factor followed by pressure. The CO₂/juice ratio showed no influence on microbial load, and in this equipment, was difficult to control. The unit was able to cause a 5-log reduction of the natural flora in spoiled juice, and could attain a 5-log decrease in numbers of pathogenic Escherichia coli O157:H7, Salmonella typhimurium , and Listeriamonocytogenes . No viable cells of E . coli O157:H7 orS. typhimurium were cultured after the treated juices were stored at room temperature (22 °C) for 14 d. Thus, non-thermal pathogen reduction is possible with this system.     

Evaluation of Escherichia coli O157:H7 in apple juice with Cornus fruit (Cornus officinalis Sieb. et Zucc.) extract by conventional media and thin agar layer method

Escherichia coli O157:H7 survival in apple juice supplemented with Cornus fruit (Cornus officinalis Sieb. et Zucc.) extract was studied. Inoculated samples with or without Cornus fruit extract were kept at 21 and 7 degrees C. Microbial analysis was conducted on days 0, 1, 3, 5, and 7. MacConkey sorbitol agar (MSA), tryptic soy agar (TSA), and thin agar layer (TAL) medium were used to compare the recovery of bacteria stressed under combination treatment. Influence of temperature, storage time, and Cornus fruit on survival of cells was evaluated. The most dramatic reduction of E. coli O157:H7 was observed in apple juice with Cornus fruit extract at 21 degrees C. At 7 degrees C, E. coli O157:H7 was reduced by 2.3logcfu/ml in the apple juice with Cornus fruit extract compared to the control sample on day 7. TAL and TSA were more efficient than MSA. Cornus fruit extract can be used in combination with temperature and storage time controls to inactivate E. coli O157:H7 in apple juice. This study has shown that TAL is a viable method of recovering and differentiating injured microorganisms and apple juice supplemented with Cornus fruit has potential as a value-added beverage with antimicrobial effects and potential health benefits.     

High-pressure resistance variation of Escherichia coli O157:H7 strains and Salmonella serovars in tryptic soy broth, distilled water, and fruit juice

The effect of high pressure on the log reduction of six strains of Escherichia coli O157:H7 and five serovars of Salmonella enterica was investigated in tryptic soy broth, sterile distilled water, and commercially sterile orange juice (for Salmonella) and apple cider (for E. coli). Samples were subjected to high-pressure processing treatment at 300 and 550 MPa for 2 min at 6 degrees C. Samples were plated onto tryptic soy agar directly after pressurization and after being held for 24 h at 4 degrees C. At 300 MPa, little effect was seen on E. coli O157:H7 strains, while Salmonella serovars varied in resistance, showing reductions between 0.26 and 3.95 log CFU/ml. At 550 MPa, E. coli O157:H7 strains exhibited a range of reductions (0.28 to 4.39 log CFU/ml), while most Salmonella populations decreased beyond the detection limit (> 5-log CFU/ml reduction). The most resistant strains tested were E. coli E009 and Salmonella Agona. Generally, bacterial populations in fruit juices showed larger decreases than did populations in tryptic soy broth and distilled water. E. coli O157:H7 cultures held for 24 h at 4 degrees C after treatment at 550 MPa showed a significant log decrease as compared with cultures directly after treatment (P < or = 0.05), while Salmonella serovars did not show this significant decrease (P > 0.05). All Salmonella serovars tested in orange juice treated at 550 MPa for 2 min at 6 degrees C and held for 24 h showed a > 5-log decrease, while E. coli O157:H7 strains require a higher pressure, higher temperature, longer pressurization, or a chemical additive to achieve a 5-log decrease.