Glenn Boyd

Inactivation of Escherichia coli O157:H7 and Salmonella by Gamma Irradiation of Alfalfa Seed Intended for Production of Food Sprouts†

Inonizing irradiation was determined to be a suitable method for the inactivation of Salmonella and Escherichia coli O157:H7 on alfalfa seed to be used in the production of food sprouts. The radiation D (dose resulting in a 90% reduction of viable CFU) values for the inactivation of Salmonella and E. coli O157:H7 on alfalfa seeds were higher than the D-values for their inactivation on meat or poultry. The average D-value for the inactivation of Salmonella on alfalfa seeds was 0.97 +/- 0.03 kGy; the D-values for cocktails of meat isolates and for vegetable-associated isolates were not significantly different. The D-values for nonoutbreak and outbreak isolates of E. coli O157:H7 on alfalfa seeds were 0.55 +/- 0.01 and 0.60 +/- 0.01 kGy, respectively. It was determined that the relatively high D-values were not due to the low moisture content or the low water activity of the seed. The D-values for Salmonella on alfalfa seeds from two different sources did not differ significantly, even though there were significant differences in seed size and water activity. The increased moisture content of the seed after artificial inoculation did not significantly alter the D-value for the inactivation of Salmonella. The results of this study demonstrate that 3.3- and 2-log inactivations can be achieved with a 2-kGy dose of ionizing radiation, which will permit satisfactory commercial yields of sprouts from alfalfa seed contaminated with E. coli O157:H7 and Salmonella, respectively.

Effects of pH and acid resistance on the radiation resistance of enterohemorrhagic Escherichia coli

The effects of pH and the induction of pH-dependent stationary-phase acid resistance on the radiation resistance of Escherichia coli were determined for seven enterohemorrhagic strains and one nonenterohemorrhagic strain. The isolates were grown in acidogenic or nonacidogenic media to pH levels of approximately 4.7 and 7.2, respectively. The cells were then transferred to brain heart infusion (BHI) broth adjusted to pH 4.0, 4.5, 5.0, and 5.5 (with HCl) that was preequilibrated to 2 degrees C, and cultures were then irradiated using a 137Cs source. Surviving cells and the extent of injury were determined by plating on BHI and MacConkey agars both immediately after irradiation and after subsequent storage at 2 degrees C for 7 days. Decreasing the pH of the BHI in which E. coli was irradiated had relatively little effect on the microorganisms radiation resistance. Substantial differences in radiation resistance were noted among strains, and induction of acid resistance consistently increased radiation resistance. Comparison of E. coli levels immediately after irradiation and after 7 days of refrigerated storage suggested that irradiation enhanced pH-mediated inactivation of the pathogen. These results demonstrate that prior growth under conditions that induce a pH-dependent stationary phase cross-protects E. coli against radiation inactivation and must be taken into account when determining the microorganisms irradiation D value.

Radiation resistance ofSalmonella

SummaryThe ionizing radiation resistances of sixSalmonella species were examined. The experimental variables were the suspending medium, the presence or absence of air, and the temperature during the irradiation process.S. typhimurium ATCC 14028,S. enteritidis ATCC 9186,S. newport ATCC 6962,S. dublin ATCC 15480,S. anatum ATCC 9270, andS. arizonae ATCC 29933 were suspended in phosphate buffer (0.1 M, pH 7.0), brain heart infusion broth (BHI) or mechanically deboned chicken and exposed to gamma radiation from cesium-137 at 0.12 kGy per min. The radiation resistance of theSalmonella increased approximately two-fold when assayed in sterile mechanically deboned chicken rather than in buffer or BHI. The average radiation (0.30 to 1.20 kGy) D-value for all sixSalmonella strains was 0.56 kGy in mechanically deboned chicken.S. enteritidis was significantly more resistant to ionizing radiation than the other five strains ofSalmonella tested on mechanically deboned chicken. The temperature of irradiation but not the presence or absence of air significantly influenced the survival ofS. typhimurium andS. enteritidis in mechanically deboned chicken. Treatment of chicken meat with ionizing radiation would be an effective means for control ofSalmonella contamination.

Irradiation and modified atmosphere packaging for the control of Listeria monocytogenes on turkey meat.

When radiation-sterilized ground turkey meat was inoculated with Listeria monocytogenes, packaged under mixtures of nitrogen and carbon dioxide, and irradiated with gamma-radiation doses of 0 to 3.0 kGy, there was a statistically significant (P < 0.05), but probably not a biologically significant, lower (0.39 log) predicted bacterial survival in the presence of 100% carbon dioxide than in the presence of 100% nitrogen. Possibly because all atmospheres contained oxygen and because a response surface design was used, gamma-radiation resistance was not significantly (P < 0.05) different in air than in modified atmosphere packaging (MAP) mixtures containing 5% O2 or containing 20, 40, 60, and 80% CO2 and balance N2. The antilisterial effects of MAP mixtures containing 17.2, 40.5, and 64% CO2 and balance N2 were compared to those associated with air and vacuum packaging on turkey inoculated with approximately 5 x 10(3) CFU/g. Samples were irradiated to doses of 0, 0.5, 1.0, 1.5, 2.0, and 2.5 kGy and were stored at 7 degrees C for up to 28 days. Irradiation treatments were significantly more lethal in the presence of air packaging than in either vacuum packaging or MAP, and in those samples that received >1.0 kGy, there was a concentration-dependent CO2 inhibition of L. monocytogenes multiplication and/or recovery.

Effect of Irradiation Temperature on Inactivation of Escherichia coli O157:H7 and Staphylococcus aureus

The resistance of Escherichia coli O157:H7 and Staphylococcus aureus in ground beef to gamma radiation was significantly (P < 0.05) higher at subfreezing temperatures than above freezing. Ground beef was inoculated (ca. 2 x 10(8) CFU/g) with five isolates of either E. coli O157:H7 or S. aureus and subdivided into 25-g samples, vacuum packaged in barrier pouches, and tempered to 20, 12, 4, 0, -4, -12, -20, -30, -40, or -76 degrees C before gamma irradiation. The studies were repeated twice. The D10-values for both of these pathogens increased significantly at subfreezing temperatures, reaching maxima at approximately -20 degrees C. The D10-values for E. coli O157:H7 at 4 and -20 degrees C were 0.39 +/- 0.04 and 0.98 +/- 0.23 kGy, respectively. The D10-values for S. aureus at 0 and -20 degrees C were 0.51 degrees 0.02 and 0.88 +/- 0.05 kGy, respectively.

Atmospheric cold plasma inactivation of aerobic microorganisms on blueberries and effects on quality attributes

Cold plasma (CP) is a novel nonthermal technology, potentially useful in food processing settings. Berries were treated with atmospheric CP for 0, 15, 30, 45, 60, 90, or 120 s at a working distance of 7.5 cm with a mixture of 4 cubic feet/minute (cfm) of CP jet and 7 cfm of ambient air. Blueberries were sampled for total aerobic plate count (APC) and yeast/molds immediately after treatment and at 1, 2, and 7 days. Blueberries were also analyzed for compression firmness, surface color, and total anthocyanins immediately after each treatment. All treatments with CP significantly (P < 0.05) reduced APC after exposure, with reductions ranging from 0.8 to 1.6 log CFU/g and 1.5 to 2.0 log CFU/g compared to the control after 1 and 7 days, respectively. Treatments longer than 60s resulted in significant reductions in firmness, although it was demonstrated that collisions between the berries and the container contributed significantly to softening. A significant reduction in anthocyanins was observed after 90 s. The surface color measurements were significantly impacted after 120 s for the L* and a* values and 45 s for the b* values. CP can inactivate microorganisms on blueberries and could be optimized to improve the safety and quality of produce.

Radiation sensitization and postirradiation proliferation of Listeria monocytogenes on ready-to-eat deli meat in the presence of pectin-nisin films.

In this study, the ability of pectin-nisin films in combination with ionizing radiation to eliminate Listeria monocytogenes and inhibit its postirradiation proliferation was evaluated. Pectin films containing 0.025% nisin were made by extrusion. The surface of a ready-to-eat turkey meat sample was inoculated with L. monocytogenes at 10(6) CFU/cm2 and covered with a piece of pectin-nisin film. The samples were vacuum packaged and irradiated at 0, 1, and 2 kGy. The treated samples were stored at 10 degrees C and withdrawn at 0, 1, 2, 4, and 8 weeks for microbial analysis. Reductions in L. monocytogenes viability of 1.42, 1.56, 2.85, 3.78, and 5.36 log CFU/cm2 were achieved for the treatments of 1 kGy, pectin-nisin film, 2 kGy, 1 kGy plus pectin-nisin film, and 2 kGy plus pectin-nisin film, respectively. The greatest reduction (5.5 log CFU/cm2) was observed at 1 week for the 2 kGy plus pectin-nisin film treatment, suggesting that nisin was further released from the film to the surface of meat samples. Pectin-nisin films used in this study did not prevent but did significantly slow (P < 0.05) the proliferation of the L. monocytogenes cells that survived irradiation during 8 weeks of storage at 10 degrees C. These data indicate the potential use of pectin-nisin films alone or in combination with ionizing radiation for preventing listeriosis due to postprocessing contamination of ready-to-eat meat products.

Irradiation Inactivation of Four Salmonella Serotypes in Orange Juices with Various Turbidities

Reconstituted orange juice inoculated with Salmonella Anatum, Salmonella Infantis, Salmonella Newport, or Salmonella Stanley was treated with gamma radiation at 2 degrees C. To determine the relationship between juice antioxidant power and Dgamma (dose required to achieve 90% mortality), juice solids were removed prior to inoculation by centrifugation and/or filtration to create juice preparations of varying turbidity. In unadulterated orange juice, Salmonella Anatum (Dgamma = 0.71 kGy) was significantly more resistant than the other species tested. Salmonella Newport (Dgamma = 0.48 kGy) and Salmonella Infantis (Dgamma = 0.35 kGy) were significantly different, while Salmonella Stanley (Dgamma = 0.38 kGy) was intermediate between the two. Neither the resistance of each isolate nor the pattern of relative resistance among isolates was altered in reduced turbidity juice preparations. Although total antioxidant power was associated with the level of juice solids resuspended in phosphate buffer, antioxidant power was not significantly associated with turbidity in the juice preparations or with Dgamma of any species. The variable resistance to irradiation of the Salmonella isolates suggests this as a more significant factor than turbidity or antioxidant power in designing antimicrobial juice irradiation protocols.

Fate of gamma-irradiated Listeria monocytogenes during refrigerated storage on raw or cooked turkey breast meat.

The radiation resistance and ability of Listeria monocytogenes ATCC 7644, 15313, 43256, and 49594 to multiply on irradiated, air-packed, refrigerated raw or cooked turkey breast meat nuggets (ca. 25 g) and ground turkey breast meat was investigated. Gamma-radiation D values for L. monocytogenes were significantly different on raw and cooked nuggets, 0.56 +/- 0.03 kGy and 0.69 +/- 0.03 kGy, respectively; but they were not significantly different (P < or = 0.05) on raw and cooked ground turkey meat. High populations (approximately 10(9) CFU/g) of L. monocytogenes declined during 14 days of storage at 4 degrees C in both irradiated and nonirradiated samples of raw but not of cooked ground turkey breast meat. A moderate inoculum (approximately 10(3) CFU/g) did not survive a radiation dose of 3 kGy. The population increased in cooked but not in raw samples of irradiated ground turkey meat stored at either 2 or 7 degrees C for 21 days. The D value changed significantly from 0.70 +/- 0.04 to 0.60 +/- 0.02 kGy when the product was cooked to an internal temperature of 80 degrees C before irradiation. Growth on either raw or cooked turkey meat did not alter the radiation resistance of L. monocytogenes. Analyses were performed for pH, aw, moisture, and reducing potential of raw and cooked turkey meat and for pH, amino acid profile, thiamine, and riboflavin contents of aqueous extracts of raw and cooked turkey meats without identifying the factor or factors involved in differences in the survival and multiplication of L. monocytogenes on raw and cooked meat.

Reduction of Normal Flora by Irradiation and Its Effect on the Ability of Listeria monocytogenes to Multiply on Ground Turkey Stored at 7°C When Packaged under a Modified Atmosphere†

Listeria monocytogenes did not multiply faster during storage at 7 degrees C on irradiated than on nonirradiated raw ground turkey, and there was a concentration-dependent inhibition of its multiplication by CO2. Ground turkey was gamma irradiated at 5 degrees C to 0, 1.5, and 2.5 kGy and inoculated (approximately 100 CFU/g) after irradiation with a cocktail of L. monocytogenes ATCC 7644, 15313, 49594, and 43256. The meat was then packaged in air-permeable pouches or under atmospheres containing 30 or 53% CO2, 19% O2, and 51 or 24% N2 and stored at 7 degrees C for up to 28 days. A dose of 2.5 kGy extended the time for the total plate count (TPC) to reach 10(7) CFU/g from 4 to 19 days compared to that for nonirradiated turkey in air-permeable pouches. Following a dose of 2.5 kGy at the end of the 28-day study, the TPCs were 10(6.42) and 10(4.98) under 25% and 50% CO2 atmospheres, respectively. Under air, 30% CO2, and 53% CO2 atmospheres, the populations of L. monocytogenes after 19 days incubation were 10(4.89), 10(3.60), and 10(2.67) CFU/g. The populations of lactic acid bacteria and anaerobic or facultative bacteria were also reduced by irradiation. Irradiating ground turkey did not decrease its safety when it was contaminated following processing with L. monocytogenes.