Cold Atmospheric Plasma Jet Inactivates Cryptosporidium parvum Oocysts on Cilantro.

ABSTRACT: Cilantro was recently identified as a vehicle for protozoan illness. Current postharvest practices are not sufficient to inactivate protozoa on cilantro. Cold plasma is an emerging nonthermal waterless technology with potential applications in food processing that are currently being investigated to enhance the safety of herbs. The purpose of this study was to determine the impact of cold atmospheric plasma (CP) on the viability of Cryptosporidium parvum oocysts on cilantro. C. parvum oocysts were inoculated onto cilantro and treated with a CP jet for 0, 30, 90, and 180 s at a working distance of 10 cm with a flow of 1.42 × 10-3 m3/s. Oocyst viability was determined using HCT-8 cell culture infectivity assays. Overall, each treatment significantly reduced oocyst infectivity compared with the 0-s treatment control (P ≤ 0.02). Log inactivations of oocysts observed on cilantro were 0.84, 1.23, and 2.03 for the 30-, 90-, and 180-s treatment times, respectively. Drying and darkening of cilantro leaves was observed with treatments longer than 30 s. CP can reduce C. parvum infectivity on cilantro. With further research and optimization, this treatment technology has potential applications in postharvest processing of cilantro.

In-package atmospheric cold plasma treatment of bulk grape tomatoes for microbiological safety and preservation.

Effects of dielectric barrier discharge atmospheric cold plasma (DACP) treatment on the inactivation of Salmonella and the storability of grape tomato were investigated. Grape tomatoes, with or without inoculation with Salmonella, were packaged in a polyethylene terephthalate (PET) commercial clamshell container and cold plasma-treated at 35 kV at 1.1 A for 3 min using a DACP system equipped with a pin-type high-voltage electrode. DACP treatment inactivated Salmonella (p < 0.05) without altering the color or firmness of the grape tomatoes (p > 0.05). DACP treatment inactivated Salmonella uniformly in both layers of the double-layer configuration of the grape tomatoes regardless of the position of the tomatoes in each layer. Salmonella was most efficiently inactivated when the headspace to tomato volume ratio of the container was highest. Integration of rolling of tomatoes during treatment significantly increased the Salmonella reduction rates from 0.9 ±â€¯0.2 log CFU/tomato to 3.3 ±â€¯0.5 log CFU/tomato in the double-layer configuration of the tomato samples. Rolling-integrated DACP also initially reduced the number of total mesophilic aerobic bacteria and yeast and molds in the double-layer configuration of tomato samples by 1.3 ±â€¯0.3 and 1.5 ±â€¯0.2 log CFU/tomato, respectively. DACP treatment effectively reduced the growth of Salmonella and indigenous microorganisms at 10 and 25 °C, and did not influence the surface color, firmness, weight loss, lycopene concentration and residual ascorbic acid of grape tomatoes during storage at 10 and 25 °C. DACP treatment holds promise as a post-packaging process for improving microbial safety against Salmonella and storability of fresh grape tomatoes.

Cold Plasma Inactivation of Salmonella in Prepackaged, Mixed Salads Is Influenced by Cross-Contamination Sequence.

Customer demand for convenient food products has led to an increased production of prepackaged and ready-to-eat food products. Most of these products rely mainly on surface disinfection and other traditional approaches to ensure shelf life and safety. Novel processing techniques, such as cold plasma, are currently being investigated to enhance the safety and shelf life of prepacked foods. The purpose of this study was to determine the effects of cold plasma corona discharge on the inactivation of Salmonella on prepackaged, tomato-and-lettuce mixed salads. Two different inoculation methods were evaluated to address cross-contamination of Salmonella from cherry tomatoes to lettuce and vice versa. In separate studies, a sample of either cherry tomatoes (55 g) or romaine lettuce (10 g) was inoculated with a Salmonella cocktail (6.93 ± 0.99 log CFU/mL), placed into a commercial polyethylene terephthalate plastic container, and thoroughly mixed together with its noninoculated counterpart. Mixed salads were allowed to dry in a biosafety cabinet for 1 h. Samples were treated with 35 kV cold plasma corona discharge inside plastic containers for 3 min. Samples were stomached and serially diluted in buffered peptone water and then were plated onto aerobic plate count Petrifilm and incubated for 18 h at 37°C. When lettuce was the inoculated counterpart, log kill of Salmonella was significantly greater on tomatoes (0.75 log CFU/g) compared with lettuce (0.34 log CFU/g) (P = 0.0001). Salmonella was reduced on mixed salad only when lettuce was the inoculated counterpart (0.29 log CFU/g) (P = 0.002). Cold plasma can kill Salmonella in a prepackaged mixed salad, with efficacy dependent on the nature of contamination, direction of transfer, and the surface topography of the contaminated commodity.

In-package inhibition of E. coli O157:H7 on bulk Romaine lettuce using cold plasma.

Dielectric barrier discharge atmospheric cold plasma (DACP) treatment was evaluated for the inactivation of Escherichia coli O157:H7, surface morphology, color, carbon dioxide generation, and weight loss of bulk Romaine lettuce in a commercial plastic clamshell container. The lettuce samples were packed in a model bulk packaging configuration (three rows with either 1, 3, 5, or 7 layers) in the container and treated by DACP (42.6 kV, 10 min). DACP treatment reduced the number of E. coli O157:H7 in the leaf samples in the 1-, 3-, and 5-layer configurations by 0.4-0.8 log CFU/g lettuce, with no significant correlation to the sample location (P > 0.05). In the largest bulk stacking with 7 layers, a greater degree of reduction (1.1 log CFU/g lettuce) was observed at the top layer, but shaking the container increased the uniformity of the inhibition. DACP did not significantly change the surface morphology, color, respiration rate, or weight loss of the samples, nor did these properties differ significantly according to their location in the bulk stack. DACP treatment inhibited E. coli O157:H7 on bulk lettuce in clamshell containers in a uniform manner, without affecting the physical and biological properties and thus holds promise as a post-packaging process for fresh and fresh-cut fruits and vegetables.

Inactivation of Escherichia coli O157:H7 and Aerobic Microorganisms in Romaine Lettuce Packaged in a Commercial Polyethylene Terephthalate Container Using Atmospheric Cold Plasma.

The effects of dielectric barrier discharge atmospheric cold plasma (DACP) treatment on the inactivation of Escherichia coli O157:H7 and aerobic microorganisms in romaine lettuce packaged in a conventional commercial plastic container were evaluated during storage at 4°C for 7 days. Effects investigated included the color, carbon dioxide (CO2) generation, weight loss, and surface morphology of the lettuce during storage. Romaine lettuce pieces, with or without inoculation with a cocktail of three strains of E. coli O157:H7 (~6 log CFU/g of lettuce), were packaged in a polyethylene terephthalate commercial clamshell container and treated at 34.8 kV at 1.1 kHz for 5 min by using a DACP treatment system equipped with a pin-type high-voltage electrode. Romaine lettuce samples were analyzed for inactivation of E. coli O157:H7, total mesophilic aerobes, and yeasts and molds, color, CO2 generation, weight loss, and surface morphology during storage at 4°C for 7 days. The DACP treatment reduced the initial counts of E. coli O157:H7 and total aerobic microorganisms by ~1 log CFU/g, with negligible temperature change from 24.5 ± 1.4°C to 26.6 ± 1.7°C. The reductions in the numbers of E. coli O157:H7, total mesophilic aerobes, and yeasts and molds during storage were 0.8 to 1.5, 0.7 to 1.9, and 0.9 to 1.7 log CFU/g, respectively. DACP treatment, however, did not significantly affect the color, CO2 generation, weight, and surface morphology of lettuce during storage (P > 0.05). Some mesophilic aerobic bacteria were sublethally injured by DACP treatment. The results from this study demonstrate the potential of applying DACP as a postpackaging treatment to decontaminate lettuce contained in conventional plastic packages without altering color and leaf respiration during posttreatment cold storage.

Nonthermal inactivation of norovirus surrogates on blueberries using atmospheric cold plasma.

Viruses are currently the leading cause of foodborne outbreaks, most of which are associated with foods consumed raw. Cold plasma (CP) is an emerging novel nonthermal technology that can be used to surface decontaminate foods. This study investigated CP technology for the nonthermal inactivation of human norovirus surrogates, Tulane virus (TV) and murine norovirus (MNV), on the surface of blueberries. Blueberries (5 g) were weighed into sterile 4 oz. glass jars and inoculated with TV, 5 log PFU/g. Samples were treated with atmospheric CP for 0, 15, 30, 45, and 60 s at a working distance of 7.5 cm with 4 cubic feet/minute (cfm) of CP jet. Temperature readings were taken with an infrared camera prior to, and immediately following, CP treatments. In order to establish the impact of air flow during CP treatment (4 cfm), an additional 7 cfm jet of room temperature air was introduced from a separate nozzle. The experiment was repeated with 90 and 120 s as additional treatment time points. Viral titers were measured immediately after each treatment with a plaque assay using LLC-MK2 cells (TV) or RAW 264.7 cells (MNV). TV was significantly reduced 1.5 PFU/g compared to the control after treatment time of 45s, which was achieved regardless of temperature conditions. With the addition of 7 cfm of ambient air, the maximum log reduction for TV was 3.5 log PFU/g after 120s of treatment. MNV was significantly reduced by 0.5 log PFU/g compare to the control at 15s, and further treatment of MNV with ambient air brought the log reduction to greater than 5 log PFU/g at 90 s of treatment (Fig. 3). These results demonstrate that CP viral inactivation does not rely on thermal inactivation, and is therefore nonthermal in nature. With further optimization, CP may be used by food processors as a means of nonthermal inactivation of foodborne viruses.

Dielectric barrier discharge atmospheric cold plasma inhibits Escherichia coli O157:H7, Salmonella, Listeria monocytogenes, and Tulane virus in Romaine lettuce.

The present study investigated the effects of dielectric barrier discharge atmospheric cold plasma (DACP) treatment on the inactivation of Escherichia coli O157:H7, Salmonella, Listeria monocytogenes, and Tulane virus (TV) on Romaine lettuce, assessing the influences of moisture vaporization, modified atmospheric packaging (MAP), and post-treatment storage on the inactivation of these pathogens. Romaine lettuce was inoculated with E. coli O157:H7, Salmonella, L. monocytogenes (~6logCFU/g lettuce), or TV (~2logPFU/g lettuce) and packaged in either a Petri dish (diameter: 150mm, height: 15mm) or a Nylon/polyethylene pouch (152×254mm) with and without moisture vaporization. Additionally, a subset of pouch-packaged leaves was flushed with O2 at 5% or 10% (balance N2). All of the packaged lettuce samples were treated with DACP at 34.8kV for 5min and then analyzed either immediately or following post-treatment storage for 24h at 4°C to assess the inhibition of microorganisms. DACP treatment inhibited E. coli O157:H7, Salmonella, L. monocytogenes, and TV by 1.1±0.4, 0.4±0.3, 1.0±0.5logCFU/g, and 1.3±0.1logPFU/g, respectively, without environmental modifications of moisture or gas in the packages. The inhibition of the bacteria was not significantly affected by packaging type or moisture vaporization (p>0.05) but a reduced-oxygen MAP gas composition attenuated the inhibition rates of E. coli O157:H7 and TV. L. monocytogenes continued to decline by an additional 0.6logCFU/g in post-treatment cold storage for 24h. Additionally, both rigid and flexible conventional plastic packages appear to be suitable for the in-package decontamination of lettuce with DACP.

Reduction of Bacterial Pathogens and Potential Surrogates on the Surface of Almonds Using High-Intensity 405-Nanometer Light.

The disinfecting properties of high-intensity monochromatic blue light (MBL) were investigated against Escherichia coli O157:H7, Salmonella , and nonpathogenic bacteria inoculated onto the surface of almonds. MBL was generated from an array of narrow-band 405-nm light-emitting diodes. Almonds were inoculated with higher or lower levels (8 or 5 CFU/g) of pathogenic E. coli O157:H7 and Salmonella , as well as nonpathogenic E. coli K-12 and an avirulent strain of Salmonella Typhimurium, for evaluation as potential surrogates for their respective pathogens. Inoculated almonds were treated with MBL for 0, 1, 2, 4, 6, 8, and 10 min at a working distance of 7 cm. Simultaneous to treatment, cooling air was directed onto the almonds at a rate of 4 ft3/min (1.89 ×10-3 m3/s), sourced through a container of dry ice. An infrared camera was used to monitor the temperature readings after each run. For E. coli K-12, reductions of up to 1.85 or 1.63 log CFU/g were seen for higher and lower inoculum levels, respectively; reductions up to 2.44 and 1.44 log CFU/g were seen for E. coli O157:H7 (higher and lower inoculation levels, respectively). Attenuated Salmonella was reduced by up to 0.54 and 0.97 log CFU/g, whereas pathogenic Salmonella was reduced by up to 0.70 and 0.55 log CFU/g (higher and lower inoculation levels, respectively). Inoculation level did not significantly impact minimum effective treatment times, which ranged from 1 to 4 min. Temperatures remained below ambient throughout treatment, indicating that MBL is a nonthermal antimicrobial process. The nonpathogenic strains of E. coli and Salmonella each responded to MBL in a comparable manner to their pathogenic counterparts. These results suggest that these nonpathogenic strains may be useful in experiments with MBL in which a surrogate is required, and that MBL warrants further investigation as a potential antimicrobial treatment for low-moisture foods.

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.

Cold plasma rapid decontamination of food contact surfaces contaminated with Salmonella biofilms.

UNLABELLED: Cross-contamination of foods from persistent pathogen reservoirs is a known risk factor in processing environments. Industry requires a rapid, waterless, zero-contact, chemical-free method for removing pathogens from food contact surfaces. Cold plasma was tested for its ability to inactivate Salmonella biofilms. A 3-strain Salmonella culture was grown to form adherent biofilms for 24, 48, or 72 h on a test surface (glass slides). These were placed on a conveyor belt and passed at various line speeds to provide exposure times of 5, 10, or 15 s. The test plate was either 5 or 7.5 cm under a plasma jet emitter operating at 1 atm using filtered air as the feed gas. The frequency of high-voltage electricity was varied from 23 to 48 kHz. At the closer spacing (5 cm), cold plasma reduced Salmonella biofilms by up to 1.57 log CFU/mL (5 s), 1.82 log CFU/mL (10 s), and 2.13 log CFU/mL (15 s). Increasing the distance to 7.5 cm generally reduced the efficacy of the 15 s treatment, but had variable effects on the 5 and 10 s treatments. Variation of the high-voltage electricity had a greater effect on 10 and 15 s treatments, particularly at the 7.5 cm spacing. For each combination of time, distance, and frequency, Salmonella biofilms of 24, 48, and 72 h growth responded consistently with each other. The results show that short treatments with cold plasma yielded up to a 2.13 log reduction of a durable form of Salmonella contamination on a model food contact surface. This technology shows promise as a possible tool for rapid disinfection of materials associated with food processing. PRACTICAL APPLICATION: Pathogens such as Salmonella can form chemical-resistant biofilms, making them difficult to remove from food contact surfaces. A 15 s treatment with cold plasma reduced mature Salmonella biofilms by up to 2.13 log CFU/mL (99.3%). This contact-free, waterless method uses no chemical sanitizers. Cold plasma may therefore have a practical application for conveyor belts, equipment, and other food contact surfaces where a rapid, dry antimicrobial process is required.

Incorporation of preservatives in polylactic acid films for inactivating Escherichia coli O157:H7 and extending microbiological shelf life of strawberry puree.

Antimicrobial films of polylactic acid polymer incorporated with nisin, EDTA, sodium benzoate (SB), potassium sorbate (PS), and their combinations were developed, and their antimicrobial effects on the inactivation of Escherichia coli O157:H7 and natural background microflora (total aerobic bacteria, molds, and yeasts) in strawberry puree at 10 and 22 degrees C were determined. Direct addition of SB+PS to strawberry puree was also used as a comparison with SB+PS film treatment. The combination treatment reduced the cell populations of E. coli O157:H7 from 3.5 log CFU/ml to undetectable levels (<1 CFU/ml) after 14 days and 1 day at 10 and 22 degrees C, respectively, while the cells of E. coli O157:H7 in control samples survived up to 48 days at 10 degrees C and more than 14 days at 22 degrees C. The SB+PS film treatment produced a greater reduction of population of E. coli O157:H7 cells than did the SB+PS direct addition treatment. Similar results were observed for inactivation of natural microflora. In general, the antimicrobial effect was in the following order: film combination > SB+PS film > SB+PS direct addition > EDTA film > nisin film. The data obtained in this study suggest two approaches toward the development of control interventions against E. coli O157:H7 and extension of the microbiological shelf life of strawberry puree: (i) using antimicrobial packaging and (ii) using combinations of preservatives. The film formulas developed here can be used to make bottles or as coatings on the surface of bottles for use in liquid food packaging.

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.

Radiation sensitivity and postirradiation growth of foodborne pathogens on a ready-to-eat frankfurter on a roll product in the presence of modified atmosphere and antimicrobials.

Intervention technologies including ionizing radiation, antimicrobials, and modified atmospheres (MA) can be used to inhibit the growth of or inactivate foodborne pathogens on complex ready-to-eat foods such as sandwiches. However, the effect of these technologies when used in combination (the hurdle concept) on the survival of foodborne pathogens is unknown. The ability of ionizing radiation to inactivate Escherichia coli O157:H7, Salmonella, Listeria monocytogenes, and Staphylococcus aureus inoculated onto a frankfurter on a roll product containing the antimicrobials sodium diacetate and potassium lactate in the presence of an MA (100% N2, 50% N2 plus 50% CO2, or 100% CO2) was investigated. The radiation resistances (D10-values) of the foodborne pathogens were 0.43 to 0.47 kGy for E. coli O157:H7, 0.61 to 0.71 kGy for Salmonella, 0.53 to 0.57 for L. monocytogenes, and 0.56 to 0.60 for S. aureus. The MA had no effect on the radiation resistance of the pathogens. During a 2-week storage period under mild temperature abuse (10 degrees C), none of the pathogens were able to proliferate on the frankfurter on a roll product, regardless of the MA used. However, application of sublethal doses of ionizing radiation resulted in increased mortality of the gram-positive pathogens L. monocytogenes and S. aureus during the storage period regardless of the MA. Although the pathogens were unable to proliferate on the frankfurter on a roll product during the storage period, application of a postpackaging intervention step was needed to actually inactivate the foodborne pathogens. Ionizing radiation used in combination with sodium diacetate and potassium lactate resulted in additional mortality of L. monocytogenes and S. aureus, independent of the MA, during the 2-week storage period.

Elimination of Listeria monocytogenes from ready-to-eat turkey and cheese tortilla wraps using ionizing radiation.

Listeria monocytogenes is a common postprocess contaminant on ready-to-eat foods including premade ready-to-eat sandwiches. One popular type of sandwich product is the tortilla wrap, which contains sliced luncheon meats and cheeses rolled within a flour tortilla. This study determined the radiation resistance of L. monocytogenes surface inoculated onto two types of commercially available wheat flour tortillas, processed cheese slices, and deli turkey meat. The D10-values for L. monocytogenes (the radiation dose required to inactivate 1 log of the pathogen) were 0.27 kGy when inoculated onto two flour tortilla types, 0.28 and 0.30 kGy when inoculated onto two types of sliced processed cheeses, and 0.58 and 0.65 kGy when inoculated onto two types of sliced deli turkey meat. When two types of tortilla wraps were assembled from the individual components and L. monocytogenes was inoculated into the interfaces between the individual components, the D10-values were 0.27 to 0.37 kGy in the tortilla and cheese interfaces, 0.33 to 0.41 kGy in the cheese and turkey interfaces, and 0.25 to 0.33 kGy in the turkey and tortilla interfaces. The ability of ionizing radiation to reduce pathogen levels on the complex tortilla, cheese, and luncheon meat product was limited by the higher radiation resistance of L. monocytogenes when inoculated onto the ready-to-eat turkey-meat component.

Effect of freezing, irradiation, and frozen storage on survival of Salmonella in concentrated orange juice.

Six strains of Salmonella (Anatum F4317, Dublin 15480, Enteritidis 13076, Enteritidis WY15159, Stanley H0588, and Typhimurium 14028) were individually inoculated into orange juice concentrate (OJC) and frozen to -20 degrees C. The frozen samples were treated with 0 (nonirradiated), 0.5, 1.0, or 2.0 kGy of gamma radiation and held frozen for 1 h, and the surviving bacterial population was assessed. The strains showed significant variability in their response to freezing and to freezing in combination with irradiation. The response was dose dependent. Relative to the nonfrozen, nonirradiated control, the reduction following the highest dose (2.0 kGy) ranged from 1.29 log CFU/ml (Salmonella Typhimurium) to 2.17 log CFU/ml (Salmonella Stanley). Samples of OJC inoculated with Salmonella Enteritidis WY15159 and irradiated were stored at -20 degrees C for 1, 2, 7, or 14 days, and the surviving population was determined. Relative to the nonfrozen, nonirradiated control, after 14 days, the population was reduced by 1.2 log CFU/ml in the nonirradiated samples and by 3.3 log CFU/ml following treatment with 2.0 kGy. The combination of frozen storage plus irradiation resulted in greater overall reductions than either process alone.

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.

Effect of temperature on the radiation resistance of virulent Yersinia enterocolitica.

Yersinia enterocolitica, a food-borne pathogen, can be eliminated from meat using ionizing radiation. Commercial facilities may irradiate meat at refrigeration or frozen temperature, or packed in dry ice if the facility does not have refrigeration capabilities. The effect of temperature on the radiation resistance of Y. enterocolitica that contained the 70 kb large virulence plasmid was determined. A mixture of four Y. enterocolitica strains was inoculated into ground pork, which was then vacuum-packed, equilibrated to refrigeration or sub-freezing temperatures, and irradiated to doses of 0.2, 0.4, 0.6, 0.8, and 1.0 kGy. The D(10) value, the radiation dose required to reduce the number of viable Y. enterocolitica by 90%, increased as product temperature decreased with values of 0.19, 0.19, 0.21, 0.40, 0.40. 0.38, and 0.55 kGy being obtained at +5, 0, -5, -10. -15, -20 and -76 °C, respectively. Meat product temperature should be considered when selecting a radiation dose required for elimination of Y. enterocolitica.

Elimination by Gamma Irradiation of Salmonella spp. and Strains of Staphylococcus aureus Inoculated in Bison, Ostrich, Alligator, and Caiman Meat †.

There is an expanding industry for the marketing of high-value meats from animals other than the typical domesticated species, including, but not limited to, bison, ostrich, alligator, and caiman. In this study we compared the gamma radiation resistance of a mixture of salmonellae ( Salmonella dublin , S. enteritidis , S. newport , S. senftenberg , and S. typhimurium ) and a mixture of Staphylococcus aureus strains (ATCC 13565, ATCC 25923, and B124) when present on ground bison, ostrich, alligator, and caiman meats at 5°C. A minimum of five doses were used to establish the D values, and the studies were replicated three times. Because the type of meat did not significantly (P < 0.05) alter the radiation resistance of salmonellae and of S. aureus only slightly in the case of ostrich meat, all of the results for each organism were combined to obtain radiation D values of 0.53 ± 0.02 and 0.37 ± 0.01 kGy for Salmonella spp. and S. aureus , respectively. The authors conclude that both of these food-borne pathogens, if present, can be eliminated or greatly reduced in number, depending upon the level of contamination, from these meats by gamma radiation doses between 1.5 and 3.0 kGy at 5°C, the doses currently approved by the FDA and USDA for the irradiation of poultry. The authors also conclude that similar, if not identical, control of food-borne pathogens should be expected on edible meats in general, not just on those that are generically related.

Effects of NaCl, Sucrose, and Water Content on the Survival of Salmonella typhimurium on Irradiated Pork and Chicken.

We investigated the effects of water content, activity, sodium chloride (NaCl) and sucrose contents on the survival of Salmonella typhimurium ATCC 14028 in irradiated mechanically deboned chicken meat (MDCM) and ground pork loin. The effects of NaCl and sucrose concentration were investigated by adding various amounts to MDCM or ground pork loin, or by rehydrating freeze-dried ground pork loin with NaCl solutions with various degrees of saturation. The effects of water content were investigated by rehydrating freeze-dried ground pork loin with different quantities of water. Inoculated samples were irradiated at 5°C in vacuo to various doses up to 6.0 kGy. Highly significant effects (p < 0.01) of water content, water activity (aw) and NaCl content, but not of sucrose content, on the survival of S. typhimurium were identified. The failure of sucrose to provide the same protection for S. typhimurium in meat against radiation argues against reduced water activity being a primary mechanism of protection. The results indicate that the survival of foodborne pathogens on irradiated meats with reduced water content or increased NaCl levels may be greater than expected.

Effects of Ionizing Radiation and Anaerobic Refrigerated Storage on Indigenous Microflora, Salmonella , and Clostridium botulinum Types A and B in Vacuum-Canned, Mechanically Deboned Chicken Meat †.

Vacuum-canned, commercial, mechanically deboned chicken meat was challenged with either Clostridium botulinum spores (20 strains of types A and B, proteolytic; final spore concentration of ca. 400/g of meat) or Salmonella enteritidis (ca. 104 CFU/g of meat) followed by irradiation to 0, 1.5, and 3.0 kGy and storage at 5°C for 0, 2, and 4 weeks. None of the samples stored at 5°C developed botulinal toxin; however, when these samples were temperature abused at 28°C they became toxic within 18 h and had obvious signs of spoilage, i.e., swelling of the can and a putrid odor. During 4 weeks of refrigerated storage the log10 of the population of S. enteritidis in nonirradiated samples decreased from 3.86 to 2.58. S. enteritidis CFU were detectable in samples irradiated to 1.5 kGy at 0 weeks but not in samples irradiated to 3.0 kGy. Log levels of aerobic and facultative mesophiles increased during 4 weeks of refrigerated storage from 6.54 to 8.25, 4.03 to 8.14, and 2.84 to 5.23 in samples irradiated to 0, 1.5, and 3.0 kGy, respectively. Based on taxonomic analyses of 245 isolates, the bacterial populations depended upon radiation dose and storage time. The change was predominantly from gram-negative rods in nonirradiated samples to gram-positive streptococci in samples irradiated to 3.0 kGy and stored for 4 weeks. Spoilage organisms survived even the 3.0 kGy treatment.