Thermal inactivation of Salmonella and Escherichia coli O157:H7 on alfalfa seeds

Alfalfa seeds inoculated with five strains of Salmonella or Escherichia coli O157:H7 were subjected to dry heat at 55 degrees C for up to 8 days. Five-log reductions in Salmonella or E. coli O157:H7 on seeds were observed. No pathogens were detected on the sprouted seeds, which were initially inoculated with ca. 2 log CFU/g of Salmonella or more than 8 log CFU/g of E. coli O157:H7. The percentages of germination of the alfalfa seeds did not significantly decrease after 6 days of heating at 55 degrees C. These results showed that heat treatment of alfalfa seeds at 55 degrees C for up to 6 days was effective in enhancing the safety of alfalfa sprouts without affecting germination significantly.     

Cold plasma inactivates Salmonella Stanley and Escherichia coli O157:H7 inoculated on golden delicious apples

Cold plasma generated in a gliding arc was applied to outbreak strains of Escherichia coli O157:H7 and Salmonella Stanley on agar plates and inoculated onto the surfaces of Golden Delicious apples. This novel sanitizing technology inactivated both pathogens on agar plates, with higher flow rate (40 liters/min) observed to be more efficacious than were lower flow rates (20 liters/min), irrespective of treatment time (1 or 2 min). Golden Delicious apples were treated with various flow rates (10, 20, 30, or 40 liters/min) of cold plasma for various times (1, 2, or 3 min), applied to dried spot inoculations. All treatments resulted in significant (P < 0.05) reductions from the untreated control, with 40 liters/min more effective than were lower flow rates. Inactivation of Salmonella Stanley followed a time-dependent reduction for all flow rates. Reductions after 3 min ranged from 2.9 to 3.7 log CFU/ml, close to the limit of detection. For E. coli O157:H7, 40 liters/min gave similar reductions for all treatment times, 3.4 to 3.6 log CFU/ml. At lower flow rates, inactivation was related to exposure time, with 3 min resulting in reductions of 2.6 to 3 log CFU/ml. Temperature increase of the treated apples was related to exposure time for all flow rates. The maximum temperature of any plasma-treated apple was 50.8 degrees C (28 degrees C above ambient), after 20 liters/min for 3 min, indicating that antimicrobial effects were not the result of heat. These results indicate that cold plasma is a nonthermal process that can effectively reduce human pathogens inoculated onto fresh produce.     

Treatment of Escherichia coli O157:H7 inoculated alfalfa seeds and sprouts with electrolyzed oxidizing water

Electrolyzed oxidizing water is a relatively new concept that has been utilized in agriculture, livestock management, medical sterilization, and food sanitation. Electrolyzed oxidizing (EO) water generated by passing sodium chloride solution through an EO water generator was used to treat alfalfa seeds and sprouts inoculated with a five-strain cocktail of nalidixic acid resistant Escherichia coli O157:H7. EO water had a pH of 2.6, an oxidation-reduction potential of 1150 mV and about 50 ppm free chlorine. The percentage reduction in bacterial load was determined for reaction times of 2, 4, 8, 16, 32, and 64 min. Mechanical agitation was done while treating the seeds at different time intervals to increase the effectiveness of the treatment. Since E. coli O157:H7 was released due to soaking during treatment, the initial counts on seeds and sprouts were determined by soaking the contaminated seeds/sprouts in 0.1% peptone water for a period equivalent to treatment time. The samples were then pummeled in 0.1% peptone water and spread plated on tryptic soy agar with 5 microg/ml of nalidixic acid (TSAN). Results showed that there were reductions between 38.2% and 97.1% (0.22-1.56 log(10) CFU/g) in the bacterial load of treated seeds. The reductions for sprouts were between 91.1% and 99.8% (1.05-2.72 log(10) CFU/g). An increase in treatment time increased the percentage reduction of E. coli O157:H7. However, germination of the treated seeds reduced from 92% to 49% as amperage to make EO water and soaking time increased. EO water did not cause any visible damage to the sprouts.     

Inactivation of Escherichia coli O157:H7 on inoculated alfalfa seeds with ozonated water and heat treatment

Alfalfa seeds inoculated with a five-strain mixture of Escherichia coli O157:H7 were immersed in water containing 4, 8, 10, and 21 ppm of ozone for 2, 4, 8, 16, 32, and 64 min at 4 degrees C. Direct ozone sparging of seeds in water was used as an alternative mode of ozone treatment. Ozone-sparged seeds were also subsequently exposed to heat treatment at 40, 50, and 60 degrees C for 3 h. Populations of E. coli O157:H7 on untreated and treated seeds were determined by spread plating diluted samples on tryptic soy agar supplemented with 50 microg/ml of nalidixic acid. Since E. coli O157:H7 was released from inoculated seeds during treatment with ozone, the rate of release of cells from inoculated seeds soaked in 0.1% peptone water for up to 64 min was also determined. The overall reduction of E. coli O157:H7 on seeds treated with ozonated water without continuous sparging ranged from 0.40 to 1.75 log10 CFU/g (59.6 to 98.2%), whereas reductions for control seeds were 0.32 to 1.03 log10 CFU/g (51.7 to 90.5%). Treatment with higher ozone concentrations enhanced inactivation, but contact time longer than 8 min did not result in significantly higher reductions (P > 0.05). For seeds treated by ozone sparging, a 1.12-log10 CFU/g (92.1%) reduction was achieved using a 2-min contact time, and a 2.21-log10 CFU/g (99.4%) reduction was achieved with a 64-min contact time. The corresponding reductions for control seeds were 0.71 log10 CFU/g (79.5%) and 2.21 log10 CFU/g (99.4%), respectively. Treatment of ozone-sparged seeds at 60 degrees C for 3 h reduced the population to an undetectable level by direct plating (4 to 4.8 log10 CFU/g), although survivors were detected by enrichment. Ozone did not have a detrimental effect on seed germination percentage.     

Irradiation and chlorination effectively reduces Escherichia coli O157:H7 inoculated on cilantro (Coriandrum sativum) without negatively affecting quality

Cilantro (Coriandrum sativum) inoculated with Escherichia coli O157:H7 at levels approximating 10(7) CFU/g was dipped in 200 ppm chlorine solution followed by low-dose gamma irradiation. Samples were plated on tryptic soy agar containing 50 microg/ml nalidixic acid (TSAN) as well as TSAN plates with two 7-ml layers of basal yeast extract agar (TSAN-TAL). Levels of E. coli O157:H7 recovered from both types of media were determined over 11 days. Chlorination alone reduced counts by just over 1.0 log cycle, whereas irradiation at 1.05 kGy resulted in a 6.7-log reduction, and a combination of irradiation and chlorination reduced counts more than 7 log cycles. Trained panels performed analytical sensory tests at time intervals for 14 days to detect changes in yellowing, tip burn, browning, black rot, sliminess, off-aroma, and off-flavor. Sensory tests found no significant differences among attributes over time or dose in samples irradiated at 1.08 to 3.85 kGy. This study showed that combination treatments of chlorination and low-dose irradiation can significantly reduce levels of E. coli O157:H7 in fresh cilantro while maintaining product quality.     

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.     

Fate of Escherichia coli O157:H7 and Salmonella Enteritidis on currency.

The fate of foodborne pathogens Escherichia coli O157:H7 and Salmonella Enteritidis on coin surfaces was determined at room temperature (25 degrees C). A five-strain mixture of E. coli O157:H7 or Salmonella Enteritidis of approximately 5 x 10(4) CFU was applied to the surfaces of sterile U.S. coins (pennies, nickels, dimes, and quarters) and to the surfaces of two control substrata (Teflon and glass coverslips). During storage at room temperature, E. coli O157:H7 survived for 7, 9, and 11 days on the surfaces of pennies, nickels, and dimes and quarters, respectively. However, the pathogen died off within 4 to 7 days on both the Teflon and glass surfaces. Salmonella Enteritidis survived for 1, 2, 4, and 9 days on the surfaces of pennies, nickels, quarters, and dimes, respectively. Unlike E. coli O157:H7, survival of Salmonella Enteritidis was greatest on both Teflon and glass coverslips, with more than 100 cells per substratum detected at the 17th day of storage. Results indicate that coins could serve as potential vehicles for transmitting both E. coli O157:H7 and Salmonella Enteritidis.     

Irradiation to kill Escherichia coli O157:H7 and Salmonella on ready-to-eat radish and mung bean sprouts

A study was carried out to evaluate the effectiveness of ionizing radiation in eliminating Escherichia coli O157:H7 and Salmonella on commercial ready-to-eat radish and mung bean sprouts and to assess the chemical and physical quality of these sprouts. The use of ionizing radiation was investigated as a means of reducing or totally inactivating these pathogens, if present, on the sprouts. Treatment of mung bean and radish sprouts with a dose of 1.5 and 2.0 kGy, respectively, significantly reduced E. coli O157:H7 and Salmonella to nondetectable limits. The total vitamin C content was gradually reduced with the increase in irradiation dose (P < 0.0001). However, the effect of storage interval on the loss of vitamin C was nonsignificant for radish sprouts and significant for mung bean sprouts (P < 0.04). The color, firmness, and overall visual quality of the tested sprouts were acceptable when effective doses were applied to both radish and mung bean sprouts. Therefore, ionizing radiation could be useful in reducing the population of pathogens on sprouts and yet retain acceptable quality parameters.     

Inactivation kinetics of inoculated Escherichia coli O157:H7 and Salmonella enterica on lettuce by chlorine dioxide gas

The purpose of this investigation was to study inactivation kinetics of inoculated Escherichia coli O157:H7 and Salmonella enterica on lettuce leaves by ClO(2) gas at different concentrations (0.5, 1.0, 1.5, 3.0, and 5.0 mg l(-1)) for 10 min and to determine the effect of ClO(2) gas on the quality and shelf life of lettuce during storage at 4 degrees C for 7 days. One hundred microliters of each targeted organism was separately spot-inoculated onto the surface (5 cm(2)) of lettuce (approximately 8-9 log CFU ml(-1)), air-dried, and treated with ClO(2) gas at 22 degrees C and 90-95% relative humidity for 10 min. Surviving bacterial populations on lettuce were determined using a membrane transferring method, which included a non-selective medium followed by a selective medium. The inactivation kinetics of E. coli O157:H7 and S. enterica was determined using first-order kinetics to establish D-values and z-values. The D-values of E. coli and S. enterica were 2.9+/-0.1 and 3.8+/-0.5 min, respectively, at 5.0 mg l(-1) ClO(2) gas. The z-values of E. coli and S. enterica were 16.2+/-2.4 and 21.4+/-0.5 mg l(-1), respectively. A 5 log CFU reduction (recommended by the United States Food and Drug Administration) for E. coli and S. enterica could be achieved with 5.0 mg l(-1) ClO(2) gas for 14.5 and 19.0 min, respectively. Treatment with ClO(2) gas significantly reduced inherent microflora on lettuce and microbial counts remained significantly (p<0.05) lower than the uninoculated control during storage at 4 degrees C for 7 days. However, treatment with ClO(2) gas had a significantly (p<0.05) negative impact on visual leaf quality. These results showed that treatment with ClO(2) gas significantly reduced selected pathogens and inherent microorganisms on lettuce; however, the processing conditions would likely need to be altered for consumer acceptance.     

Inactivation of Escherichia coli O157:H7 and Salmonella spp. on alfalfa seeds by caprylic acid and monocaprylin

Alfalfa and other seed sprouts have been implicated in several Escherichia coli O157:H7 and Salmonella spp. human illness outbreaks in the U.S. Continuing food safety issues with alfalfa seeds necessitate the need for discovery and use of novel and effective antimicrobials. The potential use of caprylic acid (CA) and monocaprylin (MC) for reducing E. coli O157:H7 and Salmonella spp. populations on alfalfa seeds was evaluated. The effectiveness of three concentrations of CA and MC (25, 50, and 75 mM) to reduce E. coli O157:H7 and Salmonella spp. populations in 0.1% peptone water and on alfalfa seeds was evaluated. Surviving populations of E. coli O157:H7 and Salmonella spp. were enumerated by direct plating on tryptic soy agar (TSA). Non-inoculated alfalfa seeds were soaked for up to 120 min to evaluate the effect of CA and MC solutions on seed germination rate. For planktonic cells, the efficacy of the treatments was: 75 MC > 50 MC > 25 MC > 75 CA > 50 CA > 25 CA. Both E. coli O157:H7 and Salmonella spp. were reduced to below the detection limit (0.6 log CFU/ml) within 10 min of exposure to 75 MC from initial populations of 7.65 ± 0.10 log CFU/ml and 7.71 ± 0.11 log CFU/ml, respectively. Maximum reductions of 1.56 ± 0.25 and 2.56 ± 0.17 log CFU/g for E. coli O157:H7 and Salmonella spp., respectively, were achieved on inoculated alfalfa seeds (from initial populations of 4.74 ± 0.62 log CFU/g and 5.27 ± 0.20 log CFU/g, respectively) when treated with 75 MC for 90 min. Germination rates of CA or MC treated seeds ranged from 84% to 99%. The germination rates of CA or MC soaked seeds and water soaked seeds (control) were similar (P > 0.05) for soaking times of ≤ 90 min. Monocaprylin (75 mM) can be used to reduce E. coli O157:H7 and Salmonella spp. on alfalfa seeds without compromising seed viability.     

Survival of Salmonella and Escherichia coli O157:H7 in chorizos

Mexican-style raw meat sausages (chorizos) are not regulated in California when they are produced in small ethnic food markets. These sausages are sold uncooked, but their formulation imparts a color that may lead the consumer to assume that they are already cooked, and thus the chorizos may sometimes be eaten without proper cooking. If pathogens are present in such cases, illness may result. Survival of Salmonella and Escherichia coli O157:H7 in chorizos was evaluated under different storage conditions selected based on an initial survey of uninspected chorizos in California. Chorizos were formulated with five different initial water activity (aw) values (0.85, 0.90, 0.93, 0.95, and 0.97), stored under four conditions (refrigeration at 6 to 8 degrees C, room temperature at 24 to 26 degrees C, under a hood at 24 to 26 degrees C with forced air circulation, and incubation at 30 to 31 degrees C with convective air circulation), and sampled after 1, 2, 4, and 7 days. The initial pH was 4.8 and remained near 5.0 from day 1 of the sampling period. Two separate studies of packs inoculated with five-strain cocktails of Salmonella and of E. coli O157:H7 were performed twice for each initial aw. The three lowest aw values (0.85, 0.90, and 0.93) and the incubation and hood storage conditions were more effective (P < or = 0.05) at reducing the target pathogen levels in chorizos than were the two highest aw values (0.95 and 0.97) and the refrigeration storage condition, regardless of storage time. These results provide a scientific basis for guidelines given to producers of uninspected chorizo and should reduce the probability of foodborne illness associated with these products.     

Hot water treatments to inactivate Escherichia coli O157:H7 and Salmonella in mung bean seeds

The majority of the seed sprout-related outbreaks have been associated with Escherichia coli O157:H7 and Salmonella. Therefore, an effective method is needed to inactivate these organisms on the seeds before they are sprouted. This study was conducted to assess the effectiveness of various hot water treatments to inactivate E. coli O157:H7 and Salmonella populations on mung beans seeds intended for sprout production and to determine the effect of these treatments on seed germination after the seeds were dipped in chilled water for 30 s. Mung bean seed inoculated with four-strain cocktails of E. coli O157:H7 and Salmonella were soaked into hot water at 80 and 90 degrees C with shaking for various periods and then dipped in chilled water for 30 s. The treated seeds were then assessed for the efficacy of the treatment for reducing populations of the pathogens and the effects of the treatment on germination. After inoculation and air drying, 6.08 +/- 0.34 log CFU/g E. coli O157:H7 and 5.34 +/- 0.29 log CFU/g Salmonella were detected on the seeds. After hot water treatment at 90 degrees C for 90 s followed by dipping in chilled water for 30 s, no viable pathogens were found and no survivors were found in the enrichment medium and during the sprouting process. The germination yield of the seed was not affected significantly. Therefore, hot water treatment followed by dipping in chilled water for 30 s could be an effective seed decontamination method for mung bean seeds intended for sprout production.     

Reduction of Salmonella spp. and strains of Escherichia coli O157:H7 by gamma radiation of inoculated sprouts

There have been several recent outbreaks of salmonellosis and infections with Escherichia coli O157:H7 linked to the consumption of raw sprouts. Use of ionizing radiation was investigated as a means to reduce or to totally inactivate these pathogens, if present, on the sprouts. The radiation D value, which is the amount of irradiation in kilograys for a 1-log reduction in cell numbers, for these pathogens was established using a minimum of five doses at 19 +/- 1 degrees C. Before inoculation, the sprouts were irradiated to 6 kGy to remove the background microflora. The sprouts were inoculated either with Salmonella spp. cocktails made with either meat or vegetable isolates or with E. coli O157:H7 cocktails made with either meat or vegetable isolates. The radiation D values for the Salmonella spp. cocktails on sprouts were 0.54 and 0.46 kGy, respectively, for the meat and vegetable isolates. The radiation D values for the E. coli O157:H7 cocktails on sprouts were 0.34 and 0.30 kGy, respectively, for the meat and vegetable isolates. Salmonella was not detected by enrichment culture on sprouts grown from alfalfa seeds naturally contaminated with Salmonella after the sprouts were irradiated to a dose of 0.5 kGy or greater. Ionizing radiation is a process that can be used to reduce the population of pathogens on sprouts.     

Effect of plasma-activated organic acids against Salmonella Typhimurium and Escherichia coli O157:H7 inoculated on pork loin and its quality characteristics

This study investigated the effectiveness of plasma-activated organic acid (PAOA) on pork loin's bactericidal impact and quality. Salmonella Typhimurium, Escherichia coli O157:H7, and Listeria monocytogenes were used to inoculate the pork loins, revealing that PAOA achieved approximately 1.0 log CUF/g greater reduction than plasma-activated water. This was attributed to PAOA's lower pH, elevated oxidation-reduction potential, and increased reactive oxygen species concentrations. Notably, 0.5% plasma-activated acetic acid (PAA) showed 1.5 and 0.6 log CFU/g higher synergistic bactericidal effect against S. Typhimurium and E. coli O157:H7, respectively, compared to acetic acid. PAA also outperformed other PAOA treatments in terms of pork quality with similar meat color to deionized water, no adverse effect on lipid oxidation, and significantly reduced protein oxidation. Sensory analysis revealed PAA's highest umami taste. Thus, PAA is a promising method for microbial control in the meat industry while enhancing the oxidative stability and umami taste of pork loin.Industrial relevanceThis study demonstrated the industrial potential of combining plasma and organic acid treatments to control microorganisms in meat production and processing, while improving storage quality and sensory attributes. By using plasma- activated acetic acid, the oxidative stability of meat was enhanced during storage and umami taste was improved. This innovative technology offers advantages such as ease of operation and large-scale production, and it may help to overcome consumer concerns regarding the overuse of chlorine-based disinfectants, as organic acids are commonly used food additives.     

Reduction of Escherichia coli O157:H7 and Salmonella on laboratory-inoculated alfalfa seed with commercial citrus-related products

Alfalfa sprouts contaminated with the bacterial pathogens Escherichia coli O157:H7 and Salmonella have been the source of numerous outbreaks of foodborne illness in the United States and in other countries. The seed used for sprouting appears to be the primary source of these pathogens. The aim of this study was to determine whether the efficacy of commercial citrus-related products for sanitizing sprouting seed is similar to that of high levels of chlorine. Five products (Citrex, Pangermex, Citricidal, Citrobio, and Environné) were tested at concentrations of up to 20,000 ppm in sterile tap water and compared with buffered chlorine (at 16,000 ppm). Alfalfa seeds were inoculated with four-strain cocktails of Salmonella and E. coli O157:H7 to give final initial concentrations of ca. 9.0 and 7.0 CFU/g, respectively. Treatments (10 min) with Citrex, Pangermex, and Citricidal at 20,000 ppm and chlorine at 16,000 ppm produced similar log reductions for alfalfa seed inoculated with four-strain cocktails of E. coli O157:H7 and Salmonella (3.42 to 3.46 log CFU/g and 3.56 to 3.74 log CFU/g, respectively), and all four treatments were significantly (P<0.05) more effective than the control treatment (a buffer wash). Citrobio at 20,000 ppm was as effective as the other three products and chlorine against Salmonella but not against E. coli O157:H7. Environné was not more effective (producing reductions of 2.2 to 2.9 log CFU/g) than the control treatment (which produced reductions of 2.1 to 2.3 log CFU/g) against either pathogen. None of the treatments reduced seed germination. In vitro assays, as well as transmission electron microscopy, confirmed the antibacterial nature of the products that were effective against the two pathogens and indicated that they were bactericidal. When used at 20,000 ppm, the effective citrus-related products may be viable alternatives to chlorine for the sanitization of sprouting seed pending regulatory approval.     

Reduction of Escherichia coli O157:H7 and Salmonella spp. on laboratory-inoculated mung bean seed by chlorine treatmentt

Three U.S. outbreaks of foodborne illness due to consumption of contaminated raw mung bean sprouts occurred in the past 2 years and were caused by Salmonella Enteritidis. The original source of the pathogens is thought to have been the seed. The aim of this study was to determine whether treatment with aqueous chlorine would eliminate the pathogens from mung bean seed inoculated in the laboratory with four-strain cocktails of Escherichia coli O157:H7 and Salmonella spp. Treatments (for 5, 10, or 15 min) with buffered (500 mM potassium phosphate, pH 6.8) or unbuffered solutions containing 0.3 or 3.0% (wt/vol) Ca(OCl)2 were tested. In order to mimic common commercial practice, seed was rinsed before and after treatment with sterile tap water. Treatment for 15 min with buffer (500 mM potassium phosphate, pH 6.8) or sterile water in combination with the seed rinses resulted in maximum reductions of approximately 3 log10 CFU/g. The largest reductions (4 to 5 log10 CFU/g) for the chlorine treatments in combination with the rinses were obtained after treatment with buffered 3.0% (wt/vol) Ca(OCl)2 for 15 min. Treatment of mung bean seed for 15 min with unbuffered or buffered 3.0% (wt/vol) Ca(OCl)2 did not adversely affect germination. Even though treatments with 3% (wt/vol) Ca(OCl)2 in combination with the water rinses were effective in greatly reducing the populations of both bacterial pathogens, these treatments did not result in the elimination of the pathogens from laboratory-inoculated seed.     

Essential oils reduce Escherichia coli O157:H7 and Salmonella on spinach leaves

The efficacy of cinnamaldehyde and Sporan for reducing Escherichia coli O157:H7 and Salmonella on spinach leaves was investigated. Spinach leaves were inoculated with a five-strain cocktail of Salmonella or E. coli O157:H7, air dried for ca. 30 min, and then immersed in a treatment solution containing 5 ppm of free chlorine, cinnamaldehyde, or Sporan (800 and 1,000 ppm) alone or in combination with 200 ppm of acetic acid (20%) for 1 min or with water (control). After spin drying, treated leaves were analyzed periodically during 14 days of storage at 4°C for Salmonella, E. coli O157:H7, total coliforms, mesophilic and psychrotrophic bacteria, and yeasts and molds. Treatment effects on color and texture of leaves also were determined. Sporan alone (1,000S), Sporan plus acetic acid (1,000SV), and cinnamaldehyde-Tween (800T) reduced E. coli O157:H7 by more than 3 log CFU/g (P < 0.05), and 1,000SV treatment reduced Salmonella by 2.5 log CFU/g on day 0. E. coli O157:H7 and Salmonella populations on treated spinach leaves declined during storage at 4°C. The 1,000SV treatment was superior to chlorine and other treatments for reducing E. coli O157:H7 during storage. Saprophytic microbiota on spinach leaves increased during storage at 4°C but remained lower on leaves treated with Sporan (800S) and Sporan plus acetic acid (1,000SV) than on control leaves. The color and texture of Sporan-treated leaves were not significantly different from those of the control leaves after 14 days. Sporan plus acetic acid (1,000SV) reduced E. coli O157:H7 and Salmonella on baby spinach leaves without adverse effects on leaf color and texture.     

Response of Salmonella and Escherichia coli O157:H7 to UV energy

To determine the efficacy of a UV light treatment at 253.7 nm (UVC light) on microbial growth, plates containing tryptic soy agar plus 50 ppm of nalidixic acid (TSAN) were inoculated with known concentrations of five-strain cocktails of Salmonella and Escherichia coli O157:H7 and subjected to different UVC treatments. The concentration of the cocktail inoculum was determined with TSAN prior to inoculation. Serial dilutions were carried out, and inoculation levels of 10(0) to 10(8) CFU/ ml were tested for each pathogen. Multiple replications of doses of UV light ranging from 1.5 to 30 mW/cm2 were applied to different cocktail concentrations, and doses of > 8.4 mW/cm2 resulted in a 5-log reduction of Escherichia coli O157:H7, while a 5-log reduction of Salmonella was observed with doses of > 14.5 mW/cm2. Results for both organisms yielded sigmoidal inactivation curves. UVC light is effective in reducing microbial populations of pathogens on agar surfaces.     

Efficacy of chemical treatments in eliminating Salmonella and Escherichia coli O157:H7 on scarified and polished alfalfa seeds

Alfalfa seeds are sometimes subjected to a scarification treatment to enhance water uptake, which results in more rapid and uniform germination during sprout production. It has been hypothesized that this mechanical abrasion treatment diminishes the efficacy of chemical treatments used to kill or remove pathogenic bacteria from seeds. A study was done to compare the effectiveness of chlorine (20,000 ppm), H2O, (8%), Ca(OH)2 (1%), Ca(OH)2 (1%) plus Tween 80 (1%), and Ca(OH)2 (1%) plus Span 20 (1%) treatments in killing Salmonella and Escherichia coli O157:H7 inoculated onto control, scarified, and polished alfalfa seeds obtained from two suppliers. The influence of the presence of organic material in the inoculum carrier on the efficacy of sanitizers was investigated. Overall, treatment with 1% Ca(OH)2 was the most effective in reducing populations of the pathogens. Reduction in populations of pathogens on seeds obtained from supplier I indicate that chemical treatments are less efficacious in eliminating the pathogens on scarified seeds compared to control seeds. However, the effectiveness of chemical treatment in removing Salmonella and E. coli O157:H7 from seeds obtained from supplier 2 was not markedly affected by scarification or polishing. The presence of organic material in the inoculum carrier did not have a marked influence on the efficacy of chemicals in reducing populations of test pathogens. Additional lots of control, scarified, and polished alfalfa seeds of additional varieties need to be tested before conclusions can be drawn concerning the impact of mechanical abrasion on the efficacy of chemical treatment in removing or killing Salmonella and E. coli O157:H7.     

Chemical and irradiation treatments for killing Escherichia coli O157:H7 on alfalfa,radish, and mung bean seeds

In this study, the effectiveness of dry-heat treatment in combination with chemical treatments (electrolyzed oxidizing [EO] water, califresh-S, 200 ppm of active chlorinated water) with and without sonication in eliminating Escherichia coli O157:H7 on laboratory-inoculated alfalfa, radish, and mung bean seeds was compared with that of dry-heat treatment in combination with irradiation treatment. The treatment of mung bean seeds with EO water in combination with sonication followed by a rinse with sterile distilled water resulted in reductions of approximately 4.0 log10 CFU of E. coli O157:H7 per g. whereas reductions of ca. 1.52 and 2.64 log10 CFU/g were obtained for radish and alfalfa seeds. The maximum reduction (3.70 log10 CFU/g) for mung bean seeds was achieved by treatment with califresh-S and chlorinated water (200 ppm) in combination with sonication and a rinse. The combination of dry heat, hot EO water treatment, and sonication was able to eliminate pathogen populations on mung bean seeds but was unable to eliminate the pathogen on radish and alfalfa seeds. Other chemical treatments used were effective in greatly reducing pathogen populations on radish and alfalfa seeds without compromising the quality of the sprouts, but these treatments did not result in the elimination of pathogens from radish and alfalfa seeds. Moreover, a combination of dry-heat and irradiation treatments was effective in eliminating E. coli O157:H7 on laboratory-inoculated alfalfa, radish, and mung bean seeds. An irradiation dose of 2.0 kGy in combination with dry heat eliminated E. coli O157:H7 completely from alfalfa and mung bean seeds, whereas a 2.5-kGy dose of irradiation was required to eliminate the pathogen completely from radish seeds. Dry heat in combination with irradiation doses of up to 2.0 kGy did not unacceptably decrease the germination percentage for alfalfa seeds or the length of alfalfa sprouts but did decrease the lengths of radish and mung bean sprouts.