Inactivation of Escherichia coli O157:H7 on Inoculated Alfalfa Seeds with Ozonated Water and Heat Treatment

2002 ◽  
Vol 65 (3) ◽  
pp. 447-451 ◽  
Author(s):  
RATNA R. SHARMA ◽  
ALI DEMIRCI ◽  
LARRY R. BEUCHAT ◽  
WILLIAM F. FETT
Keyword(s):  
Escherichia Coli ◽  
Heat Treatment ◽  
Contact Time ◽  
Germination Percentage ◽  
Ozone Treatment ◽  
Escherichia Coli O157 ◽  
Alternative Mode ◽  
E Coli ◽  
Alfalfa Seeds ◽  
Ozonated Water

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°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°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 μg/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°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.

Chemical and Irradiation Treatments for Killing Escherichia coli O157:H7 on Alfalfa, Radish, and Mung Bean Seeds

2003 ◽  
Vol 66 (5) ◽  
pp. 767-774 ◽  
Author(s):  
M. L. BARI ◽  
E. NAZUKA ◽  
Y. SABINA ◽  
S. TODORIKI ◽  
K. ISSHIKI
Keyword(s):  
Escherichia Coli ◽  
Heat Treatment ◽  
Mung Bean ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Chemical Treatments ◽  
Dry Heat ◽  
Pathogen Populations ◽  
Dry Heat Treatment ◽  
Alfalfa 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.

Combination Treatments for Killing Escherichia coli O157:H7 on Alfalfa, Radish, Broccoli, and Mung Bean Seeds

2009 ◽  
Vol 72 (3) ◽  
pp. 631-636 ◽  
Author(s):  
M. L. BARI ◽  
D. NEI ◽  
K. ENOMOTO ◽  
S. TODORIKI ◽  
S. KAWAMOTO
Keyword(s):  
Escherichia Coli ◽  
Heat Treatment ◽  
Mung Bean ◽  
Germination Rate ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Dry Heat ◽  
Pathogen Populations ◽  
Dry Heat Treatment ◽  
Alfalfa Seeds

In this study, the effectiveness of prolonged dry-heat treatment (50°C) alone or in combination with chemical treatments (1% oxalic acid, 0.03% phytic acid, 50% ethanol, electrolyzed acidic water, and electrolyzed alkaline water) in eliminating Escherichia coli O157:H7 on laboratory-inoculated alfalfa, radish, broccoli, and mung bean seeds was compared with that of dry-heat treatment in combination with irradiation treatment. Dry-heat treatment for 17 or 24 h alone could reduce E. coli O157:H7 numbers to below detectable levels in radish, broccoli, and alfalfa seeds, but was unable to reduce the pathogen numbers to below the detectable level in mung bean seeds. In addition, dry-heat treatment for 17 h plus sanitizer treatments were effective in greatly reducing pathogen populations on radish, broccoli, and alfalfa seeds, without compromising the quality of the sprouts, but these treatments did not eliminate the pathogen from radish and alfalfa seeds. Seventeen hours of dry heat followed by a 1.0-kGy dose of irradiation completely eliminated E. coli O157:H7 from radish and mung bean seeds, whereas only a minimum radiation dose of 0.25 kGy was required to completely eliminate the pathogen from broccoli and alfalfa seeds. Dry heat in combination with radiation doses of up to 1.0 kGy did not negatively impact the seed germination rate or length of alfalfa, broccoli, and radish seeds or the length of alfalfa, broccoli, and radish sprouts, but did decrease the length of mung bean sprouts.

Comparison of Chemical Treatments to Eliminate Enterohemorrhagic Escherichia coli O157:H7 on Alfalfa Seeds

1999 ◽  
Vol 62 (4) ◽  
pp. 318-324 ◽  
Author(s):  
PETER J. TAORMINA ◽  
LARRY R. BEUCHAT
Keyword(s):  
Escherichia Coli ◽  
Germination Percentage ◽  
Enterohemorrhagic Escherichia Coli ◽  
Active Chlorine ◽  
Escherichia Coli O157 ◽  
Dramatic Decrease ◽  
E Coli ◽  
Surfactant Treatment ◽  
Chemical Treatments ◽  
Alfalfa Seeds

The focus of this study was to determine the efficacy of various chemicals in eliminating 2.04 to 3.23 log10 CFU/g of Escherichia coli O157:H7 from alfalfa seeds and to determine the survivability of the pathogen on seeds stored for prolonged periods at three temperatures. Significant (P ≤ 0.05) reductions in populations of E. coli O157:H7 on inoculated seeds were observed after treatments with 500 and 1,000 ppm of active chlorine (as Ca[OCl]2) for 3 but not 10 min and with ≥2,000 ppm of Ca(OCl)2 regardless of pretreatment with a surfactant. Treatment with 20,000 ppm of active chlorine failed to kill 2.68 log10 CFU/g of seeds. Acidified NaClO2 (500 ppm) was effective in reducing populations of the pathogen by >2 logs per g. Acidified ClO2 significantly reduced populations of E. coli O157:H7 on seeds at concentrations ≥100 ppm, and 500 ppm of ClO2 reduced the pathogen from 2.7 log10 CFU/g to <0.5 CFU/g. Chlorine (as NaOCl) was not effective at concentrations ≤1,000 ppm; significant reduction was achieved only after treatment with 2,000 ppm for 3 or 10 min. Notable reduction in populations was observed after treatment with 30 or 70% C2H3OH, but there was a dramatic decrease in germination percentage. Treatment with 0.2% H2O2 significantly reduced populations, and the organism was not detected by direct plating after treatment with ≥1% H2O2. Significant reduction in population of E. coli O157:H7 occurred after treatment with 1% trisodium phosphate, 40 ppm of Tsunami and Vortexx, and 1% Vegi-Clean. A significant decrease in the number of E. coli O157:H7 on dry seeds was observed within 1 week of storage at 25 and 37°C, but not at 5°C. Between 1 and 38 weeks, populations on seeds stored at 5°C remained relatively constant. The pathogen was recovered from alfalfa seeds initially containing 3.04 log10 CFU/g after storage at 25 or 37°C for 38 weeks but not 54 weeks.

Thermal Inactivation of Salmonella and Escherichia coli O157:H7 on Alfalfa Seeds

2007 ◽  
Vol 70 (7) ◽  
pp. 1698-1703 ◽  
Author(s):  
GUOPING FENG ◽  
JOHN J. CHUREY ◽  
RANDY W. WOROBO
Keyword(s):  
Escherichia Coli ◽  
Heat Treatment ◽  
Thermal Inactivation ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Dry Heat ◽  
Sprouted Seeds ◽  
Alfalfa Sprouts ◽  
Alfalfa Seeds

Alfalfa seeds inoculated with five strains of Salmonella or Escherichia coli O157:H7 were subjected to dry heat at 55°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°C. These results showed that heat treatment of alfalfa seeds at 55°C for up to 6 days was effective in enhancing the safety of alfalfa sprouts without affecting germination significantly.

Efficacy of Chemical Treatments in Eliminating Salmonella and Escherichia coli O157:H7 on Scarified and Polished Alfalfa Seeds

2001 ◽  
Vol 64 (10) ◽  
pp. 1489-1495 ◽  
Author(s):  
SARAH L. HOLLIDAY ◽  
ALAN J. SCOUTEN ◽  
LARRY R. BEUCHAT
Keyword(s):  
Escherichia Coli ◽  
Chemical Treatment ◽  
Organic Material ◽  
Pathogenic Bacteria ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Mechanical Abrasion ◽  
Chemical Treatments ◽  
The Impact ◽  
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), H2O2 (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 1 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.

Inactivation of Escherichia coli O157:H7, Salmonella Typhimurium DT104, and Listeria monocytogenes on Inoculated Alfalfa Seeds with a Fatty Acid–Based Sanitizer

2006 ◽  
Vol 69 (3) ◽  
pp. 582-590 ◽  
Author(s):  
PASCALE M. PIERRE ◽  
ELLIOT T. RYSER
Keyword(s):  
Escherichia Coli ◽  
Lactic Acid ◽  
Fatty Acid ◽  
Listeria Monocytogenes ◽  
Salmonella Typhimurium ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Reference Concentration ◽  
Glycerol Monolaurate ◽  
Alfalfa Seeds

Alfalfa seeds were inoculated with a three-strain cocktail of Escherichia coli O157:H7, Salmonella enterica subsp. enterica serovar Typhimurium DT104, or Listeria monocytogenes by immersion to contain ∼6 to 8 log CFU/g and then treated with a fatty acid–based sanitizer containing 250 ppm of peroxyacid, 1,000 ppm of caprylic and capric acids (Emery 658), 1,000 ppm of lactic acid, and 500 ppm of glycerol monolaurate at a reference concentration of 1×. Inoculated seeds were immersed at sanitizer concentrations of 5×, 10×, and 15× for 1, 3, 5, and 10 min and then assessed for pathogen survivors by direct plating. The lowest concentration that decreased all three pathogens by >5 log was 15×. After a 3-min exposure to the 15× concentration, populations of E. coli O157:H7, Salmonella Typhimurium DT104, and L. monocytogenes decreased by >5.45, >5.62, and >6.92 log, respectively, with no sublethal injury and no significant loss in seed germination rate or final sprout yield. The components of this 15× concentration (treatment A) were assessed independently and in various combinations to optimize antimicrobial activity. With inoculated seeds, treatment C (15,000 ppm of Emery 658, 15,000 ppm of lactic acid, and 7,500 ppm of glycerol monolaurate) decreased Salmonella Typhimurium, E. coli O157:H7, and L. monocytogenes by 6.23 and 5.57 log, 4.77 and 6.29 log, and 3.86 and 4.21 log after 3 and 5 min of exposure, respectively. Treatment D (15,000 ppm of Emery 658 and 15,000 ppm of lactic acid) reduced Salmonella Typhimurium by >6.90 log regardless of exposure time and E. coli O157:H7 and L. monocytogenes by 4.60 and >5.18 log and 3.55 and 3.14 log after 3 and 5 min, respectively. No significant differences (P > 0.05) were found between treatments A, C, and D. Overall, treatment D, which contained Emery 658 and lactic acid as active ingredients, reduced E. coli O157:H7, Salmonella Typhimurium, and L. monocytogenes populations by 3.55 to >6.90 log and may provide a viable alternative to the recommended 20,000 ppm of chlorine for sanitizing alfalfa seeds.

Effect of Heat Shock and Incubation Atmosphere on Injury and Recovery of Escherichia coli O157:H7

1993 ◽  
Vol 56 (7) ◽  
pp. 568-572 ◽  
Author(s):  
ELSA A. MURANO ◽  
MERLE D. PIERSON
Keyword(s):  
Escherichia Coli ◽  
Heat Treatment ◽  
Heat Shock ◽  
Escherichia Coli O157 ◽  
Toxic Substances ◽  
Anaerobic Conditions ◽  
E Coli ◽  
Heat Treated ◽  
Shock Response ◽  
Cell Components

Escherichia coli serotype O157:H7 cells were grown at 30°C for 6 h and subjected to a heat stress, or heat shock, at 42°C for 5 min. Heat-shocked and nonheat-shocked controls were heat treated at 55°C for up to 60 min. The number of injured cells was significantly higher in heat-shocked cells than in controls, and the rate of release of cell components was higher in heat-shocked cells. Anaerobic plating resulted in higher recovery of injured cells, when compared with aerobic plating, regardless of whether the cells were heat shocked or not. In addition, heat shocking resulted in lower catalase and superoxide dismutase activities when compared with controls. It also resulted in greater survivability after exposure to hydrogen peroxide, suggesting that heat shocking somehow enables the cells to survive exposure to toxic substances in addition to heat. The heat-shock response, coupled with anaerobic conditions, increased the ability of E. coli O157:H7 cells to recover after a heat treatment. Thus, heat shock did not afford protection to the cells against injury, but rather enhanced their ability to recover during storage.

Survival of Escherichia coli O157:H7 and Salmonella in Apple Cider and Orange Juice as Affected by Ozone and Treatment Temperature

2004 ◽  
Vol 67 (11) ◽  
pp. 2381-2386 ◽  
Author(s):  
ROBERT C. WILLIAMS ◽  
SUSAN S. SUMNER ◽  
DAVID A. GOLDEN
Keyword(s):  
Escherichia Coli ◽  
Ambient Temperature ◽  
Orange Juice ◽  
Treatment Temperature ◽  
Ozone Treatment ◽  
Escherichia Coli O157 ◽  
Apple Cider ◽  
E Coli ◽  
Peptone Water ◽  
Thermal Pasteurization

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°C, ambient temperature (approximately 20°C), and 50°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°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°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°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°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°C or in combination with mild heating (50°C) may provide an alternative to thermal pasteurization for reduction of E. coli O157:H7 and Salmonella in apple cider and orange juice.

Utilization of Gaseous Ozone for the Decontamination of Escherichia coli O157:H7 and Salmonella on Raspberries and Strawberries

2007 ◽  
Vol 70 (5) ◽  
pp. 1093-1098 ◽  
Author(s):  
KATHERINE L. BIALKA ◽  
ALI DEMIRCI
Keyword(s):  
Escherichia Coli ◽  
The United States ◽  
Ozone Treatment ◽  
Escherichia Coli O157 ◽  
Foodborne Illnesses ◽  
Treatment Scenario ◽  
Novel Technologies ◽  
E Coli ◽  
Gaseous Ozone ◽  
Ozone Maximum

Each year in the United States, there are approximately 76 million foodborne illnesses, and fresh produce is the second most common vehicle for such illnesses. Before going to market, small fruits are not washed or treated in any manner to extend their shelf life. Washing alone is not a viable option, and the use of novel technologies should be investigated. One such technology is ozone treatment, which has been used with drinking water since the late 19th century. The efficacy of gaseous ozone for killing pathogens on strawberries and raspberries, which were used as a model for small fruits, was investigated in this study. Strawberries and raspberries were artificially contaminated with five strains of Escherichia coli O157:H7 and Salmonella enterica. Fruits were treated with four ozone treatments: (i) continuous ozone flow (5%, wt/wt) for 2, 4, 8, 16, 32, and 64 min; (ii) pressurized ozone (83 kPa) for 2, 4, 8, 16, 32, and 64 min; (iii) continuous ozone (64 min) followed by pressurized ozone (64 min); and (iv) vacuum followed by 64 min of pressurized ozone. Maximum reductions for both strawberries and raspberries were achieved with the third treatment scenario. On strawberries, 2.60- and 2.96-log reductions were achieved for Salmonella and E. coli O157:H7, respectively. For raspberries, 3.55- and 3.75-log reductions were achieved for Salmonella and E. coli O157:H7, respectively. These results indicate that gaseous ozone should be a useful treatment for decontamination of small fruits.

Ozone Treatment for Reduction of Escherichia coli O157:H7 and Salmonella Serotype Typhimurium on Beef Carcass Surfaces

2003 ◽  
Vol 66 (5) ◽  
pp. 775-779 ◽  
Author(s):  
A. CASTILLO ◽  
K. S. MCKENZIE ◽  
L. M. LUCIA ◽  
G. R. ACUFF
Keyword(s):  
Escherichia Coli ◽  
Salmonella Typhimurium ◽  
Ozone Treatment ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Salmonella Serotype ◽  
Treatment Water ◽  
Beef Carcass

The effectiveness of an aqueous ozone treatment in reducing Escherichia coli O157:H7 and Salmonella serotype Typhimurium on hot carcass surfaces was determined with the use of a model carcass spray cabinet. Carcass surface regions were removed from carcasses and inoculated with feces containing 106 to 107 CFU each of E. coli O157:H7 and Salmonella Typhimurium per g and were then exposed to a water wash or to a water wash followed by a sanitizing ozone treatment. Water washes were applied at 28°C beginning at a pressure of 10 lb/in2 and gradually increasing to 400 lb/in2. Ozone treatment was carried out by spraying surfaces with an aqueous ozone solution (80 lb/in2 at 28°C) containing 95 mg of ozone per liter. Pathogen reductions achieved with ozone treatment were not significantly different from those achieved with a water wash alone. In addition, ozone treatment did not reduce E. coli O157:H7 or Salmonella Typhimurium contamination that was spread over the carcass surface as a result of the water wash. Under the conditions of this study, the aqueous ozone treatment applied resulted in no significant improvement over a water wash in reducing pathogens on beef carcass surfaces.

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