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

2016 ◽  
Vol 80 (1) ◽  
pp. 35-43 ◽  
Author(s):  
SEA C. MIN ◽  
SI HYEON ROH ◽  
GLENN BOYD ◽  
JOSEPH E. SITES ◽  
JOSEPH UKNALIS ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Weight Loss ◽  
Surface Morphology ◽  
Polyethylene Terephthalate ◽  
Cold Plasma ◽  
Escherichia Coli O157 ◽  
Romaine Lettuce ◽  
Plastic Container ◽  
E Coli ◽  
Yeasts And Molds

ABSTRACT 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.

Transfer of Escherichia coli O157:H7 to Romaine Lettuce due to Contact Water from Melting Ice

2008 ◽  
Vol 71 (2) ◽  
pp. 252-256 ◽  
Author(s):  
JIN KYUNG KIM ◽  
MARK A. HARRISON
Keyword(s):  
Escherichia Coli ◽  
Tap Water ◽  
Sampling Site ◽  
Contaminated Water ◽  
Escherichia Coli O157 ◽  
Romaine Lettuce ◽  
E Coli ◽  
Ice Melt ◽  
Leaf Surfaces ◽  
Shipping Containers

Ice can be used to chill romaine lettuce and maintain relative humidity during transportation. Escherichia coli O157:H7 may contaminate water used for ice. The objective of this study was to determine the potential for E. coli O157:H7 contamination of romaine lettuce from either ice contaminated with the pathogen or by transfer from lettuce surfaces via melting ice. In experiment 1, lettuce was spot inoculated with E. coli O157:H7 and chilled with ice prepared from uncontaminated tap water. In experiment 2, water inoculated with this pathogen was frozen and used to ice lettuce. Three heads of lettuce were stacked in each container and stored at 4 or 20°C. After the ice melted, E. coli O157:H7 attachment to and recovery from the lettuce leaves were determined. For experiment 1, the population of E. coli O157:H7 attached to inoculated sites averaged 3.8 and 5.5 CFU/cm2 at 4 and 20°C, respectively. Most of the uninoculated sites became contaminated with the pathogen due to ice melt. For experiment 2, 3.5 to 3.8 log CFU E. coli O157:H7 per cm2 was attached to the top leaf on the first head. After rinsing with chlorinated water (200 μg/ml), E. coli O157:H7 remained on the surface of the top head (1.8 to 2.0 log CFU/cm2). There was no difference in numbers of E. coli O157:H7 recovered from each sampling site at 4 and 20°C. Results show that E. coli O157:H7 can be transferred onto other produce layers in shipping containers from melted ice made of contaminated water and from contaminated to uncontaminated leaf surfaces.

Behavior of Escherichia coli O157:H7 on Damaged Leaves of Spinach, Lettuce, Cilantro, and Parsley Stored at Abusive Temperatures

2010 ◽  
Vol 73 (2) ◽  
pp. 212-220 ◽  
Author(s):  
ROWAIDA K. KHALIL ◽  
JOSEPH F. FRANK
Keyword(s):  
Escherichia Coli ◽  
Pathogenic Bacteria ◽  
Leaf Damage ◽  
Escherichia Coli O157 ◽  
Oxidation Reactions ◽  
Leaf Extracts ◽  
Romaine Lettuce ◽  
Growth And Survival ◽  
E Coli ◽  
Additional Information

Recent foodborne illness outbreaks associated with the consumption of leafy green produce indicates a need for additional information on the behavior of pathogenic bacteria on these products. Previous research indicates that pathogen growth and survival is enhanced by leaf damage. The objective of this study was to compare the behavior of Escherichia coli O157:H7 on damaged leaves of baby Romaine lettuce, spinach, cilantro, and parsley stored at three abusive temperatures (8, 12, and 15°C). The damaged portions of leaves were inoculated with approximately 105 CFU E. coli O157:H7 per leaf. The pathogen grew on damaged spinach leaves held for 3 days at 8 and 12°C (P < 0.05), with the population increasing by 1.18 and 2.08 log CFU per leaf, respectively. E. coli O157:H7 did not grow on damaged Romaine leaves at 8 or 12°C, but growth was observed after 8 h of storage at 15°C, with an increase of less than 1.0 log. Growth of E. coli O157:H7 on Romaine lettuce held at 8 or 12°C was enhanced when inocula were suspended in 0.05% ascorbic acid, indicating the possibility of inhibition by oxidation reactions associated with tissue damage. Damaged cilantro and Italian parsley leaves held at 8°C for 4 days did not support the growth of E. coli O157:H7. Behavior of the pathogen in leaf extracts differed from behavior on the damaged tissue. This study provides evidence that the damaged portion of a leafy green is a distinct growth niche that elicits different microbial responses in the various types of leafy greens.

Impact of Preinoculation Culture Conditions on the Behavior of Escherichia coli O157:H7 Inoculated onto Romaine Lettuce (Lactuca sativa) Plants and Cut Leaf Surfaces

2009 ◽  
Vol 72 (7) ◽  
pp. 1553-1559 ◽  
Author(s):  
CHRISTOPHER G. THEOFEL ◽  
LINDA J. HARRIS
Keyword(s):  
Escherichia Coli ◽  
High Standard ◽  
Culture Conditions ◽  
Escherichia Coli O157 ◽  
Late Stationary Phase ◽  
Romaine Lettuce ◽  
Preparation Methods ◽  
E Coli ◽  
Leaf Surfaces ◽  
Inoculum Preparation

Inoculum preparation methods can impact growth or survival of organisms inoculated into foods, thus complicating direct comparison of results among studies. The objective of this study was to evaluate preinoculation culture preparation for impact on Escherichia coli O157:H7 inoculated onto leaves of romaine lettuce plants and cut leaf surfaces. E. coli O157:H7 was grown quiescently or shaken at 15, 25, or 37°C to different growth phases in tryptic soy or M9 minimal salts broth or agar. Cells were harvested, washed, and suspended in 0.1% peptone, Milli Q water, or well water and refrigerated for 0 or 18 h. Prepared inoculum was spotted onto cut romaine lettuce (10 μl; 3 × 104 CFU/10 g) or onto romaine lettuce plants (20 μl; 3 × 106 CFU per leaf). Cut lettuce was sealed in 100-cm2 bags (made from a commercial polymer film) and incubated at 5 or 20°C. Lettuce plants were held at 23°C for 24 h. For all tested conditions, levels of E. coli O157:H7 increased at 20°Concut lettuce and decreased on cut lettuce stored at 5°C or on leaves of lettuce plants. At 20°C, preinoculation culture conditions had little impact on growth of E. coli O157:H7 on cut lettuce. However, survival at 5°C was significantly better (P < 0.05) for cultures grown at 15 or 37°C in minimal medium and to late stationary phase. Impact of preinoculation handling on survival on lettuce plants was less clear due to relatively high standard deviations observed among samples.

Comparative Examination of Escherichia coli O157:H7 Survival on Romaine Lettuce and in Soil at Two Independent Experimental Sites

2012 ◽  
Vol 75 (3) ◽  
pp. 480-487 ◽  
Author(s):  
GREG BEZANSON ◽  
PASCAL DELAQUIS ◽  
SUSAN BACH ◽  
ROBIN McKELLAR ◽  
ED TOPP ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Nalidixic Acid ◽  
Abiotic Factors ◽  
Decay Rates ◽  
Escherichia Coli O157 ◽  
Production Environment ◽  
Romaine Lettuce ◽  
Ecological Fitness ◽  
E Coli ◽  
Significant Difference

Little is known about the influence of abiotic factors such as climate and soil chemistry on the survival of Escherichia coli O157:H7 in field lettuce. We applied a nalidixic acid–resistant derivative of strain ATCC 700728 to field-grown romaine lettuce in two regions in Canada characterized by large variances in soil type and climate. Surviving populations in soil and on lettuce leaves were estimated on sorbitol MacConkey agar supplemented with nalidixic acid. Data were fitted with the Weibull decline function to permit comparison of decay rates in the two experimental sites. E. coli O157:H7 populations fell from 105 to <102 CFU/g on leaves, and <103 CFU/g in soil within 7 days after inoculation. Analysis revealed there was no significant difference between decay rates at the two experimental sites in either environment. The results of this study suggest that the inherent ecological fitness of E. coli O157:H7 ATCC 700728 determines the extent of survival in the production environment.

scholarly journals Outbreaks of Escherichia coli O157:H7 Infections Linked to Romaine Lettuce in Canada from 2008 to 2018: An Analysis of Food Safety Context

2020 ◽  
Vol 83 (8) ◽  
pp. 1444-1462 ◽  
Author(s):  
GENEVIÈVE COULOMBE ◽  
ANGELA CATFORD ◽  
AMALIA MARTINEZ-PEREZ ◽  
ENRICO BUENAVENTURA
Keyword(s):  
Escherichia Coli ◽  
The United States ◽  
Escherichia Coli O157 ◽  
Romaine Lettuce ◽  
E Coli ◽  
Leafy Greens ◽  
Iceberg Lettuce ◽  
Canadian Population ◽  
The Difference ◽  
Commercial Distribution

ABSTRACT Foodborne diseases are a major cause of illness in Canada. One of the main pathogens causing cases and outbreaks of foodborne illness in Canada is Escherichia coli O157:H7. From 2008 to 2018, 11 outbreaks of E. coli O157:H7 infection in Canada were linked to leafy greens, including 7 (63.6%) linked to romaine lettuce, 2 (18.2%) linked to iceberg lettuce, and 2 (18.2%) linked to other or unspecified types of leafy greens. The consumption of lettuce in Canada, the behavior of E. coli O157:H7 on lettuce leaves, and the production practices used for romaine and iceberg lettuce do not seem to explain why a higher number of outbreaks of E. coli O157:H7 infection were linked to romaine than to iceberg lettuce. However, the difference in the shape of iceberg and romaine lettuce heads could be an important factor. Among the seven outbreaks linked to romaine lettuce in Canada between 2008 and 2018, an eastern distribution of cases was observed. Cases from western provinces were reported only twice. The consumption of romaine and iceberg lettuce by the Canadian population does not seem to explain the eastern distribution of cases observed, but the commercial distribution, travel distances, and the storage practices used for lettuce may be important factors. In the past 10 years, the majority of the outbreaks of E. coli O157:H7 infection linked to romaine lettuce occurred during the spring (March to June) and fall (September to December). The timing of these outbreaks may be explained by the availability of lettuce in Canada, the growing region transition periods in the United States, and the seasonality in the prevalence of E. coli O157:H7. The consumption of romaine lettuce by the Canadian population does not explain the timing of the outbreaks observed. HIGHLIGHTS

Death of Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes in Shelf-Stable, Dairy-Based, Pourable Salad Dressings

2006 ◽  
Vol 69 (4) ◽  
pp. 801-814 ◽  
Author(s):  
LARRY R. BEUCHAT ◽  
JEE-HOON RYU ◽  
BARBARA B. ADLER ◽  
M. DAVID HARRISON
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Storage Period ◽  
Escherichia Coli O157 ◽  
Blue Cheese ◽  
Initial Inoculum ◽  
E Coli ◽  
Reduced Fat ◽  
Yeasts And Molds ◽  
Shelf Stable

The objectives of this study were to determine the death rates of Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes in three commercially manufactured full-fat ranch salad dressings, three reduced-fat ranch salad dressings, two full-fat blue cheese salad dressings, and two reduced-fat blue cheese salad dressings and to affirm the expectation that these dressings do not support the growth of these pathogens. The respective initial pH values of the four types of shelf-stable, dairy-based, pourable dressings were 2.87 to 3.72, 2.82 to 3.19, 3.08 to 3.87, and 2.83 to 3.49, respectively. Dressings were inoculated with low (2.4 to 2.5 log CFU/g) and high (5.3 to 5.9 log CFU/g) populations of separate five-strain mixtures of each pathogen and stored at 25°C for up to 15 days. Regardless of the initial inoculum population, all test pathogens rapidly died in all salad dressings. Salmonella was undetectable by enrichment (<1 CFU/25-ml sample in three replicate trials) in all salad dressings within 1 day, and E. coli O157:H7 and L. monocytogenes were reduced to undetectable levels by enrichment between 1 and 8 days and 2 and 8 days, respectively. E. coli O157:H7 was not detected in 4 of the 10 salad dressings stored for 2 or more days and 9 of the 10 dressings stored for 6 or more days after inoculation. L. monocytogenes was detected in 9 of the 10 salad dressings stored for 3 days but in only one dressing, by enrichment, at 6 days, indicating that it had the highest tolerance among the three pathogens to the acidic environment imposed by the dressings. Overall, the type of dressing (i.e., ranch versus blue cheese) and level of fat in the dressings did not have a marked effect on the rate of inactivation of pathogens. Total counts and populations of lactic acid bacteria and yeasts and molds remained low or undetectable (<1.0 log CFU/ml) throughout the 15-day storage period. Based on these observations, shelf-stable, dairy-based, pourable ranch and blue cheese salad dressings manufactured by three companies and stored at 25°C do not support the growth of Salmonella, E. coli O157:H7, and L. monocytogenes and should not be considered as potentially hazardous foods (time-temperature control for safety foods) as defined by the U.S. Food and Drug Administration Food Code.

Transfer of Escherichia coli O157:H7 from Simulated Wildlife Scat onto Romaine Lettuce during Foliar Irrigation

2015 ◽  
Vol 78 (2) ◽  
pp. 240-247 ◽  
Author(s):  
EDWARD R. ATWILL ◽  
JENNIFER A. CHASE ◽  
DAVID ORYANG ◽  
RONALD F. BOND ◽  
STEVEN T. KOIKE ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Bacterial Contamination ◽  
Assessment Model ◽  
Escherichia Coli O157 ◽  
Romaine Lettuce ◽  
E Coli ◽  
Weighted Arithmetic ◽  
Salinas Valley ◽  
Mean Concentration ◽  
Rabbit Feces

A field trial in Salinas Valley, California, was conducted during July 2011 to quantify the microbial load that transfers from wildlife feces onto nearby lettuce during foliar irrigation. Romaine lettuce was grown using standard commercial practices and irrigated using an impact sprinkler design. Five grams of rabbit feces was spiked with 1.29 × 108 CFU of Escherichia coli O157:H7 and placed −3, −2, and −1 days and immediately before a 2-h irrigation event. Immediately after irrigation, 168 heads of lettuce ranging from ca. 23 to 69 cm (from 9 to 27 in.) from the fecal deposits were collected, and the concentration of E. coli O157:H7 was determined. Thirty-eight percent of the collected lettuce heads had detectable E. coli O157:H7, ranging from 1 MPN to 2.30 × 105 MPN per head and a mean concentration of 7.37 × 103 MPN per head. Based on this weighted arithmetic mean concentration of 7.37 × 103 MPN of bacteria per positive head, only 0.00573% of the original 5 g of scat with its mean load of 1.29 × 108 CFU was transferred to the positive heads of lettuce. Bacterial contamination was limited to the outer leaves of lettuce. In addition, factors associated with the transfer of E. coli O157:H7 from scat to lettuce were distance between the scat and lettuce, age of scat before irrigation, and mean distance between scat and the irrigation sprinkler heads. This study quantified the transfer coefficient between scat and adjacent heads of lettuce as a function of irrigation. The data can be used to populate a quantitative produce risk assessment model for E. coli O157:H7 in romaine lettuce to inform risk management and food safety policies.

Modulation of Wound-Induced Hydrogen Peroxide and Its Influence on the Fate of Escherichia coli O157:H7 in Cut Lettuce Tissues

2012 ◽  
Vol 75 (12) ◽  
pp. 2208-2212 ◽  
Author(s):  
PETER M. A. TOIVONEN ◽  
CHANGWEN LU ◽  
SUSAN BACH ◽  
PASCAL DELAQUIS
Keyword(s):  
Escherichia Coli ◽  
Hydrogen Peroxide ◽  
Response To Treatment ◽  
Escherichia Coli O157 ◽  
Romaine Lettuce ◽  
Sodium Pyruvate ◽  
E Coli ◽  
Heat Treated ◽  
Cutting Heat ◽  
Microbial Analysis

Wounding of lettuce tissue has been examined previously by others in regard to browning reactions, and treatments to modulate wounding responses were evaluated for reduction of browning. However, the wounding process also releases oxygen radicals such as hydrogen peroxide. This study focused on the evaluation of two treatments that reduce hydrogen peroxide at cut surfaces (heat treatment and pyruvate addition) and one treatment that enhances its production (infusion with the fungal elicitor harpin). Hydrogen peroxide changes in response to treatment were also associated with resultant survival of Escherichia coli O157:H7, which was inoculated onto the lettuce before cutting. Heat-treated lettuce produced significantly less hydrogen peroxide, and microbial analysis showed that E. coli O157:H7 survival on packaged, heat-treated lettuce was higher than on non–heat-treated controls. Lettuce was also cut under a solution of sodium pyruvate (a well-known hydrogen peroxide quencher), and E. coli O157:H7 survival was found to be enhanced with that treatment. When lettuce was infused with harpin before cutting, hydrogen peroxide production was enhanced, and this was associated with reduced survival of E. coli O157:H7. These results collectively support the hypothesis that modulation of wound-generated hydrogen peroxide can have an influence on E. coli O157:H7 survival on cut and packaged romaine lettuce.

A Study of U.S. Orchards To Identify Potential Sources of Escherichia coli O157:H7†

2001 ◽  
Vol 64 (9) ◽  
pp. 1320-1327 ◽  
Author(s):  
D. C. R. RIORDAN ◽  
G. M. SAPERS ◽  
T. R. HANKINSON ◽  
M. MAGEE ◽  
A. M. MATTRAZZO ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Aerobic Bacteria ◽  
Escherichia Coli O157 ◽  
Total Coliforms ◽  
Apple Cider ◽  
E Coli ◽  
Fruit Samples ◽  
Potential Sources ◽  
Yeasts And Molds ◽  
The Individual

The association of unpasteurized apple cider with Escherichia coli O157:H7 foodborne illness has led to increased interest in potential reservoirs of this pathogen in the orchard. Fourteen U.S. orchards were surveyed in autumn 1999 to determine the incidence and prevalence of E. coli O157:H7, E. coli, total aerobic microflora, and yeasts and molds. Fruit samples (n = 63) (eight apple and two pear varieties) and soil, water, and fecal samples were collected. Samples were plated on (i) tryptic soy agar for total mesophilic aerobic count, (ii) E. coli and coliform Petrifilm for total coliforms and E. coli, and (iii) yeast and mold Petrifilm. Samples positive for coliforms and E. coli were enriched and tested for E. coli O157:H7. Fruit was also tested for internalization of microflora by aseptically removing the core, stem, and calyx areas, and the individual sections were assessed for the categories of microflora listed above. E. coli was detected in soil and water and in 6% of fruit samples (three pear samples and one apple sample), generally collected from areas previously designated as high risk in this study. However, no E. coli O157:H7 was found. Coliforms were found in 74% of fruit samples and were internalized in the cores of 40% of fruit tested. Yeasts and molds were internalized in 96.7% of samples and aerobic bacteria in 89.6%. E. coli was not found to be internalized. Total aerobic counts and total coliforms were higher in dropped and damaged fruit (P < 0.05). Findings suggest that dropped or damaged fruit should not be included in fruit designated for the production of unpasteurized juice or for the fresh or fresh-cut market. In addition, orchards should be located away from potential sources of contamination, such as pastures.

Occurrence of Coliform and Escherichia coli Contamination and Absence of Escherichia coli O157:H7 on Romaine Lettuce from Retail Stores in the Upper Midwest

2015 ◽  
Vol 78 (9) ◽  
pp. 1729-1732 ◽  
Author(s):  
JOSEPHINE D. GREVE ◽  
MARK S. ZIETLOW ◽  
KEVIN M. MILLER ◽  
JAY L. E. ELLINGSON
Keyword(s):  
Escherichia Coli ◽  
Microbial Contamination ◽  
Virulence Gene ◽  
Immunomagnetic Separation ◽  
Escherichia Coli O157 ◽  
Upper Midwest ◽  
Romaine Lettuce ◽  
E Coli ◽  
Recorded Data ◽  
Low Levels

A total of 720 whole, romaine lettuce heads were purchased from retail locations in the Upper Midwest and assessed for coliform and Escherichia coli contamination and for the presence of E. coli O157:H7. During a 16-month period (August 2010 through December 2011), coliform and E. coli counts were enumerated on Petrifilm, and the presence of E. coli O157:H7 and the virulence gene eae was evaluated by real-time PCR (qPCR). Over half (400 of 720) of the lettuce samples were processed with an immunomagnetic separation step before the qPCR assay. All retail lettuce samples were negative for E. coli O157:H7 when tested with the R.A.P.I.D. LT qPCR targeting a region of the O-antigen, and only two (0.28%) were positive for the eae gene when tested with LightCycler qPCR. On Petrifilm, coliform counts of most lettuce samples (96.4%) were between <101 and 103 CFU/g, and E. coli counts for nearly all lettuce samples (98.2%) were <101 CFU/g. No seasonal trend in coliform and E. coli counts was observed throughout the examination period nor was a difference in coliform counts observed between packaged and nonpackaged lettuce heads. These results contribute to the limited recorded data and understanding of microbial contamination of whole romaine lettuce heads purchased from retail locations, specifically revealing the absence of E. coli O157:H7 and low levels of contamination with coliforms and other E. coli strains.

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