Influence of Curli Expression by Escherichia coli O157:H7 on the Cell's Overall Hydrophobicity, Charge, and Ability To Attach to Lettuce

2007 ◽  
Vol 70 (6) ◽  
pp. 1339-1345 ◽  
Author(s):  
RENEE R. BOYER ◽  
SUSAN S. SUMNER ◽  
ROBERT C. WILLIAMS ◽  
MERLE D. PIERSON ◽  
DAVID L. POPHAM ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Surface Charge ◽  
Extracellular Proteins ◽  
Escherichia Coli O157 ◽  
Fruit And Vegetable ◽  
E Coli ◽  
Iceberg Lettuce ◽  
Escherichia Coli Cells ◽  
Abiotic Surfaces ◽  
Plant Surfaces

Curli fibers are produced by some Escherichia coli cells in response to environmental stimuli. These extracellular proteins enhance the cell's ability to form biofilms on various abiotic surfaces. E. coli O157:H7 cells readily attach to a variety of fruit and vegetable surfaces. It is not known whether the expression of curli influences the cell's ability to attach to produce surfaces. In this study, the effect of curli expression on the cell's overall hydrophobicity, charge, and ability to attach to cut and whole iceberg lettuce surfaces was examined. All strains, regardless of curli expression, attached preferentially to the cut edges of lettuce (P < 0.05). The curli-producing cells of E. coli O157:H7 strain E0018 attached in significantly greater numbers to both cut and whole lettuce pieces than did the non–curli-producing E0018 cells (P < 0.05); however, no significant attachment differences were observed between the curli-producing and non–curli-producing cells of E. coli O157:H7 strains 43894 and 43895. All curli-producing E. coli O157:H7 strains were significantly more hydrophobic (P < 0.01); however, no association between the cells' hydrophobic characteristics and lettuce attachment was observed. Overall surface charge of the cells did not differ among strains or curli phenotypes. Results indicate that overall hydrophobicity and cell charge in E. coli O157:H7 strains do not influence attachment to iceberg lettuce surfaces. The presence of curli may not have any influence on attachment of E. coli O157:H7 cells to produce items. Additional factors may influence the attachment of E. coli O157:H7 to plant surfaces and should be further examined.

Tracking an Escherichia coli O157:H7–Contaminated Batch of Leafy Greens through a Pilot-Scale Fresh-Cut Processing Line

2014 ◽  
Vol 77 (9) ◽  
pp. 1487-1494 ◽  
Author(s):  
ANNEMARIE L. BUCHHOLZ ◽  
GORDON R. DAVIDSON ◽  
BRADLEY P. MARKS ◽  
EWEN C. D. TODD ◽  
ELLIOT T. RYSER
Keyword(s):  
Escherichia Coli ◽  
Membrane Filtration ◽  
Fluorescent Protein ◽  
Pilot Scale ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Leafy Greens ◽  
Iceberg Lettuce ◽  
Processing Line ◽  
Fresh Cut

Cross-contamination of fresh-cut leafy greens with residual Escherichia coli O157:H7–contaminated product during commercial processing was likely a contributing factor in several recent multistate outbreaks. Consequently, radicchio was used as a visual marker to track the spread of the contaminated product to iceberg lettuce in a pilot-scale processing line that included a commercial shredder, step conveyor, flume tank, shaker table, and centrifugal dryer. Uninoculated iceberg lettuce (45 kg) was processed, followed by 9.1 kg of radicchio (dip inoculated to contain a four-strain, green fluorescent protein–labeled nontoxigenic E. coli O157:H7 cocktail at 106 CFU/g) and 907 kg (2,000 lb) of uninoculated iceberg lettuce. After collecting the lettuce and radicchio in about 40 bags (~22.7 kg per bag) along with water and equipment surface samples, all visible shreds of radicchio were retrieved from the bags of shredded product, the equipment, and the floor. E. coli O157:H7 populations were quantified in the lettuce, water, and equipment samples by direct plating with or without prior membrane filtration on Trypticase soy agar containing 0.6% yeast extract and 100 ppm of ampicillin. Based on triplicate experiments, the weight of radicchio in the shredded lettuce averaged 614.9 g (93.6%), 6.9 g (1.3%), 5.0 g (0.8%), and 2.8 g (0.5%) for bags 1 to 10, 11 to 20, 21 to 30, and 31 to 40, respectively, with mean E. coli O157:H7 populations of 1.7, 1.2, 1.1, and 1.1 log CFU/g in radicchio-free lettuce. After processing, more radicchio remained on the conveyor (9.8 g; P < 0.05), compared with the shredder (8.3 g), flume tank (3.5 g), and shaker table (0.1 g), with similar E. coli O157:H7 populations (P > 0.05) recovered from all equipment surfaces after processing. These findings clearly demonstrate both the potential for the continuous spread of contaminated lettuce to multiple batches of product during processing and the need for improved equipment designs that minimize the buildup of residual product during processing.

Is There a Relation between the Microscopic Leaf Morphology and the Association of Salmonella and Escherichia coli O157:H7 with Iceberg Lettuce Leaves?

2016 ◽  
Vol 79 (10) ◽  
pp. 1784-1788 ◽  
Author(s):  
INGE VAN der LINDEN ◽  
MARKUS ERIKSSON ◽  
MIEKE UYTTENDAELE ◽  
FRANK DEVLIEGHERE
Keyword(s):  
Escherichia Coli ◽  
Stomatal Density ◽  
Fresh Produce ◽  
Morphological Structure ◽  
Bacterial Attachment ◽  
Escherichia Coli O157 ◽  
Stages Of Development ◽  
E Coli ◽  
Iceberg Lettuce ◽  
Morphological Differences

ABSTRACT To prevent contamination of fresh produce with enteric pathogens, more insight into mechanisms that may influence the association of these pathogens with fresh produce is needed. In this study, Escherichia coli O157:H7 and Salmonella were chosen as model pathogens, and fresh cut iceberg lettuce was chosen as a model fresh produce type. The morphological structure of iceberg lettuce leaves (stomatal density and length of cell margins per leaf area) was quantified by means of leaf peels and light microscopy of leaves at different stages of development (outer, middle, and inner leaves of the crop) on both leaf sides (abaxial and adxial) and in three leaf regions (top, center, and bottom). The morphology of the top region of the leaves was distinctly different from that of the center and base, with a significantly higher stomatal density (up to five times more stomata), different cell shape, and longer cell margins (two to three times longer). Morphological differences between the same regions of the leaves at different stages of development were smaller or nonsignificant. An attachment assay with two attenuated E. coli O157:H7 strains (84-24h11-GFP and BRMSID 188 GFP) and two Salmonella strains (serovars Thompson and Typhimurium) was performed on different regions of the middle leaves. Our results confirmed earlier reports that these pathogens have a higher affinity for the base of the lettuce leaf than the top. Differences of up to 2.12 log CFU/g were seen (E. coli O157:H7 86-24h11-GFP). Intermediate attachment occurred in the central region. The higher incidence of preferential bacterial attachment sites such as stomata and cell margins or grooves could not explain the differences observed in the association of the tested pathogens with different regions of iceberg lettuce leaves.

Efficacy of Commercial Produce Sanitizers against Nontoxigenic Escherichia coli O157:H7 during Processing of Iceberg Lettuce in a Pilot-Scale Leafy Green Processing Line

2013 ◽  
Vol 76 (11) ◽  
pp. 1838-1845 ◽  
Author(s):  
GORDON R. DAVIDSON ◽  
ANNEMARIE L. BUCHHOLZ ◽  
ELLIOT T. RYSER
Keyword(s):  
Escherichia Coli ◽  
Fluorescent Protein ◽  
Pilot Scale ◽  
Wash Water ◽  
Escherichia Coli O157 ◽  
Cross Contamination ◽  
E Coli ◽  
Iceberg Lettuce ◽  
Processing Line ◽  
Green Fluorescent

Chemical sanitizers are routinely used during commercial flume washing of fresh-cut leafy greens to minimize cross-contamination from the water. This study assessed the efficacy of five commercial sanitizer treatments against Escherichia coli O157:H7 on iceberg lettuce, in wash water, and on equipment during simulated commercial production in a pilot-scale processing line. Iceberg lettuce (5.4 kg) was inoculated to contain 106 CFU/g of a four-strain cocktail of nontoxigenic, green fluorescent protein–labeled, ampicillin-resistant E. coli O157:H7 and processed after 1 h of draining at ~22°C. Lettuce was shredded using a commercial slicer, step-conveyed to a flume tank, washed for 90 s using six different treatments (water alone, 50 ppm of peroxyacetic acid, 50 ppm of mixed peracid, or 50 ppm of available chlorine either alone or acidified to pH 6.5 with citric acid [CA] or T-128), and then dried using a shaker table and centrifugal dryer. Various product (25-g) and water (50-ml) samples collected during processing along with equipment surface samples (100 cm2) from the flume tank, shaker table, and centrifugal dryer were homogenized in neutralizing buffer and plated on tryptic soy agar. During and after iceberg lettuce processing, none of the sanitizers were significantly more effective (P ≤ 0.05) than water alone at reducing E. coli O157:H7 populations on lettuce, with reductions ranging from 0.75 to 1.4 log CFU/g. Regardless of the sanitizer treatment used, the centrifugal dryer surfaces yielded E. coli O157:H7 populations of 3.49 to 4.98 log CFU/100 cm2. Chlorine, chlorine plus CA, and chlorine plus T-128 were generally more effective (P ≤ 0.05) than the other treatments, with reductions of 3.79, 5.47, and 5.37 log CFU/ml after 90 s of processing, respectively. This indicates that chlorine-based sanitizers will likely prevent wash water containing low organic loads from becoming a vehicle for cross-contamination.

Fate of Shiga Toxin–Producing O157:H7 and Non-O157:H7 Escherichia coli Cells within Blade-Tenderized Beef Steaks after Cooking on a Commercial Open-Flame Gas Grill†

2012 ◽  
Vol 75 (1) ◽  
pp. 62-70 ◽  
Author(s):  
JOHN B. LUCHANSKY ◽  
ANNA C. S. PORTO-FETT ◽  
BRADLEY A. SHOYER ◽  
JEFFREY E. CALL ◽  
WAYNE SCHLOSSER ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Phase I ◽  
Phase Ii ◽  
Shiga Toxin ◽  
Escherichia Coli O157 ◽  
Open Flame ◽  
E Coli ◽  
Escherichia Coli Cells ◽  
Beef Steaks ◽  
Uneven Heating

We compared the fate of cells of both Shiga toxin–producing Escherichia coli O157:H7 (ECOH) and Shiga toxin–producing non-O157:H7 E. coli (STEC) in blade-tenderized steaks after tenderization and cooking on a gas grill. In phase I, beef subprimal cuts were inoculated on the lean side with about 5.5 log CFU/g of a five-strain mixture of ECOH or STEC and then passed once through a mechanical blade tenderizer with the lean side facing up. In each of two trials, 10 core samples were removed from each of two tenderized subprimals and cut into six consecutive segments starting from the inoculated side. Ten total cores also were obtained from two nontenderized (control) subprimals, but only segment 1 (the topmost segment) was sampled. The levels of ECOH and STEC recovered from segment 1 were about 6.0 and 5.3 log CFU/g, respectively, for the control subprimals and about 5.7 and 5.0 log CFU/g, respectively, for the tenderized subprimals. However, both ECOH and STEC behaved similarly in terms of translocation, and cells of both pathogen cocktails were recovered from all six segments of the cores obtained from tenderized subprimals, albeit at lower levels in segments 2 to 6 than those found in segment 1. In phase II, steaks (2.54 and 3.81 cm thick) cut from tenderized subprimals were subsequently cooked (three steaks per treatment) on a commercial open-flame gas grill to internal temperatures of 48.9, 54.4, 60.0, 65.6, and 71.1°C. Regardless of temperature or thickness, we observed 2.0- to 4.1-log and 1.5- to 4.5-log reductions in ECOH and STEC levels, respectively. Both ECOH and STEC behaved similarly in response to heat, in that cooking eliminated significant numbers of both pathogen types; however, some survivors were recovered due, presumably, to uneven heating of the blade-tenderized steaks.

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

Reduction of Escherichia coli O157:H7 on Produce by Use of Electrolyzed Water under Simulated Food Service Operation Conditions

2009 ◽  
Vol 72 (9) ◽  
pp. 1854-1861 ◽  
Author(s):  
PHILIPUS PANGLOLI ◽  
YEN-CON HUNG ◽  
LARRY R. BEUCHAT ◽  
C. HAROLD KING ◽  
ZHI-HUI ZHAO
Keyword(s):  
Escherichia Coli ◽  
Foodborne Pathogens ◽  
Fruits And Vegetables ◽  
Tap Water ◽  
Food Service ◽  
Escherichia Coli O157 ◽  
Electrolyzed Water ◽  
E Coli ◽  
Operation Conditions ◽  
Iceberg Lettuce

Treatment of fresh fruits and vegetables with electrolyzed water (EW) has been shown to kill or reduce foodborne pathogens. We evaluated the efficacy of EW in killing Escherichia coli O157:H7 on iceberg lettuce, cabbage, lemons, and tomatoes by using washing and/or chilling treatments simulating those followed in some food service kitchens. Greatest reduction levels on lettuce were achieved by sequentially washing with 14-A (amperage) acidic EW (AcEW) for 15 or 30 s followed by chilling in 16-A AcEW for 15 min. This procedure reduced the pathogen by 2.8 and 3.0 log CFU per leaf, respectively, whereas washing and chilling with tap water reduced the pathogen by 1.9 and 2.4 log CFU per leaf. Washing cabbage leaves for 15 or 30 s with tap water or 14-A AcEW reduced the pathogen by 2.0 and 3.0 log CFU per leaf and 2.5 to 3.0 log CFU per leaf, respectively. The pathogen was reduced by 4.7 log CFU per lemon by washing with 14-A AcEW and 4.1 and 4.5 log CFU per lemon by washing with tap water for 15 or 30 s. A reduction of 5.3 log CFU per lemon was achieved by washing with 14-A alkaline EW for 15 s prior to washing with 14-A AcEW for 15 s. Washing tomatoes with tap water or 14-A AcEW for 15 s reduced the pathogen by 6.4 and 7.9 log CFU per tomato, respectively. Application of AcEW using procedures mimicking food service operations should help minimize cross-contamination and reduce the risk of E. coli O157:H7 being present on produce at the time of consumption.

Bactericidal Activity of Isothiocyanate against Pathogens on Fresh Produce†

2000 ◽  
Vol 63 (1) ◽  
pp. 25-30 ◽  
Author(s):  
CHIA-MIN LIN ◽  
JEONGMOK KIM ◽  
WEN-XIAN DU ◽  
CHENG-I WEI
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Bactericidal Activity ◽  
Bactericidal Effect ◽  
Escherichia Coli O157 ◽  
Methyl Isothiocyanate ◽  
E Coli ◽  
Log Reduction ◽  
Iceberg Lettuce ◽  
Resistant Strains

The bactericidal activity of allyl and methyl isothiocyanate (AITC and MITC) was tested with a rifampicin-resistant strain of Salmonella Montevideo and streptomycin-resistant strains of Escherichia coli O157:H7 and Listeria monocytogenes Scott A. Iceberg lettuce inoculated with high (107 to 108 CFU/g) and low (103 to 104 CFU/g) concentrations of bacterial pathogens was treated with AITC and MITC in sealed containers at 4°C for 4 days. AITC showed stronger bactericidal activity than MITC against E. coli O157:H7 and Salmonella Montevideo, whereas MITC showed stronger activity against L. monocytogenes than E. coli O157:H7 and Salmonella Montevideo. Up to 8-log reduction occurred with E. coli O157:H7 and Salmonella Montevideo on lettuce following treatment with vapor generated from 400 μl of AITC for 2 and 4 days, respectively. AITC was used to treat tomatoes inoculated with Salmonella Montevideo on stem scars and skin and apples inoculated with E. coli O157:H7 on stem scars. The bactericidal effect of AITC varied with bacteria species and exposure time. Salmonella Montevideo inoculated on tomato skin was more sensitive to AITC than that on stem scars. Treatment with vapor generated from 500 μl of AITC caused an 8-log reduction in bacteria on tomato skin but only a 5-log reduction on tomato stem scars. The bactericidal activity of AITC was weaker for E. coli O157:H7 on apple stem scars; only a 3-log reduction in bacteria occurred when 600 μl of AITC was used.

Survival and Recovery of Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes on Lettuce and Parsley as Affected by Method of Inoculation, Time between Inoculation and Analysis, and Treatment with Chlorinated Water

2004 ◽  
Vol 67 (6) ◽  
pp. 1092-1103 ◽  
Author(s):  
MEGAN M. LANG ◽  
LINDA J. HARRIS ◽  
LARRY R. BEUCHAT
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Foodborne Pathogens ◽  
Escherichia Coli O157 ◽  
Water Control ◽  
Drying Time ◽  
E Coli ◽  
Iceberg Lettuce ◽  
Spray Inoculation ◽  
Number Of Cells

The effects of method for applying inoculum and of drying time after inoculation on survival and recovery of foodborne pathogens on iceberg lettuce and parsley were studied. Five-strain mixtures of Escherichia coli O157:H7, Salmonella, or Listeria monocytogenes were applied to lettuce and parsley by dip, spot, or spray inoculation methods. Inocula were dried for 2 h at 22°C or for 2 h at 22°C and then 22 h at 4°C before being treated with water (control) or chlorine (200 μg/ml). Significantly higher populations (CFU per lettuce or parsley sample) of E. coli O157:H7 and Salmonella (α = 0.05) were recovered from dip-inoculated produce than from spot- or spray-inoculated produce. This difference was attributed to larger numbers of cells adhering to lettuce and parsley subjected to dip inoculation. Populations of E. coli O157:H7 and Salmonella recovered from lettuce inoculated by spot and spray methods were not significantly different, but populations recovered from spot-inoculated parsley were significantly higher than those recovered from spray-inoculated parsley, even though the number of cells applied was the same. Significantly different numbers of L. monocytogenes were recovered from inoculated lettuce (dip > spray > spot); populations recovered from dip-inoculated parsley were significantly higher than those recovered from spot- or spray-inoculated parsley, which were not significantly different from each other. Populations of pathogens recovered from lettuce and parsley after drying inoculum for 2 h at 22°C were significantly higher than or equal to populations recovered after drying for 2 h at 22°C and then for 22 h at 4°C. Significant differences (water > chlorine) were observed in populations of all pathogens recovered from treated lettuce and parsley, regardless of inoculation method and drying time. It is recommended that spot inoculation with a drying time of 2 h at 22°C followed by 22 h at 4°C be used to determine the efficacy of chlorine and other sanitizers in killing foodborne pathogens on lettuce and parsley.

Survival and Growth of Escherichia coli O157:H7 Inoculated onto Cut Lettuce Before or After Heating in Chlorinated Water, Followed by Storage at 5 or 15°C

2001 ◽  
Vol 64 (3) ◽  
pp. 305-309 ◽  
Author(s):  
YUE LI ◽  
ROBERT E. BRACKETT ◽  
JINRU CHEN ◽  
LARRY R. BEUCHAT
Keyword(s):  
Escherichia Coli ◽  
Escherichia Coli O157 ◽  
Marked Influence ◽  
E Coli ◽  
Iceberg Lettuce ◽  
Survival And Growth ◽  
Fresh Cut ◽  
Subsequent Storage ◽  
Efficacy Of Treatment ◽  
Treatment Water

This study determined the effects of mild heat and chlorine treatment followed by storage for up to 18 days at 5°C or 7 days at 15°C on the survival and growth of Escherichia coli O157:H7 inoculated onto fresh-cut iceberg lettuce. The efficacy of treatment with 20 ppm chlorine in killing the pathogen on lettuce at 50°C was determined. Treatment of lettuce with 20 ppm chlorine at either 20 or 50°C did not result in significantly greater reductions in populations of E. coli O157:H7 compared to respective treatments in water without chlorine. The pathogen steadily decreased in viability on treated lettuce throughout subsequent storage at 5°C for 18 days. The population increased by 2.3 to 3.2 log10 CFU/g within 2 days, then continued to increase at a slower rate through 7 days of storage at 15°C. At 4 and 7 days, significantly (α = 0.05) higher populations were reached on lettuce that had been treated at 50°C, compared to respective samples that had been treated at 20°C, regardless of the presence of 20 ppm chlorine in the treatment water. Treatment of lettuce with 20 ppm chlorine at 50 or 20°C before or after inoculation with E. coli O157:H7 did not have a marked influence on behavior of the pathogen during subsequent storage at 5 or 15°C.

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