Penetration of Escherichia coli O157:H7 into Lettuce Tissues as Affected by Inoculum Size and Temperature and the Effect of Chlorine Treatment on Cell Viability

2000 ◽  
Vol 63 (4) ◽  
pp. 434-440 ◽  
Author(s):  
KAZUE TAKEUCHI ◽  
JOSEPH F. FRANK
Keyword(s):  
Escherichia Coli ◽  
Cell Viability ◽  
Fluorescein Isothiocyanate ◽  
Inoculum Size ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Cut Edge ◽  
Log Reduction ◽  
Iceberg Lettuce ◽  
Effect Of Chlorine

Penetration of Escherichia coli O157:H7 into iceberg lettuce tissues and the effect of chlorine treatment on cell viability were evaluated. Attachment of different inoculum levels (109, 108, and 107 CFU/ml) was examined by determining the number of cells at the surface and the cut edge of lettuce leaves (2 by 2 cm). E. coli O157:H7 attached preferentially to cut edges at all inoculum levels, with greater attachment per cm2 of lettuce at higher inoculum levels. A longer attachment time allowed more cells to attach at both sites. Immunostaining with a fluorescein isothiocyanate–labeled antibody revealed that cells penetrated into lettuce leaves from cut edges. Cells showed greater penetration when lettuce was held at 4°C compared with 7, 25, or 37°C and were detected at an average of 73.5 ± 16.0 μm below the surfaces of cut tissues. Penetrating cells were mostly found at the junction of lettuce cells. The viability of attached cells after treatment with 200 mg/liter (200 ppm) of free chlorine for 5 min was examined by plating on tryptic soy agar and by a nalidixic acid elongation method. Although chlorine treatment caused significant reduction in attachment (0.7- and 1.0-log reduction at surfaces and cut edges, respectively), cells remained attached at high numbers (7.9 and 8.1 log CFU/cm2 at surfaces and cut edges, respectively). Elongated cells were observed in stomata and within the tissues of the lettuce, indicating they were protected from contact with chlorine.

Comparison of the Attachment of Escherichia coli O157:H7, Listeria monocytogenes, Salmonella Typhimurium, and Pseudomonas fluorescens to Lettuce Leaves

2000 ◽  
Vol 63 (10) ◽  
pp. 1433-1437 ◽  
Author(s):  
KAZUE TAKEUCHI ◽  
CLAUDIA M. MATUTE ◽  
ASHRAF N. HASSAN ◽  
JOSEPH F. FRANK
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Salmonella Typhimurium ◽  
Pseudomonas Fluorescens ◽  
Fluorescein Isothiocyanate ◽  
Plate Count ◽  
Confocal Scanning Laser Microscopy ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Cut Edge

Attachment of Escherichia coli O157:H7, Listeria monocytogenes, Salmonella Typhimurium, and Pseudomonas fluorescens on iceberg lettuce was evaluated by plate count and confocal scanning laser microscopy (CSLM). Attachment of each microorganism (∼108 CFU/ml) on the surface and the cut edge of lettuce leaves was determined. E. coli O157:H7 and L. monocytogenes attached preferentially to cut edges, while P. fluorescens attached preferentially to the intact surfaces. Differences in attachment at the two sites were greatest with L. monocytogenes. Salmonella Typhimurium attached equally to the two sites. At the surface, P. fluorescens attached in greatest number, followed by E. coli O157:H7, L. monocytogenes, and Salmonella Typhimurium. Attached microorganisms on lettuce were stained with fluorescein isothiocyanate and visualized by CSLM. Images at the surface and the cut edge of lettuce confirmed the plate count data. In addition, microcolony formation by P. fluorescens was observed on the lettuce surface. Some cells of each microorganism at the cut edge were located within the lettuce tissues, indicating that penetration occurred from the cut edge surface. The results of this study indicate that different species of microorganisms attach differently to lettuce structures, and CSLM can be successfully used to detect these differences.

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.

Penetration of Escherichia coli O157:H7 into Lettuce as Influenced by Modified Atmosphere and Temperature

2001 ◽  
Vol 64 (11) ◽  
pp. 1820-1823 ◽  
Author(s):  
KAZUE TAKEUCHI ◽  
ASHRAF N. HASSAN ◽  
JOSEPH F. FRANK
Keyword(s):  
Escherichia Coli ◽  
Oxygen Concentration ◽  
Respiration Rate ◽  
Atmospheric Oxygen ◽  
Fluorescein Isothiocyanate ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Iceberg Lettuce ◽  
Effects Of Temperature ◽  
Incubation Temperatures

The effects of temperature and atmospheric oxygen concentration on the respiration rate of iceberg lettuce and Escherichia coli O157:H7 cells attachment to and penetration into damaged lettuce tissues were evaluated. Respiration rate of lettuce decreased as the temperature was reduced from 37 to 10°C. Reducing the temperature further to 4°C did not affect the respiration rate of lettuce. Respiration rate was also reduced by lowering the atmospheric oxygen concentration. Lettuce was submerged in E. coli O157:H7 inoculum at 4, 10, 22, or 37°C under 21 or 2.7% oxygen. Attachment and penetration of E. coli O157:H7 were not related to the respiration rate. The greatest numbers of E. coli O157:H7 cells attached to damaged lettuce tissues at 22°C at both oxygen concentrations. More cells were attached under 21% oxygen than under 2.7% oxygen at each temperature, but this difference was small. Penetration of E. coli O157:H7 into lettuce tissue was determined by immunostaining with a fluorescein isothiocyanate-labeled antibody. Under 21% oxygen, E. coli O157:H7 cells showed greatest penetration when lettuce was held at 4°C, compared to 10, 22, or 37°C, and were detected at an average of 101 μm below the surfaces of cut tissues. However, under 2.7% oxygen, there were no differences in degree of penetration among four incubation temperatures. The degree of E. coli O157:H7 penetration into lettuce tissue at 4 or 22°C was greater under 21% oxygen than under 2.7% oxygen; however, no difference was observed at 37°C. Conditions that promote pathogen penetration into tissue could decrease the effectiveness of decontamination treatments.

Inactivation of Escherichia coli O157:H7 on Lettuce, Using Low-Energy X-Ray Irradiation

2010 ◽  
Vol 73 (3) ◽  
pp. 547-551 ◽  
Author(s):  
SANGHYUP JEONG ◽  
BRADLEY P. MARKS ◽  
ELLIOT T. RYSER ◽  
SCOTT R. MOOSEKIAN
Keyword(s):  
Escherichia Coli ◽  
Microbial Inactivation ◽  
Low Energy ◽  
Escherichia Coli O157 ◽  
X Ray ◽  
E Coli ◽  
Log Reduction ◽  
Leafy Greens ◽  
Iceberg Lettuce ◽  
Dose Levels

Low-energy X-ray irradiation was assessed as a means of eliminating Escherichia coli O157:H7 on lettuce. Round-cut iceberg lettuce samples (2.54-cm diameter) were dip or spot inoculated with a three-strain cocktail of E. coli O157:H7, stored for 24 h at 4°C, and then irradiated at four dose levels up to 0.25 kGy using a prototype low-energy (70 kV) X-ray irradiator. E. coli O157:H7 survivors were quantified by plating on sorbitol MacConkey agar containing cefixime and tellurite. Dip inoculation yielded a D10-value of 0.040 ± 0.001 kGy, which is 3.4 times lower than a previously reported value of 0.136 kGy using gamma radiation. The D10-value for E. coli O157:H7 on spot-inoculated samples was 0.078 ± 0.008 kGy, which is about twice that of dip-inoculated samples. When 10 stacked leaves were irradiated from both sides, a dose of 0.2 kGy was achieved at the center of the stack with a surface dose of 1 kGy, corresponding to a ~5-log reduction of E. coli O157:H7 at the center of the stack. Based on these findings, low-energy X-ray irradiation appears to be a promising microbial inactivation strategy for leafy greens and potentially for other types of fresh produce.

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.

Reduction of Escherichia coli O157:H7 Counts on Whole Fresh Apples by Treatment with Sanitizers

2000 ◽  
Vol 63 (6) ◽  
pp. 703-708 ◽  
Author(s):  
MARCY A. WISNIEWSKY ◽  
BONITA A. GLATZ ◽  
MARK L. GLEASON ◽  
CHERYLL A. REITMEIER
Keyword(s):  
Escherichia Coli ◽  
Chlorine Dioxide ◽  
Phosphate Buffer ◽  
Phosphate Buffer Solution ◽  
Escherichia Coli O157 ◽  
Peroxyacetic Acid ◽  
Water Control ◽  
Buffer Solution ◽  
E Coli ◽  
Log Reduction

The objectives of this study were to determine if washing of whole apples with solutions of three different sanitizers (peroxyacetic acid, chlorine dioxide, or a chlorine-phosphate buffer solution) could reduce a contaminating nonpathogenic Escherichia coli O157:H7 population by 5 logs and at what sanitizer concentration and wash time such a reduction could be achieved. Sanitizers were tested at 1, 2, 4, 8, and 16 times the manufacturer's recommended concentration at wash times of 5, 10, and 15 min. Whole, sound Braeburn apples were inoculated with approximately 1 × 108 or 7 × 106 CFU per apple, stored for 24 h, then washed with sterile water (control) or with sanitizers for the prescribed time. Recovered bacteria were enumerated on trypticase soy agar. Washing with water alone reduced the recoverable population by almost 2 logs from the starting population; this can be attributed to physical removal of organisms from the apple surface. No sanitizer, when used at the recommended concentration, reduced the recovered E. coli population by 5 logs under the test conditions. The most effective sanitizer, peroxyacetic acid, achieved a 5-log reduction when used at 2.1 to 14 times its recommended concentration, depending on the length of the wash time. The chlorine-phosphate buffer solution reduced the population by 5 logs when used at 3 to 15 times its recommended concentration, depending on wash time. At no concentration or wash time tested did chlorine dioxide achieve the 5-log reduction.

Inactivation of Escherichia coli O157:H7, Salmonella enterica Serotype Enteritidis, and Listeria monocytogenes on Lettuce by Hydrogen Peroxide and Lactic Acid and by Hydrogen Peroxide with Mild Heat

2002 ◽  
Vol 65 (8) ◽  
pp. 1215-1220 ◽  
Author(s):  
CHIA-MIN LIN ◽  
SARAH S. MOON ◽  
MICHAEL P. DOYLE ◽  
KAY H. McWATTERS
Keyword(s):  
Escherichia Coli ◽  
Hydrogen Peroxide ◽  
Lactic Acid ◽  
Listeria Monocytogenes ◽  
Salmonella Enterica ◽  
Salmonella Enteritidis ◽  
Sensory Characteristics ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Log Reduction

Iceberg lettuce is a major component in vegetable salad and has been associated with many outbreaks of foodborne illnesses. In this study, several combinations of lactic acid and hydrogen peroxide were tested to obtain effective antibacterial activity without adverse effects on sensory characteristics. A five-strain mixture of Escherichia coli O157:H7, Salmonella enterica serotype Enteritidis, and Listeria monocytogenes was inoculated separately onto fresh-cut lettuce leaves, which were later treated with 1.5% lactic acid plus 1.5% hydrogen peroxide (H2O2) at 40°C for 15 min, 1.5% lactic acid plus 2% H2O2 at 22°C for 5 min, and 2% H2O2 at 50°C for 60 or 90 s. Control lettuce leaves were treated with deionized water under the same conditions. A 4-log reduction was obtained for lettuce treated with the combinations of lactic acid and H2O2 for E. coli O157:H7 and Salmonella Enteritidis, and a 3-log reduction was obtained for L. monocytogenes. However, the sensory characteristics of lettuce were compromised by these treatments. The treatment of lettuce leaves with 2% H2O2 at 50°C was effective not only in reducing pathogenic bacteria but also in maintaining good sensory quality for up to 15 days. A ≤4-log reduction of E. coli O157:H7 and Salmonella Enteritidis was achieved with the 2% H2O2 treatment, whereas a 3-log reduction of L. monocytogenes was obtained. There was no significant difference (P > 0.05) between pathogen population reductions obtained with 2% H2O2 with 60- and 90-s exposure times. Hydrogen peroxide residue was undetectable (the minimum level of sensitivity was 2 ppm) on lettuce surfaces after the treated lettuce was rinsed with cold water and centrifuged with a salad spinner. Hence, the treatment of lettuce with 2% H2O2 at 50°C for 60 s is effective in initially reducing substantial populations of foodborne pathogens and maintaining high product quality.

Inactivation of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella in Cranberry, Lemon, and Lime Juice Concentrates

2003 ◽  
Vol 66 (9) ◽  
pp. 1637-1641 ◽  
Author(s):  
MARA C. L. NOGUEIRA ◽  
OMAR A. OYARZÁBAL ◽  
DAVID E. GOMBAS
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Pathogenic Bacteria ◽  
Antimicrobial Properties ◽  
Escherichia Coli O157 ◽  
Bacterial Inactivation ◽  
Lime Juice ◽  
E Coli ◽  
Log Reduction ◽  
C 32

The production of thermally concentrated fruit juices uses temperatures high enough to achieve at least a 5-log reduction of pathogenic bacteria that can occur in raw juice. However, the transportation and storage of concentrates at low temperatures prior to final packaging is a common practice in the juice industry and introduces a potential risk for postconcentration contamination with pathogenic bacteria. The present study was undertaken to evaluate the likelihood of Escherichia coli O157: H7, Listeria monocytogenes and Salmonella surviving in cranberry, lemon, and lime juice concentrates at or above temperatures commonly used for transportation or storage of these concentrates. This study demonstrates that cranberry, lemon, and lime juice concentrates possess intrinsic antimicrobial properties that will eliminate these bacterial pathogens in the event of postconcentration recontamination. Bacterial inactivation was demonstrated under all conditions; at least 5-log Salmonella inactivation was consistently demonstrated at −23°C (−10°F), at least 5-log E. coli O157:H7 inactivation was consistently demonstrated at −11°C (12°F), and at least 5-log L. monocytogenes inactivation was consistently demonstrated at 0°C (32°F).

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.

Inactivation of Pathogenic Bacteria by Cucumber Volatiles (E,Z)-2,6-Nonadienal and (E)-2-Nonenal†

2004 ◽  
Vol 67 (5) ◽  
pp. 1014-1016 ◽  
Author(s):  
M. J. CHO ◽  
R. W. BUESCHER ◽  
M. JOHNSON ◽  
M. JANES
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Salmonella Typhimurium ◽  
Bactericidal Activity ◽  
Pathogenic Bacteria ◽  
Brain Heart Infusion ◽  
Escherichia Coli O157 ◽  
Infusion Solution ◽  
E Coli ◽  
Log Reduction

The effects of (E,Z)-2,6-nonadienal (NDE) and (E)-2-nonenal (NE) on Bacillus cereus, Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella Typhimurium were investigated. A suspension of each organism of 6 to 9 log CFU/ml was incubated for 1 h at 37° C in brain heart infusion solution that contained 0 to 500 or 1,000 ppm of NDE or NE. Depending on concentration, exposure to either NDE or NE caused a reduction in CFU of each organism. Treatment with 250 and 500 ppm NDE completely eliminated viable B. cereus and Salmonella Typhimurium cells, respectively. L. monocytogenes was the most resistant to NDE, showing only about a 2-log reduction from exposure to 500 ppm for 1 h. Conversely, this concentration of NDE caused a 5.8-log reduction in E. coli O157:H7 cells. NE was also effective in inactivating organisms listed above. A higher concentration of NE, 1,000 ppm, was required to kill E. coli O157:H7, L. monocytogenes, or Salmonella Typhimurium compared with NDE. In conclusion, both NDE and NE demonstrated an apparent bactericidal activity against these pathogens.

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