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 ◽  
...  

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.

2017 ◽  
Vol 80 (5) ◽  
pp. 792-798 ◽  
Author(s):  
Jennifer A. Chase ◽  
Edward R. Atwill ◽  
Melissa L. Partyka ◽  
Ronald F. Bond ◽  
David Oryang

ABSTRACT A field trial was conducted in July 2011 to quantify the inactivation rate of Escherichia coli O157:H7 when mixed with fecal slurry and applied to romaine lettuce leaves. Lettuce was grown under commercial conditions in Salinas Valley, CA. One-half milliliter of rabbit fecal slurry, containing 6.3 × 107 CFU of E. coli O157:H7, was inoculated onto the upper (adaxial) surface of a lower leaf on 240 heads of lettuce within 30 min after a 2.5-h irrigation event. Forty-eight romaine lettuce heads were collected per event at 2.5 h (day 0.1), 19.75 h (day 0.8), 43.25 h (day 1.8), 67.25 h (day 2.8), and 91.75 h (day 3.8) postinoculation and were analyzed for the concentration of E. coli O157:H7 (Ct). E. coli O157:H7 was detected on 100% of collected heads in concentrations ranging from 340 to 3.40 × 1010 most probable number (MPN) per head. Enumeration data indicate substantial growth of E. coli O157:H7 postinoculation (2.5 h), leading to elevated concentrations, 1 to 3 log above the starting inoculum concentration (Co). By the end of the 92-h trial, we observed a net 0.8-log mean reduction of E. coli O157:H7 compared with Co; however, after accounting for the substantial bacterial growth, there was an overall 2.3-log reduction by the final sampling event (92 h). On the basis of two different regression models that used either the raw data for Ct or log-transformed values of Ct/Co during the period 2.5 to 91.75 h postinoculation, there was an estimated 76 to 80% reduction per day in bacterial counts; however, more accurate predictions of MPN per head of lettuce were generated by using non–log-transformed values of Ct. This study provides insight into the survival of E. coli O157:H7 transferred via splash from a contaminated fecal source onto produce during irrigation. Moreover, these findings can help generate inactivation times following a potential contamination incident.


2008 ◽  
Vol 71 (2) ◽  
pp. 252-256 ◽  
Author(s):  
JIN KYUNG KIM ◽  
MARK A. HARRISON

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.


2010 ◽  
Vol 73 (2) ◽  
pp. 212-220 ◽  
Author(s):  
ROWAIDA K. KHALIL ◽  
JOSEPH F. FRANK

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.


2009 ◽  
Vol 72 (7) ◽  
pp. 1553-1559 ◽  
Author(s):  
CHRISTOPHER G. THEOFEL ◽  
LINDA J. HARRIS

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.


2011 ◽  
Vol 74 (9) ◽  
pp. 1422-1433 ◽  
Author(s):  
CHARLES C. DODD ◽  
MICHAEL W. SANDERSON ◽  
MEGAN E. JACOB ◽  
DAVID G. RENTER

Field studies evaluating the effects of multiple concurrent preharvest interventions for Escherichia coli O157 are logistically and economically challenging; however, modeling techniques may provide useful information on these effects while also identifying crucial information gaps that can guide future research. We constructed a risk assessment model with data obtained from a systematic search of scientific literature. Parameter distributions were incorporated into a stochastic Monte Carlo modeling framework to examine the impacts of different combinations of preharvest and harvest interventions for E. coli O157 on the risk of beef carcass contamination. We estimated the risk of E. coli O157 carcass contamination conditional on preharvest fecal prevalence estimates, inclusion of feed additive(s) in the diet, vaccination for E. coli O157, transport and lairage effects, hide intervention(s), and carcass intervention(s). Prevalence parameters for E. coli O157 were assumed to encompass potential effects of concentration; therefore, concentration effects were not specifically evaluated in this study. Sensitivity analyses revealed that fecal prevalence, fecal-to-hide transfer, hide-to-carcass transfer, and carcass intervention efficacy significantly affected the risk of carcass contamination (correlation coefficients of 0.37, 0.56, 0.58, and −0.29, respectively). The results indicated that combinations of preharvest interventions may be particularly important for supplementing harvest interventions during periods of higher variability in fecal shedding prevalence (i.e., summer). Further assessments of the relationships among fecal prevalence and concentration, hide contamination, and subsequent carcass contamination are needed to further define risks and intervention impacts for E. coli O157 contamination of beef.


2012 ◽  
Vol 75 (3) ◽  
pp. 480-487 ◽  
Author(s):  
GREG BEZANSON ◽  
PASCAL DELAQUIS ◽  
SUSAN BACH ◽  
ROBIN McKELLAR ◽  
ED TOPP ◽  
...  

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.


2020 ◽  
Vol 83 (8) ◽  
pp. 1444-1462 ◽  
Author(s):  
GENEVIÈVE COULOMBE ◽  
ANGELA CATFORD ◽  
AMALIA MARTINEZ-PEREZ ◽  
ENRICO BUENAVENTURA

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


PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e6591 ◽  
Author(s):  
Jennifer A. Chase ◽  
Melissa L. Partyka ◽  
Ronald F. Bond ◽  
Edward R. Atwill

Field trials were conducted in July–August and October 2012 to quantify the inactivation rate of Escherichia coli O157:H7 when mixed with fecal slurry and applied to romaine lettuce leaves. Lettuce was grown under commercial conditions in Salinas Valley, California. One-half milliliter of rabbit, chicken, or pig fecal slurry, containing an average of 4.05 × 107 CFU E. coli O157:H7 (C0), was inoculated onto the upper (adaxial) surface of a lower leaf on 288 heads of lettuce per trial immediately following a 2.5 h irrigation event. To estimate the bacterial inactivation rate as a function of time, fecal matrix, irrigation and seasonal climate effects, sets of lettuce heads (n = 28) were sampled each day over 10 days and the concentration of E. coli O157:H7 (Ct) determined. E. coli O157:H7 was detected on 100% of heads during the 10-day duration, with concentrations ranging from ≤340 MPN/head (∼5-log reduction) to >3.45 × 1012 MPN/head (∼5-log growth). Relative to C0, on day 10 (Ct = 12) we observed an overall 2.6-log and 3.2-log mean reduction of E. coli O157:H7 in July and October, respectively. However, we observed relative maximum concentrations due to bacterial growth on day 6 (maximum Ct = 8) apparently stimulated by foliar irrigation on day 5. From this maximum there was a mean 5.3-log and 5.1-log reduction by day 10 (Ct = 12) for the July and October trials, respectively. This study provides insight into the inactivation and growth kinetics of E. coli O157:H7 on romaine lettuce leaves under natural field conditions. This study provides evidence that harvesting within 24 h post irrigation has the potential to increase the concentration of E. coli O157:H7 contamination, if present on heads of romaine lettuce; foliar irrigation can temporarily stimulate substantial regrowth of E. coli O157:H7.


2012 ◽  
Vol 75 (12) ◽  
pp. 2208-2212 ◽  
Author(s):  
PETER M. A. TOIVONEN ◽  
CHANGWEN LU ◽  
SUSAN BACH ◽  
PASCAL DELAQUIS

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.


2016 ◽  
Vol 80 (1) ◽  
pp. 35-43 ◽  
Author(s):  
SEA C. MIN ◽  
SI HYEON ROH ◽  
GLENN BOYD ◽  
JOSEPH E. SITES ◽  
JOSEPH UKNALIS ◽  
...  

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.


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