Fate of Escherichia coli O157:H7 in Manure Compost–Amended Soil and on Carrots and Onions Grown in an Environmentally Controlled Growth Chamber

2004 ◽  
Vol 67 (3) ◽  
pp. 574-578 ◽  
Author(s):  
MAHBUB ISLAM ◽  
JENNIE MORGAN ◽  
MICHAEL P. DOYLE ◽  
XIUPING JIANG
Keyword(s):  
Escherichia Coli ◽  
Fluorescent Protein ◽  
Sandy Loam ◽  
Growth Cycle ◽  
Growth Chamber ◽  
Controlled Growth ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Manure Compost ◽  
Amended Soil

Studies were done to determine the fate of Escherichia coli O157:H7 in manure compost–amended soil and on carrots and green onions grown in an environmentally controlled growth chamber. Commercial dairy cattle manure compost was inoculated with a five-strain mixture of green fluorescent protein–labeled E. coli O157:H7 at 107 CFU g−1 and mixed with unsterilized Tifton sandy loam soil at a ratio of 1:5. Baby carrot or green onion seedlings were planted into the manure compost–amended soil in pots, and soil samples surrounding the plant, edible carrot roots and onion bulb samples, and soil immediately beneath the roots were assayed for E. coli O157:H7 in triplicate at weekly intervals for the first 4 weeks, and every 2 weeks for the remainder of the plant growth cycle (up to 3 months). E. coli O157:H7 cell numbers decreased within 64 days by 3 log CFU/g in soil and soil beneath the roots of green onions and by more than 2 log CFU/g on onions. E. coli O157:H7 survived better during the production of carrots, with a 2.3-log CFU/g reduction in soil and a 1.7-log CFU/g reduction on carrots within 84 days. These results indicate that the type of plant grown is an important factor influencing the survival of E. coli O157:H7 both on the vegetable and in the soil in which the vegetable is grown.

Thermal Inactivation of Escherichia coli O157:H7 in Cow Manure Compost

2003 ◽  
Vol 66 (10) ◽  
pp. 1771-1777 ◽  
Author(s):  
XIUPING JIANG ◽  
JENNIE MORGAN ◽  
MICHAEL P. DOYLE
Keyword(s):  
Escherichia Coli ◽  
Green Fluorescent Protein ◽  
Thermal Inactivation ◽  
Fluorescent Protein ◽  
Escherichia Coli O157 ◽  
Cow Manure ◽  
E Coli ◽  
Manure Compost ◽  
D Values ◽  
Green Fluorescent

Rates of inactivation of a five-strain mixture of green fluorescent protein–labeled Escherichia coli O157:H7 in autoclaved and unautoclaved commercial cow manure compost with a moisture content of ca. 38% were determined at temperatures of 50, 55, 60, 65, and 70°C. Trypticase soy agar with ampicillin was determined to be the best medium for the enumeration of heat-injured and uninjured cells of green fluorescent protein–labeled E. coli O157:H7. The results obtained in this study revealed that in autoclaved compost, E. coli O157:H7 reductions of ca. 4 log CFU/g occurred within 8 h, 3 h, 15 min, 2 min, and <1 min at 50, 55, 60, 65, and 70°C, respectively. At 65 and 70°C, considerably less time was required to kill the pathogen in unautoclaved compost than in autoclaved compost. Decimal reduction times (D-values) for autoclaved compost at 50, 55, 60, 65, and 70°C were 137, 50.3, 4.1, 1.8, and 0.93 min, respectively, and D-values for unautoclaved compost at 50, 55, and 60°C were 135, 35.4, and 3.9 min, respectively. Considerable tailing was observed for inactivation curves, especially at 60, 65, and 70°C. These results are useful for identifying composting conditions that will reduce the risk of the transmission of E. coli O157:H7 to foods produced in the presence of animal fecal waste.

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.

A Novel Approach To Investigate the Uptake and Internalization of Escherichia coli O157:H7 in Spinach Cultivated in Soil and Hydroponic Medium†

2009 ◽  
Vol 72 (7) ◽  
pp. 1513-1520 ◽  
Author(s):  
MANAN SHARMA ◽  
DAVID T. INGRAM ◽  
JITENDRA R. PATEL ◽  
PATRICIA D. MILLNER ◽  
XIAOLIN WANG ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Plasmid Vector ◽  
Green Fluorescent Protein Gene ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Novel Approach ◽  
Efficient Uptake ◽  
Green Fluorescent

Internalization of Escherichia coli O157:H7 into spinach plants through root uptake is a potential route of contamination. ATn7-based plasmid vector was used to insert a green fluorescent protein gene into the attTn7 site in the E. coli chromosome. Three green fluorescent protein–labeled E. coli inocula were used: produce outbreak O157:H7 strains RM4407 and RM5279 (inoculum 1), ground beef outbreak O157:H7 strain 86-24h11 (inoculum 2), and commensal strain HS (inoculum 3). These strains were cultivated in fecal slurries and applied at ca. 103 or 107 CFU/g to pasteurized soils in which baby spinach seedlings were planted. No E. coli was recovered by spiral plating from surface-sanitized internal tissues of spinach plants on days 0, 7, 14, 21, and 28. Inoculum 1 survived at significantly higher populations (P < 0.05) in the soil than did inoculum 3 after 14, 21, and 28 days, indicating that produce outbreak strains of E. coli O157:H7 may be less physiologically stressed in soils than are nonpathogenic E. coli isolates. Inoculum 2 applied at ca. 107 CFU/ml to hydroponic medium was consistently recovered by spiral plating from the shoot tissues of spinach plants after 14 days (3.73 log CFU per shoot) and 21 days (4.35 log CFU per shoot). Fluorescent E. coli cells were microscopically observed in root tissues in 23 (21%) of 108 spinach plants grown in inoculated soils. No internalized E. coli was microscopically observed in shoot tissue of plants grown in inoculated soil. These studies do not provide evidence for efficient uptake of E. coli O157:H7 from soil to internal plant tissue.

Effect of Route of Introduction and Host Cultivar on the Colonization, Internalization, and Movement of the Human Pathogen Escherichia coli O157:H7 in Spinach

2009 ◽  
Vol 72 (7) ◽  
pp. 1521-1530 ◽  
Author(s):  
R. MITRA ◽  
E. CUESTA-ALONSO ◽  
A. WAYADANDE ◽  
J. TALLEY ◽  
S. GILLILAND ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Fluorescent Protein ◽  
Sprinkler Irrigation ◽  
Conclusive Evidence ◽  
Escherichia Coli O157 ◽  
Human Pathogens ◽  
Soil Infiltration ◽  
Fresh Tissue ◽  
E Coli ◽  
Over Time

Human pathogens can contaminate leafy produce in the field by various routes. We hypothesized that interactions between Escherichia coli O157:H7 and spinach are influenced by the route of introduction and the leaf microenvironment. E. coli O157:H7 labeled with green fluorescent protein was dropped onto spinach leaf surfaces, simulating bacteria-laden raindrops or sprinkler irrigation, and survived on the phylloplane for at least 14 days, with increasing titers and areas of colonization over time. The same strains placed into the rhizosphere by soil infiltration remained detectable on very few plants and in low numbers (102 to 106 CFU/g fresh tissue) that decreased over time. Stem puncture inoculations, simulating natural wounding, rarely resulted in colonization or multiplication. Bacteria forced into the leaf interior survived for at least 14 days in intercellular spaces but did not translocate or multiply. Three spinach cultivars with different leaf surface morphologies were compared for colonization by E. coli O157:H7 introduced by leaf drop or soil drench. After 2 weeks, cv. Bordeaux hosted very few bacteria. More bacteria were seen on cv. Space and were dispersed over an area of up to 0.3 mm2. The highest bacterial numbers were observed on cv. Tyee but were dispersed only up to 0.15 mm2, suggesting that cv. Tyee may provide protected niches or more nutrients or may promote stronger bacterial adherence. These findings suggest that the spinach phylloplane is a supportive niche for E. coli O157:H7, but no conclusive evidence was found for natural entry into the plant interior. The results are relevant for interventions aimed at minimizing produce contamination by human pathogens.

Persistence of Enterohemorrhagic Escherichia coli O157:H7 in Soil and on Leaf Lettuce and Parsley Grown in Fields Treated with Contaminated Manure Composts or Irrigation Water

2004 ◽  
Vol 67 (7) ◽  
pp. 1365-1370 ◽  
Author(s):  
MAHBUB ISLAM ◽  
MICHAEL P. DOYLE ◽  
SHARAD C. PHATAK ◽  
PATRICIA MILLNER ◽  
XIUPING JIANG
Keyword(s):  
Escherichia Coli ◽  
Irrigation Water ◽  
Poultry Manure ◽  
Dairy Manure ◽  
Enterohemorrhagic Escherichia Coli ◽  
Avirulent Strain ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Manure Compost ◽  
Contaminated Irrigation Water

Outbreaks of enterohemorrhagic Escherichia coli O157:H7 infections associated with lettuce and other leaf crops have occurred with increasing frequency in recent years. Contaminated manure and polluted irrigation water are probable vehicles for the pathogen in many outbreaks. In this study, the occurrence and persistence of E. coli O157:H7 in soil fertilized with contaminated poultry or bovine manure composts or treated with contaminated irrigation water and on lettuce and parsley grown on these soils under natural environmental conditions was determined. Twenty-five plots, each 1.8 by 4.6 m, were used for each crop, with five treatments (one without compost, three with each of the three composts, and one without compost but treated with contaminated water) and five replication plots for each treatment. Three different types of compost, PM-5 (poultry manure compost), 338 (dairy manure compost), and NVIRO-4 (alkaline-stabilized dairy manure compost), and irrigation water were inoculated with an avirulent strain of E. coli O157:H7. Pathogen concentrations were 107 CFU/g of compost and 105 CFU/ml of water. Contaminated compost was applied to soil in the field as a strip at 4.5 metric tons per hectare on the day before lettuce and parsley seedlings were transplanted in late October 2002. Contaminated irrigation water was applied only once on the plants as a treatment in five plots for each crop at the rate of 2 liters per plot 3 weeks after the seedlings were transplanted. E. coli O157:H7 persisted for 154 to 217 days in soils amended with contaminated composts and was detected on lettuce and parsley for up to 77 and 177 days, respectively, after seedlings were planted. Very little difference was observed in E. coli O157:H7 persistence based on compost type alone. E. coli O157:H7 persisted longer (by >60 days) in soil covered with parsley plants than in soil from lettuce plots, which were bare after lettuce was harvested. In all cases, E. coli O157:H7 in soil, regardless of source or crop type, persisted for >5 months after application of contaminated compost or irrigation water.

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.

scholarly journals Fate of Escherichia coli O157:H7 in Manure-Amended Soil

2002 ◽  
Vol 68 (5) ◽  
pp. 2605-2609 ◽  
Author(s):  
Xiuping Jiang ◽  
Jennie Morgan ◽  
Michael P. Doyle
Keyword(s):  
Escherichia Coli ◽  
Soil Temperature ◽  
Soil Microorganisms ◽  
Soil Samples ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Contributory Factors ◽  
Pathogen Populations ◽  
Antagonistic Interactions ◽  
Amended Soil

ABSTRACT Escherichia coli O157:H7 cells survived for up to 77, >226, and 231 days in manure-amended autoclaved soil held at 5, 15, and 21°C, respectively. Pathogen populations declined more rapidly in manure-amended unautoclaved soil under the same conditions, likely due to antagonistic interactions with indigenous soil microorganisms. E. coli O157:H7 cells were inactivated more rapidly in both autoclaved and unautoclaved soils amended with manure at a ratio of 1 part manure to 10 parts soil at 15 and 21°C than in soil samples containing dilute amounts of manure. The manure-to-soil ratio, soil temperature, and indigenous microorganisms of the soil appear to be contributory factors to the pathogen's survival in manure-amended soil.

scholarly journals Survival study of escherichia coli o157:h7 in aquatic system of Bangladesh

1970 ◽  
Vol 19 (2) ◽  
pp. 195-201
Author(s):  
Iqbal Kabir Jahid ◽  
Taslima Azad ◽  
Mohammed Ziaur Rahman ◽  
Anowara Begum ◽  
Sirajul Islam Khan ◽  
...  
Keyword(s):  
Waste Water ◽  
Escherichia Coli ◽  
Fluorescent Protein ◽  
Uv Light ◽  
Tap Water ◽  
Aquatic System ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Survival Study ◽  
Survival Pattern

The survival pattern of Escherichia coli O157:H7 was observed in laboratory microcosm with different sources of surface and drinking water using the green fluorescent protein (GFP) as a genetic marker. The water quality was monitored on the basis of bacteriological and physico?chemical parameters. The untreated and filtered water were inoculated with the genetically transformed E. coli O157:H7. The survival pattern was determined by drop plate method observing the green fluorescence of the E. coli O157:H7 colonies under UV light. The survival of E. coli O157:H7 decreased in most of the untreated saline and waste water and higher survival was observed in pond and tap water. The E. coli O157:H7 survived more than 23 days in tap and pond water and less than 20 days in sea, estuarine and waste water. The fluorescent microscopic findings revealed the VBNC state of E. coli O157:H7. The study conclusively proved that the better survival of E. coli O157: H7 depends on the quality of water. Key words: E. coli O157:H7; Aquatic system; Survival study DOI: http://dx.doi.org/10.3329/dujbs.v19i2.8964 DUJBS 2010; 19(2): 195-201

Factors Impacting the Regrowth of Escherichia coli O157:H7 in Dairy Manure Compost

2009 ◽  
Vol 72 (7) ◽  
pp. 1576-1584 ◽  
Author(s):  
JINKYUNG KIM ◽  
FENG LUO ◽  
XIUPING JIANG
Keyword(s):  
Escherichia Coli ◽  
Moisture Content ◽  
Room Temperature ◽  
Dairy Manure ◽  
Population Increase ◽  
Escherichia Coli O157 ◽  
Inoculum Level ◽  
Factors Affecting ◽  
E Coli ◽  
Manure Compost

The environmental variables affecting Escherichia coli O157:H7 regrowth in dairy manure compost were investigated. Factors evaluated were moisture content, strain variation, growth medium of inoculum, level of background microflora and inoculum, different days of composting, and acclimation at room temperature. A mathematical model was applied to describe E. coli O157 regrowth potential in compost. Repopulation occurred in autoclaved compost with a moisture content as low as 20% (water activity of 0.986) in the presence of background microflora of 2.3 to 3.9 log CFU/g. The population of all three E. coli O157 strains increased from ca. 1 to 4.85 log CFU/g in autoclaved compost, with the highest increase in the spinach-outbreak strain. However, E. coli O157 regrowth was suppressed by background microflora at ca. 6.5 log CFU/g. By eliminating acclimation at room temperature and increasing the inoculum level to ca. 3 log CFU/g, E. coli O157:H7 could regrow in the presence of high levels of background microflora. E. coli O157:H7 regrowth in the autoclaved compost collected from the field study was evident at all sampling days, with the population increase ranging from 3.49 to 6.54 log CFU/g. The fate of E. coli O157:H7 in compost was well described by a Whiting and Cygnarowicz-Provost model, with R2 greater than 0.9. The level of background microflora was a significant factor for both growth and death parameters. Our results reveal that a small number of E. coli O157 cells can regrow in compost, and both background microflora and moisture content were major factors affecting E. coli O157:H7 growth.

Validation of Apple Cider Pasteurization Treatments against Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes

2001 ◽  
Vol 64 (11) ◽  
pp. 1679-1689 ◽  
Author(s):  
PEGGY P. MAK ◽  
BARBARA H. INGHAM ◽  
STEVEN C. INGHAM
Keyword(s):  
Escherichia Coli ◽  
New York ◽  
Listeria Monocytogenes ◽  
Fluorescent Protein ◽  
Escherichia Coli O157 ◽  
Salmonella Spp ◽  
Apple Cider ◽  
E Coli ◽  
Log Reduction ◽  
Green Fluorescent

Time and temperature pasteurization conditions common in the Wisconsin cider industry were validated using a six-strain cocktail of Escherichia coli O157:H7 and acid-adapted E. coli O157:H7 in pH- and ∘Brix-adjusted apple cider. Strains employed were linked to outbreaks (ATCC 43894 and 43895, C7927, and USDA-FSIS-380–94) or strains engineered to contain the gene for green fluorescent protein (pGFP ATCC 43894 and pGFP ATCC 43889) for differential enumeration. Survival of Salmonella spp. (CDC 0778, CDC F2833, and CDC HO662) and Listeria monocytogenes (H0222, F8027, and F8369) was also evaluated. Inoculated cider of pH 3.3 or 4.1 and 11 or 14°Brix was heated under conditions ranging from 60°C for 14 s to 71.1°C for 14 s. A 5-log reduction of nonadapted and acid-adapted E. coli O157:H7 was obtained at 68.1°C for 14 s. Lower temperatures, or less time at 68.1°C, did not ensure a 5-log reduction in E. coli O157:H7. A 5-log reduction was obtained at 65.6°C for 14 s for Salmonella spp. L. monocytogenes survived 68.1°C for 14 s, but survivors died in cider within 24 h at 4°C. Laboratory results were validated with a surrogate E. coli using a bench-top plate heat-exchange pasteurizer. Results were further validated using fresh unpasteurized commercial ciders. Consumer acceptance of cider pasteurized at 68.1°C for 14 s (Wisconsin recommendations) and at 71.1°C for 6 s (New York recommendations) was not significantly different. Hence, we conclude that 68.1°C for 14 s is a validated treatment for ensuring adequate destruction of E. coli O157:H7, Salmonella spp., and L. monocytogenes in apple cider.

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