Escherichia coli O157: H7 Acid Tolerance and Survival in Apple Cider

1994 ◽  
Vol 57 (6) ◽  
pp. 460-464 ◽  
Author(s):  
LESLIE GARLAND MILLER ◽  
CHARLES W. KASPAR
Keyword(s):  
Escherichia Coli ◽  
Sodium Benzoate ◽  
Acid Tolerance ◽  
Potassium Sorbate ◽  
Escherichia Coli O157 ◽  
Control Strain ◽  
High Ph ◽  
Apple Cider ◽  
E Coli ◽  
Log Reduction

The survival of two Escherichia coli O157:H7 (ATCC 43889 and 43895) and a control strain E. coli was compared in apple cider and in Trypticase soy broth (TSB) adjusted to low and high pH. The O157:H7 strains were detectable in apple cider after 14 to 21 days at 4°C, whereas the control strain could not be detected (> 4-log reduction) after 5 to 7 days. During the first 14 days of storage at 4°C, the levels of strain 43889 decreased by ~3 logs, whereas levels of strain 43895 were unchanged. Survival of O157:H7 strains and the control strain were unaffected by the presence of potassium sorbate or sodium benzoate, except in one instance. Sodium benzoate caused a decrease of 57% in strain 43895 after 21 days, but ~104 CFU/ml still remained. In TSB adjusted to pH 2, 3, 4, 11 or 12, strain 43895 was again the more resistant of the O157:H7 strains, both of which were more durable than the control strain. The O157:H7 strains (especially strain 43895) withstood pH 2 with a minimal drop in CPU after 24 h, whereas no viable organisms were detectable after this time at pH 12. At these extremes of pH, survival was generally greater at 4°C than at 25°C. Despite differences between strains, these results show that E. coli O157:H7 is exceptionally tolerant of acid pH.

Heat Inactivation of Escherichia coli O157:H7 in Apple Cider Containing Malic Acid, Sodium Benzoate, and Potassium Sorbate

2000 ◽  
Vol 63 (8) ◽  
pp. 1026-1031 ◽  
Author(s):  
L. LOTTE DOCK ◽  
JOHN D. FLOROS ◽  
RICHARD H. LINTON
Keyword(s):  
Escherichia Coli ◽  
Malic Acid ◽  
Sodium Benzoate ◽  
Thermal Inactivation ◽  
Potassium Sorbate ◽  
Heat Inactivation ◽  
Escherichia Coli O157 ◽  
Additive Effects ◽  
Apple Cider ◽  
Log Reduction

The effect of pH modification and preservative addition in apple cider on the heat resistance of Escherichia coli O157: H7 was investigated. E. coli O157:H7 and various amounts of potassium sorbate (0 to 0.2%), sodium benzoate (0 to 0.2%), and malic acid (0 to 1%) were added to apple cider. Thermal inactivation experiments were performed at 47, 50, and 53°C, and D- and z-values were calculated. In apple cider without additives, the D-value at 50°C (D50) was about 65 min, but addition of preservatives and malic acid significantly (P < 0.01) decreased D-values. D50-values decreased to 13.9 min in cider with 0.5% malic acid, 13.2 min with 0.1% sorbate, and 7.0 min with 0.1% benzoate added. Addition of both sorbate and malic acid had similar effects as either one alone, thus additive effects were not present. However, addition of both 0.2% benzoate and 1% malic acid did show additive effects, lowering D50 to 0.3 min. Addition of all three components (0.2% sorbate, 0.2% benzoate, and 1% malic acid) resulted in a D50 = 18 s. The z-value of cider without additives was about 6°C, whereas z-values of cider containing malic acid, benzoate, and/or sorbate ranged from about 6°C to 26°C. This increase may result in a longer 5-log reduction time at higher temperatures (i.e., 70°C) in cider with benzoate as compared to cider without additives.

Addition of Fumaric Acid and Sodium Benzoate as an Alternative Method To Achieve a 5-log Reduction of Escherichia coli O157:H7 Populations in Apple Cider

2002 ◽  
Vol 65 (3) ◽  
pp. 476-483 ◽  
Author(s):  
JUSTIN E. COMES ◽  
ROBERT B. BEELMAN
Keyword(s):  
Escherichia Coli ◽  
Fumaric Acid ◽  
Sodium Benzoate ◽  
Bactericidal Effect ◽  
Escherichia Coli O157 ◽  
Refrigerated Storage ◽  
Apple Cider ◽  
E Coli ◽  
Log Reduction ◽  
Strong Linear Correlation

A study was conducted to develop a preservative treatment capable of the Food and Drug Administration–mandated 5-log reduction of Escherichia coli O157:H7 populations in apple cider. Unpreserved apple cider was treated with generally recognized as safe acidulants and preservatives before inoculation with E. coli O157:H7 in test tubes and subjected to mild heat treatments (25, 35, and 45°C) followed by refrigerated storage (4°C). Fumaric acid had significant (P < 0.05) bactericidal effect when added to cider at 0.10% (wt/vol) and adjusted to pH 3.3, but citric and malic acid had no effect. Strong linear correlation (R2 = 0.96) between increasing undissociated fumaric acid concentrations and increasing log reductions of E. coli O157:H7 in apple cider indicated the undissociated acid to be the bactericidal form. The treatment that achieved the 5-log reduction in three commercial ciders was the addition of fumaric acid (0.15%, wt/vol) and sodium benzoate (0.05%, wt/vol) followed by holding at 25°C for 6 h before 24 h of refrigeration at 4°C. Subsequent experiments revealed that the same preservatives added to cider in flasks resulted in a more than 5-log reduction in less than 5 and 2 h when held at 25 and 35°C, respectively. The treatment also significantly (P < 0.05) reduced total aerobic counts in commercial ciders to populations less than those of pasteurized and raw ciders from the same source (after 5 and 21 days of refrigerated storage at 4°C, respectively). Sensory evaluation of the same ciders revealed that consumers found the preservative-treated cider to be acceptable.

Efficacy of Ultraviolet Light for Reducing Escherichia coli O157:H7 in Unpasteurized Apple Cider

2000 ◽  
Vol 63 (5) ◽  
pp. 563-567 ◽  
Author(s):  
J. R. WRIGHT ◽  
S. S. SUMNER ◽  
C. R. HACKNEY ◽  
M. D. PIERSON ◽  
B. W. ZOECKLEIN
Keyword(s):  
Thin Film ◽  
Escherichia Coli ◽  
Uv Light ◽  
Uv Disinfection ◽  
Escherichia Coli O157 ◽  
Apple Cider ◽  
E Coli ◽  
Log Reduction ◽  
Disinfection Unit ◽  
Additional Reduction

This study examined the efficacy of UV light for reducing Escherichia coli O157:H7 in unpasteurized cider. Cider containing a mixture of acid-resistant E. coli O157:H7 (6.3 log CFU/ml) was treated using a thin-film UV disinfection unit at 254 nm. Dosages ranged from 9,402 to 61,005 μW-s/cm2. Treatment significantly reduced E. coli O157:H7 (P ≤ 0.0001). Mean reduction for all treated samples was 3.81 log CFU/ml. Reduction was also affected by the level of background microflora in cider. Results indicate that UV light is effective for reducing this pathogen in cider. However, with the dosages used in this experiment, additional reduction measures are necessary to achieve the required 5-log reduction.

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.

Antimicrobial Effects of Weak Acids on the Survival of Escherichia coli O157:H7 under Anaerobic Conditions†‡

2011 ◽  
Vol 74 (6) ◽  
pp. 893-898 ◽  
Author(s):  
HUIYING J. LU ◽  
FREDERICK BREIDT ◽  
ILENYS M. PÉREZ-DÍAZ ◽  
JASON A. OSBORNE
Keyword(s):  
Escherichia Coli ◽  
Food Products ◽  
Escherichia Coli O157 ◽  
Weak Acids ◽  
Anaerobic Conditions ◽  
Antimicrobial Effects ◽  
Apple Cider ◽  
E Coli ◽  
Log Reduction ◽  
Antimicrobial Effectiveness

Outbreaks of disease due to vegetative bacterial pathogens associated with acid foods (such as apple cider) have raised concerns about acidified vegetables and related products that have a similar pH (3.2 to 4.0). Escherichia coli O157:H7 and related strains of enterohemorrhagic E. coli (EHEC) have been identified as the most acid resistant vegetative pathogens in these products. Previous research has shown that the lack of dissolved oxygen in many hermetically sealed acid or acidified food products can enhance survival of EHEC compared with their survival under aerobic conditions. We compared the antimicrobial effects of several food acids (acetic, malic, lactic, fumaric, benzoic, and sorbic acids and sulfite) on a cocktail of EHEC strains under conditions representative of non–heat-processed acidified vegetables in hermetically sealed jars, holding the pH (3.2) and ionic strength (0.342) constant under anaerobic conditions. The overall antimicrobial effectiveness of weak acids used in this study was ranked, from most effective to least effective: sulfite > benzoic acid > sorbic acid > fumaric acid > l- and d-lactic acid > acetic acid > malic acid. These rankings were based on the estimated protonated concentrations required to achieve a 5-log reduction in EHEC after 24 h of incubation at 30°C. This study provides information that can be used to formulate safer acid and acidified food products and provides insights about the mode of action of weak acids against EHEC.

Effects of Cetylpyridinium Chloride, Acidified Sodium Chlorite, and Potassium Sorbate on Populations of Escherichia coli O157:H7, Listeria monocytogenes, and Staphylococcus aureus on Fresh Beef†

2004 ◽  
Vol 67 (2) ◽  
pp. 310-315 ◽  
Author(s):  
KYUNGWHA LIM ◽  
AZLIN MUSTAPHA
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Listeria Monocytogenes ◽  
Cetylpyridinium Chloride ◽  
Potassium Sorbate ◽  
Sodium Chlorite ◽  
Escherichia Coli O157 ◽  
Mixed Solution ◽  
E Coli ◽  
Log Reduction

The effects of selected food-grade antimicrobial agents at decreasing the number of pathogenic bacteria on fresh beef were determined. Beef cubes inoculated with Escherichia coli O157:H7, Listeria monocytogenes, or Staphylococcus aureus were sprayed with 0.5% cetylpyridinium chloride (CPC), 0.12% acidified sodium chlorite (ASC), 0.1% potassium sorbate (PS), or an equal mix of any two solutions. The beef samples were placed on absorbent tray pads sprayed with each single or mixed solution, wrapped with polyvinyl chloride film, heat sealed, and stored at 4°C for 2 weeks. Surface sanitization using CPC, ASC, or an equal mix of these two agents effectively reduced microbial numbers on the beef during storage. At day 0, ASC and the CPC-ASC mix reduced the number of E. coli O157:H7 by 2.50 and 1.58 log CFU/cm2, respectively. CPC demonstrated a 3.25-log reduction of L. monocytogenes and a 4.70-log reduction of S. aureus at 14 days. The CPC-PS mix reduced E. coli O157:H7 numbers by 1.46, L. monocytogenes by 2.95, and S. aureus by 4.41 log CFU/cm2 at 14 days. PS alone and the mixed solutions, CPC-ASC, CPC-PS, or ASC-PS, were not as effective as ASC or CPC alone. To effectively reduce E. coli O157:H7, L. monocytogenes, or S. aureus numbers, higher (>0.1%) concentrations of PS were necessary. Loss of redness and light color of beef surfaces consistently coincided with decreases in pH for ASC-treated beef samples.

PCR for the Specific Detection of an Escherichia coli O157:H7 Laboratory Control Strain

2015 ◽  
Vol 78 (9) ◽  
pp. 1738-1744 ◽  
Author(s):  
MICHAEL KNOWLES ◽  
DOMINIC LAMBERT ◽  
GEORGE HUSZCZYNSKI ◽  
MARTINE GAUTHIER ◽  
BURTON W. BLAIS
Keyword(s):  
Escherichia Coli ◽  
Pathogenic Bacteria ◽  
Control Measure ◽  
Positive Control ◽  
Food Microbiology ◽  
Escherichia Coli O157 ◽  
Control Strain ◽  
E Coli ◽  
Signature Sequences ◽  
Materials Used

Control strains of bacterial pathogens such as Escherichia coli O157:H7 are commonly processed in parallel with test samples in food microbiology laboratories as a quality control measure to assure the satisfactory performance of materials used in the analytical procedure. Before positive findings can be reported for risk management purposes, analysts must have a means of verifying that pathogenic bacteria (e.g., E. coli O157:H7) recovered from test samples are not due to inadvertent contamination with the control strain routinely handled in the laboratory environment. Here, we report on the application of an in-house bioinformatic pipeline for the identification of unique genomic signature sequences in the development of specific oligonucleotide primers enabling the identification of a common positive control strain, E. coli O157:H7 (ATCC 35150), using a simple PCR procedure.

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.

Viability of Acid-Adapted Escherichia coli O157:H7 in Ground Beef Treated with Acidic Calcium Sulfate

2004 ◽  
Vol 67 (3) ◽  
pp. 591-595 ◽  
Author(s):  
LARRY R. BEUCHAT ◽  
ALAN J. SCOUTEN
Keyword(s):  
Escherichia Coli ◽  
Acetic Acid ◽  
Calcium Sulfate ◽  
Ground Beef ◽  
Storage Period ◽  
Acid Tolerance ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Acidic Environments ◽  
Subsequent Acid

The effects of lactic acid, acetic acid, and acidic calcium sulfate (ACS) on viability and subsequent acid tolerance of three strains of Escherichia coli O157:H7 were determined. Differences in tolerance to acidic environments were observed among strains, but the level of tolerance was not affected by the acidulant to which cells had been exposed. Cells of E. coli O157:H7 adapted to grow on tryptic soy agar acidified to pH 4.5 with ACS were compared to cells grown at pH 7.2 in the absence of ACS for their ability to survive after inoculation into ground beef treated with ACS, as well as untreated beef. The number of ACS-adapted cells recovered from ACS-treated beef was significantly (α = 0.05) higher than the number of control cells recovered from ACS-treated beef during the first 3 days of a 10-day storage period at 4°C, suggesting that ACS-adapted cells might be initially more tolerant than unadapted cells to reduced pH in ACS-treated beef. Regardless of treatment of ground beef with ACS or adaptation of E. coli O157:H7 to ACS before inoculating ground beef, the pathogen survived in high numbers.

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