Enumeration of Escherichia coli O157:H7 in Outbreak-Associated Beef Patties

2016 ◽  
Vol 79 (7) ◽  
pp. 1266-1268 ◽  
Author(s):  
ALEXANDER GILL ◽  
GEORGE HUSZCZYNSKI

ABSTRACT An outbreak of five cases of Escherichia coli O157 infection that occurred in Canada in 2012 was linked to frozen beef patties seasoned with garlic and peppercorn. Unopened retail packs of beef patties from the implicated production lot were recovered and analyzed to enumerate E. coli O157, other E. coli strains, and total coliforms. E. coli O157 was not recovered by direct enumeration on selective agar media. E. coli O157 in the samples was estimated at 3.1 most probable number per 140 g of beef patty, other E. coli was 11 CFU/g, and coliforms were 120 CFU/g. These results indicate that the presence of E. coli O157 in ground beef at levels below 0.1 CFU/g may cause outbreaks. However, the roles of temperature abuse, undercooking, and cross-contamination in amplifying the risk are unknown.

1999 ◽  
Vol 122 (2) ◽  
pp. 185-192 ◽  
Author(s):  
J. TUTTLE ◽  
T. GOMEZ ◽  
M. P. DOYLE ◽  
J. G. WELLS ◽  
T. ZHAO ◽  
...  

Between November 1992 and February 1993, a large outbreak of Escherichia coli O157[ratio ]H7 infections occurred in the western USA and was associated with eating ground beef patties at restaurants of one fast-food chain. Restaurants that were epidemiologically linked with cases served patties produced on two consecutive dates; cultures of recalled ground beef patties produced on those dates yielded E. coli O157[ratio ]H7 strains indistinguishable from those isolated from patients, confirming the vehicle of illness. Seventy-six ground beef patty samples were cultured quantitatively for E. coli O157[ratio ]H7. The median most probable number of organisms was 1·5 per gram (range, <0·3–15) or 67·5 organisms per patty (range, <13·5–675). Correlation of the presence of E. coli O157[ratio ]H7 with other bacterial indicators yielded a significant association between coliform count and the presence of E. coli O157[ratio ]H7 (P=0·04). A meat traceback to investigate possible sources of contamination revealed cattle were probably initially colonized with E. coli O157[ratio ]H7, and that their slaughter caused surface contamination of meat, which once combined with meat from other sources, resulted in a large number of contaminated ground beef patties. Microbiological testing of meat from lots consumed by persons who became ill was suggestive of an infectious dose for E. coli O157[ratio ]H7 of fewer than 700 organisms. These findings present a strong argument for enforcing zero tolerance for this organism in processed food and for markedly decreasing contamination of raw ground beef. Process controls that incorporate microbiological testing of meat may assist these efforts.


2019 ◽  
Vol 2 (2) ◽  
pp. a13-19
Author(s):  
ELEXSON NILLIAN ◽  
AMIZA NUR ◽  
DIYANA NUR ◽  
AMIRAH ZAKIRAH ◽  
GRACE BEBEY

Contamination of drinks with E. coli O157:H7 served in food premises such as restaurants can cause haemorrhagic colitis and haemolytic uremic syndrome to humans. The presence or absence of faecal pathogen was demonstrated using coliform group as indicator microorganisms. Therefore, this study was conducted to detect the presence of E. coli O157:H7 in drinking water from food restaurant premise in Kota Samarahan and Kuching to ensure safe and potable drinking water is served to the consumer. A total of thirty (n=30) drink samples including six types of each of the samples are cold plain water, iced tea, iced milo, syrup and iced milk tea. Most Probable Number (MPN) procedure was used in this study to enumerate the MPN values of coliform bacteria in each drink collected. A total of 53.33% (16/30) of the drink samples showed positive E. coli detection. Then, the PCR assay showed 6.25% (one out of 16 isolates) samples were positive and carried stx1 gene produced by E. coli O157:H7 in iced milo sample types. This study showed the drinks collected from food premises was contaminated with faecal contamination, which was not safe to drink by the consumer. Therefore, preventive actions should be taken to prevent foodborne illness outbreak in future


2007 ◽  
Vol 70 (7) ◽  
pp. 1663-1669 ◽  
Author(s):  
PINA M. FRATAMICO ◽  
LORI K. BAGI

A comparison was made of the relative efficiencies of three enrichment media, RapidChek Escherichia coli O157:H7 enrichment broth (REB), R&F broth (RFB), and modified E. coli broth containing novobiocin (mEC+n), and four selective plating media for detection of cold- and freeze-stressed E. coli O157:H7 in raw ground beef. Ground beef (25 g) was inoculated with E. coli O157:H7 at ≤0.5 and ≤2 CFU/g, and samples were then enriched immediately or were stored at 4°C for 72 h or at −20°C for 2 weeks and then enriched. After 8 or 20 h of enrichment, the cultures were plated onto R&F E. coli O157: H7 chromogenic plating medium, cefixime-tellurite sorbitol MacConkey agar, CHROMagar O157, and Rainbow agar O157 and tested using the RapidChek E. coli O157 lateral flow immunoassay and a multiplex PCR assay targeting the E. coli O157: H7 eae, stx1, and stx2 genes. Recovery of E. coli O157:H7 on the four agar media was 4.0 to 7.9 log CFU/ml with the REB enrichment, 1.4 to 7.4 log CFU/ml with RFB, 1.7 to 6.7 log CFU/ml with mEC+n incubated at 42°C, and 1.3 to 3.3 log CFU/ml from mEC+n incubated at 35°C. The percentages of positive ground beef samples containing nonstressed, cold-stressed, and freeze-stressed E. coli O157:H7 as obtained by plating, the immunoassay, and the PCR assay were 97, 88, and 97%, respectively, with REB, 92, 81, and 78%, respectively, with RFB, 97, 58, and 53%, respectively, with mEC+n incubated at 42°C, and 22, 31, and 25%, respectively, with mEC+n incubated at 35°C. Logistic regression analyses of the data indicated significant main effects of treatment, type of medium, enrichment time, inoculum concentration, and detection method. In particular, a positive result was 1.1 times more likely to occur after 20 h of enrichment than after 8 h, 25 times more likely with RFB and REB than with mEC+nat35°C, 3.7 times more likely with an initial inoculum of ≤2.0 CFU/g than with ≤0.5 CFU/g, 2.5 to 3 times more likely using freeze-stressed or nonstressed bacteria than with cold-stressed bacteria, and 2.5 times more likely by plating than by the immunoassay or the PCR assay. REB had better overall performance for enrichment of cold- and freeze-stressed E. coli O157:H7 present in ground beef than did the other media examined.


1999 ◽  
Vol 62 (11) ◽  
pp. 1243-1247 ◽  
Author(s):  
SUSAN E. ANSAY ◽  
KIM A. DARLING ◽  
CHARLES W. KASPAR

The survival of Escherichia coli O157:H7 and of a nonpathogenic control strain of E. coli was monitored in raw ground beef that was stored at 2°C for 4 weeks, −2°C for 4 weeks, 15°C for 4 h and then −2°C for 4 weeks, and −20°C. Irradiated ground beef was inoculated with one E. coli control strain or with a four-strain cocktail of E. coli O157:H7 (ca. 105 CFU/g), formed into patties (30 to 45 g), and stored at the appropriate temperature. The numbers of the E. coli control strain decreased by 1.4 log10 CFU/g, and pathogen numbers declined 1.9 log10 CFU/g when patties were stored for 4 weeks at 2°C. When patties were stored at −2°C for 4 weeks, the numbers of the E. coli control strain and the serotype O157:H7 strains decreased 2.8 and 1.5 log10 CFU/g, respectively. Patties stored at 15°C for 4 h prior to storage at −2°C for 4 weeks resulted in 1.6 and 2.7 log10–CFU/g reduction in the numbers of E. coli and E. coli O157:H7, respectively. Storage of retail ground beef at 15°C for 4 h (tempering) did not result in increased numbers of colony forming units per gram, as determined with violet red bile, MRS lactobacilli, and plate-count agars. Frozen storage (−20°C) of ground-beef patties that had been inoculated with a single strain of E. coli resulted in approximately a 1 to 2 log10–CFU/g reduction in the numbers of the control strain and individual serotype O157:H7 strains after 1 year. There was no significant difference between the survival of the control strain and the O157:H7 strains, nor was there a difference between O157:H7 strains. These data demonstrate that tempering of ground-beef patties prior to low-temperature storage accelerated the decline in the numbers of E. coli O157:H7.


2008 ◽  
Vol 71 (4) ◽  
pp. 811-815 ◽  
Author(s):  
PILAR MORALES ◽  
JAVIER CALZADA ◽  
MARTA ÁVILA ◽  
MANUEL NUÑEZ

The effect of single- and multiple-cycle high-pressure treatments on the survival of Escherichia coli CECT 4972, a strain belonging to the O157:H7 serotype, in ground beef was investigated. Beef patties were inoculated with 107 CFU/g E. coli O157:H7, and held at 4°C for 20 h before high-pressure treatments. Reduction of the E. coli O157:H7 population by single-cycle treatments at 400 MPa and 12°C ranged from 0.82 log CFU/g for a 1-min cycle to 4.39 log CFU/g for a 20-min cycle. Multiple-cycle treatments were very effective, with four 1-min cycles at 400 MPa and 12°C reducing the E. coli O157:H7 population by 4.38 log CFU/g, and three 5-min cycles by 4.96 log CFU/g. The color parameter L* increased significantly with high-pressure treatments in the interior and the exterior of beef patties, whereas a* decreased in the interior, and b* increased in the exterior—changes that might diminish consumer acceptance of the product. Kramer shear force and energy were generally higher in pressurized than in control ground beef. Maximum values for these texture parameters, which corresponded to tougher patties, were reached after one 10-min cycle in the case of single-cycle treatments or two 5-min cycles in the case of multiple-cycle treatments. High-pressure treatments had no significant effect on Warner-Bratzler shear force.


1996 ◽  
Vol 59 (3) ◽  
pp. 230-237 ◽  
Author(s):  
TIMOTHY C. JACKSON ◽  
MARGARET D. HARDIN ◽  
GARY R. ACUFF

Stationary-phase cultures of Escherichia coli O157:H7 were inoculated into tryptic soy broth, sealed in vials, and stored at −18°C for 1, 8, and 15 days, or 3 or 15°C for 3, 6, and 9 h. Thermal resistance was determined at 55°C. Each storage treatment was repeated with additional holding at 23 or 30°C for 1, 2, 3, or 4 h prior to heating to simulate potential temperature abuse during handling. Cultures under treatments enabling the growth of E. coli O157:H7 were generally more heat sensitive than those held at temperatures which restricted growth or enabled growth to stationary phase. Cultures stored frozen (−18°C) without holding at elevated temperatures had greater heat resistance than those stored under refrigeration (3°C) or at 15°C. Subsequent holding of frozen cultures at 23 or 30°C resulted in a decrease in heat resistance. To determine whether these responses would be observed under typical commercial preparation procedures, ground beef patties were inoculated with E. coli O157:H7 and stored at 3 or 15°C for 9 h or at −18°C for 8 d and then held at 21 or 30°C for 0 or 4 h. Patties were grilled to an internal temperature of 54.4°C (130°F), 62.8°C (145°F), or 68.3°C (155°F). Cultures were most resistant in frozen patties, while cultures in patties stored at 15°C were the most heat sensitive. Holding patties at 21 or 30°C prior to grilling resulted in increased sensitivity. Storage and holding temperatures similar to those encountered in food service may influence the ability of E. coli O157:H7 to survive heat treatments.


2005 ◽  
Vol 68 (3) ◽  
pp. 451-457 ◽  
Author(s):  
NARELLE FEGAN ◽  
GLEN HIGGS ◽  
PAUL VANDERLINDE ◽  
PATRICIA DESMARCHELIER

The extent of contamination with Escherichia coli O157 was determined for 100 cattle during slaughter. Samples from 25 consecutively slaughtered cattle from four unrelated groups were collected from the oral cavity, hide, rumen, feces after evisceration, and pre- and postchill carcass. Ten random fecal samples were collected from the pen where each group of animals was held at the abattoir. E. coli O157 was detected using automated immunomagnetic separation (AIMS), and cell counts were determined using a combination of most probable number (MPN) and AIMS. E. coli O157 was isolated from 87 (14%) of the 606 samples collected, including 24% of 99 oral cavity samples, 44% of 100 hides, 10% of 68 fecal samples collected postevisceration, 6% of 100 prechill carcass swabs, and 15% of 40 fecal samples collected from holding pens. E. coli O157 was not isolated from rumen or postchill carcass samples. E. coli O157 was isolated from at least one sample from each group of cattle tested, and the prevalence in different groups ranged from less than 1 to 41%. The numbers of E. coli O157 differed among the animals groups. The group which contained the highest fecal (7.5 × 105 MPN/g) and hide (22 MPN/cm2) counts in any individual animal was the only group in which E. coli O157 was isolated from carcasses, suggesting a link between the numbers of E. coli O157 present and the risk of carcass contamination. Processing practices at this abattoir were adequate for minimizing contamination of carcasses, even when animals were heavily contaminated with E. coli O157.


1987 ◽  
Vol 70 (6) ◽  
pp. 991-993
Author(s):  
Paul L Poelma ◽  
Clyde R Wilson ◽  
Wallace H Andrews

Abstract An assay for the enzyme glucuronidase was used to determine the presence of Escherichia coli in selected, naturally contaminated high moisture foods. Raw pork sausage, ground turkey, and ground beef were inoculated into tubes containing the substrate 4-methylumbelliferyl beta-D-glucuronide (MUG) in lauryl tryptose (LT) medium. After incubation at 35°C for 24 h, the inoculated LT-MUG tubes were examined under longwave ultraviolet light for the presence of a fluorogenic glucuronidase end product. A fluorescing tube indicated the presumptive presence of E. coli. The 10 day most probable number method of the AOAC and the LT-MUG procedure gave comparable recoveries of E. coli.


2015 ◽  
Vol 78 (9) ◽  
pp. 1733-1737 ◽  
Author(s):  
ALEXANDER GILL ◽  
DENISE OUDIT

In this article, we discuss the enumerative analysis for Escherichia coli O157 in two raw milk Gouda cheese products (A and B), implicated in an outbreak of 29 cases of E. coli O157:H7 illness that occurred across Canada in 2013. Samples were enumerated for E. coli O157 by most probable number (MPN) over a period of 30 to 60 days after the end of the outbreak. Samples (55.55 g) of product A (n = 14) were analyzed at 146 to 180 days postproduction. E. coli O157 was isolated from six samples at 19.9 to 44.6 MPN/kg. The E. coli O157 concentration of product A estimated from the results of all 14 samples was 9.5 MPN/kg. Samples (55.55 g) of product B (n = 20) were analyzed at 133 to 149 days postproduction. E. coli O157 was isolated from four samples at 19.9 MPN/kg. The E. coli O157 concentration of product B estimated from the results of all 20 samples was 3.7 MPN/kg. Analysis of a 305-g sample of product A (n = 1) stored at 4°C until 306 days postproduction revealed that the E. coli O157 concentration had declined to 3.6 MPN/kg. E. coli O157 could not be isolated from 555-g samples of product B (n = 5) after 280 days postproduction. The physicochemical parameters (pH, water activity, percent moisture, and percent salt) of both cheese products were found to be in the normal range for this type of product. The results of this study demonstrate that E. coli O157 could not replicate during storage at 4°C in the products tested but was capable of survival following aging and prolonged storage. This indicates that, if contaminated, the minimum 60-day aging period, which is required for raw milk Gouda cheeses, is not sufficient in all cases to ensure that the product does not contain viable cells of E. coli O157. The results also indicate that samples sizes greater than 100 g may be required to reliably detect E. coli O157 in cheese products associated with outbreaks.


2015 ◽  
Vol 78 (8) ◽  
pp. 1547-1553 ◽  
Author(s):  
GORDON R. DAVIDSON ◽  
JOHN C. FRELKA ◽  
MAI YANG ◽  
THOMAS M. JONES ◽  
LINDA J. HARRIS

Inshell walnuts collected from California walnut handlers over four harvests were evaluated for the presence of Escherichia coli O157:H7 and Salmonella. E. coli O157:H7 was not detected in any of 2,903 375-g samples evaluated in 2011, 2012, and 2013 (&lt;0.034% prevalence; 95% confidence interval [CI], 0 to 0.13%). Salmonella was not isolated from any of the 935 samples in 2010 (100 g evaluated; &lt;0.11% prevalence; 95% CI, 0 to 0.41%) but was isolated from 2 of 905 (375 g; 0.22% prevalence; 95% CI, 0.061 to 0.80%), 1 of 998 (375 g; 0.10% prevalence; 95% CI, 0.018 to 0.56%), and 1 of 1,000 (375 g; 0.10% prevalence; 95% CI, 0.018 to 0.56%) samples in 2011, 2012, and 2013, respectively, for an average annual prevalence of 0.14% (375 g; 95% CI, 0.054 to 0.35%). The levels of Salmonella in positive samples determined by a modified most-probable-number (MPN) method were estimated to be 0.32 to 0.42 MPN/100 g (95% CI, 0.045 to 3.6 MPN/100 g).


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