Behavior of Escherichia coli O157:H7 during the Manufacture and Aging of Gouda and Stirred-Curd Cheddar Cheeses Manufactured from Raw Milk

2010 ◽  
Vol 73 (12) ◽  
pp. 2217-2224 ◽  
Author(s):  
DENNIS J. D'AMICO ◽  
MARC J. DRUART ◽  
CATHERINE W. DONNELLY
Keyword(s):  
Escherichia Coli ◽  
Population Level ◽  
Raw Milk ◽  
Escherichia Coli O157 ◽  
Milk Whey ◽  
Selective Enrichment ◽  
Point Counts ◽  
E Coli ◽  
Unpasteurized Milk ◽  
Low Levels

This study was conducted to examine the fate of Escherichia coli O157:H7 during the manufacture and aging of Gouda and stirred-curd Cheddar cheeses made from raw milk. Cheeses were manufactured from unpasteurized milk experimentally contaminated with one of three strains of E. coli O157:H7 at an approximate population level of 20 CFU/ml. Samples of milk, whey, curd, and cheese were collected for enumeration of bacteria throughout the manufacturing and aging process. Overall, bacterial counts in both cheese types increased almost 10-fold from initial inoculation levels in milk to approximately 145 CFU/g found in cheeses on day 1. From this point, counts dropped significantly over 60 days to mean levels of 25 and 5 CFU/g in Cheddar and Gouda, respectively. Levels of E. coli O157:H7 fell and stayed below 5 CFU/g after an average of 94 and 108 days in Gouda and Cheddar, respectively, yet remained detectable after selective enrichment for more than 270 days in both cheese types. Changes in pathogen levels observed throughout manufacture and aging did not significantly differ by cheese type. In agreement with results of previous studies, our results suggest that the 60-day aging requirement alone is insufficient to completely eliminate levels of viable E. coli O157:H7 in Gouda or stirred-curd Cheddar cheese manufactured from raw milk contaminated with low levels of this pathogen.

scholarly journals Prevalence and molecular characterisation of shiga-toxin-producing Escherichia coli in raw milk cheeses from Lazio region, Italy

2016 ◽  
Vol 5 (1) ◽  
Author(s):  
Selene Marozzi ◽  
Paola De Santis ◽  
Sarah Lovari ◽  
Roberto Condoleo ◽  
Stefano Bilei ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Raw Milk ◽  
Foodborne Diseases ◽  
Control Plan ◽  
Lazio Region ◽  
Standard Methods ◽  
E Coli ◽  
Regional Control ◽  
Unpasteurized Milk

In recent years, the incidence of foodborne diseases caused by shiga toxin-producing <em>Escherichia coli</em> (STEC) has increased globally. For this reason, within the specific regional control plan for the detection of STEC in food products in Italy, the presence of STEC in unpasteurized milk cheeses was investigated. In total 203 samples obtained from March 2011 to December 2013 were analysed, with two standard methods (ISO 16654:2001 and ISO 13136:2012). Two strains of <em>E. coli</em> O157 were isolated (2/161, 1.2%) but did not carry any virulence-associated genes and 22 <em>stx</em>-positive samples (22/146, 15.1%) were detected in enrichment cultures, mostly from ovine cheeses. Only two strains isolated from different ovine cheeses carried <em>stx</em> gene and none of these was <em>eae</em>-positive. This study confirms the presence of <em>stx</em>-positive <em>E. coli</em> and suggests that this type of food cannot be excluded as a potential vehicle of STEC.

Horizontal Transmission of Escherichia coli O157:H7 during Cattle Housing

2004 ◽  
Vol 67 (12) ◽  
pp. 2651-2656 ◽  
Author(s):  
P. McGEE ◽  
L. SCOTT ◽  
J. J. SHERIDAN ◽  
B. EARLEY ◽  
N. LEONARD
Keyword(s):  
Escherichia Coli ◽  
Drinking Water ◽  
Animal Behavior ◽  
Horizontal Transmission ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Holstein Friesian ◽  
Water Feed ◽  
Low Levels ◽  
The Right

Ruminant livestock, particularly cattle, is considered the primary reservoir of Escherichia coli O157:H7. This study examines the transmission of E. coli O157:H7 within groups of cattle during winter housing. Holstein Friesian steers were grouped in six pens of five animals. An animal inoculated with and proven to be shedding a marked strain of E. coli O157: H7 was introduced into each pen. Fecal (rectal swabs) and hide samples (900 cm2 from the right rump) were taken from the 36 animals throughout the study. Water, feed, and gate or partition samples from each pen were also examined. Within 24 h of introducing the inoculated animals into the pens, samples collected from the drinking water, pen barriers, and animal hides were positive for the pathogen. Within 48 h, the hides of 20 (66%) of 30 cohort animals from the six pens were contaminated with E. coli O157:H7. The first positive fecal samples from the noninoculated cohort animals were detected 3 days after the introduction of the inoculated steers. During the 23 days of the study, 15 of 30 cohort animals shed the marked E. coli O157: H7 strain in their feces on at least one occasion. Animal behavior in the pens was monitored during a 12-h period using closed circuit television cameras. The camera footage showed an average of 13 instances of animal grooming in each pen per hour. The study suggests that transmission of E. coli O157:H7 between animals may occur following ingestion of the pathogen at low levels and that animal hide may be an important source of transmission.

Distribution Patterns of Escherichia coli O157:H7 in Ground Beef Produced by a Laboratory-Scale Grinder†

2002 ◽  
Vol 65 (12) ◽  
pp. 1894-1902 ◽  
Author(s):  
ROLANDO A. FLORES ◽  
MARK L. TAMPLIN
Keyword(s):  
Escherichia Coli ◽  
Ground Beef ◽  
Distribution Patterns ◽  
Small Scale ◽  
Escherichia Coli O157 ◽  
Inoculum Level ◽  
E Coli ◽  
Specific Distribution ◽  
Low Levels ◽  
G Prior

This study determined the distribution patterns of Escherichia coli O157:H7 in ground beef when a contaminated beef trim was introduced into a batch of uncontaminated beef trims prior to grinding in a small-scale laboratory grinder. A beef trim (15.3 ± 2 g) was inoculated with a rifampicin-resistant strain of E. coli O157:H7 (E. coli O157:H7rif) and introduced into a stream of noncontaminated beef (322 ± 33 g) prior to grinding. Seven inoculum levels (6, 5, and 4 total log CFU [high]; and 3, 2, 1, and 0 total log CFU [low]) were studied in triplicate. E. coli O157:H7rif was not detected in 3.1 to 43% of the ground beef inoculated with the high levels or in 3.4 to 96.9% of the ground beef inoculated with the low levels. For all inoculum levels studied, the five ground beef fractions (each 7.8 ± 0.6 g) with the highest pathogen levels accounted for 59 to 100% of the total pathogens detected. For all inoculum levels, there was a linear relationship between the quantity of ground beef containing E. coli O157:H7rif and the inoculum level. The quantity of E. coli O157:H7rif in the beef remaining in the grinder was proportional to the inoculum level and was related to the location in the grinder. Different components of the grinder accumulated E. coli O157:H7rif in different quantities, with the most significant accumulation being in the nut (collar) that attaches the die to the blade. This study determined specific distribution patterns of E. coli O157:H7rif after the grinding of a contaminated beef trim along with uncontaminated trims, and the results indicate that the grinding operation should be regarded as a means of distribution of microbial contamination in risk analyses of ground beef operations.

scholarly journals First isolation of Escherichia coli O157:H7 from faecal and milk specimens from Anatolian water buffaloes (Bubalus bubalus) in Turkey

2008 ◽  
Vol 79 (4) ◽  
Author(s):  
E. Seker ◽  
H. Yardimci
Keyword(s):  
Escherichia Coli ◽  
Raw Milk ◽  
Escherichia Coli O157 ◽  
Water Buffaloes ◽  
E Coli ◽  
Milk Samples ◽  
Cultural Methods ◽  
Faecal Samples

Three hundred rectal faecal samples and 213 raw milk samples obtained from the tanks and containers were examined using standard cultural methods. Escherichia coli O157:H7 was isolated from 11 (3.7 %) of 300 faecal samples and 3 (1.4 %) of 213 raw milk samples. It was determined that 8 (73 %) of E. coli O157:H7 strains isolated from faecal samples originated from water buffaloes younger than 2 years of age and 3 (27 %) from 2-year-old and older water buffaloes. This is the 1st isolation of Escherichia coli O157:H7 from faecal and milk samples of water buffaloes in Turkey.

scholarly journals Escherichia coli O157[ratio ]H7 diarrhoea associated with well water and infected cattle on an Ontario farm

1998 ◽  
Vol 120 (1) ◽  
pp. 17-20 ◽  
Author(s):  
S. G. JACKSON ◽  
R. B. GOODBRAND ◽  
R. P. JOHNSON ◽  
V. G. ODORICO ◽  
D. ALVES ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Phage Type ◽  
Dairy Farm ◽  
Female Child ◽  
Well Water ◽  
Escherichia Coli O157 ◽  
Cattle Farm ◽  
E Coli ◽  
Unpasteurized Milk ◽  
Hydrogeological Investigation

A 16-month old female child living on an Ontario dairy farm was taken to hospital suffering from bloody diarrhoea. Escherichia coli O157[ratio ]H7 was isolated from her stool. Initial tests of well water samples were negative for E. coli by standard methods but culture of selected coliform colonies on sorbitol-MacConkey agar led to isolation of E. coli O157[ratio ]H7. E. coli O157[ratio ]H7 was also isolated from 63% of cattle on the farm. The E. coli O157[ratio ]H7 isolates from the child, the water and the cattle were phage type 14, produced verotoxins 1 and 2, and were highly related on analysis by pulsed field gel electrophoresis. The child did not have known direct contact with the cattle and did not consume unpasteurized milk. Hydrogeological investigation revealed the design and location of the well would allow manure-contaminated surface water to flow into the well. This investigation demonstrates that cattle farm well water is a potential source of E. coli O157[ratio ]H7 which may not be identified by standard screening for E. coli in water.

Recirculating Immunomagnetic Separation and Optimal Enrichment Conditions for Enhanced Detection and Recovery of Low Levels of Escherichia coli O157:H7 from Fresh Leafy Produce and Surface Water

2007 ◽  
Vol 70 (12) ◽  
pp. 2717-2724 ◽  
Author(s):  
SUNEE HIMATHONGKHAM ◽  
MARY LEE DODD ◽  
JENNY K. YEE ◽  
DAVID K. LAU ◽  
RAYMOND G. BRYANT ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Surface Water ◽  
Real Time ◽  
Real Time Pcr ◽  
Immunomagnetic Separation ◽  
Escherichia Coli O157 ◽  
Simple Method ◽  
E Coli ◽  
Low Levels ◽  
Spinach Leaves

The objective of this study was to develop a rapid, simple method for enhanced detection and isolation of low levels of Escherichia coli O157:H7 from leafy produce and surface water using recirculating immunomagnetic separation (RIMS) coupled with real-time PCR and a standard culture method. The optimal enrichment conditions for the method also were determined. Analysis of real-time PCR data (CT values) suggested that incubation of lettuce and spinach leaves rather than rinsates provides better enrichment of E. coli O157:H7. Enrichment of lettuce or spinach leaves at 42°C for 5 h provided better detection than enrichment at 37°C. Extended incubation of surface water for 20 h at 42°C did not improve the detection. The optimized enrichment conditions were also employed with modified Moore swabs, which were used to sample flowing water sites. Positive isolation rates and real-time PCR results indicated an increased recovery of E. coli O157:H7 from all samples following the application of RIMS. Under these conditions, the method provided detection and/or isolation of E. coli O157:H7 at levels as low as 0.07 CFU/g of lettuce, 0.1 CFU/g of spinach, 6 CFU/100 ml of surface water, and 9 CFU per modified Moore swab. During a 6-month field study, modified Moore swabs yielded high isolation rates when deployed in natural watershed sites. The method used in this study was effective for monitoring E. coli O157:H7 in the farm environment, during postharvest processing, and in foodborne outbreak investigations.

Validation of Pepperoni Processes for Control of Escherichia coli O157:H7‡

1996 ◽  
Vol 59 (12) ◽  
pp. 1260-1266 ◽  
Author(s):  
JAY C. HINKENS ◽  
NANCY G. FAITH ◽  
TIMOTHY D. LORANG ◽  
PHILLIP BAILEY ◽  
DENNIS BUEGE ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Relative Humidity ◽  
Thermal Processing ◽  
Starter Culture ◽  
Escherichia Coli O157 ◽  
Protein Ratio ◽  
E Coli ◽  
Modified Method ◽  
Low Levels ◽  
Unit Reduction

The outbreak of Escherichia coli O157:H7 linked with dry-cured salami in late 1994 prompted regulatory action that required manufacturers of fermented products to demonstrate a 5-log unit reduction in counts of this pathogen during processing. Therefore, pepperoni batter (75% pork:25% beef with a fat content of ca. 32%) was inoculated with a pediococcal starter culture and a five-strain mixture of E. coli O157:H7 (≥2 × 107 CFU/g) and stuffed into 55-mm diameter fibrous casings 47 cm in length. The viability of the pathogen was monitored before stuffing, after fermentation, after thermal processing, and/or after drying. Chubs were fermented at 96°F (36°C) and 85% relative humidity (RH) to pH ≤ 5.0 and then dried at 55°F (13°C) and 65% RH to a moisture/protein ratio of ≤1.6:1 (modified method 6 process). Counts of the pathogen decreased about 1.2 log units after fermentation and drying. In subsequent experiments, heating chubs after fermentation to internal temperatures of 145°F (63°C) instantaneous or 128°F (53°C) for 60 min resulted in a ≥5-log unit decrease in numbers of strain O157:H7 without visibly affecting the texture or appearance of the product. These data revealed that a traditional nonthermal, process for pepperoni was only sufficient to eliminate relatively low levels (ca. 2 log CFU/g) of E. coli O157:H7, whereas heating to internal temperatures of 145°F (63°C) instantaneous or 128°F (53°C) for 60 min delivered a 5 to 6 log unit reduction in counts of the pathogen in pepperoni.

scholarly journals Contribution and Interaction of Shiga Toxin Genes to Escherichia coli O157:H7 Virulence

Toxins ◽  
2019 ◽  
Vol 11 (10) ◽  
pp. 607 ◽  
Author(s):  
Gillian A.M. Tarr ◽  
Taryn Stokowski ◽  
Smriti Shringi ◽  
Phillip I. Tarr ◽  
Stephen B. Freedman ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Population Level ◽  
Escherichia Coli O157 ◽  
Additive Interaction ◽  
Shiga Toxins ◽  
E Coli ◽  
Uremic Syndrome ◽  
Shiga Toxin Genes

Escherichia coli O157:H7 is the predominant cause of diarrhea-associated hemolytic uremic syndrome (HUS) worldwide. Its cardinal virulence traits are Shiga toxins, which are encoded by stx genes, the most common of which are stx1a, stx2a, and stx2c. The toxins these genes encode differ in their in vitro and experimental phenotypes, but the human population-level impact of these differences is poorly understood. Using Shiga toxin-encoding bacteriophage insertion typing and real-time polymerase chain reaction, we genotyped isolates from 936 E. coli O157:H7 cases and verified HUS status via chart review. We compared the HUS risk between isolates with stx2a and those with stx2a and another gene and estimated additive interaction of the stx genes. Adjusted for age and symptoms, the HUS incidence of E. coli O157:H7 containing stx2a alone was 4.4% greater (95% confidence interval (CI) −0.3%, 9.1%) than when it occurred with stx1a. When stx1a and stx2a occur together, the risk of HUS was 27.1% lower (95% CI −87.8%, −2.3%) than would be expected if interaction were not present. At the population level, temporal or geographic shifts toward these genotypes should be monitored, and stx genotype may be an important consideration in clinically predicting HUS among E. coli O157:H7 cases.

Transmission of Escherichia coli O157:H7 to Internal Tissues and Its Survival on Flowering Heads of Wheat

2015 ◽  
Vol 78 (3) ◽  
pp. 518-524 ◽  
Author(s):  
BISMARCK MARTINEZ ◽  
JAYNE STRATTON ◽  
ANDRÉIA BIANCHINI ◽  
STEPHEN WEGULO ◽  
GLEN WEAVER
Keyword(s):  
Escherichia Coli ◽  
Irrigation Water ◽  
Escherichia Coli O157 ◽  
Human Pathogen ◽  
Contamination Source ◽  
Wheat Seedlings ◽  
E Coli ◽  
Uremic Syndrome ◽  
Low Levels ◽  
Fourth Experiment

Escherichia coli O157:H7 is a human pathogen that can cause bloody diarrhea, hemorrhagic colitis, and hemolytic uremic syndrome. E. coli O157:H7 illnesses are mainly associated with undercooked beef; however, in recent years, outbreaks have been linked to fresh produce, such as spinach, lettuce, and sprouts. In 2009, flour was implicated as the contamination source in an outbreak involving consumption of raw cookie dough that resulted in 77 illnesses. The objectives of this research were to determine (i) whether E. coli O157:H7 could be translocated into the internal tissues of wheat (Triticum aestivum) seedlings from contaminated seed, soil, or irrigation water and (ii) whether the bacterium could survive on flowering wheat heads. The levels of contamination of kanamycin-resistant E. coli O157:H7 strains in seed, soil, and irrigation water were 6.88 log CFU/g, 6.60 log CFU/g, and 6.76 log CFU/ml, respectively. One hundred plants per treatment were sown in pot trays with 50 g of autoclaved soil or purposely contaminated soil, watered every day with 5 ml of water, and harvested 9 days postinoculation. In a fourth experiment, flowering wheat heads were spray inoculated with water containing 4.19 log CFU/ml E. coli O157:H7 and analyzed for survival after 15 days, near the harvest period. To detect low levels of internalization, enrichment procedures were performed and Biotecon real-time PCR detection assays were used to determine the presence of E. coli O157:H7 in the wheat, using a Roche Applied Science LightCycler 2.0 instrument. The results showed that internalization was possible using contaminated seed, soil, and irrigation water in wheat seedlings, with internalization rates of 2, 5, and 10%, respectively. Even though the rates were low, to our knowledge this is the first study to demonstrate the ability of this strain to reach the phylloplane in wheat. In the head contamination experiment, all samples tested positive, showing the ability of E. coli O157:H7 to survive on the wheat head.

Development of a Rapid Immunochromatographic Strip for Detection of Escherichia coli O157

2005 ◽  
Vol 68 (10) ◽  
pp. 2140-2143 ◽  
Author(s):  
BYEONG YEAL JUNG ◽  
SUK CHAN JUNG ◽  
CHANG HEE KWEON
Keyword(s):  
Escherichia Coli ◽  
High Specificity ◽  
Escherichia Coli O157 ◽  
Test Results ◽  
Positive Signal ◽  
Immunochromatographic Strip ◽  
Selective Enrichment ◽  
E Coli ◽  
Specificity And Sensitivity ◽  
Strip Test

We developed an immunochromatographic (IC) strip for the rapid detection of Escherichia coli O157 in enriched samples. Murine monoclonal antibody to E. coli O157:H7 lipopolysaccharide was conjugated with 40 nm of colloidal gold particles by the citrate method. The specificity of the IC strip was determined using 48 pure-cultured bacteria, including 32 E. coli strains and 16 non–E. coli strains. Regardless of H serotype, E. coli O157 strains produced a positive signal, whereas the others, representing 29 E. coli serotypes, did not. Among 16 non–E. coli strains, only Citrobacter amalonaticus yielded a positive signal. The sensitivity of the IC strip was determined using 10-fold diluted E. coli O157:H7, with a range of 1.8 × 107 to 1.8 CFU/ml in enriched raw beef. E. coli O157 could be detected at a minimum of 1.8 × 105 CFU/ml without enrichment and 1.8 CFU/ml after enrichment. Various samples were enriched to detect E. coli O157 using the IC strip and to isolate E. coli O157:H7 using traditional culture procedures. The IC strip test results exhibited 100% agreement with traditional methods after selective enrichment, since E. coli O157:H7 was also isolated from all the samples with positive strip test results. However, the specificity of the strip was somewhat higher with pork (98.8%) than with bovine feces (87.9%) and swine feces (93.4%). These results indicated that the IC strip exhibits high specificity and sensitivity in the detection of E. coli O157, and this assay is rapid, economical, and simple, without requirement of complicated equipment.

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