Rapid and Sensitive Detection of Escherichia coli O157:H7 in Milk and Ground Beef Using Magnetic Bead–Based Immunoassay Coupled with Tyramide Signal Amplification

2014 ◽  
Vol 77 (1) ◽  
pp. 100-105 ◽  
Author(s):  
MUHSIN AYDIN ◽  
GENE P. D. HERZIG ◽  
KWANG CHEOL JEONG ◽  
SAMANTHA DUNIGAN ◽  
PARTH SHAH ◽  
...  
Keyword(s):  
Public Health ◽  
Escherichia Coli ◽  
Food Safety ◽  
Foodborne Pathogens ◽  
Signal Amplification ◽  
Magnetic Bead ◽  
Escherichia Coli O157 ◽  
Tyramide Signal Amplification ◽  
E Coli ◽  
Enrichment Step

Escherichia coli O157:H7 is a major foodborne pathogen that has posed serious problems for food safety and public health. Recent outbreaks and recalls associated with various foods contaminated by E. coli O157:H7 clearly indicate its deleterious effect on food safety. A rapid and sensitive detection assay is needed for this harmful organism to prevent foodborne illnesses and control outbreaks in a timely manner. We developed a magnetic bead–based immunoassay for detection of E. coli O157:H7 (the most well-known Shiga toxigenic E. coli strain) with a 96-well microplate as an assay platform. Immunomagnetic separation (IMS) and tyramide signal amplification were coupled to the assay to increase its sensitivity and specificity. This immunoassay was able to detect E. coli O157:H7 in pure culture with a detection limit of 50 CFU/ml in less than 3 h without an enrichment step. The detection limit was decreased 10-fold to 5 CFU/ml with addition of a 3-h enrichment step. When this assay was tested with other nontarget foodborne pathogens and common enteric bacteria, no cross-reactivity was found. When tested with artificially contaminated ground beef and milk samples, the assay sensitivity decreased two- to fivefold, with detection limits of 250 and 100 CFU/ml, respectively, probably because of the food matrix effect. The assay results also were compared with those of a sandwich-type enzyme-linked immunosorbent assay (ELISA) and an ELISA coupled with IMS; the developed assay was 25 times and 4 times more sensitive than the standard ELISA and the IMS-ELISA, respectively. Tyramide signal amplification combined with IMS can improve sensitivity and specificity for detection of E. coli O157:H7. The developed assay could be easily adapted for other foodborne pathogens and will contribute to improved food safety and public health.

Evaluation of Molecular Typing Methods for Escherichia coli O157:H7 Isolates from Cattle, Food, and Humans

2004 ◽  
Vol 67 (4) ◽  
pp. 651-657 ◽  
Author(s):  
STEVEN L. FOLEY ◽  
SHABBIR SIMJEE ◽  
JIANGHONG MENG ◽  
DAVID G. WHITE ◽  
PATRICK F. McDERMOTT ◽  
...  
Keyword(s):  
Public Health ◽  
Escherichia Coli ◽  
Foodborne Pathogens ◽  
Molecular Typing ◽  
Housekeeping Gene ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Source Of Infection ◽  
Life Threatening ◽  
Typing Methods

Escherichia coli O157:H7, a Shiga toxin–producing E. coli, has been the causative agent of many cases of severe, often life-threatening foodborne illness. Because of the importance of E. coli O157:H7 to public health, many molecular typing methods have been developed to determine its transmission routes and source of infection during epidemiological investigations. Pulsed-field gel electrophoresis (PFGE) is currently used by public health organizations to track infections of E. coli O157:H7 and other foodborne pathogens. In this study, we compared the ability of PFGE, multilocus sequence typing (MLST), and repetitive-element PCR (Rep-PCR) to distinguish among 92 E. coli O157:H7 isolates from cattle, food, and infected humans. Several virulence genes, including the intimin gene (eaeA), the hemolysin gene (hlyA), and the H7 fimbrial gene (fliC), and a housekeeping gene for β-glucuronidase (uidA) were included in MLST. Rep-PCR reactions were performed using a commercially available typing kit (Bacterial Barcodes Inc., Houston, Tex.) with the provided Uprime-RI primer set. Results of the study indicated that PFGE provided the most discrimination among the techniques, identifying 72 distinct PFGE profiles for the isolates; Rep-PCR elucidated 14 different profiles, whereas MLST generated five profiles. Additionally, there did not appear to be any correlation among the typing methods examined in this study. Therefore, to date, PFGE remains the technique of choice for molecular subtyping of E. coli O157:H7.

Effect of Sampling Plans on the Risk of Escherichia coli O157 Illness

2015 ◽  
Vol 78 (7) ◽  
pp. 1370-1374
Author(s):  
ANDREAS KIERMEIER ◽  
JOHN SUMNER ◽  
IAN JENSON
Keyword(s):  
Public Health ◽  
Escherichia Coli ◽  
Sample Size ◽  
Health Effect ◽  
The United States ◽  
Escherichia Coli O157 ◽  
Sampling Protocol ◽  
Sampling Plans ◽  
E Coli ◽  
Sample Amount

Australia exports about 150,000 to 200,000 tons of manufacturing beef to the United States annually. Each lot is tested for Escherichia coli O157 using the N-60 sampling protocol, where 60 small pieces of surface meat from each lot of production are tested. A risk assessment of E. coli O157 illness from the consumption of hamburgers made from Australian manufacturing meat formed the basis to evaluate the effect of sample size and amount on the number of illnesses predicted. The sampling plans evaluated included no sampling (resulting in an estimated 55.2 illnesses per annum), the current N-60 plan (50.2 illnesses), N-90 (49.6 illnesses), N-120 (48.4 illnesses), and a more stringent N-60 sampling plan taking five 25-g samples from each of 12 cartons (47.4 illnesses per annum). While sampling may detect some highly contaminated lots, it does not guarantee that all such lots are removed from commerce. It is concluded that increasing the sample size or sample amount from the current N-60 plan would have a very small public health effect.

scholarly journals Growth of Salmonella and Other Foodborne Pathogens on Inoculated Inshell Pistachios during Simulated Delays between Hulling and Drying

2019 ◽  
Vol 82 (5) ◽  
pp. 815-825 ◽  
Author(s):  
MAHTA MOUSSAVI ◽  
VANESSA LIEBERMAN ◽  
CHRIS THEOFEL ◽  
JAVAD BAROUEI ◽  
LINDA J. HARRIS
Keyword(s):  
Escherichia Coli ◽  
Relative Humidity ◽  
Foodborne Pathogens ◽  
Lag Time ◽  
Escherichia Coli O157 ◽  
Contributing Factors ◽  
Shell Surface ◽  
E Coli ◽  
Significant Growth ◽  
Lower Maximum

ABSTRACT During harvest, pistachios are hulled, separated in water into floater and sinker streams (in large part on the basis of nut density), and then dried before storage. Higher prevalence and levels of Salmonella were previously observed in floater pistachios, but contributing factors are unclear. To examine the behavior of pathogens on hulled pistachios during simulated drying delays, floater and sinker pistachios collected from commercial processors were inoculated at 1 or 3 log CFU/g with cocktails of Salmonella and in some cases Escherichia coli O157:H7 or Listeria monocytogenes and incubated for up to 30 h at 37°C and 90% relative humidity. Populations were measured by plating onto tryptic soy agar and appropriate selective agars. In most cases, no significant growth (P > 0.05) of Salmonella was observed in the first 3 h after inoculation in hulled floaters and sinkers. Growth of Salmonella was greater on floater pistachios than on corresponding sinkers and on floater pistachios with ≥25% hull adhering to the shell surface than on corresponding floaters with <25% adhering hull. Maximum Salmonella populations (2 to 7 log CFU/g) were ∼2-log higher on floaters than on corresponding sinkers. The growth of E. coli O157:H7 and Salmonella on hulled pistachios was similar, but a longer lag time (approximately 11 h) and significantly lower maximum populations (4 versus 5 to 6 log CFU/g; P < 0.05) were predicted for L. monocytogenes. Significant growth of pathogens on hulled pistachios is possible when delays between hulling and drying are longer than 3 h, and pathogen growth is enhanced in the presence of adhering hull material.

Food Safety and Inspection Service Regulatory Testing Program for Escherichia coli O157:H7 in Raw Ground Beef

2005 ◽  
Vol 68 (3) ◽  
pp. 462-468 ◽  
Author(s):  
ALECIA LAREW NAUGLE ◽  
KRISTIN G. HOLT ◽  
PRISCILLA LEVINE ◽  
RON ECKEL
Keyword(s):  
Escherichia Coli ◽  
Food Safety ◽  
Ground Beef ◽  
Fiscal Year ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Inspection Service ◽  
Rate Ratios ◽  
Log Linear ◽  
Regulatory Actions

We analyzed raw ground beef testing data to determine whether a decrease in the rate of Escherichia coli O157:H7–positive raw ground beef samples has occurred since the inception of Food Safety and Inspection Service (U.S. Department of Agriculture) regulatory actions and microbiological testing concerning this commodity and pathogen. A main effects log-linear Poisson regression model was constructed to evaluate the association between fiscal year and the rate of E. coli O157: H7–positive raw ground beef samples while controlling for the effect of season for the subset of test results obtained from fiscal year (FY)2000 through FY2003. Rate ratios were used to compare the rate of E. coli O157:H7–positive raw ground beef samples between sequential years to identify year-to-year differences. Of the 26,521 raw ground beef samples tested from FY2000 through FY2003, 189 (0.71%) tested positive for E. coli O157:H7. Year-to-year comparisons identified a 50% reduction in the rate of positive ground beef samples from FY2002 to FY2003 when controlling for season (95% CI, 10 to 72% decrease; P = 0.02). This decrease was the only significant year-to-year change in the rate of E. coli O157:H7–positive raw ground beef samples but was consistent in samples obtained from both federally inspected establishments and retail outlets. We believe this decrease is attributed to specific regulatory actions by Food Safety and Inspection Service and subsequent actions implemented by the industry, with the goal of reducing E. coli O157:H7 adulteration of raw ground beef. Continued monitoring is necessary to confirm that the decrease in the rate of E. coli O157:H7 in raw ground beef samples we observed here represents the beginning of a sustained trend.

Inactivation of Escherichia coli O157:H7 and Salmonella and Native Microbiota on Fresh Strawberries by Antimicrobial Washing and Coating

2018 ◽  
Vol 81 (8) ◽  
pp. 1227-1235 ◽  
Author(s):  
MINGMING GUO ◽  
TONY Z. JIN ◽  
JOSHUA B. GURTLER ◽  
XUETONG FAN ◽  
MADHAV P. YADAV
Keyword(s):  
Escherichia Coli ◽  
Foodborne Pathogens ◽  
Allyl Isothiocyanate ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Native Microflora ◽  
Pathogen Populations ◽  
Antimicrobial Treatments ◽  
Antimicrobial Effectiveness

ABSTRACT Antimicrobial washing (AW), antimicrobial coating (AC), and a combination of washing followed by coating (AW+AC) were evaluated for their ability to inactivate artificially inoculated foodborne pathogens and native microbiota on strawberries stored at 4°C. Strawberries were inoculated with a six-strain composite of Escherichia coli O157:H7 and Salmonella; treated by AW, AC, or AW+AC; and stored at 4°C for 3 weeks. The washing solution contained 90 ppm of peracetic acid, and the coating solution consisted of chitosan (1%, w/v), allyl isothiocyanate (1%, v/v), and corn-bio fiber gum (5%, w/v). The effectiveness of the antimicrobial treatments against E. coli O157:H7 and Salmonella pathogens and native microflora on strawberries and their impact on fruit quality (appearance, weight loss, color, and firmness) were determined. By the end of storage, pathogen populations on strawberries were 2.5 (AW+AC), 2.9 (AC), 3.8 (AW), and 4.2 log CFU for the positive (untreated) control. AW+AC treatments also inactivated the greatest population of native microflora, followed by the AC treatment alone. AW+AC treatments showed additional antimicrobial effectiveness against these two pathogens and native microflora. Both AW+AC and AC treatments preserved the color, texture, and appearance of strawberries throughout storage. The coating treatments (AW+AC and AC alone) further reduced the loss of moisture throughout storage. The AW treatment was the least effective in reducing populations of pathogens and native microflora and in maintaining the quality of strawberries throughout storage. This study demonstrates a method to improve the microbiological safety, shelf life, and quality of strawberries.

Inactivation of Escherichia coli O157:H7 and Listeria monocytogenes on Plastic Kitchen Cutting Boards by Electrolyzed Oxidizing Water

1999 ◽  
Vol 62 (8) ◽  
pp. 857-860 ◽  
Author(s):  
KUMAR S. VENKITANARAYANAN ◽  
GABRIEL O. I. EZEIKE ◽  
YEN-CON HUNG ◽  
MICHAEL P. DOYLE
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Laminar Flow ◽  
Foodborne Pathogens ◽  
Deionized Water ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Laminar Flow Hood ◽  
Cutting Boards ◽  
Temperature Time

One milliliter of culture containing a five-strain mixture of Escherichia coli O157:H7 (∼1010 CFU) was inoculated on a 100-cm2 area marked on unscarred cutting boards. Following inoculation, the boards were air-dried under a laminar flow hood for 1 h, immersed in 2 liters of electrolyzed oxidizing water or sterile deionized water at 23°C or 35°C for 10 or 20 min; 45°C for 5 or 10 min; or 55°C for 5 min. After each temperature–time combination, the surviving population of the pathogen on cutting boards and in soaking water was determined. Soaking of inoculated cutting boards in electrolyzed oxidizing water reduced E. coli O157:H7 populations by ≥5.0 log CFU/100 cm2 on cutting boards. However, immersion of cutting boards in deionized water decreased the pathogen count only by 1.0 to 1.5 log CFU/100 cm2. Treatment of cutting boards inoculated with Listeria monocytogenes in electrolyzed oxidizing water at selected temperature–time combinations (23°C for 20 min, 35°C for 10 min, and 45°C for 10 min) substantially reduced the populations of L. monocytogenes in comparison to the counts recovered from the boards immersed in deionized water. E. coli O157:H7 and L. monocytogenes were not detected in electrolyzed oxidizing water after soaking treatment, whereas the pathogens survived in the deionized water used for soaking the cutting boards. This study revealed that immersion of kitchen cutting boards in electrolyzed oxidizing water could be used as an effective method for inactivating foodborne pathogens on smooth, plastic cutting boards.

Dispersion and Survival of Escherichia coli O157:H7 and Salmonella Typhimurium during the Production of Marinated Beef Inside Skirt Steaks and Tri-Tip Roasts

2012 ◽  
Vol 75 (2) ◽  
pp. 255-260 ◽  
Author(s):  
TIFFANY M. MURAS ◽  
KERRI B. HARRIS ◽  
LISA M. LUCIA ◽  
MARGARET D. HARDIN ◽  
JEFFREY W. SAVELL
Keyword(s):  
Escherichia Coli ◽  
Food Safety ◽  
Salmonella Typhimurium ◽  
Escherichia Coli O157 ◽  
Refrigerated Storage ◽  
E Coli ◽  
Safety Risks ◽  
Beef Products ◽  
Potential Food

To determine the depth of pathogen dispersion and the ability of pathogens to survive in enhanced beef products and spent marinade, beef inside skirt steaks and tri-tip roasts were vacuum tumbled with two commercial marinades. The marinades were inoculated with Escherichia coli O157:H7 and Salmonella Typhimurium, resulting in an approximate count of 5.2 log CFU/ml. Both inside skirt steaks and tri-tip roasts were vacuum tumbled for 1 h and sampled immediately after tumbling (day 0), or were vacuum packaged, stored (ca. 4°C), and sampled on days 7 and 14. Samples of the spent marinade were taken after tumbling (day 0) and on days 3 and 7. For both marinades, Salmonella Typhimurium and E. coli O157:H7 were dispersed throughout the inside skirt steaks during vacuum tumbling. Although Salmonella Typhimurium and E. coli O157:H7 for the skirt steaks were still detectable after 14 days of storage, the log values were lower than those on days 0 and 7. For the tri-tip roasts, the pathogen distribution varied, depending on the thickness of the roasts, and pathogens were detectable on days 0, 7, and 14. The spent marinade sampled on days 0, 3, and 7 showed that the pathogens survived at refrigerated temperatures. Because pathogens can transfer to the interior of beef inside skirt steaks and tri-tip roasts when vacuum tumbled with contaminated marinade and survived during refrigerated storage, establishments should consider the potential food safety risks associated with reuse of marinade during the production of vacuum-tumbled beef products.

Quantitative Microbial Risk Assessment for Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes in Leafy Green Vegetables Consumed at Salad Bars

2010 ◽  
Vol 73 (2) ◽  
pp. 274-285 ◽  
Author(s):  
E. FRANZ ◽  
S. O. TROMP ◽  
H. RIJGERSBERG ◽  
H. J. van der FELS-KLERX
Keyword(s):  
Public Health ◽  
Escherichia Coli ◽  
Risk Assessment ◽  
Monte Carlo ◽  
Escherichia Coli O157 ◽  
Quantitative Microbial Risk Assessment ◽  
Microbial Risk Assessment ◽  
Microbial Risk ◽  
The Public ◽  
E Coli

Fresh vegetables are increasingly recognized as a source of foodborne outbreaks in many parts of the world. The purpose of this study was to conduct a quantitative microbial risk assessment for Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes infection from consumption of leafy green vegetables in salad from salad bars in The Netherlands. Pathogen growth was modeled in Aladin (Agro Logistics Analysis and Design Instrument) using time-temperature profiles in the chilled supply chain and one particular restaurant with a salad bar. A second-order Monte Carlo risk assessment model was constructed (using @Risk) to estimate the public health effects. The temperature in the studied cold chain was well controlled below 5°C. Growth of E. coli O157:H7 and Salmonella was minimal (17 and 15%, respectively). Growth of L. monocytogenes was considerably greater (194%). Based on first-order Monte Carlo simulations, the average number of cases per year in The Netherlands associated the consumption leafy greens in salads from salad bars was 166, 187, and 0.3 for E. coli O157:H7, Salmonella, and L. monocytogenes, respectively. The ranges of the average number of annual cases as estimated by second-order Monte Carlo simulation (with prevalence and number of visitors as uncertain variables) were 42 to 551 for E. coli O157:H7, 81 to 281 for Salmonella, and 0.1 to 0.9 for L. monocytogenes. This study included an integration of modeling pathogen growth in the supply chain of fresh leafy vegetables destined for restaurant salad bars using software designed to model and design logistics and modeling the public health effects using probabilistic risk assessment software.

scholarly journals An outbreak of Escherichia coli O157 phage type 2 infection in Paisley, Scotland

2007 ◽  
Vol 12 (34) ◽  
Author(s):  
A Stirling ◽  
G McCartney ◽  
S Ahmed ◽  
J Cowden
Keyword(s):  
Public Health ◽  
Escherichia Coli ◽  
National Health ◽  
Phage Type ◽  
Health Protection ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Public Health Protection

The National Health Service’s (NHS) Greater Glasgow and Clyde Public Health Protection Unit is currently investigating an outbreak of E. coli O157 infection in Paisley.

scholarly journals Antimicrobial Resistance of Escherichia coli and Salmonella isolated from Raw Retail Broiler Chicken Carcasses in Zambia

2020 ◽  
Author(s):  
Elizabeth Muligisa Muonga ◽  
Geoffrey Mainda ◽  
Mercy Mukuma ◽  
Geoffrey Kwenda ◽  
Bernard Hang'ombe ◽  
...  
Keyword(s):  
Public Health ◽  
Escherichia Coli ◽  
Antimicrobial Resistance ◽  
Foodborne Pathogens ◽  
Broiler Chicken ◽  
Target Genes ◽  
Health Concern ◽  
Public Health Concern ◽  
E Coli ◽  
Genes Encoding

Abstract BackgroundAntimicrobial resistance (AMR) of foodborne pathogens is of public health concern, especially in developing countries such as Zambia. This study was undertaken to determine the antimicrobial resistance profiles of Escherichia coli ( E. coli ) and Salmonella isolated from raw retail broiler chicken carcasses purchased from open and supermarkets in Zambia.ResultsA total of 189 E. coli and five Salmonella isolates were isolated. Identification and confirmation of the isolates were done using Analytical Profile Index (API 20E) (Biomerieux ® ) and 16S rRNA sequencing. Antimicrobial susceptibility tests (AST) were performed using the Kirby Bauer disk diffusion technique using a panel of 10 antibiotics. Multiplex PCR was used to determine the presence of three target genes encoding for resistance: tet A, Sul 1 and bla CTX-M . WHONET 2018 software was used to analyse AST results. The E. coli isolates were mostly resistant to tetracycline (79.4%), ampicillin (51.9%), and trimethoprim/sulfamethoxazole (49.7%). Two of the five Salmonella isolates were resistant to at least one antibiotic. Forty- seven (45.2%) of the 104 isolates that were screened for the presence of the resistant genes possessed at least one of the targeted resistance genes.ConclusionThis study has demonstrated the presence of AMR E. coli and Salmonella on raw retail broiler chicken carcasses from open and supermarkets, which is of public health concern.

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