Evaluation of Modified Moore Swabs and Continuous Flow Centrifugation for Concentration of Salmonella and Escherichia coli O157:H7 from Large Volumes of Water

2011 ◽  
Vol 74 (11) ◽  
pp. 1934-1937 ◽  
Author(s):  
BLEDAR BISHA ◽  
ALMA PEREZ-MENDEZ ◽  
MICHELLE D. DANYLUK ◽  
LAWRENCE D. GOODRIDGE
Keyword(s):  
Escherichia Coli ◽  
Foodborne Pathogens ◽  
Continuous Flow ◽  
Fluorescent Proteins ◽  
Lateral Flow ◽  
Detection Methods ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Before And After ◽  
Flow Devices

Modified Moore swabs (MMS; consisting of a polyvinyl chloride cartridge filled with gauze) capture microorganisms within the packed gauze as water flows through the cartridge, while continuous flow centrifugation (CFC) uses centrifugation to sediment the microorganisms while water continuously flows in the system. This study evaluated and compared the efficacy of MMS and CFC for concentration and subsequent detection of Escherichia coli O157:H7 and Salmonella from large volumes of water (10 liters). Water samples were spiked at levels of 101,102,103, and 104 CFU/100 ml with three-strain cocktails of either E. coli O157:H7 or Salmonella serovars, which had been previously transformed with a plasmid to express resistance to ampicillin as well as green, red, or cyan fluorescent proteins. Plating was performed before and after concentration on tryptic soy agar supplemented with ampicillin in order to quantitate the concentration efficiencies of each method. The two lowest spiking levels were also enriched in low volumes of tryptic soy broth supplemented with ampicillin followed by testing via lateral flow devices. Significant (P < 0.05) concentrations of initial levels of E. coli O157:H7 in the range of 0.7 to 1.0 and 1.2 to 1.4 log were achieved within approximately 35 min of processing time via MMS and CFC, respectively. Similarly, significant (P < 0.05) concentrations were also achieved for Salmonella with 0.9 to 1.2 and 1.2 to 1.4 log concentration for MMS and CFC, respectively. There were no statistical differences (P > 0.05) between the two concentration methods in their ability to concentrate either of the two target bacteria. Significantly (P < 0.05) more spiked samples were detected by lateral flow devices following concentration and enrichment than for nonconcentrated, enriched samples. It is concluded that both MMS and CFC have potential to be used to enhance the sensitivity of downstream bacterial detection methods used to test irrigation water for the presence of foodborne pathogens.

An Aggregation-Induced Emission Material Labeling Antigen-Based Lateral Flow Immunoassay Strip for Rapid Detection of Escherichia coli O157:H7

2021 ◽  
pp. 247263032098193
Author(s):  
Cheng Liu ◽  
Shuiqin Fang ◽  
Yachen Tian ◽  
Youxue Wu ◽  
Meijiao Wu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Rapid Detection ◽  
Foodborne Pathogen ◽  
Test Strip ◽  
Lateral Flow ◽  
Detection Sensitivity ◽  
Lateral Flow Immunoassay ◽  
Escherichia Coli O157 ◽  
Aggregation Induced Emission ◽  
E Coli

Escherichia coli O157:H7 ( E. coli O157:H7) is a dangerous foodborne pathogen, mainly found in beef, milk, fruits, and their products, causing harm to human health or even death. Therefore, the detection of E. coli O157:H7 in food is particularly important. In this paper, we report a lateral flow immunoassay strip (LFIS) based on aggregation-induced emission (AIE) material labeling antigen as a fluorescent probe for the rapid detection of E. coli O157:H7. The detection sensitivity of the strip is 105 CFU/mL, which is 10 times higher than that of the colloidal gold test strip. This method has good specificity and stability and can be used to detect about 250 CFU of E. coli O157:H7 successfully in 25 g or 25 mL of beef, jelly, and milk. AIE-LFIS might be valuable in monitoring food pathogens for rapid detection.

Multiplex Quantitative PCR Assays for the Detection and Quantification of the Six Major Non-O157 Escherichia coli Serogroups in Cattle Feces†

2016 ◽  
Vol 79 (1) ◽  
pp. 66-74 ◽  
Author(s):  
P. B. SHRIDHAR ◽  
L. W. NOLL ◽  
X. SHI ◽  
B. AN ◽  
N. CERNICCHIARO ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Quantitative Pcr ◽  
Foodborne Pathogens ◽  
Virulence Genes ◽  
Target Genes ◽  
Culture Methods ◽  
Fecal Samples ◽  
E Coli ◽  
Cattle Feces ◽  
Before And After

ABSTRACT Shiga toxin–producing Escherichia coli (STEC) serogroups O26, O45, O103, O111, O121, and O145, called non-O157 STEC, are important foodborne pathogens. Cattle, a major reservoir, harbor the organisms in the hindgut and shed them in the feces. Although limited data exist on fecal shedding, concentrations of non-O157 STEC in feces have not been reported. The objectives of our study were (i) to develop and validate two multiplex quantitative PCR (mqPCR) assays, targeting O-antigen genes of O26, O103, and O111 (mqPCR-1) and O45, O121, and O145 (mqPCR-2); (ii) to utilize the two assays, together with a previously developed four-plex qPCR assay (mqPCR-3) targeting the O157 antigen and three virulence genes (stx1, stx2, and eae), to quantify seven serogroups and three virulence genes in cattle feces; and (iii) to compare the three mqPCR assays to a 10-plex conventional PCR (cPCR) targeting seven serogroups and three virulence genes and culture methods to detect seven E. coli serogroups in cattle feces. The two mqPCR assays (1 and 2) were shown to be specific to the target genes, and the detection limits were 4 and 2 log CFU/g of pure culture–spiked fecal samples, before and after enrichment, respectively. A total of 576 fecal samples collected from a feedlot were enriched in E. coli broth and were subjected to quantification (before enrichment) and detection (after enrichment). Of the 576 fecal samples subjected, before enrichment, to three mqPCR assays for quantification, 175 (30.4%) were quantifiable (≥4 log CFU/g) for at least one of the seven serogroups, with O157 being the most common serogroup. The three mqPCR assays detected higher proportions of postenriched fecal samples (P < 0.01) as positive for one or more serogroups compared with cPCR and culture methods. This is the first study to assess the applicability of qPCR assays to detect and quantify six non-O157 serogroups in cattle feces and to generate data on fecal concentration of the six serogroups.

A Comparison of the Survival in Feces and Water of Escherichia coli O157:H7 Grown under Laboratory Conditions or Obtained from Cattle Feces

2006 ◽  
Vol 69 (1) ◽  
pp. 6-11 ◽  
Author(s):  
L. SCOTT ◽  
P. McGEE ◽  
J. J. SHERIDAN ◽  
B. EARLEY ◽  
N. LEONARD
Keyword(s):  
Escherichia Coli ◽  
Foodborne Pathogen ◽  
Hemorrhagic Colitis ◽  
Contaminated Water ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Cattle Feces ◽  
Oral Inoculation ◽  
Before And After ◽  
Uremic Syndrome

Escherichia coli O157:H7 is an important foodborne pathogen that can cause hemorrhagic colitis and hemolytic uremic syndrome. Cattle feces and fecally contaminated water are important in the transmission of this organism on the farm. In this study, the survival of E. coli O157:H7 in feces and water was compared following passage through the animal digestive tract or preparation in the laboratory. Feces were collected from steers before and after oral inoculation with a marked strain of E. coli O157:H7. Fecal samples collected before cattle inoculation were subsequently inoculated with the marked strain of E. coli O157:H7 prepared in the laboratory. Subsamples were taken from both animal and laboratory-inoculated feces to inoculate 5-liter volumes of water. E. coli O157:H7 in feces survived up to 97 days, and survival was not affected by the method used to prepare the inoculating strain. E. coli O157:H7 survived up to 109 days in water, and the bacteria collected from inoculated cattle were detected up to 10 weeks longer than the laboratory-prepared culture. This study suggests that pathogen survival in low-nutrient conditions may be enhanced by passage through the gastrointestinal tract.

Use of the Systems Approach To Determine the Fate of Escherichia coli O157:H7 on Fresh Lettuce and Spinach

2009 ◽  
Vol 72 (7) ◽  
pp. 1560-1568 ◽  
Author(s):  
HELGA J. DOERING ◽  
MARK A. HARRISON ◽  
RUTH A. MORROW ◽  
WILLIAM C. HURST ◽  
WILLIAM L. KERR
Keyword(s):  
Escherichia Coli ◽  
Systems Approach ◽  
Storage Temperature ◽  
Storage Period ◽  
Categorical Variables ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Field Temperature ◽  
Before And After ◽  
Handling Practices

Lettuce and spinach inoculated with Escherichia coli O157:H7 were processed and handled in ways that might occur in commercial situations, including variations in holding times before and after product cooling, transportation conditions and temperatures, wash treatments, and product storage temperatures and times. Populations of background microflora and E. coli O157:H7 were enumerated after each step in the system. Data analysis was done to predict response variables with a combination of independent categorical variables. Field temperature, time before cooling, and wash treatment significantly affected E. coli O157:H7 populations on both products. The lowest populations of E. coli O157:H7 were encountered when precool time was minimal, lettuce was washed with chlorine, and storage temperature was 4°C. For lettuce, field and transportation temperature were not important once the storage period started, whereas after 2 days E. coli O157:H7 populations on packaged baby spinach were not affected by field temperature. On chopped iceberg lettuce and whole leaf spinach that was packaged and stored at 4°C, E. coli O157:H7 contamination could still be detected after typical handling practices, although populations decreased from initial levels in many cases by at least 1.5 log units. In abusive cases, where populations increased, the product quality quickly deteriorated. Although E. coli O157:H7 levels decreased on products handled and stored under recommended conditions, survivors persisted. This study highlights practices that may or may not affect the populations of E. coli O157:H7 on the final product.

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.

Impact of Transportation of Feedlot Cattle to the Harvest Facility on the Prevalence of Escherichia coli O157:H7, Salmonella, and Total Aerobic Microorganisms on Hides

2007 ◽  
Vol 70 (1) ◽  
pp. 17-21 ◽  
Author(s):  
A. L. REICKS ◽  
M. M. BRASHEARS ◽  
K. D. ADAMS ◽  
J. C. BROOKS ◽  
J. R. BLANTON ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Feedlot Cattle ◽  
Lower Percentage ◽  
Escherichia Coli O157 ◽  
Sample Collection ◽  
Treatment Groups ◽  
E Coli ◽  
Sampling Locations ◽  
Before And After

Prevalences of Escherichia coli O157:H7, Salmonella, and total aerobic microorganisms were determined on the hides of beef feedlot cattle before and after transport from the feedyard to the harvest facility in clean and dirty trailers. Swab samples were taken from the midline and withers of 40 animals on each of 8 days before and after shipping. After samples were collected, animals were loaded in groups of 10 on upper and lower levels of clean and dirty trailers. Animals were unloaded at the harvest facility and kept in treatment groups for sample collection after exsanguination. Salmonella was found more often on hide swabs collected from the midline than on than samples collected from the withers from animals transported in both clean and dirty trailers. Salmonella was found on significantly more hide swabs collected at harvest from both sampling locations than on those collected at the feedyard, with no differences attributed to the type of trailer. At the feedyard, clean trucks had a lower percentage of Salmonella-positive samples than did dirty trucks before animals were loaded. However, after transport, both clean and dirty trucks had a similar prevalence of Salmonella. There were no differences in Salmonella prevalence on hides collected from animals transported on the top and bottom levels of clean and dirty trucks. E. coli O157:H7 was detected on less than 2% of the samples; therefore, no practical conclusions about prevalence could be drawn. Hides sampled at harvest had higher concentrations of aerobic microorganisms than did hides sampled at the feedyard, and concentrations were higher on the midline than on the withers. Although the prevalences of Salmonella and total aerobic microorganisms increased on hides after transport from the feedyard to the plant, this increase was not related to the cleanliness of the trailers or the location of the cattle in the trailers.

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.

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.

Comparative Evaluation of a Phage Protein Ligand Assay with Real-Time PCR and a Reference Method for the Detection of Escherichia coli O157:H7 in Raw Ground Beef and Trimmings

2011 ◽  
Vol 74 (1) ◽  
pp. 6-12 ◽  
Author(s):  
F. SAVOYE ◽  
P. FENG ◽  
C. ROZAND ◽  
M. BOUVIER ◽  
A. GLEIZAL ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Real Time ◽  
Ground Beef ◽  
Reference Method ◽  
Enterohemorrhagic Escherichia Coli ◽  
Detection Methods ◽  
Escherichia Coli O157 ◽  
Rt Pcr ◽  
Raw Meat ◽  
E Coli

Enterohemorrhagic Escherichia coli O157:H7 is an important pathogen associated with infections caused by consumption of undercooked raw meat. Sensitive and rapid detection methods for E. coli O157:H7 are essential for the meat industry to ensure a safe meat supply. This study was conducted to compare the sensitivity of the VIDAS ultraperformance E. coli test (ECPT UP) with a noncommercial real-time (RT) PCR method and the U.S. Department of Agriculture, Food Safety and Inspection Service (USDA-FSIS) reference method for detecting E. coli O157:H7 in raw ground beef. Optimal enrichment times and the efficacy of testing different types of raw meat, either as individual samples (25 g) or as composites (375 g), were examined. For 25-g samples of each type of raw ground beef tested, 6 h of enrichment was sufficient for both the VIDAS ECPT UP and RT-PCR methods, but for 375-g samples, 24 h of enrichment was required. Both the VIDAS ECPT UP and RT-PCR methods produced results similar to those obtained with the USDA-FSIS reference method after 18 to 24 h of enrichment. The primer specificity of the RT-PCR assay and the highly specific phage ligand used in the VIDAS ECPT UP for target recognition enabled the detection of low levels of E. coli O157:H7 in 25 g of various types of raw ground beef. The tests also allowed the detection of E. coli O157:H7 in composite raw ground beef and trimmings in samples of up to 375 g.

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.

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