Inoculation of Beef with Low Concentrations of Escherichia coli O157:H7 and Examination of Factors That Interfere with Its Detection by Culture Isolation and Rapid Methods†

2010 ◽  
Vol 73 (12) ◽  
pp. 2180-2188 ◽  
Author(s):  
JOSEPH M. BOSILEVAC ◽  
NORASAK KALCHAYANAND ◽  
JOHN W. SCHMIDT ◽  
STEVEN D. SHACKELFORD ◽  
TOMMY L. WHEELER ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Immunomagnetic Separation ◽  
Fat Content ◽  
Plate Count ◽  
Escherichia Coli O157 ◽  
Rapid Methods ◽  
E Coli ◽  
Low Concentrations ◽  
Commercial Method ◽  
Aerobic Plate Count

Currently used industry testing programs require the ability to detect Escherichia coli O157:H7 in samples of beef trim or ground beef at levels as low as 1 CFU/375 g. We present a reliable protocol for generating a control inoculum for verification testing at this low concentration and evaluate its use. Results show that half of all samples received no cells when 1 CFU was the target concentration and that targets greater than 3 CFU were much more reliable. Detection by culture isolation and two commercial assays, Qualicon BAX-MP and BioControl GDS, detected 94% ± 11%, 92% ± 10%, and 92% ± 7% of samples inoculated with 5.4 CFU (range 1 to 9 CFU), respectively. We also examined the effect of background aerobic plate count (APC) bacteria and fat content effects on the detection of E. coli O157:H7. At APC concentrations below 6 log CFU/g, the rapid methods detected all beef trim samples inoculated with 26 CFU of E. coli O157:H7 per 65 g. At an APC of 6.7 log CFU/g, culture, BAX-MP, and GDS detected 100, 75, and 13%, respectively, of inoculated samples. Neither commercial method detected E. coli O157:H7 in the samples when APC was 7.7 log CFU/g, whereas culture was able to detect 63% of E. coli O157:H7 in the samples when APC was at this concentration. Increased fat content correlated with decreasing recovery of immunomagnetic separation beads, but this was not observed to interfere with detection of E. coli O157:H7.

Fate of Enterohemorrhagic Escherichia coli O157: H7 in Commercial Mayonnaise

1994 ◽  
Vol 57 (9) ◽  
pp. 780-783 ◽  
Author(s):  
TONG ZHAO ◽  
MICHAEL P. DOYLE
Keyword(s):  
Escherichia Coli ◽  
Enterohemorrhagic Escherichia Coli ◽  
Health Concern ◽  
Plate Count ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Initial Ph ◽  
Contaminated Food ◽  
Aerobic Plate Count ◽  
Mold And Yeast

The fate of enterohemorrhagic Escherichia coli O157:H7 was determined in three different lots of commercial mayonnaise, including four different samples from a lot implicated in an outbreak of E. coli O157:H7 infection. The initial pH of the products ranged from 3.6 to 3.9. Products were inoculated with 6.5 × 103 E. coli O157:H7/g and incubated at 5 or 20°C. Escherichia coli O157:H7 did not grow at either temperature but survived for 34 to 55 days at 5°C and for 8 to 21 days at 20°C, depending on the lot. Survival was greatest in real mayonnaise purchased at retail among six mayonnaise samples which included a reduced calorie mayonnaise. Escherichia coli O157:H7 populations decreased between 2- and 100-fold by 3 weeks at 5°C, and between 10- and 1,000-fold by 7 days at 20°C. There was little or no change in pH (<0.1 unit), aerobic plate count, mold and yeast count or Lactobacillus count (< 1 log10 CFU/g) for the duration of the study. Commercial mayonnaise manufactured under good manufacturing practices is not a public health concern. Abusive handling of mayonnaise resulting in cross-contamination with E. coli O157:H7-contaminated food or contamination by an infected foodhandler is the principal basis for concern.

Surface and Internalized Escherichia coli O157:H7 on Field-Grown Spinach and Lettuce Treated with Spray-Contaminated Irrigation Water

2010 ◽  
Vol 73 (6) ◽  
pp. 1023-1029 ◽  
Author(s):  
MARILYN C. ERICKSON ◽  
CATHY C. WEBB ◽  
JUAN CARLOS DIAZ-PEREZ ◽  
SHARAD C. PHATAK ◽  
JOHN J. SILVOY ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Irrigation Water ◽  
Enrichment Culture ◽  
Field Studies ◽  
Plate Count ◽  
Escherichia Coli O157 ◽  
Adaxial Side ◽  
Abaxial Side ◽  
E Coli ◽  
Contaminated Irrigation Water

Numerous field studies have revealed that irrigation water can contaminate the surface of plants; however, the occurrence of pathogen internalization is unclear. This study was conducted to determine the sites of Escherichia coli O157:H7 contamination and its survival when the bacteria were applied through spray irrigation water to either field-grown spinach or lettuce. To differentiate internalized and surface populations, leaves were treated with a surface disinfectant wash before the tissue was ground for analysis of E. coli O157:H7 by direct plate count or enrichment culture. Irrigation water containing E. coli O157:H7 at 102, 104, or 106 CFU/ml was applied to spinach 48 and 69 days after transplantation of seedlings into fields. E. coli O157:H7 was initially detected after application on the surface of plants dosed at 104 CFU/ml (4 of 20 samples) and both on the surface (17 of 20 samples) and internally (5 of 20 samples) of plants dosed at 106 CFU/ml. Seven days postspraying, all spinach leaves tested negative for surface or internal contamination. In a subsequent study, irrigation water containing E. coli O157:H7 at 108 CFU/ml was sprayed onto either the abaxial (lower) or adaxial (upper) side of leaves of field-grown lettuce under sunny or shaded conditions. E. coli O157:H7 was detectable on the leaf surface 27 days postspraying, but survival was higher on leaves sprayed on the abaxial side than on leaves sprayed on the adaxial side. Internalization of E. coli O157:H7 into lettuce leaves also occurred with greater persistence in leaves sprayed on the abaxial side (up to 14 days) than in leaves sprayed on the adaxial side (2 days).

Antibacterial Effect of Monocaprylin on Escherichia coli O157:H7 in Apple Juice

2005 ◽  
Vol 68 (9) ◽  
pp. 1895-1899 ◽  
Author(s):  
MANOJ KUMAR MOHAN NAIR ◽  
HANEM ABOUELEZZ ◽  
THOMAS HOAGLAND ◽  
KUMAR VENKITANARAYANAN
Keyword(s):  
Escherichia Coli ◽  
Apple Juice ◽  
Antibacterial Effect ◽  
Storage Temperature ◽  
Escherichia Coli O157 ◽  
Organoleptic Properties ◽  
E Coli ◽  
Low Concentrations ◽  
Storage Temperatures ◽  
Control Samples

The antibacterial effect of low concentrations of monocaprylin on Escherichia coli O157:H7 in apple juice was investigated. Apple juice alone (control) or containing 2.5 mM (0.055%) or 5 mM monocaprylin was inoculated with a five-strain mixture of E. coli O157:H7 at ~6.0 log CFU/ml. The juice samples were stored at 23 or 4°C for 14 or 21 days, respectively, and the population of E. coli O157:H7 was determined on tryptic soy agar plates supplemented with 0.6% yeast extract. At both storage temperatures, the population of E. coli O157:H7 in monocaprylin-supplemented juice samples was significantly lower (P < 0.05) than that in the control samples. The concentration of monocaprylin and the storage temperature had a significant effect on the inactivation of E. coli O157:H7 in apple juice. Monocaprylin at 5 mM was significantly more effective than 2.5 mM monocaprylin for killing E. coli O157:H7 in apple juice. Inactivation of E. coli O157:H7 by monocaprylin was more pronounced in juice stored at 23°C than in the refrigerated samples. Results of this study indicated that monocaprylin is effective for killing E. coli O157:H7 in apple juice, but detailed sensory studies are needed to determine the organoleptic properties of apple juice containing monocaprylin.

Bioluminescence: A Rapid Indicator of Escherichia Coli O157:H7 in Selected Yogurt and Cheese Varieties

1997 ◽  
Vol 60 (8) ◽  
pp. 891-897 ◽  
Author(s):  
L. M. HUDSON ◽  
J. CHEN ◽  
A. R. HILL ◽  
M. W. GRIFFITHS
Keyword(s):  
Escherichia Coli ◽  
Enterohemorrhagic Escherichia Coli ◽  
Cottage Cheese ◽  
Plate Count ◽  
Escherichia Coli O157 ◽  
Potential Hazard ◽  
Growth And Survival ◽  
E Coli ◽  
Standard Plate Count ◽  
Standard Plate

Outbreaks of enterohemorrhagic Escherichia coli O157:H7 have been commonly associated with products derived from ground beef, but recently the organism has been implicated as the causative agent in outbreaks involving yogurt and cheese. This finding has raised concern about the potential for its growth and survival in fermented dairy products. A bioluminescent strain of E. coli O157:H7 was used to determine postprocessing survival in yogurt with live cultures at pH 4.17, 4.39, and 4.47 stored at 4 and 10°C. In addition, survival of E. coli O157:H7 was monitored during the manufacture of Cottage, Colby, Romano, and Feta cheeses. Results indicated survival for 8 and 5 days at 4 and 10°C respectively in yogurt at pH 4.17, 17 and 15 days at 4 and 10°C respectively in yogurt at pH 4.39, and 17days at both 4 and 10°C in yogurt at pH 4.47. E. coli O157:H7 did not survive cooking procedures at 56°C in Cottage cheese. However, the pathogen survived for 27, 30, and 27 days in Colby, Romano, and Feta cheeses respectively. A high correlation of r2 > 0.89 was obtained between counts of bioluminescenct colonies and standard plate count for all yogurt and cheese varieties, indicating that bioluminescence was a sensitive and rapid indicator of cellular viability for E. coli O157:H7. Survival of the pathogen, as indicated by this method, is possible in highly acidic environments even at refrigeration temperatures. This poses a potential hazard should postprocessing contamination occur.

Comparison of Rapid Enzyme-Linked Immunosorbent Assay and Immunomagnetic Separation Methods for Detection of Escherichia coli O157 in Fecal, Hide, Carcass, and Ground Beef Samples

2007 ◽  
Vol 70 (10) ◽  
pp. 2230-2234 ◽  
Author(s):  
T. W. THOMPSON ◽  
T. P. STEPHENS ◽  
G. H. LONERAGAN ◽  
M. F. MILLER ◽  
M. M. BRASHEARS
Keyword(s):  
Escherichia Coli ◽  
Ground Beef ◽  
Immunomagnetic Separation ◽  
Separation Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Escherichia Coli O157 ◽  
Perfect Agreement ◽  
Fecal Samples ◽  
E Coli ◽  
Beef Products

Rapid enzyme-linked immunosorbent assays (ELISAs) are approved for detection of Escherichia coli O157 in beef products. However, these kits have also been used in the industry to detect this pathogen on hides or in feces of cattle, although this use has not been validated. The objective of this study was to compare commercially available ELISAs (E. coli Now, Reveal, and VIP) with immunomagnetic separation along with selective media to detect E. coli O157 on hides, in feces, and in medium- and low-level-inoculated ground beef and carcasses (simulated by using briskets) samples. Naturally infected hide and fecal samples were subjected to both the immunomagnetic separation method and ELISAs for the detection of E. coli O157. Additionally, E. coli O157 inoculated and noninoculated ground beef and beef briskets were used to simulate meat and carcass samples. When comparing the detection results from the ELISAs (E. coli Now, Reveal, and VIP) to the immunomagnetic separation method, poor agreement was observed for fecal samples (kappa = 0.10, 0.02, and 0.03 for E. coli Now, Reveal, and VIP, respectively), and fair-to-moderate agreement was observed for hide samples (kappa = 0.30, 0.51, and 0.29 for E. coli Now, Reveal, and VIP, respectively). However, there was near-perfect agreement between the immunomagnetic separation method and ELISAs for ground beef (kappa = 1, 1, and 0.80 for E. coli Now, Reveal, and VIP, respectively) and brisket (kappa = 1, 1, and 1 for E. coli Now, Reveal, and VIP, respectively) samples. Assuming immunomagnetic separation is the best available method, these data suggest that the ELISAs are not useful in detecting E. coli O157 from hide or fecal samples. However, when ELISAs are used on ground beef and beef brisket samples they can be used with a high degree of confidence.

Contamination Revealed by Indicator Microorganism Levels during Veal Processing

2016 ◽  
Vol 79 (8) ◽  
pp. 1341-1347 ◽  
Author(s):  
JOSEPH M. BOSILEVAC ◽  
RONG WANG ◽  
BRANDON E. LUEDTKE ◽  
TOMMY L. WHEELER ◽  
MOHAMMAD KOOHMARAIE
Keyword(s):  
Escherichia Coli ◽  
Plate Count ◽  
Department Of Agriculture ◽  
E Coli ◽  
Veal Calves ◽  
Indicator Microorganism ◽  
Analysis Of Results ◽  
Aerobic Plate Count ◽  
Inspection Service ◽  
The U.S

ABSTRACT During site visits of veal processors, the U.S. Department of Agriculture, Food Safety Inspection Service (FSIS) has reported processing deficiencies that likely contribute to increased levels of veal contamination. Here, we report the results of measuring aerobic plate count bacteria (APC), Enterobacteriaceae, coliforms (CF), and Escherichia coli during eight sample collections at five veal processors to assess contamination during the harvest of bob veal and formula-fed veal before (n = 5 plants) and after (n = 3 plants) changes to interventions and processing practices. Hides of veal calves at each plant had mean log CFU/100 cm2 APC, Enterobacteriaceae, CF, and E. coli of 6.02 to 8.07, 2.95 to 5.24, 3.28 to 5.83, and 3.08 to 5.59, respectively. Preintervention carcasses had mean log CFU/100 cm2 APC, Enterobacteriaceae, CF, and E. coli of 3.08 to 5.22, 1.16 to 3.47, 0.21 to 3.06, and −0.07 to 3.10, respectively, before and 2.72 to 4.50, 0.99 to 2.76, 0.69 to 2.26, and 0.33 to 2.12, respectively, after changes were made to improve sanitary dressing procedures. Final veal carcasses had mean log CFU/100 cm2 APC, Enterobacteriaceae, CF, and E. coli of 0.36 to 2.84, −0.21 to 1.59, −0.23 to 1.59, and −0.38 to 1.45 before and 0.44 to 2.64, −0.16 to 1.33, −0.42 to 1.20, and −0.48 to 1.09 after changes were made to improve carcass-directed interventions. Whereas the improved dressing procedures resulted in improved carcass cleanliness, the changes to carcass-directed interventions were less successful, and veal processors are urged to use techniques that ensure uniform and consistent delivery of antimicrobials to carcasses. Analysis of results comparing bob veal to formula-fed veal found bob veal hides, preintervention carcasses, and final carcasses to have increased (P < 0.05) APC, Enterobacteriaceae, CF, and E. coli (with the exception of hide Enterobacteriaceae; P > 0.05) relative to formula fed veal. When both veal categories were harvested at the same plant on the same day, similar results were observed. Since identification by FSIS, the control of contamination during veal processing has started to improve, but challenges still persist.

Standardized Microbiological Sampling and Testing Procedures for the Beef Industry†

1996 ◽  
Vol 59 (7) ◽  
pp. 778-780 ◽  
Author(s):  
KELLY J. KARR ◽  
ELIZABETH A. E. BOYLE ◽  
CURTIS L. KASTNER ◽  
JAMES L. MARSDEN ◽  
RANDALL K. PHEBUS ◽  
...  
Keyword(s):  
Total Coliform ◽  
Plate Count ◽  
Escherichia Coli O157 ◽  
Beef Industry ◽  
Salmonella Spp ◽  
Testing Procedures ◽  
E Coli ◽  
Aerobic Plate Count ◽  
Reported Data ◽  
Microbiological Analyses

Standardized microbiological sampling and testing procedures were developed that can be used throughout the beef slaughter and processing industry to facilitate the collection and any desired compilation of comparative data. Twenty samples each from carcasses (brisket, flank, and rump areas combined); subprimal cuts (clods); lean trim; and cutting and/or conveyor surfaces were collected in three slaughter and processing operations, with the first operation being a preliminary trial and resulting in no reported data. Microbiological analyses for Clostridium perfringens, Escherichia coli O157:H7, Listeria monocytogenes, Salmonella spp., Staphylococcus aureus, Campylobacter jejuni/coli, total coliforms, E. coli Biotype I, and aerobic mesophilic bacteria (aerobic plate count, APC) were performed on all samples by an outside laboratory. The procedures developed were effective in allowing samples to be collected, shipped, and analyzed in the same manner for all operations. From a logistical standpoint, approximately 20 samples each of carcasses, clods, lean trim, and surfaces could be taken within 4 to 6 h by five people. Forty samples each of carcass, clod, lean trim, and conveyor surfaces from two plants tested negative for E. coli O157:H7, Salmonella spp., and Listeria spp., with the exception of L. monocytogenes being isolated from one carcass and one clod sample. APCs and total coliform counts were between 103 to 105 and 102 to 103 CFU/cm2 or CFU/g, respectively, for the 40 samples each of carcasses, clods, and lean trim. APCs for surface swab counts ranged from ≤ 10 to 103 CFU/cm2.

scholarly journals Development and Characterization of a Fluorescent-Bacteriophage Assay for Detection of Escherichia coli O157:H7

1999 ◽  
Vol 65 (4) ◽  
pp. 1397-1404 ◽  
Author(s):  
Lawrence Goodridge ◽  
Jinru Chen ◽  
Mansel Griffiths
Keyword(s):  
Escherichia Coli ◽  
Flow Cytometry ◽  
Immunomagnetic Separation ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Lower Detection ◽  
Unreliable Method ◽  
Direct Epifluorescent Filter Technique ◽  
Bacterial Concentrations

ABSTRACT In this paper we describe evaluation and characterization of a novel assay that combines immunomagnetic separation and a fluorescently stained bacteriophage for detection of Escherichia coliO157:H7 in broth. When it was combined with flow cytometry, the fluorescent-bacteriophage assay (FBA) was capable of detecting 104 cells/ml. A modified direct epifluorescent-filter technique (DEFT) was employed in an attempt to estimate bacterial concentrations. Using regression analysis, we calculated that the lower detection limit was between 102 and 103cells/ml; however, the modified DEFT was found to be an unreliable method for determining bacterial concentrations. The results of this study show that the FBA, when combined with flow cytometry, is a sensitive technique for presumptive detection of E. coliO157:H7 in broth cultures.

Decontamination of Beef Subprimal Cuts Intended for Blade Tenderization or Moisture Enhancement

2007 ◽  
Vol 70 (5) ◽  
pp. 1174-1180 ◽  
Author(s):  
C. E. HELLER ◽  
J. A. SCANGA ◽  
J. N. SOFOS ◽  
K. E. BELK ◽  
W. WARREN-SERNA ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Lactic Acid ◽  
Hot Water ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Low Concentrations ◽  
Blade Tenderization ◽  
Antimicrobial Interventions

The prevalence of Escherichia coli O157:H7 on beef subprimal cuts intended for mechanical tenderization was evaluated. This evaluation was followed by the assessment of five antimicrobial interventions at minimizing the risk of transferring E. coli O157:H7 to the interior of inoculated subprimal cuts during blade tenderization (BT) or moisture enhancement (ME). Prevalence of E. coli O157:H7 on 1,014 uninoculated beef subprimals collected from six packing facilities was 0.2%. Outside round pieces inoculated with E. coli O157:H7 at 104 CFU/100 cm2 were treated with (i) no intervention, (ii) surface trimming, (iii) hot water (82°C), (iv) warm 2.5% lactic acid (55°C), (v) warm 5.0% lactic acid (55°C), or (vi) 2% activated lactoferrin followed by warm 5.0% lactic acid (55°C) and then submitted to BT or ME. Prevalence (n = 196) of internalized (BT and ME) E. coli O157:H7 was 99%. Enumeration of E. coli O157:H7 (n = 192) revealed mean surface reductions of 0.93 to 1.10 log CFU/100 cm2 for all antimicrobial interventions. E. coli O157:H7 was detected on 3 of the 76 internal BT samples and 73 of the 76 internal ME samples. Internal ME samples with no intervention had significantly higher mean E. coli O157:H7 populations than did those internal samples treated with an intervention, but there were no significant differences in E. coli O157:H7 populations among internal BT samples. Results of this study demonstrate that the incidence of E. coli O157:H7 on the surface of beef subprimal cuts is low and that interventions applied before mechanical tenderization can effectively reduce the transfer of low concentrations of E. coli O157:H7 to the interior of beef subprimal cuts.

Survival of Escherichia coli O157:H7 in Ground-Beef Patties during Storage at 2, −2, 15 and then −2°C, and −20°C†

1999 ◽  
Vol 62 (11) ◽  
pp. 1243-1247 ◽  
Author(s):  
SUSAN E. ANSAY ◽  
KIM A. DARLING ◽  
CHARLES W. KASPAR
Keyword(s):  
Escherichia Coli ◽  
Frozen Storage ◽  
Ground Beef ◽  
Plate Count ◽  
Escherichia Coli O157 ◽  
Control Strain ◽  
Single Strain ◽  
E Coli ◽  
Beef Patties ◽  
Significant Difference

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.

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