Direct Enumeration of Escherichia coli O157:H7 from Petrifilm™ EC Count Plates Using the Petrifilm™ Test Kit — HEC Without Sample Pre-Enrichment

1994 ◽  
Vol 57 (10) ◽  
pp. 859-864 ◽  
Author(s):  
MELISSA L. CALICCHIA ◽  
JANICE D. REGER ◽  
CONNIE I. N. WANG ◽  
DARYL W. OSATO
Keyword(s):  
Escherichia Coli ◽  
Package Insert ◽  
Raw Milk ◽  
Total Coliform ◽  
Food Samples ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Surface Samples ◽  
Test Kit ◽  
Cutting Boards

Enumeration of Escherichia coli O157:H7 from Petrifilm™ E. coli (PEC) Count plates was studied incorporating the Petrifilm™ -HEC kit without sample pre-enrichment. Samples containing various E. coli O157:H7 inoculum levels were plated according to PEC package insert instructions. Plates were incubated at 32°C for dairy products and at 35°C for other samples. Total coliform, E. coli and E. coli O157:H7 levels were quantified in both control and inoculated samples. The average recovery from various food matrices was 74.8%. Food samples included retail soft cheese, clams, ground beef, chicken, salad mix, apple juice, cantaloupe and raw milk. The overall recovery was also determined on surface samples including cutting boards and preparation tables, at 116.3%. Results suggest that enumeration of E. coli O157:H7 from PEC Count plates is possible for products containing background coliforms as high as 40,000 colony forming units (CFU)/g, and with minimal background coliforms at <10 CFU/g. This method is a fast and efficient means of quantifying levels of presumptive positive E. coli O157:H7 from PEC Count plates.

Influence of Lactic Acid Bacteria on Survival of Escherichia coli O157:H7 in Inoculated Minas Cheese during Storage at 8.5°C

2001 ◽  
Vol 64 (8) ◽  
pp. 1151-1155 ◽  
Author(s):  
SUSANA M. I. SAAD ◽  
CÉZAR VANZIN ◽  
MARICÊ N. OLIVEIRA ◽  
BERNADETTE D. G. M. FRANCO
Keyword(s):  
Escherichia Coli ◽  
Hazard Analysis ◽  
Control Point ◽  
Raw Milk ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Critical Control Point ◽  
Plastic Bags ◽  
Test Kit ◽  
Sterile Plastic

Minas cheese is a typical Brazilian fresh cheese, manufactured by addition of rennin and CaCl2 to milk, followed by draining the curd. The intrinsic characteristics of this product make it favorable for growth of pathogens, including Escherichia coli O157:H7. The influence of the addition of a commercial mesophilic type O lactic culture to this product on the growth of this pathogen during storage at 8.5°C was evaluated. Eight different formulations of Minas cheese were manufactured using raw or pasteurized milk and with or without salt and lactic culture. Individual portions of each formulation were transferred to sterile plastic bags and inoculated with E. coli O157:H7 to yield ca. 103 or 106 CFU/g. After blending by hand massaging the bags, samples were stored at 8.5°C for up to 14 days. E. coli O157:H7 was counted after 1, 2, 7, and 14 days of storage using 3M Petrifilm Test Kit-HEC. Counts in samples without added lactic culture showed a 2-log increase in the first 24 h and remained constant during the following 14 days. Counts in samples with added lactic culture showed a 0.5-log increase in the first 24 h, followed by a decrease. These variations were statistically significant (P < 0.05). No significant variations (P > 0.05) were obtained for cheese samples manufactured with pasteurized or raw milk, with or without salt. Results indicate that the addition of type O lactic culture may be an additional safeguard to well-established good manufacturing practices and hazard analysis and critical control point programs in the control of growth of E. coli O157:H7 in Minas cheese.

Evaluation of the VIDAS Methodology for Detection of Escherichia coli O157 in Food Samples

1998 ◽  
Vol 61 (7) ◽  
pp. 917-920 ◽  
Author(s):  
C. VERNOZY-ROZAND ◽  
C. MAZUY ◽  
S. RAY-GUENIOT ◽  
S. BOUTRAND-LOEÏ ◽  
A. MEYRAND ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Raw Milk ◽  
Immunomagnetic Separation ◽  
Food Samples ◽  
Positive Sample ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Contaminated Food ◽  
Capture And Release ◽  
Retail Food

An automated enzyme-linked fluorescence immunoassay (ELFA), the VIDAS E. coli O157 method, was compared with immunomagnetic separation (IMS) followed by culture on cefixime tellurite sorbitol MacConkey agar (CT-SMAC) for detecting Escherichia coli O157 in artificially and naturally contaminated food samples including raw milk cheeses, poultry, raw sausages, and ground beef retail samples. Confirmation of the samples positive according to the ELFA was performed by use of an automated immunoconcentration system, VIDAS ICE, which allows selective capture and release of target organisms. A total of 496 retail food samples were examined. Seventeen food samples gave positive values with the ELFA method, and among them 9 food samples were confirmed by the ICE method. Eight were shown to contain sorbitol-positive, O157-positive, H7-negative, motile, non-verotoxin-producing E. coli. The ninth positive sample contained an O157-positive, H7-negative, sorbitol-negative, non-verotoxin-producing E. coli. The IMS technique only allowed confirmation of this sorbitol-negative, non-verotoxin-producing E. coli O157.

Simultaneous Enrichment of Shiga Toxin–Producing Escherichia coli O157 and O26 and Salmonella in Food Samples Using Universal Preenrichment Broth

2009 ◽  
Vol 72 (10) ◽  
pp. 2065-2070 ◽  
Author(s):  
MASASHI KANKI ◽  
KAZUKO SETO ◽  
JUNKO SAKATA ◽  
TETSUYA HARADA ◽  
YUKO KUMEDA
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Food Samples ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Significant Difference ◽  
Peptone Water ◽  
Radish Sprouts ◽  
Pork Samples ◽  
Better Than

Universal preenrichment broth (UPB) was compared with modified Escherichia coli broth with novobiocin (mEC+n) for enrichment of Shiga toxin–producing E. coli O157 and O26, and with buffered peptone water (BPW) for preenrichment of Salmonella enterica. Ten strains each of the three pathogens were inoculated into beef and radish sprouts following thermal, freezing, or no treatment. With regard to O157 and O26, UPB incubated at 42°C recovered significantly more cells from inoculated beef than UPB at 35°C and from radish sprout samples than UPB at 35°C and mEC+n. With regard to Salmonella, UPB incubated at 42°C was as effective as UPB at 35°C and BPW at recovering cells from beef and radish sprout samples. No significant difference was noted between the effectiveness of UPB at 42°C and UPB at 35°C or BPW in the recovery of Salmonella from 205 naturally contaminated poultry samples. By using UPB at 42°C, one O157:H7 strain was isolated from the mixed offal of 53 beef samples, 6 cattle offal samples, and 50 pork samples all contaminated naturally, with no pathogen inoculation. The present study found that UPB incubated at 42°C was as effective as, or better than, mEC+n for enrichment of O157 and O26 and comparable to BPW for preenrichment of Salmonella. These findings suggest that a great deal of labor, time, samples, and space may be saved if O157, O26, and Salmonella are enriched simultaneously with UPB at 42°C.

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.

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.

Comparison of the SimPlate Coliform and Escherichia coli Test with Petrifilm, Three-Tube MPN, and VRBA + MUG Methods for Enumerating Coliforms and E. coli in Food

1998 ◽  
Vol 61 (4) ◽  
pp. 444-449 ◽  
Author(s):  
D. E. TOWNSEND ◽  
R. L. IRVING ◽  
A. NAQUI
Keyword(s):  
Escherichia Coli ◽  
Correlation Coefficients ◽  
Raw Milk ◽  
Food Samples ◽  
Coliform Bacteria ◽  
Most Probable Number ◽  
Additional Food ◽  
Suitable Alternative ◽  
Probable Number ◽  
E Coli

SimPlate for coliforms and Escherichia coli (CEc) is a new method for the detection and quantification of coliforms and E. coli in food. Internal validation of the method was carried out at IDEXX Laboratories (Westbrook, ME) with 180 food samples representing a variety of different food matrices and compared against three-tube MPN (most probable number), VRBA (violet red bile agar) + MUG, and Petrifilm (E. coli count) methods. SimPlate CEc was highly correlated with each of these methods for the quantification of coliform bacteria (r ≥ 0.90). An insignificant number of food samples were found to contain E. coli; therefore, no meaningful correlation data could be generated. Four hundred forty-four additional food samples were tested at five collaborating laboratories for the presence of coliforms and E. coli using SimPlate CEc and either VRBA + MUG or Petrifilm (E. coli count). Regression analysis of data from SimPlate for CEc versus Petrifilm E. coli count plates generated correlation coefficients (r) of at least 0.89 for total coliforms and at least 0.90 for generic E. coli. Correlation coefficients between SimPlate for CEc and VRBA + MUG data were at least 0.90 for coliforms and at least 0.86 for E. coli. SimPlate for CEc demonstrated better recovery of E. coli than Petrifilm when high populations of bacteria were present. E. coli was not detected in 20 of 50 (40%) raw milk samples tested by the Petrifilm method due to the presence of interfering coliform and noncoliform bacteria. It is concluded that SimPlate for CEc is a suitable alternative for determining numbers of coliform bacteria and E. coli in food.

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.

Testing of Bob Calf Fecal Swabs for the Presence of Escherichia coli O157:H7

1994 ◽  
Vol 57 (1) ◽  
pp. 70-72 ◽  
Author(s):  
DAVID R. MARTIN ◽  
PAUL M. UHLER ◽  
ANITA J. G. OKREND ◽  
JOSEPH Y. CHIU
Keyword(s):  
Escherichia Coli ◽  
The Other ◽  
Escherichia Coli O157 ◽  
Enrichment Method ◽  
Direct Smear ◽  
E Coli ◽  
Test Kit ◽  
Rectal Swabs

Rectal swabs were collected from 304 Bob calves (calves under 10 d old) brought to slaughter in the States of Washington (77 swabs), California (127 swabs), and Wisconsin (100 swabs). The swab samples were tested for the presence of Escherichia coli O157:H7 by the use of a direct smear method, enrichment method, and the use of the Petrifilm™ Test Kit-HEC-for hemorrhagic E. coli O157:H7 (3M Company, St. Paul, MN). The organism was not isolated from any of the samples by any method, though the 3M test kit did give 21 positive signals. Of these positive signals, three were shown to be caused by sorbitol-positive, O157-positive, H7-negative E. coli. The cause of the other 18 signals was not determined.

Prevalence of Escherichia coli O157:H7 and Indicator Organisms on the Surface of Intact Subprimal Beef Cuts Prior to Further Processing

2006 ◽  
Vol 69 (7) ◽  
pp. 1514-1517 ◽  
Author(s):  
JAMES E. KENNEDY ◽  
SALLY K. WILLIAMS ◽  
TED BROWN ◽  
PHIL MINERICH
Keyword(s):  
Escherichia Coli ◽  
Total Coliform ◽  
Escherichia Coli O157 ◽  
Indicator Organisms ◽  
Seasonal Differences ◽  
Total Coliforms ◽  
E Coli ◽  
Plate Counts ◽  
Collection Period

The objective of this study was to determine the prevalence of Escherichia coli O157:H7, other E. coli strains, total coliforms, and aerobic organisms on the surface of subprimal beef cuts prior to enhancement. Subprimal cuts were sampled during winter (January and February 2004) and summer (August through October 2004). During each collection period, six representative subprimal cuts (chuck tenders, 0.64-cm trimmed strips, bottom round flat, rough-trimmed brisket, cap-on top rounds, and cap-off insides) were sampled. A total of 600 samples in winter (100 samples per cut) and 599 samples in summer (100 chuck tenders, 100 0.64-cm trimmed strips, 100 bottom round flats, 100 cap-off insides, 97 rough-trimmed briskets, and 102 cap-on top rounds) were collected from five plants in the Midwest, southern Midwest, northern Midwest, and Southeast and swabbed using the sponge swab method. All sponges were analyzed for E. coli O157:H7. In addition, 400 subprimal cuts from four plants were analyzed for aerobic plate counts, total coliforms, and other E. coli strains during each collection period. E. coli O157:H7 was not detected on any of the 1,199 subprimal samples; thus, incidence of E. coli O157:H7 was <0.083%. Seasonal differences between aerobic plate counts and total coliform counts for each of the same cuts were 1.0 log CFU per cut or less. E. coli strains were not detected in 82, 52, 69, and 82% of the chuck tenders, 0.64-cm trimmed strips, bottom round flats, and cap-off insides, respectively.

Enumeration of Escherichia coli O157 in Outbreak-Associated Gouda Cheese Made with Raw Milk

2015 ◽  
Vol 78 (9) ◽  
pp. 1733-1737 ◽  
Author(s):  
ALEXANDER GILL ◽  
DENISE OUDIT
Keyword(s):  
Escherichia Coli ◽  
Raw Milk ◽  
Most Probable Number ◽  
Prolonged Storage ◽  
Escherichia Coli O157 ◽  
Probable Number ◽  
E Coli ◽  
Aging Period ◽  
Gouda Cheese ◽  
Cheese Products

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.

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