ICMSF methods studies. XIII. An international comparative study of the MPN procedure and the Anderson–Baird–Parker direct plating method for the enumeration of Escherichia coli biotype I in raw meats

1979 ◽  
Vol 25 (11) ◽  
pp. 1321-1327 ◽  
Author(s):  
M. K. Rayman ◽  
G. A. Jarvis ◽  
C. M. Davidson ◽  
S. Long ◽  
J. M. Allen ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Comparative Study ◽  
Most Probable Number ◽  
Probable Number ◽  
Direct Plating ◽  
E Coli ◽  
Plating Method ◽  
International Comparative ◽  
Lower Variability ◽  
Raw Meats

The most probable number (MPN) and a direct membrane-plating (DP) method were compared for enumeration of Escherichia coli biotype I in raw meats by 11 laboratories. The DP method yielded higher counts of E. coli than the MPN method for frozen samples but neither method consistently gave higher counts for non-frozen samples. The DP method was less variable and gave a higher rate of detection of low numbers of E. coli in frozen samples. Despite the inability of the DP method to enumerate E. coli biotype II and intermediate types, which comprise only 3–5% of the Escherichia strains (Ewing 1972), the method is preferable to the MPN method for enumerating E. coli in raw meats because of its lower variability, better recovery from frozen samples, rapidity, decreased requirement for media, and decreased costs for analysts' time.

The Anderson–Baird-Parker direct plating method versus the most probable number procedure for enumerating Escherichia coli in meats

1981 ◽  
Vol 27 (1) ◽  
pp. 147-149 ◽  
Author(s):  
M. K. Rayman ◽  
B. Aris
Keyword(s):  
Escherichia Coli ◽  
North American ◽  
Most Probable Number ◽  
Probable Number ◽  
Direct Plating ◽  
E Coli ◽  
Membrane Filters ◽  
The North ◽  
Plating Method

Comparison of the Anderson–Baird-Parker direct plating method (DP) and the North American most probable number procedure (MPN) for enumerating Escherichia coli in frozen meats revealed that the DP method is more precise and yields higher counts of E. coli than the MPN procedure. Any of three brands of membrane filters tested was suitable for use in the DP method.

A Direct Plating Method for Estimating Populations of Escherichia coli O157 in Bovine Manure and Manure-Based Materials†

2008 ◽  
Vol 71 (11) ◽  
pp. 2233-2238 ◽  
Author(s):  
ELAINE D. BERRY ◽  
JAMES E. WELLS
Keyword(s):  
Escherichia Coli ◽  
Soil Amendments ◽  
Most Probable Number ◽  
Escherichia Coli O157 ◽  
Surface Material ◽  
Potassium Tellurite ◽  
Probable Number ◽  
Direct Plating ◽  
E Coli ◽  
Plating Method

Escherichia coli O157:H7 outbreaks associated with produce consumption have brought attention to livestock manures and manure-based soil amendments as potential sources of pathogens for the contamination of these crops. Procedures for enumeration of E. coli O157:H7 are needed to assess the risks of transmission from these manures and their by-products. A direct plating method employing spiral plating onto CHROMagar O157 was investigated for enumeration of E. coli O157:H7 in feedlot surface material, aged bovine manure, bovine manure compost, and manure-amended soil. In studies utilizing samples spiked with a five-strain cocktail of E. coli O157:H7 at levels ranging from 102 to 105 CFU/g of sample, there were strong correlations between the observed and predicted levels of this pathogen. Although the addition of 2.5 mg/liter potassium tellurite and 5 mg/liter novobiocin made the medium more restrictive, these amendments enhanced the ability to identify and enumerate E. coli O157:H7 in feedlot surface material, which contained a higher proportion of fresh feces than did the other three sample types and therefore higher levels of interfering bacterial microflora. The spiral plating method was further assessed to determine its ability to enumerate E. coli O157:H7 in naturally contaminated feedlot surface material. Comparison of E. coli O157:H7 counts in feedlot surface material obtained by the spiral plating method and a most probable number technique were well correlated. We conclude that direct spiral plating onto CHROMagar O157 is effective for estimating E. coli O157: H7 levels in a variety of manures and manure-containing sample types to a lower detection limit of 200 CFU/g. The method has application for determining E. coli O157:H7 concentrations in manures and composts before their sale and use as soil amendments and for measuring the effectiveness of manure treatment processes to reduce or inactivate this pathogen.

scholarly journals Comparison of the Compact Dry EC with the Most Probable Number Method (AOAC Official Method 966.24) for Enumeration of Escherichia coli and Coliform Bacteria in Raw Meats: Performance-Tested MethodSM 110402

2006 ◽  
Vol 89 (1) ◽  
pp. 100-114 ◽  
Author(s):  
Hidemasa Kodaka ◽  
Shingo Mizuochi ◽  
Hajime Teramura ◽  
Tadanobu Nirazuka ◽  
David Goins ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Water Soluble ◽  
Test Method ◽  
Gelling Agent ◽  
Coliform Bacteria ◽  
Most Probable Number ◽  
Official Method ◽  
Probable Number ◽  
E Coli ◽  
Raw Meats

Abstract Compact Dry E. coli/Coliform Count (EC) is a ready-to-use test method for the enumeration of Escherichia coli and coliform bacteria in food. The plates are presterilized and contain culture medium and a cold water-soluble gelling agent. The medium should be rehydrated with 1 mL diluted sample inoculated onto the center of the self-diffusible medium, allowing the solution to diffuse by capillary action. The plate can be incubated at 35C for 2024 h and the colonies counted without any further working steps. The Compact Dry EC medium plates were validated as an analysis tool for determining colony-forming units (CFU) of E. coli and coliform bacteria from a variety of raw meats using 5 different types of raw meats. The performance tests were conducted at 35C. In all studies performed, no apparent differences were observed between the Compact Dry ECmethod and theAOAC Official Method 966.24 results. For the accuracy claim (n = 75), a correlation factor of r2 = 0.93 (E. coli) and r2 = 0.93 (coliform bacteria) could be assigned, as stated in the application for Performance-Tested MethodSM.

ColiComplete® Substrate-Supporting Disc Method for Confirmed Detection of Total Coliforms and Escherichia coli in All Foods: Comparative Study

1994 ◽  
Vol 77 (1) ◽  
pp. 58-63 ◽  
Author(s):  
Philip T Feldsine ◽  
Maria T Falbo-Nelson ◽  
David L Hustead
Keyword(s):  
Escherichia Coli ◽  
Comparative Study ◽  
Total Coliform ◽  
Coliform Bacteria ◽  
Most Probable Number ◽  
Probable Number ◽  
Total Coliforms ◽  
E Coli ◽  
Total Coliform Bacteria ◽  
Better Than

Abstract The ColiComplete® substrate-supporting disc (SSD) method for simultaneous confirmed total coliform count and Escherichia coli determination in all foods was compared with the AOAC most probable number (MPN) methods 966.23 and 966.24. In this comparative study, 20 water and food types were analyzed; 7 of these foods were naturally contaminated with coliform bacteria, 6 food types were naturally contaminated with E. coli, and the remaining foods were inoculated with coliform bacteria and/or E. coli. Data were analyzed separately for total coliform bacteria and for E. coli. Mean log MPN counts were determined by the SSD method and the appropriate AOAC MPN procedure. Results were then analyzed for mean log MPN differences and variance, according to methods described by AOAC INTERNATIONAL Results for both total conforms and E. coli indicate that the SSD method is equivalent to or better than AOAC MPN methods 966.23 and 966.24.

Development and Validation of a Most-Probable-Number–Immunomagnetic Separation Methodology of Enumerating Escherichia coli O157 in Cattle Feces

2007 ◽  
Vol 70 (5) ◽  
pp. 1072-1075 ◽  
Author(s):  
T. P. STEPHENS ◽  
G. H. LONERAGAN ◽  
W. E. CHANEY ◽  
L. A. BRANHAM ◽  
M. M. BRASHEARS
Keyword(s):  
Escherichia Coli ◽  
Immunomagnetic Separation ◽  
Feedlot Cattle ◽  
Most Probable Number ◽  
Escherichia Coli O157 ◽  
Probable Number ◽  
Direct Plating ◽  
Fecal Samples ◽  
E Coli ◽  
The Difference

A method to validate enumeration of Escherichia coli O157 in fecal samples from feedlot cattle was developed in these studies. Due to background flora, bovine fecal sample enumeration cannot be performed by simple direct plating techniques. Known quantities of E. coli O157:H7 were inoculated into feces, and populations were determined by direct plating of the cocktail (studies 1, 2, and 3) and manure and cocktail (studies 4 and 5) mixtures and compared with a most-probable-number (MPN)–immunomagnetic separation (IMS) method. The three-tube MPN combined preenrichment in gram-negative broth with confirmation using IMS. Five separate enumeration studies (study 1, sterile feces inoculated with 102 E. coli O157:H7 per g; study 2, nonsterile feces inoculated with 103 E. coli O157:H7 per g; study 3, nonsterile feces inoculated with 101 E. coli O157:H7 per g; study 4, sterile feces inoculated with 104 streptomycin-resistant E. coli O157:H7 per g; and study 5, sterile feces inoculated with 102 streptomycin-resistant E. coli O157:H7 per g) were conducted. These studies were performed to determine the precision, accuracy, and specificity at low and high levels of pathogen contamination in feces, using direct plating compared with the MPN-IMS methodology tested. There was an overall difference (P < 0.01) between direct plating and MPN-IMS methodologies, but this difference was biologically negligible due to the difference in least-squares means (0.29 ± 0.10) being so low. The direct plating and MPN-IMS methods were correlated (r = 0.93). These results suggest that using the MPN-IMS procedures is an effective method of estimating E. coli O157 populations in naturally infected bovine fecal samples.

scholarly journals Dry Rehydratable Film Method for Enumerating Confirmed Escherichia coli in Poultry, Meats, and Seafood: Collaborative Study

1999 ◽  
Vol 82 (1) ◽  
pp. 73-78 ◽  
Author(s):  
Vidhya Gangar ◽  
Michael S Curiale ◽  
Kathryn Lindberg ◽  
Sonya Gambrel-Lenarz ◽  
E Adamson ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Collaborative Study ◽  
Most Probable Number ◽  
Probable Number ◽  
E Coli ◽  
Fish Samples ◽  
Plating Method ◽  
Medium Level ◽  
Repeatability And Reproducibility ◽  
Film Method

Abstract A rehydratable dry-film plating method for Escherichia coli, the Petrifilm E. coli/Coliform (EC) Count Plate in foods, has been compared with the AOAC INTERNATIONAL most probable number (MPN) method. Eleven laboratories participated in the collaborative study. Three E. coli levels in 8 samples each of frozen raw ground turkey, frozen raw ground beef, and frozen cooked fish were tested in duplicate. Mean log counts for the Petri film plate procedure were not significantly different from those for the MPN procedure for cooked fish samples inoculated with low or high inocula levels, for samples of raw turkey inoculated at medium level, and for beef inoculated at low, medium, and high levels. Repeatability and reproducibility vari ances of the Petrifilm EC Plate method recorded at 24 h were as good as or better than those of the MPN method. The dry rehydratable film method for enumerating confirmed E. coli in poultry, meats, and seafood has been adopted first action by AOAC INTERNATIONAL.

Use of the Chromogenic Substrate 5-Bromo-4-Chloro-3-Indolyl-B-D-Glucuronide (X-GLUC) for Enumerating Escherichia coli in 24 H from Ground Beef

1990 ◽  
Vol 53 (6) ◽  
pp. 508-510 ◽  
Author(s):  
LAWRENCE RESTAINO ◽  
ELON W. FRAMPTON ◽  
RICHARD H. LYON
Keyword(s):  
Escherichia Coli ◽  
Ground Beef ◽  
Most Probable Number ◽  
Chromogenic Substrate ◽  
Probable Number ◽  
E Coli ◽  
Glucuronidase Activity ◽  
Significant Difference ◽  
Plating Method ◽  
Positive Linear Correlation

A 24-h direct plating method for Escherichia coli using the chromogenic substrate 5-bromo-4-chloro-3-indolyl-B-D-glucuronide (X-GLUC) incorporated into a Peptone-tergitol agar base (PTX) was compared with the standard 3-tube Most Probable Number (MPN) method on 50 naturally contaminated ground beef samples. A paired-comparisons t-test showed no significant difference between the two methods. A positive linear correlation between the two methods was observed over the entire range of values. Ninety-seven percent of the positive colonies (blue colonies) on PTX agar were indentified as E. coli, whereas no atypical colonies (nonblue) were characterized as such. Thus, a simple and reliable enumeration of E. coli can be made within 24 h using the X-GLUC substrate in a selective agar as an indicator of B-glucuronidase activity.

scholarly journals Comparison of Fecal Coliform Agar and Violet Red Bile Lactose Agar for Fecal Coliform Enumeration in Foods

2002 ◽  
Vol 68 (4) ◽  
pp. 1631-1638 ◽  
Author(s):  
A. Leclercq ◽  
C. Wanegue ◽  
P. Baylac
Keyword(s):  
Escherichia Coli ◽  
Fecal Coliform ◽  
Food Samples ◽  
Fecal Coliforms ◽  
Predictive Values ◽  
Direct Plating ◽  
Hafnia Alvei ◽  
E Coli ◽  
Plating Method ◽  
Mashed Potatoes

ABSTRACT A 24-h direct plating method for fecal coliform enumeration with a resuscitation step (preincubation for 2 h at 37 ± 1°C and transfer to 44 ± 1°C for 22 h) using fecal coliform agar (FCA) was compared with the 24-h standardized violet red bile lactose agar (VRBL) method. FCA and VRBL have equivalent specificities and sensitivities, except for lactose-positive non-fecal coliforms such as Hafnia alvei, which could form typical colonies on FCA and VRBL. Recovery of cold-stressed Escherichia coli in mashed potatoes on FCA was about 1 log unit lower than that with VRBL. When the FCA method was compared with standard VRBL for enumeration of fecal coliforms, based on counting carried out on 170 different food samples, results were not significantly different (P > 0.05). Based on 203 typical identified colonies selected as found on VRBL and FCA, the latter medium appears to allow the enumeration of more true fecal coliforms and has higher performance in certain ways (specificity, sensitivity, and negative and positive predictive values) than VRBL. Most colonies clearly identified on both media were E. coli and H. alvei, a non-fecal coliform. Therefore, the replacement of fecal coliform enumeration by E. coli enumeration to estimate food sanitary quality should be recommended.

scholarly journals Lessons from a large outbreak of Escherichia coli O157[ratio ]H7 infections: insights into the infectious dose and method of widespread contamination of hamburger patties

1999 ◽  
Vol 122 (2) ◽  
pp. 185-192 ◽  
Author(s):  
J. TUTTLE ◽  
T. GOMEZ ◽  
M. P. DOYLE ◽  
J. G. WELLS ◽  
T. ZHAO ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Ground Beef ◽  
Most Probable Number ◽  
Strong Argument ◽  
Probable Number ◽  
Infectious Dose ◽  
E Coli ◽  
Beef Patties ◽  
Large Outbreak ◽  
Microbiological Testing

Between November 1992 and February 1993, a large outbreak of Escherichia coli O157[ratio ]H7 infections occurred in the western USA and was associated with eating ground beef patties at restaurants of one fast-food chain. Restaurants that were epidemiologically linked with cases served patties produced on two consecutive dates; cultures of recalled ground beef patties produced on those dates yielded E. coli O157[ratio ]H7 strains indistinguishable from those isolated from patients, confirming the vehicle of illness. Seventy-six ground beef patty samples were cultured quantitatively for E. coli O157[ratio ]H7. The median most probable number of organisms was 1·5 per gram (range, <0·3–15) or 67·5 organisms per patty (range, <13·5–675). Correlation of the presence of E. coli O157[ratio ]H7 with other bacterial indicators yielded a significant association between coliform count and the presence of E. coli O157[ratio ]H7 (P=0·04). A meat traceback to investigate possible sources of contamination revealed cattle were probably initially colonized with E. coli O157[ratio ]H7, and that their slaughter caused surface contamination of meat, which once combined with meat from other sources, resulted in a large number of contaminated ground beef patties. Microbiological testing of meat from lots consumed by persons who became ill was suggestive of an infectious dose for E. coli O157[ratio ]H7 of fewer than 700 organisms. These findings present a strong argument for enforcing zero tolerance for this organism in processed food and for markedly decreasing contamination of raw ground beef. Process controls that incorporate microbiological testing of meat may assist these efforts.

scholarly journals Mechanically Ventilated Broiler Sheds: a Possible Source of Aerosolized Salmonella, Campylobacter, and Escherichia coli

2009 ◽  
Vol 75 (23) ◽  
pp. 7417-7425 ◽  
Author(s):  
H. N. Chinivasagam ◽  
T. Tran ◽  
L. Maddock ◽  
A. Gale ◽  
P. J. Blackall
Keyword(s):  
Escherichia Coli ◽  
Most Probable Number ◽  
Production Cycle ◽  
Airborne Bacteria ◽  
Probable Number ◽  
Mechanically Ventilated ◽  
The Public ◽  
E Coli ◽  
Poultry Farms ◽  
Low Levels

ABSTRACT This study assessed the levels of two key pathogens, Salmonella and Campylobacter, along with the indicator organism Escherichia coli in aerosols within and outside poultry sheds. The study ranged over a 3-year period on four poultry farms and consisted of six trials across the boiler production cycle of around 55 days. Weekly testing of litter and aerosols was carried out through the cycle. A key point that emerged is that the levels of airborne bacteria are linked to the levels of these bacteria in litter. This hypothesis was demonstrated by E. coli. The typical levels of E. coli in litter were ∼108 CFU g−1 and, as a consequence, were in the range of 102 to 104 CFU m−3 in aerosols, both inside and outside the shed. The external levels were always lower than the internal levels. Salmonella was only present intermittently in litter and at lower levels (103 to 105 most probable number [MPN] g−1) and consequently present only intermittently and at low levels in air inside (range of 0.65 to 4.4 MPN m−3) and once outside (2.3 MPN m−3). The Salmonella serovars isolated in litter were generally also isolated from aerosols and dust, with the Salmonella serovars Chester and Sofia being the dominant serovars across these interfaces. Campylobacter was detected late in the production cycle, in litter at levels of around 107 MPN g−1. Campylobacter was detected only once inside the shed and then at low levels of 2.2 MPN m−3. Thus, the public health risk from these organisms in poultry environments via the aerosol pathway is minimal.

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