MicroVal Study 2017LR76 - Betty Blocks€¦ · The 3M Petrifilm Rapid E. coli / Coliform Count...

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MicroVal Study 2017LR76

Validation of the 3M™ Petrifilm™ ™ Rapid E. coli / Coliform Count Plate for the Enumeration of E. coli and Coliform in a Broad Range of Foods

following ISO 16140-2:2016 Guidelines: Method Comparison Study Report

Version 2: MLS + ILS December 14th, 2018

Contact Details: Q Laboratories 1400 Harrison Ave. Cincinnati, OH 45214 Tel: 513-471-1300 www.qlaboratories.com

Report issued and authorized by: Q Laboratories Benjamin Bastin, Microbiology R&D Supervisor

http://www.qlaboratories.com/

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The MCS report is prepared in accordance with ISO 16140-1:2016 [1] and

ISO 16140-2:2016 [2] and the most recent version of the MicroVal Technical

Committee for interpretation on ISO 16140-2, version 1.0.

Company: 3M Food Safety

Expert Laboratory: Q Laboratories

Method/Kit name: 3M™ Petrifilm™ Rapid E. coli / Coliform Count Plate

Validation standard: ISO 16140-2:2016 Microbiology of food chain – Method

Validation – Part 2: Protocol for the validation of alternative (proprietary) methods

against a reference method.

Reference methods:

1. ISO 4832:2006, Microbiology of food and animal feeding stuffs – Horizontal

method for the enumeration of coliforms – Colony-count technique

2. ISO 16649-2:2001, Microbiology of food and animal feeding stuffs –

Horizontal method for the enumeration of β-glucuronidase-positive

Escherichia coli – Part 2: Colony count technique.at 44 °C using 5-bromo-4-

chloro-3-indolyl β-D-glucuronide

Scope of validation: Broad Range of Foods and Select Environmental Surfaces.

Food Categories tested: Meat Products, Dairy and Egg Products, Fish and Seafood

Products, Vegetable Products, Ready to Eat and Ready to Reheat, Animal Feed and

Samples from the Industrial Production Environment

Certification organization: Lloyd's Register

List of abbreviations

- AL Acceptability Limit

- AP Accuracy Profile

- Art. Cont. Artificial contamination

- CFU Colony Forming Units

- CL Confidence Limit (usually 95%)

- EL Expert Laboratory

- �̅� Average difference

- g Gram

- h Hour

- ILS Interlaboratory Study

- Incl/Excl Inclusivity and Exclusivity

- MCS Method Comparison Study

- min minute

- ml Millilitre

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- MR (MicroVal) Method Reviewer

- MVTC MicroVal Technical Committee

- n number of samples

- na not applicable

- RT Relative Trueness

- SD Standard Deviation of Differences

- 10-1 dilution 10-fold dilution of original food

- 10-2 dilution 100-fold dilution of original food

- VRBG/L Violet Red Bile Glucose Agar with Lactose

- TBX Tryptone Bile X-Glucuronide Medium

- TNTC Too numerous to count

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Table of Contents

1 Introduction 5

2 Method Protocols 6

2.1 Reference Method – ISO 4832 6

2.2 Reference Method – ISO 16649-2:2001 6

2.3 Alternative method 6

3 Methods Comparison Study 6

3.1 Sample Preparation 6

3.2 Trueness Study 7

3.2.1 Calculations and Interpretation 9

3.2.2 Conclusion of the Truness Study 28

3.3 Accuracy Profile Study 28

3.3.1 Calculation and Data Interpretation 30

3.3.2 Conclusion of the Accuracy Profile Study 31

3.4 Inclusivity and Inclusivity and Exclusivity Study 70

4 Interlaboratory Study 74

4.1 Organization 74

4.2 Calculations and interpretation of data 75

5 References 85

6 ANNEX A: Flow Diagram of the ISO 4832 Reference Method 86

7 ANNEX B: Flow Diagram of the ISO 16649-2:2001 Reference Method 87

8 ANNEX C: Flow Diagram of the 3M Petrifilm Rapid E. coli and Coliform Count Plate Alternative Method 88

9 ANNEX D: Kit Insert for 3M Petrifilm Rapid E. coli / Coliform Count Plate 88

See the attached document. 88

10 ANNEX E: Summary of Artificial Contamination 89

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1 Introduction

The 3M™ Petrifilm™Rapid E. coli / Coliform Count Plate was validated according to ISO 16140-2:2016 for the enumeration of coliforms and E. coli in a broad range of foods and environmental surfaces. All testing was conducted by Q Laboratories, Inc.

Below is a summary of the 3M Petrifilm Rapid E. coli / Coliform Count Plate Method.

For each food type, 10 g test portions were diluted at a 1:10 ratio using the appropriate diluent. Each environmental sponge was diluted with 90 mL of diluent. Paired 10 g test portions were evaluated for the alternative and reference methods for all food types in the scope.

1.1. After dilution, 1.0 mL of sample was dispensed onto the 3MPetrifilm Rapid E. coli / Coliform Count Plate plate. Plates were incubated at 30 ± 1°C for dairy products and 37 ± 1°C and 42 ± 1°C for all other matrices. All colonies were enumerated at 18 hours.

1.2. For the purposes of the MicroVal validation, up to 5 atypical coliform colonies were confirmed through transfer into tubes of Brilliant Green Lactose Bile (BGLB) Broth and incubating at 24 ± 2 hours at 37 ± 1 ºC. Production of gas indicated the presence of coliform.

Reference methods: ISO 4832:2006 [3] and ISO 16649-2:2001 [4]. See Annex A-E for flow charts and kit inserts of the methods.

Scope of the validation study is: Broad range of food matrices and select environmental surfaces.

Samples were prepared in accordance with ISO 6887: parts 1 – 5. [5]

Categories included:

− Meat Products (Combined category raw/RTC meats and poultry)

− Dairy and Egg Products

− Fish and Seafood Products

− Vegetable Products

− Ready to Eat and Ready to Reheat (Combined category RTE/RTRH meats and poultry)

− Animal Feed

− Samples from the Industrial Production Environment

Criteria evaluated during the study were:

− Method Comparison Study (MCS)

• Relative Trueness study

• Relative Accuracy study

• Inclusivity and exclusivity study

− Interlaboratory Study (ILS)

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2 Method Protocols

The MCS was carried out using 10 gram portions of sample material for all food matrices and environmental sponges for all samples from the industrial production environment. In both the trueness and accuracy studies, 3M Petrifilm Rapid E. coli / Coliform Count Plate was evaluated at 18 hours of incubation. A paired design was utilized for all foods, as the reference and the alternative

method shared the same diluent.

2.1 Reference Method – ISO 4832

See the flow diagram in Annex A.

Sample preparations used in the reference method were done according to ISO

6887-series.

2.2 Reference Method – ISO 16649-2:2001

See the flow diagram in Annex B.

Sample preparations used in the reference method were done according to ISO 6887-series.

2.3 Alternative method

See the flow diagram of the alternative method in Annex C and the 3M Petrifilm

Rapid E. coli / Coliform Count Plate kit insert in Annex D for a detailed description of

the method.

The 3M Petrifilm Rapid E. coli / Coliform Count Plate test is a selective and differential sample-ready-culture medium system which contains proprietary nutrients, a cold-water-soluble gelling agent, 5-bromo-4-chloro-3-indoyl-D-glucuronide (BCIG), an indicator of glucuronidase activity, and a tetrazolium indicator that facilitates colony enumneration.

The food samples were prepared using 10 g test portions or an environmental surface sponge diluted with 90 mL of the appropriate diluent. Colonies were enumerated after 18 hours of incubation at specified temperatures (37 ± 1oC for coliform and E. coli; 42 ± 1oC for E. coli); (30 ± 1°C for dairy coliform and E. coli).

3 Methods Comparison Study

3.1 Sample Preparation

The Method Comparison Study was carried out using 10 g test portions or sponges for environmental industrial production samples. The samples were prepared for analysis and diluted in accordance with ISO 6887 (all parts). A 1:10 dilution was performed for all food types. All analysis were conducted using paired samples

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3.2 Trueness Study

The selected types per category tested are listed in Table 1.

In accordance with ISO 16140-2, for each category, a minimum of 15 samples were tested. Within each type, a minimum of 5 samples (producing interpretable results) that cover a representative variety of products were evaluated. All samples covered a wide range of contamination that could be observed in the food items (i.e. 10 CFU/g to 105 CFU/g).

Table 1. List of Categories and Types of Items to be Tested within the Trueness Study

Category Food Type Number of

Samples1

Test Portion

Size

Meat Products

(Combined

category raw/RTC

meats and poultry)

Raw Meat 5 10 g

Breaded Meats

(i.e. Frozen Wings) 5 10 g

Cured and Smoked Products (raw) 5 10 g

Dairy and Egg

Products

Pasteurized Dairy Products 5 10 g

Raw Dairy Products 5 10 g

Desserts and Egg Products 5 10 g

Fish and Seafood Products

Raw Products 5 10 g

Marinated Smoked Items 5 10 g

Cooked Products 5 10 g

Vegetable Products

Raw Vegetables 5 10 g

Pre-Cooked Vegetables 5 10 g

Processed Vegetables 5 10 g

Ready to Eat and Ready to Reheat

Cold Ready Meals 5 10 g

Pasta 5 10 g

Mayonnaise based deli salads with

processed ingredients 5 10 g

Animal Feed

Finished Products for Domestic 5 10 g

Finished Products for Livestock 5 10 g

Ingredients for domestic/livestock 5 10 g

Samples from the

Industrial

Production

Environment2

Surface Samples 15 Environmental

Sponge

1Total number of samples = minimum of 15 paired samples/category; 2 Limited claims for surface samples only

Samples were evaluated for natural contamination of the target organisms. Thirty (30%) of total samples evaluted were naturally contaminated with coliform and E. coli. For positive samples used in the evaluation, 63% of samples were obtained from natural contamination. The following spiking protocol indicates procedures to inoculate a wide range of food items. Not all spiking protocols were required for the validation.

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1.1. For high water activity products, a liquid inoculum was heated at 50 °C for 10 minutes prior to inoculation.

1.2. For low water activity products, a lyophilized culture was used. 1.3. The stressed cultures was used to inoculate the relevant food matrix and

would be analyzed immediately. The level of injury was evaluated by plating on a non-selective agar (Tryptic Soy Agar) and a selective agar (TBX for Escherichia coli and VRBL for Coliform). It was proposed to obtain between a 50% (0.3log) and a 70% (0.5log) injured cells.

The following seeding protocol indicates procedures to inoculate a wide range of food items. Not all seeding protocols were required for the validation.

1.1. Overnight cultures of Escherichia coli and coliform were inoculated at the target contamination level into the food matrix and then stored under relevant conditions for the food matrix being analyzed prior to analysis:

1.2. For refrigerated products, samples were stored at 5 °C for 48 to 72 hours. 1.3. For frozen foods, samples were stored at -20 °C for a minimum of 2 weeks. 1.4. For shelf stable products, samples were stored at room temperature (22-25

°C) for a minimum of 2 weeks.

Table 2 provides a list of strains that were used for artificial inoculations. These cultures originated from comparable sample types as the ones to be inoculated.

Table 2. List of Proposed Inoculation Organisms for the Trueness Study

Organisms Source Origin

Citrobacter freundii QL1 100813-2A Sliced deli turkey meat

Cronobacter condimenti DSM2 27966 Infant Formula

Enterobacter amnigenus ATCC3 51816 Milk, Minnesota

Hafnia alvei ATCC 51815 Milk, Minnesota

Klebsiella pneumonia QL 11007-7 Raw hamburger

Rahnella aqualitis ATCC 55046 Soil

Serratia marcescens ATCC 8100 Milk, Cork Ireland

Escherichia coli QL 16160-4 Chicken Nuggets

Escherichia coli QL 11010-2 Bottled Water

Escherichia coli QL 16154-7 Flour

Escherichia coli QL 16154-2 Raw Ground Beef

Escherichia coli QL16160-2 Chicken Carcass Rinsate

Escherichia coli QL 16160-10 Raw Pork

Escherichia coli QL 16160-2 Mixed Frozen Vegetables

Escherichia coli QL 16154-3 Beef Trim

Escherichia coli QL 16154-13 Potable Water

Escherichia coli QL 16154-2 Raw Ground Beef

Escherichia coli QL 16154-7 Flour 1 QL-Q Laboratories Culture Collection , 2 DSM-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, 3 ATCC-American Type Culture Collection The inoculation of samples was at the range usually associated with the test organisms and within the capabilities of the test methods. Enumeration methods covered the range 10 CFU/g to 105 CFU/g.

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3.2.1 Calculations and Interpretation

All results were tabulated, calculated and interpreted according to ISO 16140-2. The obtained results for each food item per category and all food items in all categories were plotted. The line of identity was drawn (y=x) . The data obtained was analzyed using the scatter plot and the line of identity. The figures below display a summary of the data. The raw data from the relative trueness study are presented in Annex F.

Figure 1A: Meat Products – Coliform 37 ± 1°C

Figure 1B: Dairy and Egg Products – Coliform 30 ± 1°C & 37 ± 1°C

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Figure 1C: Fish and Seafood Products – Coliform 37 ± 1°C

Figure 1D: Vegetable Products – Coliform 37 ± 1°C

Figure 1E: Ready to Eat and Ready to Reheat Products – Coliform 37 ± 1°C

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Figure 1F: Animal Feed Products – Coliform 37 ± 1°C

Figure 1G: Surface Samples – Coliform 37 ± 1°C

Figure 1H: All Categories – Coliform 30 ± 1°C & 37 ± 1°C1

130°C for dairy products and 37°C for all other matrices

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Figure 1I: Meat Products – E. coli 37 ± 1°C

Figure 1J: Meat Products – E. coli 42 ± 1°C

Figure 1K: Dairy and Egg Products – E. coli 30 ± 1°C & 37 ± 1°C

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Figure 1L: Dairy and Egg Products – E. coli 30 ± 1°C & 42 ± 1°C

Figure 1M: Fish and Seafood Products – E. coli 37 ± 1°C

Figure 1N: Fish and Seafood Products – E. coli 42 ± 1°C

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Figure 1O: Vegetable Products – E. coli 37 ± 1°C

Figure 1P: Vegetable Products – E. coli 42 ± 1°C

Figure 1Q: Ready to Eat and Ready to Reheat Products – E. coli 37 ± 1°C

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Figure 1R: Ready to Eat and Ready to Reheat Products – E. coli 42 ± 1°C

Figure 1S: Animal Feed Products – E. coli 37 ± 1°C

Figure 1T: Animal Feed Products – E. coli 42 ± 1°C

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Figure 1U: Surface Samples – E. coli 37 ± 1°C

Figure 1V: Surface Samples – E. coli 42 ± 1°C

Figure 1W: All Categories – E. coli 30 ± 1°C & 37 ± 1°C1


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Figure 1X: All Categories – E. coli 30 ± 1°C & 42 ± 1°C1


According to ISO 16140-2:2012, section 6.1.2.3, the results of the scatter plots and the line of identity were interpreted based on a visual observation on the amount of bias and extreme results. The data appeared to be acceptable per category and as a whole. The obtained results were analyzed using the Bland-Altman method as detailed in section 6.1.2.3 in ISO 16140-2. Table 3. Summary of the Calculated Differences and Confidence Limits

Organism Food

Category Temperature n

95%

Lower

Limit

95%

Upper

Limit

Coliform

Meat

Products 37 ± 1°C 15 0.0238 0.0355 -0.0523 0.0999

Dairy and

Egg

Products

30 ± 1°C &

37 ± 1°C 15 0.0463 0.0528 -0.0669 0.1595

Fish and

Seafood

Products

37 ± 1°C 15 0.0113 0.0371 -0.0683 0.0909

Vegetable

Products 37 ± 1°C 15 0.0126 0.0504 -0.0955 0.1206

Ready to

Eat and

Ready to

Reheat

37 ± 1°C 15 0.0051 0.0342 -0.0682 0.0784

Animal Feed

Products 37 ± 1°C 15

-0.0031

0.0238 -0.0540 0.0479

Surface

Samples 37 ± 1°C 15

-0.0031

0.0348 -0.0777 0.0715

All

Categories

30 ± 1°C &

37 ± 1°C 105 0.0133 0.0416 -0.0692 0.0957

D Ds

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Organism Food

Category Temperature n

95% Lower Limit

95% Upper Limit

E. coli

Meat

Products

37 ± 1°C 15 -

0.0714 0.0446 -0.1670 0.0242

42 ± 1°C 15 0.0001 0.0927 -0.1987 0.1990

Dairy and

Egg

Products

30 ± 1°C &

37 ± 1°C 15

-

0.0126 0.0649 -0.1518 0.1267

30 ± 1°C &

42 ± 1°C 15

-

0.0073 0.0623 -0.1409 0.1263

Fish and

Seafood

Products

37 ± 1°C 15 -

0.1019 0.1284 -0.3774 0.1736

42 ± 1°C 15 -

0.0365 0.0892 -0.2277 0.1547

Vegetable

Products

37 ± 1°C 15 -

0.0834 0.1324 -0.3673 0.2005

42 ± 1°C 15 -

0.0359 0.1761 -0.4137 0.3419

Ready to

Eat and

Ready to

Reheat

37 ± 1°C 15 -

0.0831 0.1204 -0.3412 0.1751

42 ± 1°C 15 -

0.0305 0.0851 -0.2131 0.1521

Animal Feed

Products

37 ± 1°C 15 -

0.0726 0.1220 -0.3344 0.1892

42 ± 1°C 15 0.0193 0.1451 -0.2919 0.3306

Surface

Samples

37 ± 1°C 15 -

0.1236 0.1392 -0.4222 0.1750

42 ± 1°C 15 -

0.0001 0.1731 -0.3715 0.3712

All

Categories

30 ± 1°C &

37 ± 1°C 105

-

0.0784 0.1140 -0.3045 0.1477

30 ± 1°C &

42 ± 1°C 105

-

0.0053 0.1244 -0.2520 0.2413

The results of the difference and scatter plot were interpreted, based on a visual observation on the amount of bias and extreme results. It was expected that not more than 1 in 20 data values will lie outside the confidence limits. Any disagreements with the expectation were recorded.

The symbols used ensure the different types of product in each category are clearly identified. The average of each pair of data values and the difference between these values were calculated for each sample and for each category.

D Ds

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Figure 2A: Bland-Altman for Meat Products – Coliform 37 ± 1°C

Figure 2B: Bland-Altman for Dairy and Egg Products – Coliform 30 ± 1°C & 37 ± 1°C

Figure 2C: Bland-Altman for Fish and Seafood Products – Coliform 37 ± 1°C

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Figure 2D: Bland-Altman for Vegetable Products – Coliform 37 ± 1°C

Figure 2E: Bland-Altman for Ready to Eat and Ready to Reheat Products – Coliform 37 ± 1°C

Figure 2F: Bland-Altman for Animal Feed Products – Coliform 37 ± 1°C

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Figure 2G: Bland-Altman for Surface Samples – Coliform 37 ± 1°C

Figure 2H: Bland-Altman for All Categories – Coliform 30 ± 1°C & 37 ± 1°C1


Figure 2I: Bland-Altman for Meat Products – E. coli 37 ± 1°C

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Figure 2J: Bland-Altman for Meat Products – E. coli 42 ± 1°C

Figure 2K: Bland-Altman for Dairy and Egg Products – E. coli 30 ± 1°C & 37 ± 1°C

Figure 2L: Bland-Altman for Dairy and Egg Products – E. coli 30 ± 1°C & 42 ± 1°C

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Figure 2M: Bland-Altman for Fish and Seafood Products – E. coli 37 ± 1°C

Figure 2N: Bland-Altman for Fish and Seafood Products – E. coli 42 ± 1°C

Figure 2O: Bland-Altman for Vegetable Products – E. coli 37 ± 1°C

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Figure 2P: Bland-Altman for Vegetable Products – E. coli 42 ± 1°C

Figure 2Q: Bland-Altman for Ready to Eat and Ready to Reheat Products – E. coli 37 ± 1°C

Figure 2R: Bland-Altman for Ready to Eat and Ready to Reheat Products – E. coli 42 ± 1°C

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Figure 2S: Bland-Altman for Animal Feed Products – E. coli 37 ± 1°C

Figure 2T: Bland-Altman for Animal Feed Products – E. coli 42 ± 1°C

Figure 2U: Bland-Altman for Surface Samples – E. coli 37 ± 1°C

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Figure 2V: Bland-Altman for Surface Samples – E. coli 42 ± 1°C

Figure 2W: Bland-Altman for All Categories – E. coli 30 ± 1°C & 37 ± 1°C1


Figure 2X: Bland-Altman for All Categories – E. coli 30 ± 1°C & 42 ± 1°C1


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For all Categories, there were five out of 105 food items which lie outside the CLs for Coliform, ten out of 105 food items for E. coli incubated at 37 ± 1°C and thirteen out of 105 food items for E. coli incubated at 42 ± 1°C.

The 28 total food items which are outside of the CLs are shown in Table 4. There were no identifiable trends in these data as they covered both target organisms at all three temperatures in 7 different food types and being associated with both natural and artificial contamination. For all of the results that were outside of the confidence limits, there were zero results that were greater than a 0.5 Log difference indicating that none of the outliers would be considered statistically signficantly different.

Table 4. Results falling outside of confidence limits

Food

Category

Food

Type

Sample

Code Food Item

Organism &

Temperature

Spiking/

Seeding

Protocol

Difference

(alternative-

reference)

Dairy & Egg

Products

Pasteurized

Dairy Products 5 Ice Cream

Coliform

30 ± 1°C

Natural

Contamination 0.1139

Dairy & Egg

Products

Raw Dairy

Products 5 Camembert

Coliform

30 ± 1°C

Natural


Dairy & Egg

Products

Desserts &

Egg Products 5 Cream Puffs

Coliform

37 ± 1°C

5 ± 1°C for 48-

72 hours 0.1139

Vegetable

Products

Raw

Vegetables 3 Raddish

Coliform

37 ± 1°C

Natural

Contamination -0.0746

Surface

Samples 4 Air Vent 1

Coliform

37 ± 1°C

Natural


Fish &

Seafood

Products

Raw Products 2 Pollock E. coli

37 ± 1°C

5 ± 1°C for 48-

72 hours -0.3979

Fish &

Seafood

Products

Marinated

Smoked Items 3

Butter & Herb

Cod

E. coli

37 ± 1°C

Natural


Vegetable

Products

Raw

Vegetables 4 Red Bell Pepper

E. coli

37 ± 1°C

Natural


Vegetable

Products

Pre-Cooked

Vegetables 4 Frozen Onions

E. coli

37 ± 1°C

5 ± 1°C for 48-

72 hours -0.3388

RTE & RTRH

Foods

Cold Ready

Meals 1

Chicken Caesar

Wrap

E. coli

37 ± 1°C

Natural


RTE & RTRH

Foods Pasta 3

Pasta with

Broccoli and

Cheddar Sauce

E. coli

37 ± 1°C

Natural


Animal Feed

Finished

Products for

Livestock

2 Chicken Feed E. coli

37 ± 1°C

Natural


Animal Feed

Finished

Products for

Livestock

4 Purina Goat

Grower Feed

E. coli

37 ± 1°C

Natural


Surface

Samples 6 Air Vent

E. coli

37 ± 1°C

Natural


Surface

Samples 14 Cement Table

E. coli

37 ± 1°C

Natural


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Meat Products

Cured and

Smoked

Products (raw)

1 Hot Pork

Sausage

E. coli

42 ± 1°C

Natural


Food

Category

Food

Type

Sample

Code Food Item

Organism &

Temperature

Spiking/

Seeding

Protocol

Difference

(alternative-

reference)

Fish &

Seafood

Products

Marinated

Smoked Items 3

Butter & Herb

Cod

E. coli

42 ± 1°C

Natural


Vegetable

Products

Raw

Vegetables 1 Broccoli

E. coli

42 ± 1°C

Natural


Vegetable

Products

Raw

Vegetables 4 Red Bell Pepper

E. coli

42 ± 1°C

Natural


Vegetable

Products

Pre-Cooked

Vegetables 4 Frozen Onions

E. coli

42 ± 1°C

5°C for 48-72

hours -0.3010

RTE & RTRH

Foods

Cold Ready

Meals 1

Chicken Caesar

Wrap

E. coli

42 ± 1°C

Natural


Animal Feed

Finished

Products for

Domestic

3 Iams E. coli

42 ± 1°C

Natural


Animal Feed

Finished

Products for

Livestock

1 Cow Mana E. coli

42 ± 1°C

Natural


Animal Feed

Finished

Products for

Livestock

4 Purina Goat

Grower Feed

E. coli

42 ± 1°C

Natural


Surface

Sponge 6 Air Vent 3

E. coli

42 ± 1°C

5 ± 1°C for 48-

72 hours -0.3310

Surface

Sponge 7 Pans 1-5

E. coli

42 ± 1°C

5 ± 1°C for 48-

72 hours 0.3010

Surface

Sponge 9 Holding Tank

E. coli

42 ± 1°C

5 ± 1°C for 48-

72 hours 0.3332

Surface

Sponge 14 Cement Table

E. coli

42 ± 1°C

5 ± 1°C for 48-

72 hours -0.3010

3.2.2 Conclusion of the Truness Study

Based on the data obtained in the evaluation, the relative trueness study of the alternative method is considered equivalent to the reference method.

3.3 Accuracy Profile Study

The accuracy profile study is a comparative study between the results obtained by the reference method and the results of the alternative method. The study was conducted using artificially contaminated samples. Two different food types per category were tested.

Number of Samples

For each category being examined, 2 food types were tested at three levels of contamination (low, medium and high). These levels covered the entire range of

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contamination of the selected food type. Artificial contamination was performed using the seeding protocols specified in Table 5 below. For each contamination level, a bulk lot was co-inoculated with both E. coli and coliform and split into 2 samples. From each sample, 5 replicate test portions were evaluated.

Table 5: Summary of Food Categories and Food Items

Food

Category Food Type Food Items

Replicates/

Test Portion

Size1

Inoculating

Organism

(Seeding

protocol)

Target Contamination

Levels

Meat

Products

(Combined

category

raw/RTC

meats and

poultry)

Raw Fresh

Meats2/

Breaded Meats

Minced Meat

&

Raw Frozen

Chicken Wings

15 rep per food item/ 10 g

Klebsiella pneumoniae

& E. coli Liquid

10-100 CFU/g

100-1,000 CFU/g

10,000-100,000 CFU/g

Dairy and

Egg

Products

Raw dairy

products2/Egg

Products (heat

processed) with

additives

Raw milk/whole

liquid egg

15 rep per food item/

10 mL

Hafnia alvei &

E. coli Liquid

10-100 CFU/mL

100-1,000 CFU/mL

10,000-100,000 CFU/mL


Raw

products2/smoked

products

Tuna Sushi/

smoked salmon


E. coli Liquid/stressed

10-100 CFU/g

100-1,000 CFU/g

10,000-100,000 CFU/g

Vegetable Products

Raw vegetable

products/Heat

processed

vegetable juices2

Bunched Spinach/

pasteurized carrot

juice


Klebsiella oxytoca

& E. coli

Liquid/stressed

10-100 CFU/g

100-1,000 CFU/g

10,000-100,000 CFU/g

Ready to Eat

and Ready

to Reheat

Cold Ready

Meals/Mayonnaise

based deli salads

with raw

ingredients2

Ready-made

Sandwiches/

raw vegetable

salad with

dressings.


Citrobacter freundii

& E. coli Liquid

1-10 CFU/g

100-1,000 CFU/g

10,000-100,000 CFU/g

Animal Feed

Finished Products

for Livestock/Plant

Origin Ingredients2

Chicken

Feed/Soybean

Meal


Enterobacter cloacae

& E. coli

lyophilized

10-100 CFU/g

100-1,000 CFU/g

10,000-100,000 CFU/g

Samples

from the

Industrial

Production

Environment

Surface

Samples2,3

Stainless

Environmental

sponges/Sealed

Concrete

Environmental

sponges

15 rep per food item/

Sampling Device

Enterobacter gergoviae

& E. coli Liquid

10-100 CFU/ Sampling Device

100-1,000 CFU/ Sampling Device

10,000-100,000 CFU/ Sampling Device

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1Total number of samples = minimum of 210 (includes paired samples replicates for 3M™ Petrifilm™ and ISO 4832/16649-2) 2 Food type used for frozen plate enumeration evaluation 3Only environmental surface samples will be claimed for this category.

Each matrix was artificially contaminated with a different coliform and E. coli strain. Each inoculum was prepared by transferring a single colony from trypticase soy agar with 5% sheep blood (SBA) into BPW (ISO) and incubating the culture at 35 ± 2oC for 24 ± 2 hours. Following incubation, the culture was diluted to a target level using BPW (ISO) as the diluent. For each inoculated food matrix, bulk portions were spiked and blended in large sterile stainless steel containers. Sterile spatulas were used to mix the bulk portions to ensure the inoculum was evenly distributed throughout the matrix. The minced meat, raw milk, whole liquid eggs, tuna sushi, smoked salmon, bunched spinach, pasteurized carrot juice, ready-made sandwiches, and raw vegetable salad with dressings were held for 48-72 hours at refrigerated temperature (2-8⁰C) prior to analysis to allow time for the organism to equilibrate within the sample. Raw frozen chicken wings were held for two weeks at -20 °C. For chicken feed and soybean meal, a lyophilized inoculum was used to inoculate a bulk lot of each matrix and was then homogenized and held at ambient temperature (20-25⁰C) for two weeks. For surface samples, the inoculatd surface was held at ambient temperature (20-25⁰C) for 16 to 18 hours prior to sampling. Test portions were statistically evaluated according to ISO 16140-2:2016 and MicroVal. The accuracy profile (AP) was used to check the requirement that the alternative method produces a result for a sample that differs from the value produced by the reference method by less than a certain acceptability criterion. Prior to inoculation of the whole liquid eggs and pasteurized carrot juice the broth culture inoculum was heat stressed in a waterbath for 10 ± 1 minute at 50 ± 1°C. The degree of injury was estimated by plating an aliquot of diluted culture onto Violet Red Bile (VRB) agar and NA. The agars were incubated at 35 ± 1°C for 24 ± 2 hours and the colonies enumerated. The degree of injury was estimated as

Where nselect = number of colonies on selective agar and nnonselect = number of colonies on non-selective agar. For one food type of each category, two sets of plates were prepared from the samples. After incubation, one set of plates was placed at -15oC for one week and one set was enumerated immediately. After the frozen storage period, the plates were removed and enumerated. Additionally, from the set of plates stored at -15oC, up to 5 atypical colonies were picked and struck to non-selective agar to demonstrate that further identification from the plates is possible. 3.3.1 Calculation and Data Interpretation

For the Accuracy Profile calculations, the Excel® spreadsheet provided on the ISO website (http://standards.iso.org/iso/16140) was used.

100)1( xn

n

nonselect

select−

http://standards.iso.org/iso/16140

31/92

The accuracy profile was used to check the requirement that the alternative method produces a result for a sample that differs from the value produced by the reference method by less than a certain acceptability criterion. According to the study design, three different levels of contamination were examined. For each sample of every category, the statistical values were calculated in accordance of ISO 16140-2:2016, section 6.1.3.3.

The raw data is provided in Annex H. The statistical analysis and the accuracy profiles are displayed below in Figures 3A-3AL.

3.3.2 Conclusion of the Accuracy Profile Study

The data generated for each category meet the acceptance criteria outlined in ISO 16140-2:2016, section 6.1.3.3. Based on the statistical analysis, the alternative method is considered equivalent to the reference method when immediately enumerated after incubation and can be stored at -15°C for one week. For two food items, raw milk and environmental surface sponges, 1 sample replicate was identified as an outlier causing the AP to fall outside the acceptable limits but within the standard deviation. Since the criteria for additional analysis was not met (sample within standard deviation) no additional statistical analysis was performed. Each of the two outliers just slightly falls outside the CI interval. When removing this single replicate, and replacing it with the means of the other four data points in the concentration level, the method’s performance falls within the acceptable CI’s.

32/92

Figure 3A: Accuracy Statistical Analysis of Raw Fresh Meats – 37 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate

Sample NameReference

central valueBias Lower β-ETI Upper β-ETI

β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.78 0.000 -0.160 0.160 YES YES

1B 1.78 0.000 -0.160 0.160 YES YES

2A 2.78 -0.115 -0.276 0.045 YES YES

2B 2.76 -0.048 -0.209 0.112 YES YES

3A 4.81 0.057 -0.103 0.217 YES YES

3B 4.83 -0.033 -0.193 0.127 YES YES

Reference

method

Alternative

method

SD Repeatability 0.114 0.111 +/- 0.500

(Food) Category Meat Products

(Food) Type Raw Fresh Meats - Coliform 37

SD repeatability of reference

method <= 0.125

YES

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Raw Fresh Meats - Coliform 37 ± 1°C

Bias

β-ETI

AL = +/- 0.5


Central valueBias Lower β-ETI Upper β-ETI

β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.70 0.000 -0.189 0.189 YES YES

1B 1.60 0.000 -0.189 0.189 YES YES

2A 2.41 -0.073 -0.262 0.117 YES YES

2B 2.46 0.000 -0.189 0.189 YES YES

3A 4.58 0.011 -0.178 0.200 YES YES

3B 4.64 -0.138 -0.327 0.051 YES YES

Reference

method

Alternative

method


(Food) Category

(Food) Type

Meat Products

Raw Fresh Meats - E. coli 37 ±


method <= 0.125

YES

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Raw Fresh Meats - E. coli 37 ± 1°C

Bias

β-ETI

AL = +/- 0.5

33/92

Figure 3B: Accuracy Statistical Analysis of Raw Fresh Meats – 42 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.70 0.000 -0.188 0.188 YES YES

1B 1.60 0.000 -0.188 0.188 YES YES

2A 2.41 -0.035 -0.222 0.153 YES YES

2B 2.46 0.015 -0.173 0.202 YES YES

3A 4.58 0.033 -0.155 0.221 YES YES

3B 4.64 -0.087 -0.275 0.101 YES YES

Reference

method

Alternative

method


(Food) Category

(Food) Type

Meat Products



method <= 0.125

YES

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Raw Fresh Meats - E. coli 42 ± 1°C

Bias

β-ETI

AL = +/- 0.5

34/92

Figure 3C: Accuracy Statistical Analysis of Raw Fresh Meats of Frozen Plates – 37 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.78 0.000 -0.147 0.147 YES YES

1B 1.78 0.000 -0.147 0.147 YES YES

2A 2.78 -0.097 -0.244 0.050 YES YES

2B 2.76 -0.008 -0.155 0.140 YES YES

3A 4.81 0.063 -0.084 0.210 YES YES

3B 4.83 -0.033 -0.180 0.114 YES YES

Reference

method

Alternative

method



(Food) Type Raw Fresh Meats - Coliform 37


method <= 0.125

YES

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Raw Fresh Meats - Coliform 37 ± 1°C (Frozen Plates)

Bias

β-ETI

AL = +/- 0.5

35/92

Figure 3D: Accuracy Statistical Analysis of Raw Fresh Meats of Frozen Plates – 42 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.70 -0.097 -0.279 0.085 YES YES

1B 1.60 0.097 -0.085 0.279 YES YES

2A 2.41 0.000 -0.182 0.182 YES YES

2B 2.46 0.015 -0.167 0.197 YES YES

3A 4.58 0.054 -0.128 0.236 YES YES

3B 4.64 -0.075 -0.257 0.107 YES YES

Reference

method

Alternative

method



(Food) Type Raw Fresh Meats - E. coli 42 ±


method <= 0.125

YES

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Raw Fresh Meats - E. coli 42 ± 1°C (Frozen Plates)

Bias

β-ETI

AL = +/- 0.5



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.70 -0.097 -0.277 0.084 YES YES

1B 1.60 0.097 -0.084 0.277 YES YES

2A 2.41 0.000 -0.180 0.180 YES YES

2B 2.46 0.015 -0.166 0.195 YES YES

3A 4.58 0.054 -0.127 0.234 YES YES

3B 4.64 -0.075 -0.256 0.105 YES YES

Reference

method

Alternative

method


(Food) Category

(Food) Type

Meat Products



method <= 0.125

YES

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Raw Fresh Meats - E. coli 37 ± 1°C (Frozen Plates)

Bias

β-ETI

AL = +/- 0.5

36/92

Figure 3E: Accuracy Statistical Analysis of Breaded Meats – 37 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.90 -0.058 -0.269 0.153 YES YES

1B 1.85 0.058 -0.153 0.269 YES YES

2A 2.90 -0.104 -0.315 0.107 YES YES

2B 2.92 0.035 -0.176 0.246 YES YES

3A 4.83 0.081 -0.129 0.292 YES YES

3B 4.87 0.012 -0.199 0.222 YES YES

Reference

method

Alternative

method


Meat Products(Food) Category


method <= 0.125

YES

(Food) Type Breaded Meats - Coliform 37 ±

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Breaded Meats - Coliform 37 ± 1°C

Bias

β-ETI

AL = +/- 0.5



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.70 -0.097 -0.264 0.071 YES YES

1B 1.70 -0.097 -0.264 0.071 YES YES

2A 2.59 -0.060 -0.227 0.108 YES YES

2B 2.61 -0.022 -0.189 0.146 YES YES

3A 4.70 -0.131 -0.298 0.037 YES YES

3B 4.66 0.053 -0.114 0.221 YES YES

Reference

method

Alternative

method


(Food) Type

Meat Products

Breaded Meats - E. coli 37 ±

(Food) Category

YES


method <= 0.125Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Breaded Meats - E. coli 37 ± 1°C

Bias

β-ETI

AL = +/- 0.5

37/92

Figure 3F: Accuracy Statistical Analysis of Breaded Meats – 42 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.70 -0.097 -0.276 0.082 YES YES

1B 1.70 0.000 -0.179 0.179 YES YES

2A 2.59 -0.035 -0.214 0.144 YES YES

2B 2.61 0.010 -0.169 0.189 YES YES

3A 4.70 -0.108 -0.287 0.071 YES YES

3B 4.66 0.062 -0.117 0.241 YES YES

Reference

method

Alternative

method


(Food) Type

Meat Products

Breaded Meats - E. coli 42 ±

(Food) Category

YES



-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

sReference Median

Breaded Meats - E. coli 42 ± 1°C

Bias

β-ETI

AL = +/- 0.5

38/92

Figure 3G: Accuracy Statistical Analysis of Raw Dairy Products – 30 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate

é



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 2.80 -0.021 -0.177 0.135 YES YES

1B 2.90 -0.040 -0.196 0.116 YES YES

2A 3.66 0.062 -0.094 0.217 YES YES

2B 3.76 -0.056 -0.212 0.100 YES YES

3A 4.41 0.000 -0.156 0.156 YES YES

3B 4.45 -0.032 -0.188 0.124 YES YES

Reference

method

Alternative

method

SD Repeatability 0.095 0.108 +/- 0.500YES



Dairy and Egg Products

Raw Dairy Products - Coliform

(Food) Category

(Food) Type

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Raw Dairy Products - Coliform 30 ± 1°C

Bias

β-ETI

AL = +/- 0.5



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 2.66 0.027 -0.205 0.260 YES YES

1B 2.70 0.017 -0.215 0.249 YES YES

2A 3.56 0.000 -0.232 0.232 YES YES

2B 3.53 0.000 -0.232 0.232 YES YES

3A 4.11 -0.269 -0.501 -0.036 NO NO

3B 4.04 0.163 -0.070 0.395 YES YES

Reference

method

Alternative

method


(Food) Category Dairy and Egg Products

(Food) Type Raw Dairy Products - E. coli 30


method <= 0.125

YES

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Raw Dairy Products - E. coli 30 ± 1°C

Bias

β-ETI

AL = +/- 0.5

39/92

Figure 3H: Accuracy Statistical Analysis of Raw Dairy Products of Frozen Plates – 30 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 2.80 0.000 -0.140 0.140 YES YES

1B 2.90 0.005 -0.135 0.145 YES YES

2A 3.66 0.009 -0.131 0.149 YES YES

2B 3.76 -0.015 -0.155 0.125 YES YES

3A 4.41 0.000 -0.140 0.140 YES YES

3B 4.45 0.015 -0.125 0.155 YES YES

Reference

method

Alternative

method


Dairy and Egg Products(Food) Category


method <= 0.125

YES

(Food) Type Raw Dairy Products - Coliform

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Raw Dairy Products - Coliform 30 ± 1°C (Frozen Plates)

Bias

β-ETI

AL = +/- 0.5



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 2.66 -0.040 -0.234 0.155 YES YES

1B 2.70 -0.056 -0.250 0.139 YES YES

2A 3.56 -0.038 -0.233 0.157 YES YES

2B 3.53 0.025 -0.170 0.220 YES YES

3A 4.11 -0.269 -0.464 -0.074 YES YES

3B 4.04 -0.228 -0.423 -0.034 YES YES

Reference

method

Alternative

method


(Food) Type


Raw Dairy Products - E. coli 30

(Food) Category

YES



-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00

Bia

s

Reference Median

Raw Dairy Products - E. coli 30 ± 1°C (Frozen Plates)

Bias

β-ETI

AL = +/- 0.5

40/92

Figure 3I: Accuracy Statistical Analysis of Egg Products – 37 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate

é



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.90 -0.125 -0.294 0.044 YES YES

1B 1.78 0.125 -0.044 0.294 YES YES

2A 2.77 0.014 -0.154 0.183 YES YES

2B 2.83 -0.027 -0.196 0.142 YES YES

3A 4.89 0.022 -0.147 0.191 YES YES

3B 4.89 0.022 -0.147 0.191 YES YES

Reference

method

Alternative

method

(Food) Type


Final AL

Egg Products - Coliform 37 ±


method <= 0.125

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Egg Products - Coliform 37 ± 1°C

Bias

β-ETI

AL = +/- 0.5



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.70 -0.097 -0.218 0.024 YES YES

1B 1.60 0.097 -0.024 0.218 YES YES

2A 2.61 0.120 -0.002 0.241 YES YES

2B 2.66 -0.040 -0.161 0.082 YES YES

3A 4.69 -0.077 -0.199 0.044 YES YES

3B 4.73 -0.201 -0.322 -0.080 YES YES

Reference

method

Alternative

method



method <= 0.125

YES


Final AL

(Food) Type Egg Products - E. coli 37 ± 1°C

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Egg Products - E. coli 37 ± 1°C

Bias

β-ETI

AL = +/- 0.5

41/92

Figure 3J: Accuracy Statistical Analysis of Egg Products – 42 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.70 0.000 -0.118 0.118 YES YES

1B 1.60 0.097 -0.021 0.215 YES YES

2A 2.61 0.111 -0.007 0.230 YES YES

2B 2.66 0.000 -0.118 0.118 YES YES

3A 4.69 -0.057 -0.175 0.062 YES YES

3B 4.73 0.016 -0.103 0.134 YES YES

Reference

method

Alternative

method



method <= 0.125

YES


Final AL

(Food) Type Egg Products - E. coli 42 ± 1°C

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

sReference Median

Egg Products - E. coli 42 ± 1°C

Bias

β-ETI

AL = +/- 0.5

42/92

Figure 3K: Accuracy Statistical Analysis of Raw Fish and Seafood Products – 37 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.85 0.155 0.037 0.273 YES YES

1B 1.95 0.000 -0.118 0.118 YES YES

2A 2.90 -0.022 -0.141 0.096 YES YES

2B 2.91 0.036 -0.082 0.154 YES YES

3A 4.85 -0.006 -0.125 0.112 YES YES

3B 4.90 -0.034 -0.152 0.085 YES YES

Reference

method

Alternative

method


(Food) Category Fish And Seafood Products

(Food) Type Raw Products - Coliform 37 ±


method <= 0.125

YES

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Raw Products - Coliform 37 ± 1°C

Bias

β-ETI

AL = +/- 0.5



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.70 -0.097 -0.247 0.053 YES YES

1B 1.70 0.000 -0.150 0.150 YES YES

2A 2.56 0.000 -0.150 0.150 YES YES

2B 2.64 -0.064 -0.214 0.086 YES YES

3A 4.58 -0.048 -0.198 0.102 YES YES

3B 4.58 -0.012 -0.162 0.139 YES YES

Reference

method

Alternative

method


(Food) Category

(Food) Type

Fish And Seafood Products

Raw Products - E. coli 37 ± 1°C


method <= 0.125

YES

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median


Bias

β-ETI

AL = +/- 0.5

43/92

Figure 3L: Accuracy Statistical Analysis of Raw Fish and Seafood Products – 42 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.70 0.000 -0.326 0.326 YES YES

1B 1.70 0.000 -0.326 0.326 YES YES

2A 2.56 0.023 -0.303 0.350 YES YES

2B 2.64 -0.041 -0.368 0.285 YES YES

3A 4.58 -0.023 -0.350 0.303 YES YES

3B 4.58 0.011 -0.315 0.338 YES YES

Reference

method

Alternative

method



method <= 0.125

YES

Final AL

(Food) Category

(Food) Type



-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median


Bias

β-ETI

AL = +/- 0.5

44/92

Figure 3M: Accuracy Statistical Analysis of Raw Fish and Seafood Products of Frozen Plates– 37 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate

é



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.85 0.109 0.005 0.213 YES YES

1B 1.95 -0.051 -0.155 0.053 YES YES

2A 2.90 -0.005 -0.109 0.098 YES YES

2B 2.91 0.011 -0.093 0.115 YES YES

3A 4.85 -0.013 -0.117 0.091 YES YES

3B 4.90 0.011 -0.093 0.115 YES YES

Reference

method

Alternative

method





Raw Products - Coliform 37 ±

(Food) Category

(Food) Type

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Raw Products - Coliform 37 ± 1°C (Frozen Plates)

Bias

β-ETI

AL = +/- 0.5



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.70 0.000 -0.110 0.110 YES YES

1B 1.70 0.000 -0.110 0.110 YES YES

2A 2.56 -0.038 -0.148 0.072 YES YES

2B 2.64 -0.075 -0.185 0.034 YES YES

3A 4.58 -0.048 -0.158 0.061 YES YES

3B 4.58 -0.023 -0.133 0.086 YES YES

Reference

method

Alternative

method


(Food) Category Fish And Seafood Products

(Food) Type Raw Products - E. coli 37 ± 1°C


method <= 0.125

YES

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00

Bia

s

Reference Median

Raw Products - E. coli 37 ± 1°C (Frozen Plates)

Bias

β-ETI

AL = +/- 0.5

45/92

Figure 3N: Accuracy Statistical Analysis of Raw Fish and Seafood Products of Frozen Plates– 42 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.70 0.079 -0.036 0.195 YES YES

1B 1.70 0.079 -0.036 0.195 YES YES

2A 2.56 0.067 -0.049 0.182 YES YES

2B 2.64 -0.052 -0.168 0.063 YES YES

3A 4.58 -0.012 -0.127 0.104 YES YES

3B 4.58 0.011 -0.104 0.127 YES YES

Reference

method

Alternative

method


(Food) Category

(Food) Type




method <= 0.125

YES

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Raw Products - E. coli 42 ± 1°C (Frozen Plates)

Bias

β-ETI

AL = +/- 0.5

46/92

Figure 3O: Accuracy Statistical Analysis of Smoked Fish and Seafood Products – 37 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.90 -0.058 -0.189 0.073 YES YES

1B 1.90 0.000 -0.131 0.131 YES YES

2A 2.84 0.085 -0.046 0.217 YES YES

2B 2.89 -0.006 -0.137 0.126 YES YES

3A 4.88 -0.042 -0.173 0.089 YES YES

3B 4.78 0.014 -0.117 0.146 YES YES

Reference

method

Alternative

method


Fish And Seafood Products(Food) Category


method <= 0.125

YES

(Food) Type Smoked Products - Coliform 37

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Smoked Products - Coliform 37 ± 1°C

Bias

β-ETI

AL = +/- 0.5



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.70 -0.097 -0.241 0.047 YES YES

1B 1.70 -0.097 -0.241 0.047 YES YES

2A 2.56 -0.038 -0.182 0.107 YES YES

2B 2.48 0.000 -0.144 0.144 YES YES

3A 4.56 -0.094 -0.238 0.050 YES YES

3B 4.49 0.000 -0.144 0.144 YES YES

Reference

method

Alternative

method


(Food) Type


Smoked Products - E. coli 37 ±

(Food) Category

YES



-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Smoked Products - E. coli 37 ± 1°C

Bias

β-ETI

AL = +/- 0.5

47/92

Figure 3P: Accuracy Statistical Analysis of Smoked Fish and Seafood Products – 42 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.70 0.000 -0.144 0.144 YES YES

1B 1.70 -0.097 -0.241 0.047 YES YES

2A 2.56 0.000 -0.144 0.144 YES YES

2B 2.48 0.014 -0.130 0.159 YES YES

3A 4.56 -0.065 -0.209 0.079 YES YES

3B 4.49 0.027 -0.117 0.172 YES YES

Reference

method

Alternative

method




(Food) Category

YES

(Food) Type


Smoked Products - E. coli 42 ±

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Smoked Products - E. coli 42 ± 1°C

Bias

β-ETI

AL = +/- 0.5

48/92

3Q: Accuracy Statistical Analysis of Raw Vegetable Products – 37 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate

é



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 2.80 -0.007 -0.104 0.090 YES YES

1B 2.76 -0.008 -0.104 0.089 YES YES

2A 3.59 0.000 -0.097 0.097 YES YES

2B 3.61 -0.045 -0.141 0.052 YES YES

3A 4.45 -0.016 -0.113 0.081 YES YES

3B 4.45 -0.016 -0.113 0.081 YES YES

Reference

method

Alternative

method




Vegetable Products

Raw Vegetable Products -

(Food) Category

(Food) Type

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Raw Vegetable Products - Coliform 37 ± 1°C

Bias

β-ETI

AL = +/- 0.5



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 2.43 -0.070 -0.254 0.115 YES YES

1B 2.46 -0.047 -0.232 0.137 YES YES

2A 3.36 -0.061 -0.245 0.124 YES YES

2B 3.38 -0.176 -0.361 0.009 YES YES

3A 4.15 -0.105 -0.290 0.080 YES YES

3B 4.08 0.000 -0.185 0.185 YES YES

Reference

method

Alternative

method


(Food) Category Vegetable Products

(Food) Type Raw Vegetable Products - E.


method <= 0.125

YES

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Raw Vegetable Products - E. coli 37 ± 1°C

Bias

β-ETI

AL = +/- 0.5

49/92

3R: Accuracy Statistical Analysis of Raw Vegetable Products – 42 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 2.43 -0.016 -0.162 0.129 YES YES

1B 2.46 -0.031 -0.177 0.115 YES YES

2A 3.36 -0.019 -0.165 0.127 YES YES

2B 3.38 -0.125 -0.271 0.021 YES YES

3A 4.15 -0.067 -0.213 0.079 YES YES

3B 4.08 0.035 -0.111 0.181 YES YES

Reference

method

Alternative

method



(Food) Type Raw Vegetable Products - E.


method <= 0.125

YES

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

sReference Median

Raw Vegetable Products - E. coli 42 ± 1°C

Bias

β-ETI

AL = +/- 0.5

50/92

Figure 3S: Accuracy Statistical Analysis of Heat Processed Vegetable Juices – 37 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate

é



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.90 0.051 -0.067 0.170 YES YES

1B 1.95 -0.109 -0.228 0.009 YES YES

2A 2.91 0.005 -0.113 0.124 YES YES

2B 2.92 -0.038 -0.156 0.081 YES YES

3A 4.91 0.016 -0.103 0.134 YES YES

3B 4.89 -0.006 -0.124 0.113 YES YES

Reference

method

Alternative

method


(Food) Type


Final AL

Heat Processed Vegetable


method <= 0.125

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Heat Processed Vegetable Juices - Coliform 37 ± 1°C

Bias

β-ETI

AL = +/- 0.5



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.78 -0.176 -0.351 -0.001 YES YES

1B 1.70 -0.097 -0.272 0.078 YES YES

2A 2.72 0.000 -0.175 0.175 YES YES

2B 2.75 -0.135 -0.310 0.039 YES YES

3A 4.56 0.012 -0.163 0.187 YES YES

3B 4.56 0.023 -0.151 0.198 YES YES

Reference

method

Alternative

method



method <= 0.125

YES


Final AL

(Food) Type Heat Processed Vegetable

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Heat Processed Vegetable Juices - E. coli 37 ± 1°C

Bias

β-ETI

AL = +/- 0.5

51/92

Figure 3T: Accuracy Statistical Analysis of Heat Processed Vegetable Juices – 42 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.78 -0.079 -0.252 0.094 YES YES

1B 1.70 0.079 -0.094 0.252 YES YES

2A 2.72 0.032 -0.141 0.205 YES YES

2B 2.75 -0.115 -0.288 0.059 YES YES

3A 4.56 0.023 -0.150 0.197 YES YES

3B 4.56 0.046 -0.127 0.219 YES YES

Reference

method

Alternative

method



Final AL



method <= 0.125

YES

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Heat Processed Vegetable Juices - E. coli 42 ± 1°C

Bias

β-ETI

AL = +/- 0.5

52/92

Figure 3U: Accuracy Statistical Analysis of Heat Processed Vegetable Juices of Frozen Plates– 37 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate

é



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.90 0.000 -0.104 0.104 YES YES

1B 1.95 -0.051 -0.155 0.053 YES YES

2A 2.91 -0.005 -0.109 0.099 YES YES

2B 2.92 0.010 -0.094 0.114 YES YES

3A 4.91 0.016 -0.088 0.120 YES YES

3B 4.89 -0.023 -0.127 0.081 YES YES

Reference

method

Alternative

method


(Food) Type


Final AL

Heat Processed Vegetable


method <= 0.125

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Heat Processed Vegetable Juices - Coliform 37 ± 1°C (Frozen Plates)

Bias

β-ETI

AL = +/- 0.5



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.78 -0.079 -0.209 0.051 YES YES

1B 1.70 -0.097 -0.227 0.033 YES YES

2A 2.72 -0.017 -0.147 0.113 YES YES

2B 2.75 -0.049 -0.179 0.081 YES YES

3A 4.56 -0.025 -0.155 0.105 YES YES

3B 4.56 0.012 -0.118 0.142 YES YES

Reference

method

Alternative

method



method <= 0.125

YES


Final AL


-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Heat Processed Vegetable Juices - E. coli 37 ± 1°C (Frozen Plates)

Bias

β-ETI

AL = +/- 0.5

53/92

Figure 3V: Accuracy Statistical Analysis of Heat Processed Vegetable Juices of Frozen Plates– 42 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.78 0.000 -0.115 0.115 YES YES

1B 1.70 0.079 -0.036 0.195 YES YES

2A 2.72 0.016 -0.099 0.132 YES YES

2B 2.75 -0.024 -0.139 0.092 YES YES

3A 4.56 0.023 -0.092 0.139 YES YES

3B 4.56 0.035 -0.081 0.150 YES YES

Reference

method

Alternative

method


Vegetable Products(Food) Category


method <= 0.125

YES


Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Heat Processed Vegetable Juices - E. coli 42 ± 1°C (Frozen Plates)

Bias

β-ETI

AL = +/- 0.5

54/92

Figure 3W: Accuracy Statistical Analysis of Cold Ready Meals – 37 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.90 0.000 -0.101 0.101 YES YES

1B 1.90 0.000 -0.101 0.101 YES YES

2A 2.86 0.017 -0.084 0.119 YES YES

2B 2.90 -0.006 -0.107 0.096 YES YES

3A 4.87 -0.030 -0.131 0.071 YES YES

3B 4.89 -0.041 -0.142 0.060 YES YES

Reference

method

Alternative

method



method <= 0.125

YES

Final AL

(Food) Category RTE/RTRH

(Food) Type Cold Ready Meals - Coliform 37

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Cold Ready Meals - Coliform 37 ± 1°C

Bias

β-ETI

AL = +/- 0.5



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.78 -0.176 -0.320 -0.032 YES YES

1B 1.70 -0.097 -0.241 0.047 YES YES

2A 2.48 -0.015 -0.159 0.130 YES YES

2B 2.43 0.087 -0.057 0.232 YES YES

3A 4.56 -0.051 -0.196 0.093 YES YES

3B 4.60 -0.111 -0.255 0.034 YES YES

Reference

method

Alternative

method



method <= 0.125

YES

Final AL

(Food) Type Cold Ready Meals - E. coli 37 ±

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Cold Ready Meals - E. coli 37 ± 1°C

Bias

β-ETI

AL = +/- 0.5

55/92

Figure 3X: Accuracy Statistical Analysis of Cold Ready Meals – 42 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.78 -0.079 -0.232 0.074 YES YES

1B 1.70 0.000 -0.153 0.153 YES YES

2A 2.48 0.028 -0.125 0.181 YES YES

2B 2.43 0.125 -0.028 0.278 YES YES

3A 4.56 0.000 -0.153 0.153 YES YES

3B 4.60 -0.071 -0.224 0.082 YES YES

Reference

method

Alternative

method


(Food) Category

(Food) Type

RTE/RTRH

Cold Ready Meals - E. coli 42 ±


method <= 0.125

YES

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

sReference Median

Cold Ready Meals - E. coli 42 ± 1°C

Bias

β-ETI

AL = +/- 0.5

56/92

Figure 3Y: Accuracy Statistical Analysis of Mayonnaise based Deli Salad with Raw Ingredients – 37 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.90 -0.058 -0.175 0.059 YES YES

1B 1.90 0.000 -0.117 0.117 YES YES

2A 2.93 -0.015 -0.132 0.102 YES YES

2B 2.95 -0.063 -0.180 0.054 YES YES

3A 4.87 -0.037 -0.154 0.080 YES YES

3B 4.86 0.000 -0.117 0.117 YES YES

Reference

method

Alternative

method



method <= 0.125

YES

(Food) Type Mayonnaise based deli salads

Final AL

RTE/RTRH(Food) Category

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Mayonnaise based deli salads with raw ingredients -Coliform 37 ± 1°C

Bias

β-ETI

AL = +/- 0.5



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.60 0.000 -0.159 0.159 YES YES

1B 1.60 0.000 -0.159 0.159 YES YES

2A 2.66 -0.019 -0.178 0.139 YES YES

2B 2.68 -0.038 -0.197 0.121 YES YES

3A 4.48 -0.015 -0.174 0.144 YES YES

3B 4.51 -0.058 -0.217 0.101 YES YES

Reference

method

Alternative

method




(Food) Category

YES

(Food) Type

RTE/RTRH

Mayonnaise based deli salads

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Mayonnaise based deli salads with raw ingredients - E. coli 37 ± 1°C

Bias

β-ETI

AL = +/- 0.5

57/92

Figure 3Z: Accuracy Statistical Analysis of Mayonnaise based Deli Salad with Raw Ingredients – 42 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.60 0.000 -0.165 0.165 YES YES

1B 1.60 0.000 -0.165 0.165 YES YES

2A 2.66 0.018 -0.146 0.183 YES YES

2B 2.68 -0.018 -0.183 0.146 YES YES

3A 4.48 0.000 -0.165 0.165 YES YES

3B 4.51 -0.074 -0.238 0.091 YES YES

Reference

method

Alternative

method


(Food) Type

RTE/RTRH


(Food) Category

YES



-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Mayonnaise based deli salads with raw ingredients - E. coli 42 ± 1°C

Bias

β-ETI

AL = +/- 0.5

58/92

Figure 3AA: Accuracy Statistical Analysis of Mayonnaise based Deli Salad with Raw Ingredients of Frozen Plates– 37 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.90 0.000 -0.098 0.098 YES YES

1B 1.90 -0.058 -0.156 0.040 YES YES

2A 2.93 -0.010 -0.108 0.088 YES YES

2B 2.95 -0.010 -0.108 0.088 YES YES

3A 4.87 -0.018 -0.116 0.080 YES YES

3B 4.86 0.006 -0.092 0.104 YES YES

Reference

method

Alternative

method


(Food) Category RTE/RTRH

(Food) Type Mayonnaise based deli salads


method <= 0.125

YES

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Mayonnaise based deli salads with raw ingredients -Coliform 37 ± 1°C (Frozen Plates)

Bias

β-ETI

AL = +/- 0.5



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.60 0.000 -0.169 0.169 YES YES

1B 1.60 0.000 -0.169 0.169 YES YES

2A 2.66 -0.019 -0.188 0.150 YES YES

2B 2.68 -0.048 -0.217 0.121 YES YES

3A 4.48 -0.046 -0.215 0.123 YES YES

3B 4.51 -0.090 -0.259 0.079 YES YES

Reference

method

Alternative

method


(Food) Category

(Food) Type

RTE/RTRH



method <= 0.125

YES

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Mayonnaise based deli salads with raw ingredients - E. coli 37 ± 1°C (Frozen Plates)

Bias

β-ETI

AL = +/- 0.5

59/92

Figure 3AB: Accuracy Statistical Analysis of Mayonnaise based Deli Salad with Raw Ingredients of Frozen Plates– 42 ± 1°C 3M

Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.60 0.097 -0.058 0.251 YES YES

1B 1.60 0.176 0.022 0.331 YES YES

2A 2.66 0.027 -0.127 0.182 YES YES

2B 2.68 0.009 -0.146 0.163 YES YES

3A 4.48 0.000 -0.154 0.154 YES YES

3B 4.51 -0.028 -0.182 0.126 YES YES

Reference

method

Alternative

method


(Food) Type

RTE/RTRH


(Food) Category

YES



-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Mayonnaise based deli salads with raw ingredients - E. coli 42 ± 1°C (Frozen Plates)

Bias

β-ETI

AL = +/- 0.5

60/92

Figure 3AC: Accuracy Statistical Analysis of Finished Products for Livestock – 37 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate

é



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.90 0.000 -0.101 0.101 YES YES

1B 1.85 0.058 -0.043 0.159 YES YES

2A 2.91 0.011 -0.090 0.112 YES YES

2B 2.94 -0.053 -0.154 0.048 YES YES

3A 4.91 0.026 -0.075 0.127 YES YES

3B 4.89 0.053 -0.048 0.154 YES YES

Reference

method

Alternative

method


Animal Feed

Finished Products for Livestock -

(Food) Category

(Food) Type



YES

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Finished Products for Livestock - Coliform 37 ± 1°C

Bias

β-ETI

AL = +/- 0.5



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.85 -0.243 -0.353 -0.133 YES YES

1B 1.78 -0.176 -0.286 -0.066 YES YES

2A 2.71 -0.017 -0.127 0.092 YES YES

2B 2.69 0.026 -0.084 0.136 YES YES

3A 4.59 -0.100 -0.209 0.010 YES YES

3B 4.61 -0.094 -0.204 0.015 YES YES

Reference

method

Alternative

method


(Food) Category Animal Feed

(Food) Type Finished Products for Livestock -


method <= 0.125

YES

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Finished Products for Livestock - E. coli 37 ± 1°C

Bias

β-ETI

AL = +/- 0.5

61/92

Figure 3AD: Accuracy Statistical Analysis of Finished Products for Livestock – 42 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.85 -0.243 -0.376 -0.110 YES YES

1B 1.78 -0.176 -0.309 -0.043 YES YES

2A 2.71 0.000 -0.133 0.133 YES YES

2B 2.69 0.042 -0.091 0.175 YES YES

3A 4.59 -0.060 -0.192 0.073 YES YES

3B 4.61 0.000 -0.133 0.133 YES YES

Reference

method

Alternative

method



(Food) Type Finished Products for Livestock -


method <= 0.125

YES

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Finished Products for Livestock - E. coli 42 ± 1°C

Bias

β-ETI

AL = +/- 0.5

62/92

Figure 3AE: Accuracy Statistical Analysis of Plant Origin Ingredients – 37 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate

é



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.85 0.000 -0.127 0.127 YES YES

1B 1.90 0.000 -0.127 0.127 YES YES

2A 2.92 -0.038 -0.165 0.089 YES YES

2B 2.91 0.016 -0.111 0.143 YES YES

3A 4.90 -0.059 -0.186 0.068 YES YES

3B 4.88 0.006 -0.121 0.133 YES YES

Reference

method

Alternative

method



Final AL

Plant Origin Ingredients -


method <= 0.125

YES

(Food) Type

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Plant Origin Ingredients - Coliform 37 ± 1°C

Bias

β-ETI

AL = +/- 0.5



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.60 0.000 -0.114 0.114 YES YES

1B 1.78 -0.176 -0.290 -0.062 YES YES

2A 2.71 -0.064 -0.178 0.050 YES YES

2B 2.69 0.009 -0.105 0.123 YES YES

3A 4.62 -0.092 -0.206 0.022 YES YES

3B 4.58 -0.075 -0.189 0.039 YES YES

Reference

method

Alternative

method



Final AL

(Food) Type Plant Origin Ingredients - E. coli


method <= 0.125

YES

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Plant Origin Ingredients - E. coli 37 ± 1°C

Bias

β-ETI

AL = +/- 0.5

63/92

Figure 3AF: Accuracy Statistical Analysis of Plant Origin Ingredients – 42 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.60 0.000 -0.133 0.133 YES YES

1B 1.78 -0.176 -0.309 -0.043 YES YES

2A 2.71 -0.017 -0.150 0.115 YES YES

2B 2.69 0.026 -0.107 0.159 YES YES

3A 4.62 -0.043 -0.176 0.089 YES YES

3B 4.58 -0.048 -0.181 0.085 YES YES

Reference

method

Alternative

method



method <= 0.125

YES


Final AL

(Food) Type Plant Origin Ingredients - E. coli

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

sReference Median

Plant Origin Ingredients - E. coli 42 ± 1°C

Bias

β-ETI

AL = +/- 0.5

64/92

Figure 3AG: Accuracy Statistical Analysis of Plant Origin Ingredients of Frozen Plates – 37 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.85 0.058 -0.046 0.162 YES YES

1B 1.90 -0.058 -0.162 0.046 YES YES

2A 2.92 -0.011 -0.115 0.093 YES YES

2B 2.91 0.005 -0.099 0.109 YES YES

3A 4.90 -0.011 -0.115 0.093 YES YES

3B 4.88 0.000 -0.104 0.104 YES YES

Reference

method

Alternative

method


Animal Feed(Food) Category


method <= 0.125

YES

(Food) Type Plant Origin Ingredients -

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Plant Origin Ingredients - Coliform 37 ± 1°C (Frozen Plates)

Bias

β-ETI

AL = +/- 0.5



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.60 0.000 -0.152 0.152 YES YES

1B 1.78 -0.079 -0.231 0.072 YES YES

2A 2.71 -0.026 -0.178 0.125 YES YES

2B 2.69 -0.027 -0.179 0.124 YES YES

3A 4.62 -0.105 -0.256 0.047 YES YES

3B 4.58 -0.075 -0.226 0.077 YES YES

Reference

method

Alternative

method


(Food) Type

Animal Feed

Plant Origin Ingredients - E. coli

(Food) Category

YES



-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Plant Origin Ingredients - E. coli 37 ± 1°C (Frozen Plates)

Bias

β-ETI

AL = +/- 0.5

65/92

Figure 3AH: Accuracy Statistical Analysis of Plant Origin Ingredients of Frozen Plates – 42 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate

é



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.60 0.097 -0.053 0.247 YES YES

1B 1.78 0.000 -0.150 0.150 YES YES

2A 2.71 0.017 -0.133 0.167 YES YES

2B 2.69 0.009 -0.141 0.159 YES YES

3A 4.62 -0.032 -0.182 0.118 YES YES

3B 4.58 -0.048 -0.198 0.102 YES YES

Reference

method

Alternative

method




Animal Feed

Plant Origin Ingredients - E. coli

(Food) Category

(Food) Type

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Plant Origin Ingredients - E. coli 42 ± 1°C (Frozen Plates)

Bias

β-ETI

AL = +/- 0.5

66/92

3AI: Accuracy Statistical Analysis of Surface Samples – 37 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.85 0.000 -0.195 0.195 YES YES

1B 2.72 -0.026 -0.221 0.169 YES YES

2A 2.92 -0.027 -0.222 0.168 YES YES

2B 3.62 -0.043 -0.238 0.151 YES YES

3A 4.46 -0.161 -0.356 0.034 YES YES

3B 4.86 -0.072 -0.267 0.123 YES YES

Reference

method

Alternative

method



method <= 0.125

YES

Final AL

(Food) Category

(Food) Type

Samples from the Industrial

Surface Samples - Coliform 37 ±

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Surface Samples - Coliform 37 ± 1°C

Bias

β-ETI

AL = +/- 0.5



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.70 -0.097 -0.332 0.138 YES YES

1B 2.36 -0.248 -0.483 -0.012 YES YES

2A 2.56 -0.065 -0.300 0.170 YES YES

2B 2.88 0.265 0.030 0.501 NO NO

3A 4.00 0.150 -0.085 0.386 YES YES

3B 4.49 -0.111 -0.346 0.124 YES YES

Reference

method

Alternative

method




(Food) Category

YES

(Food) Type


Surface Samples - E. coli 37 ±

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Surface Samples - E. coli 37 ± 1°C

Bias

β-ETI

AL = +/- 0.5

67/92

3AJ: Accuracy Statistical Analysis of Surface Samples – 42 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.70 -0.097 -0.324 0.130 YES YES

1B 2.36 -0.158 -0.384 0.069 YES YES

2A 2.56 -0.025 -0.251 0.202 YES YES

2B 2.91 -0.096 -0.322 0.131 YES YES

3A 3.99 0.155 -0.072 0.382 YES YES

3B 4.49 -0.044 -0.271 0.182 YES YES

Reference

method

Alternative

method


(Food) Category Samples from the Industrial

(Food) Type Surface Samples - E. coli 42 ±


method <= 0.125

YES

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

sReference Median

Surface Samples - E. coli 42 ± 1°C

Bias

β-ETI

AL = +/- 0.5

68/92

Figure 3AK: Accuracy Statistical Analysis of Surface Samples of Frozen Plates – 37 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate

é



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.85 0.000 -0.105 0.105 YES YES

1B 2.72 0.008 -0.097 0.114 YES YES

2A 2.92 -0.061 -0.166 0.044 YES YES

2B 3.62 -0.010 -0.116 0.095 YES YES

3A 4.46 -0.082 -0.188 0.023 YES YES

3B 4.86 -0.018 -0.124 0.087 YES YES

Reference

method

Alternative

method



Surface Samples - Coliform 37 ±

(Food) Category

(Food) Type



YES

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Surface Samples - Coliform 37 ± 1°C (Frozen Plates)

Bias

β-ETI

AL = +/- 0.5



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.70 -0.097 -0.221 0.027 YES YES

1B 2.36 -0.186 -0.310 -0.061 YES YES

2A 2.56 -0.094 -0.218 0.030 YES YES

2B 2.88 -0.103 -0.227 0.021 YES YES

3A 4.00 -0.217 -0.342 -0.093 YES YES

3B 4.49 -0.130 -0.254 -0.005 YES YES

Reference

method

Alternative

method


(Food) Category Samples from the Industrial

(Food) Type Surface Samples - E. coli 37 ±


method <= 0.125

YES

Final AL

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Surface Samples - E. coli 37 ± 1°C (Frozen Plates)

Bias

β-ETI

AL = +/- 0.5

69/92

Figure 3AL: Accuracy Statistical Analysis of Surface Samples of Frozen Plates – 42 ± 1°C 3M Petrifilm Rapid E. coli / Coliform Count Plate



β-ETI

compared to

AL=±0.5

Acceptable

β-ETI

compared to

final AL

Acceptable

1A 1.70 0.000 -0.175 0.175 YES YES

1B 2.36 -0.040 -0.214 0.135 YES YES

2A 2.56 -0.025 -0.200 0.150 YES YES

2B 2.91 0.296 0.121 0.470 YES YES

3A 3.99 -0.146 -0.321 0.029 YES YES

3B 4.49 -0.029 -0.204 0.146 YES YES

Reference

method

Alternative

method




(Food) Category

YES

(Food) Type


Surface Samples - E. coli 42 ±

-0.60

-0.40

-0.20

0.00

0.20

0.40

0.60

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Bia

s

Reference Median

Surface Samples - E. coli 42 ± 1°C (Frozen Plates)

Bias

β-ETI

AL = +/- 0.5

70/92

3.4 Inclusivity and Inclusivity and Exclusivity Study

The inclusivity study involved pure target strains to be detected by the alternative method. The exclusivity study involved pure non-target strains, which can be potentially cross-reactive, but are not expected to be detected by the alternative method. See Table 6-9 for the list of strains. 50 target strains of coliform and E. coli and 30 non-target strains of coliform and E. coli were tested. All isolates were enriched in non-selective broth for 18-24 hours at 37 ± 1ºC. Ten fold serial dilutions were conducted in order to achieve a final concentration of ~50 cells/mL. A 1.0 mL aliquot from this dilution was plated onto three sets of plates. Individual plates were incubated for 18 hours at 30 ± 1ºC, 37 ± 1ºC and 42 ± 1ºC.

Table 6: E. coli Inclusivity Strains

Number Genus Species Source Origin

1 Escherichia coli ATCC1 8739 Feces

2 Escherichia coli NBRC2 15034 Clinical Specimen

3 Escherichia coli ATCC 25922 Not Available

4 Escherichia coli ATCC 11229 feces



7 Escherichia coli ATCC 11775 food, Minnesota

8 Escherichia coli Q Labs 11010-2 Bottled Water




12 Escherichia coli ATCC 35218 canine, Tennessee


14 Escherichia coli Q Labs 41411.1 Bottled water

15 Escherichia coli Q Labs 41411.2 Bottled water

16 Escherichia coli Q Labs16154-1 Beef Carcass Sponge

17 Escherichia coli Q Labs 16154-2 Raw Ground Beef

18 Escherichia coli Q Labs 16154-3 Beef Trim

19 Escherichia coli Q Labs 16154-5 Flavored Seasoning

20 Escherichia coli Q Labs 16154-7 Flour

21 Escherichia coli Q Labs 16154-12 Chicken Carcass Rinse

22 Escherichia coli Q Labs 16154-13 Potable Water

23 Escherichia coli Q Labs 16154-15 Raw Chicken Breast

24 Escherichia coli Q Labs 16160-2 Mixed Frozen Vegetables

25 Escherichia coli Q Labs 16160-3 Frozen Greens

26 Escherichia coli Q Labs 16160-4 Chicken Nuggets


28 Escherichia coli Q Labs16160-8 Chicken Carcass Rinse

29 Escherichia coli Q Labs 16160-9 Fresh Baby Spinach

30 Escherichia coli Q Labs 16160-10 Raw Pork

31 Escherichia coli Q Labs 16160-11 Raw Beef Trimmings

32 Escherichia coli Q Labs 16160-13 Irrigation Rinse Water

33 Escherichia coli Q Labs 16160-16 Sprouts

34 Escherichia coli Q Labs 16160-17 Fresh Spinach

35 Escherichia coli Q Labs 16160-18 Frozen Mixed Vegetables

36 Escherichia coli 3M4 REC1 TBP5

37 Escherichia coli 3M REC5 TBP

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50 Escherichia coli 3M REC19 TBP 1ATCC-American Type Culture Collection, 2NBRC-Nite Biological Resource Center, 3QL- Q Laboratories Culture Collection, 43M – 3M Food Safety, 5. TBP- To be provided by 3M Food Safety

Table 7: E. coli Exclusivity Strains


1 Acinetobacter baumanii ATCC1 19606 Urine

2 Aeromonas viridans QL2 17041-8 Raw Milk Isolate

3 Alcaligenes faecalis ATCC 8750 Not Available

4 Bordetella bronchiseptica ATCC 10580 Lung of dog

5 Brochothrix thermosphacta ATCC 11509 Animal-derived foodstuff

6 Carnobacterium maltaromaticum ATCC 43224 Vacuum Packed Beef

7 Citrobacter freundii QL 100813-2A Deli Meat

8 Citrobacter farmeri ATCC 51613 Clinical Isolate

9 Cronobacter condimenti QL 17031.1 Infant Formula

10 Cronobacter sakazakii QL 11007-9 Rice Flour

11 Enterococcus faecium ATCC 516 Clinical isolate

12 Haemophilus influenzae ATCC 19418 Not Available

13 Hafnia alvei ATCC 51815 Milk

14 Kurthia gibsonii ATCC 43195 RTE meat

15 Klebsiella oxytoca ATCC 43165 Clinical

16 Klebsiella pneumoniae QL 1100-7 Raw meat

17 Kluyvera Intermedia ATCC 33110 Surface Water

18 Microbacterium testaceum ATCC 15829 Paddy

19 Morganella morganii ATCC 25829 Human isolate

20 Pantoea agglomerans ATCC 19552 sewage

21 Proteus mirabilis QL 11007-6 Animal specimen

22 Providencia stuartii QL 11007-5 Clinical isolate

23 Pseudomonas extremorientialis QL 17041-1 Raw Milk Isolate

24 Pseudomonas fluorescens QL 17041-3 Raw Milk Isolate

25 Rahnella aquatilis QL 14181-2A Soy protein

26 Salmonella Typhimurium QL 011414-2 Raw material

27 Serratia marcescens QL 11007-1 Bottled water

28 Streptococcus pyogenes ATCC 19615 Pharynx of child following sore

throat.

29 Sphingomonas paucimobilis ATCC 29837 Hospital Respirator

30 Vibrio vulnificus QL 021111A Seafood Product 1ATCC-American Type Culture Collection, 2QL-Q Laboratories Culture Collection.

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Table 8: Total Coliform Inclusivity Strains


1 Citrobacter amalonaticus ATCC1 25405 feces

2 Citrobacter koseri ATCC 27156 Not Available

3 Citrobacter koseri ATCC BAA-895 clinical specimen - human (infant)

4 Citrobacter braakii ATCC 43162 clinical isolate, California

5 Citrobacter farmeri ATCC 51633 human feces, North Carolina

6 Citrobacter freundii Q Labs2 100813-2A sliced deli meat (turkey)

7 Citrobacter freundii Q Labs 11007-10 Clinical Hospital

8 Citrobacter freundii ATCC 43864 Not Available

9 Citrobacter youngae ATCC 11102 Not Available

10 Enterobacter aerogenes Q Labs 100813-1A Raw vegetable

11 Enterobacter aerogenes ATCC 13048 sputum

12 Enterobacter aerogenes ATCC 35029 Not Available

13 Enterobacter amnigenus ATCC 51816 milk, Minnesota

14 Enterobacter cancerogenus Q Labs 11010-2 Bottled Water

15 Enterobacter cancerogenus Q Labs 11010-1 Bottled Water

16 Enterobacter cloacae Q Labs 100813-3A Soy protein

17 Enterobacter cloacae ATCC 13047 Spinal fluid

18 Enterobacter cloacae NBRC3 13535 Spinal fluid

19 Enterobacter cloacae NBRC 13536 Not Available

20 Enterobacter gergoviae ATCC 33028 Urine, France

21 Hafnia alvei ATCC 51815 Milk

22 Klebsiella oxytoca ATCC 51817 Milk

23 Klebsiella oxytoca ATCC 11686 TBP

24 Klebsiella oxytoca NCTC4 49334 Not Available

25 Klebsiella oxytoca ATCC 700324 Not Available

26 Klebsiella pneumoniae ATCC 11296 Not Available

27 Klebsiella pneumoniae ATCC 4352 Dairy Products (Cow's Milk)

28 Klebsiella pneumoniae ATCC 13440 TBP5

29 Klebsiella pneumoniae Q Labs 11007-7 Raw hamburger

30 Kluyvera intermedia ATCC 33110 Surface Water

31 Pantoea agglomerans ATCC 19552 sewage

32 Rahnella aquatilis ATCC 55046 soil

33 Serratia liquefaciens ATCC 27592 Milk

34 Serratia marcescens Q Labs 11007-1 Bottled Water

35 Serratia marcescens ATCC 14756 Human Isolate

36 Serratia marcescens ATCC 13880 Human Isolate

37 Escherichia coli Q Labs 16160-1 Mixed Frozen Vegetables

38 Escherichia coli Q Labs 16160-2 Frozen Greens

39 Escherichia coli Q Labs 16160-3 Chicken Nuggets

40 Escherichia coli Q Labs 16160-4 Beef Carcass Sponge




44 Escherichia coli Q Labs 16160-8 Fresh Baby Spinach

45 Escherichia coli Q Labs 16160-9 Raw Pork



48 Escherichia coli Q Labs 16160-12 Irrigation Rinse Water

49 Escherichia coli Q Labs 16160-13 Beef Carcass Sponge


1 ATCC-American Type Culture Collection, 2QL- Q Laboratories Culture Collection, 3NBRC-Nite Biological Resource Center,

4NCTC-National Collection Type Cultures, 5TBP – To be provided by 3M

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Table 9: Total Coliform Exclusivity Strains

Number Genus species Source Origin

1 Acinetobacter baumanii ATCC1 19606 Urine

2 Aeromonas viridans QL2 17041-8 Raw Milk Isolate

3 Alcaligenes faecalis ATCC 8750 Not Available

4 Bacillus cereus ATCC 6464 Soil

5 Bacillus subtilis ATCC 6633 Not Available

6 Bordetella bronchiseptica ATCC 10580 Lung of dog

7 Brochothrix thermosphacta ATCC 11509 Animal-derived foodstuff

8 Enterococcus durans ATCC 19432 Not Available

9 Enterococcus faecalis ATCC 29212 Urine

10 Enterococcus faecium ATCC 51559 Clinical isolate

11 Enterococcus hirae ATCC 8043 Not Available

12 Haemophilus influenzae ATCC 19418 Not Available

13 Kurthia gibsonii ATCC 43195 meat

14 Kurthia zopfii ATCC 10538 Not Available

15 Leuconostoc mesenteroides ATCC 8293 fermenting olives

16 Listeria innocua ATCC 33090 cow brain

17 Listeria ivanovii

ATCC BAA-139

washing water

18 Listeria monocytogenes ATCC 7644 Clinical Isolate

19 Listeria seeligeri ATCC 11289 Human feces

20 Listeria welshimeri ATCC 43549 soil

21 Micrococcus luteus ATCC 10240 air

22 Pseudomonas aeruginosa ATCC 27853 Clinical isolate

23 Pseudomonas aeruginosa ATCC 35554 Not Available

24 Pseudomonas fluorescens QL 17041-3 Raw Milk Isolate

25 Staphylococcus hominis ATCC 27844 Human skin

26 Staphylococcus aureus ATCC 6538 human lesion

27 Streptococcus pneumoniae ATCC 6302 Not Available

28 Streptococcus pyogenes ATCC 19615 Pharynx of child following sore

throat.

29 Vibrio parahaemolyticus ATCC 17802 Shirasu food poisoning

30 Vibrio vulnificus QL 021111A Seafood Product 31 Edwardsiella tarda ATCC 15947 Feces, human 32 Morganella morganii ATCC 25829 Human 33 Proteus hauseri ATCC 13315 Human Feces 34 Proteus vulgaris ATCC 6380 Clinical Isolate 35 Providencia rettgeri ATCC 14505 Not Available 36 Providencia stuartii QL 11007-5 Environmental Isolate

37 Salmonella Choleraeusis ATCC 53000

X-ray induced mutant of an equine isolate

38 Salmonella Newport ATCC 6962 food poisoning fatality

39 Salmonella Arizonae QL 11007-4 Veterinary Isolate 40 Yersinia enterocolitica ATCC 49397 clinical specimen

1ATCC-American Type Culture Collection, 2QL – Q Laboratories Culture Collection

Data Intrepreation for Inclusivity and Exclusivity Study

For the inclusivity and exclusivity study, all 100 inclusivity E. coli and total coliform organisms were correctly identified by the 3M Petrifilm Rapid E. coli / Coliform Count Plate method and all 70 exclusivity E. coli and total coliform organisms were correctly excluded by the 3M Petrifilm Rapid E. coli / Coliform Count Plate method, resulting in 100.00% Postive Agreement (PA) and a 0.00% Negative Deviation (ND).

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The strains provided by 3M were strictly used in the Inclusivity portion of the study and not used in the development of the product.

4 Interlaboratory Study

The interlaboratory study (ILS) was organized and data evaluated according to the details provided in ISO 16140-2:2016.

4.1 Organization

The ILS was performed using 9 collaborators from 8 organizations representing 5 different countries. The food item used in the evaluation was dry dog kibble. The food item was obtained from a local retailer and screened for the presence of naturally occurring E. coli and coliform by the ISO 16649-2 and ISO 4832 reference methods. No natural contamination was observed so four separate levels of contamination were targeted for the evaluation: uninoculated, 0 CFU/g; low, 10-100 CFU/g; medium, 100-1,000 CFU/g; and high 1,000-10,000 CFU/g. To obtain the required contamination levels, bulk lots of the matrix were artificially contaminated with a lyophilized culture of Escherichia coli [Q Laboratories (QL) isolate 11007-8 (origin – beef hide)] and Klebsiella pneumonia [Q Laboratories (QL) isolate 11007-7 (origin – raw hamburger)] at each target contamination level. Both organisms were included in order to determine the target total coliform contamination level. Two replicate samples from each of the four contamination levels were analyzed by both the candidate and reference methods in a paired study design. Preparation of the Inocula and Test Portions The E. coli and K. pneumonia isolates used in this evaluation was lyophilized prior to inoculation. The cultures were first propagated onto Tryptic Soy Agar with 5% Sheep Blood (SBA) from a Q Laboratories frozen stock culture stored at -70°C. To prepare the cultures for lyophilization, a single, well isolated colony from SBA was transferred into BWP (ISO) and incubated at 37 ± 2 oC for 18-24 hours. The cultures were diluted in a sterile cryoprotectant, reconstituted 10% non-fat dry milk (NFDM), and freeze dried for 48-72 hours. A bulk lot of the test matrix was inoculated with each culture at a high level expected to yield all positive results. An aliquot of the high-level inoculum was further mixed with uninoculated powdered infant formula to produce the low-level inoculum. After inoculation, the matrix was held for a minimum of 2 weeks at ambient temperature (20 - 25oC). The inoculated test product was packaged into separate 25 g samples in sterile Whirl-Pak® bags and shipped to the collaborators.

Test Portion Distribution All samples were labeled with a randomized, blind-coded 3 digit number affixed to the sample container. Nine (9) participants from 8 separate locations participated. Test portions were shipped in leak-proof insulated containers via overnight delivery according to the Category B Dangerous Goods shipment regulations set forth by

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International Air Transport Association (IATA). Test portions were shipped at ambient temperatures (20-25oC). Upon receipt, samples were held at ambient temperature until analysis was initiated. In addition to each of the test portions, collaborators received a test portion for the matrix labeled as Aerobic Plate Count (APC), to determine total background count in the matrix. The APC background screen samples were prepared from the bulk lot of test matrix, prior to inoculation. Additionally, a temperature probe was included in the shipment. Participants were instructed to submit the data from the temperature probe upon receipt of the shipment.

Table 6: Sample receipt data for ILS samples

Collaborator Temperature

(°C)

1 20.7

2 21.5

3 21.4

4 23.2

5 20.9

6 24.8

7 23.6

8 24.5

9 20.2

4.2 Calculations and interpretation of data The data from the collaborative trial were calculated and interpreted according to

section 6.2.3 of ISO 16140-2:2016 using the freely available Excel® spreadsheet

(http://standards.iso.org/iso/16140).

The log transformed data for the coliform and E. coli results of the alternative method

and reference method are presented in Tables 7 and 8, respectively. The Accuracy

profile graph is shown in Figure 9A and 9B.

http://standards.iso.org/iso/16140

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Table 7: Summary of the results of the ILS: 3M Petrifilm REC- Coliform 37 ± 1oC

Collaborators Level Reference method Alternative method

ISO 4832 3M Petrifilm REC

1 Uninoculated <1.000 <1.000 <1.000 <1.000

2 Uninoculated <1.000 <1.000 <1.000 <1.000

3 Uninoculated <1.000 <1.000 <1.000 <1.000

4 Uninoculated <1.000 <1.000 <1.000 <1.000

5 Uninoculated <1.000 <1.000 <1.000 <1.000

6 Uninoculated <1.000 <1.000 <1.000 <1.000

7 Uninoculated <1.000 <1.000 <1.000 <1.000

8 Uninoculated <1.000 <1.000 <1.000 <1.000

9 Uninoculated <1.000 <1.000 <1.000 <1.000

1 Low 1.708 1.613 2.004 1.785

2 Low 1.613 1.041 1.908 1.322

3 Low 1.708 1.708 1.613 1.041

4 Low 1.322 1.785 1.041 1.041

5 Low 1.785 1.613 1.785 1.851

6 Low 1.908 1.908 1.613 1.908

7 Low 1.785 2.004 1.322 1.959

8 Low 2.004 1.613 1.491 1.785

9 Low 1.491 1.851 1.613 2.004

1 Medium 2.179 2.004 2.344 2.233

2 Medium 2.603 2.654 2.545 2.400

3 Medium 2.449 2.592 2.520 2.682

4 Medium 2.603 2.741 2.344 2.569

5 Medium 2.673 2.700 2.400 2.520

6 Medium 2.400 2.303 2.581 2.624

7 Medium 2.779 2.930 2.741 2.876

8 Medium 2.581 2.700 2.820 2.846

9 Medium 2.634 2.493 2.700 2.741

1 High 3.179 3.344 3.004 3.083

2 High 3.303 3.258 3.179 3.344

3 High 3.581 3.545 3.382 3.400

4 High 3.464 3.400 3.258 3.281

5 High 3.233 3.400 3.433 3.592

6 High 3.258 3.179 3.400 3.507

7 High 3.233 3.433 3.507 3.344

8 High 3.344 3.479 3.464 3.464

9 High 3.400 3.303 3.533 3.281

77/92

Table 8: Summary of the results of the ILS: 3M Petrifilm REC – E. coli 37 ± 1oC


ISO 16449-2 3M Petrifilm REC

1 Uninoculated <1.000 <1.000 <1.000 <1.000

2 Uninoculated <1.000 <1.000 <1.000 <1.000

3 Uninoculated <1.000 <1.000 <1.000 <1.000

4 Uninoculated <1.000 <1.000 <1.000 <1.000

5 Uninoculated <1.000 <1.000 <1.000 <1.000

6 Uninoculated <1.000 <1.000 <1.000 <1.000

7 Uninoculated <1.000 <1.000 <1.000 <1.000

8 Uninoculated <1.000 <1.000 <1.000 <1.000

9 Uninoculated <1.000 <1.000 <1.000 <1.000

1 Low 1.613 1.322 1.613 1.708

2 Low 1.491 1.041 1.908 1.322

3 Low 1.708 1.322 1.491 1.041

4 Low 1.322 1.613 1.041 1.041

5 Low 1.785 1.613 1.613 1.041

6 Low 1.785 1.708 1.491 1.491

7 Low 1.322 1.708 1.322 1.785

8 Low 1.851 1.613 1.041 1.613

9 Low 1.322 1.613 1.491 1.491

1 Medium 2.149 2.004 2.083 2.303

2 Medium 2.364 2.344 2.258 2.004

3 Medium 2.303 2.179 2.045 1.959

4 Medium 2.581 2.417 2.149 2.417

5 Medium 2.520 2.464 2.449 2.179

6 Medium 2.149 2.258 2.303 2.303

7 Medium 2.281 2.520 2.417 2.520

8 Medium 2.233 2.258 2.479 2.507

9 Medium 2.303 2.004 2.464 2.400

1 High 3.083 3.303 3.004 2.908

2 High 3.258 2.959 3.004 3.179

3 High 3.470 3.479 3.281 3.083

4 High 3.303 3.382 3.004 3.083

5 High 3.507 3.083 3.179 3.004

6 High 3.004 2.785 3.149 3.004

7 High 3.207 3.400 3.479 3.258

8 High 3.045 3.083 3.233 3.149

9 High 3.004 3.281 3.303 3.207

78/92

Table 9: Summary of the results of the ILS: 3M Petrifilm REC – E. coli 42 ± 1oC


ISO 16449-2 3M Petrifilm REC

1 Uninoculated <1.000 <1.000 <1.000 <1.000

2 Uninoculated <1.000 <1.000 <1.000 <1.000

3 Uninoculated <1.000 <1.000 <1.000 <1.000

4 Uninoculated <1.000 <1.000 <1.000 <1.000

5 Uninoculated <1.000 <1.000 <1.000 <1.000

6 Uninoculated <1.000 <1.000 <1.000 <1.000

7 Uninoculated <1.000 <1.000 <1.000 <1.000

8 Uninoculated <1.000 <1.000 <1.000 <1.000

9 Uninoculated <1.000 <1.000 <1.000 <1.000

1 Low 1.613 1.322 1.322 1.041

2 Low 1.491 1.041 1.708 1.041

3 Low 1.708 1.322 1.322 1.041

4 Low 1.322 1.613 1.491 1.041

5 Low 1.785 1.613 1.491 1.491

6 Low 1.785 1.708 1.491 1.708

7 Low 1.322 1.708 1.041 1.613

8 Low 1.851 1.613 1.322 1.708

9 Low 1.322 1.613 1.613 1.708

1 Medium 2.149 2.004 2.303 2.233

2 Medium 2.364 2.344 2.520 2.303

3 Medium 2.303 2.179 2.083 2.258

4 Medium 2.581 2.417 2.233 2.479

5 Medium 2.520 2.464 2.449 2.233

6 Medium 2.149 2.258 2.303 2.004

7 Medium 2.281 2.520 2.382 2.479

8 Medium 2.233 2.258 2.464 2.520

9 Medium 2.303 2.004 2.507 2.400

1 High 3.083 3.303 3.045 2.908

2 High 3.258 2.959 3.004 3.149

3 High 3.470 3.479 3.083 2.908

4 High 3.303 3.382 3.083 3.281

5 High 3.507 3.083 3.233 3.233

6 High 3.004 2.785 3.303 3.149

7 High 3.207 3.400 3.281 3.045

8 High 3.045 3.083 3.083 3.449

9 High 3.004 3.281 3.149 3.004

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Figure 9A: Accuracy profile of the alternative method (Coliform 37 ± 1oC) in the ILS

-0,8

-0,6

-0,4

-0,2

0,0

0,2

0,4

0,6

0,8

1,5 2,0 2,5 3,0 3,5

Accura

cy (

diffe

rence o

f L

og)

Levels Log(cfu/g)

Bias

Relative Lower TI limit (beta = 80%)

Relative Upper TI limit (beta = 80%)

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Figure 9B: Accuracy profile of the alternative method (E. coli 37 ± 1oC ) in the ILS

-0,8

-0,6

-0,4

-0,2

0,0

0,2

0,4

0,6

0,8

1,5 2,0 2,5 3,0 3,5

Accura

cy (

diffe

rence o

f L

og)

Levels Log(cfu/g)

Bias



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Figure 9C: Accuracy profile of the alternative method (E. coli 42 ± 1oC ) in the ILS

The ILS statistical analysis is presented in Tables 10 -12. Based on data presented in these tables, the repeatibility standard deviations (Sr) and between-labs standard devaiations (SL) indicate similar results between the alternative and reference method. The reproducibility standard deviation (SR) indicated slightly better results with the alternative method than the reference method.

According to the ISO 16140-2:2016 standard, if any of the values of the β-ETI fall outside of the ±0.5log AL then a further calculation is required. None of the β-ETI values fell outside the range, indicating no additonal calculations were required. Based on the data obtained within these studies, the alternative method is considered equivalent to the reference methods.

-0,8

-0,6

-0,4

-0,2

0,0

0,2

0,4

0,6

0,8

1,5 2,0 2,5 3,0 3,5

Accura

cy (

diffe

rence o

f L

og)

Levels Log(cfu/g)

Bias



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Table 10: Statistical analysis of the ILS data (3M Petrifilm REC – Coliform 37 ± 1oC)

Accuracy profile 0.5

Study Name

Date

Coordinator TRUE

Tolerance probability (beta) 80% 80% 80%

Acceptability limit in log (lambda) 0.68 0.68 0.68

Alternative method Reference method

Levels Low Medium High Low Medium HighTarget value 1.682 2.555 3.350

Number of participants (K) 9 9 9 9 9 9

Average for alternative method 1.601 2.581 3.357 1.682 2.555 3.350

Repeatability standard deviation (sr) 0.293 0.091 0.095 0.233 0.088 0.087

Between-labs standard deviation (sL) 0.194 0.170 0.129 0.084 0.213 0.085

Reproducibility standard deviation (sR) 0.351 0.193 0.160 0.247 0.231 0.122

Corrected number of dof 15.018 9.981 11.299 16.494 9.270 13.112

Coverage factor 1.388 1.439 1.422

Interpolated Student t 1.341 1.372 1.361

Tolerance interval standard deviation 0.3636 0.2021 0.1677

Lower TI limit 1.114 2.304 3.128

Upper TI limit 2.089 2.859 3.585

Bias -0.080 0.026 0.007

Relative Lower TI limit (beta = 80%) -0.568 -0.251 -0.222 TRUE

Relative Upper TI limit (beta = 80%) 0.407 0.303 0.235 FALSE

Lower Acceptability Limit -0.68 -0.68 -0.68

Upper Acceptability Limit 0.68 0.68 0.68

New acceptability limits may be based on reference method pooled variancePooled repro standard dev of reference 0.207

3M Petrifilm Rapid E.coli/Coliform Count Plate

Oct-18

Q Laboratories

Select ALL blue lines to draw the accuracy profile as illustrated in the worksheet "Graph Profile"

Application of clause 6.2.3 Step 8: If any of the values for the β-ETI fall outside the

acceptability limits, calculate the pooled average reproducibility standard deviation of the reference

method.Step 9: Calculate new acceptability limits as a function

of this standard deviation.

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Table 11: Statistical analysis of the ILS data (3M Petrifilm REC – E. coli 37 ± 1oC)


Study Name

Date

Coordinator TRUE














Lower TI limit 0.996 2.026 2.933

Upper TI limit 1.801 2.551 3.340

Bias -0.107 -0.018 -0.043







Oct-18

Q Laboratories

Select ALL blue lines to drawthe accuracy profile as illustrated in the worksheet "Graph Profile"

Application of clause 6.2.3 Step 8: If any of the values for the β-ETI fall outside

the acceptability limits, calculate the pooled average reproducibility standard deviation of the reference

method.Step 9: Calculate new acceptability limits as a

function of this standard deviation.

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Table 12: Statistical analysis of the ILS data (3M Petrifilm REC – E. coli 42 ± 1oC)


Study Name

Date

Coordinator TRUE














Lower TI limit 0.986 2.128 2.930

Upper TI limit 1.771 2.552 3.329

Bias -0.127 0.033 -0.050







Oct-18

Q Laboratories

Select ALL blue lines to draw the accuracy profile as illustrated in the worksheet "Graph Profile"

Application of clause 6.2.3 Step 8: If any of the values for the β-ETI fall outside the

acceptability limits, calculate the pooled average reproducibility standard deviation of the reference

method.Step 9: Calculate new acceptability limits as a function

of this standard deviation.

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5 References

1. International Standard: ISO 16140-1 – Microbiology of the food chain – Method validation. Part 1: Vocabulary, First Edition, 2016-06-15.

2. International Standard: ISO 16140-2 – Microbiology of the food chain – Method validation. Part 2: Protocol for the validation of alternative (proprietary) methods against a reference method. First Edition, 2016-06-15.

3. International Standard: ISO 4832:2006 – Microbiology of food and animal feeding stuffs – Horizontal method for the enumeration of coliforms – Colony-count technique. Third Edition, 2006-02.

4. International Standard: ISO 16649-2:2001 – Microbiology of food and animal feeding stuffs – Horizontal method for the enumeration of β-glucuronidase-positive Escherichia coli - Part 2: Colony-count technique at 44°C using 5-bromo-4-chloro-3-indolyl-β-D-glucuronide. First Edition, 2001-04.

5. International Standard: ISO 6887 (All parts) – Microbiology of food and animal feeding stuffs – Preparation of test samples, initial suspension and decimal dilutions for microbiological examination. First Editions.

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6 ANNEX A: Flow Diagram of the ISO 4832 Reference Method

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7 ANNEX B: Flow Diagram of the ISO 16649-2:2001 Reference Method

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8 ANNEX C: Flow Diagram of the 3M Petrifilm Rapid E. coli and Coliform Count Plate Alternative Method

9 ANNEX D: Kit Insert for 3M Petrifilm Rapid E. coli / Coliform Count Plate

See the attached document.

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10 ANNEX E: Summary of Artificial Contamination

Trueness Study

Category Type Sample # Item

Target

Inoculum

Level

Inoculating

Organisms Origin Stressing Hold time

Lo

g1

0

Inju

ry

Meat Products Breaded Meat

QL017 Chicken Nuggets 1 x 103 Klebsiella pneumonia

QL 11007-7 & E.

coli QL 16160-4

Meat &

Chicken

Nuggets

Heat

50 ºC for

10 mins

0.62 QL018 Spicy Chicken

Nuggets 1 x 104

QL019 BBQ Chicken Wings 1 x 105

Dairy and Egg

Products

Pasteurized Dairy

Products

QL035 Butter 1 x 103 Cronobacter

condimenti

DSM 27966 & E.

coli

QL 11010-2

Infant Formula

& Bottled

Water

5⁰C for 48-72

hours QL036 Ricotta 1 x 103

QL037 Whipping Cream 1 x 103

Desserts & Egg

Products

QL056 Rice Pudding 1 x 102 Hafnia alvei

ATCC 51815 & E.

coli

QL 16154-7

Milk,

Minnesota &

Flour

5⁰C for 48-72

hours

QL057 Cream Puffs 1 x 102


Raw Products

QL067 Cod 1 x 105

Serratia marcescens

ATCC 8100 & E. coli

QL 16154-2

Milk, Cork

Ireland & Raw

Ground Beef

5⁰C for 48-72

hours

QL068 Pollock 1 x 105

QL069 Shrimp 1 x 105

QL071 Catfish 1 x 103

QL072 Trout 1 x 104

Marinated Smoked

Items

QL081 Lemon Pepper

Scallops 1 x 103

Klebsiella pneumonia

QL 11007-7 & E.

coli QL 16160-8

Meat &

Chicken

Carcass Rinse

Heat

50ºC for

10 mins

0.56

QL082 Smoky Bourbon

Salmon 1 x 103

90/92

Cooked Products

QL091 Cooked Shrimp 1 x 103 Citrobacter freundii

QL 100813-2A &

E. coli QL 16160-10

Sliced deli

turkey meat &

Raw Pork

Heat

50ºC for

10 mins

0.67 QL093 Lobster Cakes 1 x 102

QL094 Salmon Burgers 1 x 102

Vegetable

Products

Pre-Cooked

Vegetables

QL111 Frozen Lima Beans 1 x 103

Serratia marcescens

ATCC 8100 & E. coli

QL 16160-2

Milk, Cork

Ireland &

Mixed Frozen

Vegetables

-20⁰C for 2

weeks

QL112 Frozen Green Beans 1 x 103

QL113 Frozen Carrots 1 x 103

QL114 Frozen Onions 1 x 103

QL115 Frozen Corn 1 x 103

Ready to Eat and Ready to

Reheat

Cold Ready Meals

QL130 Roast Beef Pinwheel 1 x 104

Citrobacter freundii

QL 100813-2A &

E. coli QL 16154-3

Sliced deli

turkey meat &

Beef Trim

5⁰C for 48-72

hours QL131

Ham and Turkey

Sandwich 1 x 104

QL132 Uncrustable PB + J 1 x 104

Pasta

QL144 Macaroni & Cheese 1 x 103

Klebsiella pneumonia

QL 11007-7 & E. coli QL 16154-13

Meat &

Potable Water

Heat

50ºC for

10 mins

0.59

QL145 Lasagna 1 x 103

QL146 Cheese Ravioli 1 x 102

QL147 Lentil Pasta with

Cheddar Sauce 1 x 102

Animal Feed

Products

Finished Products

for Domestics QL160

Temptations Cat

Treats 1 x 103

Serratia marcescens

ATCC 8100 & E. coli

QL 16154-2

(Lyophilized)

Milk, Cork

Ireland & Raw

Ground Beef

Ambient for 2

weeks

Ingredients for

Domestic/Livesto

ck

QL176 Soybean Meal 1 x 102

Rahnella aqualitis

ATCC 55046 & E.

coli QL 16154-7

(Lyophilized)

Soil & Flour Ambient for

2 weeks

QL177 Rolled Oats 1 x 102

QL178 Yellow Cornmeal 1 x 103

QL179 Pearled Barley 1 x 103

QL181 Long Grain White

Rice 1 x 103

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Accuracy Study

Category Type Item

Target

Inoculum

Level

Inoculating

Organism Origin Spiking Seeding

%

Injury

Meat Products

(Combined

category raw/RTC

meats and poultry)

Dairy and Egg

Products

Raw Fresh

Meats Minced Meat

~1.0 x 102 Klebsiella pneumoniae

ATCC 13882 & E. coli QL 16154-2

Water & Raw

Ground Beef

5°C 48 to 72

Hours ~1.0 x 103

~1.0 x 105

Breaded Meats Raw Frozen Chicken

Wings

~1.0 x 102 Klebsiella pneumonia ATCC 11296 &

E. coli QL 16154-12

Water & Chicken

Carcass Rinsate -20°C 2 weeks ~1.0 x 103

~1.0 x 105


Raw Dairy

Products Raw Cow’s Milk

~1.0 x 102 Hafnia alvei ATCC 51815 &

E. coli QL 16160-4

Milk & Chicken

Nuggets

5°C 48 to 72 Hours

~1.0 x 103

~1.0 x 105

Egg Products

(heat processed)

with additives

Whole Liquid Eggs

~1.0 x 102 Hafnia alvei QL 102313 &

E. coli QL 11010-2

Milk & Bottled

Water

5°C 48 to 72 Hours

~1.0 x 103

~1.0 x 105


Raw Products Tuna Sushi

~1.0 x 102 Enterobacter amnigenus

ATCC 51816 & E. coli QL 16160-16

Milk, Minnesota

& Sprouts

5°C 48 to 72

Hours ~1.0 x 103

~1.0 x 105

Smoked

Products Smoked Salmon

~1.0 x 102 Enterobacter amnigenus

QL 112413-1 & E. coli 3M REC8

Milk & Food

Isolate

5°C 48 to 72 Hours

~1.0 x 103

~1.0 x 105

Vegetable Products

Raw Vegetable

products Bunched Spinach

~1.0 x 102 Klebsiella oxytoca ATCC 13182 &

E. coli 3M REC19

Pharyngeal

tonsil & Not

Available

5°C 48 to 72

Hours ~1.0 x 103

~1.0 x 105

Heat Processed

Vegetable Juices Pasteurized Carrot Juice

~1.0 x 102 Klebsiella oxytoca ATCC 700324 &

E. coli QL 16160-9

Not Avaliable &

Fresh Baby

Spinach

Heat

50ºC for

10 mins

0.63 ~1.0 x 103

~1.0 x 105

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Accuracy Study

Category Type Item

Target

Inoculum

Level

Inoculating


%

Injury

Accuracy Study

Category Type Item

Target

Inoculum

Level

Inoculating


%

Injury

Ready to Eat and Ready to Reheat

Cold Ready

Meals

Ready-made

Sandwiches

~1.0 x 102

Kluyvera intermedia ATCC 33110 &

E. coli 3M REC17

Surface Water &

Not Avaliable

5°C 48 to 72

Hours

~1.0 x 103

~1.0 x 105

Mayonnaise

based deli

salads with raw

ingredients

Raw Vegetable Salad

with Dressing

~1.0 x 102 Kluyvera intermedia

QL 081215 & E. coli QL 16160-12

Water &

Irrigation Rinsate

Water

5°C 48 to 72

Hours ~1.0 x 103

~1.0 x 105

Animal Feed

Finished

Products fro

Livestock

Chicken Feed

~1.0 x 102 Enterobacter cloacae NBRC 13536 &

E. coli QL 16160-5

Not Avaliable &

Flavored

Seasoning

Ambient 2 weeks ~1.0 x 103

~1.0 x 105

Plant Origin

Ingredients Soybean Meal

~1.0 x 102 Enterobacter cloacae NBRC 13535 &

E. coli 3M REC11

Spinal Fluid &

Food Isolate Ambient 2 weeks ~1.0 x 103

~1.0 x 105

Samples from the

Industrial

Production

Environment

Surface Samples

Stainless Steel

Environmental Spones

~1.0 x 102 Enterobacter gergoviae

QL 123009-1 & E. coli 3M REC1

Clinical &

Mozzerella

Cheese

Ambient

16 to 24 Hours ~1.0 x 103

~1.0 x 105

Sealed Concrete

Environmental Sponges

~1.0 x 102 Enterobacter gergoviae

ATCC 33028 & E. coli QL 16160-10

Urine, France &

Raw Pork

Ambient 16 to 24 Hours

~1.0 x 103

~1.0 x 105

MicroVal Study 2017LR76 - Betty Blocks€¦ · The 3M Petrifilm Rapid E. coli / Coliform Count...

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