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Annex D-2: CFIA risk-based Shiga-toxin-producing E. coli verification sampling of beef trimmings for abattoirs eligible for export to the USA

1. Purpose and scope

This verification sampling has been designed by the Canadian Food Inspection Agency (CFIA) for abattoirs producing raw beef manufacturing trimmings for the export to the USA. The participation in this program is mandatory for abattoirs in order to maintain their eligibility for export to the USA. The samples will be collected by the operators under the supervision of CFIA veterinarian-in-charge (VIC) and submitted by the operators to private laboratories for the analysis of Shiga toxins-producing Escherichia coli (STEC) O26, O103, O111, O121, O45, O145.

2. Products targeted for sampling

Products targeted for sampling under this program are raw boneless beef manufacturing trimmings, that is to say, pieces of meat remaining after steaks, roasts and other intact cuts are removed.

3. Frequency of sampling

The normal sampling frequency under this new program will match the frequency under the existing monitoring sampling plan M218 that was determined on the basis of risk considering the following four factors:

  1. production volume
  2. compliance history
  3. E. coli O157:H7 robust testing employed by the establishment, and
  4. the seasonal variations in STEC prevalence

On the basis of production volume the establishments have been divided into four categories (as defined in M218):

The positive E. coli O157:H7 results obtained over the last 120 days under the CFIA monitoring program for E. coli O157:H7 (M201) should be used for the assessment of compliance history. The O157:H7 data is used since the sources of contamination for all seven STEC are thought to be similar and the interventions employed by operators under their HACCP system are believed to exert a similar protection against O157 and the six other STEC.

The number of samples to be collected under the plan will be based on Table 1.

Table 1: Risk-based sampling frequency for each federally registered establishment
Establishment size (production volume per year) Normal frequency Table note 1 Enhanced frequency Table note 3
Sampling frequency October to March Sampling frequency April to September Table note 2 Number of samples per year per establishment Sampling frequency October to March Sampling frequency April to September Table note 2
(<25K kg)
1 per month 1 per month 12 2 per month 2 per month
(25K to 400K kg)
1 per month 3 per 2 months 15 2 per month 3 per month
(400K to 40M kg)
3 per 2 months 2 per month 21 3 per month 4 per month
Extra large
(>40M kg)
2 per month 4 per month 36 4 per month 8 per month

Table Notes

Table Note 1

Generally, all establishments will be sampled at a normal frequency.

Return to table note 1  referrer

Table Note 2

Includes additional samples taken at each establishment during the high prevalence period of April to September.

Return to table note 2  referrer

Table Note 3

Enhanced frequency - A compliance history including a positive E. coli O157 result from M218 testing of precursor material or M201 testing of the product downstream will be taken into account when placing an establishment on enhanced frequency of testing for the next 120 days. As well High Event Period (HEP) will be taken into consideration when determining frequency of sampling. HEP is a situation in which an establishment experiences a high number or rate of positive results for E. coli O157. Such a decision will be made by the Area Program Specialist.

Return to table note 3  referrer

4. Procedure for collecting and submitting samples for the analysis

Operators are responsible for collection of samples from the lot that has been selected by the CFIA inspector under sampling plan M218. The operator's written procedure for the collection and submission of samples for analysis will be approved by the VIC. The following steps are to be followed:

4.1. A lot of beef trimmings assembled for sampling under M218 will be concurrently sampled by the operator. A designated employee trained in the N60 sampling procedure will collect samples according to plan M218 at the time when sampling is accomplished by a CFIA inspector and under his or her supervision. The inspector notifies the establishment management 24 hours before the planned sampling. This will provide establishments enough time to hold the entire lot from which the samples are collected pending the result of analysis. The CFIA strongly recommends holding the entire lot pending the result of analysis.

4.2 A random sample will be collected after a lot has been assembled as per the operator's lot definition.

4.3 A sample will be collected aseptically by using N60 method as described in section 5. N60 method of sample collection.

4.4 Immediately after sampling the inspector will check and record the product temperature from one of the sampled containers (do not take the temperature of the sample). When the temperature of the product is higher than 4°C (40°F), the bag containing the sample will be placed in a cooler to chill before shipping.

4.5 The CFIA inspector will oversee the preparation of sample for shipping as per CFIA standards (tamper evident) using sufficient ice packs and ensure that the sample integrity will be maintained from the collection to shipping.

5. N60 method of sample collection

Samples of beef trimmings will be collected using the N60 method as described below.

5.1 Use Table 2 below for sampling. If a specific production lot is composed of more than 5 containers of raw beef trimmings, randomly select 5 containers for sampling.

Table 2: Number of pieces to be collected per combo (container) when a lot consists of 5 or less combos (containers)
Number of containers in a specific production lot Number of sample pieces to select from each combo (container)
5 12
4 15
3 20
2 30
1 60

5.2 Aseptically collect the appropriate number of pieces as indicated in Table 2. The sample is comprised of 60 pieces.

5.3 Using a sanitized knife, cut off a thin slice that is approximately 50 cm2 for each of the 60 pieces, for example: 10 cm (4") × 5 cm (2") × 0.3 cm (1/8"). Collect as much of the beef trimmings outer surface as possible. The priority is to collect samples from pieces of product taken from the original surface of the beef carcass.

5.4 Place the sampled slices into the sterile sampling bag. Weigh the sample to ensure that approximately 1 kg (2 pounds) was collected.

6. Lot definition

A lot is defined as for the sampling under M218. It can consist of 1-5 combos.

7. Target organisms

The samples will be analysed for the presence of six STECs, namely O26, O103, O111, O121, O45, O145. Some Food Safety and Inspection Services (FSIS) testing methods include the detection of O157. If this is a method of choice, the results obtained on a sample will also include the O157 serotype.

8. Method of analysis

8.1 Any test method that has been granted a No Objection Letter (NOL) from the United States Department of Agriculture (USDA) - Food Safety and Inspection Service (FSIS) can be used for screening for the presence of STEC.

The updated list of screening methods is published on the FSIS Microbiology Laboratory Guidebook.

8.2 The FSIS cultural method described under the USDA-FSIS Microbiology Laboratory Guidebook 5B.03, as well as other confirmation methods that have been granted a NOL by FSIS (available on the FSIS website) are acceptable for the cultural confirmation of presumptive positive results (isolation and identification of STEC of interest from the presumptive positive broth).

8.3 It is the operator's responsibility to verify before shipping samples for the analysis that the laboratory:

  1. has on their scope of accreditation any of the testing methods that obtained a NOL from the FSIS for analysing raw beef trimmings for the STECs of interest
  2. or, has initiated the application process with the Standards Council of Canada or the Canadian Association for Laboratory Accreditation (CALA) to add any of these methods to their scope of accreditation and the laboratory has previously been accredited for at least one testing method for other micro-organisms (for example: Salmonella, Listeria monocytogenes, E. coli, etc.) in food.

9. Reporting of the results

Operators are required to convey Annex D-2.1: STEC in Beef Trimmings Verification Sampling Template and Annex D-2.2: Instructions for Completion of Shiga Toxins-Producing Escherichia coli (STEC) in Beef Trimmings Verification Sampling Template to the private laboratories they contract testing to, which include specific instructions for reporting test results to the CFIA.

All results obtained under this program, that is to say, negative, presumptive positive and culturally confirmed positives will be reported by the operator's contracted laboratory to the CFIA's Food Safety Division for the maintenance of a database and the analysis of results. The results will be entered into the template by the private laboratory and sent along with the supportive record/certificate of analysis to the following address Positive (present) results should be classified as high priority or urgent, and submitted within 1 business day. Negative (absent) results may be combined into one template and submitted within the last week of each month.

10. Follow-up on positive results

10.1 Trimmings that tested presumptive positive (that do not confirm as negative if confirmation methods are performed), and/or confirmed positive for any of the seven STEC of interest must not be exported to the USA also will not be released into domestic market.

10.2 The operator may choose to confirm culturally the presumptive positive results or treat presumptive positives as confirmed positives and as such can only dispose of these trimmings as per the guidance for E. coli O157:H7 provided in Preventive controls for E. coli O157/NM in raw beef products. Presumptive positive results that confirm culturally to be negative (that is to say, the screen method gave a false positive result), may be considered suitable for sale on the market place.

The operator will notify the CFIA inspector within 24 hours of negative and presumptive positive results. Upon receiving a presumptive positive result, the inspector will immediately ensure that the implicated trimmings are on hold. In the case that the implicated trimmings were distributed, the inspector will immediately notify the Area Meat Program Specialist and Area Recall Coordinator.

For any further information on this sampling project contact the CFIA (Contact CFIA online).

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