Annex D-2: CFIA risk-based Shiga toxin-producing E. coli verification sampling of raw beef products in Canadian beef establishments eligible for export to the USA

1. Purpose and scope

Canadian Food Inspection Agency (CFIA) risk-based Shiga toxin-producing E. coli (STEC) verification sampling has been designed by the CFIA for Canadian federally inspected beef establishments producing raw beef products (see section 2) for the export to the USA. The participation in this program is mandatory for abattoirs and processing establishments in order to maintain their eligibility for export of beef to the USA. In case the abattoir does not have a cutting room, the processing establishment that processes the carcasses from this establishment must perform the verification sampling.

The samples will be collected by the operators of the establishment under the supervision of CFIA and submitted by the operators to private laboratories for the analysis of Shiga toxins-producing Escherichia coli (STEC) O26, O103, O111, O121, O45, O145.

The CFIA signing official shall review the test results for the product being exported to the United States to ensure that the results are satisfactory, and the correct methodology was used before signing the health certificate.

2. Products targeted for sampling

Products targeted for sampling under this program are:

raw boneless beef manufacturing trimmings produced from cattle slaughtered onsite (i.e., pieces of meat remaining after steaks, roasts and other intact cuts are removed).
raw beef bench trimmings from carcasses of cattle not slaughtered onsite,
raw ground beef, and
raw ground beef components as follows: head meat, cheek meat, weasand (esophagus) meat, meat from Advance Meat Recovery (AMR), Partially Defatted Chopped Beef (PDCB), Finely textured beef and heart meat.

Only finished products (those shipped from the establishment) should be targeted for sampling. For example, if the establishment makes beef manufacturing trimmings but uses all of those trimming in-house to make ground beef, beef manufacturing trimmings should NOT be targeted.

3. Frequency of sampling

The normal sampling frequency under this program will match the frequency under the existing monitoring sampling plan M218 that is determined on the basis of risk considering the following 4 factors:

production volume
compliance history
E. coli O157:H7 robust testing employed by the establishment and
the seasonal variations in STEC prevalence
On the basis of production volume the establishments have been divided into 4 categories (as defined in M218): extra large, large, medium and small. 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 E. coli O157:H7 data is used since the sources of contamination for all 7 STEC are thought to be similar and the interventions employed by operators under their Hazard Analysis and Critical Control Point (HACCP) system are believed to exert a similar protection against E. coli O157:H7 and the 6 other STEC.

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

Table 1: Risk-based sampling frequency for each federally inspected establishment for raw beef products for STEC
Establishment size (production volume per year)	Normal frequency ^{Table note 1}			Enhanced frequency ^{Table note 3}
Establishment size (production volume per year)	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}
Small (<25K kg)	1 per month	1 per month	12	2 per month	2 per month
Medium (25K to 400K kg)	1 per month	3 per 2 months	15	2 per month	3 per month
Large (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 – An establishment may be placed on an enhanced testing frequency – A for 120 days if its compliance history includes:

a positive E. coli O157:H7 result from M218 testing of precursor material or
a positive result from M201 testing of downstream product. In addition, High Event Period (HEP) occurs when an establishment experiences a high number or rate of positive E. coli O157:H7 results. Decisions regarding enhanced sampling frequency, including those based on HEPs, will be made by the Area Operations Specialist responsible for Export.

Return to table note 3 referrer

4. Procedure for collecting and submitting samples for analysis

Operators are to sample, raw beef manufacturing trimmings, raw beef bench trimmings, raw ground beef and raw beef components other than trim separately. When the establishment produces multiple types of trimmings or multiple types of raw ground beef components other than trim, operators are to target each type under their sampling. However, for a given sampling event, operators are to collect sample of only one type of trimming or one type of raw ground beef component other than trim. The intent is that, through random selection, all eligible products and their types that are produced at the establishment and are subject to STEC sampling, will be selected over time.

If applicable, operators will collect samples from the lot that has been selected by the CFIA inspector under sampling plan M218. If the product that is due for operator sampling is different than the CFIA sampling under M218 (for e.g. operator has to sample ground beef), then operator will take sample under CFIA supervision from another lot selected by the CFIA. The operator's written procedure for the collection and submission of samples for analysis will be reviewed and accepted by the CFIA. The following steps are to be followed:

4.1. 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.

4.2 A random sample will be collected under CFIA supervision after a lot has been assembled (see lot definition below).

4.3 A sample will be collected aseptically by using the appropriate sampling method as described in section 5.

4.4 Immediately after sampling the inspector will check and record the product temperature from 1 of the sampled containers (combos) (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.

4.6 CFIA requires establishments to hold or maintain control of raw beef products that has tested for STEC pending negative results.

5. Sample collection

A licence holder may either use one of the sampling methods listed below or may use an alternate sampling procedure if the alternate sampling procedure and associated test methodology is as/or more rigorous than the procedure listed below and has been reviewed and accepted by the CFIA prior to its use.

5.1 N60 method of sample collection

For beef products that are amenable to excision sampling (for example beef trimmings, ground beef components other than trim (like head meat, cheek meat, weasand (esophagus) meat, heart meat)) will be collected using the N60 method as described below.

5.1.1 Use table 2 below for sampling. If a production lot is composed of more than 5 containers of raw beef trimmings, randomly select 5 containers from the lot for sampling.

Table 2: Number of sample pieces to collect per container
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

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

5.1.3 Using a sanitized knife, cut off a thin slice that is at least 20 cm² for each of the 60 pieces [for example at least 8 cm (3") × 2.5 cm (1") × 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.

5.1.4 Place the sampled slices into the sterile sampling bag. Sampling in the above manner produces a sample of 60 pieces of trim weighing 325 g to 1 kg.

Note: Operators must check with the contracted laboratory to find the sample weight required.

5.2 N5 method of sample collection

5.2.1 For beef products not amenable to excision sampling (e.g., finely textured beef, meat from Advance Meat Recovery (AMR), Partially Defatted Chopped Beef (PDCB), ground beef), a minimum of five sample units of approximately 200 g each must be collected from the lot. Sample units must be representative of the whole lot.

5.2.2 Aseptically collect the sample.

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

6. Lot definition

A lot is defined as for the sampling under M218 for all products except for ground beef where lot is defined as for the sampling under M201.

7. Target organisms

The samples will be analysed for the presence of 6 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 The method of analysis 5C.04 for the detection, isolation and identification of STEC published in the USDA FSIS Microbiology Laboratory Guidebook (MLG) can be used.

Other methods of analysis may be accepted by FSIS. An establishment may select from the methods list provided in Foodborne Pathogen Test Kits Validated by Independent Organizations (PDF), under the STEC tab. However, to be considered acceptable, the method must be:

Validated for testing relevant foods by:
1. a recognized independent body (that is, Association of Official Agricultural Chemists (AOAC), Association Française de Normalisation(AFNOR), MicroVal, NordVal), or
2. a U.S. regulatory body (that is FSIS MLG or Food and Drug Administration Bacteriological Analytical Manual (FDA BAM)), or
3. Canadian Association for Laboratory Accreditation (CALA) and the Standards Council of Canada (SCC), or
4. an International Organization for Standardization (ISO) method
The validated method should be:
1. fit for the intended purpose and application, and
2. performed under validated conditions by a laboratory that assures the quality of the analytical results

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

has on their scope of accreditation any of the applicable testing methods described above for analysing raw beef products for the STECs of interest or
has initiated the application process with the Standards Council of Canada or the Canadian Association for Laboratory Accreditation to add any of these methods to their scope of accreditation and the laboratory has previously been accredited for at least 1 testing method for other micro-organisms (for example, Salmonella, Listeria monocytogenes, E. coli O157, etc.) in food.

9. Reporting of the results

Operators are required to convey Annex D-2 addendum 1 (STEC in beef products verification sampling template) and Annex D-2 addendum 2 (Instructions for completion of STEC in beef products 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, for example, 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 BT-PDB@inspection.gc.ca.

Positive (present) results should be classified as High Priority or Urgent, and submitted within 1 business day. Negative (absent) results may be combined into 1 template and submitted within the last week of each month.

10. Follow-up on positive results

10.1 Beef products that tested presumptive positive (that do not confirm as negative if confirmation methods are performed), and/or confirmed positive for any of the 7 STEC of interest must not be exported to the USA and 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 products as per requirements outlined in the Preventive controls for E. coli O157/NM in raw beef products the guidance for E. coli O157:H7. Presumptive positive results that confirm culturally to be negative (for example, 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 products are on hold. In the case that the implicated products were distributed, the inspector will immediately notify the Area Operational Specialist responsible for meat export and Area Recall Coordinator.

Addendum 1

STEC in beef products verification sampling template sample information

STEC in beef trimmings verification sampling template. Description follows.

All beef establishments that are eligible to export beef to the USA and producing raw beef manufacturing trimmings, raw beef bench trimmings. Raw ground beef and raw ground beef components other than trim as the final product are required to report Shiga toxin-producing E. coli (STEC) verification laboratory test results

Addendum 2

Instructions for completion of STEC in beef products verification sampling template

Color coding of the template

Peach cells: data needs to be entered by the lab,
Yellow cells: drop-down menus provide the data options to be entered by the lab,
Blue cells: the data auto populates and is dependent on information entered in other cells,

Data entry

Laboratory information: enter the laboratory name and city information at the top of the template using the following format: laboratory name, city
Completion and verification of sample information:
1. enter unique job/project number as found on record/certificate of analysis
  This number is issued by the lab for internal tracking purposes
2. verify the lab name, city (column C) and enter the province (column D)
3. select the appropriate registered establishment number, of the meat establishment for whom the samples have been analyzed, from the drop-down menu
  The establishment name and province will auto populate based on the registered establishment number entered
4. verify the establishment name and province
  If this information is not correct, re-check the registered establishment number that was entered. If the establishment you are searching is not available please contact CIFA at: BT-PDB@inspection.gc.ca
5. enter the unique sample ID as assigned by the laboratory for tracking purposes
6. enter the product name and a brief description of the product which was analyzed
7. select the appropriate temperature range from the drop-down menu which encompasses the temperature of the product upon reception at the lab
8. enter the following dates: the date sampled, date received at lab, and date analysis was completed
9. from the drop-down menu select the screening method used
10. from the drop-down menu, identify the analytical result of the screening method
  If a presumptive positive is being reported as a confirmed positive, and no confirmatory method is being performed, go directly to step 2.m.
11. if the screening method was positive, use the drop-down menu to identify the confirmatory method used
12. identify the analytical result of the confirmatory method from the drop-down menu
13. identify which E. coli serotype was identified, using the drop-down menu
  If more than 1 serotype is identified separate lines of data entry are required for each positive. If a presumptive positive is being reported as a confirmed positive, select "presumptive positive" from the menu

Please note that if, for whatever reason, the laboratory is unable to analyse the sample submitted for analysis, the operator is expected to submit a replacement sample as soon as possible.

Reporting

3. All results must be reported to the Operator as soon as they are available
4. Results entered into the template along with the supporting record/certificate of analysis must be reported to the Food Safety Division, CFIA through the dedicated email account: BT-PDB@inspection.gc.ca
5. The subject line of the e-mail should be of the format: Lab name – Job number
6. Emails reporting positive (present) results should be classified as high priority or urgent, and submitted within 1 business day.
7. Negative (absent) results may be combined into 1 template and submitted within the last week of each month
8. Please do not report confirmation of sample receipt through this email account