1. Purpose
   
   This instruction provides import inspection personnel with the procedures for sampling beef and veal products when assigned an E. coli O157:H7 type-of-inspection (TOI) by the Automated Import Information System (AIIS).
    
2. Cancellation
   
   Part 3, Section 5, "Sampling Program for Escherichia coli O157:H7 in Imported Raw Ground Beef or Veal Products; Imported Raw Ground Beef or Veal Components; and Imported Beef or Veal Patty Components (Project MT-08/MT-51)," dated June 08, 2006
    

3. Forms
   
   FSIS Form 10,210-3, "Requested Sample Programs" (see FSIS Directive 10,210.1) for both sampling projects (see Table 8)
   
   Project Number MT08 is used to submit samples of imported raw ground beef or veal
   
   Project Number MT51 is used when testing imported raw beef or veal trimmings or components (fresh or frozen) or raw ground beef or veal patty components
   
   After the form is completed, inspection personnel must make a photocopy of the form for their case file prior to shipping the sample to the laboratory. Instructions for completion of the form are in Enclosure 2. The production date(s) of the samples selected for analysis must be documented on the lab form in block 28.

4. Imported Raw Products that are Subject to E. Coli 0157:H7

   1. Project MT08 sampling of imported raw ground beef and veal products sampled for E. coli O157:H7. These products are covered under Process Category 03B and include products such as ground beef/veal, hamburger, beef patties, and beef patty mix [319.15 (a), (b), and (c) products].

   2. Project MT51 sampling of imported raw ground beef/veal components and raw ground beef/veal patty components that are used as the raw material source in the production of raw ground product. These products are covered under Process Category 03C. Examples of products defined as raw ground beef/veal components include esophagus meat, head meat, cheek meat, beef/veal for manufacturing or trimmings, boneless beef/veal, beef from Advanced Meat Recovery systems (AMR), low temperature rendered lean finely textured beef (LFTB), partially defatted chopped beef, partially defatted beef fatty tissue, and heart meat.
       

5. Supplies

   1. FSIS Form 10,210-3, "Requested Sample Programs."

   2. Ordering MT08 & MT51 Forms

      FSIS Form 10,210-3 for MT08 and MT51, when needed, are ordered by inspection personnel and are distributed directly to the import establishment from the Office of the Chief Information Officer (OCIO). In the event that the supply of forms at an import establishment has been depleted and inspection personnel need them immediately, they should contact the Office of International Affairs (OIA), Import Inspection Division (IID) for assistance. DO NOT photocopy blank laboratory forms to submit with samples as each form has a unique tracking number in the bar code at the upper right corner and cannot be reused. Only original forms will be accepted by the FSIS laboratories unless otherwise specified by IID Headquarters.

      When ordering these forms, inspection personnel will send their request to the import inspection mailbox, ImportInspection@fsis.usda.gov, with a carbon copy (cc :) to their applicable Regional Import Field Office (RIFO) mailbox. Such requests shall be made by the end of each month. The e-mail will include the I-house number, type of form requested (MT08 or MT51), and the requested amount. Orders should be placed in increments of 25 for either MT08 or MT51, as applicable.

      Inspection personnel will allow sufficient time for delivery and monitor form usage. It is recommended that a three (3) month supply of the applicable lab forms be maintained at the I-House.

      All other sampling supplies needed (gloves, bags, shippers/boxes, seals, air bills, aseptic sampling supplies, etc.) are furnished by the designated FSL to inspection personnel. If there is a need for more sampling supplies, inspection personnel should contact the FSL identified in "block 9" of FSIS Form 10,210-3 by sending an Outlook message to the appropriate lab (i.e., "Sampling Supplies - Eastern Lab"). Supplies for sampling should be requested in advance based on the types of inspections an import establishment can conduct. Inspection personnel should refer to FSIS Directive 7355.1 for any additional guidance.
       

6. Sampling Procedures
    
   1. Obtaining Assignments
      
      Microbial assignments for E. coli O157:H7 are generated by the AIIS. Inspection personnel must check the increased and intensified list of countries and establishments daily to verify any special sampling requirements for MT08 or MT51. Inspection personnel will refer to Part 3, Section 1, "Laboratory Sampling General Procedures," for the different levels of sampling of imported meat and poultry products.
       

   2. Sampling Programs

      1. MT08 Sampling Program

         1. Sanitation Guidelines

            It is extremely important that good hygiene and product handling practices are exercised at the import establishment to avoid contaminating sample units even when conducting intact sampling. Inspection personnel will refer to Enclosure 1, aseptic handling procedures, as a guideline to handling products during sample collection. Affected equipment must be properly cleaned and sanitized before and after sample collection to prevent cross-contamination of the sample and inspected lots after collection of the sample.

         2. Sample Size

            1. The preferred sample size is approximately a one (1) pound intact sample unit from the assigned lot. However, if the:

               1. i. intact packages are less than one (1) pound, select enough packages to acquire an approximate one (1) pound sample.

               2. ii. Immediate container is greater than one (1) pound and the importer/exporter wishes an entire intact package to be selected as the sample (and it is practical to ship the sample unit to the FSL), select an intact sample unit.

               3. iii. Immediate container is of a size that is not practical to ship to the FSL as an intact unit, select the sample using the aseptic methods described in Enclosure 1. If aseptic sampling supplies are not available, refer to FSIS Directive 7355.1 for acquiring aseptic sampling supplies.

         3. Selecting Samples

            1. Samples will be selected using aseptic sampling procedures.

            2. E. coli O157:H7 samples will be collected when assigned by the AIIS.

         4. Preparing Samples

            1. Fresh (Unfrozen) Products

               1. Do not freeze samples of fresh product which arrived Monday through Friday as fresh unfrozen product. Samples must be collected and submitted to the laboratory Monday through Friday while remaining in the fresh unfrozen state.

               2. If a shipment arrives on Saturday, Sunday, or holidays, or after the contract carrier has picked up packages at the establishment for the day, samples of product arriving in a fresh unfrozen state must then be frozen and shipped to the laboratory on the next work day.

            2. Frozen Products

               1. Do not allow samples to thaw.

               2. Samples of product must be kept frozen and shipped to the laboratory on the first workday after the weekend or holiday.

         Inspection personnel shall proceed to Part VI., Submitting Samples for instruction on submitting samples.

      2. MT-51 Sampling Program
         
         The following instructions address sampling for beef manufacturing trimmings and other ground beef or beef patty components.

         NOTE: These instructions replace the "Procedures for Sampling Raw Ground Beef Components and Raw Beef Patty Components" in Attachment 2 of FSIS Directive 10,010.1, Revision 1.

         The sampling collection procedure outlined below is the same as the method shown in the Trim and Sub-primal Sample Collection Training CD that FSIS issued to inspection personnel at beef manufacturing trimmings suppliers participating in the nationwide raw ground beef component microbiological baseline. Inspection program personnel who previously completed this training are not required to retake the training. Inspection personnel need to:

         • e-mail the Continuing Education and Distance Learning mailbox (CEDL@fsis.usda.gov) in Outlook to request the Trim Sample Collection Training CD; and

         • complete the training as soon as practical.

         NOTE: The Agency will allocate up to one hour of official time (code 01 time) to inspection personnel to complete the requisite training.

         1. Sanitation Guidelines

            It is extremely important that good hygiene and product handling practices are exercised at the import establishment to avoid contaminating sample units even when conducting intact sampling. Inspection personnel will refer to Enclosure 1, aseptic handling procedures, as a guideline to handling products during sample collection. Affected equipment must be properly cleaned and sanitized before and after sample collection to prevent cross-contamination of the sample and inspected lots after collection of the sample.

            Implements/equipment may be sanitized by immersing the clean implement(s)/equipment into a hot water bath that maintains a temperature of  > 180o F (82o C) for the full minute. This treatment will not sterilize the implement/equipment, but it will inactivate the most heat resistant non-spore forming bacteria that FSIS tests for, such as Escherichia coli O157:H7, Listeria monocytogenes and Salmonella.

            If a hot water bath is not available or is inappropriate for the equipment type, the establishment may use a sanitizer-disinfectant according to the manufacturer's directions for pre-cleaned equipment. One acceptable sanitizer is chlorine bleach diluted with 9 parts tap water, with a contact time of 30 seconds. After sanitizing the implements/equipment, excess sanitizer must be removed from sample contact surfaces with potable water to avoid sanitizer contamination of the sample. Chlorine solution used for sanitizing implements/equipment used for this procedure must be from a freshly mixed solution to assure proper strength.

         2. Sampling

            1. Inspection personnel are to collect samples from one specific production code/date.

            2. When collecting samples, inspection personnel are to:

               • " assure that equipment used for removal of the sample(s) have been properly cleaned and sanitized according to the import establishment's Sanitation Standard Operations Procedures (SSOP); and

               • " use gloves and handle all sanitized surfaces so that they do not become contaminated.

         3. Beef Manufacturing Trimmings - Fresh (not frozen). To sample beef manufacturing trimmings, select samples using the N60 method of sample collection (as described below) and collect 60 individual pieces from raw beef manufacturing trimmings;

            1. if a specific production lot is composed of more than 5 containers of raw ground beef components, randomly select 5 containers with the same production code/date for sampling;

            2. if the specific production lot is composed of 5 or less containers, use the chart below for sampling:

            Number of Sample Pieces to Collect Per Container
            # of containers in each specific production	# of sample pieces to select from each container
            5	12 pieces
            4	15 pieces
            3	20 pieces
            2	30 pieces
            1	60 pieces

            3. aseptically collect the appropriate number of pieces of beef manufacturing trimmings based on the number of containers that represent one specific production code/date. Use the sanitized hook or tong to lift a piece of meat off the top of the container. The total number of pieces collected is 60 and each piece should weigh approximately 15 grams. During slivering, pay attention to take as much surface area for each 15 grams as possible;

            4. cut off a slice that is approximately a 4 inch length by 2 inch width and 1/8 inch thick from each of the 60 pieces. Cut off as much of the beef manufacturing trimmings' outer surface as possible. The priority is to collect samples from pieces of product taken from the original surface of the beef carcass. Samples are to be very thin slices (approximately 1/8 inch thick);

            5.  collect and bag the sample slices. Weigh the sample to ensure approximately 2 pounds of product are collected in the whirlpak sample bag; and

            6. check the product temperature of the top pieces of meat from randomly selected containers of beef components (do not take the temperature of the actual sample slices). Record the temperature on the sample request form in block 21. If the sample pieces came from more than one container, record the temperature of the warmest container in block 21 of the sample request form. If the product is warmer than 40o F, secure the bag containing the sample and place the sample in a cooler to chill before shipping;

         4. Beef Manufacturing Trimmings - Frozen. To sample frozen beef manufacturing trimmings, select the appropriate number of containers from the lot. The containers must have the same production code/date. Collect 60 individual pieces from frozen beef manufacturing trimmings as follows:

            1. Remove from the container or expose the top surface of the randomly selected frozen block(s) of beef trimmings.

            2. For each individual block, aseptically remove about 15 grams of slivered product at each of 12 evenly distributed locations around the periphery of the frozen block. The diagram in Enclosure 3 illustrates how an N12 sample would be taken at every 30° point around the entire circumference of the frozen block. Repeat N12 sampling for each of the additional blocks until sixty 15-gram samples are taken. During slivering, pay attention to take as much surface area for each 15 grams as possible. The method of removal of the N60 samples will be at the discretion of the import establishment (i.e., band saw, chisel, electric saw, knife) as well as the preparation1 (i.e., defrosting, tempering) of the containers, when applicable.

            3. Under circumstances when five blocks cannot be sampled, remove 60 slivered samples from one or more blocks in a similar fashion. For less than five blocks, remove the block(s) from the container and sample the bottom or one or more side faces of the block to better ensure that the 60 samples represent different sites on the product.

            4. 4. Composite the contents of each block into a single sample bag

         5. Advance Meat Recovery (AMR) Product and Low Temperature Rendered Beef (LTR) Products. To sample AMR or LTR products, randomly select a sample consisting of 2 pounds of product from a specific production code/date.

         6. Other Raw Beef Components, including primal and sub-primal cuts, and carcasses. To sample other raw beef components, randomly collect one piece, or enough pieces, of the beef components to equal 2 pounds. If the component is very large, cut slices in the manner described in c. or d. above.

         ¹ Air Tempering (partially thawing) frozen blocks of beef must occur in a refrigerated area (< 40^o F).

7. Submitting Samples
   
   1. Inspection personnel must complete one FSIS Form 10,210-3 for each lot sampled. Ensure that the product temperature in block 21 and the requested information in Block 28 of the sample request form are complete.
       

      1. Complete instructions and illustrations for securing samples and attaching seals are contained in FSIS Directive 7355.1, Revision 2, "Use of Sample Seals for Laboratory Samples and Other Applications".

      2. Inspection personnel will refer to Part 1, Section 1, "Laboratory General Procedures," for related procedures concerning preparing, securing, and submitting samples.

      3. FSIS Form 10,210-3 must be enclosed in a separate bag and inserted in a self-sealing bag along with the sample unit. Only original forms will be accepted. Please be sure to maintain a copy of the form for the establishment case file.

      4. Pack the sample with ice packs.

   2. Laboratory

      Sample unit(s) must be mailed to the FSL identified on the FSIS Form 10,210-3.

   3. Mailing

      Samples will be mailed via the FSIS contract overnight delivery/courier mailing service. Samples must be shipped Monday through Friday so that they arrive at the laboratory overnight. A sample box shipped on Friday must have a "Saturday Delivery" sticker on it and "Saturday Delivery" marked on the shipping label to avoid delays and discarded samples. Samples must not be shipped on Saturday or on the day before a Federal holiday. To avoid discard of samples upon receipt at the laboratory, inspection personnel must document the reason for any delays in shipping the sample to the laboratory in Block 28, "Remarks," of the lab form.
       

8. Laboratory Results Reporting Procedures
   
   Laboratory results are automatically forwarded to the AIIS by the Laboratory Electronic Application for Results Notification (LEARN) system. Inspection personnel will provide LEARN results to establishment management even if the establishment receives e-mail notifications. The Biological Information Transfer and email System (BITES) messages will report positive test results to the RIFO.

   Inspection personnel will check LEARN in accordance with FSIS Directive 10,200.1 to obtain test results.
    

   1. Presumptive Positive Notification
      
      When a sample is reported as presumptive positive, IID headquarters and/or the RIFO are notified through the Biological Information Transfer and E-mail System (BITES) and the result is posted in LEARN. IID and/or the RIFO will notify inspection personnel who in turn will notify establishment management of the reported presumptive positive sample.
      
   2. Negative Results (Acceptable)
      
      When a sample tests negative for all analyses, the involved lot is not subject to further testing and may be released if on Voluntary or FSIS Hold and all other reinspection criteria are acceptable. The RIFO will notify establishment management of the results through inspection personnel.
       
   3. Positive Results (Not Acceptable)
       

      1. When a sample is reported as positive, IID headquarters and/or the RIFO are notified through the Biological Information Transfer and E-mail System (BITES) and the result is posted in LEARN. IID and/or the RIFO will notify inspection personnel who in turn will notify establishment management of the reported positive sample. 

      2. If a Normal or Increased sample tests positive for E. coli O157:H7, the lot is:

         • Refused entry, if shipment/lot was on FSIS or Voluntary Hold

         • Subject to Recall, if shipment/lot was not held (RIFO and IID Headquarters must use IID Recall Procedures)

         • The foreign establishment that produced the lot is placed on "Intensified" until 15 consecutive lots of product from the same process category from that establishment are sampled for E. coli O157:H7 and reported as Acceptable.

      3. When an Intensified sample tests positive for E. coli O157:H7, the lot is refused entry. The foreign establishment that produced the product will continue on Intensified until 15 consecutive lots of product from the same process category from that establishment are sampled and reported as Acceptable for E .coli O157:H7.

      4. IID headquarters will notify import inspection personnel of any action to take relating to the "positive" result for the shipment (notifying plant management, faxing any remaining paperwork, etc).

Enclosure 1

Aseptic Sampling Guidelines

1. Samples taken for microbiological analyses require stringent sampling techniques known as aseptic sampling procedures. Import inspection personnel must refer to the following aseptic guidelines when collecting all, including intact microbial samples.

   1. For each sample:

      • Wash and scrub hands thoroughly to the mid-forearm, using antibacterial hand soap (or a hand sanitizer at 50 ppm chlorine equivalency, if available).

      • Open the "whirl-pack" bag without contaminating the interior. 

      • Peel open the package of sterile gloves from the top without contaminating the exterior of the gloves. 

      • Remove a glove by holding it from the wrist side opening inner surface. Avoid any contact with the outer surface of the glove.

      • Insert hand without puncturing the glove.

      • Do not touch anything with the exterior of the glove except the sample.

      • If you have concern that the glove may be contaminated, discard that glove and use another sterile glove.

      • If non-intact, sanitize the knife, hook or tongs before collecting the samples.

      • With the gloved hand, collect the sample. Place the sample into the opened bag. 

      • Remove and discard the glove and close the bag.

Enclosure 2

Instructions for completing FSIS Form 10,210-3, Requested Sample Programs, for Project Number MT08 and MT51

Block Numbers

1- 2. Completed by OPHS. 

3. If not already completed, enter import establishment number (e.g. 00038 I). 

4. Leave blank. 

5. If not completed by OPHS, enter 92. 

6. If not completed by OPHS, enter two letter state abbreviation code. 

7. If not completed by OPHS, enter two number regional field office code (02 or 06- Detroit; 03 or 07 - Los Angeles; 04 or 08 - Miami; 05 or 09 - Philadelphia) 

8. If not completed, enter information as requested. 

9. Completed by OPHS. 

10 - 12. Leave blank. 

13. Completed by OPHS 

14. Completed by OPHS. 

15. Enter the name of the country of origin on the product. 

16. Enter the two character UN country code from Part 3, Section 4, Enclosure 3, for the country of origin. 

17. Enter the foreign establishment number of the plant that produced the product. 

18. Completed by OPHS. 

19. Enter the date the sample was collected from the lot. 

20. Enter the date the sample was mailed to the lab. 

21. Leave blank. 

22. Check the appropriate block. NOTE: Lots assigned at Intensified must be placed on FSIS Hold. 

23. Enter the health certificate number. 

24. Enter the appropriate lot number (e.g. 1, 2) from FSIS Form 9540-1. 

25. Check the appropriate box. Note: Hold is checked for an E. coli O157:H7 sample assigned at the Intensified level. 

26 - 27. Leave blank. Do not complete. 

28. Complete all blanks as requested. Do not leave out required information. Production dates, bar codes, and other information are very important for recall purposes. The production code/date of the product selected from the lot for analysis must be documented in this block. 

29 - 32. Complete as requested. 

33. Contact your supervisor before submitting any forms with this block completed. 

34 - 41. Laboratory use only.

Enclosure 3

Frozen Beef Trimmings Sampling - n12

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