Microbial Sampling for

2. Regulatory Requirements HACCP Plan Changes Determine if Plan is Controlling Hazards or to Audit Supplies Regulatory Requirements Generic E. coli and Salmonella Testing in Slaughter Environments Not Necessarily a Part of HACCP Plan Listeria Testing Must be a Part of HACCP/sSOP Plan to Avoid FSIS Product Testing HACCP Plan Changes Must be Statistically Sound Determine if Plan is Controlling Hazards or to Audit Supplies Periodic Sampling Regulatory Requirements Generic E. coli and Salmonella Testing in Slaughter Environments Not Necessarily a Part of HACCP Plan Listeria Testing Must be a Part of HACCP/sSOP Plan to Avoid FSIS Product Testing HACCP Plan Changes Must be Statistically Sound Determine if Plan is Controlling Hazards or to Audit Supplies Periodic Sampling

3. What is a sampling plan?? Sampling Plan Should be a Written Document What to Include… “…statement of criteria of acceptance applied to a lot based on appropriate examination of a required number of sample units by specified methods.” Sampling Plan Should be a Written Document Number of Samples Drawn from Lot Number of Samples Permitted to Exceed Limit (If Applicable) The Limit Stringency of Plan is Adjusted by Altering these Factors ICMSF “…statement of criteria of acceptance applied to a lot based on appropriate examination of a required number of sample units by specified methods.” Sampling Plan Should be a Written Document Number of Samples Drawn from Lot Number of Samples Permitted to Exceed Limit (If Applicable) The Limit Stringency of Plan is Adjusted by Altering these Factors ICMSF

4. Sponge/Surface Samples Liquid Samples Solid Samples Sample Collection Sample Handling Sponge/Surface Samples Food Products Carcasses Food Contact Surfaces Liquid Samples Solid Samples Excision Samples Ground Samples Representative of Lot Must Be Handled Correctly Must Consider Homogeneity and Uniformity of Lot Should be Collected Aseptically Sterile Spoon, Forceps, Sponge, Scissors, etc. Sterile Containers Properly Identify Deliver Sample to Lab Promptly/Refrigerate Transport in Ice Chest Maintain at 0-4 C Do NOT Freeze Analyze within 36 h Sponge/Surface Samples Food Products Carcasses Food Contact Surfaces Liquid Samples Solid Samples Excision Samples Ground Samples Representative of Lot Must Be Handled Correctly Must Consider Homogeneity and Uniformity of Lot Should be Collected Aseptically Sterile Spoon, Forceps, Sponge, Scissors, etc. Sterile Containers Properly Identify Deliver Sample to Lab Promptly/Refrigerate Transport in Ice Chest Maintain at 0-4 C Do NOT Freeze Analyze within 36 h

5. 100% Assurance is NOT Possible Low Occurrence E. coli Example Data Interpretation Microorganisms are NOT Randomly Distributed in Products 1.2 um3 - space occupied In 1cc of well mixed liquid, 106 bacteria would only occupy 0.0001% of the sample Occur as Individual Cells or Clumps Report as CFU/ml Microorganisms are NOT Randomly Distributed in Products 1.2 um3 - space occupied In 1cc of well mixed liquid, 106 bacteria would only occupy 0.0001% of the sample Occur as Individual Cells or Clumps Report as CFU/ml E. coli O157:H7-Low Probability Event 1 Positive in 1000 Does NOT necessarily indicate that the lot is more contaminated than Negative Lots Positive could have been a matter of probability Single Samples Do not Give Meaningful Information Sampling is Useful to Establish Baselines Screen Raw Materials Verify Controls NO SAMPLING PLAN CAN ENSURE SAFETY OF FINAL PRODUCT Microorganisms are NOT Randomly Distributed in Products 1.2 um3 - space occupied In 1cc of well mixed liquid, 106 bacteria would only occupy 0.0001% of the sample Occur as Individual Cells or Clumps Report as CFU/ml Microorganisms are NOT Randomly Distributed in Products 1.2 um3 - space occupied In 1cc of well mixed liquid, 106 bacteria would only occupy 0.0001% of the sample Occur as Individual Cells or Clumps Report as CFU/ml E. coli O157:H7-Low Probability Event 1 Positive in 1000 Does NOT necessarily indicate that the lot is more contaminated than Negative Lots Positive could have been a matter of probability Single Samples Do not Give Meaningful Information Sampling is Useful to Establish Baselines Screen Raw Materials Verify Controls NO SAMPLING PLAN CAN ENSURE SAFETY OF FINAL PRODUCT

6. NEGATIVE TESTS DO NOT GUARANTEE AN ABSENCE OF PATHOGEN, ONLY A CERTAIN AMOUNT OF CONFIDENCE THAT’S WHY WE USE HACCP PLANS!!!!

7. For HACCP Verification and Validation, test for INDICATOR ORGANISMS Sample Collection Usually Described in Regulations or in USDA Guidebook or BAM Sampling Frequency Changing a CCP For HACCP Verification and Validation, test for INDICATOR ORGANISMS Aerobic Plate Counts Generic E. coli/Listeria Coliforms Can Supplement with Pathogen Testing - Salmonella Carcasses and Subprimal Cuts - Use USDA Sponge Sample Methods Ground Beef - 60 g Most Sizes Defined in USDA Laboratory Guidebook Info Available from Laboratory Conducting Test Extension Specialists Sampling for HACCP Plan Verification No Set Standards Companies Conduct Final Product Testing Weekly, Monthly or on Each Batch Same Intervals may be Used to Test Suppliers 1-5 Samples/Sampling Period Individual or Composite Samples May be Used Must Consider Labor and Cost Issues NO requirement for Basic Microbial Testing in HACCP Plan To Change/Validate a CCP or CL Must be Statistically Sound Data must be Collected on 3 Separate Days of Processing Generally, based on current Literature, 28 samples/day is considered to be statistically sound If Standard Deviation is High then more samples should be collected For HACCP Verification and Validation, test for INDICATOR ORGANISMS Aerobic Plate Counts Generic E. coli/Listeria Coliforms Can Supplement with Pathogen Testing - Salmonella Carcasses and Subprimal Cuts - Use USDA Sponge Sample Methods Ground Beef - 60 g Most Sizes Defined in USDA Laboratory Guidebook Info Available from Laboratory Conducting Test Extension Specialists Sampling for HACCP Plan Verification No Set Standards Companies Conduct Final Product Testing Weekly, Monthly or on Each Batch Same Intervals may be Used to Test Suppliers 1-5 Samples/Sampling Period Individual or Composite Samples May be Used Must Consider Labor and Cost Issues NO requirement for Basic Microbial Testing in HACCP Plan To Change/Validate a CCP or CL Must be Statistically Sound Data must be Collected on 3 Separate Days of Processing Generally, based on current Literature, 28 samples/day is considered to be statistically sound If Standard Deviation is High then more samples should be collected

8. Must Have Limits Determines Acceptable or Unacceptable Limits are Microbiological Criteria Guidelines Standards Purchase Specifications Standards Limits set by regulatory agencies May lead to recall if exceeded Guidelines Internal limits set by company Failure alerts processor Purchase Specifications Agreement between vendor and buyer Regulatory Limits 0 Tolerance Generic E. coli and Salmonella Presence of LM in RTE Products Guidelines Use the USDA Baseline Data as a Guide for Raw Products Cooked Products should be Pathogen Free Standards Limits set by regulatory agencies May lead to recall if exceeded Guidelines Internal limits set by company Failure alerts processor Purchase Specifications Agreement between vendor and buyer Regulatory Limits 0 Tolerance Generic E. coli and Salmonella Presence of LM in RTE Products Guidelines Use the USDA Baseline Data as a Guide for Raw Products Cooked Products should be Pathogen Free

9. Randomness Can Be Achieved a Number of Ways: Pull Number out of a “hat” Computer Generated Numbers Random Number Tables Get a Random Number, Collect data from that specific package/area. Areas must be assigned a number

10. The Guidelines in this Plan Can Be Applied to Control of Other Pathogens

11. Ready to Eat Products-Product Intended to be consumed without any further safety preparation steps Process Categories-USDA Not Heat Treated - Shelf Stable Heat Treated - Shelf Stable Fully Cooked - Not Shelf Stable Product with Secondary Inhibitors - Not Shelf Stable (only RTE in this category)

12. Sample - “a collection of product that represents a …sampled lot…that has passed the establishments pre-shipment review. Must be in consumer-ready package and processed in the same manner as the rest of the lot Sampled Lot - “…all product produced under a single HACCP plan between performance (…clean-up to clean-up)”

13. HACCP Systems Verified Collecting and Testing RTE products for microbial hazards May test of Salmonella and/or E. coli O157:H7 from sample Positive Sample Corrective actions and Preventive measures must be taken Recall of product represented by sample

14. Establishment Tests AT LEAST one RTE product per HACCP plan for pathogen every month. OR Establishment tests one RTE product per HACCP plan once every 3 months AND Conducts on-going food contact surface and non-food contact surface testing for indicator organisms and Conducts targeted product testing for LM when a positive result for Listeria spp. occurs

15. Establishment MUST have a written protocol for testing that includes Frequency of testing Randomness of sampling Recording of results Actions taken if positive Establishments must develop scientifically-based frequency for testing

16. Scientific Basis Journal Articles Recommendation from Scientific Experts Scientific Journal Articles FSIS Guidance Materials Establishment History

17. Product Testing must be incorporated into the HACCP plan as an on-going verification activity Environmental and food-contact surface testing must be a part of sSOPs. It can be a part of the HACCP plan. Documentation to support testing protocol must be in the facility Preventive and corrective actions must be taken when a positive result occurs

31. Must Be Written and Include Elements Previously Mentioned Must be a Part of HACCP and/or sSOPs in order to avoid FSIS product Testing Must be Scientifically Sound