3M Food. with 3M. TB Effective Date: Number: April 2, 2013 Supersedes: New Technology Products Originating.

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1 Food Title: TM Enrichm ment Pouch with Buffered Peptone Water (ISO): Compatibility with TM Sample Handing Devices Number: TB Effective Date: April 2, 2013 Supersedes: New Technology Platform: Sample Handling TM Enrichment Pouch Products Originating Location: St. Paul, Minnesota The Enrichment Pouch with Buffered Peptone Water (BPW) (ISO) is a convenient alternative to the traditional preparation and sterilization of enrichment media. The solution conforms to the media formulation specified in the International Organization for Standardization (ISO), ISO 657 Microbiology of food and animal feeding stuffs Horizontal method for the detection of Salmonella spp. The Enrichment Pouch with BPW (ISO) consists of a stand alone pouch with 225 ml of media. The product can be used as a diluent or for the nonselective enrichment of microorganisms from food and environmental samples. Compatibility of the Enrichment Pouch with BPW (ISO) and TM Sample Handling devices was evaluated in a study. Food Center Bldg 2755W05 St. Paul, MN is a trademark of All rights reserved. TB Page 1 of 10

2 Food Study Design/Experiment: Testing was performed to determine organism recovery of inoculated Sample Handling device samples after enrichment using the Enrichment Pouch with BPW (ISO). Two studies were performed: a qualitative study using visual turbidity to assess microbial growth after enrichment and a quantitative study to enumerate microbial growth after enrichment. I. Qualitative Study: Each sample handling device was inoculated with one organism type (inoculum level of CFUs) and combined with Enrichment Pouch with BPW (ISO). The Enrichment Pouch with BPW (ISO) and sample handling device combination was gently mixed and then incubated at 37 C for 20 hours. Negative controls ( Enrichment Pouch with BPW (ISO) and sample handling device) weree processed in this same manner, except were not inoculated. The testing was performed for the following sample handling devices: TM HydratedSponge with Dey Engley Broth (DEB) TM HydratedSponge with Neutralizing Buffer (NB) TM HydratedSponge with Letheen Broth (LET) TM HydratedSponge with Buffered Peptone Water Broth (BPW) TM Quick Swab TM Dry Sponge TM Swab Sampler with Dey Engley Buffer TM EnviroSwab The organisms used for inoculation: Escherichia coli (E. coli), ATCC# 2522 Salmonella typhimurium (S.typhimurium), ATCC# Salmonella enteriditis (), ATCC# After incubation, each enrichment combination was visually observed for turbidly as compared to the negative controls. Food Center Bldg 2755W05 St. Paul, MN is a trademark of All rights reserved. TB Page 2 of 10

3 Food The following table provides the testing scheme used for the Qualitative Study. Table 1: Qualitative Study testing scheme Enrichment Pouch with BPW (ISO) with sample handling devices Sample Handling Device HydratedSponge with Letheen Broth # of samples Inoculating organism HydratedSponge with Neutralizing Buffer HydratedSponges with Buffered Peptone Water E. coli (x3 samples), (x3 samples), (x3 samples) Dry Sponge Swab Sampler with Dey Engley Buffer EnviroSwab Quick Swab 6 E. coli (x2 samples), (x2 samples), (x2 samples) Food Center Bldg 2755W05 St. Paul, MN is a trademark of All rights reserved. TB Page 3 of 10

4 Food II. Quantitative Study: Each sample handling device was inoculated with one organism type (inoculum level of CFUs) and combined with Enrichment Pouch with BPW (ISO). The Enrichment Pouch with BPW (ISO) and sample handling device combination was gently mixed and then incubated at 37 C for 22 hours. Negative controls ( Enrichment Pouch with BPW (ISO) and sample handling device) weree processed in this same manner, except were not inoculated. The testing was performed for the following sample handling devices: TM HydratedSponges with Dey Engley Broth (DEB) TM HydratedSponges with Neutralizing Buffer (NB) TM HydratedSponges with Letheen Broth (LET) The organisms used for inoculation: Escherichia coli (E. coli), ATCC# 2522 Salmonella typhimurium (S.typhimurium), ATCC# Salmonella enteriditis (), ATCC# After incubation, enriched samples were serially diluted and plated on T M Petrifilm TM Aerobic Count Plates to determine level of microbial growth. The following table provides the testing scheme used for the Quantitative Study. Food Center Bldg 2755W05 St. Paul, MN is a trademark of All rights reserved. TB Page 4 of 10

5 Food Table 2: Quantitative Study testing scheme Enrichment Pouch with BPW (ISO) enrichment with sample handling devices Sample Handling Device HydratedSponge with Letheen Broth HydratedSponge with Neutralizing Buffer # of samples Inoculating organism E. coli (x4 samples), (x4 samples), (x4 samples) HydratedSponges with Dey Engley Broth 12 Food Center Bldg 2755W05 St. Paul, MN is a trademark of All rights reserved. TB Page 5 of 10

6 Food Results: III. Qualitative Study: The following tables summarize the microbial growth results for each sample handling device after enrichment with Enrichment Pouch with BPW (ISO). Note: = Positive microbial growth = No microbial growth Table 3: Summary of microbial results of HydratedSponges after enrichment with Enrichment Pouch with BPW (ISO): Hydrated Sponge with LET Inoculating Organism E. coli S.enteriditis Negative Control 1 Sample # 2 3 Hydrated Sponge with NB Sample # Hydrated Sponge with BPW Sample # Food Center Bldg 2755W05 St. Paul, MN is a trademark of All rights reserved. TB Page 6 of 10

7 Food Table 4: Summary of microbial results of Dry Sponge after enrichment with Enrichment Pouch with BPW (ISO): Dry Sponge Inoculating Organism Sample 1 Sample 2 Sample 3 E. coli Negative Control Table 5: Summary of microbial results of Quick Swab after enrichment with Enrichment Pouch with BPW (ISO): Inoculating Organism E. coli Quick Swab Sample 1 Sample 2 Negative Control Food Center Bldg 2755W05 St. Paul, MN is a trademark of All rights reserved. TB Page 7 of 10

8 Food Table 6: Summary of microbial results of Swab Sampler with DEB after enrichment with Enrichment Pouch with BPW (ISO): Inoculating Organism Sample 1 Swab Sampler with DEB Sample 2 Sample 3 E. coli Negative Control Table 7: Summary of microbial results EnviroSwab after enrichment with Enrichment Pouch with BPW (ISO): Inoculating Organism Sample 1 EnviroSwab Sample 2 Sample 3 E. coli Negative Control Food Center Bldg 2755W05 St. Paul, MN is a trademark of All rights reserved. TB Page 8 of 10

9 Food IV. Quantitative Study: The following tables summarize the quantitative microbial growth results for each sample handling device after enrichment with Enrichment Pouch with BPW (ISO). Quantitative recovery is reported as logarithmic growth. Table 8: Summary of log recovery for HydratedSponges enriched with Enrichment Pouch with BPW (ISO): Sample Handling device Organism HydratedSponge with DEB E. coli HydratedSponge with LET E. coli HydratedSponge with NB E. coli All Samples SUMMARY OF LOG RECOVERY N 4 Mean 8.1 Std 0.03 Median 8. Min 8. 3* Max * One of the samples was not turbid after incubation indication that inoculation of the sample was inadvertently missed during laboratory testing. All negative controls were negative. Food Center Bldg 2755W05 St. Paul, MN is a trademark of All rights reserved. TB Page of 10

10 Food Conclusion: Qualitative Study: Data demonstrate that for the organism strains used in this study, all inoculated Sample Handling devices resulted in visual microbial turbidity after incubationn when enriched with the Enrichment Pouch with BPW (ISO). Quantitative Study: Data demonstrate that for the organism strains used in this study, all inoculated Sample Handling devices resulted in a minimum of 10 8 CFU/mL organism growth recovery after incubation when enriched with the Enrichment Pouch with BPW (ISO) media. Testing results demonstrate that the Enrichment Pouch with BPW (ISO) is compatible with a variety of Sample Handling devices. Food Center Bldg 2755W05 St. Paul, MN is a trademark of All rights reserved. TB Page 10 of 10