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  1. Laboratory Methods (Food)

BAM Chapter 5: Salmonella

Bacteriological Analytical Manual (BAM) Main Page

Authors

Wallace H. Andrews (ret.), Hua Wang, Andrew Jacobson (ret.), Beilei Ge, Guodong Zhang, and Thomas Hammack (ret.)

Editions 

Current Edition

Previous Editions

Revision History 

  • October 2026: Corrected the date of the previous Revision History bullet to be January 2026 (there was no May 2025 edition) and expanded upon the revisions originally listed for that edition. Revised Appendix A title in the Table of Contents from "Validated food matrices" to "Validated Food Matrices by FDA Loop-Mediated Isothermal Amplification (LAMP) and Real-Time Polymerase Chain Reaction (qPCR) Methods" and added additional validated matrices for qPCR in Appendix A; Deleted mushroom from Section C7a and relocation to Section C7d; Revised Section C7d title from Fruits and vegetables to Fruits and vegetables, including mushroom, not otherwise specified in Section C; Added collagen hydrolysate to Section C.14 and created a new Section C.30 for whole onion; Correction of typo "25°C" to "2-5°C" in Section C21; Inserted correct hyperlinks for LAMP and qPCR screening methods in Section D and F; Updated the LAMP protocol in Section C.2 to include DNA template preparation from agar plates; Updated the qPCR protocol to include an optional wash step in Section E.1 and PrepManTM Ultra reagent in Section E.2 for manual DNA extraction; Revised section F8h(2) from "discard cultures not confirmed as Salmonella by one of a, b, c, d, e, f or g methods" to "discard cultures not confirmed as Salmonella by a and b, or by one of c, d, e, f, or g methods"; Deleted “send to reference typing laboratory for definitive serotyping and identification” from Section F8(h)3; Removed shipping address from section F9 and F10; Deleted the second paragraph in Section F 10 and references 4 and 5. 
  • January 2026: Editorial revisions to sections A2, A9, and E1. Removed brand name for serological polyvalent flagellar (H) test in sections F4b and F4c. Renumbered original sections F9, F10, and F11 to F8, F9, and F10, respectively. Removed original section F9g; renumbered original sections F9h and F9i to F8g and F8h, respectively. Removed subsection designation "g" from original section F9i (1) (current F8h (1)). Revised original section F9i (2) (current F8h (2)) from "discard cultures not confirmed as Salmonella by a, and one of b, c, d or h methods" to "discard cultures not confirmed as Salmonella by one of a, b, c, d, e, f or g methods." Inserted reference 21 under section References. Section C.7, C.21, C.22, and C.26 updated to add same guidance for thawing method as C1; Section C.10 added flakes; Section C.24 revised preenrichment volume for swabs; Section D.1 updated LAMP protocol to include DNA extraction from presumptive Salmonella isolates (Section C) and baseline adjustment recommendations (Section F.3); Section D.2 updated qPCR protocol for the scope of manual DNA extraction methods application and revised the step 2 for quality check when preparing dehydrated master mix; and Appendix A added sealed concreated sampled with sponges as a new environmental matrix validated for LAMP. 
  • May 2024: Added a Salmonella screening section (Section D) to include generic LAMP and qPCR protocols; added a table (Appendix A) to list validated matrices by the two molecular methods; revamped Introduction to include a flow chart; updated Sections C.7 and C.24 on matrices and preenrichment broths including an alternative formula for modified buffered peptone water (M192c); deleted discontinued test kits in Section F.9. Original Section D and Section E from all previous versions were shifted to Section E and Section F, respectively because of the addition of Salmonella screening Section D.
  • September 2023: Section D.7 updated by requiring the use of both typical and atypical Salmonella culture controls.
  • April 2023: Chapter 5 was converted to PDF and labeled as BAM Chapter 5: Salmonella (April 2023 Edition). Section C.28 updated to remove standing at room temperature and pH adjustment, which are steps not performed at the time of SLV and MLV.
  • November 2022: Sections C.28 and E.9 updated to include screenshots for AB 7500 Fast setup and representative Genie instrument outputs, recommended GspSSD2.0 as the preferred LAMP master mix, and updated template preparation method to include a final centrifugation step. Updated section C.7a to include oysters.
  • March 2022: Section C.28 updated to include dairy feed as a validated animal food matrix, AB 7500 Fast as an alternative LAMP assay platform, two additional reagent choices for LAMP, and removed duplicate testing recommendation for LAMP screening of Salmonella in animal food protocol; Section E.9.d updated to include AB 7500 Fast as an alternative LAMP assay platform, two additional reagent choices for LAMP, and removed duplicate testing recommendation for LAMP confirmation of Salmonella isolates protocol; Section E.11 was updated by linking SeqSero to Protocols.IO.
  • October 2021: Section C.2.c updated to correct a typo; Section E.11 serotyping submission information and shipping point of contact updated.
  • September 2021: Section D.7 updated to include serological-formulas of ATCC control cultures 12325 and 29934; Section E.9.c updated to remove SmartCycler instructions from the Real-time PCR confirmation test.
  • June 2021: Section A.27 updated to include ultraviolet light source; Section C.7 updated by changing nut/nut meat preparation procedure from blending to soaking; Section C.23 updated by removing erroneous requirement to adjust the pH of preenrichment before incubation; Section C29 added method for spent sprout irrigation water from alfalfa, mung bean, and broccoli varieties; Section D.7 updated by adding Green Fluorescent Protein control strains.
  • May 2021: Section C.28 updated to include an alternative positive control, a video demonstration, and software updates for LAMP screening for Salmonella in animal food protocol; Section E.9.d updated to include a video demonstration and software updates for LAMP confirmation of Salmonella isolates protocol; Section C.7.a updated to include mushrooms.
  • February 2021: Removed text from section E.9.b.
  • December 2020:Minor changes in sections D.10.  TSI interpretation, Section E.9.b. use biochemical confirmation and typos in section E.3.a, E.3.b, E.8 and Reference 2.
  • July 2020: Section c.10. Added sample set up for non-powder forms of allspice, cinnamon, cloves and oregano. Added reference 17.
  • February 2020: Section C28. Added LAMP screening method for animal food.
  • February 2020: Section E9. Added LAMP confirmation of Salmonella isolates protocol.
  • December 2019: Section C7. Limited the foods covered by Section C7 to those not listed elsewhere in Section C.
  • November 2019: Section C.10.c Added detailed procedure to prepare sample composites for oregano, cinnamon, allspice and cloves.
  • November 2019 - Section E9 updated to include additional options for identification of Salmonella.
  • July 2018 - Section C7 revised to include vegetables not included in C23 or C27.
  • March 2018: Added real-time quantitative PCR for confirmation of Salmonella isolates protocol and validated preenrichment changes for leafy produce, herbs and sprouts; Vegetables removed from revised Section C7; Validated Preenrichment broth change in section C23.
  • August 2016: Added the Salmonella Flipbook,  a pictorial general guide to aid analysts in the detection and identification of Salmonella growing on the plating media and screening tubes used in the BAM Chapter 5 Salmonella method.   See Section E. (Prepared By: Matthew J. Forstner, Laboratory Services, Minnesota Department of Agriculture). (PDF, 13Mb)
  • December 2015: A section for the Statens Serum Institute Procedure was added to Section E: Identification of Salmonella.
  • May 2014: The VITEK 2 method of Presumptive generic identification of Salmonella was updated.
  • February 2014: Section on Detection and isolation of Salmonella from shell eggs was replaced, and validation data and additional references were added in an Appendix.
  • August 2012: Made available in PDF format versions of Chapter 5: Salmonella and Appendix 1 (archived) from 2009 which were incorporated by reference in 21 CFR Parts 16 and 118: Federal Register Final Rule (July 9, 2009, 74 FR 33030): Prevention of Salmonella Enteritidis in Shell Eggs During Production, Storage, and Transportation.  
  • November 2011: Addition to Section C: Preparation of foods for isolation of Salmonella: Leafy green vegetables and herbs.
  • February 2011: Removed link to Appendix 1: Rapid Methods for Detecting Foodborne Pathogens (now archived).
  • December 2007: Mamey pulp method added, and Section D revised.
  • June 2006: Eggs method revised for shell eggs and liquid whole eggs.
  • April 2003: Frog legs method, Lactic casein, Rennet casein, Sodium caseinate and Rabbit carcass methods revised, top ears and other dog chew toys added. Removed section A.25, Mechanical shaker.
  • October 2001: Extension of the applicability of the orange juice method in section C.19 to apple juice and apple cider.
  • December 1999, March, 2000, August 2000, and final revision on November 14, 2000. (see the Introduction for a summary of changes).

 

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