Contact person: eurl-listeria@anses.fr This document is the fourth version of the European Union Reference Laboratory for Listeria monocytogenes (EURL Lm) Technical Guidance Document (TGD) on challenge tests and durability studies for assessing shelf-life of ready-to-eat foods for Listeria monocytogenes. It replaces the third version of 6 June 2014 – Amendment 1 of 21 February 2019. The first version of this TGD (2008) was prepared at the request of the Directorate General Health & Consumers (DG SANCO) of the European Commission (EC) in response to the needs expressed by EU Member States to have a document providing both detailed and practical information on how to conduct shelf-life studies on Listeria monocytogenes (Lm) in ready-to-eat (RTE) foods to ensure compliance to the microbiological criteria set out in Article 3.2 of Regulation (EC) No 2073/2005. The purpose of this revision is to ensure consistency between the EURL Lm Technical Guidance Document and the standard EN ISO 20976-1 on "Requirements and guidelines for conducting challenge tests of food and feed products - Part 1: challenge tests to study the growth potential, lag time and maximum growth rate" published in 2019. The standard specifies the protocols for carrying out challenge tests for growth studies for any bacteria and yeasts that do not form mycelium, whereas the EURL Lm Technical Guidance Document covers the technical aspects specific to Lm in RTE foods, which are not addressed in the standard. Therefore, the EURL Lm Technical Guidance Document should now be read in conjunction with the standard and be considered as a supplementary document to the standard EN ISO 20976-1. The revision of this TGD also includes the experience gained over the years in carrying out challenge tests. This document remains complementary to the EC/DG SANCO document, entitled “Guidance document on Listeria monocytogenes shelf-life studies for ready-to-eat foods, under Regulation (EC) No. 2073/2005 of 15 November 2005 on microbiological criteria for foodstuffs”. The fourth version of the TGD was prepared by EURL Lm together with a working group of six NRLs Lm and was approved by the Standing Committee on Plants, Animals, Food and Feed on 1 July 2021.

Funding by: EUROPEAN HEALTH AND DIGITAL EXECUTIVE AGENCY (HADEA) - HaDEA.A – Health and Food Directorate A2.002 - EU4Health/SMP Food Unit Project: [EURLs-EURCs/2021-2022/SI2.870826] Document dispatch by DG SANTE G4 via https://food.ec.europa.eu/document/download/77d7e9a5-9db3-49c2-89c4-780f10a48071_en?filename=biosafety_fh_mc_tech-guide-doc_listeria-in-rte-foods_en_0.pdf