The EURLEuropean Union Reference Laboratory manual provides information on relevant EUEuropese unie legislation, analytical methods, standardisation of methods in ISOInternational Standardisation Organisation and CENEuropean Committee for Standardisation and reference materials.

EU legislation

Relevant EUEuropese unie legislation related to the EURLEuropean Union Reference Laboratory -Salmonella and NRLNational Reference Laboratorie -Salmonella activities can be found on the website: http://europa.eu. Here, documents are available in several languages. Some of the important legislation (in English) is listed below, while several recent regulations can be found at: http://ec.europa.eu/food/food/biosafety/hygienelegislation

  • COUNCIL DIRECTIVE 92/117/EEC  (pdf, 40Kb) of 17 December 1992 on measures for protection against specified zoonoses and specified zoonotic agents in animals and products of animal origin to prevent outbreaks of food-borne infections and intoxications
  • DIRECTIVE 2003/99/EC  (pdf, 123Kb) OF THE EUROPEAN PARLIAMENT AND OF THE COUNCIL of 17 November 2003 on the monitoring of zoonoses and zoonotic agents, amending Council Decision 90/424/EEC and repealing Council Directive 92/117/EEC
  • REGULATION (EC) No 2160/2003  (pdf, 156Kb) OF THE EUROPEAN PARLIAMENT AND OF THE COUNCIL of 17 November 2003 on the control of Salmonella and other specified food-borne zoonotic agents
  • REGULATION (EC) No 882/2004  (pdf, 338Kb) OF THE EUROPEAN PARLIAMENT AND OF THE COUNCIL of 29 April 2004 on official controls performed to ensure the verification of compliance with feed and food law, and with animal health and animal welfare rules
  • REGULATION (EC) No 2073/2005  (pdf, 450Kb) - Commission Regulation of 15 November 2005 on microbiological criteria for foodstuffs 
  • COMMISSION REGULATION (EU) no 208/2011  (pdf, 836Kb) of 2 March 2011 amending Annex VII to Regulation (EC) no 882/2004 of the European Parliament and of the Council and Commission Regulations (EC) no 180/2008 and (EC) no 737/2008 as regards lists and names of EU reference laboratories            

Reference materials

Definitions of reference materials (RM) and certified reference materials (CRM) according to ISOInternational Standardisation Organisation Guide 30.

Reference material (RM)

A material or substance of wich one or more properties is/are sufficiently well established to be used for the calibration of apparatus, the assessment of a measurement method or to assignment of values to materials.

Certified reference material (CRM)

A reference material, one or more of wich the property values are certified by a technically valid procedure and accompanied by, or traceable to, a certificate or other documentation issued by a certifying body.

From 1986 up to 1995, the Microbiological Laboratory for Health Protection (MGB) of the Dutch National Institute for Public Health and the Environment (RIVMRijksinstituut voor Volksgezondheid en Milieu) led EUEuropese unie funded projects on the development and certification of microbiological reference materials. The experience and the knowledge obtained from these projects have been applied in preparing reference materials for use as standard samples in the interlaboratory comparison studies of the EURLEuropean Union Reference Laboratory -Salmonella.
The reference materials are prepared as follows:

  • A selected strain is cultured and mixed with milk
  • This milk suspension is spray-dried, resulting in a highly contaminated milk powder (hcmp). Which is then stored at -20 ºC
  • A small portion of the hcmp is mixed with sterile milk powder until the desired contamination level
  • The mixed powder is put into gelatin capsules, resulting in the final reference materials (to be stored at -20 ºC)

The first certification studies on microbiological reference materials were organised in 1992, resulting in the successful certification of 6 batches of reference materials:

  • CRM506 Enterococcus faecium
  • CRM507 Salmonella Typhiumurium
  • CRM527 Enterobacter cloacae
  • CRM528 Bacillus cereus
  • CRM594 Escherichia coli
  • CRM595 Listeria monocytogenes

These CRMs are available at the Institute for Reference Materials and Methods (IRMM) of the Joint Research Centre (JRC) in Geel, Belgium.

After their development, the non-certified reference materials were produced for some years by the Foundation affiliated to the RIVM. This production stopped in 2003. Today several other institutes or (commercial) manufacturers also produce and sell microbiological reference materials, e.g. Institute Pasteur in France and Health Protection Agency in the UK.

http://www.irmm.jrc.be/

http://www.hpacultures.org.uk/products/lenticulediscs/index.jsp

ISO/CEN

The EURLEuropean Union Reference Laboratory -Salmonella is an active member (and project leader in some areas) of ISOInternational Standardisation Organisation /TC34Technical Committee 34 on Feed and Food products /SC9Subcommittee 9 on microbiology and CENEuropean Committee for Standardisation /TC275Technical Committee 275 on food analysis - horizontal methods /WG6Working Group 6 on microbial contaminants . See below for the explanation of these acronyms.

ISO : International Standardisation Organisation

TC34 : Technical Committee 34 on Feed and Food products

SC9 : Subcommittee 9 on microbiology

CEN : European Committee for Standardisation

TC275 : Technical Committee 275 on food analysis - horizontal methods

WG6 : Working Group 6 on microbial contaminants

The committees mentioned in ISO and CEN not only standardise methods for the detection and enumeration of micro-organisms, but also for items like validation of methods, quality assurance of media and guidance for the organisation of proficiency tests.       

Analytical methods

See below for information on EUEuropese unie and/or international accepted methods for:

Detection of Salmonella

The international accepted method for the detection of Salmonella in food and animal feed is described in EN-ISOInternational Standardisation Organisation 6579:2002, Microbiology of food and animal feeding stuffs - Horizontal method for the detection of Salmonella spp. An amendment to this EN-ISO was published in 2007 in an annex, in which the detection of Salmonella spp. in animal faeces and in environmental samples of the primary production is described (Annex D of ISO 6579). 

Since 2008, EN-ISO 6579, including annex D, is under revision. Detection of Salmonella will become part 1 of the new CENEuropean Committee for Standardisation -ISO 6579 series.

CEN ISO 6579-1 Microbiology of the food chain -- Horizontal method for the detection, enumeration and serotyping of Salmonella -- Part 1: Horizontal method for the detection of Salmonella spp. is expected to be published before Summer 2016.

Enumeration of Salmonella

Enumeration of Salmonella spp. is described in CEN ISO/TS 6579-2:2012 Microbiology of food and animal feed -- Horizontal method for the detection, enumeration and serotyping of Salmonella -- Part 2: Enumeration by a miniaturized most probable number technique. The procedure describes a miniaturized MPN method in 12-wells microtiter plates with MSRV as selective enrichment medium.

Serotyping of Salmonella 

The reference method for serotyping of Salmonella uses the ‘White-Kauffmann-Le Minor scheme’, as most recently described by Grimont and Weill (2007). The scheme can be found at: https://www.pasteur.fr/sites/default/files/veng_0.pdf

Guidance on serotyping of Salmonella spp. is described in CEN ISO/TR 6579-3:2014 Microbiology of the food chain -- Horizontal method for the detection, enumeration and serotyping of Salmonella -- Part 3: Guidelines for serotyping of Salmonella spp.

Year

Author

Title

2007, 9th edition

Grimont, P.A.D. and Weill, F-X
 

Antigenic formulae of the Salmonella serovars
 

2001, 8th edition

Popoff MY

Antigenic Formulas of the Salmonella Serovars

1997

Popoff MY and Le Minor L

Guidlines for the Preparation of Salmonella Antisera

Phagetyping of Salmonella 

The scheme for phagetyping Salmonella (especially Salmonella Typhimurium and Salmonella Enteritidis) accepted at EU level is the one used by the WHOWorld Health Organization reference centre situated at the Public Health England  (PHEPublic Health England , formerly Health Protection Agency) in London (Colindale), UK.

http://www.phe.org.uk/

Sampling whole melons to check for the presence of Salmonella

On 9 October 2012 a request was received from EC DG-Sanco to give advice on the best way to sample whole melons to check for Salmonella. A document was drafted after consulting experts and literature and sent to DG-Sanco on 22 October 2012. Recently the document was updated to include sampling of watermelons as well. This link shows the updated document (pdf, 97KB).