Experiences and potential of food microbiology laboratories

Illustration with EURLs/NRL networks for Listeria monocytogenes, Staphylococci, Milk & Milk Products

NRL Conference, Backweston, 3-4 February 2016

Bertrand LOMBARD Manager EURLs L. monocytogenes, Staphylococci, Milk & Milk Products

ANSES-Laboratory for Food Safety, Maisons-Alfort (FR) bertrand.lombard@anses.fr

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Outline

• Activities of EURLs Listeria monocytogenes,

Staphylococci, Milk & Milk Products

• Strengths & weaknesses

of the EURL/NRL networks

• Perspectives for EURL/NRL networks

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ACTIVITIES OF EURLs

LISTERIA MONOCYTOGENES,

STAPHYLOCOCCI,

MILK & MILK PRODUCTS

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• Mandates

• 3 types of activities

1. Coordination of NRL networks

2. Analytical development

3. Scientific & technical support to DG SANTE

& other EC organizations

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ANSES-Laboratory for Food Safety

• 2 locations o Maisons-Alfort

o Boulogne-sur-Mer

• Accredited EN ISO 17025 by COFRAC o For most of its activities

o In particular regulatory analyses involved in EURL mandates

• Quality system for organization of proficiency testing (PT) trials

• Designated o Community Reference Laboratory (CRL)

o European Union Reference Laboratory (EURL)

o For 3 mandates

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Laboratory for Food Safety, Maisons-Alfort and Boulogne-sur-Mer sites

Laurent LALOUX, Director , Anne BRISABOIS, Deputy director

QUALITY Team Cécile Cecconi, Quality Manager, Head

LISTERIA, SALMONELLA, E. COLI Unit Renaud Lailler, Head David Albert, Deputy

ADMINISTRATIVE AND FINANCIAL Service Christelle Gautheron, Head

METALLIC AND MINERAL TRACE ELEMENTS Unit Laurent Noêl, Head

STAPHYLOCOCCUS, BACILLUS, CLOSTIDIA, MILK Unit Jacques-Antoine Hennekinne, Head Frédéric Auvray, Deputy

QUANTITATIVE RISK ANALYSIS AND MODELLING Team Annie Beaufort, Head

ANIMAL HOUSE Service Sébastien Allix, Head Océane Le Bidel, Deputy

PESTICIDES E& MARINE BIOTOXINS Unit Frédéric Hommet, Head Chanthadary Phandanouvong, Dputy

ENTERIC VIRUSES Unit Sylvie Perelle, Head

CENTRAL VETERINARY SERVICES LABORATORY Unit Agnès Chamoin, Head

HIGH-THROUGHPUT PLATFORM FOR IDENTIFICATION AND TYPING OF PATHOGENS Patrick Fach, Head

TECHNICAL SUPPORT TO LABORATRY ACTIVITIES Service Sylvie Rudelle, Head

SECURITY AND SAFETY BIOLOGICAL Team Sébastien Allix, Head

HYGIENE , SAFETY AND OCCUPATIONAL HEALTH Laurence Bouhier, Christine Delpont, Ghislaine Merle, health and safety assistants

2 services and 2 common teams with the Maisons-Alfort Laboratory for Animal Health

Updated: November 2014

Unit for the BACTERIOLOGY AND PARASITOLOGY OF FISHERY PRODUCTS AND AQUACULTURE

Graziella Bourdin, Head, Mélanie Gay, Deputy

Unit for the PHYSICAL CHEMISTRY OF FISHERY PRODUCTS AND AQUACULTURE Guillaume Duflos, Head

HYGIENE , SAFETY AND OCCUPATIONAL HEALTH Graziella Bourdin, health and safety assistant

QUALITY Régine Quenu, Quality Manager

ADMINISTRATIVE AND FINANCIAL Service Sophie Holuigue, Head

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Department of Chemical Food Contaminants Thierry Guérin, Head

Department of Microbiological Food Contaminants Michel-Yves Mistou, Head

ANTIMICROBIAL RESISTANCE Team Sophie Granier, Head

SUPPORT AND DEVELOPMENT OF RESEARCH ACTIVITIES Team Sylviane Dragacci, Head

COORDINATION OF REFERENCE ACTIVITIES Team Bertrand Lombard, Head

Unit for the BIOCHEMISTRY OF AQUATIC PRODUCTS Thierry Grard, Head

Department of Fishery products and Aquaculture Anne Brisabois, Head Guillaume Duflos, Deputy

Mandates (2)

• For milk & milk products (MMP) – 1992 to 2005 (CRL)

• Directive 92/46 on hygiene of milk chain

– Bacteria, staphylococcal enterotoxins, phosphatase

– From 2006 (EURL by Reg. 776/2006 modifying Reg. 882/2004)

• Regulatory frame – EC Regulation 853/2004: Annex II/Section IX on raw milk & MP

» Hygiene rules for food of animal origin, FBOs

– EC Regulation 854/2004: Article 8 & Annex IV on raw milk and MP

» Official controls for food of animal origin

» National Competent Authorities to monitor own checks of raw milk producers

• Criteria for mesophilic total flora (TF), somatic cells (SC) indicators of raw milk’s hygiene at collection stage and use of raw cow’s milk for processing

• Phosphatase activity tracer of correct pasteurization of raw cow’s or buffalo’s milk and heat treatment of raw milk & MP

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Mandates (3) • For Listeria monocytogenes (Lm)

& coagulase positive staphylococci (CPS) (incl. staphylococcal enterotoxins -SEs) – Designated in 2006 (Reg. 776/2006)

– Reg. frame: EC Regulation 2073/2005 on microbiological criteria for food

– Lm: 3 food safety criteria applicable to ready-to-eat (RTE) food • RTE for infants & special medical purpose

• RTE able/unable to support Lm growth

• 2 types of limits – Qualitative: absence/25 g

– Quantitative: 100 cfu/g at the end of shelf-life

– CPS: 5 process hygiene criteria, quantitative limits for • Cheeses

• Milk/whey powder

• Cooked crustaceans & molluscan shellfish

– SEs: 1 food safety criterion, absence/25 g • Cheeses, milk/whey powder

• When CPS > 105 cfu/g

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1. Coordination of NRL networks (1)

• 3 networks

– At least 1 NRL from each EU Member State

– Lm: 35 NRLs

• Also Norway

– CPS: 36 NRLs

• Also Norway

– MMP: 36 NRLs

• Also Norway, Switzerland

– For Ireland

• DAFM, Backweston

• Bernadette Hickey

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1. Coordination of NRL networks (2)

• Annual workshops

– General scope or thematic

– Regulatory/epidemiological context

– Works undertaken since last workshop

– Work programme for next year

• Proficiency testing (PT) trials

– 1/year for regulatory parameters

– Use of reference methods required

+ Lm PFGE typing, PCR for CPS se genes

– Investigation of new test material • Matrices

• Contamination techniques

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1. Coordination of NRL networks (3)

• Training sessions

– Upon needs expressed by NRLs & for NRLs with

unsatisfactory results to PT trials

– Examples

• Lm PFGE typing + data analysis

• Shelf-life studies for Lm

• Detection of SEs/se genes

• SC counting

• Video tutorials, on EURL websites

– Lm PFGE typing & profile interpretation

– SE detection

– SC counting

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1. Coordination of NRL networks (4)

• Scientific & technical advice to NRLs

– Confirmatory analyses • Lm PFGE typing

• CPS typing, se genes detection, SE quantification/typing

• In particular for investigating national outbreaks

• Recent example in Ireland – Staphylococcal food-borne outbreak, flight IE to US, end

2015

– Strawberry crumble mousse

– At EURL: PFGE typing, PCR se genes, SE quantification/type (SEA)

– Support for organization of national PT trials

– Replies to queries (e-mail, phone)

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1. Coordination of NRL networks (5)

• Participation to national meetings

• EURL Lm database on PFGE profiles of

Lm food isolates

– Voluntary participation of NRLs (12)

• Including IE-NRL: largest provider of

profiles after France

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1. Coordination of NRL networks (5)

• EURL guides for NRLs/national official labs

– Measurement uncertainty/Lm enumeration

– Sampling techniques of surfaces of food

processing environment for Lm analysis

– Shelf-life studies: challenge tests, durability study

– Lab competence to conduct shelf-life studies

• For CAs & NRLs

• Developed in collaboration with some NRLs

(WGs)

– Active participation of Bernadette Hickey

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2. Analytical development (1)

• EURL in-house work, possibly in

collaboration with NRLs

• To develop/improve & validate reference

methods/reference materials for official

control

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2. Analytical development (2)

• Examples: methods

– SE detection

• Lack of Standard method EURL method, cited in

Reg. 2073/2005

– Fluorimetric methods for AP in milk, cheese & cream

+ AP limits for correct pasteurization

EC Reg. 2074/2005

• Cream: IE request

– Characterization/typing methods

• Lm: PFGE, fALFP, MLVA, WGS

• CPS: PFGE, spa, MLVA, PCR (se genes) MALDI-ToF, WGS

– Confirmatory methods

• SE quantification, SE types: in-house ELISA, MS

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2. Analytical development (3)

• Certified reference materials

in collaboration with EC/JRC/IRMM (Geel)

– SE in cheese: almost finalized

– SC in milk

– DNA-plugs for Lm PFGE typing

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3. Scientific & technical assistance to DG SANTE (1)

• Upon the needs

• Development/implementation of European legislation – Ex 1: preparation of Reg. 2073/2005

– Ex 2: Implementation of Annex II of Reg. 2073/2005 • EURL Lm Guidance document on challenge tests, durability

studies

• Classification of RTE food in Lm criteria

• Participation for DG SANTE to European/international technical meetings/activities – CEN/ISO/IDF standardization

• Involvement in EC/third countries sanitary negotiations

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3. Scientific & technical assistance to DG SANTE (2)

• Participation to EFSA WGs

• Curation of EFSA/ECDC database for

PFGE profiles of Lm food strains

• Investigation of European-level food-borne

outbreaks (Lm)

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STRENGTHS & WEAKNESSES

OF THE EURL/NRL NETWORKS

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Strengths (1)

• 2-level network

– One lab financed by EC at European level

– Transferring methods/training/assessing a NRL network

– Each NRL itself coordinating a network of official labs

• Confidence (CAs/third countries importing European products) in official controls in Europe

– Consistent & comparable system

– Assessed use of reference methods

– EURLs/NRLs: accredited EN ISO 17025

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Strengths (2) • Close relations between EURL team/NRLs and

between NRLs – Several opportunities to meet/year

– NRL representatives are stable

– Excellent quality of human relationships!

• Sharing experiences, knowledge

• Harmonization of practices/methods used

• Enquiries to all NRLs on specific topics – State of the art at European level

• 2-level expertise – Questions official labs/CAs NRLs

– NRLs EURL

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Strengths (3)

• Drafting reference documents

– Not EURL alone, but with volunteering NRLs, and

approval by all NRLs

– European scale, national aspects taken into account

• Practical solutions to problems of NRLs

– Practical training (EURL/on-site)

– Video tutorials

– Tel/email

• EURL work programmes

– Based on NRL needs, expressed in particular at

workshops

– NRL feedback on activities, in particular at workshops

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Weaknesses • EURLs per danger/food categories-species:

reduced sharing of tools/means

• Uneven level of participation of NRLs to EURL activities – Workshops

– PT trials • Lack of participation

• Lack of cooperation for follow-up of deviating results

– Lack of response to EURL enquiries

• Differences between NRLs – Size, means, competence

– For designing PT trials, training

• > 1 NRL/country – Coordination at national level

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PERSPECTIVES

FOR EURL/NRL NETWORKS

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• To take advantage of the quality of EURL/NRL network to build R&D research projects

– COST: WGS for Lm

– H 2020/European Joint Programme: WGS/MS/MALDI-ToF for CPS

• New rules for EURL/NRLs in the revision of Reg. 882/2004

• New EURLs envisaged

– Viruses/all food

– Plant health

– Human health

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