Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli Workshop: Monitoring and Reporting.

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Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli Workshop: Monitoring and Reporting of food-borne outbreaks – Ankara, 22 Feb EU Reference Laboratory for Escherichia coli Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare Istituto Superiore di Sanità, Roma Laboratory-based surveillance of foodborne outbreaks in Europe Alfredo Caprioli

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli Surveillance and control of foodborne diseases in the EU Medical sector Food/Veterinary sector The reference laboratory network for foodborne pathogens in the medical field The reference laboratory network for foodborne pathogens in the veterinary/food field The activity of the EU-RL for E.coli Content

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli Surveillance and control of foodborne diseases in the EU DG - SANCO Food and Veterinariy Sector Medical sector Programme on Food - Warterborne Diseases and Zoonoses Member States Zoonoses Directive (99/2003) Monitoring of Zoonoses Zoonoses Task Force

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli 20 Diseases

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli Six priority diseases Salmonellosis Campylobacteriosis E.coli STEC/VTEC infection Listeriosis Shigellosis Yersiniosis FWD surveillance network Disease specific epidemiologists and microbiology experts designated by Member States

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli International surveillance network for human infections with Salmonella, VTEC, and Campylobacter. Foodborne and Waterborne Diseases Network Since 2007

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli General FWD surveillance objectives Enhance –detection of international food-borne clusters and outbreaks –exchange of information on causative agents/strains Strengthen –integrated surveillance in humans, food and animals –laboratory capacity in MSs

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli ECDC case definitions

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli FWD surveillance network  Standard EU case definitions  Notification of cases in the TESSy database  External laboratory quality assurance  Urgent inquiries (international outbreaks)  Annual meetings

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli FWD-2009

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli Campylobacter 190,566198,252 Salmonella 131,468108,614 Yersinia 8,346 7,595 E.coli VTEC 3,159 3,573 Listeria 1,381 1,645 FWD Infections with: No.cases reported in

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli FWD-2009

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli Foodborne and Waterborne Diseases Network VTEC infections by serogroup

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli FWD-2009

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli Minimum Requirements of National Reference Laboratories Pilot proposal for : Salmonella, Listeria and VTEC

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli Requirements of National Reference Laboratories 1.Connected with public health authorities 2.Receive strains for typing from all the country, particularly from outbreaks 3.Capable to identify strains 4.Capable to maintain strain collections 5.Capable to type strains. For Salmonella: - Typing of the “top 30” serotypes - Distinguish S. Paratyphi B from S.Java - Distinguish S.Typhimurium from monophasic

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli 6.Perform antimicrobial susceptibility testing 7.Participate in external quality assurance programs 8.Perform molecular “subtyping”: PFGE MLVA, MLST Requirements of National Reference Laboratories

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli PULSENET Elettroforesi in campo pulsato (PFGE) Detect foodborne disease case clusters by PFGE Facilitate early identification of common source outbreaks

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli PULSENET INTERNATIONAL Elettroforesi in campo pulsato (PFGE)

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli Surveillance and control of foodborne diseases in the EU DG - SANCO Food and Veterinariy Sector Member States Zoonoses Directive (99/2003) Monitoring of Zoonoses Task Force Zoonosi

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli The “Zoonoses Directive” 2003/99/CE Directive 2003/99/CE of 17 november 2003 on measures for monitoring and surveillance of zoonoses and zoonotic agents Art 4.1: MS shall collect relevant and comparable data in order to identify and characterise hazards… Art 4.2: Monitoring shall take place at the stages of the food chain most appropriate to the zoonosis concerned

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli The “Zoonoses Directive” 2003/99/CE Annex I A, First rank priority Salmonella Escherichia coli VTEC Campylobacter Brucella Listeria monocytogenes Mycobacterium bovis Echinococco Trichinella Monitoring an surveillance of zoonotic agents in animals and food

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli Zoonoses Report 2009 Salmonella in food

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli Zoonoses Report 2009 Salmonella serotypes in pigs

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli Zoonoses Report 2009 Salmonella serotypes in pig meat

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli Zoonoses Report 2009 Campylobacter species in animals

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli EC legislation in food safety To ensure that food produced in or imported into the EU have high and comparable safety standards in all Member states Legislation covering the whole food chain – Directive 2003/99/EC – Regulation (EC) No 2160/2003 – Regulation (EC) 852/2004 – Regulation (EC) 853/2004 – Regulation (EC) 854/2004 – Regulation (EC) 2073/2005 – Regulation (EC) 882/2004

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli Community Reference Laboratories (EU-RL) for specific food and feed hazards and for specific animal diseases (Art. 32). National Reference Laboratories (NRL), which are designated by Member States for each CRL (Art. 33) Regulation (EC) 882/2004 on official controls TITOLO III REFERENCE LABORATORIES

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli Functions and duties of EU-RLs (882/2004, Art. 32) To provide NRLs with analytical reference methods To provide NRLs with reference materials To organise comparative testing (ring tests) To conduct training for NRLs, (annual workshop, courses, visits to CRL) To provide scientific and technical assistance to the Commission

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli Functions and duties of NRLs (882/2004, Art. 33) To collaborate with their EU-RL To coordinate the activities of laboratories involved in official food controls in their own country To organise comparative tests for those labs To disseminate the information received from EU-RLs To provide scientific support to their Authorities

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli EU EU-RL Member States (27) NRLs Laboratories involved in controls of food and feed or animal diseases Local level The Network of EU Reference Laboratories Accreditation ISO 17025

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli 41 EU-RLs, selected and designated for 5-years periods upon public calls Regulation EC No 776/ EU-RLs for animal health 14 EU-RLs for chemical risks in food and feed 13 EU-RLs for biological risks in food and feed

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli 1. Classical swine fever 2. Foot and mouth disease 3. African horse sickness 4. Avian influenza 5. Newcastle disease 6. Swine vesicular disease 7. Fish disease 8. Bivalve mollusc diseases 9. Bluetongue 10. African swine fever 11. Zootechnics 12. Equine diseases 13. Rabies 14. Brucellosis EU-RLs for Animal Health

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli EU-RLs for for chemical risks in food and feed 1. Residues of veterinary medicines and contaminants in food of animal origin 2. Residues listed in Annex I, Group A 1, 2, 3, 4, Group B 2(and Group B 3 to Directive 96/23/EC 3. Residues listed in Annex I, Group B 1 and B 3(e to Directive 96/23/EC and carbadox and olaquindox 4. Residues listed in Annex I, Group A 5 and Group B 2(a), (b), (e) to Directive 96/23/EC 5. Residues listed in Annex I, Group B 3(c) to Directive 96/23/EC 6. Additives for use in animal nutrition 7. Material intended to come into contact with foodstuffs 8. Residues of pesticides in cereals and feedingstuffs 9. Residues of pesticides in food of animal origin 10. Residues of pesticides in fruits and vegetables 11. Residues of pesticides, single residue methods 12. Heavy metals in feed and food 13. Polycyclic Aromatic Hydrocarbons (PAH) 14. Dioxins and PCBs in feed and food

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli 1.Salmonella 2.Campylobacter 3.Listeria 4.Escherichia coli (VTEC) 5.Staphylococci 6.Foodborne Parasites 7.Antimicrobial resistance in bacteria of animal origin 8.Bacterial and viral contamination of molluscs 9.TSE 10.Marine biotoxins 11.Animal proteins in feeds 12.Genetically modified organisms (OGM) 13.Milk EU-RLs for for biological risks in food and feed

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli EU-RL for Escherichia coli (VTEC) Istituto Superiore di Sanità, Rome, Italy

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli EU-RL for Salmonella RIVM, Bilthoven Netherland

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli EU-RL for Campylobacter National Veterinary Institute Uppsala, Sweden

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli EU-RL for Listeria monocytogenes AFSSA, Maisons-Alfort, France

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli EU-RL for coagulase-positive Staphylococci AFSSA, Maisons-Alfort, France

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli Functions and duties of EU-RLs (882/2004, Art. 32) To provide NRLs with analytical reference methods To produce and distribute reference materials To organise comparative testing (Proficiency Tests) To conduct training for NRLs, (annual workshop, courses, visits to CRL) To provide scientific and technical assistance to the Commission

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli Proficiency testing and training for NRLs Inter-laboratory studies on VTEC identification and typing Identification of VTEC by PCR detection of VT and intimin coding genes

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli No. of Laboratories vtx1vtx2eae Proficiency tests Virulence genes Proficiency testing and training for NRLs PCR detection of VTEC virulence genes

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli O157O26O145O121O111O103O91 No. of Laboratories Proficiency tests Serogroups Proficiency testing and training for NRLs Identification of VTEC serogroups

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli Detection of pathogenic non-O157 VTEC in food

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli ISO/DTS for VTEC in food Recommended by EFSA for the detection of VTEC in food and animal samples

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli 2011: Vegetables (spinach) 2008: Bacterial strains Proficiency tests organized by the EU-RL VTEC 2009: Carcass swabs 2010: Milk Detection of pathogenic VTEC in food

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli Proficiency testing and training for NRLs Real Time PCR detection of VTEC in food No. of NRLs Interlaboratory study

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli STEC O104:H4 outbreak, Germany 2011 Source: Robert Koch Institute. Report: Final presentation and evaluation of epidemiological findings in the EHEC O104:H4 outbreak, Germany Berlin Urgent inquiry on 24 May EWRS on 22 May

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli What happened with VTEC O104:H4 Real Time PCR for vtx genes Real Time PCR for eae gene Attaching/Effacing adhesion VTEC O104:H4 Real Time PCR Specific for O104 and H4 genes In close collaboration with the medical Reference Laboratory at the SSI in Copenhagen

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli EU-RL involvment in the outbreak May 24 Report of the outbreak in ProMed May 25 Alert to NRLs May 27 1 st Draft method for VTEC O104 sent to NRLs June 1 Outbreak strain received from Robert Koch Inst. June 2 Method evaluated, revised and sent to NRLs June 3-6 Reference DNA from VTEC O104 sent to 14 NRLs as positive control June 9-15 Reference DNA sent to other 28 Labs Methods

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli Laboratory Preparedness ! Importance of VTEC O1O4 outbreak: what can we learn ? For facing food safety crisis

Istituto Superiore di Sanità - Dip. Sanità Pubblica Veterinaria e Sicurezza Alimentare EU Reference Laboratory for E.coli Thank you for attention !