EURL-CAMPYLOBACTER National Veterinary Institute

EURL- Campylobacter Hanna Skarin

CAMPYLOBACTER

CAMPYLOBACTER SPECIES

• At present, 27 recognized Campylobacter species (including 14 subspecies), some have been moved to other genera: Arcobacter and Helicobacter

• Thermotolerant Campylobacter species: microorganism forming characteristic colonies on solid selective media when incubated in a microaerobic atmosphere at 41,5 °C: C. jejuni and C. coli (C. lari, C. upsaliensis and others)

• Other Campylobacter species are not (or much less)

significant as zoonotic pathogens

THE EUROPEAN UNION SUMMARY REPORT ON TRENDS AND SOURCES OF ZOONOSES, ZOONOTIC AGENTS AND FOOD‐BORNE OUTBREAKS IN 2016

EFSA (European Food Safety Authority) and ECDC (European Centre for Disease Prevention and Control), 2017. The European Union summary report on trends and sources of zoonoses, zoonotic agents and food‐borne outbreaks in 2016. EFSA Journal 2017;15(12):5077, 228 pp.

The most frequently reported

foodborne pathogen in the EU

(>240.000 cases)

FOOD-BORNE OUTBREAKS IN EU 2010-2016

EFSA (European Food Safety Authority) and ECDC (European Centre for Disease Prevention and Control), 2017. The European Union summary report on trends and sources of zoonoses, zoonotic agents and food‐borne outbreaks in 2016. EFSA Journal 2017;15(12):5077, 228 pp.

EURL-CAMPYLOBACTER EURL established in 2006 according to

Regulation (EC) No. 882/2004

THE PRESENT EURL-CAMPYLOBACTER-TEAM

Hanna Skarin Gunilla Lindgren Joakim Skarin Susanna Westerberg Therese Jernberg Linda Svensson Elina Lahti Helena Höök Sevinc Ferrari Bo Segerman

EURL-CAMPYLOBACTER THE NETWORK

• The network includes 35 NRLs from MSs • + Iceland, Norway, Switzerland, Serbia, FYROM, Bosnia and Herzegovina, Albania

LEGAL BASIS

• Regulation (EC) 2017/625 on official controls includes the tasks and duties of EURLs

• The Zoonosis Directive 2003/99/EC on the monitoring of zoonoses and zoonotic agents (Campylobacter is a priority agent that should be monitored)

MICROBIOLOGICAL CRITERIA • Commission Regulation (EU) 2017/1495 amending Regulation (EC) No

2073/2005 as regards Campylobacter in broiler carcases

• Public health risk reduction from the consumption of broiler meat of more than

50 % could be achieved if carcases complied with a limit of 1 000 cfu/g in chicken neck skin*

* EFSA Scientific opinion on control options for Campylobacter along the poultry meat production chain (2011)

other sources

20-30%

50 - 80% EFSA Scientific Opinion on the risk of human campylobacteriosis linked to broiler meat (2010)

KEY ACTIVITIES FROM THE WORK PROGRAM

• Organise proficiency tests (PTs) • Validate (and improve or produce) analytical

methods • Maintain reference collections of

Campylobacter strains • Provide scientific and technical assistance to

NRLs • Provide scientific and technical assistance to

the European commission and other organisations

ORGANISATION OF PROFICIENCY TESTS

To check the potential and capacity to perform Campylobacter analyses on relevant samples

To collect information from NRLs, to identify problems

ORGANISATION OF PROFICIENCY TESTS ISO 10272: 2017 Microbiology of the food chain — Horizontal method for detection and enumeration of Campylobacter spp. • PT Detection and identification (ISO 10272 part 1) Matrix: Primary production • PT Enumeration and detection (ISO 10272 part 2) Matrix: Food

MATRICES FOR THE PT Food Primary production Chicken breast meat Sock samples Chicken minced meat Caecum samples Chicken skin Environmental swab samples Milk filter Feacal swab samples Minced beef

MATRICES FOR THE PT • The Swedish Campylobacter monitoring programme for broiler

chicken • All broiler flocks are tested for Campylobacter • Analysis are performed at SVA • Matrices are chosen from producers that are free from

Campylobacter

Analysed flocks Campylobacter-positive flocks

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ORGANISATION OF PROFICIENCY TESTS- PREPARATIONS • Stability and homogeneity of the

freeze-dried bacteria • Stability of tests containing live

cultures – up to 5 days • Final test: all strains and all vials

with the same matrix as in the PT (same background flora) are tested

The Pack Day

ADDITIONAL TESTS

PTs on molecular methods (voluntary): • PCR • Next year: subtyping (MLST and/or WGS-based typing)

VALIDATE (AND IMPROVE OR PRODUCE) ANALYTICAL METHODS

• Methods for detection PCR for detection (without pre-enrichment) • Methods for species identification PCR: in silico evaluation, validation of two PCR-assays (annex to ISO 10272) MALDI-TOF MS (next slide)

C. UPSALIENSIS/HELVETICUS • Sequenced the same number of C. upsaliensis and C.

helveticus strains that got score >2 in MALDI-TOF MS and compared them to published genomes

• Only 30% of the C. helveticus strains were true C. helveticus, the rest were C. upsaliensis

• Ongoing: sequence strains in MALDI-TOF MS

database

VALIDATE (AND IMPROVE OR PRODUCE) ANALYTICAL METHODS • Methods for genotyping Multi-locus sequence typing (MLST) Methods for WGS clustering analysis WG- whole genome sequencing PulseNet International

• Campylobacter survival in different matrices Chicken skin Milk

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REFERENCE COLLECTIONS

• Well characterised strains (to be used i validation studies, for PTs, as control strains)

• Freeze-drying of Campylobacter (application in PTs) Different species and bacteria of related genera Different levels Homogeneity and stability testing based on ISO 22117,

ISO 17034 and the ISO Guide 35

• The annual workshop in Sept/Oct in Uppsala • 1,5 days (Mon evening-Wed lunch) • In connection to CHRO (Aberdeen and Nantes)

TRAINING AND SUPPORT TO NRLs WORKSHOP

TRAINING AND SUPPORT TO NRLs TRAINING COURSES • Training courses (1-2 courses per year) Upon request by NRL or initiated by the EURL • Courses in: Enumeration, detection and species identification of

Campylobacter Molecular methods: PCR, PFGE, MLST

TRAINING AND SUPPORT TO NRLS EURL WEBSITE- LEARNING MATERIALS

http://www.sva.se/en/service-and-products/eurl-campylobacter

PROVIDE SCIENTIFIC AND TECHNICAL ASSISTANCE TO THE EUROPEAN COMMISSION AND OTHER ORGANISATIONS

• Communication on ad hoc basis • EFSA Scientific Network for Zoonoses Monitoring Data • Outbreak investigations • CEN/ISO standardization activities

Thank you!