http://tiny.cc/faowgsworkshop
Potential usefulness of genome sequencing technology on food safety management - France. Presentation from the FAO expert workshop on practical applications of Whole Genome Sequencing (WGS) for food safety management - 7-8 December 2015, Rome, Italy.
Whole Genome Sequencing (WGS) for food safety management in France: Example for Listeria monocytogenes
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WGS for food safety management
in France :
Example for Listeria monocytogenes
Dr. Sophie Roussel, Anses, Maisons-
Alfort; France
Sophie.roussel@anses.fr
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Anses-presentation
12 Laboratories -18 sites
Laboratory for Food Safety
Biological agents than can affect food
safety and quality in a public health goal
Better control the food quality and
hygiene from the food industry to food
services,
Risk assessment analysis
Reference and research
Scientific and technical expertises
Monitoring and epidemio-surveillance
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WGS and Bioinformatic analysis capacities at Anses
The WGS national plateform
3 Bioinformaticians
full time
-2 technicians
-1 Ion Proton TM,
-2 High Seq
-3 Mi seq
-1 Pacbio
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WGS and Bioinformatic analysis capacities at Anses :
Laboratory for food safety
Pesticides and
marine biotoxins
Unit
Trace metals and
minerals
Physico Chemical
department
AntibioResistance
Unit
GAMeR Geno-Analysis
Modelisation : and Risk
Assessment
Identypath"
national platform
Central
Laboratory for
Veterinary
Services
Microbiology department
Enteric
viruses Unit
Staphylococci,
Bacillus,
Clostridium and
milk Unit
Salmonella-E.coli-
Listeria
(SEL) Unit
LABORATORY for FOOD SAFETY - Maisons-Alfort
-1. Statistician
-1. Bioinformatian ,
-1 expert in genotyping data
analysis
-Genotyping
-Illunina Mi seq
Listeria team
(12 persons)
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Listeria monocytogenes AnsesTeam
strains of food, feed, environment, animal
Detection-Enu meration, Ecophysiology,
Predictive Microbiology
Molecular typing Epidemiosurveillace
National Reference Lab (NRL) since 2001
French network partners
European Reference Lab (EURL) since 2006
(DG SANTE) Regulation EC 776/2006 ; https://eurl-listeria.anses.fr
Coordinate / manage European
network of 37 NRLs
Stimulate partners to do typing
Roadmap
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-
-Testing .
-Profil interpretation
-Database curation
- Data and cluster analysis
Comparison of PFGE SOPs between
European and US partners
Step 1 : Development of SOPs
(Standard Operating Procedures)
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-the French network .
-the NRL network
Proficiency testing trials
Annual training sessions;
workshops
-the National public health laboratories
(NPHL) network
-With other EURLs
-Testing .
-Profil interpretation
-Database curation
- Data and cluster analysis
Comparison of PFGE SOPs between
European and US partners
Step 1 : Development of SOPs
(Standard Operating Procedures)
Step 2 : Harmonisation
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-the French network .
-the NRL network
Proficiency testing trials
Annual training sessions;
workshops
-the National public health laboratories
(NPHL) network
-With other EURLs
-Testing .
-Profil interpretation
-Database curation
- Data and cluster analysis
Comparison of PFGE SOPs between
European and US partners
Step 3 : Molecular PFGE
databases
-European typing databases
Step 1 : Development of SOPs
(Standard Operating Procedures)
Step 2 : Harmonisation
-EFSA-ECDC database : PFGE data from
human and no human isolates
-EURL : curator of the non-human
isolates
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-the French network .
-the NRL network
Proficiency testing trials
Annual training sessions;
workshops
-the National public health laboratories
(NPHL) network
-With other EURLs
French, European and
International and PFGE-based
surveillance networks
-Testing .
-Profil interpretation
-Database curation
- Data and cluster analysis
Comparison of PFGE SOPs between
European and US partners
Step 3 : Molecular PFGE
databases
-European typing databases
Step 1 : Development of SOPs
(Standard Operating Procedures)
Step 4 : Epidémiosurveillance
improved
Duration :5 years
Step 2 : Harmonisation
-EFSA-ECDC database : PFGE data from
human and no human isolates
-EURL : curator of the non-human
isolates
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Dispatch of PFGE protocols -Communication
In 2015 : among the French / partner network : No lab is using WGS for the
surveillance in routine.
Among the European NRL network :1 lab use WGS (PHE)
21/37 NRLs carry out PFGE -Some NRLs don’t type at all
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Assess WGS technology as a typing tool
Within two different analytic development projects
Similar aims :
• 1. Develop WGS harmonized protocols at each of the steps of
the process
• 2. Backwards comparability between WGS and PFGE :
Set up of a EU wide relevant strain/ genome collection
Set a cut-off for WGS among the genetic diversity of L.
monocytogenes
Cluster definition : assess the genetic distance between
epidemiologically related / not-epidemiologically related
strains.
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COllaborative Management Platform for detection and Analyses of
(Re-)emerging and foodborne outbreaks in Europe
-Coordinator : DTU-Food, (Franck Aarestrup’s team)
-29 European partners ; 15 Work packages
-Anses : Maisons-alfort laboratory for food safety ; Anses WGS
platform (Ploufragan, Brittany).
2) The EURL project on WGS typing
(2015-2017)
In close collaboration with :
The EURL/NRL (in particular IT, DE) network and other Anses
centers
1) The European project
(2015-2020)
EURL for Lm
European Union Reference
Laboratory for
Listeria monocytogenes
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Assess the typing potential of WGS /research
projects
Overview of the genetic structure of populations of food and
clinical strains ;
Explore the genetic diversity of strain populations in France and
in Europe
Assess the epidemiological relationship of strains from the
different sources
Identify the presence of molecular markers that may explain
the virulence / ecophysiology differences observed within the
strain populations.
PhD project Anses /DTU-food (2013-2016)
2 years-European project (2014-2016)
18 month project -National Grant) (2015-2017)
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14
WGS : a maximal resolution typing tool for molecular
surveillance and outbreak investigation ?
WGS : Information on
pathogen genomes (species
identification, drug
resistance, virulence,
evolution…).
MLST : Global epidemiology
Population biology
Phylogeny
SérotypeLineage MLST PFGE WGS
Discriminatory degree
PFGE : The current international
standard -Outbreak
investigation
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WGS : a roadmap to better manage the food safety…
Research projects
Molecular markers underlying
evolution, host specificity,
virulence, transmissioon
ecophysiology…
1. Stimulate all the
partner networks to
track and type more
strains (+ historical
strains)
3.Harmonization within
clinical and food reference
lab networks
4. Share relevant
information/
databases
Networking
Competent authorithies
: facilitate
communication and data
sharing between all the
actors
2. Design a
rational typing
scheme for
outbreak
investigation
type of WGS
data useful for
surveillance ?
Define
standardized
nomenclature
Validation by
reference
laboratories
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A standardized (cgMLST) : intermediate between
SNP-based phylogeny and MLST ?
Molecular
surveillance
Molecular
methods
Genotyping –based
surveillancePFGE
SNPs
MLST
allele-based typing scheme
Wg MLST –cgMLST (more
discriniating than
MLST)(Ruppitsch et al., 2015) ;
Jolley; Miaden, 2014);
Genomic-based
surveillance
Nomenclature standardized
Central database; not enough
discriminating
interpretation of data easier
and more accessible than
PFGE
Central databases (ex.
BIGSDB)
highly discriminatory, less
standardized
No standardized
nomenclature and profile
interpretation difficult
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Perspectives : WGS in France /Europe– replacement of PFGE ?
• Easier to replace PFGE with WGS at the local level than in large
national / international surveillance networks.
• The replacement of PFGE with a new WGS-based method must be
conducted gradually to avoid the loss of precious historic
information generated over many years.
• Underscored by the continued use of PFGE.
• In French lab/ European countries with limited financial ressources,
PFGE will probably prevail in the coming years
• But these labs may rapidly adopt WGS once it is practical to use
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Acknowledgement
SEL Unit
Listeria team - molecular typing team
Clémentine Henri
Trinh-Tam Dao
Karol Romero
Jean-François Mariet
Damien Michelon
Benjamin Félix
Renaud Lailler , david Albert
Maisons-Alfort Food Safety
laboratory
Michel-Yves Mistou
Laurent Guillier
Nicolas Randomski
Arnaud Felten
Olivier Firmesse
Anses WGS Platform
Yannick Blanchard
EURL Lm coordination
Bertrand Lombard
Adrien Asséré ; Marie-Christine Salvi
Food NRL network, in particular (AT, BE, DE,
FI, IE, IT, NL, SE)
Vicdalia Aniela Acciari
Nadine Botteldoorn
Els Biesta-Peters
Bernadette Hickey
Luigi Iannetti
DTU-food :
FM Aarestrup
R. Hendriksen
SSI
Eva Moller Nielsen
Jonas Larsson
CDC
Peter Gerner-Smidt
Applied Math’s team
Sylvia Kleta
Ariane Pietzka
Leila Rantala
Susanne Thisted Lambertz
Caroliene van Heerwaarden