integrated monitoring and control of foodborne viruses in european food supply chains vital wim h....
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Integrated monitoring and control of foodborne
viruses in European food supply chains
VITAL
Wim H. M. van der Poel
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Contamination routes of foodborne viruses of
human and animal origin Animals
Animal produce
i.e. Milk, eggs
Faeces / Manure *
Surface water *Crops *
Effluent *
Slaughter *
Meat *
Consumer
Humans *
* VITAL Sampling
points
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Work package 2
Data gathering production phase
Objectives
1. Data gathering viruses on soft fruit and vegetable producing farm: NoV, HAV
2. Data gathering viruses in pig farms and slaughter houses: HEV
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VITAL workpackage 2
human transmission pathway
Sampling points:
1. Field latrines
2. Harvesters’ hands
3. Irrigation waters
4. Soft fruit and vegetables
Exact sampling points to be determined during audit visits
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VITAL workpackage 2
animal transmission pathway
Sampling points:
1. Pig faeces
2. Blood
3. Liver (meat)
4. (Slaughter house effluent)
Exact sampling points to be determined during audit visits
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VITAL workpackage 2
Sampling pig farms and pig slaughter houses
VRI, ISS, ITACyl, (CVI) Pig farms
fecal samples* Pig slaughter houses
Blood Liver (effluent)
*Sampling of feaces transferred to slaughter houses
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VITAL workpackage 2
Sampling salad vegetable farms
DUT, NVRI, NIV-NS Irrigation waters (Animal based fertiliser ?) Field latrines Harvesters’ hands (Vegetable crop ?)
Exact sampling points to be determined during audit visits
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VITAL workpackage 2
Sampling soft fruit farms
UH, VRI, NVRI, NIV-NS Irrigation waters (Animal based fertiliser ?) Field latrines Harvesters’ hands (Fruit crop ?)
Exact sampling points to be determined during audit visits
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VITAL workpackage 2
Testing of samples
Testing of collected samples by data gathering laboratories
Forwarding data for analyses
VRI, ISS, ITACyl, CVI, NVRI, NIV-NS, UH, VRI
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WP 2
Task 2.1 Preparatory activities
Realtime RT-PCR methods NoV HAV HEV AdV, BPV
Virus sampling /concentration 9 different matrices
NA extr. methods 4 different methods
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WP 2
Task 2.1 Preparatory activities (ctd) Finalising of, and Agreement on SOPS
Virus sampling+concentration protocols rRT-PCR testing protocols
Acquisition of control materials IAC (partner 12 efforts) External proces control (selection by p8) Neg. extraction controls Positive controls (target DNA, target virus) (culture by
p6)
norovirus
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WP 2
Task 2.1 Preparatory activities (ctd)
Training of testing procedures Intra-laboratory training of SOPs Pre-collaborative ring trials
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University of Barcelona Bovine Adenovirus rRT-PCR protocol Bovine Polyomavirus rRT-PCR protocol
University of Washington Murine NoV
Signatures from beneficiaries completed Dec2008
WP2
Task 2.1 Preparatory activities (ctd)
Material transfer agreements (MTAs)
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WP 2, Task 2.1 Preparatory activities (ctd)SOPs for sampling and virus concentration
Number SOP
001 faeces
002 harvesters’ hands
003 animal-derived fertilizer
004 waters
005 soft fruit
006 vegetables
007 shellfish
008 blood
009 liver tissue and pork meat
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WP 2, Task 2.1 Preparatory activities (ctd)SOPs for Nucleic Acid extraction
Number SOP
010 faeces, animal derived fertilizer, or blood
011 pork liver tissue or meat
012 soft fruits, vegetables or shellfish
013irrigation water, slaughterhouse effluents, or harvesters’ hands wash-off
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WP 2, Task 2.1 Preparatory activities (ctd)SOPs for QPCR testing
Number QPCR SOPs
014 General adenovirus QPCR (From VIROBATHE project )
015 Detection and quantification of porcine adenoviruses (UB)
016 Bovine polyomavirus QPCR
017 Human adenovirus nested PCR
018 Norovirus ggII
019 Norovirus ggI
020 HAV QPCR
021 HEV QPCR
022 Standard Proces Control (Murine NoV) QPCR
023 Porcine adenovirus IAC
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WP 2, Task 2.1 Preparatory activities (ctd)
Pre-collaborative ring-trial
Target virus (spiking virus) Human Adenovirus Standard proces control: Murine Norovirus Organisation by partner 1, CSL Viruses cultured and shipped by partner 6, ISS Participating labs: all data gathering labs and
T6.3 labs March 2009 – Dec 2009
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VITAL WP2
Data Gathering production phase
Wim H. M. van der Poel
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VITAL workpackage 2
Milestones
M2.1 All data-gathering labs fully prepared with necessary materials and SOPs (M3) Method development and validation extended to March
2009 (M11) M2.2 All data gathered from the production phase
of three food supply chains (M15) Sampling immediately after audit visits (M12- ) Analyses susequently after sampling (M12- )
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VITAL workpackage 2
Deliverables
D 2.1 Data from avenues of virus contamination of salad vegetable crops (M15)
D2.2 Data from avenues of virus contamination of soft fruit crops (M15)
D2.3 Data on virus contamination of pig feces and unprocessed pig products (M15)
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VITAL WP2
Collected samples at Mid-Term
WP Task Number of samples taken until October 31st , 2009
Present status
2 2.2 Data-gathering: Salad vegetable farms
100 Samples extracted and stored at -80C or -20C as appropriate
2.3 Data-gathering: Soft fruit farms
212
2.5 Data-gathering: Slaughterhouses
46