Integrated monitoring and control of foodborne viruses in European food supply chains VITAL Wim H. M. van der Poel

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

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

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

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

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

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

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

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

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

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

WP 2 Task 2.1 Preparatory activities (ctd) Training of testing procedures Intra-laboratory training of SOPs Pre-collaborative ring trials

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)

WP 2, Task 2.1 Preparatory activities (ctd) SOPs for sampling and virus concentration NumberSOP 001faeces 002harvesters’ hands 003animal-derived fertilizer 004waters 005soft fruit 006vegetables 007shellfish 008blood 009liver tissue and pork meat

WP 2, Task 2.1 Preparatory activities (ctd) SOPs for Nucleic Acid extraction NumberSOP 010faeces, animal derived fertilizer, or blood 011pork liver tissue or meat 012soft fruits, vegetables or shellfish 013 irrigation water, slaughterhouse effluents, or harvesters’ hands wash-off

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

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

VITAL WP2 Data Gathering production phase Wim H. M. van der Poel

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- )

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)

VITAL WP2 Collected samples at Mid-Term WPTaskNumber of samples taken until October 31st, 2009 Present status 22.2 Data-gathering: Salad vegetable farms 100Samples extracted and stored at -80C or - 20C as appropriate 2.3 Data-gathering: Soft fruit farms Data-gathering: Slaughterhouses 46

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