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

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

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

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

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

wim.vanderpoel@wur.nl

© Wageningen UR