VITAL: “Integrated Monitoring and Control of

Foodborne Viruses in European Food supply

chains”

Control y análisis de virus contaminantes en la cadena de

producción alimentaria europea

Bilbao, November 2011

Objective: The concept of VITAL is the integrated risk assessment and management of contamination of the European farm to market food supply chain by pathogenic viruses. Specific objectives: 1. To acquire data on virus contamination of food and environmental sources suitable for quantitative viral risk assessment 2. To assess foodborne viral risks for determining high risk situations and efficacy of interventions 3. To develop new measures to prevent virus contamination of foods and the environment 4. To develop and assess measures for virus reduction and control in case of virus contamination

Resultados más destacados: Parte de los resultados están actualmente siendo evaluados, ya que el proyecto ha terminado en septiembre 2011. Se han adaptado los protocolos existentes para la cuantificación de virus en alimentos, agua y superficies y se han producido datos interesantes de presencia de patógenos virales como norovirus y hepatitis E y de la aplicabilidad e interés de la utilización de indicadores virales de contaminación, como los adenovirus humanos y los adenovirus porcinos. También se dispone de datos sobre la eficiencia de tratamientos de desinfección como cloro o radiaciones UV sobre adenovirus. Se están llevando a cabo estudios de evaluación del riesgo con los datos obtenidos en el proyecto.

General conclusion of the project: Overall, VITAL has demonstrated that there are vulnerabilities in the food supply chains, and that these vulnerabilities can occur at critical points of non-compliance with good practice. Future strategies could include adopting controls at these critical points, and evaluating the effect of control measures on virus contamination, by monitoring the indicator viruses after control.

Participantes: El coordinador es Nigel, Cook, Defra, United Kingdom. El total de participantes son 13 laboratorios europeos Más información Página web del proyecto: eurovital.csl.gov.uk Página web del laboratorio de Rosina Girones: www.ub.edu/microbiologia/virology Acciones producidas • Code of Good Practice for Producers to control foodborne viral contamination • Symposium: "New Developments in Monitoring and Control of Viruses in Food

Supply Chains" • Workshop: "Taking Forward New Developments in Monitoring and Control of

Viruses in Food Supply Chains" Training Course for Analysts: "Monitoring of Viruses in Food Supply Chains"

• Training Course for Risk Managers in the Food Industry: "Control of Viruses in Food Supply Chains"

Waterborne

infections

Foodborne

infections

Wastewater

Viral excretion

Food handling

Seawater

Irrigation water

Drinking water

A long trip through the environment

Vital’s

monitoring

methodology

and target

viruses

VITAL: “Integrated Monitoring and Control of Foodborne

Viruses in European Food supply chains”

Work package no. Work package title Type of activity

1 Project management MGT

2 Data-Gathering: Production RTD

3 Data-Gathering: Processing RTD

4 Data-Gathering: Point of sale RTD

5 Data analysis RTD

6 Control measures RTD

7 Delivering impact RTD

MGT: Management of the consortium RTD: Research and technological development.

HAdV have a double role as pathogens/indicators of human

fecal contamination

Adenoviruses have been described as pathogens and as indicators of fecal

contamination as they are excreted by human populations worldwide in high loads and

therefore, they are present in the environment and food.

(Puig et al. 1994, Pina et al. 1998 , Formiga-Cruz et al. 2002, Bofill-Mas et al. 2006)

Human adenoviruses are stable in the environment and highly resistant to treatments

comonly used in wastewater and drinking water treatment plants.

(Meng and Gerba 1996, Gerba et al. 2002, Thompson et al. 2003)

1. Stability of infectious HAdV in water and food

At dark, infective HAdV present in the sources of fecal contamination remain stable

at lower temperatures.

Simulated solar radiation has an effect on HAdV infectivity in all water matrices

showing 1-2,5 logs of reduction when comparing to the same conditions developed

at dark.

At 37°C, a thermal related decay is observed in wastewater, mineral water

containing wastewater (1/1000) and seawater. 6-log reduction that may be due to

biotic factors were found. In these conditions, this decay overlaps the described

effect of the simulated sunlight.

1.1 Stability of infectious HAdV in potential sources of fecal

contamination

Both at dark and under sunlight

simulation, HAdV infectivity has been

observed to remain stable at 4°C in

lettuce and strawberry samples.

At 30ºC, temperature has an important

role in inactivating HAdV in these food

matrices. Almost 4-log reductions have

been detected both at dark and under

sunlight simulation conditions.

Our results suggest temperature as the

main factor inactivating viruses in lettuce

and strawberries.

2.1 Stability of infectious HAdV in food

2. HAdV inactivation by elimination procedures applied in

food industry

Water matrices studied:

Buffered demand free water (BDF)

Natural sea water

Artificial sea water

Experimental conditions: spiked water is treated in

glassbeakers for 60 minutes. The free chlorine dose is

measured during the assay by a colorimetrical method.

Non-chlorinated controls are also developed in parallel.

Sampling: water samples are taken at different times

from 0 to 1 h (0, 10, 20, 30 and 60 min) and HAdV

infectivity and qPCR are immediately analyzed.

2.1 Chemical-based elimination procedures: HAdV chlorine

disinfection in water

2.1 Chemical-based elimination procedures: main conclusions

In water receiving a 10% of sewage, a Ct value of 22 minutes has been calculated

for obtaining a 2-log reduction of HAdV infectivity.

The obtained kynetics of HAdV inactivation fit the Efficiency Hom Model (EFH).

Once the parameters for the model are estimated, the time values for 2, 3 and 4-

log inactivation of the viruses have been also estimated.

In the tested conditions, no significant differences have been observed between

HAdV inactivation in natural seawater and in artificial seawater. These results

suggest that regarding viral disinfection, artificial seawater might be used in

shellfish depurating plants.

2.2 UV-light radiation of HAdV stocks: main conclusions

Disagreggation based on glycine treatments applied to viral stocks show

good results when observing by electron microscopy.

Although qPCR results show a lower decay due to the UV treatment, 1- log

of HAdV inactivation has been detected. This results are consistent

considering that UV radiation has an incidence to viral genomes.

HAdV type 2 show high stability to UV-light irradiation by low pressure

lamps (253,7 nm fluence), achieving around 2 logs of decay after the

treatment.

HAdV presence in mussel samples has been detected both before and after

depuration.

Infectious adenoviruses have been identified by IFA both before and after

depuration in shellfish samples.

The depuration processes analyzed in this study are not totally efficient to

achieve the complete elimination of viral pathogens potentially present in

shellfish samples.

2.3 Inactivation of HAdV in shellfish depurating plants: Main

conclusions

Participating institutes Country

Defra United Kingdom

Catholic University of Leuven Belgium

Veterinary research institute Czech Republic

University of Helsinky Finland

University of Patras Greece

Instituto superiore di sanita Italy

National institut for public health and the environment Netherlands

Wageningen University Research Netherlands

National veterinary research institute Poland

Scientific veterinary institute “Novi Sad” Serbia

University of Ljubljana

Slovenia

Instituto tecnológico Agrario de Castilla y León Spain

Universitat de Barcelona Spain

Vital’s partners

Our team!

http://www.ub.edu/microbiologia/virology/index.html

Aiora Aregita

Anna Carratalà

Ayalkibet Hundesa

Sandra Fresno

Jesús Rodriguez

Marta Rusiñol

Byron Calgua

Laura Guerrero

Rosina Girones

Sílvia Bofill

We thank Regina Sommer from the

Medical University of Vienna for

collaborating in the UV studies. We also

thank Adriana A. Correa, a visiting scientist

from the University of Florianópolis for

collaborating in seawater chlorination

studies.