Cell-culture bases techniques for the determination of the infective status of waterborne PATHOgenS

SCHEME: CORE

CALL: 2008

DOMAIN: SR - Sustainable Management of Water Resources

FIRST NAME: Henry-Michel

LAST NAME: Cauchie

INDUSTRY PARTNERSHIP / PPP: No

INDUSTRY / PPP PARTNER:

HOST INSTITUTION: LIST

KEYWORDS: Waterborne diseases

START: 2009-05-01

END: 2012-04-30

WEBSITE: https://www.list.lu/

Submitted Abstract

Safe water is a mandatory resource for human development. Water is indeed a limited resourcethat has no substitute. Its management is therefore of prime importance. Together with somesynthetic chemical compounds, viral and parasitic pathogens are emerging as the majorchallenges of the beginning of the 21st century. Beside lacks of knowledge concerning thesepathogens (survival in the water distribution system, genetic drift and evolution, consumersusceptibility,…), there is a lack of sensitive methods to detect them in water and theassociated biofilms. The Department “EVA” of the Centre de Recherche Public – GabrielLippmann (CRP-GL) has participated to some extent during these last years to themethodological and intellectual advances on this topic. On the basis of the current state of theart, it appears that the next step to be taken to have efficient tools for a correct risk assessmentof the non-bacterial risk in water is the development and validation of methods for thedetermination of the infective status of the pathogens in water. The present project proposes todevelop such methods for two major emerging waterborne pathogens: the protozoan parasiteCryptosporidium parvum and noroviruses. These pathogens have been targeted becauseprevious studies in the CRP-GL have highlighted their presence in the surface waters andgroundwater used for drinking water production in Luxembourg. Top ranking of thesepathogens has also been made by the COST action 929 “European Network for Environmentaland Food Virology” and the FBVE (Foodborne Viruses in Europe) network. Contacts with thesegroups helped us in defining the most relevant actions to be taken in the present project.The main aim of the present project is to develop an integrated detection technique thatcombines cell culture and a highly sensitive molecular tool (real-time immuno-PCR) forthe determination of the infective status of Cryptosporidium parvum and noroviruses. Afirst work package will deal with the establishment of validated cell culture protocols. The mainchallenge is to use a three-dimensional cell culture system for the cultivation of noroviruses, asmonolayer cell cultures have proved to be inefficient in culturing these viruses. A second workpackage will concern the development of protocols for the detection of Cryptosporidium andnoroviruses using real-time immuno-PCR. A third work package will aim the integration of cellculture and real-time immuno-PCR detection using purified pathogens as targets. In a fourthwork package, the integrated cell culture – real-time immuno-PCR protocols validated in workpackage 3 will be applied to environmental samples (surface waters used for drinking waterproduction or recreation activities, groundwater used for drinking water production and thedrinking water during treatment processes and in the distribution systems. Major expectedresults are the establishment of an efficient and reliable technique for detecting the infectiveCryptosporidium and noroviruses in environmental samples and the integration of thisinformation in the risk assessment of waterborne diseases. Results will be disseminatedthrough scientific publications, public information during fairs, oral or poster presentations atinternational conferences and the organisation in Luxembourg of the 2nd European Symposiumon Waterborne Pathogens.

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