Enteric caliciviruses were detected in humans and animals incoming into food chain such as porcine and bovine species. Caliciviruses have a single stranded, positive, polyadenylated RNA genome. They share properties of high environmental stability, high excretion load and high genetic variation linked to point mutations and recombination. These properties have allowed the formulation of hypothesis about zoonotic transmission or animal reservoir for human strains. The aim of the thesis, composed of four studies, was to investigate the zoonotic risk associated to animal enteric caliciviruses.
In a first study, circulation of both sapoviruses and noroviruses was evidenced by molecular detection in Belgian pig farms. They were detected in both asymptomatic animals or in piglets showing clinical signs of enteritis.
In a second study, bovine noroviruses were molecularly detected in Belgian cattle. Strains phylogenetically related to those of the genotype 2 were predominant. In the same study, seroprevalence against bovine norovirus infection in cattle was investigated by indirect ELISA. Antigens included in the ELISA were virus-like particules obtained in the baculovirus system by expressing the capsid protein of a strain isolated by the laboratory during a previous study. Apparent seroprevalence was high (93.2%), confirming previous results about apparent molecular prevalence in diarrheic calves (7.5%).
In a third study about molecular detection of bovine noroviruses, diagnostic strategy was revised in order to improve the detection of genotype 1 strains and to deal with opportunity of recombination events. Bovine norovirus recombinant strains and also, surprisingly, some sequences genetically related to bovine kobuviruses were detected.
In a fourth study, attachment factors and internalization pathways for genotype 2 bovine noroviruses were studied with an original quantitative method based on flow cytometry analysis. Along with a galactosyl residue that seems to be essential, a sialic acid residue was also showed to be implied in the binding of genotype 2 bovine noroviruses or in a posterior step. Internalisation pathways related to lipid rafts and to macropinocytosis were found.
Together the results have contributed to the analysis of the zoonotic risk associated to enteric animal caliciviruses in the Belgian epidemiological situation. According to these results, the zoonotic risk seems to be low as no sequences genetically related to the human ones were detected. However, some results suggest to maintain a certain degree of vigilance. Indeed, molecular detection showed the co circulation of both bovine and porcine noroviruses in Belgian farms, implicating that hazard exposition exists and could be high in the Belgian epidemiological situation. Morevover, circulation of recombinant strains in the overall population of the bovine norovirus strains implies that this phenomenon was included in the risk assessment. Infection pressures and high prevalences for human and animal strains in a closed epidemiological context as the Belgian one could increase the risk of interspecific recombination. Finally, the sialic acid residue, possibly involved in the binding of genotype 2 bovine noroviruses, and a poorly specific internalisation pathway as macropinocytosis could also favor interspecies barrier crossing.
In the current knowledge, zoonotic risk associated to animal enteric caliciviruses can be communicated as low but a degree of vigilance has to be retained, associated for example to observation of genetic evolution in the populations of human and animal strains.