title: An in vivo system for directed experimental evolution of rabbit haemorrhagic disease virus.
abstract: The calicivirus Rabbit haemorrhagic disease virus (RHDV) is widely used in Australia as a biocontrol agent to manage wild European rabbit (Oryctolagus cuniculus) populations. However, widespread herd immunity limits the effectiveness of the currently used strain, CAPM V-351. To overcome this, we developed an experimental platform for the selection and characterisation of novel RHDV strains. As RHDV does not replicate in cell culture, variant viruses were selected by serially passaging a highly virulent RHDV field isolate in immunologically naïve laboratory rabbits that were passively immunised 18-24 hours post-challenge with a neutralising monoclonal antibody. After seven passages, two amino acid substitutions in the P2 domain of the capsid protein became fixed within the virus population. Furthermore, a synonymous substitution within the coding sequence of the viral polymerase appeared and was also maintained in all subsequent passages. These findings demonstrate proof-of-concept that RHDV evolution can be experimentally manipulated to select for virus variants with altered phenotypes, in this case partial immune escape.
authors.affiliation: Health and Biosecurity, Commonwealth Scientific and Industrial Research Organisation, Canberra, ACT, Australia.|Invasive Animals Cooperative Research Centre, University of Canberra, Canberra, ACT, Australia.
authors.forename: Robyn N
authors.initials: RN
authors.lastname: Hall
authors.affiliation: IZSLER, Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna "Bruno Ubertini", Brescia, Italy.
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Capucci
authors.affiliation: Health and Biosecurity, Commonwealth Scientific and Industrial Research Organisation, Canberra, ACT, Australia.
authors.forename: Markus
authors.initials: M
authors.lastname: Matthaei
authors.affiliation: IZSLER, Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna "Bruno Ubertini", Brescia, Italy.
authors.forename: Simona
authors.initials: S
authors.lastname: Esposito
authors.affiliation: Health and Biosecurity, Commonwealth Scientific and Industrial Research Organisation, Canberra, ACT, Australia.|School of Life and Environmental Sciences, University of Sydney, Sydney, NSW, Australia.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Kerr
authors.affiliation: Health and Biosecurity, Commonwealth Scientific and Industrial Research Organisation, Canberra, ACT, Australia.|Invasive Animals Cooperative Research Centre, University of Canberra, Canberra, ACT, Australia.|Health Research Institute, University of Canberra, Canberra, ACT, Australia.|Institute for Applied Ecology, University of Canberra, Canberra, ACT, Australia.
authors.forename: Michael
authors.initials: M
authors.lastname: Frese
authors.affiliation: Health and Biosecurity, Commonwealth Scientific and Industrial Research Organisation, Canberra, ACT, Australia.|Invasive Animals Cooperative Research Centre, University of Canberra, Canberra, ACT, Australia.|Institute for Applied Ecology, University of Canberra, Canberra, ACT, Australia.
authors.forename: Tanja
authors.initials: T
authors.lastname: Strive
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D061046:Biological Control Agents; D017250:Caliciviridae Infections / Q000821:virology; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology; D019020:Directed Molecular Evolution; D017929:Hemorrhagic Disease Virus, Rabbit / Q000235:genetics* / Q000472:pathogenicity; D011817:Rabbits
chemical_list: D000911:Antibodies, Monoclonal; D061046:Biological Control Agents; D036022:Capsid Proteins